Serum LH Research Articles

P-611 Progestin-Primed Ovarian Stimulation is a non-inferior alternative to the GnRH Antagonist Protocol in patients undergoing ovarian stimulation: a propensity score weighting analysis

Abstract Study question To evaluate the effectiveness of the progestin-primed ovarian stimulation (PPOS) protocol versus the gonadotropin-releasing hormone antagonist (GnRH-ant) protocol in ovarian stimulation. Summary answer The PPOS protocol with dydrogesterone provided similar embryo outcomes to the GnRH-ant protocol. The serum LH concentration during ovarian stimulation using PPOS was well-controlled. What is known already Since Yanping Kuang's proposal PPOS in 2015, several studies on the effectiveness of this treatment regimen have been published. Although it has been suggested that outcomes of the PPOS regimen were comparable to those of GnRH-ant regimen, the findings were limited due to differences in study design, patient characteristics, dosage and type of progestins. While medroxyprogesterone acetate is often used in PPOS studies, dydrogesterone has seen far less popularity. Additionally, dydrogesterone is a relatively weak LH inhibitor compared to MPA, leading to concern about the increased incidence of unexpected LH surge Study design, size, duration We conducted a retrospective study among women who underwent In Vitro Fertilization at the Assisted Reproduction Center, Tam Anh General Hospital, Vietnam from January 1st, 2022 to October 10th, 2022. Participants/materials, setting, methods We included 804 patients who underwent ovarian stimulation, among which 208 used the PPOS protocol with dydrogesterone (30 mg dydrogesteron per day from the beginning of stimulation until the trigger-day) and 598 women used the GnRH-ant protocol (cetrorelix 0.25 daily, from day 6 of ovarian stimulation until the trigger-day). In addition to unadjusted analysis, we used propensity score weighting to account for confounding that might possibly remain. Main results and the role of chance Baseline characteristics were comparable in both groups. In both unadjusted and adjusted analysis, the mean number of good cleavage embryos in PPOS (6.33) was non-inferior to GnRH-ant (6.44; unadjusted ratio of two means 1.02, 95%CI 0.92, 1.13). The trigger-day estradiol level in patients with PPOS was higher (4,420 vs 3,830 pmol/L in GnRH-ant) despite similar total follicle stimulating hormone dose and fewer days of ovarian stimulation. The overall number of oocytes retrieved, the number of MII oocytes, the number of cleavage embryos, the number of blastocysts, and the number of good blastocysts were comparable between the two protocols. None of the PPOS patients had an unexpected LH surge, and serum LH levels decreased slightly during ovarian stimulation. Limitations, reasons for caution Due to the nature of the retrospective study design, we did not record the progesterone level on the trigger-day in the PPOS regimen; therefore, identifying LH surge based on LH and progesterone levels is not feasible. Furthermore, we did not study the outcomes of embryo transfer. Wider implications of the findings These results supported the use of PPOS with dydrogesterone as a cheap and effective alternative for clinicians in clinical practice. In patients with an indication for freeze-all embryo, the PPOS protocol could be recommended. Trial registration number Not Applicable

O-296 Progesterone level is an alternative marker to detect ovulation time in natural cycle frozen-thawed embryo transfers

Abstract Study question Do serum progesterone or LH levels better predict the optimal timing for embryo transfer in true natural cycle frozen-thawed embryo transfers (NC-FET)? Summary answer Performing embryo transfer according to serum progesterone levels (PL) had similar ongoing pregnancy rates compared to embryo transfer according to serum LH levels. What is known already With efficient and safe embryo vitrification techniques, there is an increasing trend in frozen-thawed embryo cycles (FET) over fresh cycles. FET protocols are programmed cycle, NC-FET and modified NC-FET. NC-FET is increasing in popularity, since it is associated with favourable obstetric and perinatal outcomes. In NC-FETs, ovulation time is the critical parameter for synchronization of embryo and endometrium. In the literature, LH surge is the most commonly used test to define ovulation time. Study design, size, duration This prospective cohort study included 183 NC-FET cycles in an IVF clinic between March 2022 and November 2022. Main outcome measure is ongoing pregnancy. Participants/materials, setting, methods 183 subfertile women aged between 18-40, having regular menstruation with a cycle length between 24-38 days are included. Protocol for NC-FET included serial hormone measurements and ultrasound monitoring when the dominant follicle reached the diameter of 15-16 mm. Serum LH level ≥15 IU/l defined as ‘ovulation -1’ in Group 1 and blastocyst embryo transfer was performed after 6 days. Serum PL > 1 ug/ml was defined as the ‘ovulation day’ in Group 2. Main results and the role of chance: There were no significant differences in baseline characteristics including female age, female body mass index, infertility duration, infertility diagnosis and number of failed IVF cycles between two groups. Also, cycle characteristics regarding endometrial thickness, number of embryos transferred, embryo quality and PLs at transfer day were similar. The overall clinical pregnancy was 59.6% and ongoing pregnancy was 50.8%. The implantation rate was 61.7%. The clinical pregnancy and ongoing pregnancy of Group 1 and Group 2 were 49.5%, 50.5% and 47.3%, 52.7%, respectively (p > 0.05). PLs in order to detect ovulation time were calculated according to a mathematical modelling (PL(Ov )=a1 eb1 Ov )(Ov:ovulation day; a1,b1:regression coefficients; e:euler number≌2.718) described in our previous study. Embryo transfer timing was planned after 5,4 or 3 days based on PLs calculated by modelling. Limitations, reasons for caution Since this is the first study planning embryo transfer in NC-FET based on progesterone levels, sample size should be increased and also serial change of progesterone levels were unknown in women >40 years of age. Wider implications of the findings Progesterone has a particular increasing pattern that enables ovulation day prediction accurately and may be used instead of serum LH levels in timing of embryo transfer. Trial registration number NCT05690360

