AIM: Premenstrual dysphoric disorder (PMDD) interferes with patients' work, school, social activities, or relationships with others. For female students with PMDD to be active in society, both the patients and the surrounding population should be familiar with PMDD, be positive for treatment, and be aware that its disruptions can negatively impact society. Hence, this study aimed to determine PMDD awareness and attitudes toward premenstrual symptoms among male and female university students in Japan and to investigate the probability of suspected PMDD. METHODS: Through a survey, we asked all participants about their awareness of PMDD and their attitudes toward premenstrual symptoms. Female participants were also screened for premenstrual symptoms using the Premenstrual Symptoms Questionnaire (PSQ). Those who met certain criteria were classified into the "PMDD" group. RESULTS: A total of 1846 students (949 females, 892 males, and 5 unspecified) completed the survey on awareness and attitude. Only 20.6% of them were familiar with PMDD. Some believed that premenstrual symptoms should be endured. Only 36.2% had the idea that the disruptions caused by these symptoms were a loss to society. The PSQ was completed by 920 females. We found that 3.7% were classified into the "PMDD" group. Some participants in the "PMDD" group were not familiar with PMDD, while some perceived that premenstrual symptoms should be endured. CONCLUSION: Despite having suspected PMDD, a certain percentage of women remain unaware of their condition and endure their symptoms. PMDD awareness through education is crucial to achieving an active life for females of menstruating age.

OBJECTIVES: Healthcare professionals are poorly educated about menopausal disorders. To address this issue, we launched an online Menopause Instructor Course, which is certified by the Japanese Society of Psychosomatic Obstetrics and Gynecology (JSPOG). METHODS: Courses covering general remarks, hormone replacement therapy (HRT), non-HRT treatments, mental health, decision-making, and healthcare support were held in 2023 and 2024. Cognitive behavioral therapy was included in 2024. JSPOG-member healthcare professionals who completed all on-demand lectures and passed a post-test received certification as Menopause Instructors. In 2024, a questionnaire survey assessed participant feedback and compared Obstetricians and Gynecologists (OB/GYNs) to other healthcare professionals. RESULTS: A total of 198 and 143 professionals were certified in 2023 and 2024, respectively, with most of them being OB/GYNs. Certified Menopause Instructors were located across Japan. A survey conducted in 2024 showed that 93.7% of respondents were satisfied with the course. OB/GYNs found general remarks, HRT, and non-HRT treatments to be significantly easier. The overall self-assessed understanding was high (8.04 ± 1.01), with OB/GYNs scoring significantly higher than other healthcare professionals (8.29 ± 0.95 vs. 7.87 ± 1.02, P < 0.05). CONCLUSIONS: The course successfully increased the number of trained professionals in menopause in Japan. Enhancing the course content to better serve OB/GYNs and other healthcare professionals is essential.

BACKGROUND: In the digital age, adolescents increasingly rely on online sources for health-related information. eHealth literacy-defined as the ability to find, evaluate, and apply online health information-plays a crucial role in health outcomes. However, limited research exists on eHealth literacy among Japanese high school students, particularly on its association with menstrual health and psychological well-being. OBJECTIVE: This study aimed to assess the eHealth literacy of Japanese female high school students and examine its association with premenstrual symptoms, psychological distress, loneliness, and self-esteem. METHODS: A cross-sectional, web-based survey was conducted in December 2024 among 1607 female students from 2 public high schools in Sendai, Japan. A total of 909 students with regular menstrual cycles completed all survey items. The survey included the eHealth Literacy Scale (eHEALS), Premenstrual Symptoms Questionnaire, Kessler Psychological Distress Scale (K6), Revised UCLA Loneliness Scale, Rosenberg Self-Esteem Scale, and a numerical rating scale for menstrual pain. Statistical analyses, including Student t tests, chi-square tests, correlation analyses, and logistic regression analyses, were used to examine the relationships between eHealth literacy and various health outcomes. RESULTS: The mean eHEALS score was 22.8 (SD 7.3), with 32.1% (292/909) of participants classified as having high eHealth literacy (eHEALS≥26). Students with higher eHealth literacy reported significantly lower levels of loneliness and higher self-esteem. The severity of premenstrual symptoms, particularly feeling overwhelmed, was significantly lower in the high eHealth literacy group. Additionally, interpersonal difficulties related to premenstrual symptoms were less prevalent among students with high eHealth literacy. Pearson correlation analysis indicated negative associations between the eHEALS score and psychological distress (K6) and loneliness, whereas a positive association was observed with self-esteem. Logistic regression analysis showed that high self-esteem was significantly associated with high eHealth literacy. CONCLUSIONS: This study highlights the importance of eHealth literacy in adolescent health care. Higher eHealth literacy is linked to lower levels of loneliness, higher self-esteem, and reduced premenstrual symptom severity, particularly feeling overwhelmed. Although the cross-sectional design limits causal conclusions, these findings suggest that higher eHealth literacy is associated with better mental and reproductive health in adolescents. Future research should adopt longitudinal designs, include more diverse populations-such as male adolescents-and explore additional contributing factors to better elucidate these associations.

