BACKGROUND:Endometrial receptivity is crucial to a successful pregnancy. Although molecular markers of endometrial receptivity can be obtained through endometrium biopsy, the prediction method is invasive and delayed. Hence, it is important and necessary to establish a less invasive and more reliable method for the determination of endometrial receptivity to increase the efficiency and effectiveness of assisted reproductive technology (ART) treatments. METHODS:This study was performed in the Centre for Assisted Reproduction of Shanghai First Maternity and Infant Hospital between June 2021 and March 2022. Samples left on the transfer catheter after cleavage stage embryos transfer (day 3, D3, presumable pre-receptivity, n = 96) and blastocysts transfer (day 5, D5, presumable receptivity, n = 127) in 223 ART patients who achieved successful clinical pregnancies subsequently were collected. The collected samples were further divided into a training set (n = 111 samples) and a test set (n = 112 samples) for endometrial receptivity prediction model training and validation, respectively. RESULTS:All the collected samples contained sufficient total RNA for further analyses. Ultimately, 57 signature genes specifically expressed in the epithelial cells and the macrophages were identified. The endometrial receptivity predictability of our model in the training set exhibited an area under the curve (AUC) of 1.00, indicating complete stratification of pre-receptive and receptive samples. Likewise, in the test set, our model achieved an AUC of 0.91 with accuracy of 0.83, specificity of 0.86 and sensitivity of 0.79. CONCLUSIONS:Our study demonstrates a non-invasive method of endometrial sample collection for accurate determination of endometrial receptivity.

BACKGROUND:Oocytes from older females have a diminished ability to repair sperm DNA damage compared with those from younger females. Previous research has indicated that there is no significant correlation between sperm DNA fragmentation index (DFI) and the blastocyst euploidy rate in cycles utilizing oocytes donated by young individuals. However, it is still unclear whether a high DFI impacts the euploidy rate of blastocysts derived from oocytes obtained from women of advanced reproductive age. OBJECTIVE:The aim of this study was to investigate the potential association between sperm DFI and the euploidy rate of viable blastocysts in women of advanced age undergoing preimplantation genetic testing for aneuploidy (PGT-A). MATERIALS AND METHODS:A total of 667 blastocysts from 492 couples, all with maternal ages of 38 years or older, who underwent intracytoplasmic sperm injection (ICSI) combined with PGT-A were included in this study. The sperm DFI values were measured using the Sperm Chromatin Structure Assay (SCSA), and the couples were divided into three groups based on sperm DFI values: low DFI (DFI < 15%), moderate DFI (15% ≤ DFI ≤ 30%), and high DFI (DFI > 30%). RESULTS:No statistically significant differences were found in the rates of normal fertilization among the low, moderate, and high DFI groups (73.3%, 75.8%, and 75.4%, respectively; p > 0.05). Similarly, the rates of high-quality embryos were comparable among the groups (47.2%, 45.5%, and 45.7%, respectively; p > 0.05). The blastocyst formation rates also exhibited no significant differences among the groups (49.9%, 48.0%, and 49.3%, respectively; p > 0.05). Additionally, the aneuploidy rates of viable blastocysts were comparable across the groups (55.1%, 59.1%, and 60.6%, respectively; p > 0.05). Both the categorical analysis based on clinical DFI thresholds (<15%, 15%-30%, >30%) and the multiple linear regression treating DFI as a continuous variable (adjusted for female age, female body mass index [BMI], duration of infertility, number of miscarriages, and male age) revealed no statistically significant association between DFI and blastocyst euploidy rates (p = 0.733 for the categorical analysis; B = -0.003, standard error [SE] = 0.002; p = 0.136 for the continuous model). Furthermore, clinical pregnancy outcomes and neonatal results following the transfer of euploid blastocysts were comparable among the groups. DISCUSSION AND CONCLUSION:This study's findings suggest that elevated sperm DFI, as measured by the SCSA, does not significantly influence the euploidy rate of viable blastocysts in couples with advanced maternal age, whereas maternal age remains the predominant factor influencing embryo euploidy.

