Objective:To explore the mechanism of peripheral blood mononuclear cells (PBMCs)-derived exosomes on improving endometrial receptivity by miR-1306.Methods:The models of mice with implantation disorders were prepared by subcutaneous injection of mifepristone. And they were divided into implantation disorder group and PBMCs intervention group, 12 mice in each group. The normal group was set up. PBMCs intervention group was given intrauterine injection of PBMCs-derived exosomes, while normal group and implantation disorder group were given the same volume of buffer solution. The pregnancy rates and number of embryos implantation in different groups were compared. The expression of miR-1306 in endometrial tissues was detected by RT-PCR. The levels of reactive oxygen species (ROS), interleukin-6 (IL-6), monocyte chemcattractant protein-1 (MCP-1) and superoxide dismutase (SOD) in endometrial tissues were detected by enzyme-linked immunosorbent assay. The expressions of pregnancy-related proteins in endometrial tissues were detected by Western blotting. The endometrial epithelial cells were divided into control group, experimental group, negative control group and miR-1306 inhibitor group. Except control group, cells were co-cultured with PBMCs-derived exosomes by Transwell in the other groups. The activity, proliferation and apoptosis of cells were detected by MTT, Edu and Annexin-V/PI flow, respectively. The level of ROS in cells was detected by kits. And expressions of the related proteins were detected by Western blotting.Results:The pregnancy rate [8.33% (1/12), number of embryos implantation [0(0, 0)], expression of miR-1306 in endometrial tissue (0.24±0.05) and SOD level [(5.66±0.72) U/mL] in implantation disorder group were significantly lower than those in normal group [100% (12/12), 16.50 (14.00, 19.00), 1.03±0.05, (8.69±1.21) U/mL, P<0.05], while levels of ROS [(4.87±0.39) U/mL], IL-6 [(116.51±5.78) ng/L] and MCP-1 in endometrial tissues [(36.84±3.56) μg/L], and expressions of KIR2DL4 (0.87±0.06), nuclear factor erythroid-2-related factor 2 (Nrf2, 0.76±0.06), Kelch-like ECH-associated protein1 (Keap1, 0.79±0.05), receptor interacting protein 1 (RIP1, 0.94±0.04) and RIP3 (0.86±0.05) were significantly higher than those in normal group [(2.41±0.19) U/mL, (83.79±6.68) ng/L, (12.32±2.09) ng/mL, 0.27±0.03, 0.31±0.05, 0.23±0.04, 0.34±0.03, 0.31±0.05, all P<0.05]. The pregnancy rate [75.00% (9/12)], number of embryos implantation [13.00 (13.00, 14.75)], expression of miR-1306 in endometrial tissues (0.82±0.05) and SOD level [(7.24±0.84) U/mL] in PBMCs intervention group were significantly higher than those in implantation disorder group (all P<0.05), while levels of ROS [(3.43±0.30) U/mL], IL-6 [(94.69±3.99) ng/L] and MCP-1 in endometrial tissues [(27.03±3.48) μg/L], and expressions of KIR2DL4 (0.54±0.08), Nrf2 (0.48±0.05), Keap1 (0.43±0.05), RIP1 (0.56±0.05) and RIP3 (0.49±0.03) were significantly lower than those in implantation disorder group (all P<0.05). The cells activity [(126.63±1.25)%], proliferation rate [(53.54±2.82)%] and ROS level [(3.12±0.31) U/mL] in experimental group were significantly higher than those in control group [100%, (23.18±3.07)%, (2.51±0.28) U/mL, all P<0.05], while apoptosis rate [(5.69±0.47)%], expressions of KIR2DL4 (0.36±0.06), Nrf2 (0.30±0.06), Keap1 (0.26±0.04), RIP1 (0.27±0.05) and RIP3 (0.26±0.06) were lower than those in control group [(27.13±2.97)%, 0.84±0.06, 0.75±0.05, 0.68±0.05, 0.80±0.06, 0.80±0.07, all P<0.05]. The cells activity [(83.48±5.34)%] and proliferation rate [(38.42±4.28)%] in miR-1306 inhibitor group were significantly lower than those in negative control group [(127.12±4.08)%, (53.57±2.09)%, all P<0.05], while apoptosis rate [(13.63±1.77)%], ROS level [(6.49±0.62) U/mL], expressions of KIR2DL4 (0.67±0.07), Nrf2 (0.57±0.05), Keap1 (0.50±0.05), RIP1 (0.64±0.06) and RIP3 (0.61±0.08) were significantly higher than those in negative control group [(5.71±0.78)%, (3.23±0.31) U/mL, 0.36±0.07, 0.30±0.07, 0.27±0.06, 0.28±0.07, 0.28±0.06, all P<0.05]. Conclusion:PBMCs-derived exosomes can improve pregnancy rate in mice with implantation disorders, which may improve inflammatory response by promoting miR-1306 expression and inhibiting KIR2DL4 expression, and is also related to relieving excessive oxidative stress and apoptosis of endometrium.

