Research question:Embryo quality serves as a pivotal determinant in assisted reproductive technology (ART) outcomes, with high-grade embryos correlating with improved pregnancy/live birth rates but poor morphology elevating the risks of implantation failure and pregnancy loss. Despite its clinical significance, therapeutic options for enhancing embryo development remain limited, necessitating novel interventions to improve poor embryogenesis. Objective:This study aimed to evaluate the efficacy of DACHAO Reco18-a formulation containing natural extracts of clove, Sophora flower bud, and yam-in improving reproductive outcomes among IVF patients with a history of poor embryo quality. Design:This was a prospective self-controlled study, to assess the effects of DACHAO Reco18 containing natural extracts of clove, Sophora flower bud, and yam on reproductive outcomes in 68 IVF patients (aged 20-40 years) with a history of IVF/ICSI failure and documented poor embryo quality (≤20% high-grade embryos and no surplus embryos). All participants had previously undergone IVF/ICSI treatment at our center between January 2022 and December 2023. Over 6 weeks of oral treatment, outcomes were compared between pre- and post-intervention cycle. The primary outcome measure was embryo parameters, while secondary outcomes encompassed oocyte yield, fertilization efficiency, implantation rate and pregnancy rate. Results:DACHAO Reco18 intervention significantly improved the mean oocyte yield from 7.955 to 8.955 oocytes retrieved per cycle, fertilization outcomes with 2 pronucleus (PN) zygotes increasing from 3.731 to 5.463 embryos, and Day-3 embryo enhancement from 1.896 to 3.000 viable embryos. High-quality embryos surged by 118.52% from 0.567 to 1.239, with the quality rate improving from 30.729% to 44.107%. Additionally, when compared to the pre-treatment cycle, the post-treatment cycle showed statistically significant improvements in the implantation rate (0%-32.00%), biochemical pregnancy (0%-24.00%) and clinical pregnancy (0%-21.05%). Subgroup analyses revealed FSH reduction in hyper-gonadotropic patients (from 12.083 to 9.941 mIU/mL). Conclusion:This prospective self-controlled study suggests that a six-week DACHAO Reco18 intervention is associated with improved ovarian response and embryological outcomes in IVF patients with poor embryo quality, including reduced FSH in hyper-gonadotropic individuals. While promising, these preliminary findings warrant verification through randomized controlled trials to evaluate its potential as an adjuvant treatment in ART.

Non-invasive prenatal testing (NIPT) as a first-tier screening approach at population scale warrants close evaluation of its accessibility, affordability and extended application value. Here we present insights from a fully publicly funded NIPT programme for the pregnant population in Hebei province, China (July 2019–June 2022), with the latest NIPT coverage of 93.3%. Among 1,185,416 pregnant women, the incidences of fetal chromosome aneuploidies were 0.105% (trisomy 21), 0.026% (trisomy 18), 0.005% (trisomy 13), 0.056% (47,XXY), 0.043% (45,X), 0.039% (47,XYY) and 0.034% (47,XXX). Parental advanced age (≥35 years old) was related to a 4.31-fold risk for fetal trisomy 21 compared to parental non-advanced age. An elevated risk for small vulnerable newborns was observed in NIPT false-positive (relative risk: 1.49 (1.31, 1.68)) and NIPT high-risk without invasive prenatal diagnosis (relative risk: 1.85 (1.57, 2.18)) cases. NIPT reduced prenatal diagnosis by 97.4% compared to maternal serum screening, with better cost-effectiveness. Our findings indicate that first-tier NIPT for all pregnant women could be a feasible and cost-effective alternative to traditional maternal serum screening. NIPT positive results may be an early indicator of more adverse pregnancy outcomes beyond fetal aneuploidies. A retrospective analysis of a publicly funded NIPT programme in Hebei province, China, covering more than 1.18 million pregnancies, shows that first-tier NIPT achieved high coverage, reduced invasive testing by 97% and was more cost-effective than maternal serum screening, and the NIPT positive results were linked to higher risks of adverse pregnancy outcomes, suggesting potential early warning value.

