Serum E2 Research Articles

Mechanism study of YangJing ZhongYu decoction on regulating mitochondrial dynamics of ovarian granular cells and improving diminished ovarian reserve

ObjectiveDiminished ovarian reserve (DOR) encompasses both reproductive and endocrine disorders, resulting in a decline in female fertility. This paper explored the mechanism of Yangjing Zhongyu Decoction (YJZYD) regulating mitochondrial dynamics of ovarian granulosa cells (GCs) to improve DOR.MethodsDOR patients were treated with YJZYD, with ovarian volume (OV), antral follicle count (AFC), and endometrial thickness (EMT) detected. C57BL/6 female mice were treated by cyclophosphamide (Cy) intraperitoneal injection and YJZYD solution daily gavage, with serum anti-Mullerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) levels determined. Ovarian GCs (KGN) were interfered with 4-Hydroperoxy-Cyclophosphamide (4-HC) and treated with the MAPK/ERK pathway inhibitor or activator.ResultsDOR patients showed increased levels of serum AMH, E2, OV, AFC and EMT, while reduced FSH and LH levels after YJZYD treatment. After Cy induction, DOR mice exhibited irregular estrous cycles, diminished serum AMH and E2 levels, elevated FSH and LH levels, reduced follicle number and atresia follicle number, disorderly arranged GCs, and severe interstitial fibrosis. After 4-HC treatment, KGN proliferation and Bcl-2, MFN1, and MFN2 were suppressed, while apoptotic rate, Bax, Cleaved-caspase-3, and p-Drp1 (Ser616) levels, and mitochondrial fission and quantity increased. YJZYD promoted 4-HC-treated KGN proliferation, boosted mitochondrial fusion, and inhibited apoptosis and mitochondrial fission via the MAPK/ERK pathway.ConclusionYJZYD promoted ovarian GC proliferation and mitochondrial fusion, suppressed cell apoptosis and mitochondrial fission, and effectively improved DOR in mice by activating the MAPK/ERK pathway, providing a theoretical basis for the clinical application value of YJZYD in DOR treatment.

Study of hypothalamic-pituitary-testicular axis in Egyptian post-COVID-19 patients

ABSTRACT Background SARS-CoV-2, was identified as a global pandemic by the World Health Organization a few months after the first epidemic. It penetrates host cells through angiotensin-converting enzyme 2 receptors and spreads by respiratory droplets. Testes are among the several tissues that have high expression levels of ACE-2 receptors. The current study aimed to evaluate the hypothalamic-pituitary-testicular axis integrity among Egyptian post-COVID-19 patients. Methods This is a cross-sectional study that included patients with a history of COVID-19 infection, at least 3 months from the onset of COVID-19. A detailed clinical history was obtained, and a thorough examination was performed. Laboratory tests included FBG, total testosterone, free testosterone, bioactive testosterone, FSH, LH, prolactin, E2, albumin, and SHBG. Testicular ultrasonography was performed. The data were statistically analyzed. Results Eighty male patients were evaluated with a mean age of 35.53 ± 10.35 years. The mean serum total testosterone was 5.48 ± 1.65 ng/ml. Low testosterone was detected in 2.8% of the patients. High LH levels were detected in 17/80 patients (21.3%). High FSH levels were detected in 4/80 patients (5%). Six patients had elevated serum E2 level. The mean serum prolactin value was 10.90 ± 5.66 ng/ml. Fourteen patients had elevated prolactin levels. The mean serum T/LH ratio was 3.28 ± 1.52. The ratio of T/LH was decreased in eight COVID-19 patients (10%). In COVID-19 patients, a statistically significant negative correlation was found between age and free testosterone (FT) (r=-0.381, p < 0.001), bioavailable testosterone (Bio. T) (r=-0.375, p = 0.001), and androgen index (A. index) (r=-0.446, p < 0.001). Conclusion The data of the current study suggest that COVID-19 infection might deteriorate pituitary – gonadal axis. The prevalence of low testosterone in COVID-19 male survivors is low. The risk of hypogonadism is increased with advanced age. More studies are required to confirm the relationship between COVID-19 infection and male gonadal function.

Maternal dnd1 is essential for migration and maintenance of PGCs in Nile tilapia at larval stage

As a germ-cell-specific maternal mRNA and RNA binding protein, dead end (dnd1) plays an important role in migration and maintenance of primordial germ cells (PGCs), and therefore, disruption of dnd1 results in no germ cells. However, the effect and duration of maternal dnd1 on PGCs in zygote mutants have not been reported yet. In the present study, CRISPR/Cas9 was used to mutate dnd1 in tilapia. Supported by the maternal dnd1 mRNA, germ cells of the dnd1 homozygous mutants proliferated and migrated normally before 7 dpf (days post fertilization). After the degradation of maternal dnd1 mRNA, significant downregulation of piwil1, piwil2, nanos2, nanos3 and vasa mRNA, and blocked migration of germ cells were observed at 14 dpf. In dnd1 zygote mutants, the number of germ cells was decreased significantly compared with wild type (WT) at 25 dpf, and no germ cell was observed at 30 dpf. Consistently, no expression of vasa and nanos3 was detected in the adult dnd1 mutants by qPCR. Interestingly, Cyp19a1a/cyp19a1a and foxl2 were expressed in dnd1-deficient XX gonads, while Cyp11c1/cyp11c1 and dmrt1 were expressed in dnd1-deficient XY gonads as in WT gonads. Correspondingly, both serum E2 level in XX and 11-KT level in XY fish did not differ significantly between dnd1 mutant and WT fish. Taken together, the present study demonstrates that disruption of dnd1 lead to progressive loss of germ cells, while the somatic cells are not affected in tilapia. The sexual fate of gonads is determined by the genetic sex instead of germ cell number, which makes tilapia a potential surrogate model for germ cell transplantation.