Objective:To explore the mechanism of peripheral blood mononuclear cells (PBMCs)-derived exosomes on improving endometrial receptivity by miR-1306.Methods:The models of mice with implantation disorders were prepared by subcutaneous injection of mifepristone. And they were divided into implantation disorder group and PBMCs intervention group, 12 mice in each group. The normal group was set up. PBMCs intervention group was given intrauterine injection of PBMCs-derived exosomes, while normal group and implantation disorder group were given the same volume of buffer solution. The pregnancy rates and number of embryos implantation in different groups were compared. The expression of miR-1306 in endometrial tissues was detected by RT-PCR. The levels of reactive oxygen species (ROS), interleukin-6 (IL-6), monocyte chemcattractant protein-1 (MCP-1) and superoxide dismutase (SOD) in endometrial tissues were detected by enzyme-linked immunosorbent assay. The expressions of pregnancy-related proteins in endometrial tissues were detected by Western blotting. The endometrial epithelial cells were divided into control group, experimental group, negative control group and miR-1306 inhibitor group. Except control group, cells were co-cultured with PBMCs-derived exosomes by Transwell in the other groups. The activity, proliferation and apoptosis of cells were detected by MTT, Edu and Annexin-V/PI flow, respectively. The level of ROS in cells was detected by kits. And expressions of the related proteins were detected by Western blotting.Results:The pregnancy rate [8.33% (1/12), number of embryos implantation [0(0, 0)], expression of miR-1306 in endometrial tissue (0.24±0.05) and SOD level [(5.66±0.72) U/mL] in implantation disorder group were significantly lower than those in normal group [100% (12/12), 16.50 (14.00, 19.00), 1.03±0.05, (8.69±1.21) U/mL, P<0.05], while levels of ROS [(4.87±0.39) U/mL], IL-6 [(116.51±5.78) ng/L] and MCP-1 in endometrial tissues [(36.84±3.56) μg/L], and expressions of KIR2DL4 (0.87±0.06), nuclear factor erythroid-2-related factor 2 (Nrf2, 0.76±0.06), Kelch-like ECH-associated protein1 (Keap1, 0.79±0.05), receptor interacting protein 1 (RIP1, 0.94±0.04) and RIP3 (0.86±0.05) were significantly higher than those in normal group [(2.41±0.19) U/mL, (83.79±6.68) ng/L, (12.32±2.09) ng/mL, 0.27±0.03, 0.31±0.05, 0.23±0.04, 0.34±0.03, 0.31±0.05, all P<0.05]. The pregnancy rate [75.00% (9/12)], number of embryos implantation [13.00 (13.00, 14.75)], expression of miR-1306 in endometrial tissues (0.82±0.05) and SOD level [(7.24±0.84) U/mL] in PBMCs intervention group were significantly higher than those in implantation disorder group (all P<0.05), while levels of ROS [(3.43±0.30) U/mL], IL-6 [(94.69±3.99) ng/L] and MCP-1 in endometrial tissues [(27.03±3.48) μg/L], and expressions of KIR2DL4 (0.54±0.08), Nrf2 (0.48±0.05), Keap1 (0.43±0.05), RIP1 (0.56±0.05) and RIP3 (0.49±0.03) were significantly lower than those in implantation disorder group (all P<0.05). The cells activity [(126.63±1.25)%], proliferation rate [(53.54±2.82)%] and ROS level [(3.12±0.31) U/mL] in experimental group were significantly higher than those in control group [100%, (23.18±3.07)%, (2.51±0.28) U/mL, all P<0.05], while apoptosis rate [(5.69±0.47)%], expressions of KIR2DL4 (0.36±0.06), Nrf2 (0.30±0.06), Keap1 (0.26±0.04), RIP1 (0.27±0.05) and RIP3 (0.26±0.06) were lower than those in control group [(27.13±2.97)%, 0.84±0.06, 0.75±0.05, 0.68±0.05, 0.80±0.06, 0.80±0.07, all P<0.05]. The cells activity [(83.48±5.34)%] and proliferation rate [(38.42±4.28)%] in miR-1306 inhibitor group were significantly lower than those in negative control group [(127.12±4.08)%, (53.57±2.09)%, all P<0.05], while apoptosis rate [(13.63±1.77)%], ROS level [(6.49±0.62) U/mL], expressions of KIR2DL4 (0.67±0.07), Nrf2 (0.57±0.05), Keap1 (0.50±0.05), RIP1 (0.64±0.06) and RIP3 (0.61±0.08) were significantly higher than those in negative control group [(5.71±0.78)%, (3.23±0.31) U/mL, 0.36±0.07, 0.30±0.07, 0.27±0.06, 0.28±0.07, 0.28±0.06, all P<0.05]. Conclusion:PBMCs-derived exosomes can improve pregnancy rate in mice with implantation disorders, which may improve inflammatory response by promoting miR-1306 expression and inhibiting KIR2DL4 expression, and is also related to relieving excessive oxidative stress and apoptosis of endometrium.