Research questionDoes the presence of smooth endoplasmic reticulum aggregates (SERa) in oocytes adversely impact the euploidy rate of subsequent blastocysts?DesignWe performed a retrospective cohort study with 671 young patients (< 38 years) undergoing their first preimplantation genetic testing for aneuploidy (PGT-A) between January 2019 and October 2022 at a reproductive medical center of university affiliated teaching hospitals in China. Cycles were categorized as either SERa(+) cycles (containing at least one SERa(+) oocyte) or SERa(-) cycles (all oocytes without SERa). In SERa(+) cycles, oocytes were further subdivided into the SERa(+) oocyte group and the sibling SERa(-) oocyte group, comprising oocytes with normal morphology.ResultsNo significant differences were observed in the normal fertilization rate (72.9% vs. 75.4% vs. 72.6%, P=0.343), and cleavage rate (96.8% vs. 97.1% vs. 96.4%, P=0.839) among the SERa(-) cycle group, the SERa(-) oocyte group, and the SERa(+) oocyte group. Additionally, there were no statistically significant differences in the rates of good quality embryos (44.7% vs. 48.8% vs. 46.2%, P=0.177) or blastocyst formation (60.1% vs. 60.9% vs. 60.5%, P=0.893) among the groups. However, the euploidy rate of blastocysts derived from SERa(+) oocytes was significantly lower compared to those from SERa(-) oocytes in SERa(+) cycles and normal oocytes in SERa(-) cycles (39.3% vs. 51.2% vs. 54.5%, P=0.005). Despite this, there were no significant differences in pregnancy and neonatal outcomes after euploid embryo transfer among the three groups.ConclusionsBlastocysts derived from SERa(+) oocytes have a lower euploidy rate than those derived from SERa(-) oocytes. Nevertheless, comparable reproductive outcomes were achieved following euploid embryo transfer from both SERa(+) and SERa(-) oocytes.

Background: Polycystic ovary syndrome (PCOS) is characterized by excess androgens, ovulatory dysfunction, and polycystic ovaries. The mechanisms underlying ovulatory and metabolic disorders in PCOS remain elusive, hampering therapeutic development. Enhanced metabolic health correlates with increased microbiota gene content and microbial diversity. We aimed to explore the impact of gut microbiota and serum steroids on PCOS regulation associated with androgen excess.Methods: The fecal samples of patients with hyperandrogenic PCOS (n = 14) and control group with PCOS (n = 14) were analyzed by 16S rRNA gene sequencing. The peripheral venous blood of all subjects was collected to detect serum hormones. The association between gut microbiota and serum hormones was analyzed with the R language.Results: Our findings reveal that the hyperandrogenic PCOS group exhibits lower richness and diversity of gut microbiota compared to the control group. Characteristic genera in PCOS patients with hyperandrogenism include Bifidobacterium, Enterobacteriaceae_unclassified, Streptococcus, Saccharimonadaceae, Enterococcus, and Eubacterium_nodatum_group. Five hormones, including 5β-androsterone, deoxycorticosterone, corticosterone, 11-dehydrocorticosterone, and cortexolone, emerge as potential serum biomarkers for identifying patients with hyperandrogenic-PCOS (HA-PCOS). Furthermore, a lower vitamin D3 level may act as a susceptibility factor, suggesting that vitamin D3 supplementation could serve as a potential intervention for PCOS with hyperandrogenism.Conclusion: Specific fecal microbiota and serum steroids may be used as characteristic markers for clinical diagnosis of hyperandrogenic-PCOS. This research enhances our understanding of the intricate interplay among hormones, gut microbiota, and hyperandrogenemia in patients with PCOS.

PURPOSE:The aim of this study was to investigate the efficacy and safety of early cumulus cell removal (ECCR) during human in vitro fertilization (IVF).METHODS:A retrospective analysis was performed between January 2011 and December 2019. The study enrolled 1131 couples who underwent IVF treatment with ECCR. After propensity score matching at a 1:1 ratio, 1131 couples who underwent overnight coincubation of gametes were selected. The main outcome measure was the cumulative live birth rate. Secondary outcome measures included the cumulative pregnancy rate, polyspermy rate, available embryo rate, miscarriage rate, malformation rate, time to live birth, and oocyte-to-baby rate.RESULTS:There were no significant differences found between the two groups in the polyspermy rate, available embryo rate, miscarriage rate, time to live birth, oocyte-to-baby rate, and neonatal congenital anomalies rate. The results of the study showed that ECCR was associated with a significantly higher cumulative live birth rate and cumulative pregnancy rate, along with a significantly lower fertilization rate.CONCLUSIONS:ECCR tended to confer increased cumulative live birth rate and had no negative effect on the neonatal malformation rate.

