Elevated Serum Progesterone Levels Research Articles

Alterations in the Hypothalamic-Pituitary-Adrenal Axis as a Response to Experimental Autoimmune Encephalomyelitis in Dark Agouti Rats of Both Sexes.

Multiple sclerosis (MS) is a chronic inflammatory disease that affects the central nervous system, usually diagnosed during the reproductive period. Both MS and its commonly used animal model, experimental autoimmune encephalomyelitis (EAE), exhibit sex-specific features regarding disease progression and disturbances in the neuroendocrine and endocrine systems. This study investigates the hypothalamic-pituitary-adrenal (HPA) axis response of male and female Dark Agouti rats during EAE. At the onset of EAE, Crh expression in the hypothalamus of both sexes is decreased, while males show reduced plasma adrenocorticotropic hormone levels. Adrenal gland activity is increased during EAE in both males and females, as evidenced by enlarged adrenal glands and increased StAR gene and protein expression. However, only male rats show increased serum and adrenal corticosterone levels, and an increased volume of the adrenal cortex. Adrenal 3β-HSD protein and progesterone levels are elevated in males only. Serum progesterone levels of male rats are also increased, although testicular progesterone levels are decreased during the disease, implying that the adrenal gland is the source of elevated serum progesterone levels in males. Our results demonstrate a sex difference in the response of the HPA axis at the adrenal level, with male rats showing a more pronounced induction during EAE.

P-648 How often does elevated progesterone occur? A systematic review

Abstract Study question What is the incidence of elevated serum progesterone level during the follicular phase in fresh and frozen ART cycles? Summary answer The incidence of elevated progesterone levels varies widely during the follicular phase in both fresh and frozen ART cycles. What is known already Progesterone monitoring is often performed for the optimization of ART cycle. Elevated progesterone (EP) noted during ovarian stimulation can lead to cycle cancellation and the implementation of a freeze all strategy as EP is associated with lower pregnancy rates. The introduction of GnRH analogues did not seem to prevent EP. In frozen embryo transfer (FET) cycles, EP can be present even without gonadotropin commencement. Various thresholds have been used to define EP and the incidence of EP reported in the literature varies. Study design, size, duration A systematic search of the following electronic databases: EMBASE, MEDLINE, CINAHL and PubMed identified non-randomised comparative cohort studies between the year 2000 to 2022. The following keywords progesterone, assisted reproductive technique and pregnancy outcomes and their respective variants were used. Study selection was performed following initial screen of the titles and abstracts. The full manuscripts were examined for compliance with the inclusion criteria and studies eligible for inclusion in the review were selected. Participants/materials, setting, methods PICOS study protocol was used. We included studies reporting per woman data of women undergoing fresh IVF/ICSI cycles (with controlled ovarian stimulation) and FET cycles (natural or medicated), with extractable data on pregnancy outcomes and using serum progesterone monitoring. We excluded interventional studies and studies involving oocyte or gamete donation. Comparisons were made between the population with EP versus non-elevated progesterone in fresh and FET cycles. Main results and the role of chance We included 52 studies (N = 53,517 women) for the systematic review. In fresh COS cycles, the incidence of EP at baseline varied from 13.3% to 30% (30% in P > 0.65ng/ml; 13.3% - 23.2% in P > 1.5ng/ml). At ovulation trigger, the incidence of EP ranged from 3.1% to 65%. (33% - 52.2% in P > 0.9ng/ml; 20% - 65% in P > 1.0ng/ml; 13.9% - 31.2% in P > 1.1ng/ml; 17.2% - 54.7% in P > 1.2ng/ml; 21.5% - 32.7% in P > 1.3ng/ml; 4.6% - 44.2% in P > 1.5ng/ml; 7% - 20.1% in P > 1.7ng/ml; 11.9% in P > 1.9ng/ml and 3.1% - 33.6% in P > 2.0ng/ml). The incidence of EP at egg collection ranged from 27.3% to 55.4%. (55.4% in P > 2.0ng/ml; 39.4% in P > 11.7ng/ml and 27.3% in P > 12ng/ml). In frozen cycles, the incidence of elevated progesterone was 68.7% at ovulation trigger in modified natural cycle FET (NC-FET). The incidence of EP before ovulation in NC-FET was 24.4% (P > 1.0ng/ml) and 26.4% (P > 1.57ng/ml). Limitations, reasons for caution Observational studies were included in this systematic review with wide variation of progesterone thresholds and timing that increases the clinical heterogeneity. The incidence of EP varies widely among the included studies. The immunoassays used for progesterone measurement were not standardised with questionable accuracy at low levels. Wider implications of the findings The incidence of EP varies significantly amongst the studies. We observed similar wide variation of EP incidence at low and high progesterone threshold level. The accuracy of immunoassays on progesterone concentrations <2.5ng/ml has been questioned and the non-standardized timing of progesterone measurement challenges the reliability of the measured progesterone result. Trial registration number not applicable

Folliculogenesis and steroidogenesis alterations after chronic exposure to a human-relevant mixture of environmental toxicants spare the ovarian reserve in the rabbit model