Objective　To explore the correlations between serum hypoxia inducible factor-1α （HIF-1α），aquaporin-9 （AQP9） and hemorrhagic transformation （HT） in patients with acute ischemic stroke （AIS） and its clinical significance. Methods　183 newly diagnosed patients with acute AIS from October 2017 to April 2019 were divided into HT group （n=84） and non HT group （n=99），the baseline data of gender，age，body mass index （BMI），proportions of hypertension，diabetes，hyperlipidemia，the baseline NIHSS score，baseline systolic blood pressure，baseline diastolic blood pressure，baseline blood glucose，thrombolytic time window and serum samples were collected. The expressions of HIF-1α and AQP9 proteins were detected by Western blot （WB）；Pearson’s method was used to analyze the correlation between HIF-1α and AQP9 levels in patients with AIS in HT group；the risk factors of HT in AIS patients were analyzed by logistic regression；Using NIHSS score，thrombolysis time window，serum HIF-1α，AQP9 levels as independent variables，the working characteristic curve （ROC） of subjects was drawn to analyze the predictive value of HT in AIS patients. Results　Compared with the non HT group，the baseline NIHSS score，baseline systolic blood pressure，baseline diastolic blood pressure，baseline blood glucose，thrombolytic time window，serum HIF-1α，AQP9 levels of AIS patients in HT group were significantly higher （P<0.05）. There was a positive correlation between the levels of HIF-1α and AQP9 proteins in patients with AIS in HT group （r=0.679，P<0.05）. NIHSS score，thrombolysis time window and serum HIF-1α，AQP9 protein levels were independent risk factors for HT in AIS patients after thrombolysis （P<0.05）. The AUCs of NIHSS score，thrombolysis time window，serum HIF-1α and AQP9 in the diagnosis of HT in AIS patients were 0.707，0.790，0.881 and 0.869 respectively，the cutoff values were 13.39 points，296.31 min，0.33 and 0.32 respectively，the sensitivities were 44%，66.70%，86.90% and 83.30% respectively，and the specificities were 91.90%，87.90%，87.90% and 88.90% respectively. The AUC of the combination of four methods in the diagnosis of HT in AIS patients was 0.980，the sensitivity and specificity were 94.00% and 94.90%，respectively，and the diagnostic efficiency of the four methods was significantly higher than that of single index. Conclusion　The elevated levels of HIF-1α and AQP9 were closely related to the occurrence of HT in AIS patients，and the combination of NIHSS score and thrombolysis time window could significantly improve the predictive value of HT in AIS，which may have some clinical reference significance.

