AIM: To investigate and compare the diagnoses and treatment of premenstrual syndrome (PMS) and premenstrual dysphoric disorder (PMDD) from the perspectives of psychiatrists and obstetricians/gynecologists (OB/GYNs) in Japan. METHODS: Between December 2021 and February 2022, a web-based survey was conducted among the members of the Japanese Association of Neuro-Psychiatric Clinics. Data from 262 psychiatrists who responded to the aforementioned survey were compared with data from 409 OB/GYNs from a survey conducted in 2021 among members of the Japanese Society of Obstetrics and Gynecology. RESULTS: Overall, 79.8% of psychiatrists and 97.3% of OB/GYNs were involved in practicing PMS/PMDD diagnosis and treatment. Most psychiatrists believed that PMS should be treated by OB/GYNs (74.4%) and PMDD by psychiatrists (75.6%). Only vague medical interviews were conducted by 86.6% of psychiatrists, and only 9.7% maintained a two-cycle symptom diary. Psychiatrists mostly prescribed selective serotonin/serotonin and noradrenaline reuptake inhibitor (SSRI/SNRI) continuous dosing (91.1%), followed by Kampo medicines, especially Kamishoyosan (73.3%); only 2.8% chose oral contraceptive pills, unlike OB/GYNs, while SSRI continuous (32.8%) and luteal phase dosing (20.6%) and Kampo medicine (42.1%) were the most common first-line treatments. Lifestyle guidance was prescribed by 63.6% of psychiatrists, followed by cognitive behavioral therapy (13.8%) and the symptom diary observation method (11.1%), which were similar to OB/GYNs' choices. CONCLUSIONS: Many Japanese psychiatrists and OB/GYNs do not base PMS/PMDD diagnoses on prospective monitoring methods using specific diagnostic criteria and therefore do not provide evidence-based treatment. Moreover, a tendency of being biased toward treatments in which the department specialized was observed.

PURPOSE: The validity and reliability of the Japanese version of the Daily Record of Severity of Problems (DRSP-J) have already been confirmed in a population with premenstrual symptoms. This study aimed to assess the validity and reliability of the DRSP-J in the general population. PATIENTS AND METHODS: We analyzed data from 113 Japanese women with regular menstrual cycles who applied to participate in an ongoing study. Participants were recruited regardless of the severity of premenstrual symptoms, and their menstrual cycles were monitored using the DRSP-J for two cycles. Reliability was examined using Cronbach's α, a measure of internal consistency, and test-retest reliability. Structural validity was assessed using a principal component analysis (PCA). The Hospital Anxiety and Depression Scale (HADS) and EuroQol-5 Dimensions-5 Levels (EQ-5D-5L) were used to examine concurrent validity. The agreement between the clinical diagnoses based on the diagnostic criteria for premenstrual syndrome/premenstrual dysphoric disorder and that based on the DRSP-J score was examined using the kappa coefficient. RESULTS: Cronbach's α for DRSP-J total score was 0.96. The DRSP-J total score showed high test-retest reliability. The PCA showed a two-factor model describing "Mood" and "Behavior/Physical" symptoms. The DRSP-J total score was highly correlated with the HADS total and EQ-5D-5L scores. The classification of "moderate to severe premenstrual syndrome" and "premenstrual dysphoric disorder" by clinical methods and the criteria based on the DRSP-J were in good agreement (kappa values = 0.78). CONCLUSION: The DRSP-J is a reliable and valid measure of premenstrual symptoms in the general Japanese population, including those with few or no symptoms.

日本の更年期障害治療における補完代替療法,とくにイソフラヴォン類とプラセンタ製剤の使用実態を明らかにする目的で,以下の検討を行った.日本産科婦人科学会女性ヘルスケア委員会では,日本の更年期障害治療の実態を把握するために,日本女性医学学会員の医師全員を対象として,2020年に質問紙郵送による調査を行った.本調査についてはこれまで全体の概要,および漢方薬の使用実態についての報告を行ったが,今回は補完代替療法,とくにイソフラヴォン類とプラセンタ製剤に着目した.調査票上ではイソフラヴォン類としてエクエル,プラセンタ製剤としてメルスモン・ラエンネックを列記し,その他は自由記載とした.得られた回答に基づいて各治療法の(1)現在使用率=現在使用している医師数/全医師数(%),(2)有効率=有効と考える医師数/現在・過去使用医師数(%),(3)有害事象経験率=有害事象経験医師数/現在・過去使用医師数(%)を算出した.1,532名より回答を得た(回答率43.5%).全体の現在使用率は,エクエル51.4%,メルスモン17.5%,ラエンネック7.2%で,エストロゲン製剤(41～86%)より低かった.有効率は,エクエル71.2%,メルスモン76.5%,ラエンネック70.9%で,エストロゲン製剤(89～95%)より低かった.有害事象経験率は,エクエル3.2%,メルスモン4.8%,ラエンネック1.2%で,エストロゲン製剤(6～17%)より低かった.月あたりの更年期障害患者数が増えるほど,エクエルとメルスモンの現在使用率と有効率が有意に増加した.イソフラヴォン類とプラセンタ製剤の現在使用率・有効率・有害事象経験率はエストロゲン製剤よりも低かった.患者数が増えるほど,これらの製剤を有効に投与できるようになる傾向が見られた.(著者抄録)

AIM: To investigate the current status and problems in the diagnosis and treatment of premenstrual syndrome (PMS) and premenstrual dysphoric disorder (PMDD) from the perspective of obstetricians and gynecologists (OB/GYNs) in Japan, the Japanese Society of Obstetrics and Gynecology (JSOG) conducted a national-wide survey. METHODS: An email survey was sent to all JSOG members (16 732) and a web-based survey was conducted using a Google form between September and November 2021. The current status and problems in PMS/PMDD diagnosis and treatment were surveyed in this cross-sectional study. RESULTS: In total, 1312 respondents (7.8% of all JSOG members) completed the questionnaire. In terms of diagnoses and treatment, OB/GYN was preferred over psychiatrist for PMS (91.4% vs. 45%); however, no differences were noted for PMDD (76.1% vs. 73.7%). A total of 1267 (96.6%) respondents engaged in routine PMS/PMDD treatment. Regarding the general diagnosis procedure, 84.4% respondents answered "only a vague medical interview," 8.4% kept a two-cycle symptom diary, and 10.3% used a screening questionnaire. The most commonly used medication was oral contraceptive pills (OCPs) (98.1%), followed by the Kampo, traditional Japanese herbal medicines, Kamishoyosan (73.6%). Concerning first-line drugs for treatment, OCPs were the most common (76.8%), followed by Kampo medicine (19.5%); selective serotonin reuptake inhibitors (SSRIs) were less frequently used (2.6%). Regarding first-line drugs among OCPs, 65.1% respondents reported drospirenone-ethinylestradriol use. CONCLUSIONS: This study indicates that only a few OB/GYNs practicing PMS/PMDD in Japan use a prospective diary, which is an essential diagnostic criterion for PMS/PMDD. Regarding treatment, SSRIs were used less frequently.

PURPOSE: Adolescence is a period of transition from childhood to adulthood where people are vulnerable to stress. The COVID-19 pandemic continues to cause sustained stress in the population. Since the COVID-19 pandemic, social isolation and loneliness have increased. Loneliness is associated with increased stress, psychological distress, and a higher risk of mental illnesses, such as depression. This study examined the association between loneliness, premenstrual symptoms, and other factors in the era of the COVID-19 pandemic among adolescent females in Japan. PATIENTS AND METHODS: A school-based cross-sectional survey of 1450 adolescent female students in Japan was conducted in mid-December of 2021. Specifically, paper-based questionnaires were distributed in class, and the responses were collected. The Premenstrual Symptoms Questionnaire (PSQ), 6-item Kessler Psychological Distress Scale, 3-item Revised UCLA Loneliness Scale (R-UCLA), and Fear of COVID-19 Scale were used as measurement tools. The prevalence of loneliness was defined as a total R-UCLA score ≥ 6. RESULTS: The prevalence of loneliness was 29.0%. The prevalence of serious psychological distress was also high (8.2%), especially in the lonely group (16.0%). Multivariable regression analysis identified the following factors associated with loneliness: second year (odds ratio [OR] 1.53; 95% confidence interval [CI] 1.09-2.14), longer internet use (OR, 1.11; 95% CI, 1.02-1.20), total PSQ score (OR 1.08; 95% CI 1.06-1.11), and psychological distress (OR 1.05; 95% CI 1.01-1.08). CONCLUSION: Adolescent females in Japan showed a high prevalence of loneliness. School year (2nd year), longer periods of internet use, premenstrual symptom severity, and psychological distress were independently associated with loneliness. For clinicians and school health professionals, special concern should be given to the psychological health of adolescent females during the COVID-19 pandemic.

Recently, the term premenstrual disorders (PMDs), which includes premenstrual syndrome and premenstrual dysphoric disorder as a continuum, has been proposed. Although the precise etiology of PMDs remains unknown, the involvement of hormonal fluctuations is clear. The brain transmitters, serotonin and γ-amino butyric acid, also seem to be involved. Serotonin reuptake inhibitors and oral contraceptives are the current mainstay of treatment, but these are insufficient. Even the currently used prospective two-period symptom diary is not widely used in actual clinical practice, creating a major problem of discrepancy between research and clinical practice. In this review, I would like to outline the latest information and problems in the etiology, diagnosis, and treatment of PMDs, with an emphasis on promising new therapies.

更年期障害の治療法並びにその有効性・合併症の実態を把握するために、日本女性医学学会員の医師全員(3,520名)を対象として、2020年1〜3月に質問紙郵送による調査を行った。各種治療法につき、(1)現在使用率=現在使用している医師数/全医師数(%)、(2)有効率=有効と考える医師数/現在・過去使用医師数(%)、(3)有害事象経験率=有害事象経験医師数/現在・過去使用医師数(%)を算出した。漢方薬としては当帰芍薬散(TJ-23、以下23)・加味逍遙散(TJ-24、以下24)・桂枝茯苓丸(TJ-25、以下25)・抑肝散(TJ-54、以下54)・補中益気湯(TJ-41、以下41)の5処方を列記し、その他は自由記載とした。1,532名より回答を得た(回答率43.5%)。5処方の現在使用率は、(23)87.6%、(24)92.2%、(25)86.2%、(54)48.4%、(41)49.5%であり、各種エストロゲン製剤よりやや高い傾向にあった。有効率は、(23)85.4%、(24)88.1%、(25)87.0%、(54)87.7%、(41)81.9%であり、各種エストロゲン製剤よりもやや低い傾向にあった。有害事象経験率は、(23)3.1%、(24)4.0%、(25)2.8%、(54)2.2%、(41)1.3%であり、各種エストロゲン製剤よりも低い傾向にあった。自由記載欄には頻度が高い順に半夏厚朴湯、女神散、桃核承気湯、加味帰脾湯、柴胡加竜骨牡蛎湯などがあげられた。また月あたりの更年期障害患者数が増えるほど、すべての処方の現在使用率と有効率が増加した。(著者抄録)

AIM: Premenstrual syndrome causes disturbances in many women's daily activities. Isoflavones might cause changes in the estrogen cycle by their selective estrogen receptor modulator-like activities. Equol, which is a metabolite of a soy isoflavone, has greater biological activity than other soy isoflavones. In this preliminary study, we aimed to examine the effect of a natural S-equol supplement (SE5-OH) on premenstrual symptoms. The gut microbiota has recently been suggested to play an important role in brain function in psychiatric disease, such as depression. Therefore, we further aimed to evaluate the relationship of the effect of SE5-OH and the gut microbiota at preintervention. METHODS: Twenty women who showed premenstrual symptoms and were nonequol producers were enrolled in an open-label, single-arm, clinical study in which they received oral SE5-OH for two period cycles. The Daily Record of Severity of Problems (DRSP) total score was evaluated during the intervention cycles. Before taking SE5-OH, fecal samples were obtained and subjected to terminal restriction fragment length polymorphism analysis. RESULTS: The response rate to treatment (≥50% reduction from baseline in the DRSP total score) was 10.5%. Post hoc analysis showed a significant improvement in the change in the DRSP total score (P = .008) and DRSP scores for four core premenstrual dysphoric disorder symptoms. Multiple regression analysis showed that the percentage improvement of the DRSP total score was positively related to Bifidobacterium and negatively related to Clostridium cluster IV. CONCLUSION: SE5-OH supplementation may be an acceptable treatment for premenstrual symptoms. The intestinal microbiota may have an effect on SE5-OH.

Purpose: The Premenstrual Symptoms Questionnaire (PSQ) is a patient-reported outcome measurement tool for premenstrual symptoms. Although the PSQ is a very useful tool with 14 items to screen for a wide variety of premenstrual symptoms, not everyone will respond to all the questions. Fewer questions would be less burdensome on the respondents. We aimed to develop and analyze the psychometric properties of a short-form of the PSQ (PSQ-S). Patients and Methods: The study participants were from an earlier study with a sample of 1388 female students. We reanalyzed data collected from 922 students with regular menstrual cycles who completed the PSQ, premenstrual dysphoric disorder (PMDD) scale, Somatic Symptom Scale-8 (SSS-8), and numerical rating scale (NRS) for menstrual pain. First, we selected nine items for the PSQ-S based on the results of the corrected item-total correlation analysis. The PSQ-S was then analyzed for reliability and validity using the classical test theory. Moreover, item response theory was applied to test the psychometric properties of the PSQ-S. Results: Cronbach's α for the PSQ-S score was 0.93. Principal component factor analysis revealed a one-factor model. Confirmatory factor analysis showed that the one-factor model was a good fit. The PSQ-S total score was strongly correlated with the PSQ total score, PMDD scale score, and SSS-8 score (r=0.978, 0.854, and 0.648, respectively) and moderately correlated with the NRS (r=0.437). Item response theory analyses showed that the constructs and items of the PSQ-S had satisfactory discrimination, difficulty parameters, item information curves, and test information curves. Receiver operating characteristic curve analysis revealed a cut-off score of ≥22 for suspected premenstrual disorders based on the total PSQ-S score. Conclusion: The PSQ-S, consisting of nine items from the PSQ, had sufficient reliability and validity and could be a convenient assessment tool for premenstrual symptoms in routine clinical practice.

The relationship between fish eating habits and menstrual pain is unknown. Elucidating this relationship can inform dietary guidance for reproductive age women with menstrual pain. The aim of this study was to clarify the relationship between fish intake frequency/preference and menstrual pain. This cross-sectional study was conducted at the Miyagi Regional Center as an adjunct study of the Japan Environment and Children's Study, and 2060 eligible women (mean age, 31.9 years) participated. Fish intake frequency ("< 1 time/week," "1 time/week," "2-3 times/week," or "≥ 4 times/week"), preference ("like," "neutral," or "dislike"), and menstrual pain (no/mild or moderate-to-severe) were assessed at 1.5 years after the last delivery through self-administered questionnaires. The association between fish intake frequency/preference and prevalence of moderate-to-severe menstrual pain was evaluated through logistic regression analyses. Our results show that, compared with the "< 1 time/week" (38.0%) group, the "1 time/week" (26.9%), "2-3 times/week" (27.8%), and "≥ 4 times/week" (23.9%) groups showed a lower prevalence of moderate-to-severe menstrual pain (p < 0.01). The prevalence of moderate-to-severe menstrual pain was 27.7%, 27.6%, and 34.4% in the "like," "neutral," and "dislike" groups, respectively. Multivariate logistic regression showed that frequent fish intake was associated with a lower prevalence of moderate-to-severe menstrual pain ("1 time/week": odds ratio [OR] = 0.59; 95% confidence interval [CI], 0.41-0.86, "2-3 times/week": OR = 0.64; 95% CI, 0.45-0.90 and "≥ 4 times/week": OR = 0.52; 95% CI, 0.34-0.80; trend p = 0.004). Multivariate logistic regression showed no association between fish preference and moderate-to-severe menstrual pain ("dislike" vs "like": OR = 1.16; 95% CI, 0.78-1.73). There was a significant negative association between fish intake frequency and menstrual pain. It is suggested that fish intake can reduce or prevent menstrual pain.

PURPOSE: Premenstrual symptoms can negatively impact the quality of life of women through a range of mood, behavioral, and physical symptoms. The association between the microbiota and brain function has been extensively studied. Here, we examined the characteristics of the microbiota in women with premenstrual disorders (PMDs) and the association between premenstrual symptoms and the microbiota. MATERIALS AND METHODS: In this single center cross-sectional pilot study, we recruited 27 women reporting premenstrual symptoms and 29 women with no serious premenstrual symptoms. Among them, we further selected 21 women experiencing premenstrual symptoms resulting in interference to their social life (PMDs group) and 22 women with no serious premenstrual symptoms and thereby no interference to their social life (control group). The severity of symptoms was evaluated by a premenstrual symptoms questionnaire (PSQ). Inflammatory markers were analyzed in blood samples, including C reactive protein, soluble CD14, and lipopolysaccharide binding protein. Sequencing of 16S ribosomal ribonucleic acid genes was performed on stool samples. RESULTS: Inflammatory markers in blood samples did not differ significantly between the PMDs and control groups. A difference in beta, but not alpha diversity, was detected for the gut microbiotas of the PMDs and control groups. The relative abundance of the Bacteroidetes phylum was lower in the PMDs group. At the genus level, the prevalence was decreased for Butyricicoccus, Extibacter, Megasphaera, and Parabacteroides and increased for Anaerotaenia in the PMDs group, but after false discovery rate correction, these differences were no longer significant. Linear discriminant effect size analysis revealed a decrease in Extibacter, Butyricicoccus, Megasphaera, and Parabacteroides and an increase in Anaerotaenia in the PMDs group. The PSQ total score correlated with Anaerotaenia, Extibacter, and Parabacteroides. Multiple regression analysis showed that Parabacteroides and Megasphaera negatively predicted the PSQ total score. CONCLUSION: The properties of the gut microbiota are associated with premenstrual symptoms.