Research question:Does artificial oocyte activation (AOA) by a calcium ionophore (ionomycin) improve the previous fertilization failure or poor embryo development of intracytoplasmic sperm injection (ICSI) account for male factor infertility or other infertility causes?Design:This retrospective study involved 114 patients receiving ICSI-AOA in Shanghai First Maternity and Infant Hospital with previous ICSI fertilization failure or poor embryo development. The previous ICSI cycles of the same patients without AOA served as the control group. The fertilization rates, cleavage rates, transferable embryo rates and blastocyst formation rates of the two groups were compared. Additionally, the clinical pregnancy, implantation rate and live birth rates were also compared to assess the efficiency and safety of AOA. Furthermore, two subgroup analyses were performed in this study based on the cause of infertility and the reason for AOA. The fertilization rate, embryonic development potential and clinical outcome were compared among groups.Results:Among 114 ICSI-AOA cycles, the fertilization rate, top-quality embryo rate, implantation rate, clinical pregnancy per patient and live birth rate per patient were improved significantly compared with previous ICSI cycles (p<0.05 to P< 0.001), and the miscarriage rate in the AOA group was significantly lower than that of the control group (p<0.001). In the AOA subgroups based on the cause of infertility, the fertilization rates of each subgroup were significantly improved compared with previous control cycles except for the mixed factor infertility subgroup (p<0.05 to p<0.001). In the AOA subgroups based on the reason for AOA, the fertilization rates of each subgroup were significantly increased compared with those in their previous ICSI cycle without AOA (p<0.001); however, there was no significant difference in the top-quality embryo rate. No significant improvement was found in the implantation rates and the clinical pregnancy rate in each subgroup except for the poor embryo development subgroup. In the 114 AOA cycles, 35 healthy infants (21 singletons and 7 twins) were delivered without major congenital birth defects or malformations.Conclusion:This study showed that AOA with the calcium ionophore ionomycin can improve the reproductive outcomes of patients with previous fertilization failure and poor embryo development after ICSI.

Background Sperm cryopreservation is commonly used to preserve fertility but often decreases sperm quality by inducing oxidative stress. Pinus massoniana bark extract (Pinus massoniana, PMBE) exhibits strong antioxidant activity and has been used in traditional Chinese medicine. Objectives The objective was to determine whether adding PMBE to cryoprotectant could improve human sperm quality after cryo-resuscitation and to investigate the potential regulatory mechanisms. Methods PMBE was used as a cryoprotectant supplement. Hydrogen peroxide was applied to generate oxidative stress. The changes in sperm quality upon cryo-resuscitation and hydrogen peroxide treatment were measured by computer-aided sperm analysis and fluorescein isothiocyanate-labeled peanut (Pisum sativum) agglutinin staining. Oxidative stress was evaluated by dichlorofluorescein diacetate staining and the malondialdehyde content. The antioxidant capacity was determined by the glutathione peroxidase activity and the reduced glutathione content. Mass spectrometry was utilized to screen factors potentially regulated by PMBE, and the screened factors were evaluated by Western blot. Mouse in vitro fertilization was used to assess the effectiveness and safety of PMBE on the fertilization outcome of damaged spermatozoa. Results PMBE addition to cryoprotectant could effectively promote post-thaw vitality and acrosome reaction rates, reduce intracellular reactive oxygen species, and preserve antioxidant capacity and mitochondrial function in the presence of cryo-resuscitation and hydrogen peroxide treatment. Decreased in the expression of critical proteins such as inositol-3-phosphate synthase 1 (ISYNA1) and AKT1 upon resuscitation was rescued by PMBE supplementation, further reducing the cytoplasmic Nrf2 protein level and markedly increasing the nuclear level. PMBE dramatically enhanced the mRNA and protein expression of superoxide dismutase (SOD2). PMBE effectively alleviated the hydrogen peroxide-induced decline in the two-cell embryo rates. Conclusion Supplementing cryoprotectant with PMBE improves human sperm quality after cryo-resuscitation by reducing oxidative stress and stabilizing the functional proteins ISYNA1 and AKT1. PMBE addition improves the fertilization outcome of oxidative stress-damaged spermatozoa.