BackgroundIndustrial progress has led to the omnipresence of chemicals in the environment of the general population, including reproductive-aged and pregnant women. The reproductive function of females is a well-known target of endocrine-disrupting chemicals. This function holds biological processes that are decisive for the fertility of women themselves and for the health of future generations. However, insufficient research has evaluated the risk of combined mixtures on this function. This study aimed to assess the direct impacts of a realistic exposure to eight combined environmental toxicants on the critical process of folliculogenesis.MethodsFemale rabbits were exposed daily and orally to either a mixture of eight environmental toxicants (F group) or the solvent mixture (NE group, control) from 2 to 19 weeks of age. The doses were computed from previous toxicokinetic data to reproduce steady-state serum concentrations in rabbits in the range of those encountered in pregnant women. Ovarian function was evaluated through macroscopic and histological analysis of the ovaries, serum hormonal assays and analysis of the expression of steroidogenic enzymes. Cellular dynamics in the ovary were further investigated with Ki67 staining and TUNEL assays.ResultsF rabbits grew similarly as NE rabbits but exhibited higher total and high-density lipoprotein (HDL) cholesterol levels in adulthood. They also presented a significantly elevated serum testosterone concentrations, while estradiol, progesterone, AMH and DHEA levels remained unaffected. The measurement of gonadotropins, androstenedione, pregnenolone and estrone levels yielded values below the limit of quantification. Among the 7 steroidogenic enzymes tested, an isolated higher expression of Cyp19a1 was measured in F rabbits ovaries. Those ovaries presented a significantly greater density/number of antral and atretic follicles and larger antral follicles without any changes in cellular proliferation or DNA fragmentation. No difference was found regarding the count of other follicle stages notably the primordial stage, the corpora lutea or AMH serum levels.ConclusionFolliculogenesis and steroidogenesis seem to be subtly altered by exposure to a human-like mixture of environmental toxicants. The antral follicle growth appears promoted by the mixture of chemicals both in their number and size, potentially explaining the increase in atretic antral follicles. Reassuringly, the ovarian reserve estimated through primordial follicles number/density and AMH is spared from any alteration. The consequences of these changes on fertility and progeny health have yet to be investigated.

FRI384 Sterilization Of Female Dogs By A Neonatal Treatment Of Estrogen-Releasing Implants

Abstract Disclosure: C. Park: None. P. Lin: None. S. Zhou: None. R.A. Hess: None. C. Ko: None. Hypothalamic Kisspeptin neurons that control the GnRH neuron activity are known to be a core regulator of the reproductive axis. Loss of the Kisspeptin-1 gene in these neurons results in infertility. Recent reports showed that inhibition of Kisspeptin neuron development by daily injection of 17β-estradiol benzoate (EB, 0.5 mg/day), a long-lasting estrogen, to neonatal female rats for 10 days causes sterility. We aim to develop a single injection solution for achieving sterilization in female dogs by targeting developing Kisspeptin neurons. In this study, we tested a working hypothesis that, in female dogs, extended exposure to EB for 2 weeks after birth by slow-releasing implants will result in sterility. This hypothesis was proposed based on our proof-of-concept study in which neonatal treatment of female rats with implants that contained 0.3 mg EB decreased the number of Kisspeptin neurons, and made them infertile permanently. To test the hypothesis in dogs, pellets that contained 0.5 mg (n=3) or 1.0 mg EB (n=2) and were designed to release EB for 2-3 weeks were subcutaneously implanted in female beagle dogs on postnatal day 6 (PND6). Intact dogs (n=4) were used as controls. The indices for female fertility (estrous cyclicity, ovulation) were assessed by daily vulva examination (swelling, discharge) and monthly measurement of serum progesterone levels. In addition, when a dog exhibited signs of estrus, the dog was bred with fertility-proven male dogs. During the 3 years of the study period, control dogs showed a minimum of 3 reproductive cycles (estrus, ovulation), and successfully produced puppies each time. Treated dogs displayed dose-dependent fertility outcomes; none of the EB 1.0 mg-treated dogs exhibited estrus nor had elevated serum progesterone levels, while two dogs out of the three EB 0.5 mg-treated dogs remained infertile. The other dog of the EB 0.5 mg group showed estrus signs during the study period and produced a litter when bred with a proven male. In the second experiment, implants that contained 0.5 mg (n=2) or 1.0 mg EB (n=3) were injected into female puppies on PND1. They are currently 1-1.2 years of age, and none have shown estrus/ovulation signs so far. Our study demonstrates that neonatal treatment of EB by slow-releasing implants sterilizes female beagle dogs. During the study period, the complete blood cell count and serum chemistry of treated dogs were within the normal range. This study shows that the non-surgical solution using EB implants is a promising approach to replace the current practice of sterilizing pets with surgical methods. Supported by NSF SBIR grant (#2052603) Presentation: Friday, June 16, 2023

P-667 The attenuated response to elevated serum progesterone levels on pregnancy outcomes of fresh IVF/ICSI cycles in older patients