Immuno-hormonal regulation of tumor proliferation in breast cancer patients

It is well known that tumor cells proliferation is regulated by sex steroid hormones, estradiol and progesterone (E2 and Pg) in breast cancer patients (BCP). Moreover, specific auto-antibodies to estrogen receptor (ERα) were detected in blood serum of BCP. Their levels positively correlated with the percentage of Ki-67 expressing breast cancer cells. We proposed that antiidiotypic auto-antibodies to E2 and Pg (IgG2-E2 and IgG2-Pg) could act as antibodies against membrane ER and progesterone receptor (PR). Our study aimed for research of IgG2-E2 and IgG2-Pg according to E2 and Pg serum levels in association with Ki-67 positive tumors in BCP. Antiidiotypic antibodies were studied in ER-positive patients with breast cancer (stage I, n = 374, and stage II-IV, n = 379,) using ELISA technique with monoclonal antibodies against E2 and Pg as adsorbed antigens. Blood serum concentrations of E2 and Pg were measured using “ImmunoEA-Estradiol” and “ImmunoEA-Progesterone” test-systems (“Immunotech”, Russia). Tumor Ki-67 was studied by standard immunohistochemical technique at the Kemerovo Oncological Hospital. Higher percentage of Ki-67 positive breast cancer cells (Ki-67 30) was increased in BCP II-IV stages compared with stage I patients (50.7% vs 29.8%, p 0,001). Such increased values were detected for the BCP with low levels of both IgG2-E2 and IgG2-Pg antibodies in the following subgroups: 1) at low serum E2 concentration of ≤ 200 pmol/L (50.9% vs 21.3%, р 0.001); 2) at the E2 exceeding 200 pmol/L (74.2% vs 34.1%, р = 0.003); 3) at the Pg levels under 600 pmol/L (60.6% vs 22.2%, р 0.001); 4) at Pg values exceeding 600 pmol/L (58.5% vs 30.8%, р = 0.02). Similar differences were not revealed between stage II-IV and stage I BCP with low levels of both IgG2-E2 and IgG2-Pg. Corresponding Ki-67 30 indices were as follows: 1) 36.9% vs 22.0% (р = 0.14); 2) 48.4% vs 34.5% (р = 0.08); 3) 34.0% vs 27.7% (р = 0.75). Significant differences were detected in BCP with Pg 600 pmol/L and high IgG2-E2 and IgG2-Pg levels only: 48.1% vs 26.6%, (р = 0.01). Hence, antiidiotypic auto-antibodies to steroid hormones may participate in regulation of tumor proliferation in BCP. Immunoassay of IgG2-E2 and IgG2-Pg may be used for prognosis of tumor proliferation upon breast cancer progression.

Efficacy analysis of letrozole combined with gonadotropin-releasing hormone antagonists in patients with high risk of ovarian hyperstimulation syndrome undergoing total embryo freezing

Objective: To investigate the clinical efficacy of letrozole combined with gonadotropin-releasing hormone antagonists (GnRH-ant) in patients at high risk of ovarian hyperstimulation syndrome (OHSS) who underwent total embryo freezing after oocyte retrieval. Methods: A retrospective analysis was conducted on 348 female patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) at the Reproductive and Genetic Hospital of the First Affiliated Hospital of Zhengzhou University between January and July 2023. Due to their high risk of OHSS, these patients canceled fresh embryo transfer and opted for total embryo freezing. Based on patients' preferences, those who received GnRH-ant and letrozole after oocyte retrieval were categorized as the intervention group (164 cases), while those who did not receive these medications were categorized as the control group (184 cases). The first luteal phase after oocyte retrieval, OHSS grading, ovarian volume, and estradiol (E2) levels were evaluated in both groups. A multivariate logistic regression model was used to analyze factors related to moderate-to-severe OHSS among patients at high risk of OHSS who underwent total embryo freezing after oocyte retrieval. Results: The age of the intervention and control groups was (29.3±3.8) and (29.4±4.1) years, respectively (P=0.821). The duration of the first luteal phase post-oocyte retrieval was shorter in the intervention group [(7.16±1.39) days] compared to that in the control group [(13.88±2.11) days] (P<0.001). The incidences of mild, moderate, and severe OHSS in the intervention group were 75.0% (123 cases), 23.8% (39 cases), and 1.2% (2 cases), respectively, whereas in the control group they were 12.5% (23 cases), 60.9% (112 cases), and 26.6% (49 cases) (P<0.001). E2 levels on the 2nd and 6th days after oocyte retrieval [M(Q1,Q3)] in the intervention group were 1 520.0 (1 213.8, 1 884.8) and 108.5 (45.6, 218.0) ng/L, respectively, which were statistically significantly lower than those in the control group [1 666.0 (508.8, 1 702.0) ng/L] and [1 761.0 (826.0, 2 546.5) ng/L] (P<0.001). The abdominal cavity effusion in the intervention group [M(Q1,Q3)] were 19.5 (0, 30) and 0 mm, statistically significantly less than those in the control group [46.0 (0, 61.0) mm] and [54.5 (0, 69.5) mm] (P<0.001). On the 6th day after oocyte retrieval, the bilateral ovarian volumes in the intervention group were smaller than those in the control group (P<0.001). Multivariate logistic regression analysis indicated that no combined treatment with letrozole and GnRH-ant was a risk factor of moderate to severe OHSS. The risk of developing moderate to severe OHSS in the control group was 35.312 times higher than that in the intervention group (OR=35.312, 95%CI: 17.488-71.300). Conclusions: The administration of letrozole combined with GnRH-ant post-oocyte retrieval in patients at high risk of OHSS can prevent the occurrence of moderate-to-severe OHSS, shorten the first luteal phase, accelerate the reduction of serum E2 levels, and promote the recovery of ovarian volume and absorption of abdominal fluid.