Background There is scarcity of information about the cumulative live birth rates(CLBRs) and time to live birth(TTLB) between progestin primed ovarian stimulation protocol(PPOS) and long GnRH agonist protocol. Objective To compare CLBRs and TTLB in women with normal ovarian reserve following PPOS with long GnRH agonist protocol. Methods A total of 995 women who underwent IVF using either PPOS (n=509) or GnRH antagonist (n=486) ovarian stimulation at the discretion of the attending physicians. The primary outcome measure was the CLBRs within 18 months from the day of ovarian stimulation. Results Both groups had almost comparable demographic and cycle stimulation characteristics except for duration of infertility which was shorter in the PPOS group. CLBRs after one complete IVF cycle including fresh and subsequent FET cycles within 18 months follow up were significantly lower in the PPOS group compared that in the long agonist group 206/509 (40.5%) and 307/486 (63.2%), respectively (odds ratio (OR): 0.641; 95% CI: 0.565-0.726). The average TTLB was significantly shorter in the long agonist group compared to the PPOS group (P < 0.01). In Kaplan-Meier analysis, the cumulative incidence of ongoing pregnancy leading to LB was significantly higher in the long agonist compared in the PPOS group (P < 0.001). Cox regression analysis revealed stimulation protocol adopted was strongly associated with the CLBRs after adjusting other confounding factors (OR =1.917 (1.152-3.190), P=0.012). Conclusion PPOS offers no advantage over conventional protocol in women with a normal ovarian reserve undergoing IVF. Keywords: PPOS, long GnRH agonist protocol, IVF, CLBRs, TTLB

To ascertain if assisted hatching (AH) increases the risk of placenta-associated diseases and perinatal outcomes after frozen–thawed cleavage-stage embryo transfer. We retrospectively evaluated 924 women who conceived with frozen–thawed cleavage-stage embryos transfer with (n = 390) or without (n = 534) laser-AH between 2013 and 2015. Data were obtained from the database on in vitro fertilization (IVF) patients in Shanghai First Maternity and Infant Hospital. We assessed neonatal (preterm birth, low birthweight, fetal macrosomia, stillbirth) and obstetric (miscarriage, ectopic pregnancy, post-term pregnancy, gestational diabetes (GDM), preeclampsia, intrahepatic cholestasis (ICP), placenta previa, placental abruption, premature rupture of membranes) outcomes. In twins, the median birthweight was lower in the AH group than that in the control group, and the prevalence of low birthweight (< 2500 g) was significantly higher in the AH group; after adjusting for maternal age, body mass index, mode of fertilization, and parity, no significant difference was found. In twins, no significant difference was detected in the prevalence of stillbirth or preterm pregnancy. In singleton births, there was no significant difference in the prevalence of low birthweight, macrosomia, preterm pregnancy or post-term pregnancy between the two groups. In singletons and twins, there were no significant differences in the prevalence of miscarriage, ectopic pregnancy, preeclampsia, GDM, ICP, or placenta abruption between the two groups. AH is a relatively safe method and our study provides important information for using this method in carefully selected patients.

目的 探讨不同受精方式治疗非男性因素不孕高龄女性患者的效果.方法 选取2017年1月至2019年12月同济大学附属第一妇婴保健院生殖医学中心606例非男性因素不孕高龄女性患者,根据受精方式分为常规体外受精(IVF)组(421例)和卵胞浆内单精子注射(ICSI)组(185例).比较两组的临床结局及子代出生情况.结果 两组受精率、囊胚形成率比较,差异有统计学意义(P<0.05);两组卵裂率、D3可用胚胎率、D3优质胚胎率、临床妊娠率、种植率比较,差异无统计学意义(P>0.05).两组孕龄、出生体重、出生率、低体重儿出生率、早产率比较,差异无统计学意义(P>0.05).结论 在非男性因素不孕高龄女性患者中,常规IVF治疗比ICSI可以获得更高的受精率和囊胚形成率,且ICSI治疗非男性因素不孕症并不能提高其种植率和出生率.

