Background Phthalate exposures have been shown to be inversely associated with reproductive success among women undergoing assisted reproductive technology (ART). However, the underlying mechanisms are unknown. Objectives To explore blood coagulation function as the mediating role of associations between exposure to phthalates and ART outcomes. Methods A total of 735 women from the Tongji Reproductive and Environmental (TREE) study were included. Urine samples collected at recruitment were quantified for 8 phthalate metabolites, and blood clotting time and platelet indices were also determined. Generalized linear regression, logistic regression, weighted quantile sum (WQS) regression, or Bayesian kernel machine regression (BKMR) models were applied to investigate the associations among individual and mixture of phthalate metabolites, blood coagulation parameters, and ART outcomes. The mediation role of blood coagulation parameters was estimated by mediation analysis. Results Mono-n-butyl phthalate (MBP), mono-isobutyl phthalate (MiBP), monobenzyl phthalate (MBzP), mono(2-ethyl-5-carboxypentyl) phthalate (MECPP), and molar sum of di(2-ethylhexyl) phthalate metabolites (∑DEHP) were positively associated with platelet indices. Phthalate metabolite mixture was also positively associated with platelet count (PLT), mean platelet volume (MPV), and plateletcrit (PCT), whereas inversely associated with international normalized ratio (INR). Meanwhile, PLT and PCT were inversely associated with the odds of implantation success and live birth, while prothrombin time and INR were positively associated with the odds of implantation success. Mediation analyses showed indirect effects of above-mentioned phthalate metabolites and phthalate mixture on the odds of implantation success and live birth through PLT or PCT, with the proportion mediated ranging from 3.44% to 8.96%. Conclusions Phthalates may increase the risks of ART failure through enhancing blood coagulation function. More studies are warranted to verify the findings.

Mono-butyl phthalate (MBP), the metabolite of dibutyl phthalate (DBP), is the most abundant phthalate metabolite found in Chinese women. Extracellular vesicles (EVs) are nanoscale lipid bilayer particles produced by extensive kinds of cells, serving a key role in intercellular communication. Extracellular vesicle miRNAs (EV-miRNAs) in follicular fluid (FF) have been evidenced to be associated with female reproductive health. The objective of this study was to investigate the associations of EV-miRNAs expressed profile with DBP exposure in FF of female participants and expose its potential mechanism in impaired oocyte development. Based on participants’ FF MBP concentrations and fertilization status, we compared the miRNA expression between the FF-EVs of group A (high DBP exposure and impaired fertilization) and group B (low DBP exposure and normal fertilization). Compared with group B, miR-1246, miR-3679-5p, miR-423-5p, miR-5585-3p, miR-116-5p, miR-172-5p were upregulated, while miR-34b-3p was downregulated in group A. Target genes of the differently expressed miRNAs were predicted, and the functional analysis was performed. Furthermore, we exposed human ovarian granulosa tumor cell line (KGN) to MBP (4ug/L) to isolate the EVs from the culture medium and validated the expression levels of different miRNAs. We found that MBP exposure was significantly associated with increased levels of miR-116-5p (P = 0.01). In addition, we demonstrated that the most different miRNA, miR-116-5p regulated oocyte fertilization by inhibiting FOXO3a. Our findings suggested that EV-miRNAs in the FF might mediate MBP toxicity in oocytes.

IntroductionPersonal care products (PCPs) contain a number of endocrine-disrupting chemicals (EDCs) that could potentially affect the reproductive function in women of childbearing age. However, studies focused on the effects of PCPs use on reproductive outcomes are very limited. The current study aimed to explore the relationships between PCPs use patterns and reproductive outcomes in women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment.MethodsA total of 1500 women from the Tongji Reproductive and Environmental (TREE) study between December 2018 and January 2020 were included in this study. Participants provided characteristics of PCPs use within the previous three months. Retrieved oocyte number, mature oocyte number, two distinct pronuclei (2PN) zygote number, fertilization rate, cleavage rate, blastocyst formation rate, implantation, clinical pregnancy, miscarriage, and live birth were followed up as reproductive endpoints. Generalized linear regression model was utilized to assess the associations between various categories of PCPs use and reproductive endpoints of IVF/ICSI.ResultsAfter adjusting for relevant covariates, women who used skin care products ≥14 times per week had a reduction of 22.4% in the maturation rate (95% CI: -39.2%, -1.6%) compared to participants who did not use skin care products. After transferring fresh embryos, women who used cosmetics 1–2 times per week (adjusted OR = 2.2, 95% CI: 1.0, 4.8) or 3–7 times per week (adjusted OR = 2.5, 95% CI: 1.2, 5.2) had a higher possibility of miscarriage than those who did not use cosmetics. There was negative association between the use of gel or soap and the cleavage rate among women aged < 30 years old (P for interaction = 0.01). Among women with BMI ≥ 24 kg/m2, the use of gel or soap was negatively associated with the blastocyst formation rate (P for interaction = 0.04), while cosmetics use was negatively associated with the maturation rate (P for interaction = 0.001).ConclusionOur findings suggest that the use of PCPs in women of reproductive age have a potential adverse impact on IVF/ICSI outcomes, particularly skin care and cosmetic products.

