Serum AMH Levels Research Articles

The Comparison of Fixed and Flexible Progestin Primed Ovarian Stimulation on Mature Oocyte Yield in Women at Risk of Premature Ovarian Insufficiency

While gonadotrophin releasing hormone (GnRH) antagonists have been the standard of pituitary suppression during ovarian stimulation for ART, progestin primed ovarian stimulation (PPOS) has emerged as an alternative. Progestins can be started simultaneously with gonadotrophins (fixed PPOS) or later in the cycle depending on follicle growth (flexible PPOS). However, the flexible and fixed PPOS regimens have not been directly compared as of yet. This was a retrospective cohort study including women with diminished ovarian reserve who underwent oocyte cryopreservation. All women underwent ovarian stimulation with a fixed 300 IU daily dose of FSH. The primary outcome was the number of MII oocyte retrieved per cycle. Secondary outcome measures included the incidence of premature LH surge (>10ng/mL) and number of follicles larger than 14mm on the day of maturation trigger. During the screening period 2 out of 97 cycles were cancelled before oocyte retrieval, one in each group yielding an overall cancelation rate of 2%. Among women who had oocyte retrieval, 65 underwent flexible and 30 fixed PPOS. At baseline women on fixed and flexible PPOS had similar age (mean difference: -2.17 years, 95% CI: -4.46 to 0.11) and serum AMH levels (mean difference: 0.10 ng/mL, 95% CI: -0.24 to 0.47). Slight imbalances between the groups were rectified with propensity score matching using age and AMH levels. The incidence of premature LH surge (RR: 1.47, 95% CI: 0.51 – 5.27, p = 0.50), follicle count larger than 14mm on hCG day (RR: 1.14, 95% CI: 0.93 – 1.42, p = 0.22), number of MII oocytes retrieved (RR: 0.95, 95% CI: 0.79 – 1.15, p = 0.61) were similar between flexible and fixed PPOS. The rate of no oocyte retrieval was same between the groups (0.0% both) but no formal estimation was possible. Flexible and fixed PPOS regimens had no appreciable differences regarding MII oocyte yield and the incidence of premature LH surges. Cycles without oocyte retrieval were rare in both groups and ultrasonographic parameters of gonadotropin response were similar. Our study suggests the performances of either progestin regimen are comparable in this group of women.

Zuogui pills maintain the stemness of oogonial stem cells and alleviate cyclophosphamide-induced ovarian aging through Notch signaling pathway

BackgroundZuogui pills (ZGP), a classical prescription of traditional Chinese medicine, have been widely used in the treatment of ovarian aging. Previous studies have demonstrated its efficacy on protecting ovarian aging, and the mechanisms were mostly relevant to inhibiting the apoptosis of follicles and activating the primordial follicles. However, whether ZGP could stimulate the oogonial stem cells (OSCs) to refresh the follicle pool remains poorly understood. PurposeTo investigate the effects of ZGP on the stemness of OSCs in cyclophosphamide (Cy)-induced ovarian aging. Study design and methodsFemale Sprague-Dawley (SD) rats were randomly divided into 8 groups: control group, model group, ZGP groups (low / high dose groups), estradiol valerate (EV) groups (low / high dose groups), DAPT group and DAPT+ZGP-L group. After modeling with Cy, the ZGP groups and EV groups were treated with ZGP and EV for 8 weeks respectively. Meanwhile, the DAPT groups were treated with DAPT twice a week. Additionally, OSCs were also isolated after modeling, and then treated with drug serum containing ZGP or EV. Ovarian volume and the ratio of weight of total ovaries to the body weight were measured. The serum hormones were measured by ELISA. Quantities and location of OSCs in ovaries were detected by flow cytometry and immunofluorescence. Cell viability was measured by CCK8. And OSCs were identified by immunofluorescence. Biomarkers of germ cells, stem cells and associated to differentiation and meiosis were detected by qPCR and western blot. Proteins in Notch signaling pathway were detected by western blot and immunofluorescence. ResultsAfter treating with ZGP, ovarian volume and the ratio of weight of total ovaries to the body weight increased. ZGP could increase serum AMH and E2 level and decrease serum FSH level. Quantities and cell viability of OSCs increased after ZGP treatment in vivo and in vitro. In addition, treatment with ZGP could increase not only the expression of MVH, Oct4 and DAZL, but also the expression of ZP1 and ZP2. Furthermore, ZGP could up-regulate the expression of Notch intracellular domain (NICD), HES1 and HES5. After blocking the Notch signaling pathway, ZGP could increase not only the expression of NICD, HES1 and HES5, but also the expression of MVH, Oct4, DAZL, ZP1 and ZP3. ConclusionIn conclusion, the mechanism of ZGP on treating ovarian aging may be relevant to maintain the stemness of OSCs by up-regulating Notch signaling pathway, which added the mechanism of ZGP on the perspective of OSCs at first time.