O-276 Fertility preservation and sperm quality in adolescent transgender patients prior to hormonal treatment

Abstract Study question This study investigated the take-up, hormonal profile and sperm quality in adolescents undergoing fertility preservation via different methods, prior to medical intervention for gender dysphoria. Summary answer Fertility preservation via surgical sperm retrieval (SSR) or masturbation, is possible in this group, however long-term data is required to check gamete health. What is known already As increasing numbers of adolescents with gender dysphoria (GD) start gonadotrophin-releasing hormone agonists (GnRHa) to delay puberty to minimise psychological distress. However, the uncertainty of the long-term effects of these medications highlights the importance of undergoing fertility preservation (FP) prior to starting any hormone treatment. Study design, size, duration Data for adolescents wanting to FP was prospectively maintained. A total of 122 patients were referred for FP, of which 23 declined (19%). Participants/materials, setting, methods Young people were counselled for FP and serum testosterone, FSH, and LH were recorded prior to providing semen samples. Semen samples were classified by concentration; normal >15mil/ml, oligospermia <15mil/ml, poor <1sperm/slide, azoospermia. Insufficient samples, or unwillingness to masturbate, meant SSR was offered in a stepwise manner using electroejaculation (EE), TESE and mTESE. Main results and the role of chance Most patients (n = 78, 64% - median age 16.7) were able to produce semen by masturbation, cryopreserving an average of 6.6 straws. Sperm concentration was normal in 44% of samples produced, 36% were oligozoospermic, 9% were poor sperm concentration, and 12% were azoospermic. Mean blood results showed: testosterone 12.32nmol/l, FSH 3.83 IU/L, LH 4.39 IU/L. 21 patients required SSR - median age of 14.9. EE was successful in 4 patients, 9 underwent TESE, and 8 underwent mTESE. 4 mTESE patients were azoospermic. Success rate was 77%, with an average of 5.5 vials stored. Semen parameters in this cohort were poor - however possibly adequate for ICSI. Hormone levels were similar to the masturbation cohort: mean testosterone 11.7nmol/l, FSH 3.4 IU/L, LH 3.5 IU/L. 3 patients used GnRHa pre-SSR. Following a washout period, 1 patient remained azoospermic despite a testosterone of 21 nmol/l. The other 2 patients had sperm found with abnormal morphology and motility. This is the largest UK cohort of transgender girls referred for FP. The results showed that semen parameters were abnormal in 67% of the masturbation samples produced, and within the SSR group, all patient’s testosterone levels were >8nmol/l, with an average of 5.5 vials saved. Limitations, reasons for caution The quantity of long-term data on whether adolescents end up using their gametes post FP is a factor to consider with this study, as more evidence is needed in order to display the actual long-term usage of gamete cryopreservation. Wider implications of the findings Masturbation is often possible in this group, but alternatively, SSR provides patients with a safe opportunity to potentially have their own biological children in the future. Trial registration number not applicable

O-292 Does ovulation trigger enhance live birth in a natural cycle for frozen embryo transfer: A randomized controlled trial

Abstract Study question Does ovulation trigger enhance pregnancy rate in a natural cycle (NC) for frozen embryo transfer (FET)? Summary answer Pregnancy rates after a FET are similar in a natural cycle with detection of a spontaneous LH surge or with a triggered ovulation. What is known already Several studies have explored whether one approach might appear to be more beneficial in terms of pregnancy or live birth rates, but with inconsistent results. Thus, the best protocol to identify the time of ovulation in NC and to plan the FET is still undefined. Study design, size, duration This was a bicentric, two-arm, randomized, controlled and open-label trial, including 108 patients for 18 months. In the spontaneous-NC group, the LH surge was defined as a rise of the serum LH level at least three times above the basal LH serum value with a serum progesterone level three days after the LH surge above 3 ng/mL. Participants/materials, setting, methods Patients included were aged 18- to 39-year-old, had regular cycles and were planned for an autologous frozen day-5 blastocyst transfer in a NC. They did not have endometriosis stage AFSr 3 nor 4 nor adenomyosis. Fifty-four patients per group were included. In the modified-NC group, a spontaneous LH surge was observed in 16 women before the hCG-trigger could be administrated: they were re-allocated to the spontaneous-NC for the per-protocol (PP) analysis. Main results and the role of chance Patients characteristics were comparable in both groups. Pregnancy rates were similar in the spontaneous-NC and the modified-NC group in the intention-to-treat (ITT) (41.3 versus 37.8% respectively, p = 0.85) and in the PP analysis (39.3% versus 40% respectively, p = 0.9). Ongoing pregnancy, miscarriage and live birth rates were also similar. Significantly more visits were needed in the spontaneous-NC (3.4 ± 0.81 versus 2.59 ± 1.0, p < 0.0001 in the ITT analysis and 3.18 ± 0.93 versus 2.63 ± 1.03, p = 0.0064 in the PP analysis). Limitations, reasons for caution Due to group redistribution (spontaneous LH surge before hCG trigger), the number of subjects in the modified-NC group is low for the PP analysis. This may reduce the power of the trial which may fail to detect a small difference. Wider implications of the findings The results reinforce previous data that showed similar efficacy of the s-NC and the m-NC. This allows the patient to choose from several options depending on her preference. Trial registration number NCT03428165

O-251 LIN28B gene expression is downregulated in spermatozoa of oligozoospermic men and associates with genetic variants previously linked to pubertal onset