OBJECTIVE: To investigate the interval from menarche to the onset of premenstrual symptoms and its relationship with menarche age. DESIGN: Cross-sectional school-based survey. SETTING: Urban areas of Sendai, the largest city in northeastern Japan. PARTICIPANTS: 1422 female Japanese 10th-12th grade senior high school students participated in the survey. MAIN OUTCOME MEASURES: The time of awareness of premenstrual symptoms, and the interval from menarche to the onset of premenstrual symptoms. RESULTS: 1290 students had menstruation and completed the whole survey. The median age at menarche was 12 years (IQR: 11-13 years). The prevalence of self-reported premenstrual symptoms was 49%. The median age at which students became aware of premenstrual symptoms was 15 years (IQR: 14-16 years). The median time from the onset of menarche to awareness of premenstrual symptoms was 2 years. This time was negatively correlated with menarche age (ρ=-0.47, p<0.001). A Cox proportional hazards regression analysis revealed that early menarche was significantly associated with a lower cumulative risk of developing premenstrual symptoms (OR: 0.73 (95% CI 0.58 to 0.91)). CONCLUSIONS: High school students in Japan began experiencing premenstrual symptoms at around 15 years old, and within 2 years of menarche. This study suggested that social factors other than hormonal factors, such as early menarche, might be associated with the onset of premenstrual symptoms.

BACKGROUND: A correlation between gonadal steroids and depressive symptoms during the perinatal period has long been suggested; however, the underlying mechanism for this relationship remains unclear. METHODS: This study was designed to examine the correlation between gonadal steroid concentrations of umbilical cord blood and postpartum depressive symptoms as well as longitudinal alterations in maternal plasma gonadal steroid concentrations among 204 perinatal women. The levels of postpartum depressive state at 1 month postpartum were evaluated using the Edinburgh Postnatal Depression Scale. RESULTS: Umbilical progesterone, estradiol, and testosterone levels were significantly higher in infants delivered by depressed mothers (870.7 ± 281.7 ng/ml, 8607.7 ± 4354.6 pg/ml, and 2.5 ± 0.9 ng/ml, respectively) than those delivered by nondepressed mothers (741.3 ± 324.0 ng/ml, 5221.9 ± 3416.3 pg/ml, and 2.1 ± 0.6 ng/ml, p < .01, p < .05, and p < .05, respectively). Postpartum plasma progesterone levels of depressed mothers (3.5 ± 3.1 ng/ml) measured in the early postpartum period were significantly lower than those of nondepressed mothers (9.1 ± 9.7 ng/ml, p < .01). The decrease in progesterone from mid-pregnancy to the early postpartum period was significantly higher in depressed mothers than in nondepressed mothers. Subgroup analyses specific to primiparas or multiparas indicated that a significant drop of progesterone was seen only in primiparas. CONCLUSION: The current study suggests that the delivery of a placenta/fetus with high gonadal steroid production may cause a wider range of fluctuations in maternal plasma gonadal steroid concentrations, which may be concurrent with postpartum depressive symptoms.

Purpose: Pregnant women are vulnerable to stress. The coronavirus disease 2019 (COVID-19) has caused a global pandemic and created significant stress for many people. Social distancing to reduce the spread of COVID-19 has also reduced social interactions, which has increased social isolation and loneliness. Loneliness is thought to increase perceived stress, cause psychological distress, and increase the risk of mental illness, such as depression. This study examined the association between serious psychological distress (SPD) and loneliness during the COVID-19 pandemic in pregnant Japanese women. Patients and Methods: An internet survey of 1022 pregnant women in Japan was conducted between June 1 and July 21, 2021. The 6-item Kessler Psychological Distress Scale, 3-item Revised UCLA Loneliness Scale, and Fear of COVID-19 Scale were used as measurement tools. The prevalence of SPD was defined as a K6 score of ≥13. Results: The prevalence of SPD was 16.5%. Multivariate analysis revealed that the risk factors for SPD were younger age (odds ratio [OR] 1.05; 95% confidence interval [CI] 1.01 to 1.10; p = 0.020), history of abortion or miscarriages (OR 1.56; 95% CI 1.04 to 2.36; p = 0.034), unemployment (OR 1.67; 95% CI 1.14 to 2.45; p = 0.008), fear of COVID-19 (OR, 1.12; 95% CI, 1.08 to 1.17; p < 0.001), and loneliness (OR 1.53; 95% CI 1.38 to 1.70; p < 0.001). Conclusion: Pregnant women in Japan showed a high prevalence of SPD. Younger age, unemployment, history of abortion or miscarriages, fear of COVID-19, and loneliness were independently associated with SPD. Clinicians and health officials should pay particular attention to the psychological health of pregnant women during the COVID-19 pandemic.

COVID-19 has caused an unprecedented global pandemic. Premenstrual symptoms include mood-related, behavioral, and physical symptoms that are limited to the luteal phase of the menstrual cycle. Psychosocial stress is a risk factor for premenstrual symptoms. The aim of this study was to examine the association between premenstrual symptoms and stress caused by COVID-19. We analyzed data from 871 students with regular menstrual cycles who completed the Premenstrual Symptoms Questionnaire (PSQ), Fear of COVID-19 Scale, and Impact of Event Scale-Revised version (IES-R). The total PSQ score was significantly higher in women with COVID-19-induced posttraumatic stress symptoms (PTSS) than in non-PTSS groups. Compared with pre-pandemic data (2019), the total PSQ score did not change in non-PTSS, but increased in PTSS groups. All symptoms were more severe in PTSS groups than in non-PTSS groups. Compared with 2019, PTSS groups had more severe symptoms for all symptoms except 'physical symptoms' and 'decreased social activity', and non-PTSS groups only exhibited improvements in the 'decreased social activity'. Multiple regression analysis revealed that the IES-R score was a significant exacerbation factor of the total PSQ score, along with age and menstrual pain. This study revealed the association between pandemic-associated PTSS and the severity of premenstrual symptoms.

Purpose: The Daily Record of Severity of Problems (DRSP) is commonly used to assess premenstrual syndrome and premenstrual dysphoric disorder throughout the world. The aim of this study was to assess the validity and reliability of the Japanese version of the DRSP (DRSP-J). Materials and Methods: We analyzed 324 women with regular menstrual cycles who completed the DRSP-J and the Premenstrual Symptoms Questionnaire (PSQ). They had all applied to participate in an ongoing study for the treatment of their premenstrual symptoms. The DRSP-J was examined for evidence of reliability and validity. To examine reliability, we assessed Cronbach's α, a measure of internal consistency, and test-retest reliability. We assessed structural validity with principal component factor analysis (PCA) and confirmatory factor analysis (CFA). We used PSQ total score to examine concurrent validity. Results: Cronbach's α for DRSP total score was 0.97. DRSP total score and individual items showed high test-retest reliability. PCA showed a two-factor model describing mood and behavioral and physical symptoms. CFA showed that the two-factor model derived from the PCA was an acceptably good fit. DRSP total score correlated moderately with PSQ total score (r = 0.42). Conclusion: The DRSP-J showed reliable and valid measures of premenstrual symptoms in Japanese women.

Objective: The KOSMOS study, a multicenter, randomized, double-blind, placebo-controlled trial, investigated the effects and safety of kamishoyosan (TJ-24), a traditional Japanese medicine, in the treatment of climacteric disorder. Methods: Japanese women with climacteric disorder were administered a placebo during a 4-week run-in period, after which they were classified as placebo responders (R group) if their score on the modified Questionnaire for the Assessment of Climacteric Symptoms in Japanese Women (m-QACS) with excitability and irritability as the primary outcome improved by ≥ 3 points and as placebo nonresponders (NR group) otherwise. Members of the NR group were randomly allocated to receive either TJ-24 or placebo. After 12 weeks, their m-QACS scores, anxiety and depression, sleep, and overall quality of life (QOL) were compared. Results: The TJ-24 and placebo arms in the NR group included 20 patients each. The change in the m-QACS scores of members of the NR group for excitability and irritability at 12 weeks versus baseline was -3.1 ± 1.7 in the TJ-24 arm, a significant decrease, but compared with -2.7 ± 2.2 in the placebo arm, no significant difference was between two arms. However, the proportion of participants whose score improved by ≥3 points was significantly higher in the TJ-24 arm. In the subgroup analysis of premenopausal women, the changes in the score for excitability and irritability were significantly larger in the TJ-24 arm. The incidence of adverse drug reactions or adverse events did not differ between the two arms, and no serious events were reported. Conclusion: Although no significant difference was identified for the primary outcome, a significantly higher proportion of patients who received TJ-24 displayed improvement. Its high level of safety and effects on excitability and irritability in premenopausal women suggest that TJ-24 may be a useful treatment.

Introduction: Problematic Internet use is a serious health issue for modern adolescents who have developed and matured in a global Internet environment. This study aimed to examine whether Internet use time and Internet addiction beliefs were independently associated with menstrual pain severity and its interference among Japanese adolescents.Methods: We recruited 1166 adolescents at two high schools to complete questionnaires on Internet use and menstrual pain. The explanatory variable was self-reported average Internet use time per day and Internet addiction beliefs ('none' (reference) 'somewhat', 'moderate' and 'severe'). The outcome variables were moderate-to-severe menstrual pain and interference. A multivariable logistic regression model was used to estimate odds ratios (ORs) and 95% confidence intervals (Cls) for moderate-to-severe menstrual pain and interference. Adjustment variables were age, body mass index, belonging to a community or school sports club, sleep duration and Internet addiction beliefs/Internet use time.Results: Internet addiction belief, but not Internet use time, was independently associated with menstrual pain severity and interference to social life in a dose-response manner. ORs (95% Cls) of moderate-to-severe menstrual pain for slight, moderate and severe Internet addition beliefs were 1.43 (1.02-1.99), 1.89 (1.31-2.74) and 1.88 (1.10-3.23), respectively. ORs of moderate-to-severe disability due to menstrual pain for slight, moderate and severe Internet addition beliefs were 1.25 (0.75-1.82), 1.72 (1.01-2.92) and 2.21 (1.11-4.40), respectively, after adjusting for average Internet use time.Conclusion: Internet addiction belief was associated with higher prevalence of moderate-to-severe menstrual pain among adolescents, beyond the variance accounted for by Internet use time.

PURPOSE: Analgesic medication epidemic during pregnancy is an important issue in Western countries. However, no large epidemiological study involving pregnant women with pain and their medication use has been conducted in Japan. This study examined the current situation of medication use for non-cancer pain during the perinatal period in Japan using national cohort data. METHODS: We analyzed 94,649 pregnant women who completed a self-report questionnaire investigating bodily pain and an interview-based medication use survey. Medication use before and during pregnancy and new medication administration/discontinuation during pregnancy were compared between women with and without pain during pregnancy using multivariable covariance analysis. RESULTS: Mild pain was reported by 50.4% of pregnant women in the first trimester (survey 1) and 61.8% in the second/third trimester (survey 2). Moderate-to-severe pain was reported by 15.4% of women in survey 1 and 22.4% in survey 2. In survey 1, 6.2% of women used prescribed analgesics and 1.6% used over-the-counter analgesics. In survey 2, prescribed and over the counter analgesics were used by 12.2% and 0.8% of women, respectively. Other pain-related medications were rarely used (< 1.0%). Pregnant women with moderate-to-severe pain showed a lower proportion of discontinuation of analgesics and a higher proportion of new administration of prescription and transdermal analgesics compared with those without pain. CONCLUSIONS: Although a large proportion of pregnant women experience pain, medication use for pain during pregnancy is low in Japan compared with Western countries (50-60%). Adequate treatment or support may be necessary for pregnant women experiencing pain in Japan.

Premenstrual symptoms are common problems among female high-school and college athletes. This prospective study investigated associations between sleep habits and interference of premenstrual symptoms in athletic performance among Japanese adolescent athletes. A school-based survey on menstruation and school life was conducted among 623 female high-school students in 2015 and 2016, and data from the two surveys were connected. In total, 262 students completed the questionnaire in both years. We recruited 108 of those students who were athletes with regular menstrual cycles. Participants completed a questionnaire about their premenstrual symptoms and lifestyle habits. The prevalence of short sleep duration (<8 h) was high (95.4%). The difference in premenstrual symptom severity in individual students between the first and second years was not significant (p > .05). Premenstrual symptoms were less associated with disturbance in 'athletic performance in training or competition' in the second year than the first. 'Longer sleeping time' in the first year was associated with lower risk of increased interference of premenstrual symptoms in athletic performance in the second year (odds ratio 0.982, 95% confidence interval: 0.970-0.994). Shorter sleep duration may therefore have an effect on premenstrual symptoms' interference with athletic performance among Japanese adolescents.

Purpose: Premenstrual symptoms comprise a wide range of mood, behavioral, and physical symptoms occurring during the luteal phase. Perceived injustice is a belief linked to unfairness (ie, unnecessary suffering caused by illness). This study aimed to assess the validity and reliability of the Premenstrual Symptoms Questionnaire (PSQ), a patient-reported outcome measurement tool, and to examine the association between perceived injustice/perception of menstruation and premenstrual symptoms, as measured by the PSQ. Materials and Methods: Of 1388 female students, we analyzed 879 students with regular menstrual cycles who completed the PSQ, the premenstrual dysphoric disorder (PMDD) scale, the Somatic Symptom Scale-8 (SSS-8), and the Injustice Experience Questionnaire-chronic (IEQ-chr). First, the PSQ was examined for evidence of reliability and validity. Next, we used multiple regression and multivariate logistic regression to investigate the association between perceived injustice and premenstrual symptoms, using PSQ score as both a continuous variable and a dichotomous variable (premenstrual disorders or not). Moreover, the association between PSQ score and perceived menstruation was tested using student's t-test and analysis of variance. Results: In terms of reliability, Cronbach's α for PSQ score was 0.93. To assess structural validity, we used confirmatory factor analysis, which showed that the one-factor model and the two-factor model were a good fit. The PSQ showed good agreement with the PMDD scale. In terms of concurrent validity, PSQ total score correlated strongly with PMDD scale score, SSS-8 score, and IEQ-chr score (r = 0.88, 0.69, 0.57, respectively). IEQ-chr score predicted PSQ score (standardized regression coefficient = 0.53; P < 0.0001) and higher prevalence of premenstrual disorders (odds ratio: 1.15; 95% confidence interval: 1.12-1.19). Negative perception of menstruation was associated with premenstrual symptoms. Conclusion: The PSQ showed sound psychometric properties among the adolescents in our sample. Perceived injustice and negative perception of menstruation were associated with premenstrual symptoms.

Objective: Kampo medicine, a traditional Japanese medicine, is widely used in Japan, especially in the field of menopause medicine. However, few studies have shown evidence-based effects. This study aimed to confirm the effects of kamishoyosan on menopausal symptoms with a randomized, placebo-controlled, double-blind clinical trial. Methods: Subjects were randomly allocated to groups that received either kamishoyosan (n = 101) or a placebo resembling kamishoyosan (n = 104). The primary outcomes were the change in the number of hot flashes, depression scores, improvements of anxiety, quality of life (QOL), and menopausal symptoms before and 4 and 8 weeks after initiation of treatment with the study drug. The secondary outcome was drug safety. Results: After 8 weeks, the number of hot flashes decreased after treatment in both groups, but there was no significant difference between the two groups. The changes in SDS scores showed the same results. Moreover, no significant differences were observed between the two groups in assessments with the STAI, SF-36, and JSOG menopausal index. No serious adverse effect was reported. Conclusions: This first placebo-controlled double-blind randomized trial with kamishoyosan demonstrated that it was safe and had some effects on climacteric symptoms, but not significant compared with placebo. Some problems, such as placebo effects, in the study of Kampo therapy for menopausal symptoms, were revealed. This trial is registered with the trial registration number. UMIN 000006042.

BACKGROUND: Dysmenorrhea influences emotional distress as well as physical suffering in young non-pregnant women. The aim of this study was to assess the potential association between preconception dysmenorrhea and the development of psychological distress during pregnancy. METHODS: This study was a part of the Japan Environment and Children's Study (JECS), a nationwide birth cohort study conducted between 2011 and 2014 in Japan. A total of 87,102 pregnant Japanese women with no psychological distress (Kessler 6-item psychological distress scale [K6] score ≤ 12) in early pregnancy were eligible. Among these, 7626 had mild and 1638 had severe preconception dysmenorrhea. The prevalence and risk of maternal psychological distress (K6 scores ≥ 13) in the second or third trimester were compared among preconception dysmenorrhea severity groups. RESULTS: A higher percentage of women with mild (2.6%) or severe preconception dysmenorrhea (3.6%) suffered psychological distress during pregnancy compared to that in women without dysmenorrhea (2.1%). A multilevel logistic regression model, adjusting for baseline characteristics and the K6 score at enrollment, showed that the severity of dysmenorrhea was associated with psychological distress (mild dysmenorrhea: adjusted odds ratio [aOR], 1.154; 95% confidence interval [95% CI], 0.980-1.359; and severe dysmenorrhea: aOR, 1.457; 95% CI, 1.087-1.951). LIMITATIONS: Information about dysmenorrhea was obtained during early pregnancy. The JECS did not have clear diagnostic criteria for dysmenorrhea. CONCLUSIONS: Preconception dysmenorrhea is associated with an elevated incidence of psychological distress during pregnancy. Additionally, expectant mothers with a history of severe dysmenorrhea symptoms before pregnancy have a higher risk of developing psychological distress.

