Dapagliflozin Ameliorates Ovulation Disorders via Attenuating Activated Microglia-Mediated Hypothalamic Inflammation in HFD-Fed Mice.

INTRODUCTION: Sodium-glucose cotransporter 2 inhibitors (SGLT2is) have shown neuroprotective effects in obese mice. However, whether SGLT2i can ameliorate high-fat diet (HFD)-related ovulation disorders remains unknown. The aim of this research was to investigate whether dapagliflozin improves HFD-induced ovulatory dysfunction by attenuating microglia-mediated hypothalamic inflammation. METHODS: C57BL/6J female mice fed HFD were treated with dapagliflozin (1 mg/kg) for 22 weeks. Plasma insulin, leptin, luteinizing hormone (LH), estradiol (E2), and IL-1ß levels were also tested. Microglial morphology, cell numbers, and SGLT2 expression were evaluated using immunofluorescence. The expression of IL-1ß, NLRP3, kisspeptin, gonadotropin-releasing hormone (GnRH), SGLT2, insulin, and leptin receptors in the hypothalamus was determined using immunohistochemical staining. We also examined the effects of dapagliflozin on glucose metabolism and the release of inflammatory factor in palmitic acid (PA)-treated HMC3 cells. RESULTS: As expected, dapagliflozin improved HFD-induced metabolic disturbances, peripheral versus central insulin and leptin resistance and also restored the regular estrous cycle. Furthermore, dapagliflozin blunted microglia activation, NLRP3 inflammasome priming, hypothalamic inflammation, and increased the expression of GnRH and kisspeptin at proestrus in the hypothalamus. Additionally, dapagliflozin markedly reduced IL-6 and NO release and fat accumulation, decreased lactic acid production and glucose consumption, and inhibited mammalian target of rapamycin (mTOR) and hexokinase 2 (HK2) expression in PA-treated HMC3 cells. These effects suggest that dapagliflozin reduced the mTOR/HK2-mediated aerobic glycolysis. CONCLUSIONS: Dapagliflozin improved HFD-related ovulation disorders by regulating glucose metabolism through mTOR/HK2 signaling and attenuating microglia-mediated hypothalamic inflammation. These results validate the novel role for the neuroprotection of SGLT2i in HFD-induced obesity and ovulation disorders.

Meta analysis of ovulation induction effect and pregnancy outcome of acupuncture & moxibustion combined with clomiphene in patients with polycystic ovary syndrome.

Objective: Using Mesh Meta Analysis to evaluate the efficacy of Acupuncture & Moxibustion, Clomiphene, Acupuncture & Moxibustion combined with Clomiphene for treating Polycystic Ovary Syndrome (PCOS), in order to provide evidence-based medical evidence for whether to recommend Acupuncture & Moxibustion or Combine western medicine to treat PCOS. Methods: Eight databases including The Cochrane Library, Pubmed, Embase, Web of Science, CNKI, Wanfang Date, VIP and CBM were searched by computer. The included research period is from the establishment of the database to May 2023, which concerned with randomized controlled trials involving Acupuncture & Moxibustion, Clomiphene, Acupuncture & Moxibustion combined with Clomiphene on ovulation induction and pregnancy outcome in patients with PCOS. The duration of the research paper is from 2016 to 2023.The inclusion criteria refer to the Rotterdam standards issued by the European Center for Human Reproduction and Embryology and the American Society of Reproductive Medicine in January 2003, or the Expert Consensus on the Diagnosis and Treatment of Polycystic Ovarian Syndrome by the Endocrinology Group of the Obstetrics and Gynecology Branch of the Chinese Medical Association. Simultaneously exclude related diseases, repetitive literature, as well as literature with incomplete abstract information and no original data provided. Two researchers independently screened the literature, extracted data, and evaluated the risk of bias included in the study, using Stata17.0 software for a mesh meta-analysis. Results: Six randomized controlled trials were included, covering 1410 PCOS patients. Three interventions included Acupuncture & Moxibustion, Clomiphene, Acupuncture & Moxibustion combined with Clomiphene. Mesh Meta Analysis showed that in terms of improving ovulation rate, there was no statistical difference between Acupuncture & Moxibustion (A), Clomiphene (B), Clomiphene combined with Acupuncture & Moxibustion (C) (P>0.05).Acupuncture & Moxibustion (A) versus Clomiphene (B) [MD=0.15,95% CI (-0.51,0.80)], Acupuncture & Moxibustion (A) versus Clomiphene combined with Acupuncture & Moxibustion (C) [MD=1.60,95% CI (0.97,2.23)], Clomiphene (B) versus Clomiphene combined with Acupuncture & Moxibustion (C) [MD=1.45,95% CI (0.91,1.99)]. In terms of pregnancy outcome, the difference between the three intervention methods was statistically significant (P<0.05). Acupuncture & Moxibustion (A) versus Clomiphene (B) [MD=-0.80,95% CI (-1.84,0.23)], Acupuncture & Moxibustion (A) versus Clomiphene combined with Acupuncture & Moxibustion (C) [MD=0.29,95% CI (-0.73,1.30)], and Clomiphene (B) versus Clomiphene combined with Acupuncture & Moxibustion (C) [MD=1.09,95% CI (0.39,1.79)], The order of pregnancy rate from high to low is Acupuncture & Moxibustion combined with Clomiphene (C), Acupuncture & Moxibustion (A), Clomiphene (C).In terms of influencing endometrial thickness, the difference between the three intervention methods was statistically significant (P<0.05). Acupuncture & Moxibustion (A) versus Clomiphene (B) [MD=-0.84,95% CI (-1.87,0.19)], Acupuncture & Moxibustion (A) versus Acupuncture & Moxibustion combined with Clomiphene (C) [MD=0.26,95% CI (-1.01,1.53)], Clomiphene (B) versus Acupuncture & Moxibustion combined with Clomiphene (C) [MD=1.10,95% CI (0.36,1.84)], Acupuncture & Moxibustion combined with Clomiphene (C) has the best effect on improving endometrial thickness. In subgroup analysis, the effect of Acupuncture & Moxibustion treatment frequency on ovulation rate and pregnancy rate was not statistically significant. The combination of Acupuncture & Moxibustion, Electroacupuncture and warm Acupuncture & Moxibustion has no effect on the pregnancy rate, but the combination of Electroacupuncture and Clomiphene has the best effect on improving the ovulation rate. In the observation of adverse reactions, compared with clomiphene alone, Acupuncture & Moxibustion combined with Clomiphene can reduce the occurrence of Luteinized Unruptured Follicle Syndrome (LUFS) and Ovarian Hyperstimulation Syndrome (OHSS), and reduce the occurrence of physical adverse reactions such as nausea, vomiting, headache and dermatitis. Conclusion: Acupuncture & Moxibustion is effective in improving the ovulation promoting effect and pregnancy outcome of PCOS patients. The ovulation promoting effect of Acupuncture & Moxibustion or combined with Clomiphene is similar to that of Clomiphene alone, but Acupuncture & Moxibustion combined with Clomiphene has more advantages in improving the pregnancy rate of PCOS, and it also can reduce the adverse reactions of Clomiphene alone. Acupuncture & Moxibustion can be used as a recommended treatment for PCOS. More cases should also be included in the subgroup analysis to study the impact of Acupuncture & Moxibustion programs on clinical efficacy and further optimize the Acupuncture & Moxibustion treatment program. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/#myprospero, identifier (CRD42023433057).