Background Previous studies of the effect of early cumulus cell removal (ECCR) on clinical outcomes remain controversial. Some studies indicated that ECCR combined early rescue ICSI contributed to avoid total fertilization failure, while the other studies demonstrated that ECCR may be detrimental to early embryo development. The aim of this study is to investigate the efficacy and safety of early cumulus cell removal (ECCR) during human IVF. Methods A retrospective analysis was performed between January 2011 and December 2016. The study enrolled 655 couples who underwent IVF treatments with ECCR. After propensity score matching at a 1:2 ratio, 1310 couples who underwent overnight coincubation of gametes were selected. All data were obtained from the Shanghai First Maternity and Infant Hospital IVF patient database. The main outcome measure was the live birth rate and the secondary outcome measures were the normal fertilization rate, polyspermy rate, available embryo rate, clinical pregnancy rate, miscarriage rate and malformation rate. Results No significant differences were found in the live birth rate (28.55% vs 28.4%; RR of 1.008; 95% CI: 0.869-1.170; p=0.916), clinical pregnancy rate (48.28% vs 45.16%; RR of 1.069; 95% CI: 0.951-1.202; p=0.268), implantation rate (32.67% vs 33%; p=0.896), miscarriage rate (13.33% vs 9.32%; RR of 1.43; 95% CI: 0.916-2.232; p=0.115), neonatal congenital anomalies rate (1.32% vs 1.01%; RR of 1.306; 95% CI: 0.315-5.417; p=0.713) or birthweight between the two groups. The study showed that ECCR was associated with a significantly lower fertilization rate (73.86% vs 80.12%; p=0.000), normal fertilization rate (2PN)(62.76% vs 69%, p=0.000) and available embryo rate (59.62% vs 62.29%, p=0.001). There were no significant differences in the polyspermy rate (11.10% vs 11.11%, p=0.982) and cleavage rate (93.93% vs 93.50%, p=0.279) between the ECCR group and traditional insemination group. Conclusions ECCR tended to confer increased risk of a lower available embryo rate but had no negative effect on the live birth rate or the neonatal malformation rate.

PURPOSE:Tubulin beta eight class VIII (TUBB8) is essential for oogenesis, fertilization, and pre-implantation embryo development in human. Although TUBB8 mutations were recently discovered in meiosis-arrested oocytes of infertile females, there is no effective therapy for this gene mutation caused infertility. Our study aims to further reveal the infertility-causing gene mutations in the patient's family and to explore whether the infertility could be rescued by optimizing the conditions of embryo culture and finally achieve the purpose of making the patient pregnant. METHODS:Whole-exome sequence analysis and Sanger sequencing were performed on patients' family members to screen and identify candidate mutant genes. Construction of plasmids, in vitro transcription, microinjection of disease-causing gene cRNA, and immunofluorescence staining were used to recapitulate the infertility phenotype observed in patients and to understand the pathogenic principles. Simultaneously, overexpression of mutant and wild-type cRNA of the candidate gene in mouse oocytes at either germinal vesicle (GV) or metaphase II (MII) stage was performed in the rescue experiment. RESULTS:We first identified a novel heritable TUBB8 mutation (c.1041C>A: p.N347K) in the coding region which specifically affects the first mitosis and causes the developmental arrest of early embryos in a three-generation family. We further demonstrated that TUBB8 mutation could lead to abnormal spindle assemble. And moreover, additional expression of wild-type TUBB8 cRNA in the mouse oocytes in which the mutant TUBB8 were expressed can successfully rescue the developmental defects of resulting embryo and produce full-term offspring. CONCLUSIONS:Our study not only defines a novel mutation of TUBB8 causing the early cleavage arrest of embryos, but also provides an important basis for treating such female infertility in the future.