Background: Menstrual pain causes low quality of life among women of reproductive age, and often interferes with daily activities. Perceived injustice is a cognition linked to adverse symptoms. The aims of this study were to develop a Japanese version of the Injustice Experience Questionnaire-chronic (IEQ-chr-J), and to examine if perceived injustice is associated with pain intensity and impairment from menstruation. Methods: We investigated 130 Japanese women (aged 20-45 years) with menstrual pain in the past 3 months using online self-administered questionnaires. We examined the psychometric properties of the IEQ-chr-J including: structural validity; internal consistency; and test-retest reliability (intra-class correlation coefficients; ICC). Concurrent validity was examined by correlations among the IEQ-chr-J, the Pain Catastrophizing Scale (PCS), the Hospital Anxiety and Depression Scale (HADS), a numerical rating scale (NRS) for maximum/average menstrual pain, and the Brief Pain Inventory (BPI) pain interference domain. We used multivariable regression analysis to investigate the association between perceived injustice and severity of menstrual pain, after excluding 10 hormone drug users. Results: The IEQ-chr-J showed sufficient validity and reliability (Cronbach's α = 0.96, ICC 0.75, [95% confidence interval (CI): 0.61-0.88]. Pearson's correlation coefficients for the IEQ-chr-J, PCS, HADS anxiety, HADS depression, NRS, and BPI pain interference ranged from 0.27-0.65. The IEQ-chr-J was correlated with impairment due to menstrual pain (ICC 0.36, 95% CI: 0.14-0.58), an independent diagnosis of endometriosis, anxiety, and depression, but not with maximum or average pain intensity. Conclusions: The IEQ-chr-J has acceptable psychometric properties, and perceived injustice is associated with impairment from menstrual pain.

Females are well known to suffer disproportionately more than males from stress-related neuropsychiatric disorders, especially during perimenopausal and postmenopausal periods. In addition to a decline in serum estradiol levels, environmental stress and social stress likely contribute to the development of neuropsychiatric symptoms in perimenopausal and postmenopausal women. Kamishoyosan (KSS) is a traditional Japanese Kampo medicine, composed of a specified mixture of 10 crude compounds derived from plant sources, widely used for various neuropsychiatric symptoms in perimenopausal and postmenopausal women. However, the molecular mechanisms underlying KSS-mediated attenuation of neuropsychological symptoms and stress-response behaviors in perimenopausal and postmenopausal women remain unknown. In the present study, we first established a mouse model for postmenopausal depression-like signs using chronic water-immersion and restraint-stressed ovariectomized (OVX) mice to investigate the underlying molecular mechanism of KSS. We found that continuous administration of KSS to these mice normalized the activation of the hypothalamic-pituitary-adrenal (HPA) axis, ameliorated stress-induced depressive behavior, and prevented a decrease of neurogenesis in the hippocampus. As previous studies have implicated dysfunction of the hippocampal 5-HT1A receptor (5-HT1AR) in depressive disorders, we also evaluated the effect of KSS on 5-HT1AR expression and the protein kinase A- (PKA-) cAMP response element-binding- (CREB-) brain-derived neurotrophic factor (BDNF) signaling pathway in the hippocampus in this model. The level of 5-HT1AR in the hippocampus decreased in chronic stress-exposed OVX mice, while KSS treatment normalized the stress-induced decrease in 5-HT1AR expression in the hippocampus of chronic stress-exposed OVX mice. Furthermore, we found that KSS treatment upregulated the expression levels of phosphorylated PKA (p-PKA), phosphorylated CREB (p-CREB), and BDNF in the hippocampus in chronic stress-exposed OVX mice. These results suggest that KSS improves neuropsychiatric symptoms through 5-HT1AR and PKA-CREB-BDNF signaling in the hippocampus in postmenopausal women.

症例は頭痛を主訴とする30代女性。特記すべき既往歴なし。子宮内膜症と月経困難症の治療目的で4年来内服を継続している低用量エストロゲン・プロゲスチン配合薬(low dose estrogen-progestin:LEP)であるドロスピレノン・エチニルエストラジオール配合薬の休薬期間に生じる頭痛が増強し、消炎鎮痛薬に抵抗性となったため、患者本人より当科医師に電話相談があった。休薬期間のみ五苓散(TJ-17)および呉茱萸湯(TJ-31)それぞれ7.5g/日、分3をかかりつけ婦人科主治医に処方してもらうよう指示し、内服を開始したところ、著効した。痛みの評価には、簡易疼痛調査票(Brief Pain Inventory)を用いて、痛みの強さと痛みによる生活障害の程度を評価した。初診時の漢方薬内服前の頭痛の強さは平均5/10、日常生活の全般的活動に対して5/10の支障をきたしており、漢方薬内服1年後の休薬期間における頭痛の強さは平均2/10、日常生活の全般的活動に対しては0/10と頭痛により生活に支障はきたしていなかった。LEPの副作用として、休薬期間のホルモン消退に伴う頭痛は服薬コンプライアンスを低下させるが、本症例では消炎鎮痛剤にてコントロール不良の頭痛に対し、五苓散と呉茱萸湯の合方により水毒症状が改善され、良好な鎮痛が得られた。また、国際的な評価尺度を用いて痛みの強さだけでなく、痛みによる生活障害の程度を評価することは、痛みのある患者に対する漢方薬の治療効果を客観的に評価する意味で有用である。(著者抄録)

Aim: Consumption of soy isoflavones reduces the risk of estrogen-related diseases, such as menopausal symptoms, osteoporosis, and cardiovascular disease. Equol is metabolized from the isoflavone daidzein by intestinal bacteria and has higher bioavailability than other isoflavones. Equol producers are believed to benefit from soybean consumption to a greater extent than non-producers. Recently, we showed that equol non-producers were at significant risk of premenstrual syndrome (PMS). Previously, we reported that PMS is a common menstrual problem in female athletes, and almost half of the studied athletes felt a negative effect of premenstrual symptoms on their athletic performance. This study was conducted to evaluate the relation between PMS and equol production status in Japanese collegiate athletes. Methods: This was a cross-sectional, observational study that included 88 Japanese female collegiate athletes. Equol production status was determined using urine samples collected after a soy challenge test. The subjects also completed a questionnaire about their premenstrual symptoms and their competitive career. Results: The prevalence of equol producers was 29.5% in Japanese collegiate athletes. The athletic performance of 54.5% of athletes was found to suffer in competition or in practice due to premenstrual symptoms. In multivariate analysis, equol non-producers (odds ratio, 3.34 95% confidence interval, 1.03–12.20) and restriction of bodyweight (odds ratio, 4.94 95% confidence interval, 1.47–20.00) were shown to be significant risk factors for poor athletic performance. Conclusion: This study showed a relation between athletic performance and equol production status in Japanese collegiate athletes.

Objectives. This study aimed to evaluate the effect of Royal Jelly (RJ) at a dose of 800 mg/day on menopausal symptoms in healthy Japanese postmenopausal women with placebo-controlled design. Material and Methods. A total of 42 healthy Japanese postmenopausal women have been recruited for this study. The subjects were randomized to oral treatment with either 800 mg of protease-digested lyophilized powder of RJ (enzyme-treated RJ) or placebo (800 mg of dextrin) daily for 12 weeks. The level of menopausal symptoms has been evaluated every 4 weeks, using menopausal symptoms questionnaire of Japanese women. Independent t-test was used to evaluate statistical significance of the treatment effects between the two groups. Results and Conclusion. All of the 42 women have completed the trial. There were significant differences related to the anxiety score (P=0.046) and backache and low back pain score (P=0.040) between 800 mg/day enzyme-treated RJ and placebo-treated groups after 12 weeks of administration, and no significant differences were found between the two groups in 4 weeks after intervention. No side effects were observed in either group. This study demonstrates that enzyme-treated RJ supplementation with doses of 800 mg/day is effective in relieving menopausal symptoms such as anxiety, backache, and low back pain in Japanese postmenopausal women.

Premenstrual syndrome (PMS) is psychosomatic disorder that are limited to the late luteal phase in the menstrual cycle. PMS could impair athletic performance. To investigate associations between proportions of dietary plant and animal protein and PMS-related impairment of athletic performance, we surveyed 135 female athletes aged 18-23 years attending Kindai University. Participants belonged to authorized university clubs, all of which have high rankings in Japanese university sports. Participants completed self-administered questionnaires on diet history, demographics, and PMS-related impairment of athletic performance. Total protein, animal protein, and plant protein intake were examined, and the proportion of dietary plant protein was calculated for each participant. We divided athletes into two groups: those without PMS-related impairment of athletic performance (n = 117) and those with PMS-related performance impairment (n = 18). A t-test was used to compare mean values and multivariable adjusted mean values between groups adjustment variables were energy intake, body mass index, and daily training duration. Total protein intake was not significantly different between the groups. However, athletes whose performance was affected by PMS reported higher intake of animal protein (mean 50.6 g) than athletes whose performance was unaffected by PMS (mean 34.9 g). Plant protein intake was lower among athletes with PMS-related impairment (mean 25.4 g) than among athletes without impairment (mean 26.9 g). The proportion of dietary plant protein was lower among athletes with PMS-related impairment (39.3%) than those without impairment (45.9%). A low proportion of dietary plant protein may cause PMS-related athletic impairment among athletes.

AimConsumption of isoflavones, which are predominantly derived from soybeans, reduces the risk of estrogen-related diseases, such as menopausal symptoms, breast cancer, osteoporosis, and cardiovascular disease. Equol is more bioavailable than other soy isoflavones, and equol producers are believed to benefit to a greater extent. This study was conducted to evaluate the relation between premenstrual syndrome (PMS) and equol-production status in Japanese reproductive-age women. MethodsThis was a cross-sectional, observational study. The study included 144 Japanese women aged 20-45 years. PMS patients (n=46) were recruited at three obstetrics and gynecology clinics. Control group women (n=98) who were not receiving therapy for PMS were recruited from the local area by advertisement. The participants' equol-production status was determined using urine samples collected after a soy challenge test. ResultsThe prevalence of equol producers was 41.8% in the control group and 23.9% in the patient group (P=0.042). Using univariate analysis, significant risk factors for equol non-producers were being a PMS patient and being younger. In multivariate analysis with a step-wise model, being a PMS patient (odds ratio, 2.342; 95% confidence interval, 1.021-5.698) was shown to be a significant risk factor for being an equol non-producer. ConclusionThis study showed a relation between PMS and equol-production status in Japanese women.

Traditional Japanese Kampo medicine has been widely used in clinical practice in Japan. Though it is a compulsory subject in Japanese medical schools, a standard educational program in Kampo medicine does not exist. Tohoku University has incorporated Kampo medicine into clinical education via didactic lectures since 2003; however, student evaluations have been lower for Kampo than for all other clinical specialties. We administered a questionnaire about a Kampo medicine course for fifth-year students from 2009 to 2012 and developed an educational program based on feedback obtained. The questionnaire consisted of nine questions (a clear training plan; opportunities for learning, practice, and patient contact; acquisition of medical knowledge and physical examination; learning professionalism; understanding the specialty; overall assessment) that were rated on a 5-point Liked scale along with open-ended questions about the course's strengths and weaknesses. The students responded to the questionnaire after clinical practice in Kampo medicine and other clinical specialty courses. Scores for Kampo medicine and the average of other clinical specialties were compared. All 389 students who participated in Kampo clinical practice answered the questionnaire. In 2009, scores for Kampo medicine for nine questions were lower than for the average of the other clinical specialties. After curriculum reformation involving hands-on training in 2012, all scores except "opportunities to learn about clinical cases" and "opportunities to practice involvement" were higher than the average of all other clinical specialties. In conclusion, we have successfully developed a Kampo medicine educational program for our university through this survey study.

Study Objective: To determine the specific characteristics of premenstrual syndrome (PMS)/premenstrual dysphoric disorder (PMDD) in Japanese collegiate athletes, with a focus on their fish consumption. Design: Cross-sectional study. Setting: A university in Osaka, the largest city in western Japan. Participants and Interventions: The participants were 312 female collegiate students. The study group was composed of 200 students who were members of sport clubs, and the control (nonathletes) group was composed of 112 members of cultural clubs. Main Outcome Measures: Premenstrual symptoms and social activities. Results: The prevalence of moderate to severe PMS and PMDD in the study group was the same as in nonathletes. The prominent feature of premenstrual symptoms in athletes was that the severities of 'physical symptoms' and 'performance in training or competition' were much greater than those of nonathletes (P = .003 and P = .002, Mann Whitney U test). There was a greater effect of PMS and PMDD on athletes, affecting their physical symptoms and performance compared with nonathletes. In terms of dietary habits, 'fish or dried fish' consumption was associated with a decreased risk of poor performance in athletes (odds ratio, 0.61; 95% confidence interval, 0.40-0.92). Conclusion: The results from this study indicate that fish consumption might be positively associated with the relief of PMS/PMDD-induced athletic disturbance.

Background: Premenstrual disorders such as premenstrual syndrome (PMS) and premenstrual dysphoric disorder (PMDD) interfere with the daily lives of adolescents. The causes of PMS and PMDD are unknown, but lifestyle habits, such as regular exercise and taste preference are known to be associated. This study was conducted to investigate how premenstrual symptoms affect the school life in Japanese high school students and whether there was a risk factor for school absenteeism that is dependent on the types of premenstrual symptoms or lifestyle habits. Methods: A school-based survey was conducted in Sendai, an industrial city in Japan. A total of 901 girls aged 15-19 with regular menstrual cycles were assessed using the self-reporting premenstrual symptoms questionnaire (PSQ) and questions regarding school absence, taste preference, and exercise. We classified the girls into 'no/mild PMS', 'moderate-to-severe PMS' and 'PMDD' according to the PSQ. The girls were classified into the 'absent' group if they were absent for more than 1 day per month. We used multivariate logistic analysis to examine the risk factors for school absenteeism. Results: The rates of 'moderate-to-severe PMS' and 'PMDD' were 9.9 and 3.1 %, respectively. A total of 107 girls (11.9 %) were classified into the 'absent' group. Significant differences were observed in the prevalence of all premenstrual symptoms (p < 0.001), 'age' (p < 0.001), 'a preference for salty food' (p = 0.001), and 'lack of regular exercise' (p = 0.03) between the 'absent' and 'non-absent' groups. Multivariate analysis revealed that premenstrual symptoms such as 'insomnia or hypersomnia' (odds ratio [OR] 2.27, 95 % confidence interval [CI]: 1.46-4.17) and 'physical symptoms' (OR 2.24, 95 % CI: 1.37-3.66), 'reduced social life activities' (OR 2.71, 95 % CI 1.31-5.59), and 'a preference for salty food' (OR 1.89, 95 % CI: 1.20-2.98) were risk factors for school absenteeism. Conclusions: One in nine Japanese female high school students were absent from school due to premenstrual symptoms. Physical premenstrual symptoms and lifestyles, such as a preference for salty food and a lack of regular exercise, were identified as risk factors for school absenteeism.

Objectives: To investigate the relationship between the occurrence of stress fracture and premenstrual syndrome (PMS)/premenstrual dysphoric disorder (PMDD) in Japanese adolescent athletes. Design: Cross-sectional study. Setting: Osaka, Japan. Participants: A school-based survey on menstruation and school life was conducted using a sample of 18-18 Japanese female students who belonged to two public high schools in Japan. Among them, we recruited 394 athletes who had regular menstrual cycles (25-38 days) and completed a questionnaire about their premenstrual symptoms and their competitive career. Main outcome measure: Premenstrual symptoms and the occurrence of stress fracture. Results: The prevalences of moderate-to-severe PMS and PMDD were 8.9% and 1.3%, respectively, which were the same as in collegiate athletes in a previous study. Premenstrual symptoms disturbed 'Work efficiency or productivity, home responsibilities', 'Relationships with coworkers or family' and 'Athletic performance in training or competition' more severely than menstrual pain (p = 0.031, p = 0.004 and p < 0.001, respectively). 66 athletes (16.8%) reported having experienced a stress fracture. The severity of 'Overeating or food cravings', 'Physical symptoms' and 'Performance in training or competition' in athletes with previous stress fractures were much higher than in those without a history of stress fractures (p = 0.015, p = 0.008 and p = 0.006, respectively). In terms of premenstrual symptoms, 'Physical symptoms' was associated with an increased risk of stress fractures in athletes (OR 1.66, 95% CI 1.06 to 2.62). Conclusions: The results from this study indicated that premenstrual symptoms may affect athletic performance and has the risk of stress fractures in adolescent athletes.

Objective: To examine psychological distress among pregnant women in Miyagi prefecture which was directly affected by the Great East Japan Earthquake and tsunami and compare other areas of Japan that were less damaged. Methods: This study was conducted in conjunction with the Japan Environment and Children's Study (JECS). We examined 10,129 Japanese women using the primary fixed data of the JECS. The Kessler 6-item psychological distress scale (K6) was administered to 7473 eligible women including 998 in Miyagi unit center ('Miyagi UC') and 6475 in the other unit centers ('13UC5'). We compared the prevalence and the risk of distress (K6 >13) during pregnancy in 'Miyagi UC' and '13UC5'. Results: More women in 'Miyagi UC' (4.9%) suffered psychological distress, compared with '13UC5' (3.1%) (p <0.001). A significantly higher prevalence of women in 'Miyagi UC' (55.5%) had experienced negative life events, whereas '13UCs' showed 42.7% (p < 0.0001). In multivariable logistic analyses adjusted for baseline characteristics, there was a significant regional difference of psychological distress (adjusted odds ratio; aOR in Miyagi UC= 1.488; 95%CI, 1.059-2.090). After further adjusting for negative life events, the association was diminished (aOR=1.338; 95%Cl, 0.949-1.884). Limitations: The JECS had no data before the earthquake and the extent of damage was not investigated. Possible regional representativeness is also a limitation. Conclusion: After the Great East Japan Earthquake, the prevalence of pregnant women with psychological distress (K6 >= 13) were high in Miyagi prefecture. Especially in the coastal area directly affected by tsunami, it is high with or without negative life events experienced. (C) 2015 Elsevier BY. All rights reserved.