Characterization of sexual size dimorphism in mandarin fish and association with igfbp-5a/b regulation.

Mandarin fish (Siniperca chuatsi) is an important cultured fish in East Asia that shows sexual size dimorphism (SSD), with females growing faster than males when reaching marketable size. However, the regulatory mechanism of SSD is not clear. To characterize SSD of mandarin fish and its association with gh/igf1/igfbp-5 expression, gonadal developmental atlas of the females and the males were described, and growth parameters and serum levels of E2 and T, as well as the relative expression levels of gh, igf1, and igfbp-5a/b mRNAs, were determined. The results showed that the logistic growth equation of body mass and total length of female and male were W(♀) = 667.57/(1 + e^(4.19 - 1.24*t)), W(♂) = 582.71/(1 + e^(4.07 - 1.27*t)), L(♀) = 31.47/(1 + e^1.95 - 1.08*t)), L(♂) = 26.20/(1 + e^(2.56 - 1.5*t)). The month of inflection points for body mass for females and males were 3.37 mph and 3.20 mph, respectively, when the body mass were 333.79g and 291.36g. The month of inflection points for total length growth were 1.80 mph and 1.70 mph, respectively, when the total length were 18.52cm and 16.28cm. At 1.5-2.0 mph, SSD was not clearly demonstrated. At 3.0 mph, the body mass of the females was significantly higher than that of the males (P < 0.05), Serum E2, brain gh, and liver igf1 expression of the females was significantly higher than that of the males (P < 0.05); T content of the males was significantly higher than that of the females (P < 0.05). At 4.0months of age, the body mass of the females was highly significantly higher than that of the males (P < 0.01), Serum E2, brain gh, and liver igf1 expression of the females was highly significantly higher than that of the males (P < 0.05); T content of the males was significantly higher than that of the females (P < 0.05). With the continuous development of gonads, muscle and liver igfbp-5a and -5b expression generally tend to increase in females and males, while igfbp-5a showed a gradual increasing trend, and igfbp-5b expression showed a trend of decreasing and then increasing. Male igfbp-5a/b expression was significantly higher than female at the age of 3.0-4.0months (P < 0.05). This work verified that the females had faster growth rate since 3.0 mph compared to the males, which may be related to higher E2 levels in females leading to higher igf1 level, through inhibition of igfbp-5a/b expression.

A study on the mediating role of serum hormones in the effects of heavy metals on preeclampsia

Exposure to environmental heavy metals may pose a risk factor for developing preeclampsia (PE) modified through intervention. This case-control study aimed to investigate the association between serum heavy metal concentrations and PE in pregnant women and whether hormones served as mediating factors in the impact of heavy metals on PE. From October 2020 to 2022, 160 patients with PE and 160 pregnant women with normal deliveries were recruited at Dongguan Songshan Lake Central Hospital. Serum concentrations of manganese (Mn), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), cadmium (Cd), lead (Pb), β-human chorionic gonadotropin (β-hCG), progesterone (P), estradiol (E2), testosterone (T), cortisol (Cort), and cortisone (Cor) were measured. Logistic, restricted cubic splines, weighted quantile sum and multivariate linear regression models were employed to account for different aspects and explore the relationships among heavy metals, hormones, and PE. Mediation model analysis was performed to assess the role of hormones in mediation. The median concentrations of Mn, E2, and Cort were lower in the PE group than in the control group. The median concentrations of Cu, Zn, β-hCG, and T were higher in the PE than in the control. Mn, E2, and Cort showed negative associations with PE, while Cu, Zn, β-hCG, and T demonstrated positive associations, as determined through logistic regression. Mn, Cu, and Zn displayed linear dose-response relationships with PE. Zn and Cu had high weights in the positive association model of mixed heavy metal exposure with PE. The mediation analysis revealed that serum E2, P, T, Cort, and Cort/Cor might be potential mediators of the association between heavy metals (Mn, Cu, and Zn) and PE.

Impact of elevated serum estradiol levels before progesterone administration on pregnancy outcomes in frozen-thawed embryo transfer for hormone replacement therapy

ObjectiveThe objective of this retrospective cohort study is to investigate the impact of monitoring serum estradiol (E2) levels before progesterone administration within hormone replacement therapy (HRT) on pregnancy outcomes in women undergoing frozen-thawed embryo transfer (FET).MethodsAnalyzed HRT-FET cycles conducted at a reproductive center from 2017 to 2022. Serum E2 levels were measured prior to progesterone administration. Multivariate stratified and logistic regression analyses were performed on 26,194 patients grouped according to terciles of serum E2 levels before progesterone administration.ResultsThe clinical pregnancy rate (CPR) and live birth rate (LBR) exhibited a gradual decline with increasing serum E2 levels across the three E2 groups. Even after controlling for potential confounders, including female age, body mass index, infertility diagnosis, cycle category, number of embryos transferred, fertilization method, indication for infertility, and endometrial thickness, both CPR and LBR persistently showed a gradual decrease as serum E2 levels increased within the three E2 groups. The same results were obtained by multivariate logistic regression analysis.ConclusionsThis large retrospective study indicates that elevated serum E2 levels before progesterone administration during HRT-FET cycles are associated with reduced CPR and LBR post-embryo transfer. Therefore, it is advisable to monitor serum E2 levels and adjust treatment strategies accordingly to maximize patient outcomes.