Abstract Study question Is there an association between genetic polymorphisms in LIN28B with LIN28B expression and semen quality in spermatozoa from oligozoospermic men? Summary answer LIN28B is downregulated in spermatozoa of oligozoospermic men and explains 39% of sperm count variability. One SNP (rs314280) was associated with low LIN28B mRNA expression. What is known already Large-scale genomic studies have identified genetic variants in or near LIN28B to be robustly associated with pubertal traits such as age at menarche and age at voice break. The LIN28 family of genes are known to control cell division, growth and differentiation. The LIN28B orthologue is exclusively expressed at high levels in the testis and placenta; however, we have limited knowledge about the role of LIN28B in relationship to testicular function. Interestingly, other studies have showed that later onset of puberty is associated with poor semen quality, suggesting that pubertal development may influence adult testicular function. Study design, size, duration Twenty-nine oligozoospermic (cases) and fifty-four men with normal sperm concentration (controls) were recruited from patients seeking andrological work-up for male/couple infertility at a University Hospital. Subjects with abnormal karyotype, Yq-microdeletions, varicocele GIII-IV, hypogonadotropic hypogonadism, seminal infection, diabetes, BMI>35, excessive consumption of alcohol/drugs, testicular cancer, chemotherapy/radiotherapy were excluded. Semen samples were obtained for analysis (WHO, 2010) and sperm isolation by density gradient (PureSperm40®). Peripheral blood samples were analysed for reproductive hormones and used for DNA extraction. Participants/materials, setting, methods LIN28B and GAPDH-S mRNA abundance were measured by qRT-PCR (2-ΔCT method) in RNA isolated from purified sperm using SYBR® MasterMix (Takyon). Five SNPs in linkage disequilibrium (rs7759938, rs395962, rs314268, rs314277 and rs314280) were genotyped (TaqMan™ SNP Genotyping Assay). Statistical differences between groups were assessed by Mann-Whitney test and genetic association was analysed using “SNPassoc” package in R. Values are showed as median, Q1-Q3. Main results and the role of chance Cases and controls had similar age and BMI but as expected sperm concentration, morphology and vitality were differed between groups (9.9, 4.9-13.1 vs 82.8, 46.0-146.5 mill/ml; 1, 1-2 vs 3, 1-4% normal forms and 83%, 76-89 vs 86%, 82-90, respectively) (p < 0.05). No differences were observed in progressive motility, semen volume and days of abstinence. FSH and LH serum levels were higher in cases compared with controls (4.6, 2.6-7.8 vs 3.2, 2.5-4 mIU/ml and 4.2, 2.9-5.2 vs 3.1, 2.1-4.2 mIU/ml, respectively; p = 0.019 and p = 0.017). Relative expression of LIN28B was significantly diminished in cases compared with controls (0.0034, 0.0015-0.0103 vs 0.0446, 0.0122-0.1026; p < 0.001), while expression of the internal housekeeping control GAPDH-S was similar between groups. Linear regression model showed significant association of sperm concentration and total sperm count with LIN28B relative expression adjusted by age, abstinence time, % progressive motility and % normal morphology (beta=0.408, R2=0.241, p < 0.001 and beta=0.461, R2=0.393, p < 0.001). LIN28B rs314280 was associated with mRNA expression in the dominant model (p = 0.025) and two other SNPs (rs7759938, rs314268) showed the same trend (p = 0.09). Men with at least one minor allele of these 3 SNPs had a lower LIN28B mRNA abundance (p = 0.006; p = 0.07; p = 0.055). Limitations, reasons for caution Genotype frequency comparisons between cases and control were not conducted because of the small sample size. A larger cohort will be studied to address this assumption and association of genotypes with parameters of semen quality. Wider implications of the findings To our knowledge this study represents the first report on LIN28B expression and human testicular function. The significant direct association of LIN28B mRNA expression with sperm concentration suggests that LIN28B may be involved in spermatogonial proliferation. Trial registration number not applicable

Is blue light exposure a cause of precocious puberty in male rats?

Our study aimed to examine the effects of blue light exposure on prepubertal male rats' puberty and testis tissue. Eighteen 21-day-old male Sprague Dawley rats were divided into three groups consisting of six rats in each group: Control Group (CG), Blue Light-6 hours (BL-6), and Blue Light-12 hours (BL-12). CG rats were maintained with 12/12-hour light-dark cycles. The rats of BL-6 and BL-12 were exposed to blue light (450-470nm/irradiance level 0.03uW/cm2) for 6 hours and 12 hours, respectively. Rats were exposed to blue light until the first signs of puberty. The ELISA method was used to analyze the serum levels of FSH, LH, testosterone, DHEA-S, leptin, ghrelin, melatonin, glutathione, glutathione peroxidase, and malondialdehyde. Testes were dissected for histomorphological examination. The medians of the pubertal entry days of the CG, BL-6, and BL-12 were 38th, 30th, and 28th days, respectively. (p:0.001) The FSH, LH, and testosterone concentrations of all groups were similar. The FSH concentration increased as the LH concentration increased (r: 0.82 p: 0.001). The serum LH concentration increased as serum testosterone, and DHEAS decreased, respectively (r: -0.561, p: 0.01) (r:-0.55 p:0.01). Testicular lengths and weights of the BL groups were smaller compared to CG (p: 0.03),(p: 0.04). GPx was higher for BL-6 and BL-12 than the CG (p:0.021, p:0.024). Testis tissue was compatible with the pubertal period in all groups. As the blue light exposure time increased, spermatogenesis was suppressed, and capillary dilatation and edema in the testis tissue increased. Our study is the first to show the effects of blue light exposure on male rats' puberty process. And we showed that exposure to blue light and the duration of exposure lead to precocious puberty in male rats. The blue light exposure suppressed spermatogenesis, marked vasodilatation in the interstitial area of the testis, and disrupted the integrity of the basement membrane. These findings intensified with increasing exposure time.