OBJECTIVE:To test the hypothesis that women assigned to a natural ovulation regimen before frozen embryo transfer compared with a programmed regimen would have an increased chance of a healthy live birth and a reduced risk of pre-eclampsia or eclampsia. DESIGN:Multicentre, randomised, parallel group, assessor blinded clinical trial. SETTING:24 academic fertility centres in China. PARTICIPANTS:4376 ovulatory women (aged 20-40 years) planning to undergo a frozen single blastocyst transfer. INTERVENTIONS:Eligible participants were randomised (1:1) to receive a natural ovulation regimen or a programmed regimen of hormone replacement for endometrial preparation. Endometrial preparation and frozen embryo transfer timing were determined in the natural ovulation regimen group by monitoring natural follicle development and measuring serum levels of luteinising hormone, oestradiol, and progesterone. In the programmed regimen group, endometrial preparation was achieved by sequential administration of oestrogen and progesterone. MAIN OUTCOMES AND MEASURES:Primary outcomes were a healthy live birth and pre-eclampsia or eclampsia after a frozen embryo transfer. Secondary outcomes were cycle cancellation, biochemical pregnancy, clinical pregnancy, ongoing pregnancy, pregnancy loss, ectopic pregnancy, live birth, birth weight, and maternal, fetal, and neonatal complications. RESULTS:In the intention-to-treat analyses, 910 (41.6%) of 2185 patients in the natural ovulation regimen group and 890 (40.6%) of 2191 in the programmed regimen group achieved a healthy live birth (relative ratio 1.03 (95% confidence interval (CI) 0.96 to 1.10); P=0.49). The risk of pre-eclampsia was lower in the natural ovulation regimen group among patients who achieved clinical pregnancy than in the programmed regimen group (2.9% (38 of 1302) v 4.6% (61 of 1326); 0.63 (0.43 to 0.94); P=0.02). The incidences of early pregnancy loss (12.1% (158 of 1302) v 15.2% (201 of 1326); 0.80 (0.66 to 0.97)), placental accreta spectrum (1.8% (24 of 1302) v 3.6% (48 of 1326); 0.51 (0.31 to 0.83)), caesarean section (69.5% (776 of 1117) v 75.6% (831 of 1100); 0.92 (0.87 to 0.97)), and postpartum haemorrhage (2.0% (22 of 1117) v 6.1% (67 of 1100); 0.32 (0.20 to 0.52)) were lower in the natural ovulation regimen group. No differences between groups were observed for birth weight or neonatal complications. The rate of cycle cancellation was higher in the natural ovulation regimen (16.2% (354 of 2185) v 11.5% (251 of 2191), P<0.001). The prespecified per protocol and subgroup analyses yielded results consistent with the intention-to-treat analyses. CONCLUSIONS:In ovulatory women, a natural ovulation regimen for endometrial preparation was as effective as programmed regimen in terms of achieving a healthy live birth after frozen embryo transfer, but with a lower risk of maternal complications during pregnancy. TRIAL REGISTRATION:Chinese Clinical Trial Registry ChiCTR2200057990.

Objective:This study aims to assess the effects of intrauterine infusion of human umbilical cord blood mononuclear cells (HUCBMCs) on reproductive outcomes in patients with thin endometrium undergoing frozen embryo transfer (FET). Design:Prospective cohort study. Study Participants:This study enrolled 134 patients diagnosed with thin endometrium who underwent FET procedures between January 2022 and December 2023. The cohort was stratified into two groups: a treatment group receiving HUCBMCs infusion (n = 35) and a control group (n = 99). Methods:Participants were allocated into two groups: the HUCBMCs group and the control group, based on the administration of HUCBMCs intrauterine infusion. Reproductive outcomes were monitored for all participants. The primary outcome measure was live birth, while secondary outcomes encompassed endometrial characteristics (thickness, pattern, and endometrial-subendometrial (ES) blood flow distribution pattern), implantation rate, biochemical pregnancy, clinical pregnancy, ongoing pregnancy, miscarriage rate, and obstetric outcomes. Multivariable logistic regression analyses were employed to ascertain the independent effects of HUCBMCs. Results:No significant improvement in endometrial characteristics was noted following the infusion of HUCBMCs. Similarly, the live birth rate did not differ significantly between the groups, with the HUCBMCs group exhibiting a live birth rate of 27.6% (8 out of 29 patients), comparable to the control group 27.1% (19 out of 70 patients) (P = 0.964). Additionally, when compared to the control group, the HUCBMCs group showed no statistically significant differences in implantation rate, biochemical pregnancy, clinical pregnancy, and ongoing pregnancy, as determined by multivariate logistic regression analysis (OR 0.46, 95% CI 0.16-1.32; OR 0.74, 95% CI 0.24-2.26; OR 0.79, 95% CI 0.26-2.41; OR 0.96, 95% CI 0.13-7.22, respectively). Notably, the miscarriage rates were 38.5% in the HUCBMCs group and 36.7% in the control group. Moreover, blastocyst transfer was associated with a significantly higher likelihood of live birth compared to cleavage embryo transfer, as evidenced by both crude and multivariate logistic regression analyses (OR 6.52, 95% CI 2.22-19.20; OR 7.86, 95% CI 2.20-28.08). No adverse effects were observed in the HUCBMCs group. Conclusion:Our findings indicate that although HUCBMCs infusion was deemed safe, there was no statistically significant improvement in reproductive outcomes between the HUCBMCs group and the control group. The miscarriage rate remained high among patients with thin endometrium. However, blastocyst transfer may be associated with a higher live birth rate compared to cleavage embryo transfer.

