Objective To evaluate the clinical effect and safety of transvaginal lesion resection for cesarean scar pregnancy (CSP).Methods A total of 30 patients with CSP in our hospital received the transvaginal CSP lesion resection from January 2013 to January 2016 and were followed-up to observe the clinical efficacy and safety.Results The operations were successfully completed except one case of conversion to open surgery.The operation time was 30-120 min (53.4±26.0 min), the intraoperative hemorrhage was less than 50 ml, the hospital time was 3-7 d (4.3±1.3 d), and the postoperative time of serum β-hCG decreased to normal value was 14-36 d (24.1±13.2 d).No continuous pregnancy or complications such as incision non-healing, infection, or bladder injury after operation.Conclusion The clinical effect of vaginal CSP lesion resection is good and has fewer complications, being worthy of application.

Aim To investigate the proliferative effect and the apoptosis of rat epididymal epithelial cells induced by podophyllotoxin and its underlying mechanisms.Methods Primary epididymal epithelial cells were cultured in vitro.CCK-8 assay was used to analyze proliferation of epididymal epithelial cells induced by podophyllotoxin on 24, 48 and 72 h.The ultra structural changes of the epididymal epithelial cells were observed by transmission electron microscope.AnnexinV-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell population.The TdTmediated dUTP nick end labeling(TUNEL) technique was applied to observe the morphological changes of apoptotic cells.The expression of tumor necrosis factor alpha(TNF-α) mRNA was investigated by real-time RT-PCR.The level of TNF-α in cell culture supernatant was measured by enzyme-linked immunosorbent assay(ELISA) technology.Western blot was per-formed to determine the protein expression of cytochrome C, caspase-3, caspase-8, caspase-9.Results Podophyllotoxin significantly inhibited the activity of proliferation and induced apoptosis of epididymal epithelial cells in a dose-and time-dependent manner(P<0.05), with a 50％ inhibitory concentration(IC50) value and corresponding 95％ confidence intervals(CI) of 59.36(15.50～227.41), 0.37(0.080～1.70), 0.077(0.017～0.35) μmol·L-1 at 24, 48 and 72 h, respectively.Podophyllotoxin induced cell volume turned round and cell nuclear fragmentation and mitochondrial vacuolation.RT-PCR and ELISA results showed that podophyllotoxin improved the expression of TNF-α mRNA and protein.Western blot results demonstrated that podophyllotoxin activated the protein expression of cytochrome C, caspase-3, caspase-8 and caspase-9.Conclusion Podophyllotoxin can induce rat epididymal epithelial cell apoptosis through both the mitochondria-regulated intrinsic pathway and the TNF receptor-mediated extrinsic pathway.

Objective To investigate the cellular immunologic response of TH 17/Treg cells in the peripheral blood of pelvic tuberculosis patients and explore their roles in the pathogenesis of pelvic tuberculosis.Methods The intracellular flow cytometry was performed to evaluate the expressions of TH 17 and Treg cells in 46 pelvic tuberculosis patients and 25 healthy controls in childbearing age.Twenty-eight of the 46 pelvic tuberculosis patients were followed up to monitor the variation of the TH17/Treg cells after 3 months and 6 months of anti-tuberculosis treatment.Results The percentage of TH 17 cells in the peripheral blood of pelvic tuberculosis patients was (3.26 ± 1.30) ％ which was significantly lower than that of healthy controls [(4.92 ± 1.71) ％,P ＜ 0.01].The percentage of Treg cells in the patients was (5.18 ± 1.53) ％ which was significantly higher than that of healthy controls [(3.26 ± 1.10) ％,P ＜ 0.01].The percentage of TH17 cells in the pelvic tuberculosis patients after 6 months of treatment was (4.67 ± 1.75) ％ which was significantly higher than that in the patients before treatment and after 3 months treatment [(3.26 ± 1.30) ％,P ＜ 0.01 and (3.70 ± 1.06) ％,P ＜0.01,respectively].The percentage of Treg cells in pelvic tuberculosis patient after 6 months of treatment was (3.93 ±0.94)％ which was significantly lower than that in the patients before treatment and after 3 months of treatment [(5.18 ± 1.53)％,P ＜0.01 and (4.94 ± 1.51) ％,P ＜ 0.01,respectively].The percentage of Treg cells in the patients after 6 months of treatment was still significantly higher than that of controls (P ＜ 0.05).The TH 17/Treg ratio before treatment was significantly lower than that of healthy controls (P ＜ 0.01),and the TH 17/Treg ratio was increased after 3 months of treatment but it did not show significant difference compared with that before treatment.The TH 17/Treg ratio after 6 months of treatment (1.18 ± 0.34) ％ was significantly increased in contrast to those after 3 months of treatment and before treatment [(0.77 ± 0.21) ％,P ＜ 0.01 and (0.55 ± 0.13) ％,P ＜ 0.01,respectively].The TH 17/Treg ratio could not rise to the normal level even after 6 months of treatment.Conclusion Both the TH 17 and Treg cells may involve in the immunologic responses of pelvic tuberculosis patients and the imbalance of TH1T/Treg cells may remain persistently.