Laparoscopic ovarian drilling before IVF/ICSI in polycystic ovarian syndrome patients with high antimullarian hormone

ObjectiveTo evaluate the effect of performing laparoscopic ovarian drilling [LOD] before proceeding to in vitro fertilization [IVF] or intracytoplasmic sperm injection [ICSI] on the cycle outcomes in polycystic ovarian syndrome [PCOS] patients with high antimullerian hormone [AMH] levels. Study designA retrospective case-control study. Patients and methodsThe study was conducted in the Fertility Care Unit in our University Hospital. The participants of the study were PCOS women with high serum AMH levels [≥ 7 ng/ml] who were subjected to IVF/ICSI. Women who underwent LOD in the preceding 3 months before IVF/ICSI [study group; n = 76] were compared with a control group of women who did not undergo LOD [control group; n = 71]. ResultsTotal gonadotropin consumption and the number of stimulation days were significantly lower with LOD [P = 0.026 and 0.010, respectively]. Oocyte maturity rate was significantly higher with LOD [P = 0.028]. The incidence of severe OHSS was significantly lower with LOD [P = 0.026]. No significant difference between both groups in the pregnancy and implantation rates. ConclusionThe LOD before IVF/ICSI in PCOS patients with high AMH [≥ 7 ng/ml] has the benefits of reduction in gonadotropins consumption and stimulation days with an increase in the percentage of collected mature oocytes and decrease in the incidence of severe OHSS; however, it seems not to improve the pregnancy and implantation rates.

Comparative advantage of anti-Mullerian hormone over other ovarian reserve metric (basal hormonal test) in prediction of fertility in women with varying menstrual cycle

Objective: Anti-Mullerian hormone has been indicated as a novel biomarker for ovarian reserve assessment. This study aimed to determine the comparative advantage of serum levels of AMH, FSH, LH, E2, and LH/FSH ratio among women with varying menstrual cycles and duration of menstruation. Material and Method: A total of 90 subjects, which consisted of sixty subjects and thirty healthy subjects as control, were recruited. Blood samples were collected on day 3 of the menstrual cycle and evaluated for ovarian markers using the ELISA technique. All data were analyzed using SPSS version 23.0. Results: AMH and LH/FSH levels were lower in women with varying menstrual cycles than in the control group. FSH, LH, and E2 levels were significantly higher in women with varying menstrual cycles and flow duration than in the control group(P<0.05). AMH was negatively correlated with age (r= -0.72), BMI( r= -0.4), FSH(r= - 0.3), LH(r = -0.2) (p<0.05) and E2(r= -0.2, p>0.05). Also age was positively correlated with FSH(r=0.5,p<0.05), E2(r=0.3, p<0.05) and BMI(r=0.4, p<0.05). The level of AMH was not significant with cycle length and days of flow(p>0.05). This implies that AMH can be measured independently of the cycle phase. This show that AMH was lower in women with varying menstrual cycle with an increase in gonadotrophin and E2. The strong negative relationship between age and AMH implies that age is determining factor of ovarian reserve. Conclusion: AMH combined with age and FSH may improve ovarian reserve evaluation, making AMH a better marker

Antimullerian Hormone Levels and Association with Abortion and Preterm Delivery in Patients with Polycystic Ovary Syndrome Who Conceived with Assisted Reproductive Techniques.

Pregnant PCOS patients were found to suffer from many adverse outcomes of pregnancy. Prediction of the fate of pregnancy and labor in PCOS patients was highly needed. There were recently discovered roles of serum AMH in those patients who were seeking for pregnancy and who were conceiving with assisted reproductive techniques. To analyze the predictive roles of measuring serum levels of AMH in patients with PCOS who became pregnant spontaneously or used assisted reproductive techniques regarding abortion and preterm delivery. A total of 100 females with PCOS were included in the study and they underwent assisted reproductive techniques were included in the study and underwent measurement of AMH levels regularly. We found that a total of 70 patients had a term delivery, and 30 patients had a preterm delivery. We found no statistically significant differences between both groups regarding their age or body mass index (BMI). We showed that serum AMH levels were higher in the group of PCOS patients who had preterm delivery than in the group of patients with term delivery (p < .0.001). High serum AMH levels were found to be associated with higher risks of occurrence of preterm labor in patients with PCOS who underwent assisted reproductive techniques. Our results gave a clue to clinicians for better management of the pregnancy process in these patients.

Perinatal Outcomes In Monozygotic Pregnancies Resulting From Assisted Reproductive Technology Procedures: A Single-Center 6-Year Experience Based On A Large Cohort Of Pregnancies.