Comparison of Day-Specific Serum LH, Estradiol, and Progesterone with MiraTM Monitor Urinary LH, Estrone-3-glucuronide, and Pregnanediol-3-glucuronide Levels in Ovulatory Cycles.

Background and Objectives: Fertility tracking apps and devices are now currently available, but urinary hormone levels lack accuracy and sensitivity in timing the start of the 6-day fertile window and the precise 24 h interval of transition from ovulation to the luteal phase. We hypothesized the serum hormones estradiol (E2) and progesterone (P) might be better biomarkers for these major ovulatory cycle events, using appropriate mathematical tools. Materials and Methods: Four women provided daily blood samples for serum E2, P, and LH (luteinizing hormone) levels throughout their entire ovulatory cycles, which were indexed to the first day of dominant follicle (DF) collapse (defined as Day 0) determined by transvaginal sonography; therefore, ovulation occurred in the 24 h interval of Day -1 (last day of maximum diameter DF) to Day 0. For comparison, a MiraTM fertility monitor was used to measure daily morning urinary LH (ULH), estrone-3-glucuronide (E3G), and pregnanediol-3-glucuronide (PDG) levels in three of these cycles. Results: There were more fluctuations in the MiraTM hormone levels compared to the serum levels. Previously described methods, the Fertility Indicator Equation (FIE) and Area Under the Curve (AUC) algorithm, were tested for identifying the start of the fertile window and the ovulation/luteal transition point using the day-specific hormone levels. The FIE with E2 levels predicted the start of the 6-day fertile window on Day -7 (two cycles) and Day -5 (two cycles), whereas no identifying signal was found with E3G. However, both pairs of (E2, P) and (E3G, PDG) levels with the AUC algorithm signaled the Day -1 to Day 0 ovulation/luteal transition interval in all cycles. Conclusions: serum E2 and (E2, P) were better biomarkers for signaling the start of the 6-day fertile window, but both MiraTM and serum hormone levels were successful in timing the [Day -1, Day 0] ovulatory/luteal transition interval. These results can presently be applied to urinary hormone monitors for fertility tracking and have implications for the direction of future fertility tracking technology.

Effect of electroacupuncture pre-conditioning on the expression rhythm of core clock gene Bmal1 in uterine tissue of controlled hyperstimulation rats.

To observe the effect of electroacupuncture (EA) pre-conditioning on the expression rhythm of clock gene Bmal1 in the uterine tissue of rats with controlled ovarian hyperstimulation(COH), so as to explore its mechanisms underlying improvement of the endometrial receptivity of ovarian superovulation during implantation. Seventy-two female SD rats with typical estrous cycles were randomly divided into normal control, model and EA pre-conditioning (pre-EA) groups, with 24 rats in each group. The COH model was established by giving the rats with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG) by intraperitoneal injection. The rats of the pre-EA group received EA stimulation (1 Hz/5 Hz, a tolerable strength) of "Guanyuan"(CV4) and "Sanyinjiao"(SP6) for 15 min each time, once daily (at 21：00 every day). After successive EA intervention during the first two estrous cycles, the modeling began in the third estrus cycle and the EA intervention was continued till the end of modeling, followed by raising the rats with superovulation induction and male rats undergoing vasoligation in one cage (1∶1). The rats during the estrum in the normal control group or those of the model group at the end of modeling were raised together with the male rats undergoing vasoligation in one cage. On the 5th day (04：00 AM) after raising in one cage, the rats in the three groups were sacrificed in six batches every 4 hours, with 4 rats in each group in each batch. The H.E. staining was used for revealing alterations of the endometrial thickness, number of glands and blood vessels and tissue histology, and ELISA employed to ascertain the contents of estradiol (E2) and progesterone (Pg) in serum. The expression rhythm of core clock gene Bmal1 ［In the present study, Zeitgeber time (ZT) is an artificially set laboratory time, i.e., ZT7 (07：00) is light on and ZT19 (19：00) is light off.］ and the expression of endometrial HoxA10 and leukemia inhibitory factor (LIF) mRNAs were detected by quantitative real-time PCR. The Western blot was employed to detect the expression levels of HoxA10 and LIF proteins. Findings of the clock gene Bmal1 level showed that the expression peak was at ZT12 and the valley value at ZT20 in the normal control group, and that of the peak value was at ZT20 and valley value at ZT12 in the model group, while in the pre-EA group, the peak value was at ZT8, and the valley value at ZT4. The difference of Bmal1 levels among the three groups was most significant at ZT12 (12：00), therefore, the tissue samples were taken at ZT12 in this study for comparison of the levels of different indexes among the 3 groups. Compared with the control group, the endometrial thickness, number of glands and blood vessels, HoxA10 and LIF mRNAs and proteins were significantly down-regulated (P<0.01, P<0.05), and contents of serum E2 and Pg were considerably up-regulated in the model group (P<0.01, P<0.05). Relevant to the model group, the pre-EA group had an apparent increase in the endometrial thickness, number of glands and blood vessels, and expression levels of HoxA10 and LIF mRNAs and proteins (P<0.05, P<0.01), and a marked decrease in the serum Pg (P<0.05). At the ZT12 (12：00 noon), compared with the normal control group, the mRNA level of Bmal1 was significantly decreased in the model group (P<0.01)；and compared with the model group, the level of Bmal1 mRNA was significantly increased in the pre-EA group (P<0.05). In addition, at the node of ZT16, the mRNA level of Bmal1 was significantly decreased in the model group in comparison with the normal control group (P<0.01). EA preconditioning can improve the endometrial receptivity during the implantation window period in rats with COH, which may be related to its functions in regulating the expression of clock gene Bmal1 in the uterine tissue and in correcting the disturbance of clock gene rhythm.

