Objective:To explore the effect of vaginal microbiota disorder on pregnancy outcomes in the first-time frozen-thawed embryo transfer (FET) patients and perinatal outcomes in single pregnancy live delivery patients.Methods:The clinical data of 2 299 cycles of FET patients in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from January 2021 to December 2022 were collected in a retrospective cohort study. According to the types of dominant bacteria in the vaginal microbiota before transplantation, they were divided into three groups: control group (dominant bacteria were Lactobacillus, which was Gram positive bacteria, 1 849 cycles), the Gram negative bacilli group (191 cycles), and the Gram positive cocci group (259 cycles). Baseline data and pregnancy outcomes were compared among the three groups. The perinatal outcomes of select single pregnancy live birth FET patients who met the inclusion criteria were further analyzed among the three groups. The main observation indicators were live birth rate, miscarriage rate, and preterm birth rate. A multivariate logistic regression model was used to control confounding factors in the main observation indicators, and to analyze the relationship between dominant bacterial types in the vaginal microbiota and live birth rate, miscarriage rate, and preterm birth rate.Results:The difference in endometrial thickness on the day of transplantation among control group, Gram negative bacilli group, and Gram positive cocci group was statistically significant [(9.38±1.58) mm, (9.56±1.70) mm, and (9.84±1.74) mm, respectively, P<0.001]. In the endometrium preparation methods, the proportion of down-regulation+artificial cycle patients in the Gram positive cocci group was higher than that in control group, and the difference was statistically significant [13.13% (34/259) and 7.46% (138/1 849), respectively, P<0.016 7]. In the pregnancy outcomes, there was a statistically significant difference in live birth rate among control group, Gram negative bacilli group, and Gram positive cocci group [49.86% (922/1 849), 49.21% (94/191) and 41.70% (108/259), respectively, P=0.048]. The live birth rate of the Gram positive cocci group was lower than that of control group, and the difference between the two groups was statistically significant ( P<0.016 7). There were no statistically significant differences in other pregnancy outcomes and perinatal outcomes of single pregnancy live birth FET patients (all P>0.05). The multivariate logistic regression model corrected for female age, infertility years, basal follicle stimulating hormone, anti-Müllerian hormone, proportion of single embryo transfer, proportion of single blastocyst transfer, endometrial thickness on transfer day, and endometrial preparation methods, Gram positive cocci were independent risk factors for live brith after FET transplantation (a OR=0.73, 95% CI: 0.55-0.95, P=0.021). Conclusion:The dominant bacteria in the vaginal microbiota before embryo transfer are Gram positive cocci, which may be related to a decrease in live birth rate in first-time FET patients, but not significantly related to the perinatal outcomes.

Objective:To explore the effect of vaginal microbiota disorder on pregnancy outcomes in the first-time frozen-thawed embryo transfer (FET) patients and perinatal outcomes in single pregnancy live delivery patients.Methods:The clinical data of 2 299 cycles of FET patients in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from January 2021 to December 2022 were collected in a retrospective cohort study. According to the types of dominant bacteria in the vaginal microbiota before transplantation, they were divided into three groups: control group (dominant bacteria were Lactobacillus, which was Gram positive bacteria, 1 849 cycles), the Gram negative bacilli group (191 cycles), and the Gram positive cocci group (259 cycles). Baseline data and pregnancy outcomes were compared among the three groups. The perinatal outcomes of select single pregnancy live birth FET patients who met the inclusion criteria were further analyzed among the three groups. The main observation indicators were live birth rate, miscarriage rate, and preterm birth rate. A multivariate logistic regression model was used to control confounding factors in the main observation indicators, and to analyze the relationship between dominant bacterial types in the vaginal microbiota and live birth rate, miscarriage rate, and preterm birth rate.Results:The difference in endometrial thickness on the day of transplantation among control group, Gram negative bacilli group, and Gram positive cocci group was statistically significant [(9.38±1.58) mm, (9.56±1.70) mm, and (9.84±1.74) mm, respectively, P<0.001]. In the endometrium preparation methods, the proportion of down-regulation+artificial cycle patients in the Gram positive cocci group was higher than that in control group, and the difference was statistically significant [13.13% (34/259) and 7.46% (138/1 849), respectively, P<0.016 7]. In the pregnancy outcomes, there was a statistically significant difference in live birth rate among control group, Gram negative bacilli group, and Gram positive cocci group [49.86% (922/1 849), 49.21% (94/191) and 41.70% (108/259), respectively, P=0.048]. The live birth rate of the Gram positive cocci group was lower than that of control group, and the difference between the two groups was statistically significant ( P<0.016 7). There were no statistically significant differences in other pregnancy outcomes and perinatal outcomes of single pregnancy live birth FET patients (all P>0.05). The multivariate logistic regression model corrected for female age, infertility years, basal follicle stimulating hormone, anti-Müllerian hormone, proportion of single embryo transfer, proportion of single blastocyst transfer, endometrial thickness on transfer day, and endometrial preparation methods, Gram positive cocci were independent risk factors for live brith after FET transplantation (a OR=0.73, 95% CI: 0.55-0.95, P=0.021). Conclusion:The dominant bacteria in the vaginal microbiota before embryo transfer are Gram positive cocci, which may be related to a decrease in live birth rate in first-time FET patients, but not significantly related to the perinatal outcomes.