Elevated maternal body mass index (BMI) increases miscarriage risk in assisted reproductive technology, but it is unclear whether this risk differs between singleton and twin pregnancies. Twin gestations impose greater metabolic and hemodynamic demands, which may amplify the adverse effects of elevated BMI. This study aimed to determine whether pregnancy plurality modifies the association between maternal BMI and miscarriage risk following frozen-thawed embryo transfer (FET). In this retrospective cohort study, 13,911 FET cycles performed at a tertiary reproductive medicine center between January 2019 and June 2024 were included. Maternal BMI was categorized as underweight (< 18.5 kg/m²), normal weight (18.5–24.9 kg/m²), overweight (25.0–29.9 kg/m²), and obese (≥ 30.0 kg/m²). The primary outcome was total miscarriage (pregnancy loss before 28 weeks), with secondary outcomes including early miscarriage (< 12 weeks), late miscarriage (12–28 weeks), clinical pregnancy and live birth. Generalized estimating equation (GEE) models were used to estimate adjusted odds ratios (aORs) with 95

Osteogenesis imperfecta (OI) is a genetically heterogeneous skeletal dysplasia mainly caused by mutations in the COL1A1 and COL1A2 genes, which encode type I collagen. Here, using whole-exome sequencing(WES), we identified a rare heterozygous missense mutation (c.1253G > C, p.Gly418Ala) in COL1A2 in a Chinese family with recurrent prenatal skeletal dysplasia. This variant is located within the highly conserved Gly-X-Y repeat of the collagen triple-helical domain and predicted to be deleterious by bioinformatics programs. However, according to the American College of Medical Genetics and Genomics (ACMG) guidelines, it was initially classified as a variant of uncertain significance (VUS). To validate its pathogenicity, we overexpressed the mutant in a zebrafish model. The results revealed that mutant mRNA induced significant phenotypic abnormalities in zebrafish larvae, including shortened body length, axial curvature, fin malformations, and reduced vertebral mineralization—recapitulating key features of human OI. Our findings confirm the pathogenic role of the p.Gly418Ala mutation and highlight the zebrafish model as a powerful tool for rapid functional validation of VUS, facilitating clinical variant interpretation and precision medicine.

Unexplained recurrent pregnancy loss (URPL), defined as two or more pregnancy losses without identifiable cause, affects approximately 2