Effect of High Serum Estradiol Levels and Duration of Vaginal Estradiol Administration on Reproductive Outcomes in Frozen-Thawed Embryo Transfer Cycles

Background: The purpose of this study is to assess the effects of high serum estradiol (E2) levels on reproductive outcomes as well as to compare pregnancy outcomes of duration of vaginal E2 administration after the day of progesterone (P4) initiation in frozen-thawed embryo transfer (FET) hormonal replacement cycles. Methods: We performed a retrospective cohort study on 627 patients ≤40 years old that underwent a FET cycle. The patients were grouped based on the serum E2 level on the P4 initiation day as follows: E2 ≤600 pg/mL (n = 471); E2 ≥900 pg/mL (n = 151); the group of E2 600–900 pg/mL (n = 5) was not included in the analysis due to too small sample size. A total of 151 patients with E2 ≥900 pg/mL were grouped based on the duration of vaginal E2 administration after the day of P4 initiation: withdrawal before transfer (n = 53); constant administration (n = 98). Results: There was no significant difference in implantation rate, clinical pregnancy rate, spontaneous abortion rate, premature birth rate, live birth rate and multiple pregnancy rate between the two E2 level groups. The logistic regression analysis demonstrated that age was the only variable that significantly affected clinical pregnancy rate and live birth rate. There were no fetal malformations in either group. Baseline characteristics of patients with different duration of vaginal E2 administration were similar. The pregnancy outcomes showed no significant difference between the withdrawal before transfer group and the constant administration group. Conclusions: In FET cycles, high E2 level on the embryo transfer day did not reduce the clinical pregnancy and live birth rates. In addition, vaginal E2 could be stopped on the embryo transfer day since we found no benefit in continuing E2 until the pregnancy testing day.

P-701 Impact of vaginal microbiome on vaginal absorption of exogenous progesterone: a pilot study

Abstract Study question Does vaginal microbiome have an impact on vaginal absorption of exogenous progesterone (P intake) in artificial cycles? Summary answer Results suggest that the presence of lactobacillus in the vagina may positively impact the uptake of P. What is known already Low serum P levels on ET day decrease significantly live birth rates. About 20% of patients receiving micronized vaginal P show inadequate levels. It is of interest to find out which intrinsic factors might influence on the capacity of absorption (P intake) of vaginal P, and if they could be treated to prevent this situation. Vaginal microbiome hast been suggested as a possible factor, although this has not been addressed yet. The aim of this study was to explore if there is a correlation between vaginal microbiome and pH, and P intake. Study design, size, duration A prospective single-centre cohort pilot study including 92 ART patients was conducted from February 2022 to January 2023 in IVI-RMA Valencia. Embryo transfer was conducted in the context of a hormonal replacement therapy cycle with use of vaginal P (400mg/12h) for luteal phase support. “Progesterone intake” capacity was evaluated according to the vaginal microbiome status (lactobacillus dominant (LDM, &amp;gt;90%) or non-lactobacillus dominant (NLDM) and pH levels determinations (considered normal when moderately acidic ( &amp;lt; =4.8). Participants/materials, setting, methods Samples were taken twice: A) proliferative phase (day of initiation of exogenous P) and B) mid-luteal phase (day of ET, P + 5). Serum E2 and P levels as well as vaginal microbiome and pH were analysed. A validated qPCR methodology was used to accurately assess the presence and relative abundance of the four major lactobacillus species associated with eubiosis (crispatus, gasseri, iners, and jensenii) and the most common bacterial species implicated in dysbiosis. Main results and the role of chance Distribution of LDM and NLDM profiles were comparable between Samples A and B. No significant difference between any of the potential confounding factors (age, BMI,baseline P) between the lactobacillus CST groups (CST-I,-II,-III,-V &amp; lactobacillus) and the dysbiotic CST group (CST-IV) in Samples A and B were observed. Serum P levels on the ET day were significantly higher in LDM profiles both in Sample A (15.2 vs. 12.9ng/mL in NLDM;p=0.009) and B (15.0 vs. 12.8ng/mL;p=0.014). A positive significant correlation was found between serum P levels and Lactobacillus abundance in Samples A (r = 0.28; p = 0.008) and B (r = 0.30; p = 0.004). The relative abundance of Gardnerella vaginalis in Sample A was negatively related to serum P levels on the ET day (r=-0.24;p=0.02). Vaginal pH was significantly lower in LDM profiles both in Sample A (4.9 vs. 5.6 in NLDM; p = 0.002) and B (4.7 vs. 5.1;p&amp;lt;0.001). pH levels were negatively correlated with Lactobacillus abundance in Samples A (r=-0.34;p=0.0012) and B (r=-0.41;p&amp;lt;0.001). Additionally, serum P levels were negatively correlated with pH levels in Sample A (r=-0.14;p=0.191) and B (r=-0.27; p = 0.010). Indeed, when patients are grouped based on a cut-off of P = 8.8ng/mL, patients with a pH ≤ 4.8 have significantly higher P levels than patients with a pH &amp;gt; 4.8. Limitations, reasons for caution This is a pilot study, thus further trials with larger sample sizes should be performed in order to confirm these results. Additionally, vaginal microbiome alone does not explain the total variability in serum P levels measured in artificial cycles when using MVP. Wider implications of the findings We might have found one of the causes of the high heterogeneity on vaginal absorption of MVP in artificial cycles. This finding may help us with patient management when programming an ET in this type of cycles, improving personalised patient care and progressing on luteal phase support individualisation. Trial registration number 2110-VLC-095-EL