Objective:To explore the effects of phloroglucinol on transfer outcome of in vitro fertilization-embryo transfer (IVF-ET). Methods:Retrospective analysis of 4289 cases of infertility patients treated with IVF-ET in the Department of Reproductive Medicine, the Second Affiliated Hospital of Zhengzhou University from January 1, 2015 to November 30, 2018 was performed. According to the pregnancy outcome, all patients were divided into pregnancy group and non-pregnancy group to analyze the risk factors affecting the pregnancy outcome. According to medication before transfer, all patients were divided into no medication group (group A), preoperative intravenous infusion of atosiban group (group B), preoperative and intraoperative injection of atosiban group (group C) and interoperative intravenous infusion of phloroglucinol group (group D). The pregnancy outcomes, occurrence of adverse reactions during and after treatment were compared among the four groups.Results:Age, number of high-quality embryos, and medication methods before transfer were independent factors influencing pregnancy outcomes in patients undergoing IVF-ET ( P=0.031, P=0.039, P=0.020). The embryo implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group D were significantly higher than those in group A (all P<0.001), group B (all P<0.001) and group C ( P<0.001， P<0.001， P=0.001), while the ectopic pregnancy rate was significantly lower than that in group A, group B and group C (all P<0.001). The abortion rate in group D was lower than that in group A ( P<0.001). During treatment period and after treatment, there were no discomfort complains and allergic reactions in the four groups. There were no birth defects in all born babies. Conclusion:The medication method is an independent factor influencing pregnancy outcomes of patients undergoing IVF-ET. Intraoperative intravenous infusion of phloroglucinol can significantly increase embryo implantation rate and pregnancy rate of IVF-ET patients, and reduce their abortion rate.

Objective:To explore the effects of phloroglucinol on transfer outcome of in vitro fertilization-embryo transfer (IVF-ET). Methods:Retrospective analysis of 4289 cases of infertility patients treated with IVF-ET in the Department of Reproductive Medicine, the Second Affiliated Hospital of Zhengzhou University from January 1, 2015 to November 30, 2018 was performed. According to the pregnancy outcome, all patients were divided into pregnancy group and non-pregnancy group to analyze the risk factors affecting the pregnancy outcome. According to medication before transfer, all patients were divided into no medication group (group A), preoperative intravenous infusion of atosiban group (group B), preoperative and intraoperative injection of atosiban group (group C) and interoperative intravenous infusion of phloroglucinol group (group D). The pregnancy outcomes, occurrence of adverse reactions during and after treatment were compared among the four groups.Results:Age, number of high-quality embryos, and medication methods before transfer were independent factors influencing pregnancy outcomes in patients undergoing IVF-ET ( P=0.031, P=0.039, P=0.020). The embryo implantation rate, the biochemical pregnancy rate and the clinical pregnancy rate of group D were significantly higher than those in group A (all P<0.001), group B (all P<0.001) and group C ( P<0.001， P<0.001， P=0.001), while the ectopic pregnancy rate was significantly lower than that in group A, group B and group C (all P<0.001). The abortion rate in group D was lower than that in group A ( P<0.001). During treatment period and after treatment, there were no discomfort complains and allergic reactions in the four groups. There were no birth defects in all born babies. Conclusion:The medication method is an independent factor influencing pregnancy outcomes of patients undergoing IVF-ET. Intraoperative intravenous infusion of phloroglucinol can significantly increase embryo implantation rate and pregnancy rate of IVF-ET patients, and reduce their abortion rate.

OBJECTIVETo evaluate the effect of basal serum luteinizing hormone (LH) and luteinizing hormone/follicle-stimulating hormone (LH/FSH) ratio on the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET) in patients with polycystic ovary syndrome (PCOS).

METHODSA retrospective analysis was performed of 134 IVF cycles in patients with PCOS. The cycles were classified into 2 groups according to serum levels of LH and also into 2 groups according to LH/FSH ratio, namely group A1 (LH≤10 IU/L), group A2 (LH>10 IU/L), group B1 (LH/FSH ratio<2), and group B2 (LH/FSH ratio≥2). The clinical characteristics, embryological data and pregnancy outcomes were compared between the groups.

RESULTSPatients in group A2 showed significantly higher FSH, T level, and LH/FSH ratio with a greater number of oocytes retrieved than those in group A1, but the time for down-regulation, duration of stimulation, AFC, LH and LH/FSH on the first day of stimulation, embryological data and pregnancy outcomes did not differ significantly between the two groups. Compared with group B1, group B2 showed higher basal LH, E2 level on the day of HCG, more oocytes retrieved and lower dose of gonadotropins used, but the time for down-regulation, duration of stimulation, LH and LH/FSH on the first day of stimulation and pregnancy outcomes were comparable between the two groups.

CONCLUSIONA high basal LH level or a high LH/FSH ratio does not produce obvious deleterious effect on the clinical outcomes of IVF-ET in women with PCOS who take oral contraceptives for pretreatment before long GnRH-agonist protocol.

Adult ; Female ; Fertilization in Vitro ; methods ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Polycystic Ovary Syndrome ; blood ; Pregnancy ; Pregnancy Rate ; Retrospective Studies