Objective To investigate the malformation of infants born in fresh cycle and melt cycle of assisted reproductive technology.Methods 35 098 infertile patients underwent test-tube cycle transplantation,15 049 cases of fresh embryo transplantation were selected as control group,and 20 049 cases of freeze-thaw transplantation were selected as observation group.The fetal malformations of two groups and different types of fetal malformations were compared.Results There were 88 cases of fetal malformation in the control group,the malformation rate was 0.58％.In the observation group,there were 113 cases of fetal malformation,the rate of fetal malformation was 0.56％.There was no significant difference in the rate of fetal malformation between the two groups(P＞0.05).There was no significant difference in the rates of craniocerebral nerve,eye,ear,facial and neck,cardiovascular circulation,musculoskeletal,genitourinary system,chromosome diseases,digestive system malformations and complex malformations between the two groups(P＞0.05).Conclusion During the implementation of assisted reproductive technology,embryo freezing and thawing transfer will not increase the rate of birth defects,and frozen and thawing embryo transfer technology is safe.

Objective:To explore the clinical application value of short-term insemination combined with early rescue intracytoplasmic sperm injection(R-ICSI). Methods:The clinical data of patients who received IVF/ICSI treatment in the Reproductive Hospital of Guangxi Zhuang Autonomous Region from January 2020 to December 2021 were retrospectively analyzed.According to the method of insemination,the patients were divided into R-ICSI group(105 cycles),conventional IVF group(2 058 cycles)and conventional ICSI group(486 cycles).There were 56 fresh transplant cycles in the R-ICSI group,1 059 fresh transplant cycles in the IVF group and 259 fresh transplant cycles in ICSI transplant cycles.The basic information,laboratory outcome,and pregnancy outcome were analyzed among the three groups. Results:In the comparison of basic information,there were no significant differences in female age,BMI,years of infertility,Gn dosage and days,basal sex hormone levels,average number of oocytes retrieved,and transplantable cycle rate among the three groups(P＞0.05).The primary infertility rate in the R-ICSI group and ICSI group was significantly higher than that in the IVF group(P＜0.05),but there was no significant difference between the R-ICSI group and ICSI group(P＞0.05).No patients in the R-ICSI group used follicular phase long protocol,while there was no significant difference in the proportion of cycles used follicular phase long protocol for ovulation induction between the IVF group and the ICSI group(P＞0.05).The proportion of cycles used luteal phase long protocol for ovulation induction was significantly higher than that of the IVF group(P＜0.05),but there was no significant difference compared to the ICSI group(P＞0.05).There was no significant difference in the proportion of cycles used super-long protocol and antagonist protocol for ovulation induction among the three groups(P＞0.05).In the comparison of laboratory outcomes,the total fertilization rate and normal fertilization rate of patients in the R-ICSI group and ICSI group were higher than those in the IVF group(P＜0.05),but the M Ⅱ rate,poly-spermy rate,high-quality embryo rate,blastocyst formation rate,and available blastocyst rate were significantly lower than those in the IVF group(P＜0.05).The total fertilization rate and poly-spermy rate of the R-ICSI group were significantly higher than those of the ICSI group(P＜0.05),while the blastocyst formation rate was significantly lower than that of the ICSI group(P＜0.05).The proportion of Day 3 embryo cycles transferred in the ICSI group was significantly higher than that in the IVF group(P＜0.05),but there was no significant difference compared to the R-ICSI group(P＞0.05).There was no significant difference in the average number of embryos transferred and the proportion of cycles with Day 5 embryo transferred among the three groups of patients(P＞0.05).In the comparison of clinical outcomes,there were no significant differences in clinical pregnancy rate,implantation rate,abortion rate,delivery rate,multiple pregnancy rate,ectopic pregnancy rate,live birth rate,sex ratio at birth and birth weight among the three groups(P＞0.05). Conclusions:Short-term fertilization combined with R-ICSI can achieve good laboratory and clinical outcomes,which has important application value for patients with failed or low fertilization rate in conventional IVF.

目的 分析体外受精/卵胞浆内单精子注射-胚胎移植(IVF/ICSI-ET)实验室因素对子代出生性别比(SRB)的影响.方法 回顾性分析2010年1月至2019年12月在我院行IVF/ICSI-ET治疗并成功妊娠分娩患者(共19 844个周期)的SRB情况.辅助生殖技术(ART)的主要实验室因素包括受精方式、移植胚胎类型、移植胚胎时期和移植胚胎数目等,故根据不同的受精方式分为IVF组(n=14 626)和ICSI组(n=4 908);根据不同移植胚胎时期分为卵裂胚组(n=8 778)和囊胚组(n=11066);根据不同移植胚胎类型分为新鲜胚胎组(n=11 533)和解冻胚胎组(n=8 311);根据不同移植胚胎个数分为单胚胎组(n=3 871)和双胚胎组(n=14 483).分别观察以上4个实验室因素对SRB的影响;采用单因素及多因素Logistic回归分析各因素与SRB的相关性;并分析2010～2019年我院总SRB的变化趋势.结果 我院2010年1月至2019年12月行IVF/ICSI-ET共出生新生儿24 661名,其中男婴13 532名(54.9％),女婴11 129名(45.1％),该时期ART的SRB(121.6∶100)高于文献报道的自然受孕SRB(106～110.2∶100).不同实验室因素的分组中,IVF组的SRB(126.2∶100)显著高于ICSI组(107.5∶100)(P＜0.05);囊胚组的SRB(129.2∶100)显著高于卵裂胚组(112.8∶100)(P＜0.05);解冻胚胎组的SRB略高于新鲜胚胎组,但差异无统计学意义(P＞0.05);单胚胎组的SRB(130.0∶100)显著高于双胚胎组(120.5∶100)(P＜0.05).Logistic 回归分析结果显示,受精方式[OR=0.854,95％CI(0.803,0.908)]和移植胚胎时期[OR=1.192,95％CI(1.114,1.276)]是SRB的独立影响因素(P＜0.001).2010-2019年我院总SRB没有明显逐年升高或者降低的趋势;在相同年份中,卵裂胚的SRB相对于囊胚有降低趋势,ICSI的SRB相对于IVF有降低趋势.结论 ART的SRB高于自然受孕,IVF受精的SRB显著高于ICSI,囊胚移植的SRB显著高于卵裂期胚胎移植.