Abstract Study question Whether there are different impacts of serum progesterone (P) levels on the day of trigger on pregnancy outcomes in patients of different age groups. Summary answer Our results suggest that the pregnancy outcome was less sensitive to progesterone elevation in older patients. What is known already It is now generally believed that the P elevation at the late follicular phase reduces the pregnancy rate by opening the implantation window prematurely and disrupt the embryo-endometrial synchrony. Previous studies further investigated the effect of developmental stages of embryo, number of oocytes retrieved, cause of infertility and different stimulation regimens on the association of P level and the likelihood of pregnancy. However, the study of maternal age on such association is scarce. A previous study found that LH receptor (LHCGR) and progesterone receptor (PGR) were up regulated in advanced age group. Study design, size, duration This was a non-interventional, retrospective cohort study of patients undergoing routine practice in a university-based fertility center between 2015 and 2021. One thousand five hundred and seventeen IVF/ICSI cycles among 1305 patients were included in the analysis. Participants/materials, setting, methods Fresh ET using autologous oocytes with serum P level measured on the day of hCG administration were included. Exclusion criteria include cycles without P measurement, using frozen oocytes, PGT cycles, freeze-all cycles, oocyte donation and no embryo transfer cycles. The patient age was divided into 3 groups: group 1 included patients at age 35 or below, group 2 included patients at age 36 to 39, group 3 included those at 40 years old or above. Main results and the role of chance We performed logistic regression analysis for each group to determine the contributing factors to the live birth outcome. In group 1, age, serum P level, number of usable embryos, embryo grading and endometrial thickness were found to be significantly associated with live birth. In group 2, age and number of usable embryos were significantly associated with live birth. However, total dose of gonadotropin was found to be the sole contributing factor to live birth in group 3. We also divided the P levels into low (≤0.50 ng/mL), median (0.51-1.00 ng/mL) and high (>1.00 ng/mL) and compared the live birth rate in each age groups. The live birth rate was found to be significantly reduced with increasing P levels (40.3% vs. 35.5% vs. 23.6%, p = 0.005). No significant difference in live birth rate between different P levels was found in age group 2 and 3. Limitations, reasons for caution The limitation of the study is its retrospective nature which may have led to bias in the interpretation of the data. There were also significantly higher proportion of cleavage-stage embryo to blastocyst transfer in the older group. Wider implications of the findings The negative association of premature progesterone rise at pre-ovulatory phase to the IVF fresh ET outcome was more prominent for younger patients. This may be one of the contributing factors that accounts for the great heterogeneity in the cut-off of P and its predicting power in different studies. Trial registration number Not applicable

Elevated serum progesterone levels before frozen embryo transfer do not negatively impact reproductive outcomes: a large retrospective cohort study

To evaluate whether patients with high-serum progesterone levels before frozen embryo transfer (FET) under hormonal replacement therapy present with worse reproductive outcomes. A cohort retrospective study. A university-affiliated fertility center. A total of 3,183 FET cycles in patients receiving hormonal replacement therapy between March 2009 and December 2020 were included. The luteal phase was covered with 200 mg per 8 hours of vaginal micronized progesterone either alone or in combination with a daily subcutaneous injection of 25 mg of progesterone. A total of 1,360 cycles corresponded to frozen homologous embryo transfer (ET) (hom-FET), 1,024 were euploid ET (eu-FET) after preimplantation genetic testing for aneuploidies, and 799 cycles were frozen heterologous ET (het-FET). All patients had adequate serum progesterone levels (≥10.6 ng/mL) before the procedure. Frozen embryo transfer cycles. Clinical pregnancy, miscarriage, and live birth rates (LBRs). Median (P25; P75) serum progesterone level before FET was 14.39 (12.43-17.49) ng/mL. Progesterone levels were significantly higher in the group under vaginal plus subcutaneous progesterone (15.96 [13.74-21.60] vs. 14.09 [12.19-16.95]). No differences in clinical pregnancy, miscarriage, and LBR were observed based on the use of vaginal or vaginal plus subcutaneous progesterone for each of the groups (hom-FET, eu-FET, and het-FET). Live birth rates were comparable among patients in the highest centile of serum progesterone levels (≥p90) (22.33 ng/mL) and the rest of the patients (p<90) (43.9% vs. 41.3%). Patients with progesterone levels ≥p90 presented lower body mass index than those in the lower centiles (<p90) (22.62 ± 3.82 vs. 23.32 ± 4.06). After dividing patients into deciles based on serum progesterone levels, no differences in LBRs were observed among the groups. No association was observed using a generalized additive model between progesterone levels and LBR. A multivariable logistic regression adjusted by oocyte age, type of treatment, body mass index, type of luteal phase support, and the number of embryos transferred was applied for centile 90 and centile 95 of progesterone and showed that serum progesterone in their highest levels did not negatively impact LBR. Elevated serum progesterone levels before FET do not impair reproductive outcomes in patients receiving artificially prepared cycles with vaginal or vaginal plus subcutaneous progesterone.

Adverse impact of elevated serum progesterone and luteinizing hormone levels on the hCG trigger day on clinical pregnancy outcomes of modified natural frozen-thawed embryo transfer cycles.