Effects of vitamin D supplementation on ovulation and pregnancy in women with polycystic ovary syndrome: a systematic review and meta-analysis.

Objective: To evaluate the effect of vitamin D supplementation on pregnancy and ovulation in patients with polycystic ovary syndrome. Method: We searched Pubmed, Medline (via Ovid, 1974 to 2020), EMBASE (via Ovid, 1974 to 2020), Cochrane Central Register of Controlled Trials (via Ovid), Web of Science, CNKI, WangFang and the Vip database from inception until April 2021. Two researchers independently screened articles, collected data and evaluated the quality, with Review manager 5.3 for meta-analysis. Results: Totally 20 randomized controlled studies with 1961 subjects were included. Meta analysis showed that pregnancy rate [RR=1.44 (1.28, 1.62), p<0.00,001], ovulation rate [RR=1.42 (1.14, 1.78), p=0.002] and matured oocytes rate [RR=1.08 (1.03, 1.13), p=0.002] of vitamin D supplementation group were significantly higher than those of control group. Meanwhile, early miscarriage rate [RR=0.44 (0.30, 0.66), p<0.00,001], androgen level [MD=-2.31 (-3.51, -1.11), p=0.0002], luteinizing hormone [MD=-1.47 (-2.57, -0.36), p=0.009], follicle stimulating hormone [MD=-0.15 (-0.24, -0.05), p=0.002], and premature delivery rate [RR=0.38, 95% CI (0.21, 0.70), p=0.002] were declined significantly than the controls. However, only one article suggested that the progesterone [MD=6.52 (4.52, 8.52), p<0.05] in the vitamin D intervention group was increased. There was no notable difference in the biochemical pregnancy rate [RR=0.95 (0.55, 1.63), p=0.84], gestational hypertension rate [RR=0.40, 95% CI (0.15, 1.11), p=0.08], gestational diabetes mellitus rate [RR=0.27, 95% CI (0.05, 1.39), p=0.11], fertilization rate [RR=1.05 (1.00, 1.10), p=0.04], cleavage rate [RR=1.03 (0.99, 1.06), p=0.17], high-quality embryo rate [RR=1.08 (0.98, 1.20), p=0.10], endometrial thickness [MD=0.10], 77 (-0.23, 1.77), p=0.13], estrogen level [MD=-0.34 (-1.55, 0.87), p=0.59], LH/FSH [MD=-0.14, 95% CI (-0.48, 0.20), p=1.00] and anti-Mullerian hormone [MD=-0.22 (-0.65, 0.21), p=0.32]. Conclusion: Vitamin D supplementation contribute to the higher pregnancy and ovulation rates, and lower androgen, LH, FSH and early miscarriage rates in women with PCOS, regardless of the use of ovulation induction drugs or assisted reproductive technologies. However, no significant improvement was observed in fertilization rate or cleavage rate. Due to the limitation in quality of involved studies, more high-quality RCTs are needed for further validation. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO, identifier CRD42021250284.

Mixed Method Evaluation of My Vital Cycles®: A Holistic School-Based Ovulatory Menstrual Health Literacy Program.