Monozygotic twin (MZT) pregnancies increase the risk of maternal and infant mortality and include many complications. The present study describes our assisted reproductive technology (ART) procedures from the viewpoint of perinatal outcomes in MZT pregnancies. In this retrospective clinical cross-sectional study, 1159 in vitro fertilization (IVF) cycles performed between October 2014 and December 2019 were reviewed and perinatal outcomes and general clinical conditions analyzed. Sixteen MZT pregnancies were observed, resulting in an incidence of 1.38%. The MZT pregnancy incidence for patients aged ≤35 and >35 years were 0.2% and 1.1%, respectively. Eight MZT pregnancies resulted in live births, while five ended in miscarriage. A significant positive correlation was found between the number of attempts and the age of female (r:0.674; p=0.004) and male (r:0.657; p=0.006) partners. Cumulus-Oocytes Complexes (COC) (r:0.635; p=0.008), Metaphase II Oocyte (MIIO) (r:0.627; p=0.009), Pronucleus Oocyte (PO) (r:0.585; p=0.017) were correlated with serum AMH levels. The number of MZT was positively correlated with male partner age (r:0.527; p=0.036) and negatively correlated with embryo transfer day (ETd) (r:-0.548; p=0.028). The incidence of MZT pregnancies observed in this study was similar to the incidence reported in the literature, although risk was more pronounced among women aged >35 years. Due to potential risks for mothers and fetuses, MZT pregnancies may become a problem as the number of individuals seeking IVF continues to increase.

Melatonin protects against ovarian damage by inhibiting autophagy in granulosa cells in rats

ObjectivesThis study sought to further verify the protective mechanism of Melatonin (MT) against ovarian damage through animal model experiments and to lay a theoretical and experimental foundation for exploring new approaches for ovarian damage treatment. MethodThe wet weight and ovarian index of rat ovaries were weighted, and the morphology of ovarian tissues and the number of follicles in the pathological sections of collected ovarian tissues were recorded. And the serum sex hormone levels, the key proteins of the autophagy pathway (PI3K, AKT, mTOR, LC3II, LC3I, and Agt5) in rat ovarian tissues, as well as the viability and mortality of ovarian granulosa cells in each group were measured by ELISA, western blotting, CCK8 kit and LDH kit, respectively. ResultsThe results showed that MT increased ovarian weight and improved the ovarian index in ovarian damage rats. Also, MT could improve autophagy-induced ovarian tissue injury, increase the number of primordial follicles, primary follicles, and sinus follicles, and decrease the number of atretic follicles. Furthermore, MT upregulated serum AMH, INH-B, and E2 levels downregulated serum FSH and LH levels in ovarian damage rats and activated the PI3K/AKT/mTOR signaling pathway. Besides, MT inhibited autophagic apoptosis of ovarian granulosa cells and repressed the expression of key proteins in the autophagic pathway and reduced the expression levels of Agt5 and LC3II/I. ConclusionsMT inhibits granulosa cell autophagy by activating the PI3K/Akt/mTOR signaling pathway, thereby exerting a protective effect against ovarian damage.

Exploration of the Danggui Buxue Decoction Mechanism Regulating the Balance of ESR and AR in the TP53-AKT Signaling Pathway in the Prevention and Treatment of POF

Objective The purpose of this study was to explore the molecular mechanism of Danggui Buxue Decoction (DBD) intervening premature ovarian failure (POF). Methods The active compounds-targets network, active compounds-POF-targets network, and protein-protein interaction (PPI) network were constructed by a network pharmacology approach: Gene Ontology (GO) function and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway analysis by DAVID 6.8 database. The molecular docking method was used to verify the interaction between core components of DBD and targets. Then, High-Performance Liquid Chromatography (HPLC) analysis was used to determine whether the DBD contained two key components including quercetin and kaempferol. Finally, the estrous cycle, organ index, ELISA, and western blot were used to verify that mechanism of DBD improved POF induced by cyclophosphamide (CTX) in rats. Results Based on the network database including TCMSP, Swiss Target Prediction, DisGeNET, DrugBank, OMIM, and Malacard, we built the active compounds-targets network and active compounds-POF-targets network. We found that 2 core compounds (quercetin and kaempferol) and 5 critical targets (TP53, IL6, ESR1, AKT1, and AR) play an important role in the treatment of POF with DBD. The GO and KEGG enrichment analysis showed that the common targets involved a variety of signaling pathways, including the reactive oxygen species metabolic process, release of Cytochrome C from mitochondria and apoptotic signaling pathway, p53 signaling pathway, the PI3K-Akt signaling pathway, and the estrogen signaling pathway. The molecular docking showed that quercetin, kaempferol, and 5 critical targets had good results regarding the binding energy. Chromatography showed that DBD contained quercetin and kaempferol compounds, which was consistent with the database prediction results. Based on the above results, we found that the process of DBD interfering POF is closely related to the balance of ESR and AR in TP53-AKT signaling pathway and verified animal experiments. In animal experiments, we have shown that DBD and its active compounds can effectively improve estrus cycle of POF rats, inhibit serum levels of FSH and LH, protein expression levels of Cytochrome C, BAX, p53, and IL6, and promote ovary index, uterine index, serum levels of E2 and AMH, and protein expression levels of AKT1, ESR1, AR, and BCL2. Conclusions DBD and its active components could treat POF by regulating the balance of ESR and AR in TP53-AKT signaling pathway.

