Objective:To explore the different metabolites in the follicular fluids (FFs) of polycystic ovary syndrome (PCOS) patients and non-PCOS patients and their effects on the maturation of mouse oocytes and the developmental potential of in vitro fertilization (IVF) embryos. Methods:The clinical data were collected for the retrospective cohort study. Animal experiments were conducted in a randomized controlled trial. This study included PCOS ( n=71) and non-PCOS ( n=70) patients who underwent the first IVF or intracytoplasmic sperm injection (ICSI) cycle in Shanghai JIAI Genetics & IVF institute from June 2019 to June 2020. The patients' FFs were collected and the clinical data from these patients were analyzed. Raman spectroscopy analysis technology was used to detect differences in the metabolic spectra of FFs between the two groups. Mouse GV phase oocytes were placed in FFs from PCOS patients and non-PCOS patients for in vitro maturation (IVM) culture respectively, then the matured mouse oocytes were collected for IVF. The effects of differential metabolites in FFs on mouse oocyte maturation and embryonic development were further explored. The Raman spectrum was also applied to identify the differences of the IVM spent culture media. Results:The MⅡ rate [82.19% (886/1 078)] and day 3 available embryo rate [51.30% (553/1 078)] from PCOS group were significantly lower than those of the non-PCOS group [85.85% (625/728), P=0.038; 53.30% (388/728), P=0.042]. However, there were no significant differences between the two groups in the cumulative clinical pregnancy rate and the cumulative live birth rate (all P>0.05). Raman was capable of distinguishing PCOS from non-PCOS FFs. The characteristic Raman displacement difference between the two groups is mainly concentrated in the 600-1 000 cm -1, as well as 1 168 cm -1, 1 344 cm -1, 1 440 cm -1, 1 504 cm -1, 1 632 cm -1 and 1 664 cm -1. The Raman characteristic shift database showed that the different metabolites of the two sets of FFs samples were mainly concentrated in protein, lipids, free nucleic acis, glucose, cholesterol, carotenoids, and amino acids. Mouse oocyte IVM results showed that the PCOS-FF group had a lower MⅡ rate [49.04% (77/157)] than that of non-PCOS group [65.07% (95/146), P=0.005). IVF results showed the PCOS-FF group had a significantly lower cleavage rate [46.75% (36/77)] than that of non-PCOS group [63.16% (60/95), P=0.031], but there was no significant difference in the blastocyst rate between the two groups ( P>0.05). Conclusion:Differential metabolites detected by Raman spectrum in the PCOS FFs may cause defected maturation of the oocytes, leading to infertility, and Raman spectroscopy is an effective approach towards PCOS diagnosis and the identification of metabolomics differences.

Objective:To explore the different metabolites in the follicular fluids (FFs) of polycystic ovary syndrome (PCOS) patients and non-PCOS patients and their effects on the maturation of mouse oocytes and the developmental potential of in vitro fertilization (IVF) embryos. Methods:The clinical data were collected for the retrospective cohort study. Animal experiments were conducted in a randomized controlled trial. This study included PCOS ( n=71) and non-PCOS ( n=70) patients who underwent the first IVF or intracytoplasmic sperm injection (ICSI) cycle in Shanghai JIAI Genetics & IVF institute from June 2019 to June 2020. The patients' FFs were collected and the clinical data from these patients were analyzed. Raman spectroscopy analysis technology was used to detect differences in the metabolic spectra of FFs between the two groups. Mouse GV phase oocytes were placed in FFs from PCOS patients and non-PCOS patients for in vitro maturation (IVM) culture respectively, then the matured mouse oocytes were collected for IVF. The effects of differential metabolites in FFs on mouse oocyte maturation and embryonic development were further explored. The Raman spectrum was also applied to identify the differences of the IVM spent culture media. Results:The MⅡ rate [82.19% (886/1 078)] and day 3 available embryo rate [51.30% (553/1 078)] from PCOS group were significantly lower than those of the non-PCOS group [85.85% (625/728), P=0.038; 53.30% (388/728), P=0.042]. However, there were no significant differences between the two groups in the cumulative clinical pregnancy rate and the cumulative live birth rate (all P>0.05). Raman was capable of distinguishing PCOS from non-PCOS FFs. The characteristic Raman displacement difference between the two groups is mainly concentrated in the 600-1 000 cm -1, as well as 1 168 cm -1, 1 344 cm -1, 1 440 cm -1, 1 504 cm -1, 1 632 cm -1 and 1 664 cm -1. The Raman characteristic shift database showed that the different metabolites of the two sets of FFs samples were mainly concentrated in protein, lipids, free nucleic acis, glucose, cholesterol, carotenoids, and amino acids. Mouse oocyte IVM results showed that the PCOS-FF group had a lower MⅡ rate [49.04% (77/157)] than that of non-PCOS group [65.07% (95/146), P=0.005). IVF results showed the PCOS-FF group had a significantly lower cleavage rate [46.75% (36/77)] than that of non-PCOS group [63.16% (60/95), P=0.031], but there was no significant difference in the blastocyst rate between the two groups ( P>0.05). Conclusion:Differential metabolites detected by Raman spectrum in the PCOS FFs may cause defected maturation of the oocytes, leading to infertility, and Raman spectroscopy is an effective approach towards PCOS diagnosis and the identification of metabolomics differences.