Poor oocyte quality is associated with early embryo developmental arrest and infertility. Maternal gene plays crucial roles in the regulation of oocyte maturation, and its mutation is a common cause of female infertility. However, how to improve oocyte quality and develop effective therapy for maternal gene mutation remains elusive. Here, we use Zar1 as an example to assess the feasibility of genome transfer to cure maternal gene mutation–caused female infertility. We first discover that cytoplasmic deficiency primarily leads to Zar1-null embryo developmental arrest by disturbing maternal transcript degradation and minor zygotic genome activation (ZGA) during the maternal-zygotic transition. We next perform genome transfer at the oocyte (spindle transfer or polar body transfer) and zygote (early pronuclear transfer or late pronuclear transfer) stages to validate the feasibility of preventing Zar1 mutation–caused infertility. We finally demonstrate that genome transfer either at the oocyte or at the early pronuclear stage can support normal preimplantation embryo development and produce live offspring. Moreover, those pups grow to adulthood and show normal fertility. Therefore, our findings provide an effective basis of therapies for the treatment of female infertility caused by maternal gene mutation.

目的 评估枸橼酸托瑞米芬联合维生素治疗特发性男性少弱精子症的疗效.方法 门诊病人60例随机分成观察组和对照组.观察组:口服托瑞米芬、维生素C和天然维生素E;观察组:口服枸橼酸他莫昔芬、维生素C和天然维生素E.观察指标包括:精液常规分析和内分泌指标(FSH,LH,T).结果 治疗3个月后观察组和对照组的精子浓度与治疗前比较均有提高,差异均有统计学意义(P＜0.05);治疗后两组间比较,差异无统计学意义(P＞0.05).前向运动精子百分率观察组较治疗前有提高,差异有统计学意义(P＜ 0.05);对照组比较治疗前有提高,但差异无统计学意义(P＞0.05);治疗后两组间比较,差异有统计学意义(P＜0.05).观察组与对照组的3项内分泌指标治疗后与治疗前比较均有上升,差异均有统计学意义(P＜0.05).结论 托瑞米芬联合维生素治疗特发性少弱精子症有一定效果.

目的 比较不同精子DNA碎片指数(DNA fragmentation index,DFI)的精子冷冻前后的浓度、活力变化及复苏率,明确DFI对精子冷冻的影响.方法 2017年在上海市第一妇婴保健院生殖中心行体外受精-胚胎移植(in vitrofertilization and embryo transfer,IVF-ET)的患者中,自精冻精患者共55例,男方通过病史询问、生殖系统超声检查、染色体检查排除遗传及其他因素,入组排除因严重少弱精子症和隐匿精子症冻精的患者,入组患者36例,按DFI≥27％和DFI＜27％分为两组.比较两组患者精子冷冻前后的精子浓度、活力差以及复苏率.结果 36例患者分为DFI≥ 27％组1 1例,DFI＜27％组25例.两组间年龄、不育年限和精子形态率等一般情况比较差异无统计学意义(P=0.29;P=0.77;P=0.28).两组间精子冷冻前后的浓度变化、活力变化及前向运动精子(PR)总数变化的差异均无统计学意义(P=0.07;P=0.76;P=0.58),比较两组冷冻复苏率差异无统计学意义(P=0.52).结论 不同DNA碎片率的精子冷冻解冻后的浓度、活力变化以及冷冻复苏率并没有显著差异,DFI异常不是精子冷冻的禁忌证.