There is a high prevalence of ovulatory menstrual (OM) dysfunctions among adolescents, and their menstrual health literacy is poor. The OM cycle can be used as a personal health monitor provided that the skills to understand it are correctly taught. My Vital Cycles®, a holistic school-based OM health literacy program, was trialed with a Grade 9 cohort in one single-sex school in Western Australia using the Health Promoting School framework. A validated OM health literacy questionnaire was administered pre- and post-program with 94 participants. Functional OM health literacy improved overall, with 15 out of 20 items showing improvement post-program (p < 0.05). In addition, 19 out of 53 items for interactive OM health literacy, and 18 out of 25 items for critical OM health literacy improved (p < 0.05). The improvement in mood concerns (p = 0.002) was unexpected. Thematic analysis of three focus groups of 18 girls revealed four themes of increasing comfort levels; finding the program informative; inclusion of non-teaching support such as healthcare professionals; and suggestions for future refinements. Overall, this Western Australian PhD project which developed and trialed My Vital Cycles® improved OM health literacy and was positively received. Future research possibilities include understanding the program's impact on mental health and further trials in co-educational settings; amongst different populations; and with extended post-program testing.

Ovulation induction with letrozole and dexamethasone in infertile patients with letrozole-resistant polycystic ovary syndrome.

PURPOSE: To assess efficacy of adjuvant dexamethasone during letrozole cycles for ovulation induction (OI) in women with letrozole-resistant polycystic ovary syndrome (PCOS). METHODS: We retrospectively evaluated 42 cycles of OI from 28 infertile women with letrozole-resistant PCOS between September 2019 and November 2022. Letrozole was initiated on cycle day 3 for 5 days and increased via a stair-step approach to 7.5 mg as indicated. Patients were deemed letrozole-resistant if no dominant follicle was identified on transvaginal ultrasound following this dose. Resistant patients then received 5 additional days of letrozole 7.5 mg with low-dose dexamethasone 0.5 mg for 7 days and had a repeat ultrasound. The primary outcome was ovulation rate determined by the presence of a dominant follicle on ultrasound. Secondary outcomes included endometrial thickness, number of measurable follicles, and pregnancy outcomes among responders. RESULTS: Twenty-two of 28 (79%) letrozole-resistant PCOS patients had evidence of ovulation after the addition of dexamethasone in 35 out of 42 (83%) cycles. Clinical pregnancy occurred in 20% of ovulatory cycles with a cumulative rate of 32%. All clinical pregnancies resulted in a live birth. Patients who responded to adjuvant dexamethasone were more likely to have a shorter duration of infertility; however, there were no differences in other demographics, serum androgens including DHEA-S, or pretreatment glycemic status. CONCLUSION: Adding dexamethasone to letrozole increased ovulation rates in letrozole-resistant PCOS patients undergoing OI with similar pregnancy outcomes to prior studies. The addition of dexamethasone is an effective, inexpensive, and safe option for PCOS patients otherwise at risk for cycle cancelation.

Dietary n-3 PUFA augments pre-ovulatory follicle turnover and prolificacy in well-fed ewes.

The present study evaluated the effect of dietary supplementation with n-3 polyunsaturated fatty acids (PUFA) on preovulatory follicle (POF) turnover, prolificacy, and endocrine and metabolic milieu in Malpura sheep. Fifty cyclic ewes with 3-3.5 body condition scores on a five-point scale were allocated equally to two groups (n = 25) following estrus synchronization and were supplemented with 0.6 mL/kg body weight of n-3 PUFA-rich fish oil (FO) or palm oil (PO) as control, for 60 d following an acclimatization period of 7 d. All ewes were mated with sexually active rams at the end of the supplementation period. On ultrasonographic ovarian scanning at the last fourth estrus, the mean number of POFs was 77.8% greater (P < 0.01) in FO ewes than in the PO ewes. The proportion of ewes with multiple ovulations two months after the beginning of supplementation was 56% in the FO group as compared to 8% in the PO group. The number of fetuses was 46% higher (P < 0.01) in the FO than in the PO ewes at d 45 of gestation. At lambing, the twinning percent in the FO ewes was three times greater than in the PO ewes (27.3 vs. 9.1%). Plasma cholesterol, estradiol, and insulin concentrations were lower (P < 0.01) in ewes fed with FO than those offered PO group at the end of the feeding period. It was concluded that the dietary supplementation of n-3 PUFA-rich FO in well-fed Malpura ewes improved the number of follicles and ovulation rate which led to an increased prolificacy, accompanied by a reduction of plasma cholesterols, estradiol, and insulin.

Impact of progesterone-free luteal phase support following natural cycle frozen embryo transfer: Study protocol for a multicenter, non-inferiority, randomized controlled trial.