The relationship between serum progesterone (P) and luteinizing hormone (LH) levels on the human chorionic gonadotropin (hCG) trigger day and the clinical pregnancy outcomes in modified natural frozen-thawed embryo transfer (mNC-FET) cycles are controversial. This was a retrospective study of 788 mNC-FET cycles. A smooth fitting curve and threshold effect analysis was performed to identify the effect of serum P and LH levels measured on the hCG day on the clinical pregnancy rate (CPR) and live birth rate (LBR) of mNC-FET cycles. The CPR and LBR decreased significantly when the LH level on the hCG day was greater than or equal to 32 IU/L. Further subgroup analysis showed that the CPR decreased significantly when the P level on the hCG day was equal to or greater than 1 ng/mL. When the P level was lower (< 1 ng/mL), the patients with an LH level greater than or equal to 32 IU/L had reduced CPR and LBR in mNC-FET cycles. Applying the hCG trigger on a day with a higher P level (≥ 1 ng/mL) leads to a decreased CPR and LBR. hCG administration with a higher LH level (≥ 32 IU/L) also leads to a decreased CPR and LBR in mNC-FET cycles when the P level is less than 1 ng/mL.

Elevated serum progesterone levels on the hCG trigger day have a negative impact on the live birth rate in the first fresh IVF-ET cycle

To investigate whether there is a relationship between elevated serum progesterone (PROG) on the hCG trigger day and the live birth rate (LBR) in IVF/ICSI cycles, the retrospective analysis was carried out from the patients undergoing the first ART cycles throughout 2016. The PROG levels were measured on the hCG trigger day. The LBR, clinical pregnancy rate (CPR), implantation rate (IR) and other parameter rate values were compared among the three different PROG elevations. A total of 2550 IVF/ICSI cycles fulfilling all the inclusion and exclusion criteria were selected. Finally, three groups [PROG <0.40 ng/mL, 0.40 ≤ PROG < 1.5 ng/mL, PROG ≥ 1.5 ng/mL] were categorised based on the serum PROG levels. LBR, CPR and IR declined as the PROG value increased, while there was no difference in the embryo utilisation rates. Serum PROG levels on the day of hCG administration were negatively associated with the LBR, even in ETs with a good prognosis. Impact Statement What is already known on this subject? The clinical effects of PROG are still controversial. Some studies have confirmed that there was not too much association between premature elevation of PROG and live birth, some are still convincing that there is a negative correlation and will result in ART cycles of pregnancy and LBR reduction. What do the results of this study add? Our data substantiated that the high serum PROG level had the lowest LBR, IR and CPR, but the embryo utilisation rate may not have too much to do with the elevated PROG. What are the implications of these findings for clinical practice and/or further research? This study further strengthens the negative impact of elevated PROG levels on pregnancy outcomes, and suggests that frozen thawed embryo transfer appears to be a reasonable and advantageous approach for this subset of patients.

Transcutaneous electrical acupoint stimulation improves endometrial receptivity resulting in improved IVF-ET pregnancy outcomes in older women: a multicenter, randomized, controlled clinical trial

ObjectiveTo examine the effects and mechanisms of transcutaneous electrical acupoint stimulation (TEAS) on pregnancy outcomes in women undergoing in vitro fertilization (IVF)-embryo transfer (ET).Design, setting, and participantsThis efficacy study was a multicenter, randomized, controlled clinical trial (RCT) in women receiving IVF-ET. The mechanistic study was a single-center RCT.InterventionsThe participants received TEAS vs. no TEAS treatment.Main outcome measuresIn the efficacy study, the primary outcomes were the rates of clinical pregnancy, embryo implantation, and live birth. In the mechanistic study, sex hormones and endometrial protein expression were examined.ResultsUltimately, 739 participants were enrolled (367 and 372 in the TEAS and control groups, respectively). The clinical pregnancy rate was higher in the TEAS group than in the controls (55.1% vs. 46.7%, P = 0.03). There were no significant differences in embryo implantation, biochemical pregnancy, and live birth rates between the two groups (all P > 0.05) in the study population. In women > 35 years, the clinical pregnancy rates, embryo implantation rates and live birth rates in the TEAS and control groups were 48.9% vs. 23.7% (P = 0.004),30.8 vs. 13.9% (P = 0.001) and 34.0% vs. 19.7% (P = 0.06) respectively. In the mechanistic study with 120 participants, on the theoretical embryo implantation day, better developed endometrial pinopodes, elevated endometrial integrin α1β1/αVβ3, leukemia inhibitory factor, and elevated serum progesterone levels were found in the TEAS group compared with controls.ConclusionTEAS significantly improved the clinical pregnancy rate in women undergoing IVF-ET, especially in women of older age. It might be due to improved endometrial receptivity.Trial registrationChiCTR-TRC-13003950.