STUDY QUESTION:Does brief incubation of oocytes and spermatozoa improve the live birth rate (LBR) of IVF when compared with that of standard incubation? SUMMARY ANSWER:Brief incubation of gametes does not improve the LBR of IVF when compared with standard incubation. WHAT IS KNOWN ALREADY:Some small randomized studies showed that brief incubation was associated with a significantly higher ongoing pregnancy rate than standard incubation. STUDY DESIGN, SIZE, DURATION:This is a randomized triple blind study of 320 infertile women for their first or repeated cycles undergoing IVF between September 2015 and October 2016. PARTICIPANTS/MATERIALS, SETTING, METHODS:Women were randomized into the brief incubation group (n = 160) or the standard incubation group (n = 160) according to a computer-generated randomization list. Oocytes were incubated with spermatozoa (0.3-1.2 million motile sperm/ml) for 3-4 h in the brief incubation group while oocytes were incubated with spermatozoa at similar concentration for 20 h in the standard incubation group. The primary outcome was the LBR (a baby born alive after 22 weeks gestation) in the fresh cycle. MAIN RESULTS AND THE ROLE OF CHANCE:There was no significant difference in the LBR between the brief and standard incubation groups based on both intention-to-treat [33.0% (53/160) versus 36.8% (59/160), relative risk (RR) 0.898 (95% CI = 0.666-1.212), P = 0.482] and per protocol [41.4%(53/128) versus 41.0% (59/144), RR1.011 (95% CI = 0.760-1.343), P = 0.942] analyses. Clinical pregnancy, ongoing pregnancy, miscarriage, multiple pregnancy and implantation rates were comparable for the two groups. Similar results were found with subgroup analysis of advanced maternal age, abnormal semen analysis and repeated IVF cycles. No differences were observed in cumulative LBR between two groups. LIMITATIONS, REASONS FOR CAUTION:Various motile sperm concentrations of 0.3-1.2 million per ml were used for insemination and the reactive oxygen species level in the insemination medium was not measured. The highest level at 1.2 million per ml is still relatively low compared to prior studies, therefore we do not know whether brief incubation can improve the LBR using higher concentrations of spermatozoa. The present sample size may not be adequate to detect a smaller difference in the LBR. WIDER IMPLICATIONS OF THE FINDINGS:The present study demonstrated that a brief incubation of gametes had no significant beneficial effect on the LBR when compared with the standard incubation. The practice of brief incubation of gametes is not necessary and this can save the already tight manpower in many laboratories. STUDY FUNDING/COMPETING INTERESTS:The study was supported by the Merck-Serono China Research Fund for Fertility Experts (2015), which was not involved in study design, execution, data analysis and manuscript preparation. There are no conflicts of interest for all authors. TRIAL REGISTRATION NUMBER:ClinicalTrials.gov Identifier NCT02534857. TRIAL REGISTRATION DATE:28 August 2015. DATE OF FIRST PATIENT’S ENROLMENT:8 September 2015.

Oxidative stress is associated with functional disorder of trophoblast cells. Our previous studies have demonstrated that cyclosporin A (CsA) promotes the activity of normal human trophoblast cells. We further investigated the role and mechanism of CsA on oxidative stress in trophoblast cells. JEG-3 cells were co-cultured with H2O2 and CsA. Cell viability and morphology were measured by MTT assay and inverted microscope. Reactive oxygen species (ROS) was analyzed by fluorescence microscopy. Cell mitochondrial membrane potential (MMP) was determined by flow cytometric analysis. Malondialdehyde (MDA) production, superoxide dismutase (SOD) and catalase (CAT) activities were examined using colorimetric assays. The expression and phosphorylation of FAK and Src kinase proteins were examined by western blotting. CsA increased JEG-3 cell viability and reduced the morphologic injury induced by H2O2 treatment. CsA decreased ROS and MDA production, increased SOD and CAT activities, and restored the MMP of H2O2 treated JEG-3 cells. CsA administration suppressed H2O2-induced reduction of FAK and Src phosphorylation. Blocking the activation of FAK or Src attenuated the protective effect of CsA on JEG-3 cells in H2O2-induced oxidative injury. CsA protects JEG-3 cells from H2O2-induced oxidative injury, and the FAK/Src signaling pathway plays an important role in this process.

To compare the pregnancy outcomes of in vitro fertilisation (IVF) following tubal occlusion by microcoil placement versus laparoscopic tubal ligation for hydrosalpinges. This was a single centre retrospective study of 127 infertile women aged <43 years with unilateral or bilateral hydrosalpinges on transvaginal ultrasound who underwent either tubal occlusion by the microcoil placement under X-ray control (the microcoil group (n = 60)) or laparoscopic tubal ligation (the ligation group (n = 67)) prior to IVF at the discretion of the attending physicians. In all women, laparoscopy was not considered to be contraindicated due to extensive pelvic adhesions. The pregnancy outcomes of the IVF cycle following the treatment for hydrosalpinges were compared. Both groups had comparable demographic and cycle stimulation characteristics. The positive pregnancy test (43.3% (26/60) vs 64.2% (43/67), p = 0.02, RR = 0.68 (0.48-0.95)), ongoing pregnancy (35.0% (21/60) vs 58.2% (39/67), p = 0.009, RR = 0.60 (0.40-0.89)) and implantation rates (33.3% (34/102) vs 49.5% (56/113), p = 0.016, RR 0.67 (0.48-0.94)) were significantly lower in the microcoil group than those in the ligation group. Both groups had similar miscarriage, multiple pregnancy and ectopic pregnancy rates. A multivariate logistic regression revealed that only the women's age and the treatment method of hydrosalpinx were significant factors in predicting the ongoing pregnancy. A lower ongoing pregnancy rate of IVF was found in women with hydrosalpinges following tubal occlusion by microcoil placement under X-ray when compared with laparoscopic tubal ligation.