Objective To evaluate the diagnostic value of two tuberculosis-specific IFN-γ release assays in latent tuberculosis infection among HIV-infected individuals. Methods The levels of tuberculosis antigen-specific IFN-γin 102 HIV patients from AIDS Outpatient Clinic of Shenzhen Third People's Hospital were detected by in-house tuberculosis-specific IFN-γ ELISpot assay and commercial T-SPOT TB kit, and tuberculin skin test (TST) were done at the same time. There were 66 males and 36 females,and the average age was 35. Results Seventeen HIV infected patients were positive in both IFN-γ ELISpot and T-SPOT TB methods, the sensitivity, specificity positive predictive value(PPV), negative predictive value(NPV) and compliance rates of ELISpot were 94. 4% ,94. 0% ,77. 3% ,98. 8% and 94. 1% ,respectively. Three patients were positive in both IFN-γELISpot and T-SPOT TB methods, the sensitivity, specificity, PPV, NPV and compliance rates of TST were 16. 7%, 98. 8%, 75.0%, 84. 7% and 84. 3%, respectively. The average number of spots using three kinds of antigen ESAT-6, Pool A,Pool B obtained were 26. 89 ±5. 77,18. 96 ±4. 75 and 14. 51 ± 3.77, respectively. Only ESAT-6 and Pool B have a statistically significant difference (H=7.557,P = 0.022 9), no significant difference was shown between other groups. There was no significant difference between the positive rate and the CD4+ T cellls number(x2 =0. 860 8 ,P =0. 650 2) ,as the same as the T-SPOT TB (x2 = 1. 396 4, P = 0. 497 5 ). Conclusions The performance of this in-house tuberculosis-specific IFN-γ ELISPot assay was comparable to T-SPOT assay in diagnosis of latent tuberculosis infection, and the sensitivity and specificity of both these two assays were all much higher than TST. They canbe recommended in diagnosing latent tuberculosis infection in HIV infected patients.

Objective:To systematically analyse the blood coagulation features of coronavirus disease 2019 (COVID-19) patients.Methods:An electronic search in PubMed, Scopus, Web of Science, EMbase, and CNKI to collect studies related to the blood coagulation features of COVID-19 patients from 1 January 2020 to 1 May 2020. Two reviewers independently screened literatures, extracted data and assessed the risk of bias of included studies. Then, the platelet count, D-dimer value, prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen of patients with different types of diseases were analyzed by using Stata12.0 software.Results:Thirty-nine retrospective studies involving 6 994 COVID-19 patients were included. The results of meta-analysis showed that:(1) compared with severe group, the platelet count (Weighted mean difference; WMD=20.11, 95% CI 11.53 to 28.69, P<0.001) and APTT (WMD=1.30, 95%CI 0.31 to 2.30, P=0.01) were found to be higher while D-dimer (WMD=-0.41, 95%CI-0.58 to-0.24, P<0.001), fibrinogen (WMD=-0.58, 95% CI-0.76 to-0.39, P<0.001) and PT (WMD=-0.51, 95%CI-0.92 to-0.10, P<0.001) were lower in mild group; the platelet count (WMD=-14.75, 95% CI-29.73 to-0.23, P=0.044) was found to be lower while D-dimer (WMD=1.06, 95% CI 0.65 to 1.47, P<0.001) was found to be higher in critical ill patients. (2)Compared with the survival group, the patients in death group displayed elevated levels of D-dimer (WMD=6.86, 95% CI 4.15 to 9.57, P<0.001) and PT (WMD=1.37, 95% CI 0.73 to 2.02, P<0.001) while platelet count (WMD=-36.40, 95% CI-63.23 to-9.58, P=0.008) remained low. Conclusion:Coagulation dysfunction was common in severe, critical ill and dead COVID-19 patients. Platelet count, D-dimer and PT levels were associated with the severity of the disease, and thus could be used as early warning indicators for the deterioration of the disease during hospitalization.