Effect of GnRH agonist combined with menstrual blood-derived stem cell transplantation on ovarian function in mice

To investigate the effect of combined treatment with gonadotropin-releasing hormone (GnRH) agonist and menstrual blood-derived stem cell (MenSC) transplantation on ovarian function in mice. We tested the effect of GnRH agonist and MenSC transplantation, either alone or in combination, in a mouse model of premature ovarian failure (POF) induced by daily intraperitoneal injection of 120 mg/kg cyclophosphamide (CTX) for 15 days. Venirelin was administered at the daily dose of 0.1 mg via intravenous infusion, starting at 14 days before modeling; MenSC suspension (2× 106 cells) was injected into the ovarium 1 day after modeling. HE staining was used to observe ovarian pathologies of the mice. ELISA was used to detect serum levels of LH, FSH, AMH and E2. The expression levels of ki-67, Bcl-2, BAX, caspase 9, caspase 3, their cleavages, P-PI3K, PI3K, P-Akt and Akt proteins in ovarian granulosa cells were determined with Western blotting. The mouse models of POF showed obvious ovarian fibrosis with increased atresia follicles and had significantly elevated serum levels of LH and FSH (P < 0.01), decreased serum levels of AMH and E2 (P < 0.01), significantly increased ovarian expression levels of caspase 9, caspase 3, and their cleavages (P < 0.01), and decreased expression levels of Ki-67, Bcl-2, P-PI3K and P-Akt (P < 0.01). The combined treatment with GnRH agonist and MenSC transplantation obviously inhibited the production of ovarian atresia follicles, lowered serum levels of LH and FSH (P < 0.01), and increased the levels of AMH and E2 (P < 0.05 or 0.01) in the mouse models. The combined treatment also inhibited the expressions of BAX and cleaved caspase 9 and caspase 3 proteins (P < 0.05 or 0.01) and enhanced the expressions of Ki-67, Bcl-2, P-PI3K, and p-Akt proteins (P < 0.05 or 0.01) in ovarian granulococcus cells. In mouse models of POF, GnRH agonist combined with MenSC transplantation can reduce the generation of ovarian atresia and improve ovarian reserve by up-regulating PI3K-Akt signaling pathway to achieve ovarian repair and protection.

Effects of quercetin on ovarian function and regulation of the ovarian PI3K/Akt/FoxO3a signalling pathway and oxidative stress in a rat model of cyclophosphamide-induced premature ovarian failure.

To investigate the ability of quercetin to improve ovarian function and inhibit ovarian oxidative stress through the PI3K/Akt/FoxO3a signalling pathway in a rat model of premature ovarian failure (POF), we constructed a POF rat model with cyclophosphamide (CTX) and treated it with quercetin. Haematoxylin and eosin staining (H&E staining) was used to observe the morphological changes of the ovaries. The serum levels of AMH, E2, FSH, SOD, GSH-Px and MDA were determined by enzyme-linked immunosorbent assays. The expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), forkhead box O3a (FoxO3a) and their phosphorylated forms AMH, FSH and their receptors in the ovary were detected by western blots. The mRNA expression of PI3K, Akt, FOXO3a, AMH, FSH and their receptors was detected by qRT-PCR. Our results showed that quercetin could significantly increase the expression of AMH, E2, SOD and GSH-Px, upregulate the protein expression of AMH, FSH and its receptor and decrease the expression ratio of phosphorylated PI3K, Akt, FOXO3a and the unphosphorylated forms. Moreover, quercetin inhibited the mRNA expression of PI3K, Akt and FOXO3a. These results suggest that quercetin can restore ovarian function and inhibit oxidative stress by regulating the PI3K/Akt/FoxO3a signalling pathway.

Anti-Müllerian hormone was independently associated with central obesity but not with general obesity in women with PCOS.