Objective:To study the effect of smooth endoplasmic reticulum aggregation (SERa)in oocytes on oocyte-to-embryo rate.Methods:The data of 250 cycles of ICSI-treated patients from January 2013 to December 2014 were retrospectively analyzed.According to whether oocytes had SERa,the cycles were divided into two groups:at least one oocyte with SERa positive in SERa+ group(group A,50 cycle)and all oocytes without SERa in SERa-group(group B,200 cycle).The fertilization rate,abnormal fertilization rate,cleavage rate,available embryos rate and oocyte-to-embryo rate were compared between the two groups.Results:The significantly lower cleavage rate,available embryos rate,oocyte-to-embryo rate and significantly higher multiple pronucleus fertilization rate were observed in the SERa+ cycles as compared to SERa-cycles(P＜0.05).SERa+ cycles had significantly lower cleavage rate,available embryos rate & oocyte-to-embryo rate,and significantly higher multiple pronucleus fertilization rate (P ＜ 0.05) compared with SERa oocyes.Among SERa+ cycles,SERa+ oocytes had higher multiple pronucleus fertilization rate compared to SERa oocytes(P＜0.05),while there was no significant difference in cleavage rate,available embryos rate and oocyte-to-embryo rate between SERa+ oocytes and SERa-oocytes(P＞0.05).Conclusions:Oocyte cytoplasmic smooth endoplasmic reticulum aggregation increases abnormal fertilization rate and reduces oocyt-to-embryo rate.

Background: Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. An lncRNA, namely, Prader-Willi region nonprotein coding RNA 2 (PWRN2), was up-regulated in the cumulus cells of patients with PCOS. However, the molecular mechanism of PWRN2 in PCOS remains largely unknown. Methods: In this study, the expression levels of PWRN2 were tested in cumulus cells through qRT-PCR analysis to confirm its potential roles in oocyte nuclear maturation of PCOS. A PWRN2-mediated ceRNA network was constructed based on three microarray datasets to investigate the molecular mechanism of PWRN2 in oocyte development of patients with PCOS. The direct interactions of the candidate genes of the ceRNA network were also demonstrated by dual-luciferase reporter assay. Results: PWRN2 was found to be associated with oocyte nuclear maturation in patients with PCOS in contrast to that in normal patients. Based on the microarray data, 176 lncRNAs (118 up-regulated and 58 down-regulated) and 131 mRNAs (84 up-regulated and 47 down-regulated) were identified to be regulated by PWRN2. A PWRN2-miR-92b-3p-TMEM120B ceRNA network was constructed based on results of analysis of the combined three microarray datasets (lncRNA+mRNA microarray in KGN/shPWRN2 in this study, miRNAs microarray and lncRNA+mRNA microarray in PCOS cumulus cells reported in previous studies). The coexpression characteristics of the genes (PWRN2, miR-92b-3p and TMEM120B) were detected in the cumulus cells of cumulus-oocyte complexes at different nuclear maturity stages in PCOS. These results are in accordance with the ceRNA hypothesis. Moreover, luciferase activity assay revealed that miR-92b-3p directly binds to PWRN2 and targets TMEM120B. Conclusions: PWNR2 plays important roles in oocyte nuclear maturation in PCOS by functioning as a ceRNA to reduce the availability of miR-92b-3p for TMEM120B target binding during oocyte maturation in PCOS. Our findings would provide new information and clarify abnormal oocyte development in PCOS.