Dose-Dependent Protective Effect of Hygrophila auriculata Seeds on Cyproterone Acetate-Induced Testicular Dysfunction.

Infertility affects 15% of global population. This study was designed to search out the most effective dose of chloroform fraction of hydro-ethanolic extract of Hygrophila auriculata seed to ameliorate cyproterone acetate (CPA)-treated male subfertility. The rats were made subfertile by CPA at the dose of 2.5 mg/100gm body weight for 45 days. The male subfertility represented by low sperm concentration, less motile, less viable, and less hypo osmotic tail swelled spermatozoa in CPA-treated group. Serum LH, FSH, and testosterone levels were significantly decreased in CPA-treated group in respect to control. Androgenic key enzyme Δ5,3β-HSD, 17β-HSD activities and gene expression pattern were also decreased significantly in respect to control. These antispermatogenic and antiandrogenic activities of CPA were significantly recovered after the treatment of Hygrophila auriculata at the dose of 2.5 mg, 5mg, and 10 mg/100gm body weight. CPA also generate oxidative free radical that indicated by altered catalase, superoxide dismutase, and peroxidase activities and protein expression pattern along with conjugated diene and thiobarbituric acid reactive substance levels in testis. Expression pattern of Bax and Bcl2 genes were deviated from control after CPA treatment. Significant diminution of body weight, organo-somatic indices, and SGOT, SGPT activities were observed in CPA-treated group. All these biomarkers significantly recovered towards control after the treatment of Hygrophila auriculata at different doses. More significant recovery was observed in 5 mg and 10 mg of chloroform fraction-treated group and 5 mg dose, i.e., the minimum therapeutic dose to recover the CPA-induced subfertility.

Stem cell therapy for premature ovarian insufficiency: a systematic review and meta-analysis of animal and clinical studies.

The aim of this systematic review and meta-analysis is to evaluate the efficacy of stem cell therapy in mouse models of POI and patients with POI. The PubMed, Web of Science, and Embase databases were searched from inception to February 2022 for relevant animal and clinical studies. The reference lists of the included reviews were manually searched to identify additional eligible studies. Data were independently extracted by two investigators, and disagreements were resolved by discussion. SYRCLE's risk of bias tool and the MINORS tool were used to assess the quality of animal and clinical studies by two independent investigators. All statistical analyses were conducted using Review Manager 5.3 software. A total of twenty animal studies and six clinical studies were included in this meta-analysis. In animal studies, the results showed that stem cells could improve hormone levels, follicle count, estrous cycle and pregnancy outcome. For hormone levels, stem cells increased serum E2 and AMH levels and decreased serum FSH and LH levels compared with the control group (serum E2 level: SMD: 5.05, 95% CI 4.21-5.90, P < 0.00001; serum AMH level: SMD: 4.42, 95% CI 3.06-5.79, P < 0.00001; serum FSH level: SMD: -3.79, 95% CI -4.87 to - 2.70, P < 0.00001; serum LH level: SMD: -1.31, 95% CI -1.65 to -0.96, P < 0.00001). All follicle counts, except for the antral follicle count, were significantly changed compared with the control group. (primordial follicle count: SMD: 4.61, 95% CI 3.65-5.56, P < 0.00001; primary follicle count: SMD: 3.35, 95% CI 1.08-5.63, P = 0.004; secondary follicle count: SMD: 3.23, 95% CI 1.92-4.55, P < 0.00001; total follicle count: SMD: 4.84, 95% CI 2.86-6.83, P < 0.00001; oocyte count: SMD: 7.56, 95% CI 5.92-9.20, P < 0.00001; atretic follicle count: SMD: -1.79, 95% CI -2.59 to -1.00, P < 0.00001). For the estrous cycle, stem cell therapy increased the number of estrous cycles (WMD: 2.72, 95% CI 2.07-3.37, P < 0.00001) and decreased the duration of the estrous cycle (WMD: -1.26, 95% CI -1.84 to -0.69, P < 0.0001) compared with the control group. For pregnancy outcomes, stem cell therapy increased the fertility rate (RR: 3.00, 95% CI 1.74-5.17, P < 0.0001) and litter size (WMD: 3.82, 95% CI 0.36-7.28, P = 0.03) compared with the control group. In animal studies, the asymmetric funnel plot of serum E2 and FSH levels indicated the possibility of publication bias. Unpublished and negative studies may be the source of publication bias. In clinical studies, the results showed that stem cell therapy could decrease serum FSH level (MD: -30.32, 95% CI -59.03 to -1.01, P = 0.04) and increase AFC (MD: 1.07, 95% CI 0.70-1.43, P < 0.00001), pregnancy rate (RD: 0.19, 95% CI 0.04-0.34, P = 0.01) and live birth rate (RD: 0.19, 95% CI 0.07-0.31, P = 0.001) in POI patients. In addition, there was no significant difference in menstrual function regained (RD: 0.22, 95% CI -0.03-0.46, P = 0.09), oocytes retrieved (MD: 1.00, 95% CI -0.64-2.64, P = 0.23) and embryos (MD: 0.80, 95% CI -0.15-1.76, P = 0.10) between different groups. This meta-analysis suggested that stem cell therapy might be effective in POI mouse models and patients and could be considered a potential treatment to restore fertility capability in POI patients.

