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 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.

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 investigate the clinical efficacy of gastroscopic 'sandwich’ injection and sequential laparoscopic splenectomy combined with pericardial devascularization in the treatment of gastric variceal bleeding.Methods:The retrospective cohort study was conducted. The clinical data of 52 patients with cirrhotic portal hypertension combined with gastric variceal bleeding who were admitted to Affiliated Hospital of Jiaxing University between March 2011 and October 2019 were collected. There were 33 males and 19 females, aged (63±9)years, with a range of 41-83 years. Of the 52 patients, 31 undergoing gastroscopic 'sandwich’ injection and sequential laparoscopic splenectomy combined with pericardial devascularization and 21 undergoing laparoscopic splenectomy combined with pericardial devascularization were allocated into sequential group and laparoscopic group, respectively. Observation indicators: (1) surgical situations; (2) complications; (3) changes in gastric varices after treatment; (4) follow-up. Follow-up was performed using telephone interview combined with outpatient examination to detect survival of patients up to December 2019. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed by the t test. Measurement data with skewed distribution were represented as M (range), and comparison between groups was analyzed by the Mann-Whitney U test. Count data were expressed as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Ordinal data was analyzed by nonparametric rank sum test. Results:(1) Surgical situations: patients of sequential group and laparoscopic group underwent surgery successfully. The operation time and volume of intraoperative blood loss of the sequential group were (112±16)minutes and (57±11)mL, respectively, versus (103±14)minutes and (55±9)mL of the laparoscopic group, showing no significant difference in the above indicators between the two groups ( t=0.963, 1.052, P>0.05). (2) Complications: 11 patients in the sequential group had postoperative complications including 1 case with perioperative bleeding, 2 cases with postoperative gastrointestinal rebleeding, 4 cases with ascites, 4 cases with portal vein thrombosis. There was no death in the sequential group. Sixteen patients in the laparoscopic group had postoperative complications including 2 cases with perioperative bleeding, 6 cases with postoperative gastrointestinal rebleeding, 4 cases with ascites, 4 cases with portal vein thrombosis. Three patients died in the laparoscopic group. There was no significant difference in the cases with perioperative bleeding or cases with ascites between the two groups ( P>0.05) and no significant difference in the cases with portal vein thrombosis or death between the two groups ( χ2=0.082, 0.082, P>0.05). There was a significant difference in the cases with postoperative gastrointestinal rebleeding between the two groups ( P<0.05). Cases with postoperative gastrointestinal rebleeding, cases with ascites, cases with portal vein thrombosis in the sequential group were cured after the treatment of gastroscopy, low salt diet combined with diuretic or low dose warfarin, respectively. Of the 6 patients with postoperative gastrointestinal rebleeding in the laparoscopic group, 3 were cured after the treatment of gastroscopy and 3 died due to failure to rescue in time. Cases with ascites and cases with portal vein thrombosis in the laparoscopic group were cured after the treatment of low salt diet plus diuretic or low dose warfarin, respectively. (3) Changes in gastric varices after treatment: at postoperative 6 months, 31 patients in the sequential group were diagnosed with negative gastric varices; 15 of 21 patients in the laparoscopic group were diagnosed with negative gastric varices, 3 cases were diagnosed with obvious gastric varices and 3 cases were diagnosed with severe gastric varices. There was a significant difference in the cases with gastric varices between the two groups ( Z=-3.128, P<0.05). At postoperative 12 months, 29 patients in the sequential group and 13 patients in the laparoscopic group were diagnosed with negative gastric variceal. There were 2 patients in the sequential group diagnosed with obvious gastric varices, and 8 patients in the laparoscopic group diagnosed with gastric varices including 3 cases with obvious gastric varices and 5 cases with severe gastric varices. There was a significant difference in the cases with gastric varices between the two groups ( Z=-2.933, P<0.05). Cases with obvious gastric varices in the sequential group were cured after the treatment of gastroscopy. Cases with obvious or severe gastric varices in the laparoscopic group were cured after the treatment of gastroscopy except 1 died due to massive gastrointestinal hemorrhage. (4) Follow-up: 52 patients were followed up for 1-8 years, with a median time of 4 years. All the 31 patients in the sequential group and 18 ptients in the laparoscopic group survived. Conclusion:Gastroscopic 'sandwich’ injection and sequential laparoscopic splenectomy combined with pericardial devascularization in the treatment of gastric variceal bleeding has low recurrence rate of varicosity and low incidence of rebleeding.

Effects of heat treatment conditions (including temperature and time) on the shape memory recovery and corrosion resistance of NiTi self-expanding vascular stents were studied based on working mechanism and clinical use. The
Alloys ; Corrosion ; Hot Temperature ; Materials Testing ; Stents ; Surface Properties ; Temperature ; Titanium

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.