目的 研究授精前剥除部分卵丘细胞的卵母细胞的受精、胚胎发育和临床结局. 方法 授精前用机械法剥除卵母细胞外包裹的部分卵丘细胞,每个卵母细胞采用微滴法单独受精(A组).对照组(B组)采用常规体外受精-胚胎移植(IVF-ET)受精方式进行.比较两组受精率(2PN、1PN和3PN)、卵裂率(2PN、1PN和3PN)、优胚率、临床妊娠率、种植率和流产率. 结果 A组和B组的受精率分别是76.51％和73.74％(2PN受精率分别是60.64％,61.62％;1PN受精率分别是1.61％,0.86％；3PN受精率分别是10.44％,9.69％)、卵裂率分别是98.95％和99.17％(2PN卵裂率分别是97.02,98.01％;1PN卵裂率分别是100％,85.71％；3PN卵裂率分别是100％,100％)、优质胚胎率分别是60.75％,62.68％、临床妊娠率分别是63.89％,61.76％、种植率分别是48.00％,38.78％、流产率分别是9.09％,11.90％.所有指标均无统计学意义. 结论 卵母细胞授精前剥除部分卵丘细胞不会影响IVF-ET的临床结局,为利用卵丘细胞进行非侵入性种植前遗传学诊断提供临床依据.

目的 探讨废弃胚胎作为体外受精(IVF)实验室质控的可行性. 方法 通过囊胚序贯培养将无原核(0PN)、单原核(1PN)、3原核(3PN)和发育迟缓或停滞的2原核(2PN)的废弃胚胎进行囊胚培养,比较不同来源胚胎的囊胚形成情况. 结果 共收集396个废弃胚胎,经序贯培养形成98个囊胚(24.75％),0PN、2PN、1PN、3PN的囊胚形成率分别为50.50％(51/101)、24.59％(30/122)、17.07％(7/41)、7.58％(10/132),其中0PN、停滞的2PN的胚胎有孵出,孵出率分别为47.06 ％(24/51)和33.33％(10/30)；有囊胚形成与无囊胚形成患者的临床妊娠率分别为30.28％(43/142)、19.72％％(28/142),两者相比有统计学差异(P＜0.05)；特别是0PN+2PN患者的临床妊娠率为23.24％(33/142),显著高于无囊胚形成者的9.15％(13/142)(P＜0.01). 结论 废弃胚胎有不同程度的发育潜能,可部分发育成囊胚,特别是0PN+2PN囊胚形成率超过35％,可以作为IVF实验室质控的一种辅助方法,并可作为临床妊娠结局预测的参考指标.

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. The abnormalities of endocrine and intra-ovarian paracrine interactions may change the microenvironment for oocyte development during the folliculogenesis process and reduce the developmental competence of oocytes in PCOS patients who are suffering from anovulatory infertility and pregnancy loss. In this microenvironment, the cross talk between an oocyte and the surrounding cumulus cells (CCs) is critical for achieving oocyte competence. The aim of our study was to investigate the gene expression profiles of CCs obtained from PCOS patients undergoing IVF cycles in terms of oocyte maturation by using human Genome U133 Plus 2.0 microarrays. A total of 59 genes were differentially expressed in two CC groups. Most of these genes were identified to be involved in one or more of the following pathways: receptor interactions, calcium signaling, metabolism and biosynthesis, focal adhesion, melanogenesis, leukocyte transendothelial migration, Wnt signaling, and type 2 diabetes mellitus. According to the different expression levels in the microarrays and their putative functions, six differentially expressed genes (LHCGR, ANGPTL1, TNIK, GRIN2A, SFRP4, and SOCS3) were selected and analyzed by quantitative RT-PCR (qRT-PCR). The qRT-PCR results were consistent with the microarray data. Moreover, the molecular signatures (LHCGR, TNIK, and SOCS3) were associated with developmental potential from embryo to blastocyst stage and were proposed as biomarkers of embryo viability in PCOS patients. Our results may be clinically important as they offer a new potential strategy for competent oocyte/embryo selection in PCOS patients.

BACKGROUND:Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. The developmental competence of oocytes and embryos in PCOS patients is reduced to a certain extent (comparing to non-PCOS patients, the high quality embryo rate was decreased by 16% from the data of our centre) during the in vitro fertilization (IVF) process. Cross-talk between the oocyte and cumulus cells is critical for oocyte maturation and embryo competence. In this study, we have evaluated the transcription of specific genes in cumulus cells harvested from pre-ovulatory follicles of PCOS patients before IVF, according to individual oocyte nuclear maturity and developmental competence. Seven genes (RUNX2, PSAT1, ADAMTS9, CXCL1, CXCL2, CXCL3, and ITGB5) were targeted from our previous cDNA microarray data which isolated genes related to oocyte nuclear maturation in PCOS patients. Two additional genes which had been found to be associated with oocyte maturation or embryo quality in non-PCOS patients (GPX3 and PTX3) were also studied.METHODS:The mRNA expression levels of cumulus cells were detected by qRT- PCR.RESULTS:Consistent with our previous cDNA microarray data, with the exception of GPX3 and PTX3, the selected 7 genes were related to oocyte nuclear maturation in PCOS patients. Noticeably, the expression level of RUNX2 was lower in cumulus cells derived from oocytes that could develop into blastocysts than the level of expression from oocytes that could not. The PTX3 expression level was significantly lower in cumulus cells from oocytes with two normal pronuclei than that from oocytes that formed >2 pronuclei (MPN) after fertilization. GPX3 mRNA levels were decreased in cumulus cells isolated from oocytes that developed into blastocysts with high potential development competence.CONCLUSIONS:Several cumulus cell genes were associated with oocyte maturation, fertilization and embryo quality in PCOS patients. RUNX2 and GPX3 are candidate genetic markers in the monitoring of embryo quality for PCOS patients, whereas PTX3 mainly played a role in fertilization process. Together with morphological evaluation, cumulus cells genes may serve as biomarkers of oocyte and embryo selection during the IVF process for PCOS patients and may advance our understanding of PCOS.

目的 通过对常规体外授精(IVF)失败(未观察到第二极体)后不同时间行补救性卵细胞浆单精子显微注射(ICSI)的临床资料比较,探讨补救性ICSI的最佳时间. 方法 回顾性分析常规体外授精(IVF)失败后6 h(早期组)与22 h(传统组)行补救性ICSI的临床资料,比较两组的受精率、卵裂率、优胚率、种植率和妊娠率. 结果 早期组受精率、种植率、临床妊娠率分别为74.6%(56/75)、37.5%(9/24)、53.8%(7/13);传统组受精率为63.0%(29/46)、种植率和临床妊娠率均为0.早期组7例妊娠中4例单胎,3例流产;早期组和传统组受精率比较,差异无统计学意义,妊娠率和种植率比较,差异有统计学意义(P<0.05); 结论 早期补救性ICSI在受精率、种植率和临床妊娠率上可能优于传统补救性ICSI.