STUDY QUESTION:In couples with non-severe male infertility, can sperm DNA fragmentation index (DFI) testing serve as a biomarker to identify couples who would benefit from ICSI over conventional IVF, based on cumulative live birth rate? SUMMARY ANSWER:Our secondary analysis of a randomized clinical trial (RCT) indicated that sperm DFI testing has limited value in identifying couples who would benefit from use of ICSI over IVF, based on cumulative live birth rate. WHAT IS KNOWN ALREADY:We recently demonstrated that in couples with non-severe male factor infertility, ICSI decreases cumulative live birth rates following the first transfer compared to IVF. However, it remains unclear whether sperm DFI testing can effectively guide treatment selection between ICSI and IVF. STUDY DESIGN SIZE DURATION:We used data of participants included in a randomized controlled trial comparing ICSI versus IVF in couples with non-severe male factor infertility. Participants had been recruited between April 2018 and November 2021 from seven centres in China, with follow-up outcomes collected as of 31 August 2023. PARTICIPANTS/MATERIALS SETTING METHODS:We included 953 of 2329 couples in whom the male partner had a baseline sperm DFI test from seven centres within the original RCT. The primary outcome was cumulative live birth, defined as a live birth following embryo transfers that occurred within 12 months after randomization within one oocyte retrieval cycle. The statistical analysis was performed based on an as-treated population, including 480 in the ICSI group and 473 in the IVF group. Multivariable fractional polynomial interaction analysis was performed to investigate non-linear interaction between DFI test and treatment effects of ICSI over IVF. MAIN RESULTS AND THE ROLE OF CHANCE:The median values of sperm DFI were 18.8% (interquartile range [IQR]: 12.2%∼26.3%) in the ICSI group, and 18.4% (IQR: 12.6%∼25.1%) in the IVF group. In each quantile of sperm DFI (Q1: DFI < 12.3%; Q2: 12.3≤DFI < 18.6%; Q3: 18.6≤DFI < 25.9%; Q4: DFI ≥ 25.9%), there were no significant differences of ICSI vs IVF on cumulative live birth (Q1: 36.7% vs 49.6%, adjusted odds ratio (aOR) = 0.61, 95% CI: 0.36 to 1.04; Q2: 51.8% vs 50.4%, aOR = 1.06, 95% CI: 0.60 to 1.84; Q3: 48.4% vs 56.3%, aOR = 0.78, 95% CI: 0.46 to 1.33; Q4: 45.2% vs 49.1%, aOR = 0.80, 95% CI: 0.47 to 1.37). Multivariable fractional polynomial interaction analysis showed no evidence of interaction between sperm DFI and the treatment effect of ICSI over IVF on cumulative live birth (P = 0.298). LIMITATIONS AND REASONS FOR CAUTION:First, sperm DFI testing was not routinely performed across all trial sites. Second, this study may be underpowered to detect differences in some outcomes, especially total fertilization failure, due to the very small number of events. WIDER IMPLICATIONS OF THE FINDINGS:Sperm DFI testing during the basal semen analysis has limited value in guiding the choice of fertilization methods for patients with non-severe male infertility. Based on the current evidence, sperm DFI testing should not be routinely recommended for this population. STUDY FUNDING/COMPETING INTERESTS:This study was funded by the National Key Research and Development Program (2022YFC2703102 to Y.W.), and the Peking University Third Hospital (BYSYDL2022001 to J.Q., BYSYDL2024003 to Y.W., and BYSYZD2019007 to Y.L.). B.W.M. reports consultancy, travel support, and research funding from Merck KGaA and consultancy for Organon and Norgine. TRIAL REGISTRATION NUMBER:The original trial was registered at Clinicaltrials.gov: NCT03298633.

PurposeThis study investigates the impact of the Chinese New Year (CNY) holiday on frozen embryo transfer (FET) outcomes in assisted reproductive technology (ART), particularly in vitro fertilization (IVF). Previous research has highlighted the negative effect of the CNY holiday on fresh embryo transfer outcomes, prompting an exploration of whether FET outcomes are similarly affected.MethodsA retrospective cohort study was conducted at the Second Hospital of Hebei Medical University, analyzing FET cycles performed from January 2012 to December 2022. A total of 4456 women were included, with 305 undergoing FET during the CNY holiday season and 4151 during non-holiday periods. The primary outcome measure was the live birth rate. Multivariate logistic regression and propensity score matching (PSM) were applied to assess the differences between the CNY and non-CNY (N-CNY) groups.ResultsMultivariate logistic analysis revealed no significant difference in live birth outcomes between the CNY and N-CNY groups (OR = 1.11, 95% CI 0.86 to 1.42). PSM analysis further confirmed that live birth rates were similar between the groups, with 39.0% in the N-CNY group and 44.2% in the CNY group (p = 0.208). These findings suggest that FET outcomes are not adversely affected by the CNY holiday.ConclusionFET outcomes during the CNY holiday season remain resilient, contrasting with findings from fresh embryo transfer studies. This may be attributed to the shorter duration and less intensive preparation of FET cycles, reducing the influence of sociocultural events and psychological stress. Further multicenter studies are needed to validate these findings and explore the impact of other significant sociocultural events on ART outcomes.