AimA combined oral contraceptive containing ethinylestradiol 20 mu g plus drospirenone 3mg (EE20+DRSP) in a 24/4 regimen has been shown to alleviate the symptoms of premenstrual syndrome and premenstrual dysphoric disorder. This study was conducted to evaluate the efficacy of EE20+DRSP in Japanese patients with premenstrual symptoms. Material and MethodsA multicenter, prospective, open-label, single-arm, phase IV study was performed in Japanese women with dysmenorrhea and premenstrual symptoms. They were treated with EE20+DRSP to alleviate the symptoms of dysmenorrhea for six treatment cycles. Premenstrual symptoms were evaluated using a Premenstrual Symptoms Questionnaire at baseline and after three and six cycles of EE20+DRSP. The degree of dysmenorrhea was also evaluated using a visual analog scale at baseline and after one, three, and six cycles of EE20+DRSP. ResultsForty-eight patients were treated with EE20+DRSP. Most of the premenstrual symptoms were alleviated significantly by three and six cycles of EE20+DRSP treatment. EE20+DRSP treatment significantly improved the severity of premenstrual symptoms. We also confirmed the effectiveness of EE20+DRSP for the treatment for dysmenorrhea. ConclusionThis study showed that EE20+DRSP could be a useful treatment strategy for premenstrual symptoms in Japanese women.

Study Objective: To determine the prevalence and impact of premenstrual syndrome (PMS) and premenstrual dysphoric disorder (PMDD) in Japanese collegiate athletes, with a focus on their disruption of athletic performance. Design: Cross-sectional study. Setting: A university in Osaka, the largest city in western Japan. Participants: 232 female collegiate athletes. Main Outcome Measures: Premenstrual symptoms and social activities. Results: The prevalence of each premenstrual symptom was high. The prevalence of moderate to severe PMS and PMDD was 8.6% and 2.9%, respectively, the same as in general high school students. The athletic performance of 44.3% of athletes was found to suffer in a game or in practice. "Elite athletes" (OR 8.63,95% CI: 1.22-120.0), "Difficulty concentrating" (OR 3.15, 95% CI: 1.05-10.6), and "Fatigue or lack of energy" (OR 5.92, 95% CI: 1.32-34.5) increased the risk of poor athletic performance. Conclusions: This study showed that premenstrual symptoms affect not only the daily activities but also the athletic performance of collegiate athletes.

Background: Hachimijiogan (HJG), Ba-Wei-Di-Huang-Wan in Chinese, is one of the most popular herbal medicines in Japanese Kampo. HJG is often prescribed for the prevention and treatment of age-related diseases. Muscle atrophy plays an important role in aging-related disabilities such as sarcopenia. The purpose of this study was to investigate the possible beneficial effect of HJG on skeletal muscle. Methods: Cells of murine skeletal muscle myoblast cell line C2C12 were used as an in vitro model of muscle cell proliferation and differentiation. The effect of HJG on C2C12 cell proliferation and differentiation was assessed. We counted the number of myotubes morphologically to assess the degree of differentiation. Results: HJG treatment (200 mu g/mL) for 3 days significantly increased C2C12 cell number by 1.23-fold compared with that of the control. HJG promoted the proliferation of C2C12 cells through activation of the ERK1/2 signaling pathway without affecting the Akt signaling pathway. HJG did not affect the differentiation of C2C12 cells. Conclusion: HJG had beneficial effects on skeletal muscle myoblast proliferation. These findings may provide a useful intervention for the prevention and treatment of sarcopenia.

It is well known that glucocorticoid receptor (GR) signaling regulates the hypothalamic-pituitary-adrenal (HPA) axis, and GR expression level is associated with HPA axis activity. Recent studies revealed that microRNA-(miR-) 18 and/or 124a are candidate negative regulators of GR in the brain. The Kampo medicine Yokukansan (YKS) can affect psychological symptoms such as depression and anxiety that are associated with stress responses. In this study, we evaluated the effect of YKS on miR-18 and 124a and GR levels in mice exposed to stress. We found that YKS pretreatment normalized elevated plasma corticosterone levels in stress-exposed mice. In addition, GR mRNA levels were downregulated in the brain following stress exposure. While miR-124a expression levels were not altered in the hypothalamus of stress-exposed mice, miR-18 levels decreased in the hypothalamus of YKS-pretreated mice after stress exposure. Finally, GR protein levels in the paraventricular nucleus (PVN) of the hypothalamus after stress exposure recovered in YKS-pretreated mice. Collectively, these data suggest that YKS normalizes GR protein levels by regulating miR-18 expression in the hypothalamus, thus normalizing HPA axis activity following stress exposure.

Background: Hachimijiogan (HJG), Ba-Wei-Di-Huang-Wan in Chinese, is one of the most popular herbal medicines in Japanese Kampo. HJG is often prescribed for the prevention and treatment of age-related diseases. Muscle atrophy plays an important role in aging-related disabilities such as sarcopenia. The purpose of this study was to investigate the possible beneficial effect of HJG on skeletal muscle. Methods: Cells of murine skeletal muscle myoblast cell line C2C12 were used as an in vitro model of muscle cell proliferation and differentiation. The effect of HJG on C2C12 cell proliferation and differentiation was assessed. We counted the number of myotubes morphologically to assess the degree of differentiation. Results: HJG treatment (200 μg/mL) for 3 days significantly increased C2C12 cell number by 1.23-fold compared with that of the control. HJG promoted the proliferation of C2C12 cells through activation of the ERK1/2 signaling pathway without affecting the Akt signaling pathway. HJG did not affect the differentiation of C2C12 cells. Conclusion: HJG had beneficial effects on skeletal muscle myoblast proliferation. These findings may provide a useful intervention for the prevention and treatment of sarcopenia.

The aim of this study was to elucidate whether metformin can regulate the expression of vascular endothelial growth factor (VEGF) in rat-derived uterine leiomyoma cells (ELT-3 cells). In vitro studies were conducted using ELT-3 cells. Under normoxic conditions, metformin suppressed VEGF protein levels in the supernatant and cells in a dose-dependent manner. In hypoxia-mimicking conditions, VEGF and hypoxia-inducible factor-1 alpha (HIF-1 alpha) proteins were both highly expressed and were suppressed by the metformin treatment. Metformin did not affect HIF-1 alpha mRNA levels, which indicated that its effects occurred at the post-translational level. Metformin inhibited mammalian target of rapamycin complex 1 (mTORC1) activity by phosphorylating the mTORC1 component raptor. This study revealed the anti-angiogenic activity of metformin in ELT-3 cells by suppressing the expression of VEGF via the mTORC1/HIF-1 alpha pathway. These results indicate that metformin may represent an effective alternative in the future treatment of uterine leiomyomas. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

Uterine leiomyosarcomas generally do not respond well to standard chemotherapy. We previously demonstrated that curcumin, the active ingredient derived from the herb Curcuma longa, inhibits uterine leiomyosarcoma cells in vitro via the inhibition of the AKT-mammalian target of rapamycin (mTOR) pathway. As a preclinical investigation, we performed an in vivo study using female nude mice to confirm the therapeutic potential of curcumin against uterine leiomyosarcoma. Human leiomyosarcoma cells, SK-UT-1, were inoculated in female nude mice to establish subcutaneous tumors. Either vehicle control or 250 mg/kg curcumin was administered intraperitoneally every day for 14 consecutive days, and the mice were then killed. The tumors were measured every 2-3 days. The tumors were processed for immunohistochemical analyses to detect total AKT, phosphorylated AKT, total mTOR, phosphorylated mTOR, and phosphorylated S6. To detect apoptosis, the tumors were stained for cleaved PARP and TUNEL. Ki-67 immunohistochemistry was performed to determine cell viability of the tumors. Compared with the control, curcumin reduced uterine leiomyosarcoma tumor volume and mass significantly with a concordant decrease in mTOR and S6 phosphorylation. However, AKT phosphorylation was not significantly altered. Cleaved PARP and TUNEL staining increased significantly with curcumin administration, indicating the induction of apoptosis. There was no difference in Ki-67 staining between the two groups. Curcumin inhibited uterine leiomyosarcoma tumor growth in vivo by targeting the AKT-mTOR pathway for inhibition.

Study Objective: To determine the relationship between dysmenorrhea and posttraumatic stress disorder (PTSD) among Japanese adolescent girls 9 months after the Great East Japan Earthquake. Design: Cross-sectional study. Setting: Two high schools in Sendai, the largest city in northeastern Japan. Participants: 1489 female adolescents aged between 15 and 18 years. Main Outcome Measures: Impact of Event Scale-Revised (IES-R), intensity of menstrual pain. Results: The intensity of menstrual pain increased according to the comorbidity of PTSD (P < .001). The IES-R subscale and total scores significantly increased according to the severity of menstrual pain. Conclusions: This study showed a significant association between natural disaster-induced PTSD and the severity of dysmenorrhea.

On March 11, 2011, the Great East-Japan Earthquake occurred and a massive tsunami hit the northeastern coast of Japan. Catastrophic disasters such as earthquakes and war cause tremendous damage, not only physically but also mentally. Posttraumatic stress disorder (PTSD) is an anxiety disorder that occurs in the aftermath of a traumatic event. Premenstrual syndrome (PMS) is a cluster of psychological and somatic symptoms that are limited to the late luteal phase of the menstrual cycle. Premenstrual dysphoric disorder (PMDD) is considered a severe form of PMS. To determine the relationship between premenstrual symptoms and natural disaster-induced PTSD among Japanese adolescent girls, we conducted a cross-sectional study. Overall, 1489 high school students who belong to two high schools in Sendai, the largest city in northeastern Japan, were assessed 9 months after the earthquake. These schools are located inland, far from the seashore, and were not damaged by the tsunami. Premenstrual symptoms were assessed using the Premenstrual Symptoms Questionnaire, and PTSD symptoms were assessed using the Japanese-language version of Impact of Event Scale-Revised, which is a widely used self-assessment questionnaire about PTSD symptoms. We analyzed the data of 1,180 girls who completed the questionnaires and 118 girls (10.0%) were classified as having PTSD. The prevalence rates of PMDD and moderate to severe PMS increased according to the comorbidity of PTSD (p < 0.001), showing a correlation between the severity of PMS/PMDD and natural disaster-induced PTSD. The comorbidity of PMS/PMDD and PTSD may complicate the follow-up of both conditions.

Uterine leiomyosarcoma (LMS) has an unfavorable response to standard chemotherapeutic regimens. Two natural occurring compounds, curcumin and epigallocatechin gallate (EGCG), are reported to have anti-cancer activity. We previously reported that curcumin reduced uterine LMS cell proliferation by targeting the AKT-mTOR pathway. However, challenges remain in overcoming curcumin's low bioavailability. The human LMS cell line SKN was used. The effect of EGCG, curcumin or their combination on cell growth was detected by MTS assay. Their effect on AKT, mTOR, and S6 was detected by Western blotting. The induction of apoptosis was determined by Western blotting using cleaved-PARP specific antibody, caspase-3 activity and TUNEL assay. Intracellular curcumin level was determined by a spectrophotometric method. Antibody against EGCG cell surface receptor, 67-kDa laminin receptor (67LR), was used to investigate the role of the receptor in curcumin's increased potency by EGCG. In this study, we showed that the combination of EGCG and curcumin significantly reduced SKN cell proliferation more than either drug alone. The combination inhibited AKT, mTOR, and S6 phosphorylation, and induced apoptosis at a much lower curcumin concentration than previously reported. EGCG enhanced the incorporation of curcumin. 67LR antibody partially rescued cell proliferation suppression by the combination treatment, but was not involved in the EGCG-enhanced intracellular incorporation of curcumin. EGCG significantly lowered the concentration of curcumin required to inhibit the AKT-mTOR pathway, reduce cell proliferation and induce apoptosis in uterine LMS cells by enhancing intracellular incorporation of curcumin, but the process was independent of 67LR.

Objective: Uterine leiomyosarcoma (LMS) has an unfavorable response to standard chemotherapy. A natural occurring compound, curcumin, has been shown to have inhibitory effects on cancers. We previously demonstrated that curcumin reduced uterine LMS cell proliferation by targeting the AKT-mTOR pathway and activating apoptosis. To further explore the anticancer effect of curcumin, we investigated the efficacy of curcumin on autophagy in LMS cells. Methods: Cell proliferation in human uterine LMS cell lines, SKN and SK-UT-1, was assessed after exposure to rapamycin or curcumin. Autophagy was detected by Western blotting for light chain 3 and sequestosome 1 (SQSTM1/p62) expression. Apoptosis was confirmed by Western blotting for cleaved poly (ADP-ribose) polymerase (PARP). Results: Both rapamycin and curcumin potently inhibited SKN and SK-UT-1 cell proliferation in a dose-dependent manner. Curcumin induced autophagy and apoptosis in SKN and SK-UT-1 cells, whereas rapamycin, a specific mTOR inhibitor, did not. Curcumin increased extracellular signal-regulated kinase 1/2 activity in both SKN and SK-UT-1 cells, whereas PD98059, an MEK1 inhibitor, inhibited both the extracellular signal-regulated kinase 1/2 pathway and curcumin-induced autophagy. Conclusions: These experimental findings suggest that curcumin is a potent inhibitor of cell proliferation in uterine LMS and provide new insights about ongoing signaling events leading to the possible development of a new therapeutic agent.

Uterine leiomyomas are the most common gynecological benign tumors and greatly affect reproductive health and wellbeing. Metformin is the most widely used antidiabetic drug in the world, and there is increasing evidence of a potential efficacy of this agent as an anticancer drug. In order to understand metformin's anti-tumorigenic potential better, in this study, we investigated the inhibitory effect of metformin and expression of key targets of metformin cell signaling in leiomyoma cells. Cell proliferation was assessed after exposure to metformin. Apoptosis was assessed by western blotting for cleaved-PARP and TUNEL staining. The expressions of phosphorylated AMPK and phosphorylated S6 were determined by western blotting. Metformin potently inhibited ELT-3 cell proliferation in a dose-dependent manner. Western blotting analysis demonstrated that metformin induced phosphorylation of AMPK and the inhibitory effect was attenuated with AMPK inhibitor, compound C. In parallel, treatment with metformin decreased phosphorylation of S6 protein. These experimental findings show that metformin is a potent inhibitor of cell proliferation in leiomyoma cells. This effect is mediated by AMPK activation and subsequent inhibition of the mTOR pathway. Thus, this study provides a possible mechanism of the action of metformin in the inhibition of leiomyoma cell growth. © 2013 Informa UK, Ltd.

The objective of this study was to develop a questionnaire for the diagnosis of Qi stagnation. At first, we made the preliminary version of the questionnaire from 30 symptoms most frequently mentioned about Qi stagnation in classic books of Oriental Medicine. Two hundred and seven participants completed the preliminary version of the questionnaire rating the severity of 30 symptoms. Those participants were assessed for Qi stagnation by 2 physicians. Logistic regression analysis was performed between the physicians' assessment of Qi stagnation and the severity of symptoms in the preliminary questionnaire. The final version of the questionnaire was developed with 23 symptoms that had significant odds ratios. The Cronbach's alpha coefficient was 0.83. The area under the curve was 0.90 and cut-off value for diagnosis of Qi stagnation was 28.5 in receiver operating characteristic (ROC) analysis. Sensitivity and specificity were 0.83 and 0.80, respectively. The Spearman's correlation coefficient was 0.72 in the test-retest. This questionnaire would enable standardization and objective verification of the diagnosis of Qi stagnation. (c) 2011 Elsevier Ltd. All rights reserved.

Kampo (Japanese herbal) medicine is the complementary and alternative medicine that is most frequently used by Japanese doctors. We studied the perceptions and attitudes of Japanese gynecologic cancer patients to Kampo medicines and analyzed the characteristics of the backgrounds of Kampo users. A total of 476 patients with gynecologic cancer completed a self-reported questionnaire on Kampo medicine. State anxiety and trait anxiety were also assessed using the State-Trait Anxiety Inventory. It was confirmed that 22.9% of the women had used Kampo medicine. Kampo users were more likely to have had chemotherapy and were more likely to have experienced uncomfortable side effects of cancer treatment. Kampo users were more likely to believe that 'Kampo offers relief of symptoms,' 'fewer side effects than Western-style medicine,' and 'is not less effective than Western-style medicine' than nonusers. Kampo users expressed a stronger attitude of 'I want to take Kampo medicine.' Multiple risk ratio regression analysis revealed that chemotherapy (RR, 1.82; 95% CI, 1.14-2.91), lower state anxiety (RR, 0.76; 95% CI, 0.58-1.00), and higher trait anxiety (RR, 1.46; 95% CI, 1.11-1.92) were independently associated with Kampo use. This study showed that slightly less than one-fourth of Japanese gynecologic cancer patients take Kampo medicine. Kampo users made more favorable comments on Kampo medicine than nonusers. Our findings suggest that the psychological characteristics of individual patients is one of the factors that can influence the usage of Kampo.

To determine the relationship between premenstrual symptoms and dysmenorrhea among Japanese adolescent girls, a total of 1,431 high school students were assessed. Of them, 11.3% were classified with "moderate to severe premenstrual syndrome (PMS)" and 3.2% with " premenstrual dysphoric disorder (PMDD). "Eighty-five percent of the girls had dysmenorrhea. The rates of prevalence of PMDD and moderate to severe PMS were increased according to the severity of dysmenorrhea (rs=0.479), showing a correlation between the severity of PMS/PMDD and dysmenorrhea in adolescents.