ObjectiveAnti-Müllerian hormone (AMH) is recognized as the most important biomarker for ovarian reserve. In this cross-sectional study, we aimed to explore the potential association of AMH with central obesity or general obesity in women with polycystic ovary syndrome (PCOS).MethodsIn this cross-sectional study, 179 patients with PCOS were enrolled and underwent anthropometric measurements (BMI and waist circumference (WC)) and serum AMH level detection. Pearson’s correlation and multivariable logistic regression analyses were performed to determine the associations of AMH with central obesity and general obesity.ResultsSubjects with increasing BMI showed significantly lower values of AMH (median (interquartile range (IQR)) 8.95 (6.03–13.60) ng/mL in normal weight group, 6.57 (4.18–8.77) ng/mL in overweight group, and 6.03 (4.34–9.44) ng/mL in obesity group, P = 0.001), but higher levels of systolic blood pressure, fasting insulin, total cholesterol, triglycerides, LDL-c, obesity indices (WC, hip circumferences, waist-to-hip ratio, waist-to-height ratio (WHtR), and Chinese visceral adiposity index (CVAI)). Compared with the group of PCOS women without central obesity, the group with central obesity had significantly lower value of AMH (median (IQR) 8.56 (5.29–12.96) ng/mL vs 6.22 (4.33–8.82) ng/mL; P = 0.003). Pearson’s correlation analysis showed that AMH was significantly and negatively correlated with BMI (r = −0.280; P < 0.001), WC (r = −0.263; P < 0.001), WHtR (r = −0.273; P < 0.001), and CVAI (r = −0.211; P = 0.006). Multivariate logistic regression analysis with adjustment for potential confounding factors showed that AMH was independently and negatively associated with central obesity but was not significantly associated with general obesity.ConclusionsAMH was independently and negatively associated with central obesity. Closely monitoring the WC and AMH should be addressed in terms of assessing ovarian reserve in women with PCOS.

Pregnancy outcome of freeze thaw cycles of polycystic ovary syndrome patients regarding the anti-Müllerian hormone percentile

Patients with polycystic ovary syndrome display increased levels of anti-Müllerian hormone. Frozen-thawed embryo transfer (FET) prevents ovarian hyperstimulation and results in better pregnancy outcome in PCOS patients. Therefore, we aimed to evaluate the effect of serum AMH levels on the pregnancy outcome of FET cycles in PCOS patients. 110 infertile women with PCOS who were recommended for embryo cryopreservation followed by FET. The patients’ AMH levels were evaluated, and the age-related AMH percentiles were determined. The patients were then grouped according to AMH percentiles, namely, 75th–90th percentile (Group 1) and higher than the 90th percentile (Group 2). A total of 110 PCOS patients who conceived in Frozen Embryo Transfer (FET)-In-vitro Fertilisation (IVF) cycles were included in this study. The preterm delivery rates in FET cycles were higher in the group of PCOS patients with AMH levels greater than the 90th percentile than in patients in the 75th–90th percentile group (50% vs 28.8%, p = .024). In conclusion, PCOS patients with AMH levels higher than the 90th percentile had substantially higher preterm delivery rates than those with AMH levels at the 75th–90th percentile, suggesting the need for closer follow-up. Further studies are needed to elucidate the underlying mechanisms behind this correlation. IMPACT STATEMENT What is already known on this subject? The association of AMH levels with the risk of adverse pregnancy outcomes has been previously investigated. In women with PCOS, substantially elevated AMH levels were significantly associated with preterm birth. What do the results of this study add? Results showed that the PCOS patients with higher AMH levels and underwent assisted reproductive treatment demonstrated an increased risk of preterm labour than the PCOS patients with lower AMH levels. What are the implications of these findings for clinical practice and/or further research? In women with PCOS, substantially elevated AMH levels were significantly related to preterm birth, suggesting the need for closer follow-up in this population and the need for further studies to elucidate the underlying mechanisms behind this correlation.

Effect of acupuncture for diminished ovarian reserve: study protocol for a randomized controlled trial

BackgroundDiminished ovarian reserve (DOR) is a condition in which the ovary loses its normal reproductive potential, compromising fertility. Although the prevalence and incidence of DOR is increasing, there are currently no effective treatments for this condition. Acupuncture has been reported as an alternative therapy for female infertility. The purpose of this study is to investigate the effect of acupuncture for women with DOR.Methods/designIn this randomized controlled trial, a total of 120 women with DOR will be randomly assigned to receive either acupuncture or sham acupuncture for 12 weeks. The primary outcome will be determined by the mean change from baseline in the antral follicle count (AFC) at week 12. Secondary outcomes include serum levels of FSH, LH, E2, and AMH, the length of menstrual cycle, and the score of Self-Rating Anxiety Scale (SAS).DiscussionThis study is expected to investigate the effectiveness of acupuncture versus sham acupuncture in improving ovarian reserve for women with DOR.Trial registrationAcupuncture-Moxibustion Clinical Trial Registry ChiCTR1800014988. Registered on 6 February 2018

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.