Adverse impact of elevated progesterone levels on human chorionic gonadotropin trigger day on blastocyst transfer outcomes in gonadotropin-releasing hormone agonist cycles

ObjectiveDose an elevated serum progesterone (P) level on the human chorionic gonadotropin (hCG) trigger day have a negative effect on clinical pregnancy outcomes for embryos transferred at different stages of development in long-acting gonadotropin-releasing hormone agonist (GnRHa) in vitro fertilization-embryo transfer (IVF-ET) cycles? Study designThis was a noninterventional, retrospective, observational, single-centre cohort study. A total of 1951 patients received long-acting GnRHa for pituitary downregulation in IVF-ET cycles at Nanjing Drum Tower Hospital from January 2018 to December 2020. The serum P levels on the day of hCG administration were measured, together with other cycle parameters, to explore the relationship between P levels and the clinical pregnancy rate (CPR) of different embryos transferred. ResultsWhen the serum P level on the hCG day was higher than 1.5 ng/mL, the CPR did not decrease significantly. There was no correlation between the CPR of cleavage-stage embryo transfer and the serum P level on the hCG day. In addition, the interaction analysis suggested that the CPR of patients undergoing blastocyst transfer decreased as serum P levels on the hCG day increased. Progesterone levels on the day of hCG administration were closely related to the CPR of blastocyst transfer rather than cleavage-stage embryo transfer. ConclusionAn increased serum P level on the day of hCG administration did not affect the CPR of cleavage-stage embryo transfer, but it reduced the CPR of blastocyst transfer cycles.

O-241 Elevated serum progesterone levels before frozen embryo transfer do not negatively impact reproductive outcomes: a large retrospective cohort study

Abstract Study question Do patients with high serum progesterone levels before frozen embryo transfer (FET) under hormonal replacement therapy (HRT) present worse reproductive outcomes? Summary answer Elevated serum progesterone levels before FET in artificially prepared cycles with vaginal or vaginal plus subcutaneous progesterone do not impair reproductive outcomes. What is known already Low serum progesterone levels before FET do negatively affect reproductive outcomes in terms of live birth rate. However, there is not robust data regarding the impact of high serum progesterone levels in the luteal phase of patients who undergo HRT for FET. Study design, size, duration Retrospective cohort study of 3183 blastocyst FET cycles under HRT performed in a university-affiliated fertility centre between March 2009 and December 2020. All the cycles presented adequate serum progesterone levels before FET (≥10.6 ng/ml). A total of 1360 cycles corresponded to frozen homologous embryo transfer (ET) (hom-FET), 1024 were euploid ET (eu-FET) after preimplantational genetic testing for aneuploidies (PGT-A), and 799 cycles were frozen heterologous ET (het-FET). The primary objective was live birth rate (LBR). Participants/materials, setting, methods Standard HRT was used. Luteal phase was covered with vaginal progesterone 200 mg/8h, or vaginal plus a daily subcutaneous injection of progesterone (25 mg). Serum progesterone levels were measured the day before FET. Elevated progesterone levels were considered in the 90th and 95th centiles. A generalized additive model (GAM) was performed to study the functional relationships between progesterone and LBR. A multivariable logistic regression was used to evaluate the effect of high progesterone over LBR. Main results and the role of chance Mean serum progesterone level before FET was 16.77±8.43 ng/ml. Progesterone levels were significantly higher in the group under vaginal plus subcutaneous progesterone (21.87±14.17 vs. 15.56±5.72, p &amp;lt; 0.001). No differences in clinical pregnancy, miscarriage and LBR were found according to the use of vaginal or vaginal plus subcutaneous progesterone for each of the groups (hom-FET, eu-FET and het-FET). Live birth rates were comparable among patients in the highest centile of serum progesterone levels (≥p90, ≥22.33 ng/ml) and the rest of patients (p &amp;lt; 90) (43.9 vs 41.3%; p = 0.381). Patients with progesterone levels ≥p90 presented lower BMI compared to those in the lower centiles (&amp;lt;p90) (22.62±3.82 vs. 23.32±4.06; P = 0.009). After dividing patients in deciles according to serum progesterone levels before, no differences in LBR were observed among groups (P = 0.938). No association was observed with GAM model between progesterone levels and LBR. A multivariable logistic regression adjusted by oocyte age, type of treatment and number of embryos transferred was applied for centile 90 and centile 95 of progesterone, and showed that serum progesterone in their highest levels did not negatively impact LBR. Limitations, reasons for caution The main limitation of this study is its retrospective design. The results only apply for patients under HRT with vaginal micronized progesterone alone or plus subcutaneous progesterone. Progesterone determination was measured before blastocyst FET. Extrapolation to other HRT protocols or timings of progesterone measurement needs to be validated. Wider implications of the findings The results of this study suggest that once a threshold of serum progesterone before FET is achieved, progesterone levels are not predictive of the clinical outcome. Actually, LBR are not negatively affected when progesterone levels are found in their highest centiles after luteal phase rescue with vaginal plus subcutaneous progesterone. Trial registration number Not applicable

Anti-inflammatory and hemostatic effects of total extract, saponins, and flavonoids of Clinopodium chinense in female rats with abnormal uterine bleeding and mechanism