Introduction: Nowadays, frozen-thawed embryo transfer (FET) has become one of the standard treatments for infertility in the field of assisted reproductive technology (ART). Natural cycle FET (NC-FET) has many advantages, such as simplicity and economics, no effect on patients' menstrual cycles, estrogen and progesterone levels, as well as no interference in endometrial growth and transformation, which is aligned with the natural physiological state of embryo implantation. Nonetheless, there is a controversy regarding the need for luteal phase support (LPS) during NC-FET cycles. The purpose of this study is to assess whether LPS was not inferior to non-LPS in terms of OPR in NC-FET cycles. Methods and analysis: This study including 1,010 ovulatory women undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles with an elective freeze-all strategy followed by NC-FET will be performed at four university-affiliated reproductive centers. Participants will be randomly assigned in a 1:1 ratio to receive LPS treatment or not. This study is designed as an open-label, non-inferiority, randomized controlled trial (RCT), and the primary statistical strategies were intention-to-treat (ITT) and per-protocol (PP) analysis. Discussion: There may not have been any significant difference in the chance of a live birth after FET if no progesterone was supplemental during the luteal phase. However, due to the limited number of previous studies, which are mainly retrospective, evidence is still limited. Thus, by conducting this multicenter RCT, we intend to evaluate whether LPS is necessary in NC-FET. Ethics and dissemination: A Reproductive Ethics Committee of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine (SDUTCM) has approved this study. This study will handle the data as required by general data protection regulations. Participants will sign a written informed consent regarding participation in the study and storage of blood samples in a biobank for future research. This study will be monitored by study personnel trained in Good Clinical Practice who are not involved in the study. The results of this study will be disseminated through publication in international peer-reviewed scientific journals. Clinical trial registration: [https://www.chictr.org.cn/], identifier [ChiCTR2200057498].

Preconception vitamin D and miscarriage in a prospective cohort study.

STUDY QUESTION: Is preconception vitamin D level associated with the risk of miscarriage? SUMMARY ANSWER: Preconception vitamin D levels are not associated with the risk of miscarriage in a population of women conceiving naturally. WHAT IS KNOWN ALREADY: In humans, low vitamin D has been associated with prolonged menstrual cycles, delayed ovulation and a lower probability of conception. Animal and in vitro data indicate that vitamin D may affect implantation. STUDY DESIGN, SIZE, DURATION: This prospective time-to-pregnancy study included 362 women who were trying to conceive naturally between 2008 and 2015. PARTICIPANTS/MATERIALS, SETTING, METHODS: This study included participants who had been trying to conceive naturally for 3 months or less at enrollment and aged 30-44 years. A preconception blood sample was collected and 25-hydroxyvitamin D [25(OH)D] was measured. Women who conceived (N = 362) were at risk of a miscarriage from the day of a reported positive pregnancy test until either a participant-reported pregnancy loss or 20 weeks post day of last menstrual period, whichever came first. Gestational age was defined by ovulation. Time to miscarriage (days) or censoring was modeled using a multivariate Cox proportional hazards model. Multiple imputation was performed for missing covariates and missing day of ovulation. MAIN RESULTS AND THE ROLE OF CHANCE: The mean age was 33 years (SD: 3.0 years). Mean 25(OH)D was lower among those who reported their race as African-American and those with a higher BMI. After adjustment for age, race, BMI, education, exercise, alcohol and caffeine intake, compared to the referent group (30-<40 ng/ml), the hazard ratio (HR) and 95% CI for those with a low 25(OH)D level (<30 ng/ml) was 1.10 (CI: 0.62, 1.91). Among participants with a higher 25(OH)D level (≥40 ng/ml), the HR was 1.07 (CI: 0.62, 1.84). LIMITATIONS, REASONS FOR CAUTION: This study was limited by a 25(OH)D measurement at only a single time point. A large percentage of women in this study had sufficient vitamin D levels, which may have limited our power to detect an effect of deficiency. Women in this study were older (30-44 years), and predominantly reported their race as White which may limit generalizability. WIDER IMPLICATIONS OF THE FINDINGS: The findings of this study do not suggest an association between preconception vitamin D and miscarriage. Future research should focus on women at greater risk for miscarriage or in populations at risk for vitamin D deficiency or on supplementation. STUDY FUNDING/COMPETING INTEREST(S): This research was supported in part by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences (Z01ES103333). This research was also supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development of the National Institutes of Health (NIH) under award numbers R00HD079659 and R01HD067683. The authors have no conflicts of interest. TRIAL REGISTRATION NUMBER: N/A.

Combination of alpha lipoic acid and metformin supplement improve assisted reproductive technologies outcomes in polycystic ovary syndrome patients.

We aimed to investigating the effects of metformin (MET) in combination with alpha lipoic acid (ALA) on hormonal and biochemical parameters, in polycystic ovary syndrome (PCOS) women undergoing intracytoplasmic sperm injection (ICSI). This experimental pilot study with a randomized design was carried out on 40 PCOS women in two groups: (1) MET group, administered 1,500 mg/day MET, and (2) MET (1,500 mg/day)+ALA (1,800 mg/day) group. Drugs were administered from the third day of the previous cycle until the day of oocyte aspiration (six weeks of treatment in total). MET+ALA significantly increased the number of maturated oocytes and the rate of fertilization when compared to the MET group. Combination MET+ALA could increase significantly the number of oocytes retrieval and the number of good-quality embryos. Also, the malondialdehyde (MDA) level decreased significantly in the MET+ALA group and the total antioxidant capacity (TAC) level increased significantly in the MET+ALA group compared to the MET group. Also, fasting blood sugar (FBS), insulin, luteinizing hormone (LH), and LH/follicle stimulating hormone (FSH) levels were significantly lower in the MET+ALA group. The pregnancy outcomes showed no significant difference in the rates of biochemical pregnancy, clinical pregnancy, miscarriage, and live births between the control and study groups. The combination of MET+ALA treatment could moderate the complications of PCOS and subsequently improve oocyte and embryo quality.