Endocrine changes induced by GnRH immunization and re-stimulation of testicular function with a GnRH agonist in stallions

Resumption of testicular function after GnRH immunization varies among animals and some stallions regain fertility only after a prolonged time (Janett et al., 2009, Anim Reprod Sci 115:88e102). This study evaluated endocrine effects of GnRH immunization and subsequent re-stimulation with a GnRH agonist. We hypothesized that GnRH agonist treatment advances resumption of normal endocrine function in GnRH-vaccinated stallions. Shetland stallions were assigned to a GnRH vaccination group (Improvac; 400 µg/animal in weeks 0 and 4; n=6) or a saline control group (n=6). Blood was collected weekly for hormone analyses. Each vaccinated stallion was hemi-castrated together with an age-matched control when testosterone concentration was <0.3 ng/ml for two consecutive weeks (C1). Treatment of vaccinated stallions with the GnRH agonist buserelin (4 µg/day for 4 weeks and 8 µg/day for 6 weeks) started three weeks thereafter. Irrespective of treatment, the remaining testicle was removed when testosterone concentration increased to >0.5 ng/ml for two consecutive weeks in vaccinated stallions (C2; 9-10 weeks after start of buserelin treatment). Testicular tissues were stored at – 80°C and fixed in formalin for RT-qPCR and histology. Statistical comparisons were made by repeated measures ANOVA with group as between and time as within subject factor. Data on semen characteristics in these stallions have been published recently (Gautier et al., 2022, Theriogenology 194:27-34). Plasma testosteroneconcentration decreased after GnRH-vaccination and returned to control stallion concentrations during buserelin treatment (time p<0.001, time x group p<0.001, group p=0.001). Serum LH was undetectable after the second vaccination, increased during buserelin treatment but remained lower than in control stallions (time p=0.003, time x group p<0.001). Serum FSH concentration decreased after vaccination and increased with buserelin treatment in vaccinated but even more in control stallions after both hemi-castrations (time p<0.001, time x group p<0.001, group p=0.008). Relative mRNA abundance for LH receptor (LH-R) and aromatase (CYP19A1) were lower in testicular tissue of vaccinated than control stallions at C1 (p<0.001). At C2, LH-R mRNA abundance no longer differed between groups, whereas CYP19A1 mRNA remained lower in vaccinated than in control stallions (p<0.001). Relative FSH receptor (FSH-R) mRNA abundance was higher in vaccinated than in control stallions at C1 (p<0.001) and similar in both groups at C2. Immunolabelling for LH-R was lower in testes of GnRH-vaccinated compared to control stallions at C1 and C2 (p=0.007), whereas FSH-R protein expression at no time differed between groups. Aromatase immunostaining was reduced at C1 in vaccinated stallions and increased until C2 (time p=0.035, time x group p<0.0031, group p<0.001). In conclusion, endocrine testicular function can be advanced in GnRH-vaccinated stallions by daily low dose buserelin treatment.

Nomogram for Predicting Live Birth after the First Fresh Embryo Transfer in Patients with PCOS Undergoing IVF/ICSI Treatment with the GnRH-Ant Protocol.

Polycystic ovary syndrome (PCOS) is the leading cause of anovulatory infertility. A better understanding of factors associated with pregnancy outcomes and successful prediction of live birth after IVF/ICSI are important to guide clinical practice. This was a retrospective cohort study investigating live birth after the first fresh embryo transfer using the GnRH-ant protocol in patients with PCOS between 2017 and 2021 at the Reproductive Center of Peking University Third Hospital. A total of 1018 patients with PCOS were qualified for inclusion in this study. BMI, AMH level, initial FSH dosage, serum LH and progesterone levels on the hCG trigger day, and endometrial thickness were all independent predictors of live birth. However, age and infertility duration were not significant predictors. We developed a prediction model based on these variables. The predictive ability of the model was demonstrated well, with areas under the curve of 0.711 (95% CI, 0.672-0.751) and 0.713 (95% CI, 0.650-0.776) in the training cohort and validation cohort, respectively. Additionally, the calibration plot showed good agreement between the prediction and the observation (p = 0.270). The novel nomogram could be helpful for clinicians and patients in clinical decision-making and outcome evaluation.

Evaluation of The Effects of Carob (Ceratonia Siliqua L.) Fruits on the Puberty of Rats.

This study was planned to determine the effects of carob use on puberty because of the observation of early puberty or pubertal variants due to the long-term use of carob in our clinic. Forty-eight Wistar albino rats, on postnatal day 21, were assigned into two groups female (n=24) and male (n=24). Groups were divided into four groups Control, Carob-150, Carob-300, and Carob-600. Ceratonia Siliqua L. extract was given to rats in a 0.5% carboxymethylcellulose(CMC) solution. Carboxymethylcellulose (0.5 %) was given to the Control, Ceratonia Siliqua L. extract was given 150 mg/kg/day to the Carob-150, 300 mg/kg/day to the Carob-300, 600 mg/kg/day to the Carob-600 by oral gavage. The treatments were performed once daily until the first sign of puberty. Serum FSH, LH, Estradiol, Total Testosterone, Leptin, Glutathione, Glutathione peroxidase, and Malondialdehyde were studied with rat kits by the ELISA method. Testis, uterus and ovary tissue were examined histologically. The median time of preputial separation in male rats were 38th-31st-31st-31st days, Control, Carob-150, Carob-300, and Carob-600 (p=0.004). The median time of the vaginal opening day in female rats were 39th-31st-34th-31st days in the Control, Carob-150, Carob-300, and Carob-600 (p=0.059). FSH-LH-Testosterone(male)-Estradiol(female)-Leptin levels of groups are similar (p>0.05). Glutathione peroxidase levels were higher in the groups given C. Siliqua extract compared to the Control in both male (p=0.001) and female groups (p=0.008). Testicular and ovarian tissues were concordant with the pubertal period in all groups. As the dose of CS increased, it induced spermatogenesis and spermiogenesis, causing abnormal changes such as ondulation in the basement membrane, capillary dilatation, and increased congestion in male. Edema in the medulla gradually increased with increased dosage, and granulosa cell connections were separated in Carob-300 and Carob-600 in female. Our study demonstrated that C. siliqua caused early puberty and increased spermiogenesis and folliculogenesis. We discovered that antioxidant mechanisms might affect this. Further, it may cause tissue damage at high doses.