Effects and mechanisms of mUCMSCs on ovarian structure and function in naturally ageing C57 mice

BackgroundThe ovaries are the core reproductive organs in women and are critical for maintaining normal reproductive function and endocrine system stability. An ageing C57 mouse model was used to evaluate the efficacy and mechanism of mouse umbilical cord mesenchymal stem cells (mUCMSCs) and to explore the mechanism by which mUCMSCs promote the antioxidant repair of mouse granulosa cells (mGCs).ResultsEighteen-month-old C57 mice were randomly divided into a model group and a treatment group. At the same time, 2-month-old C57 mice were established as a young group (15 mice per group). The mice in the treatment group were injected via the tail vein with GFP-labelled mUCMSCs. The ovarian volume in ageing C57 mice was decreased, and there were no follicles at any stage. After mUCMSC transplantation, the mouse ovaries increased in size, follicles at various stages were observed in the cortex, and the antral follicle counts increased. The serum E2, AMH, and INH-B levels of mice in the treatment group were significantly higher than those of mice in the model control group (P < 0.05). mUCMSCs downregulated the expression of the autophagy-related gene LC3b and the apoptosis-related genes Bax and Caspase-3, upregulated the expression of SOD2 and the peroxidase gene PRDX IV, and reduced apoptosis rates and reactive oxygen species (ROS) levels in granulosa cells.ConclusionsmUCMSCs play roles in promoting the repair of ageing ovaries by regulating immunity, anti-inflammatory responses and the PI3K-Akt signalling pathway.

IMPACT OF SURGERY ON OVARIAN RESERVE IN PATIENTS WITH ENDOMETRIOMA AMONG INFERTILE WOMEN

To find out the correlation between pre-operative serum AMH and FSH levels and 3 months post-operative levels. The study was conducted in the Reproductive Endocrinology and Infertility Unit of Dhaka Medical College and Hospital (DMCH), Dhaka, Bangladesh, during July 2018 to December 2019. All admitted patients diagnosed with ovarian endometriomas (>4 cm) and of 20-35 years of age were included. Laparoscopy was the mainstay of surgery but in a few cases laparoscopy followed by laparotomy was done. Cystectomy (complete excision of the capsule) was attempted and adhesiolysis was also performed. Electrocoagulation was needed in few cases for haemostasis, which was performed by monopolar or bipolar diathermy. Carbon dioxide laser vaporization was not available, and cyst drainage was not done. In open cases, reconstruction of the ovary was performed. This study was a descriptive cross-sectional study. Initially for all patients, Serum AMH levels were measured preoperatively. Three months post surgery, Serum AMH was measured again, and data analysis and comparison of these values with ovarian reserve was done. Change in Serum FSH levels was also compared. Student’s t-test, paired and unpaired, were performed for different data. 69 patients were studied. 52.17% had tumours 4-6 cm, >6 cm in 47.83%. Tumour was bilateral in 36.23%. Laparoscopic cystectomy by monopolar cauterization was the 55.0% patients, laparotomy required in 10.14% patients. Average pre-operative serum AMH level in <29 yrs patients was 3.06±1.35 ng/ml, 2.38±1.54 postoperatively (p<0.001). For 30-35 yrs, they were 2.30±1.16 ng/ml and 1.39±1.01 ng/ml respectively (p<0.001). No significant difference in AMH was found in unilateral and bilateral cases (p=0.842), significant differences were found in pre- and postcystectomy AMH levels in these groups (p<0.001 for both). Pre and post-surgery FSH differences were very significant, p<0.001. When analysing the effect of different types of surgery we found that coagulation and excision both have negative effect on serum AMH levels, after 3 months (p<0.001). But there is no significant difference postsurgery between these two groups (p=0.0915). Surgery has a negative impact on ovarian reserve in patients with endometrioma, as evident by analysing different metrics and utilising student’s t-test.