Gynecology in the office setting is developing worldwide. Clinical guidelines for office gynecology were first published by the Japan Society of Obstetrics and Gynecology and the Japan Association of Obstetricians and Gynecologists in 2011. These guidelines include a total of 72 clinical questions covering four areas (Infectious disease, Malignancies and benign tumors, Endocrinology and infertility, and Healthcare for women). These clinical questions were followed by several answers, backgrounds, explanations and references covering common problems and questions encountered in office gynecology. Each answer with a recommendation level of A, B or C has been prepared based principally on evidence or consensus among Japanese gynecologists.These guidelines would promote a better understanding of the current standard care practices for gynecologic outpatients in Japan.

Purpose: Traditionally, oral estrogens have been used for hormone replacement therapy. However, in Japan, additional estrogen formulations have been used, including transdermal patches and transdermal gels. The latter have a unique commonality with cosmetics because both of them are applied to the skin. Beauty care is one of the most important lifestyle factors for women, and it has been reported that the amount of attention paid to beauty care has an effect in determining whether or not women will choose to undergo HRT during menopause. Therefore, our study focused on estrogen formulations and beauty care practices. Patients and methods: Fifty women who use hormone replacement therapy were recruited from the outpatient clinic of Tohoku University Hospital. They were treated with oral conjugated estrogen (n = 11), transdermal 17β-estradiol patch (n = 11), and transdermal 17β-estradiol gel (n = 28). They completed a questionnaire to assess their lifestyle (beauty care practices and exercise habits) and their compliance. The transdermal gel users were further interviewed about their subjective impressions regarding "smell", "sticky feeling", "spreadability", and "irritation" on the skin using a five-grade scale. Results: There were no differences in the usability of medicines and patient compliance among the estrogen formulations. We observed a positive tendency between the level of beauty care and transdermal gel use (P = 0.0645, ordinary logistic regression analysis). The gel users placed top priority on a lack of "sticky feeling" but the subjective impression regarding "sticky feeling" was worst among the four factors (P, 0.01, Steel-Dwass test). Correspondence analysis showed that the subjective impressions of transdermal gel corresponding to usability in the range of "moderate" to "very good" and "sticky feeling" greatly affected the usability of the formulation. Conclusion: These results suggest that the level of attention to beauty care plays some role in the choice of estrogen formulations. © 2012 Takeda et al.

The Great East Japan earthquake and tsunami disaster that occurred on March 11, 2011 seriously destroyed Japanese social activities the medical system included. We provided medical support to the damaged area, and mainly performed Oriental medicine. Traditional methods using physical diagnoses and the treatments with herbs, acupuncture, and massage were effective, where any infrastructure had suffered or any modern medical facilities had been destroyed. Acute phase infectious disease, common colds, and hypothermia were dominant. Allergies increased two weeks later, and there was much mental distress, and chronic pain symptoms one month later. We prescribed Kampo herbal medicines for common colds, hypothermia, allergies, and mental distress. Moreover, we also performed acupuncture and kneaded patients' body to reduce pain, stiffness, and edema. These treatments were effective for both physical and mental distress. Thus we believe that Oriental medicine is valuable in disaster situations.

Objective. Uterine leiomyomas are the most common gynaecological benign tumour and greatly affect reproductive health and wellbeing. They are the predominant indication for hysterectomy in premenopausal women. Curcumin, a well-known component of turmeric, has been reported to prevent various diseases such as cancer, diabetes and obesity. Previous study reported that curcumin represses the proliferation of several tumour cells. However, there has not been a precise characterisation of the curcumin-induced inhibition of uterine leiomyoma cells. In this study, we investigated the inhibitory effect of curcumin on leiomyoma cells proliferation. Study design. aEuro integral Eker rat-derived uterine leiomyoma cell lines (ELT-3 cells) were used. Cell proliferation was assessed by counting the number of cells and MTS assay. The activation of peroxisome proliferator-activated receptor-gamma (PPAR gamma gamma) was evaluated by luciferase assay. Results. aEuro integral We found that curcumin significantly inhibited ELT-3 cell proliferation. PPAR gamma gamma was expressed in ELT-3 cells and curcumin acted as a PPAR gamma gamma ligand. This inhibitory effect of curcumin was attenuated by the treatment of cells with PPAR gamma gamma antagonist. Conclusion. aEuro integral These experimental findings in vitro show that the inhibitory effect of curcumin on ELT-3 cell proliferation occurs through the activation of PPAR gamma gamma. Curcumin may be useful as an alternative therapy for uterine leiomyoma.

Objective. Uterine leiomyosarcoma generally has an unfavorable response to standard chemotherapy. The loss of PI EN which results in constitutive AKT-mTOR activation causes an increase in leiomyosarcoma formation in mice. The active ingredient derived from the herb Curcuma longa, curcumin, shows antitumor properties in a variety of cancer cell lines by altering a number of oncogenic pathways. To explore the possibility of curcumin as an alternative to standard chemotherapy, we decided to investigate curcumin's antitumor effect on uterine leiomyosarcoma cells. Methods. Human leiomyosarcoma cell lines, SKN and SK-UT-1, were cultured for in vitro experiments. Rapamycin or curcumin was added in different doses and their effect on cell growth was detected by MTS assay. The influence of rapamycin or curcumin on ART, mTOR, p70S6 and S6 phosphorylation and protein expression was detected by Western Blotting. The ability of rapamycin or curcumin to induce apoptosis was determined by Western blotting using cleaved-PARP specific antibody, Caspase-3 activity assay and TUNEL assay. Results. Both rapamycin and curcumin significantly reduced SKN cell proliferation. Curcumin inhibited mTOR, p70S6 and S6 phosphorylation similar with rapamycin. Cleaved PARP, caspase-3 activity and DNA fragmentation increased proportional with curcumin concentration. At a high concentration, curcumin significantly induced apoptosis in SKN cells, but not rapamycin. Conclusions. Curcumin inhibited uterine leiomyosarcoma cells' growth by targeting the AKT-mTOR pathway for inhibition. However, rapamycin, a specific mTOR inhibitor, did not induce apoptosis in SKN cells unlike curcumin that also has a pro-apoptotic potential in SKN cells. (C) 2011 Elsevier Inc. All rights reserved.

In traditional Chinese medicine, moxibustion is a local thermal therapy that is used for several conditions. Quantifying the effects of moxibustion therapy has been difficult because the treatment temperature depends on the physician's experience, and the temperature distribution in the target area is not uniform. This prospective observational study aims to quantify the effect of local thermal stimulation to the abdomen. We developed a heat transfer control device (HTCD) for local thermal stimulation. Twenty-four healthy subjects were enrolled and they underwent abdominal thermal stimulation to the para-umbilical region with the device for 20 min. Blood flow volume in the superior mesenteric artery (SMA) and brachial artery (BA), the heart rate and the blood pressure were measured at rest, 15 min after starting thermal stimulation and 10, 20, 30 and 40 min after completing thermal stimulation. Blood flow parameters were measured by high-resolution ultrasound. In the SMA, blood flow volume was significantly increased during thermal stimulation (P < .01), as well as at 10 min (P < .01) and 20 min (P < .05) after stimulation. In the BA, blood flow volume decreased at 40 min after stimulation (P < .01). In conclusion we could quantify the effect of local thermal stimulation with an HTCD and high-resolution ultrasound. Thermal stimulation of the para-umbilical region increased blood flow in the SMA 20 min after stimulation in healthy subjects.

Objective. Although fibroids greatly affect reproductive health, the pathophysiology and epidemiology are not well known. Recently, we have reported the relationship between uterine leiomyoma and metabolic syndrome. Many studies have indicated that reductions in plasma adiponectin levels play major roles in the development of metabolic syndrome. In this study, we investigated the significant repressive effect of adiponectin on rat uterine leiomyoma ELT-3 cells proliferation. Study design. Expression of adiponectin receptor 1 and receptor 2 was evaluated by RT-PCR and Western blot analysis. Cell proliferation was assessed by the MTS assay and cell counting. Apoptosis was evaluated by Hoechst staining and cleaved poly (ADP-ribose) polymerase (PARP). Results. Adiponectin receptor 1 and receptor 2 were expressed in ELT-3 cells. Adiponectin repressed rat uterine leiomyoma ELT-3 cells cell proliferation without inducing apoptosis. Conclusion. The repression of adiponectin on leiomyoma cell proliferation in the rat may explain a crucial role of adiponectin in the association of metabolic syndrome with uterine leiomyoma.

To determine the prevalence and the impact of premenstrual symptoms among Japanese adolescent girls, a total of 618 high school students were assessed. Of them, 64.6% were found to suffer from premenstrual symptoms, which is lower than that in adult women. On the other hand, the rates of prevalence of moderate to severe PMS and PMDD in girls were higher than those in adult women. Premenstrual symptoms could have significant consequences by interfering with the daily functioning of adolescent girls.

Uterine leiomyomas are the most common gynecological benign tumor and greatly affect reproductive health and wellbeing, but the pathophysiology and epidemiology of uterine leiomyoma are poorly understood. One of the major reasons for the slow progress in leiomyoma research is the lack of a good in vivo model system. We therefore aimed to develop a novel model by transplanting human uterine leiomyoma xenografts in an immunodeficient mouse strain (NOD/SCID/gamma c-null: NOG). Human uterine leiomyoma tissues were cut into small pieces and inserted subcutaneously into the right and left flanks of NOG mice. Estrogen supplementation was needed to maintain the features of uterine leiomyoma in xenografted tissues. After 4 weeks or 8 weeks of transplantation, xenografted tissues were harvested and analyzed regarding tissue morphology, collagen content, and proliferation and apoptosis of uterine leiomyoma smooth muscle cells. The xenografts that were harvested after 4 weeks and 8 weeks retained the histological architecture of original uterine leiomyoma tissue both in cellular and collagen components. The expression profiles of key markers of uterine leiomyoma were also maintained, including estrogen receptor, progesterone receptor, and a-smooth muscle actin, as judged by immunohistochemical staining. The proportion of proliferating cells was significantly increased (1.5-fold) in the xenografts after 8 weeks of transplantation, whereas that of the apoptotic cells remained unchanged. Importantly, the reproducible results were obtained with the tumor tissues derived from six patients. The present in vivo model may provide a useful tool for development of novel therapeutic strategies for uterine leiomyoma.

Introduction: The aim of this study was to identify the prognostic factors and to establish a model for the prediction of life expectancy in patients with recurrent cervical cancer that had previously been treated with radiotherapy. Methods: The records of consecutive women with recurrent cervical cancer after radiotherapy were retrospectively reviewed. Primary disease, follow-up, and recurrence data were collected. Univariate and multivariate analyses of prognostic factors of survival were performed. Results: A total of 162 patients were included in our database. The median survival after recurrence was 15 months. Multivariate analysis revealed that symptom status, the site of relapse, prior chemoradiotherapy, and treatment modality were significant prognostic factors in terms of survival after recurrence. Patient survival was inversely correlated with the number of these prognostic factors. When the patients were divided into 3 prognostic groups, (low risk: patients with no poor prognostic factors; intermediate: patients with one poor prognostic factor; and high-risk: patients with more than 2 poor prognostic factors), the patients in the high-risk group had a significantly shorter survival (median, 10 months) than those with one risk factor (median, 20 months) or no risk factors (median, 36 months). Conclusions: Symptom status, the site of relapse, prior chemoradiotherapy, and treatment modality are significant prognostic factors in patients with recurrent cervical cancer that had previously been treated with radiotherapy. Our prognostic model, composed of 4 clinical variables, may enable physicians to predict survival more accurately.

Study design. This study investigated the potential role of aldosterone in the proliferation of ELT-3 leiomyoma cells. Results. Aldosterone-induced ELT-3 leiomyoma cell proliferation and the expression of mineralocorticoid receptor (MR) were confirmed. Pre-incubating the cells with the MR blockers spironolactone or eplerenone effectively repressed aldosterone-induced and angiotensin II (Ang II)-induced cell proliferation. Treatment of aldosterone increased the levels of Ang II type-1 receptor mRNA. Conclusion. These experimental findings in vitro show the presence of complex regulation of Ang II and aldosterone induced leiomyoma cell proliferation.

Objectives: The aim of this study was to evaluate whether nedaplatin-based concurrent chemoradiotherapy (CCRT) using high-dose-rate intracavitary brachytherapy (HDR-ICBT) is superior to radiotherapy (RT) alone in patients with FIGO stage IIIb cervical cancer. Methods: The records of 41 consecutive women treated either with nedaplatin-based CCRT using HDR-ICBT (n = 20) or RT alone (nonrandomized control group, n = 21) for stage IIIb cervical cancer were retrospectively reviewed. The activity and toxicity were compared between the two treatment groups. Progression-free survival (PFS) and overall survival (OS) were the main endpoints. Results: The 5-year overall survival rates in the CCRT and RT groups were 65 and 33.3%, respectively. The median OS of the CCRT and RT groups were 60 and 29 months, respectively. CCRT was significantly superior to RT alone with regard to PFS (p = 0.0015) and OS (p = 0.0364). The frequency of acute grade 3-4 toxicity was significantly higher in the CCRT group than in the RT group. However, no statistically significant difference was observed with regard to severe late toxicity. Conclusions: Nedaplatin-based concurrent chemoradiotherapy was safely performed and significantly improved the prognosis of patients with FIGO stage IIIb cervical cancer. This treatment can be considered as an alternative to cisplatin-based chemoradiotherapy in this patient population. Copyright (C) 2010 S. Karger AG, Basel

Background: Diet is considered an important factor for bone health, but is composed of a wide variety of foods containing complex combinations of nutrients. Therefore we investigated the relationship between dietary patterns and fall-related fractures in the elderly. Methods: We designed a population-based prospective survey of 1178 elderly people in Japan in 2002. Dietary intake was assessed with a 75-item food frequency questionnaire (FFQ), from which dietary patterns were created by factor analysis from 27 food groups. The frequency of fall-related fracture was investigated based on insurance claim records from 2002 until 2006. The relationship between the incidence of fall-related fracture and modifiable factors, including dietary patterns, were examined. The Cox proportional hazards regression model was used to examine the relationships between dietary patterns and incidence of fall-related fracture with adjustment for age, gender, Body Mass Index (BMI) and energy intake. Results: Among 877 participants who agreed to a 4 year follow-up, 28 suffered from a fall-related fracture. Three dietary patterns were identified: mainly vegetable, mainly meat and mainly traditional Japanese. The moderately confirmed (see statistical methods) groups with a Meat pattern showed a reduced risk of fall-related fracture (Hazard ratio = 0.36, 95% CI = 0.13 - 0.94) after adjustment for age, gender, BMI and energy intake. The Vegetable pattern showed a significant risk increase (Hazard ratio = 2.67, 95% CI = 1.03 - 6.90) after adjustment for age, gender and BMI. The Traditional Japanese pattern had no relationship to the risk of fall-related fracture. Conclusions: The results of this study have the potential to reduce fall-related fracture risk in elderly Japanese. The results should be interpreted in light of the overall low meat intake of the Japanese population.

Objectives. The aim Of this Study was to evaluate the efficacy of postoperative nedaplatin-based concurrent chemoradiotherapy (CCRT) in patients with FIGO stage IA2-IIB cervical cancer with adverse risk factors. Methods. We retrospectively reviewed the medical records of 183 patients with early-stage cervical cancer who had undergone radical Surgery between April 1997 and March 2006. Of these, 68 patients displayed high-risk prognostic factors Such as positive pelvic lymph nodes, parametrial involvement, or a positive surgical margin. Fifty-seven patients demonstrated intermediate-risk prognostic factors including deep stromal invasion, capillary lymphatic space involvement, or large tumor diameter. These patients were treated postoperatively with CCRT or radiotherapy alone (RT). Fifty-eight patients showed no risk factors and, therefore, received no adjuvant therapy after Surgery. The 3-year recurrence rate, progression free survival (PFS), and overall Survival (OS) were compared between the treatment groups. Results. CCRT was significantly superior to RT alone with regard to recurrence rate, PFS, and OS in patients that displayed high-risk and intermediate-risk prognostic factors. The frequencies Of acute grade 3-4 toxicities were significantly higher in patients treated with CCRT than in those treated with RT alone. However, no statistically significant difference was observed with regard to severe late toxicities. Conclusions. Postoperative nedaplatin-based CCRT was safely performed and improved the prognosis of FIGO stage IA2-IIB cervical cancer patients displaying high-risk or intermediate-risk prognostic factors. This treatment can be considered as an alternative to cisplatin-based chemoradiotherapy in this patient population. (C) 2009 Elsevier Inc. All rights reserved.

Daikenchuto is a traditional herbal medicine that is used for the treatment of cold feeling in the abdomen, while Orengedokuto, also a traditional herbal medicine, is used for treating inflammatory and ulcerative diseases affecting internal organs. However, the effects of these herbal medicines on cardiac output (CO) and intestinal blood flow have never been investigated. This examiner-blinded randomized crossover study intended to clarify the influence of Daikenchuto and Orengedokuto on CO and blood flow volume in the superior mesenteric artery (SMA). Fourteen healthy men (35 +/- 7 years old) were randomly assigned to two groups: group A and group B. Initially, all subjects were given 50 ml of water orally. After 7 days, subjects in group A were given 5.0 g of Daikenchuto, and 7 days later they were given 2.5 g of Orengedokuto. These herbal medicines were given to group B subjects in the reverse order. CO and SMA blood flow volume were measured from rest to 90 min after the administration of water or each medicine. There was a significant increase in SMA blood flow volume after the administration of Daikenchuto, compared to water alone (p < 0.05) and Orengedokuto (p < 0.05). SMA blood flow volume was significantly increased between 5 and 90 min after administration of Daikenchuto (p < 0.01) compared to the resting state. However, there was no significant change in CO after the administration of either agent. The present study indicates that Daikenchuto increases SMA blood flow volume without increasing CO.

