Objective To investigate the regulation of SIRT1 on ovarian granulosa cell in women with poor ovarian response.Methods A total of 60 women who underwent IVF/ICSI-ET were included in this study,30 women in poor ovarian response(POR)group and 30 women in normal ovarian response(NOR)group.The granulosa cells in follicular fluid were isolated and purified by density gradient centrifugation.The granulosa cells of POR group were treated with SIRT1 agonist resveratrol(RESV)or inhibitor EX527,respectively,and the control group was granulosa cells treated without drugs.mRNA levels of SIRT1 and key enzymes of steroid hormone synthesis(StAR,CYP19,CYP17)in granulosa cells were detected by qPCR.The levels of SIRT1,steroid hormone synthesis key enzymes protein and apoptosis-related protein(Bcl-2,Caspase-3)in granulosa cells were detected by Western blot.The apoptosis rate of granulosa cells was detected by TUNEL method.Determination of estradiol in granulosa cell culture medium by chemiluminescence.Results The mRNA and protein levels of SIRT1 in granulosa cells in POR group were lower than those in NOR group(P = 0.003,P<0.001).RESV up-regulated SIRT1 mRNA and protein levels(P<0.001),up-regulated mRNA and protein levels of steroid hormone synthesis key enzyme(P<0.05).EX527 down-regulated SIRT1 mRNA and protein levels(P = 0.023,0.001),down-regulated mRNA and protein levels of steroid hormone synthesis key enzymes(P<0.05).After RESV treatment,the apoptosis rate of granulosa cells was decreased(P<0.001),the Bcl-2 protein level was increased(P<0.001),and the Caspase-3 protein level was decreased(P<0.001).After EX527 treatment,the apoptosis rate of granulosa cells was increased(P<0.001),the Bcl-2 protein level was decreased(P = 0.003),and the Caspase-3 protein level was increased(P<0.001).Conclusion The SIRT1 level in granulosa cells of POR women was lower than that of NOR women.RESV up-regulated SIRT1 expression in granulosa cells.SIRT1 inhibited granulosa cell apoptosis,increased the level of steroid hormone synthesis key enzyme,and promoted estradiol synthesis in granulosa cell.Therefore,SIRT1 activators such as RESV may be a therapeutic agent to improve ovarian reactivity and increase the number of oocytes obtained in women with POR.

Objective:To investigate whether the cyclosporin A (CsA) can improve the blastocyst implantation with mild impaired of trophoblast cells of mice.Methods:1) Totally 30 ICR mice were intraperitoneally injected with 5 mg/kg CsA, then the blood concentrations of CsA in mice were detected at 1 h, 3 h, 4 h, 6 h, 8 h, 10 h, 13 h, 14 h, 16 h and 20 h after administration using chemiluminescence method. 2) A total of 36 mice were randomly divided into control group and experimental group (CsA group); the experimental group was intraperitoneally injected with 5 mg/kg CsA and control group was injected with the equivalent dose olive oil of the same body weight. Embryos were classified into A, B, C types according to the guidelines of Gardner and experimental needs. Embryos were transferred to the uterus in the two groups. The embryo implantation rate was calculated and the leukemia inhibitory factor (LIF) mRNA level was measured by real time fluorescence quantitative PCR in 5.5 d postcoitum (dpc).Results:1) The blood concentration of CsA in mice reached a peak at 6-10 h after administration. 2) The embryo implantation rate of B type [73.9% (34/46)] had significantly improved in experimental group [50.0% (23/46), P=0.018]. 3) The expression of LIF mRNA level was not significantly different between the two groups ( P>0.05). Conclusion:When CsA was administered at a dose of 5 mg/kg, CsA can improve the embryo implantation rate with mild impaired trophoblast cells. CsA might become a potential drug to increase the success rates of the in vitro fertilization.

Objective:To investigate whether the cyclosporin A (CsA) can improve the blastocyst implantation with mild impaired of trophoblast cells of mice.Methods:1) Totally 30 ICR mice were intraperitoneally injected with 5 mg/kg CsA, then the blood concentrations of CsA in mice were detected at 1 h, 3 h, 4 h, 6 h, 8 h, 10 h, 13 h, 14 h, 16 h and 20 h after administration using chemiluminescence method. 2) A total of 36 mice were randomly divided into control group and experimental group (CsA group); the experimental group was intraperitoneally injected with 5 mg/kg CsA and control group was injected with the equivalent dose olive oil of the same body weight. Embryos were classified into A, B, C types according to the guidelines of Gardner and experimental needs. Embryos were transferred to the uterus in the two groups. The embryo implantation rate was calculated and the leukemia inhibitory factor (LIF) mRNA level was measured by real time fluorescence quantitative PCR in 5.5 d postcoitum (dpc).Results:1) The blood concentration of CsA in mice reached a peak at 6-10 h after administration. 2) The embryo implantation rate of B type [73.9% (34/46)] had significantly improved in experimental group [50.0% (23/46), P=0.018]. 3) The expression of LIF mRNA level was not significantly different between the two groups ( P>0.05). Conclusion:When CsA was administered at a dose of 5 mg/kg, CsA can improve the embryo implantation rate with mild impaired trophoblast cells. CsA might become a potential drug to increase the success rates of the in vitro fertilization.

Development of thoracolumbar vertebra (TLV) and rib primordium (RP) is a common evolutionary feature across vertebrates, although whole-organism analysis of the expression dynamics of TLV- and RP-related genes has been lacking. Here, we investigated the single-cell transcriptome landscape of thoracic vertebra (TV), lumbar vertebra (LV), and RP cells from a pig embryo at 27 days post-fertilization (dpf) and identified six cell types with distinct gene expression signatures. In-depth dissection of the gene expression dynamics and RNA velocity revealed a coupled process of osteogenesis and angiogenesis during TLV and RP development. Further analysis of cell type-specific and strand-specific expression uncovered the extremely high level of HOXA10 3'-UTR sequence specific to osteoblasts of LV cells, which may function as anti-HOXA10-antisense by counteracting the HOXA10-antisense effect to determine TLV transition. Thus, this work provides a valuable resource for understanding embryonic osteogenesis and angiogenesis underlying vertebrate TLV and RP development at the cell type-specific resolution, which serves as a comprehensive view on the transcriptional profile of animal embryo development.

Objective To investigate the influence of total progressively motile sperm count(TPMSC) after treatment on clinical outcomes of intrauterine insemination(IUI) with the husband′s sperm in ovulation-promoting cycles.Methods The clinical data in 4179 cases undergoing IUI with the husband′s sperm in ovulation-promoting cycles were retrospectively analyzed.The correlation between clinical pregnancy rate and TPMSC was analyzed.Results Among all the clinical data,TPMSC was to 100×106 in occasional live sperm.TPMSC<0.15×106 was in 15 cases,1 case had pregnancy (live sperm was occasionally seen on IUI day after sperm processing).Ten cases of TPMSC >60×106 had no pregnancy.A total of 4 154 cases of TPMSC (0.15-60.00)×106 were analyzed.The female age,duration of infertility,number of follicles and endometrial thickness(EDM) had no statistical differences among various groups.The clinical pregnancy rate was 13.5%(576/4 154),the group with the highest clinical pregnancy rate was (5.00-<10.00)×106.But there was no statistically significant difference in clinical pregnancy rate among groups(P=0.133).Conclusion Performing IUI in PMSC (0.15-60.00)×106 after processing can get preferable pregnancy rates.