This study aims to explore the anti-inflammatory and hemostatic effects of the total extract of Clinopodium chinense(TEC), total saponins of C. chinense(TSC), and total flavonoids of C. chinense(TFC) in female rats with abnormal uterine bleeding(AUB), and the possible mechanism. Mifepristone(i.g., 12.4 mg·kg~(-1)) and misoprostol(i.g., 130 μg·kg~(-1)) were used to induce AUB in SD female rats conceiving on the same day. Then the AUB rats were randomized into model group, TEC group, TSC group, TFC group, Yimucao Granules(LG) group, and estradiol valerate(EV) group, with 8 rats in each group. Another 8 non-pregnant female rats were selected as normal group. During the experiment, each group was given the corresponding drug by gavage once a day for 7 days. After the administration, blood and uterine tissue were collected. The uterine bleeding volume was measured by ultraviolet spectrophotometry and the pathological changes of endometrium were observed based on hematoxylin-eosin(HE) staining. In addition, the microvessel density of endometrium was determined by immunohistochemistry, and the content of thromboxane B2(TXB2), 6-keto-PGF_(1α), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in plasma and levels of lutenizing hormone(LH), follicle stimulating hormone(FSH), estradiol(E_2), and progesterone in serum were detected by enzyme-linked immunosorbent assay(ELISA). The mRNA and protein expression of estrogenreceptor α(ERα), progesterone receptor(PR), matrix metalloproteinase(MMP)-2, MMP-9, and vascular endothelial growth factor(VEGF) in uterine tissue was determined by Western blot. Compared with the model group, TEC, TSC, and TFC can reduce uterine bleeding volume, alleviate the pathological damage of endometrium, and increase the microvessel density in endometrium. Moreover, TEC and TSC can significantly raise plasma TXB2 level and ratio of TXB2 to 6-keto-PGF_(1α), and TEC and TFC can significantly reduce the levels of IL-6 and TNF-α. In addition, TEC significantly elevated serum progesterone level and TFC significantly increased serum levels of E_2, FSH, and LH. TSC can significantly raise serum progesterone and FSH levels. In addition, TEC can significantly down-regulate the protein expression of PR, MMP-2, and VEGF and TSC significantly reduced the expression of MMP-9. TFC significantly decreased the expression of PR, MMP-9, and VEGF, and up-regulated the expression of ERα. In conclusion, TEC, TSC, and TFC all show therapeutic effects on AUB, particularly TEC. TSC exerts the effects by enhancing the coagulation function and promoting endometrial repair, and TFC by regulating estrogen levels and reducing inflammatory response. This study reveals the mechanism of C. chinense against AUB and also explains the holistic characteristics of Chinese medicine.

Serum progesterone on the day of human chorionic gonadotropin (hCG) trigger as a predictor of in-vitro fertilization (IVF) outcome - a retrospective analysis of seven years.

To investigate the relationship between progesterone (P4) levels on the day of hCG trigger and IVF outcomes. This is a retrospective analysis of IVF cycles from January-2013 to December-2019 from a single center. Women (21-39 years) submitted to IVF treatment for various infertility factors were included, while donor oocyte cycles and cancelled cycles were excluded from the study. The primary outcome measure was live birth rate. A total of 2149 cycles were analyzed. Of these, 223 (10.38%) were in the low P4 group (<0.5 ng/ml), 1163 (54.12%) in the normal P4 group (0.5-1.5 ng/ml), and 763 (35.50%) in the high P4 group (>1.5ng/ml). The groups were comparable with respect to age, factor of infertility and baseline AMH. The antagonist protocol was significantly more prescribed to the high P4 group (p<0.001). Live birth rates were 14.4%, 21.6%, and 21% (p<0.001), respectively, in three groups. Univariate analysis found that total cetrotide dose, total number of retrieved and fertilized oocytes, total number of embryos formed, transferred, and vitrified, and P4 on the day of hCG (p<0.001) were statistically significant after adjusting for age and BMI. In multivariate logistic regression after adjusting for age and BMI, only high P4 (aOR:0.60; p<0.001), total cetrotide dose (aOR: 0.82; p<0.001), and total utilizable embryos (aOR:1.11; p=0.029) were statistically significant. Having an elevated serum progesterone level on the day of hCG trigger was associated with lower pregnancy rates, but this is still not a robust marker to predict live births. More good quality evidence is needed.

P–660 Impact of elevated late-follicular phase serum estrogen and progesterone levels on blastocyst utilization and cumulative live birth rates in freeze-all cycles