Objective: To establish the culture technique for culturing γδ T cells in vitro and evaluate the basic characteristics, security and anti-tumor effect of the cultured γδ T cells. Methods: Phytohemagglutinin, zoledronic acid, interleukin-2 and interleukin -7 were used to induce the abundant expansion of peripheral blood mononuclear cells in vitro. Flow cytometry assay, in vitro killing assay and mouse model of human lung cancer were also adopted to assess the characteristics and the anti-tumor effect of cultured γδ T cells. Additionally, the contamination of exogenous agents and the acute toxicity of γδ T cells were determined. Results: After culturing 14-16 days in vitro, the total number of γδ T cells was more than 1.0×10¹⁰. Among these γδ T cells, CD3⁺ γδ TCR⁺ cells accounted for more than 90%. None of contaminations of bacteria, fungi, mycoplasma and virus were observed. At effect target ratio (E/T ratio) of 50/1, killing efficiency of γδ T cells cultured in vitro to SK-MES-1, Ho8910, A549 and K562 reached more than 65%. In vivo experiments showed that the tumor volume of γδ T-treated mice was (828.99±61.05) mm^3, significantly lower than (1 723.51±84.30) mm³ of the control mice (P<0.05). Meanwhile, no acute toxicity effect was observed in γδ T cells treated mice. Conclusion The number, purity and activity of γδT cells cultured in our institute can reach the requirement of clinical application, and the γδT cells also display strong cytotoxic activity against tumor cells such as lung cancer, ovarian cancer and leukemia.

Objective To discuss the relationships between regular pattern changes of plasma fibrinogen (Fib),D-dimer and fibrinogen degradation products (FDP) levels and the recent dissolution of thrombus in patients with pulmonary embolism (PE) in 14 days after treatment.Methods A prospective study was conducted.PE patients admitted to Departments of Respiratory Disease in 4 hospitals from January 2015 to March 2016 were enrolled and all of them were treated with thrombolysis and/or anticoagulation after admission.The computed tomographic pulmonary angiography (CTPA) was examined pre-treatment and 14 days post-treatment in PE patients.The pulmonary artery obstruction index (PAOI) was assessed according to the Mastora scoring method to estimate the thrombus load.The plasma Fib,D-dimer and FDP levels were measured before and on 1,2,3,5,7 and 14 days after treatment,and the relationships between the change regularities of these parameters and PAOI were also analyzed.Results A total of 42 PE patients were enrolled.The curve change of coagulation-fibrinolytic system parameters in 14 days after treatment showed that the Fib level was raised to its peak on the 3rd day after treatment and then decreased (g/L:4.24 ± 1.45 vs.3.83 ± 1.56),representing that its curve change was in accordance with the quadratic model (P =0.095).After treatment,the D-dimer and FDP levels were kept declining,they were reached the valley on 14th day [D-dimer (mg/L):1.58 ± 1.38vs.8.84 ± 6.35,FDP (mg/L):4.23 ± 3.63 vs.23.41 ± 16.54],and their curve changes were in accordance with the cubic model (F was 32.190 and 34.326,respectively both P =0.000).The PAOI variation before and 14 days after treatment [(18.77 ± 14.22)％] was not correlated with Fib variation [(1.20 ± 0.93) g/L,r =-0.194,P =0.219],but was positively correlated with D-dimer variation [(7.29 ± 7.10) mg/L] and FDP variation [(19.29 ± 18.67) mg/L,r was 0.556 and 0.460,respectively;P was 0.020 and 0.002,respectively].Conclusions The D-dimer and FDP levels are kept falling in PE patients after treatment,suggesting that the pulmonary artery embolus is being dissolved.