Premenstrual syndrome (PMS) and Premenstrual Dysphoric Disorder (PMDD) significantly impair women's quality of life. A survey of obstetricians and gynecologists showed that 19.5% of the doctors preferred Kampo medicine, including Tokishakuyakusan (TSS), Kamishoyosan (KSS), Keishibukuryogan (KBG), and Yokukansan (YKS), as the first-choice treatment for these conditions. We aimed to analyze the characteristics of each Kampo prescription. A secondary analysis was conducted on survey results from members of the Japan Society of Obstetrics and Gynecology collected from September to November 2021. Data from 1,259 respondents treating PMS/PMDD were analyzed. Our correspondence analysis plotted relationships among treatments, showing that Kampo prescriptions of TSS, KSS, KBG, and YKS were distributed similarly to oral contraceptives (OCPs), but different from selective serotonin reuptake inhibitors (SSRIs), serotonin and norepinephrine reuptake inhibitors (SNRIs), and gonadotropin-releasing hormone analogs. Among the Kampo formulas, prescription YKS was the closest to SSRIs/SNRIs. Logistic regression analysis revealed that shorter physician experience (< 10 years, ≥ 10 and < 20 years) was associated with selecting prescriptions for TSS and KBG, whereas being a private practitioner was linked to selecting prescriptions for KSS and YKS (clinic vs. hospital OR: 1.57, 1.77; clinic vs. university OR: 1.53, 1.71). The prescription of YKS was also associated with choosing SSRIs/SNRIs (OR: 1.81), chasteberry (OR: 6.24), and other medications or supplements (OR: 2.41). Kampo prescriptions were strongly correlated with OCPs. Prescription TSS and KBG were likely chosen by less experienced practitioners, whereas prescription YKS was used by those more familiar with PMS/PMDD treatments, including SSRIs/SNRIs.

This study investigated the cross-sectional association between major dietary patterns and premenstrual syndrome (PMS). Participants were 427 women aged 22-50 y who responded to a mail survey in 2022-2023. Dietary patterns were derived using principal component analysis based on consumption of 52 food and beverage items ascertained by a validated self-administered diet history questionnaire. PMS were assessed using the Premenstrual Symptoms Questionnaire. Logistic regression analysis was used to estimate odds ratios of PMS according to tertiles of dietary pattern scores. The prevalence of moderate to severe PMS was 9.6% (41 women). We identified four dietary patterns: vegetable, tomato and fish, Japanese, and alcohol dietary patterns. No dietary pattern was significantly associated with PMS. However, the odds ratios of PMS in the highest tertile of the vegetable dietary pattern (characterized by high intake of vegetables, mushrooms, potatoes, seaweeds, and chicken) tended to be lower compared to the lowest tertile. The multivariable-adjusted odds ratio of PMS for the highest versus lowest tertile of the vegetable dietary pattern score was 0.69 (95% confidence interval 0.30-1.59). None of the dietary patterns were appreciably associated with PMS. The finding of lower odds of PMS among women with a high score for the vegetable dietary pattern deserves further investigation.

Calcium and vitamin D have been suggested to be associated with the amelioration of symptoms of premenstrual syndrome (PMS). However, evidence for an association between the dietary intake of these nutrients and PMS is limited and inconsistent. We examined the cross-sectional association of calcium and vitamin D intake with PMS. Participants were 390 women aged 22-49 y who responded to a mail survey in 2022-2023. Dietary intake was assessed using a validated self-administered diet history questionnaire. PMS were assessed using the Premenstrual Symptoms Questionnaire. Logistic regression analysis was used to estimate odds ratios of PMS according to tertiles of calcium and vitamin D intake with adjustment for potential confounding variables. The prevalence of moderate to severe PMS was 10% (39 women). Neither calcium nor vitamin D intake was significantly associated with PMS. However, calcium intake was associated with a decreased prevalence of PMS, albeit without statistical significance, with multivariable-adjusted odds ratios (95% CI) for PMS in the lowest through highest tertiles of calcium intake of 1.00 (reference), 0.47 (0.18-1.25), and 0.27 (0.07-1.08), respectively (p for trend=0.06). The odds ratio of PMS was low in the highest tertile of vitamin D intake compared with the lowest, but without statistical significance (odds ratio 0.56, 95% CI 0.19-1.66). Our findings suggest that calcium and vitamin D intake was not appreciably associated with PMS. The suggestive inverse association between calcium intake and PMS requires further investigation.