Oocyte/embryo defects can result in oocyte maturation arrest, fertilization failure, embryonic arrest, and infertility as well as recurrent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) failures. However, the genetic determinants of human oocyte/embryo defects remain largely unknown, and the overall genetic diagnostic yield for such defects has not been evaluated. Here, we performed exome sequencing in 3,627 patients with oocyte/embryo defects. We identified a total of 479 positive cases carrying variants in 37 known genes, indicating a diagnostic yield of 13.2%. Case-control association studies combined with gene set enrichment analysis identified 123 novel candidate genes responsible for oocyte/embryo defects. These results provide a comprehensive genetic landscape of human oocyte/embryo defects and highlight the clinical significance of genetic counseling in infertile patients with oocyte/embryo defects. Our study will lay the foundation for transforming the traditional clinical practice for failed IVF/ICSI attempts into genetic-based precision and personalized treatment for these patients.

Intimate partner violence (IPV) and infertility are significant global health and human rights concerns. Evidence has shown that infertile women face a higher risk of IPV, yet national-level data on its prevalence remain limited. This study aims to quantify the prevalence of IPV against infertile women in China and identify associated risk factors. A national cross-sectional study was conducted in China from October 2021 to August 2022, using multistage stratified cluster sampling. Infertile women were recruited from 30 reproductive centers across ten provinces. A self-administered questionnaire, including the Revised Conflict Tactics Scale (CTS-2) and the Scale of Economic Abuse-12 (SEA-12), measured IPV in the past 12 months and over a lifetime. A structural equation model (SEM) explored risk factors for past-12-month IPV victimization. A total of 12,392 infertile women were included. The weighted 12-month prevalence of IPV was 34.1

Few large-scale studies have systematically examined the effects of maternal exposure to fine particulate matter (PM2.5) on live birth in women undergoing in vitro fertilization (IVF). This study aimed to investigate the associations between ambient PM2.5 exposure and live birth in women treated with IVF, and determine critical periods, key failure events, and vulnerable populations affected by such exposure. We included 58,637 patients from six reproductive centers in China between 2016 and 2021. We defined six exposure windows and adopted logistic regression with random-effect models to investigate the associations between PM2.5 exposure and live birth. We further categorized live birth failure as implantation failure, biochemical pregnancy loss, and miscarriage, to determine at which stage PM2.5 exposure caused the live birth failure. Subgroup analyses were conducted by female age, ovarian response, embryo quality, and transplantation protocol. For each 10 ​μg/m3 increase in PM2.5 concentration during follicle growth phase, preantral-antral follicle phase, and antral-mature follicle phase, the odds ratios for live birth were 0.966 [95% confidence interval (CI): 0.938, 0.995], 0.967 (0.939, 0.996), and 0.978 (0.958, 0.998), respectively. PM2.5 during these stages was significantly associated only with an increased likelihood of implantation failure, highlighting adverse impact of ambient PM2.5 on early pregnancy outcome. In addition, we observed relatively stronger associations in women with poor ovarian response, compromised embryo quality, and those undergoing fresh or double embryo transfers. This large-scale population-based study demonstrated the detrimental effects of high PM2.5 exposure for IVF women, shedding light on clinical and public health practices.

Polycystic ovary syndrome (PCOS) patients typically undergo either an ovulation induction regimen or a programmed regimen for endometrial preparation before frozen embryo transfer (FET). However, the superiority of one approach over the other remains controversial. While previous studies suggest that the letrozole regimen may improve pregnancy outcomes, prospective studies are insufficient. Therefore, we designed a multi-center randomized controlled trial to compare the pregnancy outcomes between these two regimens in PCOS patients undergoing FET. This multicentre, randomised controlled, open-label trial included 155 PCOS patients from six hospitals in China between September 2022 and February 2024. Patients were randomised into either the letrozole ovulation regimen group (n = 81) or the programmed regimen group (n = 74) during FET cycles. Subgroup analysis was used among patients with single blastocyst transfer. The primary outcome was clinical pregnancy rate, with secondary outcomes including abortion rate, live birth rate, and other pregnancy and neonatal outcomes. Analysis of 155 FET women showed no significant difference in clinical pregnancy rates between the letrozole group (62.96 https://www.chictr.org.cn ). Registered on 31 July 2022.