Di(2-ethylhexyl) phthalate (DEHP) is a widely recognized environmental endocrine disruptor that potentially impacts female reproductive function, although the specific mechanisms leading to such impairment remain unclear. A growing body of research has revealed that the endoplasmic reticulum and mitochondrial function significantly influence oocyte quality. The structure of mitochondria-associated endoplasmic reticulum membranes (MAMs) is crucial for facilitating the exchange of Ca2+, lipids, and metabolites. This study aimed to investigate the alterations in the composition and function of MAMs after DEHP exposure and to elucidate the underlying mechanisms of ovarian toxicity. The female mice were exposed to DEHP at doses of 5 and 500 mg/kg/day for one month. The results revealed that DEHP exposure led to reduced serum anti-Müllerian hormone levels and increased atretic follicles in mice. DEHP induced endoplasmic reticulum stress and disrupted calcium homeostasis in oocytes. Furthermore, DEHP impaired the mitochondrial function of oocytes and reduced their membrane potential, and promoting apoptosis. Similar results were observed in human granulosa cells after exposure to mono-(2-ethylhexyl) phthalate (MEHP, metabolites of DEHP) in vitro. Proteomic analysis and transmission electron microscopy revealed modifications in the functional proteins and structure of the MAMs, and the suppression of oxidative phosphorylation pathways. The findings of this investigation provide a new perspective on the mechanism underlying the reproductive toxicity of DEHP in females.

Phthalates are a class of environmental endocrine disrupting chemicals which can cause reproductive system damages. However, data about reproductive toxicity spectrum of phthalate metabolites among Chinese women undergoing in vitro fertilization (IVF) treatments are scarce yet. Previous studies regarding underlying embryo toxicities focused on oxidative stress and apoptosis, while energy metabolism abnormality might be another key cause for embryo developmental disruptions. Here, we found that among the measured eight phthalate metabolites, monomethyl phthalate (MMP) had the second highest urinary concentration in women receiving IVF. Compare to the lowest exposure level group, MMP in tertile 3 was associated with fewer counts of oocyte retrieved and good-quality embryos, and MMP in tertile 2 was correlated with reduced good-quality embryo rate. The direct embryo toxicities of MMP were studied using mouse 2-cell embryos. Consistent to results found in human populations, exposure to MMP induced mouse early embryo developmental delay. Furthermore, MMP exposure led to excessive reactive oxygen species production in early embryos, and antioxidant can partially rescue the early embryo development slow down. Embryo apoptosis could also be caused by oxidative stress. To be noted, elevated apoptosis level was not found in live "slow" embryos but dead embryos, which suggested that apoptosis was not related to early embryo developmental delay. Additionally, MMP exposure depleted adenosine triphosphate (ATP) synthesis of early embryos, which could be reversed by antioxidant. In conclusion, MMP, as the newly found embryonic toxicant in Chinese women, resulted in early embryo development delay, apoptosis, and energy metabolism disruptions via inducing redox imbalance.

RESEARCH QUESTION:Are blastocysts derived from in-vitro-matured metaphase I (MI) oocytes less likely to produce usable embryos for transfer compared with those derived from in-vivo-matured oocytes in cycles undergoing preimplantation genetic testing (PGT)? DESIGN:The primary outcome was usable blastocyst rate, which was compared between blastocysts derived from in-vitro-matured MI oocytes after ovarian stimulation and from in-vivo-matured oocytes. Logistic regression analysis using generalized estimating equations was used to control for confounders in the analysis of factors that may influence the chance of a blastocyst being usable and in the comparison of embryological outcomes. Student's t-test, Mann-Whitney U test, chi-squared tests or Fisher's exact tests were used to compare clinical and pregnancy outcomes. RESULTS:A total of 1810 injected metaphase II (MII) oocytes from 154 PGT cycles involving 154 couples were included in this study. A total of 1577 MII oocytes were in-vivo-matured and 233 were in-vitro-matured MI oocytes. The usable blastocyst rate was similar between the in-vitro-matured MI oocyte group and the in-vivo-matured oocyte group (adjusted RR 0.97, 95% CI 0.40 to 2.34). Three live births were achieved using usable blastocysts derived from in-vitro-matured MI oocytes. CONCLUSIONS:If in-vitro-matured MI oocytes can be fertilized and develop into blastocysts, their ability to provide usable embryos for transfer is similar compared with those developed from in-vivo-matured oocytes. These blastocysts could be considered valuable for women with few viable embryos in assisted reproductive technology cycles.