Is There Still a Place for Surgery in Patients with PCOS? A Review.

The surgical management of polycystic ovary syndrome (PCOS) represents an unclear option compared to medical therapy, and it is necessary to deepen the role of minimally invasive surgery, represented by laparoscopic ovarian drilling (LOD) and transvaginal hydrolaparoscopy (THL), for the treatment of PCOS in infertile women resistant to drug therapy and to establish its success in terms of ovulation and pregnancy rates. A search was performed in the main electronic databases (MEDLINE, EMBASE, Web of Science, PubMed, and the Cochrane Library) from 1994 to October 2022 in order to evaluate the role of surgery in patients with PCOS resistant to pharmacological treatment. Only original scientific articles in English were included. Seventeen studies were analyzed in this review. In all analyzed studies, more than 50% of the population underwent spontaneous ovulation after surgical treatment without great differences between the two surgical techniques (LOD and THL). More than 40% of patients delivered, with a higher rate after LOD, although eight ectopic pregnancies and sixty-three miscarriages were reported. A lower risk of adhesion formation after THL has been reported. No clear data regarding the effect of surgery on the regularization of the menstrual cycle has been described. A reduction in LH and AMH serum levels as well as the LH/FSH ratio compared to preoperative levels for both surgical techniques has been described. Despite the scarcity and heterogeneity of data, surgical therapy could be considered an effective and safe approach in the management of PCOS patients with resistance to pharmacological treatment who desire to become pregnant.

Subacute high-refined carbohydrate diet leads to abnormal reproductive control of the hypothalamic-pituitary axis in female rats

We previously reported that female rats placed on a diet containing refined carbohydrates (HCD) resulted in obesity and reproductive abnormalities, such as high serum LH concentration and abnormal ovarian function. However, the impacts at the hypothalamic-pituitary (HP) function, specifically regarding pathways linked to reproductive axis modulation are unknown. In this study, we assessed whether subacute feeding with HCD results in abnormal reproductive control in the HP axis. Female rats were fed with HCD for 15 days and reproductive HP axis morphophysiology was assessed. HCD reduced hypothalamic mRNA expression (Kiss1, Lepr, and Amhr2) and increased pituitary LHβ+ cells. These changes likely contribute to the increase in serum LH concentration observed in HCD. Blunted estrogen negative feedback was observed in HCD, with increased kisspeptin protein expression in the arcuate nucleus of the hypothalamus (ARH), lower LHβ+ cells and LH concentration in ovariectomized (OVX)+HCD rats. Thus, these data suggest that HCD feeding led to female abnormal reproductive control of HP axis.

Are systemic progesterone levels in true natural cycle euploid frozen embryo transfers with luteal phase support predictive for ongoing pregnancy rates?

Are serum progesterone (P4) levels on the embryo transfer (ET) day predictive of ongoing pregnancy (OP) following a single euploid blastocyst transfer in a natural cycle (NC) when luteal phase support is routinely given? In single euploid frozen ETs in NC, P4 levels on ET day are not predictive for OP, when luteal phase support (LPS) is routinely added after the ET. In an NC frozen embryo transfer (FET), P4 produced by the corpus luteum initiates secretory transformation of the endometrium and maintains pregnancy after implantation. There are ongoing controversies on the existence of a P4 cutoff level on the ET day, being predictive for the chance of OP as well as of the possible role of additional LPS after ET. Previous studies in NC FET cycles, evaluating and identifying P4 cutoff levels did not exclude embryo aneuploidy as a possible reason for failure. This retrospective study analyzed single, euploid FET in NC, conducted in a tertiary referral IVF centre between September 2019 and June 2022, for which measurement of P4 on the day of ET and the treatment outcomes were available. Patients were only included once into the analysis. Outcome was defined as OP (ongoing clinical pregnancy with heartbeat, >12 weeks) or no-OP (not pregnant, biochemical pregnancy, early miscarriage). Patients with an ovulatory cycle and a single euploid blastocyst in an NC FET cycle were included. Cycles were monitored by ultrasound and repeated measurement of serum LH, estradiol, and P4. LH surge was identified when a rise of 180% above the previous level occurred and P4 levels of ≥1.0 ng/ml were regarded as confirmation of ovulation. The ET was scheduled on the fifth day after P4 rise and vaginal micronized P4 was started on the day of ET after P4 measurement. Of 266 patients included, 159 (59.8%) patients had an OP. There was no significant difference between the OP- and no-OP-groups for age, BMI, and day of embryo biopsy/cryopreservation (Day 5 versus Day 6). Furthermore, P4 levels were not different between the groups of patients with OP (P4: 14.8 ng/ml (IQR: 12.0-18.5 ng/ml)) versus no-OP (P4: 16.0 ng/ml (IQR: 11.6-18.9 ng/ml)) (P = 0.483), and no differences between both groups, when P4 levels were stratified into categories of P4 levels of >5 to ≤10, >10 to ≤15, >15 to ≤20, and >20 ng/ml (P = 0.341). However, both groups were significantly different for the embryo quality (EQ), defined by inner cell mass/trophectoderm, as well as when stratified into three EQ groups (good, fair, and poor) (P = 0.001 and 0.002, respectively). Stratified EQ groups remained the only significant parameter influencing OP in the uni- and multivariate analyses (P = 0.002 and P = 0.004, respectively), including age, BMI, and P4 levels (each in categories) and embryo cryopreservation day. Receiver operator characteristic curve for the prediction of an OP revealed an AUC of 0.648 when age, BMI and EQ groups were included into the model. The inclusion of P4 measurement on ET day into the model did not add any benefit for OP prediction (AUC = 0.665). The retrospective design is a limitation. Monitoring serum P4 levels can be abandoned in NC FET cycles with routine LPS as they do not seem to be predictive of live birth. No external funding was used for this study. The authors state that they do not have any conflicts of interest. N/A.