Introduction: Gonadotropin‐releasing hormone antagonists (GnRH‐ant) are commonly used during controlled ovarian stimulation (COS) in the in vitro fertilization (IVF) process to prevent premature luteinization and to ensure follicles mature synchronously. The 2020 guideline from European Society of Human Reproduction and Embryology (ESHRE) recommends the use of GnRH‐ant for patients with varying levels of ovarian response. However, the question of how to manage the protocol pathway for these patients requires further investigation. Methods: The current clinical practice guideline (CPG) adheres to the World Health Organization's (WHO) recommended development process, and covers eight clinical questions, all focusing on the application of GnRH antagonists in COS. We conducted Cochrane‐standard systematic reviews, utilized GRADE for evidence certainty assessment, and employed GRADE Evidence‐to‐Decision Framework to derive recommendations. We pre‐defined clinically important outcomes, and the common critical outcomes shared by all clinical questions as follows: live birth rate, implantation rate, clinical pregnancy rate, and moderate to severe ovarian hyperstimulation syndrome (OHSS). We strictly followed the RIGHT guideline and AGREE‐II criteria throughout the CPG development. Recommendations: The guideline development group (GDG) agreed on 14 recommendations on the application of GnRH‐ant during COS. In summary, low certainty of evidence supported the benefits of using GnRH‐ant protocol in IVF patients with high ovarian response (HOR), following with very low certainty of evidence to use Oral Contraceptive Pill (OCP) or estrogen as pretreatment in normal ovarian response (NOR) patients, low certainty of evidence to use fixed GnRH‐ant protocol and very low certainty of evidence to use GnRH agonist (GnRH‐a) add on trigger. For patients using GnRH‐ant protocol, very low certainty of evidence in HOR patients and low certainty of evidence in NOR patients supported the “freeze‐all” strategy considering the potential risk associated with fresh embryo transfer. However, multiple fresh embryo transfer may still provide benefits in some cases.

The clinical implications of elevated luteinizing hormone (LH) levels in patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) fresh embryo transfer cycles remain unclear, particularly regarding oocyte maturation and pregnancy outcomes. We aimed to evaluate the impact of basal LH elevation on treatment outcomes and compare the efficacy of protocols involving gonadotropin-releasing hormone antagonist (GnRH-ant) and gonadotropin-releasing hormone agonist (GnRH-a) for ovarian stimulation in patients with PCOS undergoing in vivo IVF/ICSI. This single-center, retrospective cohort study included 1,269 patients with PCOS who underwent IVF/ICSI treatment following a flexible GnRH-ant or long GnRH-a protocol with a human chorionic gonadotropin trigger between January 2013 and December 2023. Primary outcomes included the number of retrieved oocytes, metaphase II (MII) oocytes, two pronuclear embryos, high-quality embryos, and the ovarian hyperstimulation syndrome (OHSS) rate. Secondary outcomes included clinical pregnancy, miscarriage, and live birth rates following fresh embryo transfer. Among the participants, 492 (38.77

Introduction Polycystic ovary syndrome (PCOS), recognised as the predominant aetiological factor in ovulatory dysfunction-related infertility, accounts for approximately 70% of anovulatory infertility cases. Patients with PCOS have significantly higher anti-Müllerian hormone (AMH) levels than their counterparts undergoing in vitro fertilisation (IVF) for non-PCOS indications (eg, male/tubal factors). Several studies have suggested that a high AMH level is associated with adverse pregnancy outcomes in IVF, particularly preterm delivery. However, most of these studies are retrospective studies, and their results are inconsistent. The majority of AMH measurements are conducted before pregnancy; however, AMH levels fluctuate dynamically during pregnancy. There is a pressing need for a well-structured prospective study to definitively establish whether high AMH levels during pregnancy are associated with IVF/intracytoplasmic sperm injection (ICSI) pregnancy outcomes in PCOS patients.Methods and analysis This prospective cohort study will be conducted at four reproductive medicine centres. The plan is to enrol 1,320 PCOS patients and 1320 non-PCOS women who undergo IVF/ICSI and achieve singleton clinical pregnancies. Serum samples will be collected at about 6 weeks of gestation to measure the serum AMH level. Follow-up visits will be conducted at 12, 28 and 37 weeks of gestation, delivery and 6 weeks after delivery to obtain information about pregnancy outcomes and complications. The primary outcome is preterm delivery.Ethics and dissemination The study was approved by the Medical Research Ethics Committee of Peking University Third Hospital (M2022618). Informed consent will be obtained from all patients. The results of this clinical study will be presented at scientific conferences and submitted to a peer-reviewed journal.Trial registration number ChiCTR2300068554.