Abstract Study question Does elevated late-follicular phase estrogen and progesterone levels have an impact on blastocyst utilization and/or cumulative live birth rates in freeze-all cycles? Summary answer High estrogen or progesterone on the day of ovulation trigger is associated with poor blastocyst utilization but comparable cumulative live birth rates in freeze-all cycles. What is known already Several studies suggest impaired clinical outcome in cycles with high estrogen (&amp;gt;3500 pg/ml) or progesterone (&amp;gt;1.5 ng/ml) levels. However, these data were derived from cycles where top-quality embryo(s) were transferred in the fresh cycle and surplus embryos were frozen. These findings might be confounded by alterations in endometrial receptivity. Freeze-all cycles might provide a better model to assess the impact of high late-follicular estrogen or progesterone levels on laboratory and clinical outcome. Study design, size, duration We performed a retrospective cohort study of all IVF cycles (n = 712) between 2016 and 2018 where the entire cohort of embryos was cryopreserved at the blastocyst stage. After excluding cases with &amp;lt;4 oocytes or preimplantation genetic test, the study group comprised 459 women who had 699 frozen-thawed embryo transfer cycles. Participants/materials, setting, methods Women were classified into four groups by the indication for freeze-all strategy as elevated progesterone (high P, n = 61), high estrogen (high E, n = 224), elective freezing (elective, n = 114) and tubal-endometrial pathologies (TEP, n = 60). The primary outcome was the cumulative live birth rate in subsequent thaw-transfer cycles and the secondary outcome was the blastocyst utilization rate. Groups were compared using ANOVA and Cox regression analyses to adjust for confounding variables. Main results and the role of chance The mean age of the study group was 32.8 ± 5.3 years, total number of oocytes and cryopreserved blastocysts were 15.0±7.6 and 4.2±3.0, respectively. The high-E group was younger (31.5 ± 5.2 years) and had higher peak E2 levels (4078.9 ± 588.4 pg/ml), number of oocytes (19.7 ± 7.0), cryopreserved embryos (5.3 ± 3.3) and transfer cycles (2.3 ± 1.4) than the other groups. Blastocyst utilization rate was significantly lower (40.4%) compared to elective freezing (53.6%) and TEP groups (55.7%) (both p = 0.001). The high-P group had higher peak progesterone levels (2.1 ± 0.5 ng/ml, p = 0.001), number of oocytes (14.0 ± 5.2) and frozen embryos (4.1 ± 3.5) compared to elective and TEP groups (both p = 0.04). Blastocyst utilization rate was lower (45.7%) than elective freezing and TEP groups but the difference lacked statistical significance (p = 0.33 and p = 0.21, respectively). Cumulative live birth rates were 42.6% in high-P, 59.8% in high-E, 44.7% in elective freezing and 46.7% in TEP groups. Significant predictors of cumulative live birth were female age (aHR: 0.97, 95%CI:0.95–0.99, p = 0.02) and number of frozen blastocysts (aHR:1.05, 95%CI:1.01–1.10), p = 0.02). When adjusted for these confounders, the cumulative live birth rate was not associated with high-E (aHR: 0.86, 95%CI:0.56–1.31) or high-P (aHR: 0.76,95%CI:0.44–1.32). Limitations, reasons for caution This was a retrospective study with small sample size performed at a single fertility center, which may limit the generalizability of our findings. Wider implications of the findings: While lower blastocyst utilization rates are observed in women high late-follicular estradiol or progesterone levels, cumulative live birth rates in subsequent thaw-transfer cycles were not impaired. However, unfavorable outcome parameters observed in women with elevated progesterone deserve further research. Trial registration number Not applicable

Ovarian Hyperstimulation Syndrome Is Associated with a High Secondary Sex Ratio in Fresh IVF Cycles with Cleavage-Stage Embryo Transfer: Results for a Cohort Study.

The sex ratio at birth is defined as the secondary sex ratio (SSR). Ovarian hyperstimulation syndrome (OHSS) is a serious and iatrogenic complication associated with controlled ovarian stimulation (COS) during assisted reproductive technology (ART) treatments. It has been hypothesized that the human SSR is partially controlled by parental hormone levels around the time of conception. Given the aberrant hormonal profiles observed in patients with OHSS, this retrospective study was designed to evaluate the impact of OHSS on the SSR. In this study, all included patients were divided into 3 groups: non-OHSS (n=2777), mild OHSS (n=644), and moderate OHSS (n=334). Our results showed that the overall SSR for the study population was 1.033. The SSR was significantly increased in patients with moderate OHSS (1.336) compared to non-OHSS patients (1.002) (p=0.048). Subgroup analyses showed that increases in the SSR in patients with moderate OHSS were observed in the IVF group (1.323 vs 1.052; p=0.043), but not in the ICSI groups (1.021 vs 0.866; p=0.732). In addition, the elevated serum estradiol (E2) and progesterone (P4) levels in OHSS patients were not associated with SSR. In this study, for the first time, we report that a high SSR is associated with OHSS in patients who received fresh IVF treatments. The increases in SSR in OHSS patients are not attributed to the high serum E2 and P4 levels. Our findings may make both ART clinicians and patients more aware of the influences of ART treatments on the SSR and allow clinicians to counsel patients more appropriately.

RATIO OF HUMAN MENOPAUSAL GONADOTROPIN (HMG) TO RECOMBINANT FOLLICLE STIMULATING HORMONE (RFSH) DOSAGE PREDICTS PREMATURE PROGESTERONE ELEVATION AND LIVE BIRTH RATE IN FRESH IVF CYCLES

Premature progesterone rise during IVF stimulation reduces endometrial receptivity and is associated with decreased pregnancy rates following embryo transfer (ET). To our knowledge, no data exists regarding the appropriate ratio of hMG (which has 1:1 LH:FSH activity) to rFSH (which has FSH activity alone) to improve pregnancy outcomes. Our objective is to determine if an optimal ratio of hMG:rFSH can be identified that best predicts a reduced risk of elevated serum progesterone levels and change in progesterone (ΔP) during IVF stimulation, and to determine whether this ratio can also predict pregnancy outcomes following fresh ET.