Objective:To investigate the impact of post-cesarean scar defects and scar diverticulum on the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET). Methods:A total of 523 cases, who had a previous delivery, underwent frozen-thawed single blastocyst transfer cycles during January 1 st, 2014 to January 31 st, 2018 in Shanghai Ji Ai Genetics and IVF Institute. Of which, 256 patients who had undergone cesarean section （C-S） surgery prior, along with 267 patients with a history of one vaginal delivery as control group, were admitted into this study. In addition, C-S group was further divided into 3 subgroups according to the different healing conditions of the uterine incision detected by ultrasound, namely, no severe defect subgroup, scar defects subgroup and diverticulum subgroup. The impact of patients with or without C-S history, patients with or without scar defects and diverticulum on the clinical outcomes of FET cycle was analyzed. Results:There was no difference in age, body mass index (BMI) and thickness of endometrium at the day of progesterone conversion between patients with one vaginal delivery (VD group) and patients with C-S history (C-S group). C-S group showed a significant decrease in implantation rate (48.05%), clinical pregnancy rate (46.48%) and live birth rate (38.28%) compared with VD group (58.05%, P=0.023; 56.93%, P=0.014; 51.31%, P=0.022). There was a significant rise in preterm delivery rate （13.26% vs. 1.46%, P<0.001）, and average gestational age at the time of delivery was decreased [(38.12±1.76) weeks vs. (39.02±1.37) weeks, P<0.001], and neonatal birth weight was also significantly lower in C-S group [(3 372.14±374.25) g vs. (3 438.82±337.30) g, P=0.023]. Further analysis was done to address whether C-S severe defects with diverticulum had an impact on the clinical outcomes. Compared with the vaginal delivery group, the implantation rate, the clinical pregnancy rate, the live birth rate, and the gestational age at the time of delivery in severe scar defects subgroup were significantly reduced ( P=0.021, P=0.026, P=0.005 and P=0.009); and in the scar diverticulum subgroup they were more affected ( P=0.004, P=0.003, P=0.004 and P<0.001), the differences were statistically significant. Conclusion:A history of C-S, particularly a poor healing of incision with scar defects and diverticulum, has a significantly negative impact on the clinical outcomes of IVF-ET as well as a negative impact on gestational age at delivery, and increases the risk of premature birth and lower birth weight newborns.