Circadian Rhythms in the Neuronal Network Timing the Luteinizing Hormone Surge.

For billions of years before electric light was invented, life on Earth evolved under the pattern of light during the day and darkness during the night. Through evolution, nearly all organisms internalized the temporal rhythm of Earth's 24-hour rotation and evolved self-sustaining biological clocks with a ~24-hour rhythm. These internal rhythms are called circadian rhythms, and the molecular constituents that generate them are called molecular circadian clocks. Alignment of molecular clocks with the environmental light-dark rhythms optimizes physiology and behavior. This phenomenon is particularly true for reproductive function, in which seasonal breeders use day length information to time yearly changes in fertility. However, it is becoming increasingly clear that light-induced disruption of circadian rhythms can negatively impact fertility in nonseasonal breeders as well. In particular, the luteinizing hormone surge promoting ovulation is sensitive to circadian disruption. In this review, we will summarize our current understanding of the neuronal networks that underlie circadian rhythms and the luteinizing hormone surge.

Effects of Hochuekkito, a traditional Japanese medicine (Kampo), on reproduction of aging female mice.

PURPOSE: To determine the potentials of Hochuekkito (HET) treatment for aging infertility. METHODS: Mice at 36 weeks of age were fed without (control, n = 40) or with low (100 mg/kg/day, n = 24) and high (1000 mg/kg/day, n = 38) doses of HET for 12 weeks. Aging animals at 48 weeks of age were used for in vitro fertilization-embryo transfer (IVF-ET), and their ovaries were subjected to histological and quantitative inflammation analyses. RESULTS: HET administration decreased transcript levels of ovarian inflammatory markers, interleukin 6 (IL-6), IL-1ß, tumor necrosis factor (TNF)-α, and interferon-gamma (IFN-γ) but suppressed ovulation rates and the number of ovulated oocytes in aging mice. Furthermore, HET treatment decreased the rates of oocytes maturation and fertilization and the cumulus-cell expression of TNF-α-induced protein 6 and epidermal growth factor receptor. After IVF-ET, no improvement of declined live offspring rate by aging was achieved by HET administration, although there were no adverse effects on embryo development and implantation as well as gross morphology and bodyweight of pups. CONCLUSION: Present study indicated HET treatment interfered with ovulation and fertilization in aging mice without affecting ovarian follicle development. No improvement on the age-associated decline of live offspring rate and follicle development was achieved after HET treatment.

Acupuncture regulates the autophagy of ovarian granulosa cells in polycystic ovarian syndrome ovulation disorder by inhibiting the PI3K/AKT/mTOR pathway through LncMEG3.

The main features of polycystic ovary syndrome (PCOS) are abnormal follicular development and ovulation dysfunction, which are caused by the excessive autophagy of ovarian granulosa cells. Acupuncture has been shown to improve ovulation dysfunction and abnormal follicular development in PCOS patients, but its mechanism is unclear. This study hypothesized that the beneficial effects of acupuncture are the result of LncMEG3-mediated effects on the PI3K/AKT/mTOR pathway. Acupuncture (CV-4, RN-3, CV-6, SP-6 and EX-CA 1) was used to treat a rat model of polycystic ovary syndrome. Hematoxylin-eosin staining was used to observe ovarian morphology and enzyme-linked immunosorbent assay, western blotting, immunohistochemistry and real-time PCR were used to detect LH, E2, FSH, T, AMH, LncMEG3, PI3K, AKT, mTOR, P62 and LC3II/I expression. The ovarian morphology of 90% of the rats in the acupuncture treatment group was significantly improved after 11 consecutive days of therapy. Acupuncture also resulted in a significant decrease in serum LH, FSH, T and AMH levels and a significant increase in E2 level (P<0.01). LncMEG3, PI3K, AKT, mTOR, P62 and LC3II/I expression was decreased in ovarian granulosa cells after acupuncture compared with PCOS and lentiviral Intervention Group (P<0.05), while the expression of follicle stimulating hormone receptor was increased (P<0.05). These results indicate that acupuncture can down-regulate the expression of LncMEG3 and thereby inhibit the PI3K/AKT/mTOR pathway, reducing granulosa cell autophagy and normalizing their proliferation. These factors ultimately remedy abnormal follicular development. These findings suggest that acupuncture has clinical potential as a safe treatment for PCOS ovulatory dysfunction.

Dietary Supplements for Female Infertility: A Critical Review of Their Composition.