Stop GnRH-Agonist Combined with Multiple-Dose GnRH-Antagonist for Patients with Elevated Peak Serum Progesterone Levels Undergoing Ovarian Stimulation for IVF: A Proof of Concept

Aim: The aim of the study was to examine whether the Stop GnRH-agonist combined with multiple-dose GnRH-antagonist protocol may overcome progesterone elevation during the late follicular phase. Patients and Methods: A cohort historical, proof of concept study consisting of 11 patients with progesterone elevation (>3.1 nmol/L) during conventional IVF/intracytoplasmic sperm injection (ICSI), who underwent a subsequent Stop GnRH-agonist combined with multiple-dose GnRH-antagonist ovarian stimulation (OS) protocol, within 3 months of the previous failed conventional IVF/ICSI cycle. Results: The Stop GnRH-agonist combined with multiple-dose GnRH-antagonist COH protocol revealed significantly lower peak progesterone levels, with significantly higher numbers of follicles >13 mm in diameter on the day of hCG administration, oocytes retrieved, mature oocytes, and top-quality embryos, with an acceptable clinical pregnancy rate (18.2%). Conclusions: The combined Stop GnRH-ag/GnRH-ant OS protocol is a valuable tool in the armamentarium for treating patients with progesterone elevation during the late follicular phase. Further large prospective studies are needed to validate our observation and to characterize the appropriate patients’ subgroup, which might benefit from the combined Stop GnRH-ag/GnRH-ant COH protocol.

Suboptimal response to GnRH-agonist trigger during oocyte cryopreservation: a case series

BackgroundRandom-start, controlled ovarian stimulation (COS) has advanced the field of fertility preservation, allowing patients to expedite fertility treatment and avoid further delays to their cancer therapy. This novel approach allows patients to initiate ovarian stimulation at any point, regardless of where they are in their menstrual cycle. Luteal-phase start (LPS) protocols describe treatment cycles where COS is initiated during the luteal-phase of the menstrual cycle. LPS protocols have not been studied or optimized to the same degree as conventional, early-follicular COS. Particularly, there is a paucity of evidence evaluating treatment outcomes using different trigger medications in LPS protocols. The present study aims to evaluate the efficacy of using a GnRH agonist (GnRH-a) trigger in patients undergoing oocyte cryopreservation in LPS protocols.MethodsThis descriptive case series describes two patients, recently diagnosed with cancer, who underwent oocyte cryopreservation using an LPS protocol and a GnRH-a trigger at a university-affiliated, academic center.ResultsThe patients described in our case series both failed to adequately respond to a GnRH-a trigger, based on their serum levels of luteinizing hormone (LH) and progesterone 12 h after their GnRH-a trigger. They both required a single rescue dose of human chorionic gonadotropin (hCG).ConclusionsThese findings highlight the potential risk of a suboptimal response to a GnRH-a trigger in patients undergoing LPS, controlled ovarian stimulation for oocyte cryopreservation. This risk might be attributed to the downregulation of GnRH receptors by elevated serum progesterone levels during the luteal phase. Currently, there is insufficient evidence to recommend for or against the use of a GnRH-a trigger during LPS controlled ovarian stimulation. This case series offers a number of management strategies to mitigate this risk and emphasizes the need for further research in this area.

Progesterone in Assisted Reproduction: Classification, Pharmacology and Its Clinical Coorelation: A Commentary

The modulating effects of progesterone on endometrium structure and function are the basis for successful outcome in reproductive treatments. Considering this, progesterone has a big role in treatment of infertility and supporting the ongoing pregnancy. In this review article we have attempted to review different forms of progesterone, their metabolism in the body and the role played by exogenous as well as endogenous progesterone in assisted reproductive technologies (ART). Elevated serum progesterone levels at the end of the follicular phase in controlled ovarian stimulation (COS) leads to a poorer ongoing pregnancy rate in IVF cycles due to reduced endometrial receptivity.

Sweet taste receptor agonists alter ovarian functions and ovarian cycles in aged mice

Saccharine sodium and rebaudioside A are low-calorie sweeteners, and the biologic effects of these sweeteners in rat ovaries are related to the activity of sweet taste receptors. Data on the impact and regulatory mechanisms underlying such sweeteners on the reproduction of aged animals are currently lacking. In the present study we assessed how the consumption of sweeteners affects the ovarian cycle, ovulation, biochemical indices, and other biologic functions. Thirty-six 1-year-old mice were randomly divided into 3 groups: a control (C) group receiving regular water, a saccharin sodium group receiving a 7.5 mM solution, and the rebaudioside A group receiving a 2.5 mM solution for 30 days. We observed no significant changes in body weights in any group. However, uterine weight in the rebaudioside A group significantly increased in diestrus, and we recorded a significant increase in the percentage of abnormal estrous cycles and the number of corpora lutea in the treatment groups. TUNEL staining and Immunoreactivity for the apoptosis-inducing factor (AIF) confirmed apoptosis in granulosa cells, oocyte, and corpus luteum. Serum glucose increased significantly in both treatment groups and there was a significant increase in cholesterol in the rebaudioside A group. Furthermore, the saccharin sodium-treated group exhibited elevated serum progesterone levels compared with the other groups. In conclusion, sweeteners manifested deleterious effects on reproductive indices in aged mice.