The Role of G3BP1 Gene Mediates P38 MAPK/JNK Pathway in Testicular Spermatogenic Dysfunction Caused by Cyfluthrin.

In recent years, male infertility has received global attention and seriously affected the quality of human fertility, and pyrethroids (type II pyrethroids), as recognized environmental endocrine disruptors, may threaten male reproductive health. Therefore, in this study, we established an in vivo model for the development of testicular and germ cell toxicity induced by cyfluthrin and explored the role and mechanism of the G3BP1 gene-mediated P38 MAPK/JNK pathway in testicular and germ cell damage caused by cyfluthrin to find early and sensitive indicators and new therapeutic targets for the development of testicular damage. Firstly, 40 male Wistar rats (about 260 g) were divided into a control group (corn oil), low dose group (6.25 mg/kg), middle dose group (12.5 mg/kg) and high dose group (25 mg/kg). The rats were anesthetized and executed after 28 days of poisoning on alternate days. Then, HE staining, transmission electron microscopy, ELISA, q-PCR, Western blot, immunohistochemistry, double-immunofluorescence and TUNEL were used to observe the pathology, androgen levels, oxidative damage and altered expression of the key factors of the G3BP1 and MAPK pathways in rat testes. The results showed that, compared with the control group, the testicular tissue and spermatocytes were superficially damaged with an increasing dose of cyfluthrin; furthermore, it could interfere with the normal secretion of the hypothalamic-pituitary-gonadal axis (serum GnRH, FSH, T and LH levels) and cause hypergonadal dysfunction. A dose-dependent increase in MDA and a dose-dependent decrease in T-AOC indicated that the oxidative-antioxidative homeostatic balance was disrupted. The Western blot and qPCR analysis revealed that G3BP1, p-JNK1/2/3, P38 MAPK, p-ERK, COX1 and COX4 proteins and mRNA expression were decreased, and p-JNK1/2/3, p-P38MAPK, caspase 3/8/9 proteins and mRNA expression were significantly increased. The double-immunofluorescence and immunohistochemistry results showed that the protein expression of G3BP1 decreased with an increasing dose of staining, while the expression of JNK1/2/3 and P38 MAPK were increased significantly. The positive expressions of G3BP1 were mainly located in the testicular germinal epithelium and germ cell layer, and the positive expressions of JNK1/2/3 were mainly located in the testicular germinal epithelium and sperm cells, while the positive expressions of P38 MAPK were located in all levels of the germ cells and spermatozoa. Our results demonstrated that exposure to cyfluthrin caused testicular and spermatocyte damage in rats, which could cause pathomorphology, altered androgen levels and a decreased antioxidant capacity. When the intracellular antioxidant capacity was impaired, G3BP1 expression and activity were inhibited, causing activation of the P38 MAPK/JNK pathway and activation of the intracellular apoptotic pathway, which, in turn, led to germ cell apoptosis.

Role of LIF genotypes on basal and day of triggering hormonal profile in women undergoing IUI

Female infertility is caused by ovulation issues, tubal disorders, uterine abnormalities, endometriosis, and peritoneal factors. For years, intrauterine insemination has been used to treat infertility alone or with ovarian stimulation. Obtaining adhesion ligands and losing inhibitory components are necessary for pregnancy. LIF promotes and differentiates proliferation and differentiation in embryonic, hematopoietic, and endothelial cells. LIF also controls prostaglandins (PGs), which mediate implantation and decidualization. The measuring hormone profile on cycle day 2 before control ovarian stimulation including FSH, LH, E2, Prolactin, TSH, and AMH. Serum LH, E2. The progesterone level was estimated on the day of triggering. The effect of LIF gene genotypes on hormone level. The hormone levels of 100 intrauterine inseminated women were measured on the second day of the cycle and the day of ovulation. DNA extraction for LIF gene variation detection utilizing sequencing and HRM. The current study found no significant differences between pregnant and non-pregnant groups for hormone levels, genotype, and genetic effect on female hormone levels with (P ≥0.05). The LIF gene did not affect hormone levels in infertile females may be due to the tiny samples or the complex signaling chain that happens during implantation

Insulin-Like Factor 3 (INSL3) in Serum and Follicular Fluid: Realtion With Marker Of Ovarian Reserve

Granulosa cells produce most ovarian function indicators in infertile women like estradiol, AMH, and inhibin B. Female peptide hormone insulinlike factor 3 is produced by theca cell. Signaling of insulin-like factor 3 from theca cell stimulates antral follicle development and theca androgen production. To investigate how INSL3 levels may be a theca cell marker of ovarian reserve and the association with current markers of ovarian reserve, including AFC, FSH, E2, and AMH, in infertile women undertaking assisted reproduction. In this cross-sectional study, 75 Iraqi infertile couples were chosen at Al-Nahrain University's High Institute for Infertility Diagnosis and Assisted Reproductive Technologies from October 2022 to April 2023 for the ICSI cycle. Based on the levels of INSL3 in the serum on the day 2 of the menstrual cycle and on the day of retrieving oocyte in the follicular fluid. A comparison was made with current ovarian reserve markers (AFC, FSH, E2, and AMH) .From 75 infertile women undergoing intracytoplasmic sperm injection (ICSI), 21 of them achieved pregnancy. Positive Significant relations were found between serum INSL3 levels and anti-mullerian hormone (AMH and antral follicle count. there were no significant relationships between follicular fluids INSL3 with serum LH, FSH and E2 . The study provides compelling evidence for the significant role of INSL3 in female fertility and its association with the success of intracytoplasmic sperm injection (ICSI). Positive correlations between serum INSL3 levels and antimullerian hormone (AMH) and Antral follicle count AFC indicate its potential involvement in ovarian reserve and hormonal regulation