Infertility is the condition of about 15% of couples that cannot get a conception after one year of unprotected sexual intercourse. In females, the reduced reproductive capacity underlies the most varied causes. Dietary supplements (DS) might be used to improve the pregnancy rate and a wide range of DS are proposed today to support female fertility. Although many authors demonstrated the positive effect of some of these products, the real efficacy of this approach is still debated. In order to evaluate the potential efficacy of DS for female infertility, we analysed the products marketed in Italy, using an original approach. A review of literature was performed to evaluate the effect of nutraceuticals on various female reproductive outcomes and to detect the minimal effective daily dose (mED) able to improve at least one of these. Thereafter, we conceived a formula to classify the expected efficacy of each DS. Each DS was scored and included into three classes of expected efficacy: higher, lower, and none. Ten out of 24 supplements (41.7%) resulted in the higher and 8 (34.3%) in the lower efficacy group, the remaining 6 DS (25.0%) were expected to have no efficacy. DS marketed in Italy are usually blends of many substances that are frequently employed at a negligible dose or without any evidence of efficacy. These findings raise serious doubt about the potential effectiveness of most commercial DS for female infertility.

Berberine improves ovulation and endometrial receptivity in polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a clinical syndrome with reproductive and endocrine disorders. Berberine is a monomer from Chinese herbs such as Coptis chinensis, whose effect on improving ovulation and endometrial receptivity of PCOS is uncertain. PURPOSE: To evaluate the effect of berberine on improving PCOS and explore the mechanism. METHODS: The rat model of PCOS was induced by intraperitoneal injection of testosterone propionate. Then they was divided into model (Mod) group, low-dose of berberine (BL) group, high-dose of berberine (BH) group and metformin (Met) group as well as a control (Con) group was established. Ovary morphology, hormone level, glucolipid metabolism were measured. UID-mRNA-seq of ovary tissue was conducted to seek the mechanism of berberine on improving ovulation. Three biomarkers of endometrial receptivity were also examined in endometrium by immunohistochemistry. RESULTS: The number of cystic follicles was increased while the number of corpus luteum was decreased in the rats of Mod group. These changes could be reversed by high-dose of berberine intervention. Berberine could also decrease the levels of serum luteinizing hormone (LH) and total cholesterol (TC) in PCOS rats. Meanwhile, berberine improved the impairment of abnormal oral glucose tolerance without affecting fasting insulin level and Homeostasis model assessment-insulin resistance (HOMA-IR). Luteinizing hormone/ choriogonadotropin receptor (LHCGR) and cytochrome P450 Family 19 Subfamily A Member 1 (CYP19A1) were focused via RNA-seq of ovary. Protein expression in ovary and mRNA expression in granulosa cell of LHCGR and CYP19A1 were decreased in Mod group and rescued by the intervention of berberine. A decrease of endometrial thickness and an increase of integrin αvß3 and lysophosphatidic acid receptor 3 (LPAR3) protein expression were observed in Mod group, which could be also reversed by berberbine. CONCLUSIONS: Berberine could improve ovulation in PCOS and the mechanism might be associated with up-regulating LHCGR and CYP19A1. Berberine could also improve endometrial receptivity through down-regualting αvß3 and LPAR3.

The circadian system: From clocks to physiology.

The circadian system, composed of the central autonomous clock, the suprachiasmatic nucleus (SCN), and systems of the body that follow the signals of the SCN, continuously change the homeostatic set points of the body over the day-night cycle. Changes in the body's physiological state that do not agree with the time of the day feedback to the hypothalamus, and provide input to the SCN to adjust the condition, thus reaching another set point required by the changed conditions. This allows the adjustment of the set points to another level when environmental conditions change, which is thought to promote adaptation and survival. In fasting, the body temperature drops to a lower level only at the beginning of the sleep phase. Stressful conditions raise blood pressure relatively more during the active period than during the rest phase. Extensive, mostly reciprocal SCN interactions, with hypothalamic networks, induce these physiological adjustments by hormonal and autonomic control of the body's organs. More importantly, in addition to SCN's hormonal and autonomic influences, SCN induced behavior, such as rhythmic food intake, induces the oscillation of many genes in all tissues, including the so-called clock genes, which have an essential role as a transcriptional driving force for numerous cellular processes. Consequently, the light-dark cycle, the rhythm of the SCN, and the resulting rhythm in behavior need to be perfectly synchronized, especially where it involves synchronizing food intake with the activity phase. If these rhythms are not synchronous for extended periods of times, such as during shift work, light exposure at night, or frequent night eating, disease may develop. As such, our circadian system is a perfect illustration of how hypothalamic-driven processes depend on and interact with each other and need to be in seamless synchrony with the body's physiology.

[Mechanism of Ziyin recipe for treatment of ovulatory infertility: a network pharmacology-based study and clinical observations].