High proportion of human embryos produced by in vitro fertilization (IVF) are aneuploidy or have segmental chromosomal errors. Not only a whole chromosome aneuploidy, but also small errors in a chromosome, such as microdeletion can be detected by current next-generation sequencing (NGS) for preimplantation genetic testing (PGT). The prevalence of aneuploidy in donor egg IVF was significantly different between fertility clinics. In the present study, we examined whether different embryo biopsy procedures affect embryonic aneuploidies in donor egg IVF. We did not find significant differences in the samples with abnormal chromosomes between two biopsy methods. When we further analyzed the samples with abnormal chromosomes, we found that 64.0–80.7% of the abnormalities were whole chromosome aneuploidies while 19.3–36.0% were segmental chromosome abnormalities. High embryo implantation rates were obtained after transferring screened euploid blastocysts. These results indicate that blastocyst biopsy procedures may not significantly affect embryo’s chromosomal status, but PGT by high-resolution NGS revealed that high proportions of human embryos derived from donor eggs are not only aneuploidy, but also segmental chromosome abnormal, and screening of small chromosomal errors by NGS is beneficial to patients who use donated eggs for infertility treatment.

Wnt/β-Catenin signaling pathway plays an important role in maintaining self-renewal and pluripotency of human and mouse embryonic stem cells (ESCs). Activation of Wnt/β-Catenin signaling pathway by glycogen synthase kinase-3 (GSK3) inhibitor, the Wnt signaling agonist, could maintain the pluripotency of human and mouse ESCs in the presence of serum. However, the role of signaling pathway in the derivation of buffalo ESCs remains unclear. In this study, we used GSK3 inhibitors (6-bromoindirubin-3'-oxime [BIO] and CHIR99021) and investigated the effect of Wnt/β-Catenin activation on colony formation, proliferation, self-renewal, and pluripotency of Chinese swamp buffalo (buffalo) embryonic stem cell-like cells (ES-like cells), which were isolated from blastocysts. The results showed that buffalo ES-like cells displayed typical morphological characteristics of pluripotent stem cells: positive for alkaline phosphatase staining, expression of pluripotent markers, including OCT4, SOX2, SSEA-1, SSEA-4, LIN28, CH1, NANOG, and the proliferative markers, PCNA and C-MYC. Furthermore, activation of Wnt/β-Catenin signaling pathway by GSK3 inhibitors could promote colony formation and proliferation of buffalo ES-like cells and maintain their undifferentiated state, and upregulate the expression levels of pluripotent-related genes and proliferation-related genes. These results indicated that Wnt/β-Catenin signaling pathway plays an important role in the derivation and pluripotency of buffalo ES-like cells.

To examine the effect of sodium selenite in different concentration on the body weight,superovulation and embryonic developmental competency in mouse,the 6～7 weeks-age mice were randomly divided into four groups and treated with different concentration of sodium selenite solution,such as 0(control group),0.30 mg/kg(low dose group),1.50 mg/kg(medium dose group),2.25 mg/kg(high dose group),respectively.And then the body weight of mice was measured,as well as the number of embryos derived from superovulation and the in vitro developmental rate of embryo was recorded in each group.The results showed that the change of the mice body weight in high dose group(-0.31 g) was significantly lower than those of other three groups(control: 11.06 g,low: 7.95 g,medium: 8.65 g,P0.05),but there was no remarkably different among the low dose group,medium dose group and control group(P0.05).There was no significant difference in the number of embryos derived from mice superovulation among the four groups(P0.05).Compared with the control group,the blastocyst developmental rate of embryo from superovulation by in vitro culture was significantly decreased in high dose group(17.6% vs 27.6%,P0.05),but it had no significant difference with the low dose group(24.3%) and medium dose group(20.6%)(P0.05).These results indicated that high doses of sodium selenite had toxicity on embryonic developmental ability,and its mechanism should be took an intensive study at the cellular and molecular level.

Embryonic stem(ES)cells are derived from the inner cell mass of early blastocyst, and possess the ability of self-renewal and maintenance pluripotency.Wnt signaling pathway plays important roles in maintaining self-renewal of ES cells.Glycogen synthase kinase-3(GSK-3) inhibitors can active Wnt signaling pathway,then promote the self-renewal and cellto-cell adhesion of ES cells.In this paper,we take a review for the effect and mechanism of GSK-3 inhibitors on maintaining self-renewal and cell-to-cell adhesion of ES cells.