Diagnostic Performance of Anti-Müllerian Hormone, Luteinizing Hormone to Follicle-Stimulating Hormone Ratio, Testosterone, and Prolactin to Predict Polycystic Ovary Syndrome Among Sudanese Women.

Polycystic ovary syndrome (PCOS) is one of the most important contributing factors to infertility. The diagnosis of PCOS is not an easy procedure, as the signs and symptoms are heterogeneous and of undefined etiology. There are only a few published studies that address the diagnostic performance of anti-Müllerian hormone in diagnosis of PCOS in sub-Saharan Africa including Sudan. This study aims to assess anti-Müllerian hormone (AMH), luteinizing to follicle-stimulating hormone ratio (LH: FSH), total testosterone (TT), and prolactin (PRL) levels among PCOS. In addition, we determine if AMH can be used as a predictor of PCOS among Sudanese women. There were 600 women enrolled in this observational cross-sectional study, 300 of whom had PCOS, and 300 of whom healthy women; PCOS was diagnosed using the Rotterdam criteria. On days 2-4 of the menstrual cycle, serum LH, FSH, AMH, TT, and PRL levels were measured for all participants. Diagnostic performance of these parameters for PCOS was determined by receiver operating characteristic (ROC) curve. Significantly higher means among PCOS regarding their BMI, AMH; LH: FSH ratio; TT; PRL, whereas significantly inverse in FSH compared with normal ovulatory women. On ROC analysis, AMH had the largest operating characteristic curve at cut-off >3.95 ng/mL; AUC = 0.999 with Youden's index 0.99%, followed by LH: FSH ratio at cut-off 0.749; AUC=0.932; Youden's index 0.813%, TT cut-off 0.82 mIU/L, AUC=0.852 with Youden's index 0.58, while PRL showed the lowest AUC=0.627 with cut-off 15.3 ng/mL, Youden's index was 0.18%, P. value<0.001. Sudanese women with PCOS had higher serum AMH level, LH:FSH ratio, and TT level. Moreover, AMH level has better discriminative power and good diagnostic potency for the diagnosis of PCOS among Sudanese.

Pre- and postnatal developmental exposure to the polychlorinated biphenyl mixture aroclor 1221 alters female rat pituitary gonadotropins and estrogen receptor alpha levels

Polychlorinated-biphenyls (PCBs) are industrial compounds, which were widely used in manufacturing of electrical parts and transformers. Despite being banned in 1979 due to human health concerns, they persist in the environment. In humans and experimental model systems, PCBs elicit toxicity in part by acting as endocrine-disrupting chemicals (EDCs). Aroclor 1221 (A1221) is a weakly estrogenic PCB mixture known to alter reproductive function in rodents. EDCs can impact hormone signaling at any level of the hypothalamic-pituitary-gonadal (HPG) axis, and we investigated the effects of A1221 exposure during the prenatal and postnatal developmental periods on pituitary hormone and steroid receptor expression in female rats. Examining offspring at 3 ages, postnatal day 8 (P8), P32 and P60, we found that prenatal exposure to A1221 increased P8 neonate pituitary luteinizing hormone beta (Lhb) mRNA and LHβ gonadotrope cell number while decreasing LH serum hormone concentration. No changes in pituitary hormone or hormone receptor gene expression were observed peri-puberty at P32. In reproductively mature rats at P60, we found pituitary follicle stimulating hormone beta (Fshb) mRNA levels increased by prenatal A1221 exposure with no corresponding alterations in FSH hormone or FSHβ expressing cell number. Estrogen receptor alpha (ERα) mRNA and protein levels were also increased at P60, but only following postnatal A1221 dosing. Together, these data illustrate that exposure to the PCB A1221, during critical developmental windows, alters pituitary gonadotropin hormone subunits and ERα levels in offspring at different phases of maturation, potentially impacting reproductive function in concert with other components of the HPG axis.

Effect of Ramadan Fasting on Sex Hormones in Infertile Male

Ramadan fasting is one of the 5 pillars of Islam and one of the most significant Ibadat (worships) of Islam. Throughout the world, millions of Muslims fast during Ramadan to fulfill this religious obligation. Aims: The aim of the study is to determine changes in the serum concentrations of gonadotropins, testosterone and prolactin hormones in infertile males after fasting of Ramadan and whether fasting improves fertility in the male. Participants and Methods: This is a case series study was conducted on 54 infertile male subjects aged (25-50 years) who fasted during Ramadan. Two blood samples were taken from each patient, one before and the other after fasting of Ramadan. Serum FSH, LH, prolactin and testosterone measurement was done by using Enzyme Linked Immune Sorbent Assay (ELISA). Results: There was no significant difference in serum FSH, LH, prolactin hormones when we compared them before and after Ramadan fasting. Concerning serum testosterone hormone, there was a decrease in its level after fasting but not reach significance when compared with its level before fasting. Conclusion: Serum levels of FSH, LH, prolactin and testosterone were unchanged after fasting of Ramadan.