P-445 High serum estradiol on the day of frozen blastocyst transfer is associated with higher early miscarriage rates in hormonal replacement therapy cycles using transdermal estrogens

Abstract Study question Do serum estradiol(E2) levels on the day of frozen blastocyst transfer(FBT) affect the live birth rate (LBR) in hormonal replacement therapy(HRT) cycles using transdermal estrogens? Summary answer E2 levels ≥ 313 pg/mL on the day of FBT are associated with increased early miscarriage rates, but no significant impact on LBR. What is known already E2 plays a crucial role in endometrial receptivity during the periconceptional period and a massive role in placentation. However, the impact of serum E2 levels measured around the time of a FBT in HRT cycles outcomes remains controversial, with some studies showing a detrimental impact of high levels, and others underlining no difference. To this date, there is no study focusing on HRT cycles with transdermal estrogens only, though it is the safest way of estrogen administration regarding thrombo-embolic complications. Study design, size, duration Observational cohort study with 2364 patients undergoing HRT-FBT cycles at a university hospital, between January 2019 and December 2022. Women were only included once during the study period. Participants/materials, setting, methods Patients undergoing single autologous FBT under HRT using transdermal estrogens and vaginal micronized progesterone. The serum E2 level was measured in the morning of the FBT, in a single laboratory. The primary outcome was the LBR, secondary outcomes included clinical pregnancy rates, early miscarriage rates(EMR), and obstetrical outcomes. Patients were divided into three groups based on the 25th and the 75th percentiles of E2 levels and compared using univariate and multivariate logistic regression models. Main results and the role of chance A total of 2364 patients were divided into 3 groups: group A &amp;lt; 122 pg/mL (n = 590), group B 122-312 pg/mL (n = 1184), and group C ≥ 313 pg/mL (n = 590). Mean serum E2 level on the day of FBT was 247.8 pg/mL ± 209.01 pg/ml. The LBR was 32.0% (756/2364), and the early miscarriage rate was 26.3% (280/1075) in the overall population. Multivariate logistic regression showed a negative impact of high E2 levels (≥ 313 pg/mL) on EMR: an increase in the EMR was noted between group C and referent group B (30.3% versus 24.6%, OR 1.5 95%CI (1.06-2.17)). However, no significant impact of E2 levels was noted regarding the LBR (group C vs group B, adjusted OR 0.9 95%CI (0.69-1.10)), the clinical pregnancy rates, or the obstetrical outcomes (birthweight and term of delivery Limitations, reasons for caution The main limitation of our study is linked to its observational design. Extrapolation of our results to other laboratories, or other routes and/or doses of administering estrogens also needs to be validated. Wider implications of the findings Limiting high serum E2 levels on the day of FBT in HRT cycles may improve ART outcomes. Further studies are needed to evaluate if modifications of estrogen doses may improve pregnancy chances, in an approach to individualize the management of ART patients. Trial registration number not applicable

P-594 Retrospective comparative study of follitropin delta vs follitropin alfa in progestin-primed ovarian stimulation (PPOS) protocol in IVF

Abstract Study question Could the individualized fixed-dose algorithm with follitropin delta show a similar pregnancy rate to conventional dosing with follitropin alfa in PPOS protocol? Summary answer Individualized fixed-dose algorithm with follitropin delta showed a similar pregnancy rate to follitropin alfa In PPOS protocol. What is known already Follitropin delta (REKOVELLE, Ferring Pharmaceuticals, Switzerland) is the first human cell line driven recombinant follicle-stimulating hormone (FSH) with an individualized fixed-dose algorithm based on serum AMH level and body weight. Follitropin alfa is a Chinese hamster ovarian cell driven recombinant FSH and its dosing is adjusted according to follicular response during the stimulation period. There was no significant difference between follitropin delta and follitropin alfa in terms of pregnancy rate under GnRH antagonist protocol. Study design, size, duration A retrospective analysis of 355 patients aged under 40 years who underwent ovarian stimulation in PPOS protocol using follitropin delta or follitropin alfa and on whom frozen-thawed single blastocyst transfer was conducted between August 2021 and November 2023 was performed. Participants/materials, setting, methods 193 cycles of women who used follitropin delta were compared to 162 cycles of women who used follitropin alfa. The following clinical outcomes on each treatment group were analyzed: chemical pregnancy rate (serum β hCG), clinical pregnancy rate (gestational sac), ongoing pregnancy rate(gestational sac and fetal heartbeat), serum E2 level at trigger day, cycle proportion of ≥ 10 oocytes retrieval, blastocyst formation rate, cycle proportion of ≥ 5 blastocysts and cycle proportion of ≥ 2 good quality blastocysts. Main results and the role of chance Baseline demographics in the follitropin delta group and the follitropin alfa group were not significantly different: age, 33.9 ± 3.5 years vs 34.5 ± 3.2 years; body weight, 53.5 ± 8.3 kg vs 53.8 ± 5.9 kg; number of previous IVF cycles, 0.2 ± 0.6 vs 0.3 ± 0.7; serum AMH level, 5.5 ± 2.9 ng/mL vs 5.3 ± 3.8 ng/mL, respectively. The chemical pregnancy rate (69.9% vs. 64.8%, P = 0.308), clinical pregnancy rate (58.0% vs. 56.2%, P = 0.748) and ongoing pregnancy rate (51.8% vs. 46.3%, P = 0.338) were comparable between the follitropin delta group and the follitropin alfa group. The individualized follitropin delta treatment resulted in a smaller cycle proportion of ≥ 10 oocytes retrieval (77.7% vs. 90.8%, P &amp;lt; 0.001) with lower serum E2 level at trigger day (3273 ± 1577 pg/mL vs. 3927 ± 1732 pg/mL, p &amp;lt; 0.001). However, the blastocyst formation rate was significantly higher in the follitropin delta group (75.5% vs. 68.7%, p &amp;lt; 0.001). Moreover, the cycle proportion of ≥ 5 blastocysts (71.0% vs. 76.4%, P = 0.286) and the cycle proportion of ≥ 2 good quality blastocysts (51.8% vs. 60.3%, P = 0.115) were comparable between the groups. Limitations, reasons for caution This is a non-controlled, retrospective study. Wider implications of the findings Follitropin delta in its individualized fixed-dose regimen could have the potential to decrease the risk of OHSS by minimizing excessive ovarian response, whereas a similar pregnancy rate to conventional dosing regimen could be expected due to the number of good quality blastocysts secured. Trial registration number not applicable