OBJECTIVE: To explore the mechanisms of Ziyin recipe for treatment of ovulatory infertility based on network pharmacology analysis and evaluate the clinical efficacy of this recipe. OBJECTIVE: TCMSP, PubChem, Genecards, String, Swiss Target Prediction, and Uniprot databases were searched to identify all the action targets of Ziyin recipe and ovulatory infertility to construct the PPI network. Gene ontology (GO) and KEGG pathway enrichment analyses were performed and the "TCM-active ingredient-target-pathway" network was constructed using Cytoscape 3.6.0. We also designed a controlled clinical trial to verify the clinical effectiveness of Ziyin recipe. The patients were randomized into 2 groups to receive treatment with Western medicine including CC and HMG (control group) and additional treatment with Ziyin recipe, and the dosage of GN, follicular development days, E2 level of a single dominant follicle on trigger day, ovulation rate, the rate of LUFS, and clinical pregnancy rate were compared between the two groups. OBJECTIVE: We identified 22 active ingredients and 354 targets of action in Ziyin recipe, 791 targets of ovulatory infertility, and 96 common targets of action shared by Ziyin recipe and ovulatory infertility. According to Degree, the key targets included SRC, MAPK1, HSP90aa1, MAPK3, PTPN11, ESR1, Akt1, EGFR, NR3C1 and KNG1. Enrichment analysis of GO biological process showed that Ziyin recipe mainly focused on steroid hormonemediated signaling pathway, oxidation, reduction, and apoptosis. The most significantly enriched signaling pathways included PI3K-Akt signaling pathway, RAP1 signaling pathway, HIF-1 signaling pathway, estrogen signaling pathway, ovarian steroid production, and steroid hormone biosynthesis. The results of the clinical trial showed that Ziyin recipe significantly reduced the dose of GN, accelerated follicle development, increased E2 level of single dominant follicle on the trigger day, and increased the cycle ovulation rate and pregnancy rate. OBJECTIVE: The therapeutic effects of Ziyin recipe are probably mediated through different pathways to promote follicle development, thus improving ovulation rate and clinical pregnancy rate of infertile patients with ovulatory disorders.

First postpartum ovulation, metabolites and hormones in follicular fluid and blood in transition dairy cows supplemented with a Saccharomyces cerevisiae fermentation product.

We hypothesized that feeding a Saccharomyces cerevisiae fermentation product (SCFP) from -4 through +7 wk (calving = Day 0) facilitates early first postpartum ovulation and alters blood and follicular fluid concentrations of glucose, beta-hydroxybutyrate (BHB), free fatty acids (FFA), and steroid hormones favorable to subsequent fertility. Holstein cows were fed individually a SCFP product (n = 24) or served as controls (n = 23). Blood samples were collected at wk -4 and -2 from expected calving and at 1, 2, 5, and 7 wk postpartum to determine plasma concentrations of FFA and BHB. Early spontaneous ovulation (progesterone > 1 ng/mL or corpus luteum presence by postpartum median Day 33) or late ovulation was determined. Plasma FFA in weekly samples was not affected by SCFP supplementation, but FFA was greater (P < 0.01; week by ovulation status) in late compared with early ovulating cows during and after postpartum wk 2. Plasma BHB in weekly samples was greater (P = 0.03) in SCFP than control cows and tended (P = 0.06) to be greater in late than early ovulating cows. Cows were exposed to ovulation synchronization (GnRH, PGF2α, and GnRH on Days 33, 40, and 43 ± 3, respectively). Transvaginal dominant follicle aspiration was conducted at Day 50, 7 d after GnRH on Day 43. Metabolites (FFA, BHB, and glucose) and steroid hormones (progesterone, androstenedione, and estradiol) measured in follicular fluid and blood samples collected at aspiration revealed that androstenedione in serum was numerically less (P = 0.11) in SCFP-treated compared with control cows, whereas androstenedione in serum was less (P < 0.05) in late than early ovulating cows. Concentrations of BHB (r = 0.75) and glucose (r = 0.52) in follicular fluid were positively correlated (P < 0.01) with those in blood. Body weight at calving and Day 42 was less (P ≤ 0.05), and energy balance through Days 28 and 42 was more positive (P < 0.05) in early than late ovulating cows and in SCFP-supplemented compared with control cows (P < 0.05). Dry matter intake, daily milk yield, and yields of fat, protein, lactose, and total solids were less (P < 0.01) in early compared with late ovulating cows, whereas milk fat percentage was increased (P < 0.01) by SCFP supplementation. We conclude that elevated postpartum BHB and FFA in plasma, greater negative energy balance, and greater milk yield and components were associated with later postpartum ovulation, but metabolites and steroid hormones in blood and follicular fluid were unaffected by SCFP treatment or ovulation status except for androstenedione.

Lack of Efect of Short-term Lupin Grain Feeding on Ovulation Rate in Non-prolific Polish Mountain Ewes during the Breeding Season: Ultrasonographic and Endoscopic Assessment of Ovarian Activity.

The specific changes in antral follicle numbers and wave-like development have remained unrevealed in cyclic ewes fed high-protein, high-energy lupin grain for 6 days during the luteal phase of the estrous cycle (i.e., short-term nutritional flushing). This study was mainly conducted to determine ovarian effects of the 6-day lupin grain feeding in non-prolific Polish Mountain ewes, using transrectal ovarian ultrasonography and abdominal videoendoscopy. Estrus and ovulations were synchronized in 24 ewes with progestin-releasing intravaginal sponges for 12 days during the middle portion of the breeding season (September-October; 50.0458&deg;N, 19.8406&deg;E). Twenty-four ewes were assigned to three equal groups (n=8 each), including the Control group being fed the maintenance diet (i.e., hay-only), Treatment 1 receiving 500 g of lupin grain once a day, and Treatment 2 receiving 250 g of lupin grain twice a day, from days 9-14 of the synchronized estrous cycle (day 0=first ovulation of the interovulatory period studied). No differences were observed in the mean ovulation rate among the three groups of Polish Mountain ewes (P&gt;0.05). Ovarian antral follicles emerging in the penultimate wave of the estrous cycle in Treatment 2 ewes had a longer growth phase (p &lt;0.05) and attained a greater diameter (p &lt;0.05) before ovulation, in comparison to those in the other two groups. A final wave of the interovulatory interval emerged ~1 day earlier in Treatment 2 than in Treatment 1 ewes (p &lt;0.05). Nutritional supplementation with lupin grain increased the number of 3-mm follicles in Treatment 2 ewes (p &lt;0.05). The results of this study indicated that short-term nutritional flushing with lupin grain from mid- to late luteal phase did not consistently enhance ovulatory responses in non-prolific genotypes of ewes. Although the administration of lupins altered the timing of wave emergence, ovulatory follicle diameter, or duration of different stages of the follicular lifespan, it failed to increase the number of ovulatory follicles emerging in the penultimate and final waves of the estrous cycle in non-prolific Polish Mountain sheep.