Abstract Background The way of testicular tissue fixation directly affects the correlation and structural integrity between connective tissue and seminiferous tubules, which is essential for the study of male reproductive development. This study aimed to find the optimal fixative and fixation time to produce high-quality testicular histopathological sections, and provided a suitable foundation for in-depth study of male reproductive development with digital pathology technology. Methods Testes were removed from both sides of 25 male C57BL/6 mice. Samples were fixed in three different fixatives, 10% neutral buffered formalin (10% NBF), modified Davidson’s fluid (mDF), and Bouin’s Fluid (BF), for 8, 12, and 24 h, respectively. Hematoxylin and eosin (H&E) staining, periodic acid Schiff-hematoxylin (PAS-h) staining, and immunohistochemistry (IHC) were used to evaluate the testicle morphology, staging of mouse seminiferous tubules, and protein preservation. Aperio ScanScope CS2 panoramic scanning was used to perform quantitative analyses. Results H&E staining showed 10% NBF resulted in an approximately 15–17% reduction in the thickness of seminiferous epithelium. BF and mDF provided excellent results when staining acrosomes with PAS-h. IHC staining of synaptonemal complexes 3 (Sycp3) was superior in mDF compared to BF-fixed samples. Fixation in mDF and BF improved testis tissue morphology compared to 10% NBF. Conclusions Quantitative analysis showed that BF exhibited a very low IHC staining efficiency and revealed that mouse testes fixed for 12 h with mDF, exhibited morphological details, excellent efficiency of PAS-h staining for seminiferous tubule staging, and IHC results. In addition, the morphological damage of testis was prolonged with the duration of fixation time.

Objective In China, there is a unique type of second marriage (SM) family where the woman is remarried, and the man is experiencing his first marriage. Additionally, the woman is older than the man. Therefore, these families experience many challenges: psychological, emotional, and societal pressure. Such family is a typical sample for studying sociocultural and psychological stress influencing on outcome of assisted reproductive technology (ART). This study aimed to investigate the impact of social psychological stress on the live birth outcomes AR. Methods In this retrospective cohort, second marriage (SM) families who visited the Second Hospital of Hebei Medical University reproductive center between January 2012 to December 2022 were screened, and 561s marriage families (the SM group) with 5600 first marriage (FM) families (the FM group) were included undergoing their first ART cycles. The primary outcome of this study was the live birth rate (LBR). Results The live birth rate (LBR) of SM group (30.7%) is lower than that of the FM group (43.6%) (p < 0.01). After adjustment by logistic regression, the second marriage group (OR = 1.269, 95%CI 1.031-1.562, p = 0.025) were independent factors associated with the outcome of live birth. After propensity score matching (PSM), the live birth rate of SM group (28.7%) is lower than the FM group (35.9%) (0 = 0.011). Conclusion The SM family experience higher levels of social and psychological pressure, which lead to lower level of LBR than FM family.

Objective:To estimate the proportion of married women in China who intend to become pregnant given the country's pronatalist population policy and to investigate fecundity, with an emphasis on the influence of socioeconomic factors.Methods:A nationally representative survey of 12 815 married women aged 20 to 49 years (mean: 36.8 years) was conducted during 2019 and 2020. All completed questionnaires, 10 115 gave blood samples and 11 710 underwent pelvic ultrasound examination. Fertility intention was the desire or intent to become pregnant combined with engagement in unprotected sexual intercourse. We defined infertility as the failure to achieve pregnancy after 12 months or more of unprotected intercourse. We considered an anti-Müllerian hormone level < 1.1 ng/mL and an antral follicular count < 7 as indicating an abnormal ovarian reserve.Findings:Fertility intentions were reported by 11.9% of women overall but by only 6.1% of current mothers (weighted percentages). Fertility intention was significantly less likely among women in metropolises (odds ratio, OR: 0.38; 95% confidence interval, CI: 0.31-0.45) and those with a higher educational level (OR: 0.74; 95% CI: 0.62-0.88). Overall, 18.0% had experienced infertility at any time and almost 30% had an abnormal ovarian reserve on assessment. An abnormal ovarian reserve and infertility were less likely in women in metropolises (P < 0.05) but more likely in obese women (P < 0.05).Conclusion:The willingness of Chinese married women to give birth remained low, even with relaxation of the one-child policy.