During early pregnancy, cytotrophoblast cells differentiate into extravillous trophoblast (EVT) cells and invade the uterine spiral arteries. This physiological process is essential for the development of maternal-fetal circulation. Because EVT cells are exposed to a low-oxygen environment during this process, we investigated the role of hypoxia in the mechanism that regulates the invasive behavior of EVT cells. Real-time PCR and immunofluorescent analysis were performed to investigate how hypoxia influences the expression of E-cadherin in villous explants cultures and in trophoblast-derived BeWo cells. We determined that hypoxia induced E-cadherin down-regulation through Snail up-regulation in villous explant cultures. The influence of E-cadherin loss was examined by analyzing the expression of alpha(5)-integrin and phosphorylated focal adhesion kinase (FAK) by Western blot and evaluating trophoblast invasion using a matrigel invasion assay. E-cadherin loss induced the up-regulation of alpha(5)-integrin, which leads to the tyrosine phosphorylation of FAK, resulting in an increase in the invasive activity of EVT cells. An alpha(5)-integrin neutralizing antibody inhibited the invasion of EVT cells by attenuating FAK tyrosine phosphorylation. Immunohistochemical analysis using clinical placental bed biopsies revealed that alpha(5)-integrin was up-regulated and FAK tyrosine phosphorylated (Try(861)) as EVT cells invade the uterine myometrium, whereas E-cadherin expression was down-regulated. These results suggest that alpha(5)-integrin up-regulation induced by E-cadherin loss under hypoxia has a crucial role in regulating the migration of EVT cells. This finding should help us reach a better understanding of the pathogenesis of critical gestational diseases, such as preeclampsia. (Endocrinology 150: 4306-4315, 2009)

Placenta growth factor (PlGF) is a placental angiogenic factor. Metal-responsive transcription factor (MTF)-1 was reported to take part in the hypoxic induction of PlGF in RAS-transformed mouse fibroblasts. We contrarily showed that PlGF mRNA and protein levels decreased under hypoxia in a choriocarcinoma BeWo cell line derived from trophoblast. In this report, we examined whether hypoxia-dependent regulation of the PlGF gene in these cells also depends on MTF-1. We analyzed the effect of hypoxia on MTF-1 expression, and it was revealed to be decreased. Moreover, MTF-1 small interfering RNA treatment decreased PlGF mRNA level. To investigate the transcription of PlGF under hypoxia, we cloned promoter region of the human PlGF. Promoter deletion analysis suggested that triple repeats of metal-responsive element located between -511 and -468 bp in the promoter are important for the hypoxic regulation of PlGF. Treatment with MTF-1 small interfering RNA resulted in the significant decreased luciferase activity in PlGF reporter constructs. Chromatin immunoprecipitation showed the binding of the MTF-1 protein to the promoter region. We examined MTF-1 immunoreactivity in trophoblasts of term placental tissue from patients with normal pregnancies and preeclampsia, which represents a condition of placental hypoxia. Immunoreactivity of the MTF-1 protein was decreased in placentas from pregnant women with preeclampsia when compared with those from normal pregnant women. Taken together, these findings suggest that MTF-1 is involved in hypoxia-dependent regulation of PlGF in trophoblast-derived cells. (Endocrinology 150: 1801-1808, 2009)

Objective. Uterine leiomyomas are the most common gynecological benign tumor and greatly affect reproductive health and well-being. They are the predominant indication for hysterectomy in premenopausal women. Current epidemiological study reported that soy products intake is inversely associated with diseases leading to hysterectomy. Genistein is a soy-derived phytoestrogen and its inhibitory effect on leiomyoma cell proliferation is reported. In this study, we investigated the siginificant inhibitory effect of genistein on estradiol (E(2))-induced leiomyoma cells proliferation. Study design. The Eker rat-derived uterine leiomyoma cell line ELT-3 cells were used. Cell proliferation was assessed by counting the number of cells. The expression of estrogen receptors and peroxisome proliferator-activated receptor-gamma (PPAR gamma) was evaluated by Western blot analysis. Results. PPAR gamma was expressed in ELT-3 cells and genistein acted as PPAR gamma ligand. This inhibitory effect of genistein was attenuated by the treatment of cells with PPAR gamma antagonist bisphenol A diglycidyl ether ( BADGE) or GW9662. Conclusion. These experimental findings in vitro show that the repressive effect of genistein on E(2)-induced ELT-3 cell proliferation is through the activation of PPAR gamma. Genistein may be useful as an alternative therapy for leiomyoma.

BACKGROUND: Leiomyomatosis peritonealis disseminata is a rare disease characterized by multifocal proliferation of smooth muscle-like cells that are histologically similar to uterine leiomyoma. High levels of female gonadal steroids are considered to play an important role in the pathogenesis of leiomyomatosis peritonealis disseminata. CASE: A 68-year-old woman was referred to our hospital for pelvic tumor and multiple intraabdominal solid masses. After laparotomy, she was diagnosed as having leiomyomatosis peritonealis disseminata. Most of the disseminated tumors were nonresectable. Raloxifene was ineffective for the control of these tumors. Anastrozole was effective for the control of tumor growth and her symptoms. CONCLUSION: Anastrozole was useful for the management of postmenopausal nonresectable leiomyomatosis peritonealis disseminata. It is possible that the condition produces estrogen in situ, which promotes its growth in an autocrine/paracrine fashion.

Background: Radiation-induced enteritis is a serious clinical problem for which there is currently no recommended standard management. Daikenchuto (DKT) is a Japanese herbal medicine that has been used to treat adhesive bowel obstruction in Japan. This report describes a patient with radiation-induced enteritis whose clinical symptoms were much improved by treatment with DKT. Method: The patient was administered DKT, a traditional Japanese herbal formula, orally (2.5 g 3 times daily). Abdominal distention was evaluated objectively with computed tomography. Results: Gastrointestinal symptoms associated with radiation-induced enteritis were controlled successfully with DKT treatment. Conclusions: DKT treatment may be useful for the management of radiation-induced enteritis.

BACKGROUND: Although uterine leiomyomas or fibroids are the most common gynecological benign tumor and greatly affect reproductive health and well-being, the pathophysiology and epidemiology of uterine leiomyomas are poorly understood. Elevated blood pressure has an independent, positive association with risk for clinically detected uterine leiomyoma. Angiotensin II (Ang II) is a key biological peptide in the renin-angiotensin system that regulates blood pressure. METHODS: In this study, we investigated the potential role of Ang II (1-1000 nM) in the proliferation of rat ELT-3 leiomyoma cells in vitro. RT-PCR and western blot analysis with cell proliferation and DNA transfection assays were performed to determine the mechanism of action of Ang II. RESULTS: Ang II induced ELT-3 leiomyoma cell proliferation (P < 0.01) and the expression of Ang II type 1 receptor (AT(1)R) and AT(2)R mRNA and protein was confirmed. Regarding the intracellular signaling pathway, the Ang II-induced cell proliferation was AT(1)R-, epidermal growth factor receptor-, extracellular-regulated kinase-and protein kinase C-dependent but was not dependent on the AT2R or phosphatidylinositol-3 kinase or JAK kinase. The AT1R blocker telmisartan, effectively repressed Ang II-induced and estradiol-induced cell proliferation (P < 0.01). AT1R, but not AT2R, plays a role in Ang II-induced ELT-3 cell proliferation. CONCLUSIONS: These experimental findings in vitro highlight the potential role of Ang II in the proliferation of leiomyoma cells.

Background/Aims: Uterine leiomyomas are the most common gynecological benign tumor and greatly affect reproductive health and well-being. The pathophysiology and epidemiology of fibroids are poorly understood. Obesity and elevated blood pressure have been reported to be predisposing factors. In this study, we investigated whether fibroids are associated with some criteria of the metabolic syndrome. Methods: The case patients were 213 women who underwent hysterectomy or myomectomy for fibroids, and the control subjects were 159 women who underwent operation for benign indications other than fibroids. Preoperative information on body mass index (BMI), blood pressure (BP), serum triglyceride (TG) and fasting plasma glucose (FPG) was obtained from medical records. The patients were classified as overweight if they had a preoperatively measured BMI of >= 24.0, hypertensive if BP was >= 140/90 mm Hg, hypertriglyceridemic if TG was >= 150 mg/dl, and hyperglycemic if FPG was >= 110 mg/dl. Results: BMI, BP, TG and FPG were significantly higher in the case group compared with the control group. In logistic regression analysis, fibroids were statistically significantly associated with being overweight and hypertensive. With the combination of these risk factors, the risk of fibroids increased. Conclusion: Uterine leiomyomas may share pathogenic features with the development of metabolic syndrome. Copyright (C) 2008 S. Karger AG, Basel.

Objective. Uterine leiomyomata are the most common gynecological benign tumor and greatly affect reproductive health and well-being. The pathophysiology and epidemiology of fibroids are poorly understood. Gonadotropin-releasing hormone agonist (GnRH-a) pretreatment is one of the unfavourable factors for leiomyomata treatment with uterine artery embolisation (UAE). In this study, we investigated the plasma level of vascular endothelial growth factor (VEGF) in uterine leiomyoma patients with or without GnRH-a pretreatment. Study design. Thirty-two women who underwent UAE for symptomatic uterine leiomyoma were analysed. The plasma level of VEGF was studied before UAE. Results. The level of plasma VEGF was significantly higher in the GnRH-a pretreated group compared with the non-treated group. Conclusion. A compensative reaction for vasculature after GnRH-a treatment is speculated. Higher level of VEGF in GnRH-a pretreatment group could be one of the unfavourable factors for the treatment of uterine leiomyomata by UAE.

Glucose transporter-1 (GLUT1), one of the key functional indicators of placental differentiation, has an important role in placental glucose transport. We previously showed that the protein levels of GLUT1and nuclear transcription factor specificity protein-1 (Sp1) in rat choriocarcinoma cells (Rcho-1 cells) decreased during the differentiation of these cells to giant cells. We also showed that Sp1 was involved in the regulation of GLUT1 gene expression during this process. RelAassociated inhibitor (RAI) is an inhibitor of nuclear factor-kappa B that was identified by a yeast two-hybrid screen and is preferably expressed in human placenta and heart. RAI was also found to interact with Sp1 and exert an inhibitory effect against the DNA-binding activity of Sp1. We first show here that RAI mRNA expression increased as gestation proceeded and that RAI was localized mainly in the syncytiotrophoblast throughout pregnancy. The chloramphenicol acetyltransferase activity assay in Rcho-1 cells revealed that cotransfection of RAI expression vector resulted in decreased activity of the rat GLUT1 promoter but not in that of a mutated rat GLUT1 promoter lacking the Sp1 binding site. Furthermore, the protein level of RAI increased during differentiation. In addition, transfection of RAI expression vector promoted the morphological differentiation of Rcho-1 cells, and RAI knockdown using RAI-specific small interfering RNA reveals inhibitory effects on the morphological differentiation, as assessed by photomicroscopy. Taken together, these findings suggest that RAI may be involved in the regulation of trophoblast differentiation via interaction with Sp1.

Id-1, a member of the helix-loop-helix transcription factor family, inhibits the differentiation of Rcho-1 cells, which were derived from rat choriocarcinoma and consist of trophoblast stem cells. Id-1 is expressed at a high level in undifferentiated trophoblast stem cells and then downregulated during early differentiation, and is thought to be a key regulator in the trophoblast giant-cell differentiation pathway. In this study, we analyzed the signaling mechanism regulating the high expression levels of Id-1 in undifferentiated Rcho-1 cells. Promoter deletion analysis revealed that a 31-bp sequence (Box-2 region), located between -200 and -169 bp in the Id-1 promoter is necessary for the promoter activity. Electrophoretic mobility shift assays and DNA affinity precipitation assays showed that Box-2-binding activity was decreased during differentiation and that Sp-1 protein bound to this sequence. The protein level of Sp-1 was decreased during the differentiation. These results suggest that the Sp-1 protein level may regulate the Box-2-binding activity and the trophoblast giant-cell differentiation. (c) 2006 Elsevier Ltd. All rights reserved.

Background: A hypoxic environment is known to be essential for early placentation. A low oxygen tension induces hypoxia- inducible factor-1 ( HIF-1 alpha) which may play an important role as a transcription factor in maintaining the proliferative and undifferentiated phenotype in human trophoblasts. Methods: We analyzed the effect of a low oxygen tension on the rat trophoblast giant cell differentiation pathway using Rcho-1 cells which were derived from rat choriocarcinomas and consist of trophoblast stem cells. Results: We found that a low oxygen tension suppressed the morphological changes and steroidogenesis during differentiation. The anticipated downregulation of the ld-1 transcription factor, a negative regulator of trophoblast giant cell differentiation, was not observed in the hypoxic environment. On the other hand, deferoxamine, which mimics hypoxia and induces HIF-1 alpha, caused downregulation of ld-1 transcription factor and trophoblast giant cell differentiation. Conclusion: These results indicate that hypoxia represses rat trophoblast giant cell differentiation via an HIF-1 alpha -independent pathway. Copyright (c) 2007 S. Karger AG, Basel

In the trophoblast, constitutive expression of SOCS3 is important for the negative regulation of trophoblast giant cell differentiation. In this study, we analyzed the signaling pathway regulating the constitutive SOCS3 expression in undifferentiated Rcho-1 cells, which were derived from rat choriocarcinoma and consist of trophoblast stem cells that are capable of differentiating to trophoblast giant cells in vitro. PD98059, an MEK inhibitor, repressed the SOCS3 expression but AG490, a JAK2 inhibitor, did not. Promoter deletion analysis revealed that the STAT response element (SRE) in the SOCS3 promoter is necessary for the promoter activity. Overexpression of STAT3 increased the SOCS3 promoter activity, whereas expression of dominant-negative STAT3 reduced it. Constitutive STAT3 tyrosine phosphorylation that was not inhibited by either AG490 or PD98059 was demonstrated. Electrophoretic mobility shift assays showed the existence of a protein that bound to SRE and was supershifted with STAT3 antibody. This binding reaction was inhibited by neither AG490 nor PD98059. These findings imply that the ERK/MAPK pathway and STAT3 are involved in the constitutive activation of SOCS3 in undifferentiated Rcho-1 cells. Moreover, they indicate that the constitutive STAT3 tyrosine phosphorylation and the DNA binding activity of STAT3 do not depend on the ERK/MAPK or JAK kinase pathway. These results suggest that a trophoblast-specific STAT3 activation pathway is important for the regulation of giant cell differentiation.

To investigate the prevalence and impact of premenstrual symptoms in Japanese women, we developed the PSQ "The Premenstrual Symptoms Questionnaire" for the screening of premenstrual symptoms. The PSQ translates DSM-IV criteria into a rating scale with degrees of severity. One thousand one hundred and eighty-seven Japanese women between the ages of 20 and 49 yrs, who were seen at a clinic for uterine cancer screening, were assessed regarding their premenstrual symptoms using the PSQ. As many as 95% of these women were found to suffer from premenstrual symptoms. The rates of prevalence of moderate to severe PMS and PMDD in Japanese women were 5.3 and 1.2%, respectively, which are lower than those in Western women. Only 5.3% of women with moderate to severe PMS and PMDD were treated. The results of this study suggest that race and ethnicity influence the expression of premenstrual symptoms and that the current state of medical care for Japanese women with moderate to severe PMS and PMDD is not satisfactory.

Purpose: To compare the results of high dose rate (HDR) (Ir-192) and medium dose rate (MDR) (Cs-137) intracavitary brachytherapy (ICRT) for carcinoma of the uterine cervix. Materials and Methods: Between May 1991 and March 2001, a total of 206 patients with Stage I-IVA previously untreated cervical cancer were treated with ICRT combined with external beam radiotherapy (EBRT). HDR was administered to a total of 135 patients: 22 patients in Stage I, 49 in Stage II, 56 in Stage III, and eight in Stage IVA. MDR was administered to a total of 71 patients: six patients in Stage I, 27 in Stage II, 33 in Stage III, and five in Stage IVA. The MDR at point A was 30 Gy/hour for HDR and 1.7 Gy/hour for MDR treatment, and the corresponding median follow-up periods for survivors were 55 and 68 months. Results: For the HDR group, 5-year cause-specific survival rates were 90%, 78%, 53% and 33% for Stages I, II, III, and IVA, respectively. For the MDR group, the corresponding rates were 100%, 76%, 51%, and 40%. In the HDR group, 19 patients (14%) developed Grade 2 or higher late complications, and, in the MDR group, four patients (6%) did. Conclusions: There was no statistically significant difference in cause-specific survivals between the results of HDR and MDR brachytherapy for cervical cancer. The incidence of late complications tended to be higher for the HDR group than for the MDR group, but did not show a statistically significant difference (p=0.07).