ObjectiveTo evaluate the IL-17 expression in HIV/tuberculosis-coinfected patients and its role in the pathogenesis of this coinfection.MethodsFifty-four HIV infected patients were divided into three groups:simple HIV infected group,HIV with latent tuberculosis infection (HIV+ LTBI) group and HIV coinfected with active tuberculosis (HIV+ ATB) group.The whole blood intracellular cytokine staining was performed and samples were then detected by BD FACSCanto.The expressions of CD4+ IL-17+ T cells and CD4+ IFNγ+ T cells were analyzed using FACSDiva software.Comparison between groups was done by independent sample t test.ResultsThe CD4+ T cell count and viral load among these three groups were comparable.There were no significant difference of the expression of CD4+ IL-17+ T cells between simple HIV infected group and HIV+ LTBI group (1.40 ± 1.01) ％ vs (1.29±0.86) ％,(t=0.336,P＞0.05),but both of these two groups were much higher than HIV+ATB group (t=3.680,t=2.516,P＜0.05).There were no significant differences of the expression of CD4+ IFNγ+ T cells among these three groups [(32.8±24.0)％ vs (40.3±1 21.9) ％ vs (46.1±31.2)％,(t=-0.939,t=-1.602,t=-0.646,P＞0.05)].ConclusionThe Th17 response is down-regulated in HIV/tuberculosis-coinfected patients,which may play an important antitubercular role in the pathogenesis of coinfection.

Objective To assess the validity of a newly developed in-house ELISPOT IFN-γ release assay (IGRA) for the detection of latent tuberculosis infection among HIV infected individuals. Methods In-house ELISPOT assay were performed, together with a tuberculin skin test in 205 health controls and 110 HIV infected individuals , who had no signs of active tuberculosis at time of enrolment . Results Using the ELISPOT assay, positivity rates for the 205 health controls, 110 HIV infected individuals and 47 AIDS patients on highly active antiretrovial therapy (HAART) were 7. 3% , 24.5% , 29. 8% , respectively. These results indicated that the positive rates obtained from HIV infected individuals (include patient on HAART) was significantly higher than health controls( P ＜ 0.001). We found no significant correlation between the CD4 cell count and positivity of ELISPOT assay (P ＞0.05 ). The proportion of subjects with a positive response to ELISPOT assay were higher than the proportion of tuberculin skin test(TST) responders(P＜0.0001) in HIV infected individuals. Conclusion Our study indicates that IGRA using M. tuberculosis specific antigens are likely to retain their validity for the diagnosis of LTBI among HIV positive individuals.

Objective: To explore the effect of hyperbaric oxygen preconditioning with different frequency on the survival rate of flap and ischemia-reperfusion injury in rats after transplantation, and to explore the best preconditioning conditions to improve the survival rate of rat flaps after transplantation. Methods: Thirty-six Sprague Dawley rats were randomly divided into four groups according to the random number table method, 9 groups in each group.Four groups of rats were pretreated with hyperbaric oxygen pretreatment for 0, 2, 4, and 6 days before the operation, control group, pretreatment 2 d group, pretreatment 4 d group, and pretreatment 6 d group. Taking the midline of the back of the rat as the axis, an ultra-long random flap with a pedicle at the tail end and about 1 cm from the superior iliac spine was designed and cut to a size of 10.0 cm×2.5 cm. The survival of the flaps in each group was observed and the final survival area and survival rate of the flaps were measured on the 7th day after surgery. On the 7th day after operation, the tissue was taken at a distance of 5 cm from the pedicle, and the histopathology was observed; The content of superoxide dismutase (SOD) and malondialdehyde (MDA) in flap tissue was detected by immunohistochemistry, and the expression rate of positive cells in each group was calculated. Immunofluorescence was used to detect the expression of interleukin-6 (IL-6) in the flap tissue. Results: On the 7th day after the operation, the survival area and survival rate of the transplanted flaps in the hyperbaric oxygen pretreatment group were significantly higher than those in the control group (P<0.05), and the pretreatment 4 d and 6 d groups were significantly higher than the pretreated 2 d group (P<0.05), but there was no significant difference between the pretreated 4 d group and the 6 d group (P=0.095). Pathological observation on the 7th day after operation showed that there was some necrosis in the control group, the vascular cells in the pretreated 2 d group had more vascular structures, and more neovascularization was observed in the pretreated 4 d group. The inflammatory cells were the least in the 6 d pretreatment group, and the neovascularization was the same as the pretreatment 4 d group. The absorbance A value of SOD in the control group was 0.009 7±0.000 3, and the positive expression rate was 20%, which was significantly lower than that in the hyperbaric oxygen pretreatment group. The difference was statistically significant (P<0.05). However, the absorbance A value of MDA in the control group was 0.055 1±0.003 0, and the positive expression rate was 55%, which was significantly higher than that in the hyperbaric oxygen pretreatment group. The difference was statistically significant (P<0.05). Among them, the SOD absorbance A value of the pretreated 2 d group was 0.023 8±0.003 0, and the positive expression rate was 30%, which was lower than the pretreatment 4 d group (absorbance A value 0.046 9±0.003 0, positive expression rate 35%) and 6 d group (absorbance A value 0.047 2±0.003 6, positive expression rate 40%), The MDA absorbance A value of the pretreated 2 d group was 0.037 2±0.003 2, and the positive expression rate was 30%, which was higher than the pretreatment 4 d group (absorbance A value 0.014 7±0.002 4, positive expression rate 5%) and 6 d group (absorbance A value 0.017 0±0.001 8, positive expression rate 10%), the difference was statistically significant (P<0.05). However, there was no significant difference in the expression of SOD and MDA between the pretreated 4 d group and the pretreated 6 d group (P>0.05). The expression of IL-6 in the hyperbaric oxygen pretreatment group was significantly lower than that in the control group (absorbance A value 44.937 0±0.594 0), the difference was statistically significant (P<0.05). The absorbance A value in the pretreated 2 d group was 41.698 0±0.724 0, which was significantly higher than the pretreatment 4 d group (absorbance A value 34.049 0±0.323 0) and 6 d group (absorbance A value 33.524 0±0.639 0). The difference was statistically significant (P<0.05). There was no significant difference in the expression of the pretreated 4 d group compared with the 6 d group (P=0.068). Conclusions Hyperbaric oxygen preconditioning can significantly promote the survival of rat flaps after transplantation. Preoperative hyperbaric oxygen preconditioning once daily for 4 consecutive days, can enhance the tolerance of flap tissue to ischemia and anoxia and reduce tissue ischemia-reperfusion injury.