Association between serum 25-hydroxy vitamin D level and menstrual cycle length and regularity: A cross-sectional observational study.

BACKGROUND: Research examining the role of vitamin D deficiency and the development of menstrual disorders in women is of widespread interest. Studies have been published showing that supplementation with high-dose vitamin D can lead to the restoration of the menstrual cycle. We lack adequate information regarding the effect of vitamin D levels on the physiology of menstruation and further on fertility in women of reproductive age due to the contradictory results reported by studies. OBJECTIVE: To study the association of 25-hydroxy vitamin D with menstrual cycle characteristics including long and short cycle length and cycle irregularity. MATERIALS AND METHODS: In this cross-sectional study, serum vitamin D levels of 166 women attending an outpatient department with menstrual irregularities after excluding all obvious causes of menstrual disorders (n = 83) between April-June 2019 were measured and were compared with women of similar profiles with complaints other than menstrual irregularities (n = 83). RESULTS: A decreased level of vitamin D was associated with a 13.3 times odds of an irregular cycle (OR (95% CI): 13.30 (5.79-30.60), p < 0.001). 25-hydroxy vitamin D was not associated with age or body mass index. We found a significant difference (p < 0.001) in mean vitamin D levels among the females with irregular cycles vs. regular cycles. CONCLUSION: Vitamin D plays a role in the physiology of reproduction specific to the menstrual cycle and ovulation. Long-term prospective studies assessing the exact cutoff value and the exact dose of supplementation required are needed.

Effects of Chinese herbal decoction on traditional Chinese medicine syndrome, menstrual cycle and sex hormone levels in amenorrhea patients with polycystic ovary syndrome / 中国基层医药

Objective:To investigate the effects of Chinese herbal decoction on traditional Chinese medicine syndrome, menstrual cycle and sex hormone levels in amenorrhea patients with polycystic ovary syndrome.Methods:A total of 124 amenorrhea patients with polycystic ovary syndrome who received treatment in The First People's Hospital of Lianyungang, China between December 2016 and December 2018 were included in this study. They were randomly assigned to receive either oral metformin hydrochloride and clomiphene citrate (control group, n = 62) or oral metformin hydrochloride and clomiphene citrate combined with Chinese herbal decoction (observation group, n = 62). Clinical effective rate and remission of traditional Chinese medicine syndrome were compared between control and observation groups. After treatment, recovery of menstrual cycle, ovulation rate, change of sex hormone level relative to before treatment, and the incidence of adverse reactions were determined in each group. Results:Total clinical effective rate in the observation group was significantly higher than that in the control group [96.77% (60/62) vs. 77.42% (48/62), χ2 = 12.817, P < 0.001]. Total effective rate in term of TCM syndrome in the control group was significantly higher than that in the observation group [83.87% (52/62) vs. 43.55% (27/62), χ2 = 21.800, P < 0.001]. After treatment, serum luteinizing hormone, free testosterone, luteinizing hormone / follicle stimulating hormone, estradiol, follicle stimulating hormone levels in the observation group were (4.28 ± 2.04) U/L, (1.93 ± 0.07) nmol/L, (3.17 ± 1.07), (240.32 ± 30.26) pmol/L, (3.17 ± 1.07) U/L, respectively, which were significantly lower than those in the control group [(6.45 ± 2.11) U/L, (3.27 ± 0.07) nmol/L, (5.73 ± 2.46), (300.32 ± 31.26) pmol/L, (5.12 ± 1.07) U/L), t = 5.822, 106.583, 7.514, 10.859, 10.147, all P < 0.001]. Ovulation rate in the observation group was significantly higher than that in the control group [80.65% (50/62) vs. 51.61% (32/62), χ2 = 11.666, P = 0.001]. Periodic ovulation rate in the observation group was significantly higher than that in the control group [91.94% (57/62) vs. 35.48% (22/62), χ2 = 14.528, P < 0.001]. In the observation group, the proportion of patients having edema, gastrointestinal reaction, vomiting, and diarrhea was 4.84% (3/62), 6.45% (4/62), 11.29% (7/62) and 11.29% (7/62), respectively, and they were 9.68% (6/62), 12.90% (8/62), 19.35% (12/62) and 25.80% (16/62)], respectively in the control group (all P < 0.05). Conclusion:Chinese herbal decoction can help regulate the hormone balance in amenorrhea patients with polycystic ovary syndrome, promote ovulation, eliminate amenorrhea, increase the curative effects on traditional Chinese medicine syndrome, and increase the rate of menstrual cycle recovery.