During early pregnancy, the invasion of trophoblast cells into the decidua of the uterus is one of the essential steps in appropriate placentation. In this period, trophoblast cells are exposed to a relatively low-oxygen environment. The c-met protooncogene product ( Met), which is a high-affinity receptor for hepatocyte growth factor, plays an important role in controlling the invasion of many types of cells. The present study was designed to investigate the effect of low-oxygen tension on Met expression and the invasiveness of trophoblast cells isolated from early-stage human placenta and trophoblast-derived BeWo cells and JEG-3 cells. RT-PCR and immunoblot analyses demonstrated that low-oxygen tension (1% O-2) stimulated the expression of Met mRNA and protein, respectively. Hepatocyte growth factor production in the cells was not affected by oxygen tension. Transient transfection of BeWo cells with a hypoxia-inducible factor (HIF)-1 alpha expression vector to induce exogenous expression of HIF-1 alpha significantly increased the level of Met mRNA and protein, compared with transfection of a control vector. To examine whether this up-regulation of Met was directly induced by HIF-1 alpha, we performed the chromatin immunoprecipitation assay, which revealed that HIF-1 alpha binds to the promoter region of the Met gene under low-oxygen tension. JEG-3 cells cultured under 1% O-2 showed a more invasive character than those cultured under 20% O2, whereas inhibition of Met expression by small interfering RNAs prevented the low-oxygen, tension-induced invasiveness. These results suggest that the induction of Met expression by low- oxygen tension may play an important role in the physiology of early pregnancy by promoting the invasion of trophoblast cells into the decidua of the uterus.

Uterine artery embolisation (UAE) has become an alternative treatment for symptomatic uterine leiomyomata. Most reports suggest that it is well tolerated and effective, although there have been no reports of studies of biological parameters after UAE. In this study, we analysed the plasma level of vascular endothelial growth factor (VEGF) and the pulsatility index (PI) of uterine arteries before and after UAE. The level of plasma VEGF increased significantly after UAE (on day 1 and day 3) and decreased on day 7, and then increased again on day 30. The level of VEGF reached a peak value within three days after UAE. A significant inverse correlation was found between uterine artery PI and the level of VEGF on day 30, suggesting that VEGF may have negative effect on the efficacy of treatment of uterine leiomyomata by UAE.

During early pregnancy, trophoblast cells are exposed to relatively low-oxygen tension. Recently, the Rho GTPase family has been shown to play a key role in hypoxia-inducible factor-1 (HIF-1) alpha induction in renal cell carcinoma. The present study was designed to investigate the effect of low-oxygen conditions on RhoA expression in trophoblast cells isolated from early stages of human placenta and in trophoblast-derived BeWo cells and JAR cells. Immunoblot and RTPCR analyses showed that low-oxygen conditions (1% O-2 or 250 mu M CoCl2) stimulated expression of RhoA protein and mRNA. Pull-down assays demonstrated that these low-oxygen conditions increased RhoA activity. Preincubation of BeWo cells with Clostridium botulinum C3 exoenzyme, a specific inhibitor of Rho, inhibited hypoxia-induced HIF-1 alpha expression. Under 1% O-2 or 250 mu M CoCl2, BeWo cells, transfected with a dominant-negative RhoA, exhibited decreased levels of HIF-1 alpha protein and mRNA compared with the control vector transfectants. BeWo cells expressing constitutively active RhoA showed enhanced protein levels of not only HIF-1 alpha but also vascular endothelial growth factor ( VEGF) and glucose transporter 1, which are target gene products of HIF-1 alpha. These findings suggest that up-regulation of RhoA induced by low-oxygen conditions may play an important role in regulation of HIF-1 alpha expression in trophoblast cells.

細胞膜を通過する物質輸送には,担体(トランスポーター)を介し,濃度勾配に従ってエネルギーを必要としない促通拡散があり,担体を介さない単純拡散よりも早い速度で拡散される.また,この輸送とは異なりATPなどのエネルギーを使って濃度勾配に逆らって輸送される能動輸送がある.トロフォブラストでは,グルコーストランスポーター(GLUT)を介するグルコースの促通拡散と,アミノ酸トランスポーターを介するアミノ酸の能動輸送がよく知られている.今回,これらの担体を介する胎盤の物質輸送をわれわれのデータも含めて解説する.最後に,トロフォブラストの分化とGLUTの関連を説明する(著者抄録)

Neonatal necrotizing enterocolitis (NEC), which is a disease with a poor prognosis, is considered to be caused by the coincidence of intestinal ischemia-reperfusion injury and systemic inflammation due to the colonization of pathogenic bacteria. Interleukin (IL)-8, a proinflammatory cytokine, plays an important role in the pathophysiology of NEC. It was recently reported that IL-1beta activates the IL-8 gene by regulating the transcriptional nuclear factor kappaB (NF-kappaB) signaling pathways in intestinal cells. The protective role of maternal milk in NEC pathogenesis has been reported in both human and animal studies. In this study, we show that human breast milk dramatically suppressed the IL-1beta-induced activation of the IL-8 gene promoter by inhibiting the activation pathway of NF-kappaB. Moreover, we also show that human breast milk induced the production of IkappaBalpha. These results suggest that human breast milk could be protective and therapeutic in neonates with NEC by inhibiting the activation pathway of NF-kappaB.

Glucose transporter 1 (GLUT1) plays an important role in the transport of glucose in the placenta. During early pregnancy, placentation occurs in a relatively hypoxic environment that is essential for appropriate embryonic development, and GLUT1 expression is enhanced in response to oxygen deficiency in the placenta. Hypoxia-inducible factor-1 (HIF-1)alpha is involved in the induction of GLUT1 expression in other cells. The present study was designed to test whether HIF-1alpha is involved in hypoxia-induced activation of GLUT1 expression using trophoblast-derived human BeWo and rat Rcho-1 cells as models. GLUT1 mRNA and protein expression were elevated under 5% O-2 or in the presense of cobalt chloride, which has been shown to mimic hypoxia. Using rat GLUT1 (rGLUT1) promoter-luciferase constructs, we showed that this up-regulation was mediated at the transcriptional level. Deletion mutant analysis of the rGLUT1 promoter indicated that a 184 bp hypoxia-responsive element (HR-E) of the promoter was essential to increase GLUT1 reporter gene expression in response to low-oxygen conditions. BeWo and Rcho-1 cells cultured under 5% O-2 or with CoCl2 showed increased expression of HIF-1alpha protein compared with those cultured under 20% O-2. To test whether this factor is directly involved in hypoxia-induced GLUT1 promoter activation, BeWo and Rcho-1 cells were transiently transfected with an HIF-1alpha expression vector. Exogeneous HIF-1alpha markedly increased the GLUT1 promoter activity from constructs containing the HRE site, while the GLUT1 promoter constructs lacking the HRE site were not activated by exogenous HIF-1alpha. These data demonstrate that GLUT1 is up-regulated under 5% 02 or in the presence of CoCl2 in the placental cell lines through HIF-1alpha interaction with a consensus HRE site of the GLUT1 promoter.

Breast milk has non-nutritional protective effects on recipient infants. It has been speculated that bioactive substances present in human milk have important roles in protecting infants. However, the mechanisms by which such substances protect newborns are unclear. Therefore, we analyzed the growth-promoting activity of human milk and the intracellular signaling mechanism thereof using human fetal small intestinal (FHS 74 Int) cells. Epidermal growth factor (EGF) stimulated the proliferation of these cells. However, this stimulation was less effective than that of aqueous milk (5% vol/vol). The bioactivity of human milk was heat stable but protease sensitive. EGF receptor tyrosine kinase inhibitor did not repress the milk-induced growth-promoting effect on fetal small intestinal cells. Regarding the intracellular signaling pathway, the milk-induced cell proliferation pathway was tyrosine kinase dependent but was neither mitogen-activated protein (MAP) kinase nor phosphatidylinositol-3 (PI-3) kinase dependent. On the other hand, EGF-induced cell proliferation was tyrosine kinase, MAP kinase, and PI-3 kinase dependent. Rapid tyrosine phosphorylation of several intracellular proteins was detected after milk stimulation. Furthermore, the time course of phosphorylation induced by milk was different front that induced by EGF. The sizes of the proteins phosphorylated in response to milk were different from those of the Shc proteins phosphorylated in response to EGF. These results suggest that human milk induces fetal intestinal cell proliferation through a unique tyrosine kinase pathway different from the EGF receptor signaling pathway.

A case of fetal goitrous hypothyroidism associated with high-output cardiac failure is presented. At 32 weeks of gestation, the antenatal diagnosis of goiter was made based on ultrasound examination, and the fetal thyroid function was examined by amniocentesis and cordocentesis. Color and pulsed Doppler examinations demonstrated a high vascular flow Pattern in the goiter and marked elevation of the maximum velocity in the common carotid artery at the level of the neck. It was suspected that arteriovenous shunting through the large goiter resulted in high-output cardiac failure with cardiomegaly and pleural effusion. The fetus was treated by injection of levothyroxine sodium into the amniotic fluid at 33 weeks of gestation and the goiter thereafter decreased in size, with subsequent improvement of the high-output cardiac failure. The maximum velocity in the common carotid artery fell rapidly before the shrinkage of the fetal goiter and in parallel with the fetal level of thyroid-stimulating hormone.

Twin-to-twin transfusion syndrome (TTTS) is a severe complication of monochorionic twin pregnancies, with high perinatal loss and significant morbidity. The etiology of this syndrome remains unclear. Hyperreactio luteinalis is a rare benign condition characterized by maternal ovarian enlargement due to theca lutein cysts. We present 4 cases of hyperreactio luteinalis associated with severe TTTS. We detected maternal ovarian enlargement by prenatal ultrasonography in 2 cases, at the cesarean section in 1 case, and postpartum in 1 case. Three cases showed hydrops fetalis and all cases showed congestive cardiac failure. Serial amnioreduction was performed in all cases, and regression of hydrops fetalis and maternal ovarian enlargement occurred in 1 case. These cases suggest that the pathophysiology of TTTS is closely related to the etiology of hyperreactio luteinalis. Copyright (C) 2002 S. Karger AG, Basel.

Estrogen exerts a variety of effects not only on female reproductive organs but also on nonreproductive organs, including adipose tissue. Estrogen inhibits obesity triggered by ovariectomy in rodents. We studied the mechanism underlying this estrogen-dependent inhibition of obesity. Estrogen markedly decreased the amounts of fat accumulation and lipoprotein lipase (LPL) mRNA as well as triglyceride accumulation in genetically manipulated 3T3-L1 adipocytes stably expressing the estrogen receptor (ER). A pLPL(1980)-CAT construct, along with an ER expression vector, was introduced into differentiated 3T3-L1 cells, and CAT activities were determined. ER, mostly ligand-dependently, inhibited the basal LPL promoter activity by 7-fold. We searched the LPL promoter for an estrogen-responsive suppressive element by employing a set of 5'-deletion mutants of the pLPL-CAT reporter. Although there was no classical estrogen response element, it was demonstrated that an AP-1-like TGAATTC sequence located at (-1856/-1850) was responsible for the suppression of the LPL gene transcription by estrogen. An electrophoretic mobility shift assay probed with the TGAATTC sequence demonstrated formation of a specific DNA-nuclear protein complex. Interestingly, this complex was not affected by the addition of any antibodies against ER, c-Jun, c-Fos, JunB, or JunD. Because this TGAATTC element responded to phorbol ester and overexpression of CREB-binding protein abrogated the suppressive effect of estrogen on the LPL promoter, we conclude that a unique protein that is related to the AP-1 transcription factor families may be involved in the complex that binds to the TGAATTC element.

Leptin is the product of the obese gene (ob), and is secreted in plasma from mature adipocytes. It has been recently reported that leptin is synthesized in granulosa and cumulus cells within the follicle of the ovary, and is present in mature human oocytes, suggesting possible roles of leptin in several aspects of pre- and post-ovulatory follicular development. On the other hand, STAT (Signal Transducer and Activator of Transcription) transcription factors are involved in leptin-associated signal transduction. In this report, we studied the expression of leptin receptor and STAT3 activation by leptin in metaphase 2 stage (M2) oocytes. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting showed that mRNA and protein of leptin receptor were expressed in M2 stage oocyte. Leptin at 15 ng/ml, the concentration observed in follicular fluid, caused tyrosine phosphorylation of STAT3 in mouse M2 stage oocytes. These results suggest possible roles of leptin in several aspects during oocyte maturation by activating the STAT signal transduction pathway.

The regulation of mitogen-activated protein (MAP) kinase by endothelin-1 (ET-1) in cultured rat puerperal uterine myometrial cells was investigated. ET-1 caused the rapid stimulation of MAP kinase activity. ET-1-induced MAP kinase activation is neither extracellular Ca2+- nor intracellular Ca2+-dependent. ET-1 stimulation also led to an increase in phosphorylation of son-of-sevenless (SOS), and transfection of dominant negative SOS attenuated the ET-1-induced MAP kinase activity. Phorbol-12-myristate 13-acetate (PMA) also induced the MAP kinase activity, but pretreatment of the cultured cells with PMA, to down-regulate protein kinase C (PKC), did not abolish the activation of MAP kinase by ET-1. In addition, down-regulation of PKC had no effect on ET-1-induced SOS phosphorylation. Pertussis toxin, which inactivates Gi/Go proteins, blocked the ET-1-induced MAP kinase activation but not the PMA-induced MAP kinase activation. The results suggested that MAP kinase is acutely activated by ET-1 through a pertussis toxin-sensitive G protein and SOS, not through the PMA-sensitive PKC. In addition, although reverse-transcriptase PCR assays detected messenger RNA for both ET- 1 receptor subtypes in cultured rat puerperal uterine myometrial cells, ET-1-induced MAP kinase activity and uterine contraction were blocked by treatment with BQ485, an antagonist selective for an ET type A receptor (but not by BQ788, an ET type B receptor antagonist). Ritodrine, which is known to relax uterine muscle contraction, attenuated ET-1-induced MAP kinase activity. We further examined the role of MAP kinase pathway in uterine contraction using an inhibitor of MEK activity, PD098059. This inhibitor completely inhibited the ET-1-induced MAP kinase activation and partially, but significantly, inhibited the ET-1-induced uterine contraction. These results indicate that ET-1-induced MAP kinase signaling cascade may play an important role in the ET-1-induced uterine contraction.

Cytokines and steroid hormones use different sets of signal transduction pathways, which seem to be unrelated. Interleukin-6 (IL-6) uses JAK tyrosine kinase and STAT (signal transducer and activator of transcription) transcription factor. Glucocorticoid binds glucocorticoid receptor (GR), which is a member of the steroid receptor superfamily. We have studied the crosstalk between the IL-6-JAK-STAT and glucocorticoid-nuclear receptor pathways. IL-6 and glucocorticoid synergistically activated the IL-6 response element on the rat alpha2-macroglobulin promoter (APRE)-driven luciferase gene. The exogenous expression of GR enhanced the synergism. The exogenous expression of dominant negative STAT3 completely abolished the IL-6 plus glucocorticoid-induced activation of the APRE-luciferase gene. Tyrosine phosphorylation of STAT3 stimulated by IL-6 alone was not different from that by IL-6 plus glucocorticoid. The protein level of STAT3 was also not increased by glucocorticoid stimulation. The time course of STAT3 tyrosine phosphorylation by IL-6 plus glucocorticoid was not different from that by IL-6 alone. The synergism was studied on the two other IL-6 response elements, the junB promoter (JRE-IL-6) and the interferon regulatory factor-1 (IRF-1) promoter (IRF-GAS) which could be activated by STAT3. The synergistic activation by glucocorticoid on the IL-6-activated JRE-IL-6 and the IRF-GAS-driven luciferase gene was not detected. Glucocorticoid did not change the mobility of IL-6-induced APRE-binding proteins in a gel shift assay. These results suggest that the synergism was through the GR and STAT3, and the coactivation pathway which was specific for APRE was the target of glucocorticoid.

The purpose of this study was to analyze the mechanism of transcriptional activation of human chorionic gonadotropin-alpha (hCGalpha) gene by epidermal growth factor (EGF) in trophoblast cells. We stably transfected hCGalpha promoter-chloramphenicol acetyltransferase constructs into Rcho-1 trophoblast cells and monitored the promoter activities. -290-base pair hCGalpha promoter containing a tandem repeat of cAMP response element (CRE) was activated by EGF in a dose- and time-dependent manner. Deletion analysis of hCGalpha promoter suggested an involvement of CRE in EGF-induced hCGalpha transcriptional activation. Moreover, the hCGalpha promoter, of which both CREs were mutated, did not respond to EGF. These results indicate that EGF activates the hCGalpha gene transcription through CRE. Although EGF did not alter the amount of CRE-binding protein (CREB), EGF induced CREB phosphorylation. We next examined the mechanism of CREB phosphorylation by EGF. Protein kinase C inhibitors (H7, staurosporin, and chelerythrine) inhibited EGF-induced CREB phosphorylation, whereas either mitogen-activated protein kinase kinase-1 inhibitor (PD98059) or protein kinase A inhibitor (H8) showed no effect. Furthermore, H7 and staurosporin but not H8 inhibited hCGalpha promoter activation by EGF. In conclusion, EGF promotes hCGalpha gene transcription via the CRE region probably by phosphorylating CREB mainly through the protein kinase C pathway in trophoblast cells.

Leukemia inhibitory factor (LIF) is a cytokine that plays an important role during mouse embryogenesis. We showed that adenovirus E1A represses the interleukin-6 signal transduction pathway that uses the same JAK tyrosine kinase and STAT (signal transducer and activator of transcription) transcription factor as LIF. Here, we report that the LIF-JAK-STAT signal transduction pathway is blocked in cellular E1A-expressing undifferentiated F9 cells, and that the block is overcome by retinoic acid-induced differentiation. LIF failed to stimulate the expression of the acute phase response element (APRE)-driven luciferase gene in undifferentiated F9 cells, whereas the luciferase activity was remarkably increased by LIF treatment in differentiated F9 (dF9) cells. We analyzed the mechanism of the APRE regulation and found that the LIF-induced APRE-binding activity was regulated in a differentiation-dependent manner. The protein levels and the tyrosine phosphorylation of JAK1, JAK2, and STAT3 in F9 cells were not different from those in dF9 cells. The exogenous expression of activated c-Ha-ras partially recovered the LIF responsiveness of the APRE-luciferase gene in F9 cells, but the dominant negative ras N-17 did not repress the LIF-induced activation of APRE-luciferase in dF9 cells. These results suggested that an unknown coactivation process that is partially compensated by Ras is required for STAT3-APRE binding in F9 cells.