Objective To establish an allogeneic rat model of endometriosis and to evaluate the effects of gonadotropin-releasing hormone (GnRH) agonist GenSci006 on experimental rat endometriosis. Methods Endometrium from SPF grade donor female SD rats were transplanted onto the abdominal wall of recipient female rats to construct an allogeneic endometriosis model. The rats undergoing sham surgery were divided into the sham group. Three weeks later, the length, width and height of the ectopic endometrium were measured, and the volume of the endometrium (V₁) was calculated before drug administration. The modeling rats were randomly divided into four groups: model group, triptorelin group (0.25 mg/kg), GenSci006-1 group (0.125 mg/kg) and GenSci006-2 group (0.25 mg/kg). Each group had 16 rats and received a single dose of the corresponding drug. The sham group and model group were administered an equal volume of solvent. Three weeks after administration, ectopic endometrium was measured to calculate the volume V₂ and inhibition rate. The effect of GenSci006 on rat uterus and ovarian tissues was assessed by comparing organ coefficients and changes in pathological sections. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of serum estradiol (E₂), progesterone (P₄), follicle stimulating hormone (FSH), and luteinizing hormone (LH). Real-time fluorescent quantitative PCR was used to detect the expression of GnRH receptor (GnRHR) mRNA in the hypothalamus and pituitary. Western blot was used to detect the expression of estradiol receptor alpha (ERα), beta (ERβ) and progesterone receptor (PR) in ectopic endometrium. Results Three weeks after administration, compared with the model group, the body weight of rats in the triptorelin and GenSci006-2 groups significantly increased (P < 0.05), while the volume of ectopic endometrium significantly decreased (P < 0.05). Compared with the sham group, the model group showed no significant changes in uterine and ovarian organ coefficients or endometrial thickness (P > 0.05). Compared with the model group, the uterine organ coefficients and endometrial thickness were significantly reduced in the triptorelin and GenSci006-2 groups (P < 0.05). Compared with the sham group, the serum levels of E₂, P₄, FSH and LH in the model group showed no significant changes (P > 0.05). Compared with the model group, the ovarian organ coefficient and serum P₄ levels of rats in the Triptorelin, GenSci006-1, and GenSci006-2 groups were significantly reduced (P < 0.05), while the serum LH levels of rats in the GenSci006-1 group were significantly increased (P < 0.05). However, there were no significant changes in serum E₂ and FSH levels in each group (P > 0.05). Compared with the model group, the expression levels of GnRHR mRNA in the pituitary tissue of rats in the triptorelin and GenSci006-2 groups were significantly downregulated (P < 0.05), with no significantly changes in the hypothalamus (P > 0.05). There were no significant changes in the expression level of GnRHR mRNA in the hypothalamus or the protein levels of ERα, ERβ and PR in the ectopic endometrial tissue in any group (P > 0.05). Conclusion The allogeneic endometriosis rat model is a suitable animal model for screening and evaluating drugs for treating endometriosis. The volume of ectopic endometrium, inhibition rate, uterine and ovarian organ coefficients, and serum E₂ levels may serve as indicators for detecting drug efficacy.