Does Serum Estradiol Prior to Progesterone Administration in Programmed Frozen Embryo Transfer Cycle Impacts Live Birth Rate: A Prospective Cohort Study.

There is a paucity of data on the optimal threshold of oestradiol (E2) levels required for successful implantation as this is a valid question to be answered, considering that significant number of in vitro fertilisation cycles are deferred for FET cycles. The aim of this study is to investigate the serum E2 levels before starting exogenous progesterone in artificial autologous frozen thawed embryo transfers on pregnancy outcomes. This was a cohort study wherein clinical data of 213 artificial autologous programmed hormone-regulated frozen embryo transfer (HR-FET) cycles were collected. The HR-FET cycles were divided into three groups based on the E2 levels before starting progesterone: Group A: E2 ≤300 pg/mL, Group B: E2 301-500 pg/mL and Group C: E2 >500 pg/mL. After adequate endometrial stripe was appreciated by transvaginal ultrasonography, single blastocyst transfer was performed in all the cycles. The clinical pregnancy rate (CPR) in Groups A, B and C were 41.8%, 73.5% and 0.95%, respectively (P = 0.0001). The live birth rate (LBR) in Groups A, B and C was 18.1%, 62.2% and 0%, respectively (P = 0.0001). The miscarriage rate (MR) was significantly lower (15.3%) when E2 was between 301 and 500 pg/mL (P = 0.001). At both the extremes of serum E2 values, the rate of miscarriage increased. If serum E2 levels were ≤300 pg/mL or >500 pg/mL, the rate of miscarriage was 56.5% and 100%, respectively. Serum E2 levels in the range of 301-500 pg/mL before starting progesterone in programmed FET cycle are associated with an improved CPR and LBR and also decreased MR as compared to the extreme values of E2 ≤300 pg/mL or >500 pg/mL.

Discovery and evaluation of novel spiroheterocyclic protective agents via a SIRT1 upregulation mechanism in cisplatin-induced premature ovarian failure

Currently, no effective treatment exists for premature ovarian failure (POF). To obtain compounds with protective effects against POF, we aimed to design and synthesize a series of spiroheterocyclic protective agents with a focus on minimizing toxicity while enhancing their protective effect against cisplatin-induced POF. This was achieved through systematic modifications of Michael receptors and linkers within the molecular structure of 1,5-diphenylpenta-1,4-dien-3-one analogs. To assess the cytotoxicity and activity of these compounds, we constructed quantitative conformational relationship models using an artificial intelligence random forest algorithm, resulting in R2 values exceeding 0.87. Among these compounds, j2 exhibited optimal protective activity. It significantly increased the survival of cisplatin-injured ovarian granulosa KGN cells, improved post-injury cell morphology, reduced apoptosis, and enhanced cellular estradiol (E2) levels. Subsequent investigations revealed that j2 may exert its protective effect via a novel mechanism involving the activation of the SIRT1/AKT signal pathway. Furthermore, in cisplatin-injured POF in rats, j2 was effective in increasing body, ovarian, and uterine weights, elevating the number of follicles at all levels in the ovary, improving ovarian and uterine structures, and increasing serum E2 levels in rats with cisplatin-injured POF. In conclusion, this study introduces a promising compound j2 and a novel target SIRT1 with substantial protective activity against cisplatin-induced POF.

Metagenomic analysis revealed the association between gut microbiota and different ovary responses to controlled ovarian stimulation

The aim of this study was to assess the correlation between gut microbial taxonomy and various ovarian responses to controlled ovarian stimulation. A total of 22 IVF cycles with a follicle-to-oocyte index (FOI) < 0.5 and 25 IVF cycles with FOI ≥ 0.5 were included in this study. Baseline demographic characteristics were compared between the two groups. Metagenomic sequencing was performed to analyze fecal microbial community profiles. Mice were used to evaluate the effect of Bifidobacterium_longum on ovarian response to stimulation. Compared with FOI < 0.5 group, women in group with FOI ≥ 0.5 had significant more oocytes retrieved (p < 0.01). Prevotella_copri, Bateroides_vulgatus, Escherichia_coli and Bateroides_stercoris were more abundant in FOI < 0.5 group while Bifidobacterium_longum, Faecalibacterium_prausnitzii, Ruminococcus_gnavus and Bifidobacterium_pseudocatenula were more abundant in FOI ≥ 0.5 group. After adjusting for women’s age and BMI, Pearson correlation analysis indicated alteration of gut microbiome was related with serum E2, FSH, number of oocytes retrieved and clinical pregnancy rate. Animal study showed ovarian response will be improved after Bifidobacterium_longum applied. An increased abundance of Bacteroidetes and Prevotella copri, as well as a decreased abundance of Bifidobacterium longum, have been found to be associated with poor ovarian responsiveness. Changes in gut microbiomes have been observed to be correlated with certain clinical characteristics. The potential enhancement of ovarian response may be facilitated by the integration of Bifidobacterium longum.