The signal transduction mechanism involved in human placental lactogen (hPL) was studied. We have identified that hPL rapidly stimulated the tyrosine phosphorylation of at least 7 proteins including Janus Kinases (JAK1 and JAK2) and a signal transducer and activator of transcription protein (Stat3). This is the first evidence that the JAK-STAT pathway is involved in the hPL signaling. Moreover, two unknown proteins which were different from STAT proteins (Stat1, 3 and 5) in sizes were predominantly tyrosine-phosphorylated. Because human growth hormone (hGH) activates Stat1, 3, 5 and human prolactin (hPRL) activates Stat5, these results show that hPL uses a unique signal transduction pathway which is different from hGH and hPRL.

BACKGROUND: The regulatory mechanism of tumor markers secretion has not been well clarified. METHODS: Serum levels of CA 125 and tissue polypeptide antigen (TPA) from 17 patients with Stage III serous cystadenocarcinoma were measured prior to an initial surgical treatment. Epidermal growth factor receptor (EGFR) status was examined by an 125I-EGF binding assay in a human serous cystadenocarcinoma cell (SHIN-3) and in the 17 primary carcinomas. SHIN-3 cell and the EGFR-expressing primary cancer cells (n = 4) were cultured with or without various concentrations of transforming growth factor (TGF-alpha), a ligand for EGFR, and the CA 125 and TPA concentrations in the conditioned media were measured. RESULTS: EGFR was expressed in 12 primary carcinomas and in the SHIN-3 cell, and it was absent in the remaining 5 carcinomas. Pre-therapeutic serum CA 125 and TPA levels were significantly greater (P < 0.05) in patients with EGFR-expressing carcinomas (n = 5). These data suggest a possible involvement of EGFR in regulating these tumor markers secretion. TGF-alpha increased the CA 125 and TPA secretion from SHIN-3 cell. It also promoted the CA 125 secretion in 2 of 4 EGFR-expressing primary ovarian carcinoma specimens. CONCLUSIONS: These results suggest that a signal through the EGFR may be involved in regulating the CA 125 and TPA secretion from human ovarian carcinomas.

Some viral products interfere with host antiviral defense mechanisms. Adenovirus E1A represses IFN signal transduction pathways which induces gene activation and an antiviral state. Both IFN and IL-6 activate Jak/Tyk protein tyrosine kinases and the STAT (signal transducer and activator of transcription) family proteins. We showed that 12S E1A repressed IL-6 signals activating the junB promoter and the two IL-6 response elements (REs), JRE-IL6 and type II IL-6 RE (also called acute phase response element), required for IL-6-induced activation of the junB promoter and the type II acute phase reactant genes, respectively, in hepatocytes. Conserved region 1 of the 12S E1A was responsible for the repression. Target molecules of the repression by E1A appeared to be IL-6-inducible DNA-binding proteins acting on the IL-6 REs. In a rat 3Y1 cell line stably expressing E1A, the levels of IL-6-induced IL-6 RE binding proteins were severely reduced compared with those in a parental 3Y1 cell line. Moreover, we found that the levels of the STAT family proteins including Stat1-alpha (p91), Stat1-beta (p84), Stat2 (p113), and Stat3 were decreased by the stable expression of adenovirus E1A. The E1A-induced reduction in the amount of DNA-binding proteins seemed to be partly responsible for the decreased transcriptional activity of the IL-6 RE-driven gene expression in response to IL-6. This repression mechanism may be applicable to the E1A repression of IFN-gamma-induced gene activation.

IL-6 signals activate an IL-6 response element in the junB promoter, JRE-IL6, in a Ras-independent manner. IL-6 rapidly induced a DNA-binding activity to the Ets binding site of JRE-IL6 (JEBS), one of necessary DNA motifs of the IL-6 response element. The IL-6-induced JEBS-binding activity was indistinguishable from those to acute phase response element (APRE) in both kinetics of induction and its DNA-binding specificity. Purified APRE binding factors (APRFs) from IL-6-stimulated rat liver were found to be composed of multiple Stat3-related proteins and Stat1. Moreover the purified APRFs specifically made a complex with JRE-IL6 with the same mobility as that observed in the crude extracts. These results indicate that an immediate early signal of IL-6 leading to activation of JRE-IL6 is mediated by STAT family transcription factors.

Interleukin-6 (IL-6) activation of the immediate-early gene junB has been shown to require both a tyrosine kinase and an unknown 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7)-sensitive pathway. Here we report the identification and characterization of an IL-6 immediate-early response element in the junB promoter (designated JRE-IL6) in HepG2 cells. The JRE-IL6 element, located at -149 to -124, contains two DNA motifs, an Ets-binding site (EBS) (CAGGAAGC) and a CRE-like site (TGACGCGA). Functional studies using variously mutated JRE-IL6 elements showed that both motifs were necessary and sufficient for IL-6 response of the promoter. The EBS of the JRE-IL6 element (JEBS) appears to bind a protein in the Ets family or a related protein which could also form a major complex with the EBSs of the murine sarcoma virus long terminal repeat or human T-cell leukemia virus type 1 long terminal repeat. The CRE-like site appears to weakly bind multiple CREB-ATF family proteins. Despite the similarity in the structure between the JRE-IL6 element and the polyomavirus enhancer PyPEA3, composed of an EBS and an AP1-binding site and known to be activated by a variety of oncogene signals, JRE-IL6 could not be activated by activated Ha-Ras, Raf-1, or 12-O-tetradecanoylphorbol-13-acetate. We show that IL-6 activates JRE-IL6 through an H7-sensitive pathway that does not involve protein kinase C, cyclic AMP-dependent kinase, Ca(2+)- or calmodulin-dependent kinases, Ras, Raf-1, or NF-IL6 (C/EBP beta). The combination of JEBS and the CRE-like site appears to form the basis for the selective and efficient response of JRE-IL6 to IL-6 signals, but not to signals generated by activated Ha-Ras, Raf-1, or protein kinase C.

月経前症候群（PMS）患者における腸内細菌叢の特徴を検討した。PMS症状を訴える女性27名と、症状が重度ではない女性29名を募集した。その中から、社会生活に支障をきたすほどの症状を持つ21名（PMD群）と、深刻な症状がなく、社会生活にも支障がない女性22名（対照群）をさらに選出した。血液中の炎症マーカーは、PMD群と対照群で有意差はなかった。PMD群と対照群の腸内細菌叢には、β多様性の差が検出された。属レベルでの腸内細菌の検討では、対照群との比較で、ParabacteroidesとMegasphaeraの減少がPMS患者の特徴となり、さらにこれらの量が少ないと、症状が重度となることが判明した。

月経困難症や月経前症候群などの月経関連疾患は、女性のQOLを著しく低下するものであり、近年でも増加している。また、月経は妊娠分娩と密接な関係がある。母子コホート調査であるエコチル調査より宮城県の妊婦2,310人を対象に、分娩後に再開した月経における疼痛や月経前の精神的不安定さの程度について検証した。 分娩後1.5年までに再開した月経において、月経痛の程度は、妊娠前の属性、妊娠中の不安や抑うつ、産後うつ、月経再開後の周期不順と関連し、月経前の精神的不安定さは妊娠中の喫煙や精神状態と関連していた。しかし月経困難症も月経前症候群も、周産期の合併症や異常経過とは関連していなかった。

大学生対象の調査結果から、アスリートのほうが、文化部学生と比較し、PMS/PMDDによるパフォーマンス障害が大きく、食事摂取面では魚摂取がパフォーマンス障害の改善因子となった（オッス比0.61）。より詳細な食事摂取調査票のBDHQを用いた調査結果からは、パフォーマンス障害を自覚する選手では、植物性蛋白摂取が少なく、動物性蛋白摂取が多くなった。活性型イソフラボンであるエクオール産生能を検討したところ、エクオール非産生者はパフォーマンス障害のリスク因子となった（オッス比3.34）。女子高校生運動部員を対象とする調査から、PMSの身体症状が疲労骨折のリスクファクターとなった（オッス比1.66）。

糖尿病治療薬であるメトホルミンを用いて、子宮筋腫細胞におけるAMPK・mTORシグナルを標的とした、細胞増殖抑制機構とVEGF産生抑制機構を明らかにした。メトホルミンはAMPKを活性化し、mTORシグナル伝達経路を抑制し、細胞増殖とVEGF産生の両者を抑制した。また、メトホルミンは低酸素下でのHIF-1をpost-transalational levelで抑制した。メトホルミンは細胞増殖と血管新生の両面から、子宮筋腫増大に抑制的に作用する可能性が示唆される結果が得られた。

メタボリックシンドローム(MS)で重要な役割をはたすアンジオテンシン・アルドステロンの筋腫細胞増殖機構を明らかにした。MSに抑制的に作用するアディポネクチンの筋腫細胞増殖抑制機構を明らかにした。植物性エストロゲンのジェニスタイン、ターメリック・ウコン中有効成分のクルクミンによる増殖抑制効果を検討し、PPARγを介する抑制系を明らかにした。子宮筋腫動物実験系として、ヒト化マウスを用いたヒト子宮筋腫移植モデルマウスを開発した。

最初にId-1遺伝子プロモーター(-1472bp)ルシフェレース遺伝子をRcho-1細胞に遺伝子導入し、正酸素下、低酸素下での未分化、分化による活性変化をルシフェレースアッセイを用いて検討した。正酸素下ではこの領域で分化に伴う転写活性の抑制が認められ、低酸素下では抑制が認められなかった。次に様々な長さのdeletion mutantを作成し、未分化状態における活性化の責任領域を検討した。-272bp〜-146bpに活性化部位が存在することが明らかとなった。この領域には、様々な転写因子の結合領域が存在する3領域(box1,2,3)が存在し、これらをもとにさらに短いdeletion mutantを作成したところ3つの領域すべてが、転写活性に関与することが明らかとなった。そこで3つの領域を1つずつ欠損したmutant(δ1,2,3)を作成し、どの領域が特に重要であるかを検討したところbox2領域(-203bp〜-179bp)が転写活性に特に重要であることが明らかとなった。box2には転写因子であるSP-1とNF-1の結合領域が存在しており、これらの因子が分化に伴うId-1遺伝子の転写制御に中心的役割をはたす可能性が考えられた。さらに分化した細胞にSP-1を過剰発現されると、Id-1遺伝子プロモーターは活性化され、SP-1がこのプロモーターの活性化に関与しうることが明らかとなった。現在、DAPAおよびEMSAを用いて、box2領域に結合する転写因子の解析・同定を試みている(以上の研究成果は、第76回日本内分泌学会総会、第11回日本胎盤学会総会、第55回日本産婦人科学会総会で発表)。なお、本年度購入予定であった3成分(N2-O2-CO2)インキュベーターは、医学部共同研究施設に代用可能な機種が導入されたため購入せず、その費用は薬品等の消耗物品購入に充当した。

蛋白質をはじめとする生体内機能分子の働きを生きた細胞内で観測する手段として、蛍光蛋白質を利用したFRET(Fluorescence energy transfer、蛍光エネルギー移動)を用いたエキソサイトーシス時のリガンド、受容体の細胞内移動動態を観察するために設定したFRETを用いたシステムの立ち上げに困難さを経験している。そこで、まず、いままでに実績のある、細胞内メッセンジャータンパク質である低分子量G蛋白のRasとRhoとおよびそのエフェクター蛋白であるRaf, PKNとの間のFRET現象の観察を試みた。細胞は我々にとって使い慣れており、かつ管理および操作が容易なCaov3(ヒト卵巣癌細胞由来細胞株)を用いた。RASにYFP(黄色蛍光色素)、RafにCFP(青色蛍光色素)を含むプラスミドを作成した。両者を一時的に発現することには成功したが、その安定的観察のためにはなんらかの科学的捜査が必要である。またFRET現象を観察するためには2分子の安定的バランスが必要である。両蛋白を同時に等量づつ発現させるために2つのプラスミドをhinge(蝶番)でつないで一つのプラスミドとして均等に発現するようシステムを開発中であるが、未だ確立に至っていない。平行してRhoにCFP、PKNにYFPをつないだプラスミドを作成し、検討も行っているが同様の問題を解決できないでいる。以上の困難さより計画自身の遂行にも困難さを来している。

ES細胞の自己複製に、転写因子であるOct3/4が重要である(Nature genetics 25:372-376,2000)。すなわちOct3/4の発現レベルが上昇すると、ES細胞は原始内胚葉へ分化し、Oct3/4の発現レベルが一定であると、ES細胞は未分化状態を維持する。逆に、Oct3/4の発現レベルが下降すると、ES細胞は胎盤になる栄養外胚葉に分化し、転写因子であるCdx2,Hand1の発現が誘導される。また、Cdx2の下流にあると考えられる転写因子Id-1が、絨毛細胞(trophoblast)のstem cellとしての維持に関与していることが示唆される。 我々は、Id-1の上流域を含むreporter遺伝子を用い、絨毛癌細胞の未分化状態と分化状態での転写制御機構を検討したところ、Id-1遺伝子プロモーター上の-267bpから-146bpに未分化状態における転写活性部位を認めた。第12回胎盤学会学術集会で、ワークショップとして報告し、論文投稿準備中である。 また、低酸素状態で誘導されるhypoxia inducible factor(HIF)-・が絨毛細胞の分化誘導に関与ることが指摘されている(J.Clin.Invest.105:577-587,2000)。低酸素が絨毛細胞の分化機構及ぼす影響を検討し、HIF-・によって分化マーカーであるGLUT1が誘導されることを見いだし、Endocrinol 183 145-154 2004)に発表した。さらに、我々は絨毛癌細胞において、低酸素負荷てHIF-1・の誘導は、低分子量蛋白G-proteinのRhoAの発現増加および活性化を介すること示し、(J Clin.Endocrinol.Metab.in press)に発表した。これが、妊娠中の絨毛細胞におけるHIF-1・の現増加の新たなメカニズムとなる可能性が示唆された。 bone morphogenetic protein 4(BMP4)がES細胞からTS(trophoblst stem)細胞への分化誘導因である可能性が示唆された(Nat.Biotechnol.12:1261-4,2002)。絨毛細胞の分化機構のBMP4の加より絨毛癌細胞において、絨毛性ゴナドトロピン(hCG)の分泌が軽度増加することが示された。

グリーンマウスを作成し卵と母体細胞の相互関係を研究すべく本研究を立ち上げた。その過程でトランスジェニックマウスに関しては当大学の他の研究室より供与が可能になった。そこで卵細胞と幹細胞の共培養による立体培養に関する研究を行った。最初系の確立を目指して腫瘍細胞を用いて試行した。さらにマトリゲルを用いた浸潤の評価も併せて行い得る系を確立した。その方法を工夫して立体培養を試みたがマウスの卵の性格上立体的に胎盤を較正する条件がなかなかととのわなかった。その間、卵の培養では発生過程において低分子量G蛋白RhoのインヒビターであるY27632は卵の胚胞形成過程で停止をおこすことが明らかになり、その過程は可逆的であることを明らかにした。またそのメカニズムにはNa+/K+ exchangerが関与していることも明らかになった。一方絨毛細胞の分化において絨毛細胞は通常より低酸素濃度下でGLUT1およびVEGFはアップレギュレートされ、また間にHIF-1α分化誘導が介在していることが明らかとなった。一方、癌細胞の浸潤に関しては、細胞運動、接着、浸潤に関しては低分子量G蛋白Rhoを介した系が活性化され、細胞骨格の変化を介して起こっていることが明らかとなり、アレンドロネートが浸潤抑制作用を有することを細胞レベルとLPAを用いた卵細胞癌腹膜播種モデルをもちいた生体レベルで証明した。このように卵発生過程からの研究は困難であったが、研究テーマを共通する関連の研究は飛躍的に進んだ。グリーンマウス細胞は絨毛細胞も得られるのでそこから研究が可能となってきた。

私たちは、これまで子宮筋の収縮機構における低分子量G蛋白RhoA/Rho-kinase経路の関与について基礎的な検討をしてきた。これまでの解析により以下のことを新たに明らかにした。(1)Lipopolysaccaride(LPS)による早産マウスのモデルを用い、in vivoにおいてRho-kinase阻害剤が早産を抑制した。(2)切迫早産の治療において子宮収縮抑制剤として使われているb2-agonisitであるritodrineの作用機構の検討で、ritodrineが子宮筋においてRhoの活性化を抑制していることを明らかにした。(3)LPSによる早産マウスのモデルにおいて、頚管成熟軟化における頚管の組織学的変化を検討したところ、LPSによって好中球、マクロファージの頚管内への浸潤が認められたが、Rho-kinase阻害剤はこれらの浸潤を抑制した。 これらの成果は、平成15年の日本産科婦人科学会、日本内分泌学会、日本生殖内分泌学会にて発表した。また、それぞれの成果を現在論文投稿中である。 以上の結果から、羊膜絨毛膜炎による早産において、RhoA/Rho-kinase経路の情報伝達遮断が、子宮筋の収縮抑制作用や、炎症細胞の浸潤抑制を介しての抗炎症作用により、新しい子宮収縮抑制の治療戦略となりうる可能性を示すものである。また、頚管成熟軟化における好中球、マクロファージの頚管内への浸潤の抑制は、Rho-kinase阻害剤が子宮頚管成熟の調節に応用できることも示唆している。現在は、さらに妊娠子宮の収縮制御機構を明らかにすることを目的に、RhoAおよびRho-kinaseの発現調節における性ステロイドホルモンの役割に焦点を当てて検討を進めているところである。

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