LONGITUDINAL EXAMINATION OF THE ASSOCIATION BETWEEN HORMONAL CONTRACEPTIVE USE AND ANTI-MÜLLERIAN HORMONE (AMH) LEVELS IN REPRODUCTIVE-AGED AFRICAN-AMERICAN WOMEN

To determine the impact of hormonal contraceptive (HC) use on trends in AMH levels in a longitudinal cohort of reproductive-aged African American women (AAW). This study used an established cohort of AAW from Detroit, MI aged 25 to 35 years, who were recruited as a part of the Study of the Environment, Lifestyle and Fibroids. Anthropometric measurements, self-reported health history, and serum AMH levels (picoAMH assay, Ansh Labs, Webster, TX) were collected from participants at 4 visits over 5 years. All types of HC were included. Percent changes in AMH levels were calculated from baseline to final visit and compared across groups with different patterns of HC use using linear regression models, adjusting for age, BMI, history of abnormal menstrual bleeding, thyroid disorder, and a history of infertility. The effect of starting, continuing, or stopping HC on rate of change in AMH for all follow up visits was also assessed using linear mixed models adjusted for age. A total of 1531 women were included with a mean age of 29.2 ± 3.4 years and a median AMH of 4.07 (IQR 2.29 - 6.70) ng/mL at baseline. At the final follow-up visit, mean age was 34.1 ± 3.4 years and median AMH was 2.97 (IQR 1.52 - 5.15) ng/mL, with a 19.8% average decline in AMH. Women who started and continued HC during the study period had significantly greater % decrease in AMH levels (-23.1%, 95% CI -40.3%, -5.9%) compared with women who never used HC. When compared with women who never used HC, the % change in AMH levels among women who started and stopped HC during the study was -14.4% (95% CI -29.3%, 0.5%), among women who only used HC prior to the study was -13.5%, (95% CI -26.9%, -0.1%) , and among long-term users of HC (on HC before and throughout study period) was -8.7% (95% CI -24.6%, 7.2%). We next sought to determine whether the rate of decline in AMH was impacted by the recency of HC start or discontinuation. Women who started HC between study visits had a faster rate of decline in AMH levels compared with women not on HC at their previous or current visit, (-11.9%; 95% CI -15.5%, -8.0%), while women who used HC at a previous visit but discontinued prior to the most recent visit had a slower rate of decline in AMH (10.7% 95% CI 6.7%, 15.0%). There was no difference in rate of decline among women who continued HC between visits (2.3%, 95% CI -0.6%, 5.7%) compared to those who were not on HC between visits. In this study, the % decline in AMH levels with age was greatest among those who started and continued HC during the study period. This finding, along with the non-significant difference in % decrease with age in AMH levels among long-term HC users, suggests that the impact of HC use on AMH may be most significant after initiation with recovery of AMH following stoppage of HC suggested by the smaller decline with age in this subset of women. Thus, the impact of HC on AMH levels appears reversible. In addition, being on HC neither increases nor protects against the age related rate of decline.

GROUP BASED TRAJECTORY MODELING (GBTM) OF ANTI-MULLERIAN HORMONE (AMH) IN A COHORT OF REPRODUCTIVE AGE AFRICAN AMERICAN WOMEN (AAW)

AMH has been used as a marker of ovarian reserve. Limited data are available to observe longitudinal changes in AMH over time. Our objective was to evaluate group based AMH trajectory models in a group of reproductive age AAW over the course of 5 years. This longitudinal study used an established cohort of AAW from Detroit, MI aged 23-35 years, who were recruited as a part of the Study of the Environment, Lifestyle and Fibroids. Anthropometric measurements, self reported medical history, and serum AMH levels (picoAMH assay, Ansh Labs, Webster, TX) were collected from participants at 4 visits over 5 years. GBTM was used to establish clusters of women with similar AMH trajectories using age as the time variable. The number of groups and form of temporal trend (linear, quadratic, etc.) for each group trajectory was determined using Bayes Information Criteria. Time varying BMI and hormonal contraception (HC) use were tested to determine their effect on group trajectories. AMH is not normally distributed so analyses were carried out on the logarithmic scale and back transformed to report predicted AMH. 1551 women with AMH for at least 2 visits were included with mean baseline age of 29.2 ± 3.4 years and median AMH of 4.08ng/mL (IQR 2.28 - 6.70). At visit 4, mean age was 34.2 ± 3.4 years and median AMH was 3.00ng/mL (1.52 - 5.15). GBTM model results indicate 5 different groups of AMH trajectories in this population. Group 1 (0.9% of participants) has a linear decline in log AMH with predicted AMH of 0.03ng/mL at age 23 and 0.001ng/mL at age 35 for a normal weight woman not using HC. Group 2 (6.7% of participants) has a cubic trajectory with an AMH value of 1.80ng/mL at age 23 and 0.31ng/mL at age 35. Groups 3, 4, and 5 (comprising 27.8%, 45.4%, and 19.3% of participants respectively) all had a quadratic trajectory for log AMH, although with different slopes. In group 3, at age 23 the predicted AMH is 2.87ng/mL while at age 35 is 1.37ng/mL; for group 4 the predicted AMH at age 23 is 6.13ng/mL while at age 35 is 3.61ng/mL, and for group 5 the predicted AMH at age 23 is 13.8ng/mL and at age 35 is 8.7ng/mL. Time varying HC use and overweight or obese status is associated with AMH trajectories in some groups. In groups 3, 4, and 5, HC use is associated with 15.4% (p<0.001), 15.5% (p<0.001), and 23.9% (p<0.001) lower AMH compared to not using HC, respectively. Being obese is associated with 10.2% (p=0.016) and 12.1% (p=0.029) lower AMH compared to normal weight in groups 4 and 5, respectively. Group 1, with very low AMH at all 4 visits, is associated with 846.6% (p<0.001) higher AMH in HC users, 258.8% (p<0.001) higher AMH in overweight women, and 611.4% (p<0.001) higher AMH in obese women. We identified five groups of AMH trajectories over time. HC and overweight or obese status were found to influence AMH trajectories in some, but not all, groups. Further study is needed to understand other factors that may impact AMH trajectories or risk factors for group membership.