目的 通过高通量测序技术(NGS)探讨D3胚胎卵裂球数目与胚胎发育潜能和整倍性之间的关系.方法 回顾性分析2018年1月至2021年12月于本院生殖中心行胚胎植入前非整倍体遗传学检测(PGT-A)助孕的261个治疗周期共920枚活检囊胚的数据资料,根据D3胚胎卵裂球数目不同进行分组(4细胞组、5细胞组、6细胞组、7细胞组、8细胞组、9细胞组、10细胞组及≥11细胞组),评估各组后续获得优质囊胚和整倍体囊胚的机会,并统计分析各组间临床结局的差异.结果 (1)与8细胞组相比,5细胞、6细胞、7细胞及10细胞组优质囊胚率显著降低(P=0.00),调整女方年龄因素以后,差异仍有显著性(P=0.00);卵裂球数目合并分组后,≤7细胞组、9～10细胞组的优质囊胚率显著低于8细胞组,调整年龄因素以后,差异仍有显著性(P＜0.05);≥11细胞组优质囊胚率与8细胞组间无显著性差异(P=0.51),调整女方年龄后,仍无显著性差异(P=0.45).(2)与8细胞组相比,其他各组囊胚的整倍体率均无显著性差异(P＞0.05);调整年龄后,各组间囊胚整倍体率仍无显著性差异(P＞0.05);卵裂球数目合并分组后,各组间随着卵裂球发育速度增快,囊胚整倍体率增加,但差异尚无统计学意义(P＞0.05);调整年龄混杂因素后,各组间的囊胚整倍体率仍无显著性差异(P＞0.05).(3)随着卵裂球数目增加,各组临床妊娠率、活产率呈上升趋势;卵裂球数目合并分组后,≥11细胞组的临床妊娠率和活产率最高,≤7细胞组的临床妊娠率和活产率最低,但差异均尚无统计学意义(P＞0.05).结论 D3卵裂期胚胎的发育潜能随着卵裂球数目的增加而增加,≥11细胞的D3快速分裂胚胎具有同8细胞胚胎相似的整倍体率和活产率,为临床单胚胎移植胚胎选择策略提供一定的参考.

辅助生殖技术(assisted reproductive technology, ART)的最终目标是获得健康单胎活产儿.然而,在临床确认的妊娠中依然有10％ ～15％的患者以流产告终,染色体异常约占早期妊娠丢失的 50％ [1-2].研究表明, ART中基于形态学评分的高质量胚胎很大一部分是非整倍体[3] .随着我国"三孩政策"全面实施,出生人口数、高龄、高危人群数随之增加,预防出生缺陷、提高出生缺陷防控水平迫在眉睫.近年来,胚胎植入前遗传学检测(preimplantation genetic testing,PGT)技术通过对植入前胚胎进行染色体及遗传致病基因的筛查,选择染色体无异常胚胎植入到宫腔内,从而在提高妊娠率、降低流产率和减少出生缺陷等方面有一定的临床价值.目前PGT 主要分为非整倍体 PGT( PGT for aneuploidy, PGT-A)、单基因病 PGT ( PGT for monogenic/single gene defects,PGT-M)和染色体结构重排 PGT(PGT for chromosomal structural rearrangements, PGT-SR).通过PGT后的胚胎通常比较珍贵,如何合理利用胚胎,改善PGT人群临床妊娠率、减少出生缺陷,一直是人们关注的焦点.本文重点从胚胎选择策略、胚胎移植方案、遗传咨询和产前诊断等方面进行综述.

目的 观察新鲜卵胞浆内单精子注射(ICSI)移植周期D3胚胎卵裂球数目不同对妊娠结局和新生儿结局的影响,以期优化单胚胎选择策略.方法 回顾性分析2018年1月至2020年12月在本中心行ICSI治疗的2 618个新鲜移植周期的临床资料,按照第三天(D3)胚胎卵裂球数目分为6组:≤6细胞组(78例)、7细胞组(199例)、8细胞组(1 936例)、9细胞组(178例)、10细胞组(163例)和≥11细胞组(64例),比较组间妊娠结局和新生儿结局.结果 妊娠结局的比较:8细胞组的临床妊娠率(52.8％)显著高于≤6细胞组(20.5％)、7细胞组(34.2％)和9细胞组(34.3％),活产率(44.5％)也显著高于≤6细胞组(11.5％)、7细胞组(27.6％)和9细胞组(28.7％),差异均有统计学意义(P＜0.001);10细胞组和≥11细胞组的临床妊娠率和活产率略低于8细胞组,差异尚无统计学意义(P＞0.05).新生儿结局的比较:7细胞组的低出生体重儿占比显著高于8细胞组(10.9％vs.2.7％,P＜0.001),组间其他新生儿结局指标差异均无统计学意义(P＞0.05);≤6细胞组获得9例健康活产儿.结论 新鲜ICSI移植周期中,优先考虑移植D3的8细胞胚胎,其次为10个及以上细胞的胚胎;9细胞胚胎和7细胞胚胎具有相似的临床结局,但7细胞胚胎低出生体重儿风险增加;当没有优质胚胎可供移植时,≤6细胞胚胎依然有健康活产的机会.