Combined analysis of estradiol and β-hCG to predict the early pregnancy outcome of FET: a retrospective study

BackgroundThe accurate prediction of pregnancy outcomes in in vitro fertilization (IVF) cycles is crucial. While several studies have been conducted on the predictive power of serum estradiol (E2) and β-hCG concentrations post-embryo transfer (ET) for pregnancy outcomes, there is debate on the predictive value of E2. The objective of this study was to investigate the predictive efficacy of combining serum E2 and β-hCG levels on early reproductive outcomes 12 days after embryo transfer.MethodsA total of 1521 patients with β-hCG positive values on day 12 following frozen-thawed embryo transfer (FET) with natural endometrial preparation cycles (NCs) were gathered in affiliated Women’s Hospital of Jiangnan University. Using logistic regression, the relationship between pregnancy outcome and early serum E2 and β-hCG concentrations was examined. The receiver-operating characteristic (ROC) analysis was used to assess the predictive accuracy of the serum E2 and β-hCG concentrations.ResultsNotable distinctions were observed in the serum E2 and β-hCG levels on the twelfth day following FET with NCs between the groups classified as clinical pregnancy group (CP Group) and biochemical pregnancy group (BP Group). In addition, the cutoff values for E2 and β-hCG on day 12 following FET with NCs in cleavage embryo group (CE Group) were 129.25 pg/mL and 156.60 mIU/mL, respectively. The threshold values for E2 and β-hCG for the blastocyst group (B Group) were 174.45 pg/mL and 217.70 mIU/mL. Serum E2 day12 and β-hCG day12 were found to be substantially linked with clinical pregnancy by logistic regression analysis.ConclusionsSerum E2 and β-hCG concentrations were found to be significantly different between the CP Group and BP Group in infertility women underwent FET with NCs. Our retrospective cohort study’s findings suggest that the combination of early E2 and β-hCG levels on day 12 post-FET could be used as a predictive tool to evaluate the likelihood of both positive and negative pregnancy outcomes in FET with NCs.

Protective effects of probiotics against menopausal symptoms in ovariectomized mice

With growing interest in the role of the gut microbiome in the pathogenesis of menopausal disorders, probiotic supplementation is emerging as a novel therapeutic strategy for postmenopausal women. In this study, we investigated the protective effects of several probiotic strains against menopausal symptoms using an ovariectomized (OVX) mouse model fed a high-fat diet, in comparison with the effects of 17β-estradiol (E2) replacement. Among the eight tested probiotic strains, we selected four (Limosilactobacillus fermentum MG901, Lactiplantibacillus plantarum MG4604, Ligilactobacillus salivarius MG4679, and Lacticaseibacillus reuteri MG5463) that showed superior estrogen-like activity in an uterotrophic assay. We then examined their effects in OVX mice. The probiotic MG4679 was tested in combination with MG5463. During the 12-week experimental period, oral administration of the four probiotics reduced blood glucose and body temperature and ameliorated depression-like behavior in OVX mice. In the Barnes maze test, MG901 and MG4604 improved the spatial memory deficits observed in OVX mice. Micro-CT analysis of mouse femurs revealed that MG5463 prevented the OVX-induced reduction in cortical bone mineral density and bone volume fraction. Importantly, similar to E2 replacement, most of the test probiotics increased Akt phosphorylation in skeletal muscle, estrogen receptor-α (ER-α) protein expression in the liver, and ERK phosphorylation in the brain of OVX mice. However, unlike E2 treatment, all probiotics did not affect uterine weight, uterine ER-α expression, and serum E2 concentration. These results indicated that the probiotics used in this study exhibited protective effects against menopausal symptoms, partly through estrogen-mimetic action, without unfavorable uterine stimulation in OVX mice.

Clinical Use of Long-Term GnRH Agonist in Prepubertal Bitches: Effect on P4, E2 and Melatonin Serum Concentration before and During First Estrus

Abstract The aim of this study was to determine the length of the contraceptive period after the application of deslorelin implant (4.7 mg) in juvenile, prepubertal bitches and assess the effects on the hormonal status (P4, E2 and melatonin), as well as on the occurrence of normal physiological estrus. Twelve crossbred prepubertal female dogs (4 months of age) were divided into the two groups, the treated group (DESLO) and control (placebo) group (C). Hormone assays, vaginal microbiological and cytological smears and the measurements of electrical resistance of cervical mucus were performed once in 3 weeks until the first signs of proestrus, and every day during estrus. GnRH implant significantly postponed time of onset of estrus (633 ± 30,38 days vs. 143.80 ± 52,30 days, P&lt;0.001). In estrus, DESLO group showed statistically higher E2 (34.58 pg/mL vs. 20.59 pg/mL, P&lt;0.001) and melatonin serum concentration (45.86 pg/mL vs. 18.85 pg/mL, P&lt;0.001) compared to the control group. The average P4 serum concentration had no statistically significant differences between groups. In conclusion, deslorelin implants do not cause “flare up” effect in perpubertal bitches, however, increase of E2 and especially melatonin serum concentrations show the need to explore the consequences and mutual influence of melatonin and deslorelin implants in the future.