DOES DUAL TRIGGER WITH COMBINATION OF GONADOTROPIN-RELEASING HORMONE AGONIST AND LOW DOSE HUMAN CHORIONIC GONADOTROPIN IMPROVES CLINICAL OUTCOMES IN NORMAL RESPONDERS IN GnRH ANTAGONIST IVF-ICSI CYCLES

Clinical outcomes of dual trigger containing gonadotropin-releasing hormone agonist (GnRHa) and low dose human chorionic gonadotropin (hCG) compared with the standard dose of human chorionic gonadotropin (hCG) trigger for final oocyte maturation in normal responder patients in GnRH Antagonist IVF-ICSI Cycles. A single centre prospective randomized comparative study conducted at a tertiary care infertility centre , Akanksha IVF centre, Janakpuri, New Delhi from 1st March 2020 to 28th February 2021. 80 normal responder patients defined as women in age group 21 to 37 year, with serum AMH level >1.2 ng/ml to < 4ng/ml , with AFC (3- 8) per ovary who were to undergo controlled ovarian hyperstimulation (COH) for IVF-ICSI cycles, were randomized by a computer based program into 2 groups of 40 each. All the patients underwent ovarian stimulation with a starting dose of recombinant FSH with dosage 150-225 IU on D2 and the dosage were adjusted according to follicular monitoring After at least one follicle reached a size of 14mm , antagonist Inj Cetrorelix 0.25 mg started subcutaneously daily. When > 2 follicles reached the size of 18 mm final oocyte maturation was triggered by either dual trigger of Inj Leuprolide acetate 2 mg and low dose 2000 IU highly purified Inj. hCG subcutaneously in Group 1 or by standard trigger dose 10000 IU of highly purified inj hCG in Group2. Oocyte retrievals were performed 35 to 36 hours after the trigger. IVF-ICSI was performed in all cases. Day 3 fresh embryo transfers were performed for all cases (2 x 8cell embryos).The luteal phase support was with vaginal supplementation of 800 mg micronized progesterone. Serum beta hCG was performed after14 days of embryo transfer . Primary outcome measured was clinical pregnancy rate and secondary outcomes measured were: implantation rate, miscarriage rate, number of MII oocytes picked up, number of embryos formed, and risk of OHSS. In dual trigger , group 1 the clinical pregnancy rate (52.50% vs 47.50% ) and implantation rate (29.67% vs 26.08% ) were higher than in Group 2 , and but the difference was not statistically significant. Miscarriage rate was lesser in group 1 (14.28%) than in group 2 (15.78%) and it was also not statistically significant . There were higher number of MII oocytes picked up and higher number of embryos formed in group1/ dual trigger group (10.63+5.46 and 8.2+3.4 respectively ) than in group 2 (8.10+5.74 and 6.8+3.6 respectively) and this difference was statistically significant . No OHSS was reported in group 1/ dual trigger group. Though there was increased clinical pregnancy rate, implantation rate and a lesser miscarriage rate in dual trigger group1 for but it was statistically not significant .There was significant increase in MII oocytes and number of embryos formed (p value .0064 and .0 156 respectively) in the dual trigger group. There was also no risk of OHSS in dual trigger group.

CAN A HIGH AMH OVERCOME ADVANCING REPRODUCTIVE AGE? THE ASSOCIATION BETWEEN AMH AND CUMULATIVE LIVE BIRTH AMONG WOMEN OVER AGE 40

Serum AMH levels are routinely used to predict an individual’s response to controlled ovarian stimulation. The association between AMH and live birth is controversial, however, particularly in older women undergoing treatment with in vitro fertilization (IVF). Previous studies were limited by the exclusion of cycles with preimplantation genetic testing (PGT-A) and frozen embryo transfer (1, 2). Among older women, those with higher AMH may be more likely to achieve blastocysts for biopsy or cryopreservation.