BACKGROUND: Unintended pregnancy at higher risk of perinatal mood disorders; however, concurrent factors such as socioeconomic conditions may be more critical to mental health than pregnancy intention. Mental health risks among individuals undergoing fertility treatment are inconsistent. We investigated mental health risks during pregnancy and parenthood in parents who conceived unintentionally or through fertility treatment compared to those who conceived naturally and intentionally. METHODS: We conducted a web-based study with 10,000 adults ≥ 18 years old, either pregnant or with a child aged < 2 years. Male and female respondents weren't couples. We analyzed 1711 men and 7265 women, after filtering out invalid responses. We used a questionnaire including conception methods (e.g., naturally conceived intended/unintended pregnancies, fertility treatment such as scheduled intercourse or ovulation inducers [SI/OI], intrauterine insemination [IUI], and in-vitro fertilization or intracytoplasmic sperm injection [IVF/ICSI]) and mental health risks (e.g., psychological distress, chronic pain, death fantasies). Using a modified Poisson regression, we estimated relative risks (RR [CI]) for mental health risks compared to those with intended pregnancies. RESULTS: Unintended pregnancy showed higher mental health risks during pregnancy in both genders, with women having significantly higher psychological distress, chronic pain, and death fantasies (RR 1.63 [1.05-2.54], RR 1.63 [1.14-2.33], and RR 2.18 [1.50-3.18], respectively). Women's death fantasies risk remained high in parenthood: RR 1.40 (1.17-1.67). In relation to fertility treatments, men using SI/OI during their partner's pregnancy showed higher mental health risks, especially for chronic pain (RR 1.75 [1.01-3.05]). Men who underwent IUI showed higher mental health risks during parenthood, notably death fantasies (RR 2.41 [1.13-5.17]). Pregnant women using SI/OI experienced higher mental health risks, with a significant risk of chronic pain (RR 1.63 [1.14-2.33]). Pregnant women using IVF/ICSI had a significantly lower risk of chronic pain (RR 0.44 [0.22-0.87]), but women who used IVF/ICSI had a significantly higher risk of death fantasies during parenthood (RR 1.40 [1.04-1.88]). CONCLUSIONS: Mental health risks vary by parenting stage (pregnancy or early parenthood) and gender, especially for those who conceived unintentionally or through fertility treatment. Both stages require adaptable mental health support for all parents. TRIAL REGISTRATION: N/A (non-interventional study).

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.

OBJECTIVE: Kamishoyosan (KSS), a traditional Japanese Kampo medicine, is widely used to treat neuropsychiatric symptoms in perimenopausal and postmenopausal women. We aimed to elucidate the functional mechanisms underlying KSS-mediated reduction of stress response behaviors and neuropsychological symptoms in perimenopausal and postmenopausal women. METHODS: Female mice were bilaterally ovariectomized (OVX) at the age of 12 weeks and exposed to chronic water immersion and restraint stress for three weeks. Among them, mice in the OVX+stress+KSS group were fed chow containing KSS from one week before exposure to chronic stress until the end of the experiment. Firstly, we performed a marble burying test and measured serum corticosterone levels to assess irritability and stress conditions. Next, we examined whether KSS affects microRNA-18 (miR-18) and glucocorticoid receptor (GR) protein expression, as well as the basal dendritic spine morphology of pyramidal neurons in the medial prefrontal cortex (mPFC) of postmenopausal chronic stress-exposed mice. Analyzed data were expressed as mean ± standard deviation. Tukey's post hoc test, followed by analysis of variance (ANOVA), was used for among-group comparisons. RESULTS: KSS administration normalized chronic stress-induced unstable emotion-like behavior and upregulated plasma corticosterone levels. Furthermore, KSS ameliorated GR protein expression by downregulating miR-18 expression in the mPFC and recovered the immature morphological changes in spine formation of pyramidal neurons in the mPFC of OVX mice following chronic stress exposure. CONCLUSIONS: KSS administration in postmenopausal chronic stress-exposed mice exerted anti-stress effects and improved the basal dendritic spine morphology of pyramidal neurons by regulating miR-18 and glucocorticoid receptor expression in the mPFC.

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.

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.

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.

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.

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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