Objective:To investigate the impact of post-cesarean scar defects and scar diverticulum on the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET). Methods:A total of 523 cases, who had a previous delivery, underwent frozen-thawed single blastocyst transfer cycles during January 1 st, 2014 to January 31 st, 2018 in Shanghai Ji Ai Genetics and IVF Institute. Of which, 256 patients who had undergone cesarean section （C-S） surgery prior, along with 267 patients with a history of one vaginal delivery as control group, were admitted into this study. In addition, C-S group was further divided into 3 subgroups according to the different healing conditions of the uterine incision detected by ultrasound, namely, no severe defect subgroup, scar defects subgroup and diverticulum subgroup. The impact of patients with or without C-S history, patients with or without scar defects and diverticulum on the clinical outcomes of FET cycle was analyzed. Results:There was no difference in age, body mass index (BMI) and thickness of endometrium at the day of progesterone conversion between patients with one vaginal delivery (VD group) and patients with C-S history (C-S group). C-S group showed a significant decrease in implantation rate (48.05%), clinical pregnancy rate (46.48%) and live birth rate (38.28%) compared with VD group (58.05%, P=0.023; 56.93%, P=0.014; 51.31%, P=0.022). There was a significant rise in preterm delivery rate （13.26% vs. 1.46%, P<0.001）, and average gestational age at the time of delivery was decreased [(38.12±1.76) weeks vs. (39.02±1.37) weeks, P<0.001], and neonatal birth weight was also significantly lower in C-S group [(3 372.14±374.25) g vs. (3 438.82±337.30) g, P=0.023]. Further analysis was done to address whether C-S severe defects with diverticulum had an impact on the clinical outcomes. Compared with the vaginal delivery group, the implantation rate, the clinical pregnancy rate, the live birth rate, and the gestational age at the time of delivery in severe scar defects subgroup were significantly reduced ( P=0.021, P=0.026, P=0.005 and P=0.009); and in the scar diverticulum subgroup they were more affected ( P=0.004, P=0.003, P=0.004 and P<0.001), the differences were statistically significant. Conclusion:A history of C-S, particularly a poor healing of incision with scar defects and diverticulum, has a significantly negative impact on the clinical outcomes of IVF-ET as well as a negative impact on gestational age at delivery, and increases the risk of premature birth and lower birth weight newborns.

Objective To study the relationship between reactive oxygen species (ROS) as well as total antioxidant capacity ( TAC ) within follicle fluid and body mass index ( BMI ) in patients with polycystic ovary syndrome (PCOS).Methods All patients enrolled in this study were infertile women receiving in vitro fertilization-embryo transfer (IVF-ET) treatment.55 PCOS patients were divided into over-weight group ( n =23 ) and non-over-weight group ( n =32 ).Another 55 age-matched non-PCOS women were also divided into control group ( n =30) and overweight group ( n =25 ).Plasma sex hormone,triglycerides,and total cholesterol were determined.On oocyte retrieval day after ovarian stimulation,ROS and TAC in follicular fluid were assayed.Results Subjects in over-weight and PCOS over-weight groups had higher triglycerides than those in control and PCOS non-over-weight groups [ ( 1.9 ±1.1,1.7 ± 0.9,1.0 ± 0.5,1.2 ± 0.7 ) mmol/L,respectively,all P＜0.05],so did total cholesterol [ ( 4.8 ± 1.2,5.2 ± 1.1,4.0 ± 0.6) mmol/L,respectively,all P＜0.05].In PCOS over-weight group,ROS and ROS/TAC within follicular fluid were ( 35.4 ± 6.7 ) RLU/S and 39.8 ± 22.0,both were higher than those in the other 3 groups ( all P＜0.05).TAC [ (0.8 ± 0.5 ) Mm] was lower in PCOS over-weight group than that among the other 3 groups( all P＜0.05 ).ROS/TAC was higher in PCOS non-over-weight group than that of control group ( 26.5 ± 14.5 vs 14.2 ± 12.5,P＜0.05 ).Univariate analysis showed that both ROS and ROS/TAC within follicular fluid in PCOS patients were positively correlated with BMI ( r =0.34 and r =0.32,both P＜0.05 ).Conclusion Abnormal oxidative stress exists in follicular fluid of PCOS patients,and the oxidative stress parameters show positive correlation with BMI.

The hypoxanthine-guanine phosphoribosyl transferase (HGPRT) gene mutation is responsible for gouty arthritis, kidney stone, and Lesch-Nyhan Syndrome (LNS). It has been reported that the expression of HGPRT is decreased or even absent in these diseases. Rabbits are an ideal model for studying the pathology of these diseases. Therefore, the development of an HGPRT-knockdown rabbit model will be highly beneficial m such studies. Stable HGPRT-knoekdown transgenie fibroblast lines were generated by transfecting rabbit fibroblasts with RNA interference (RNAi) plasmids. Polymerase chain reaction (PCR) analyses indicated that the average positive rate was 83.3%. The mRNA and protein levels of HGPRT in the transgenic fibroblast lines were significantly lower than that in the control. Transgenic rabbit blastocysts were derived after performing nuclear transfer. The results show that RNAi can be used to stably knock down expression of the HGPRT in rabbit fibroblasts and further improvements in related technologies will facilitate the use of this method for the generation of HGPRT-knockdown rabbits.