Objective To explore the role of liver X receptor (LXR) agonist T0901317 on thrombomodulin (TM) expression in human glomerular endothelial cells and the possible mechanisms.Methods Different concentrations of T0901317 were used to stimulate human glomerular endothelial cells for different time,then LXRα,LXRβ expression were detected by using Western blotting analysis;the roles of T0901317 on TM mRNA and TM protein expression were observed by using real-time PCR,Western blotting and immunofluorescence assay.LXRα,LXRβ gene interference segment Si-hLXRα,Si -hLXRβ were transfected into human glomerular endothelial cells with the concentration of 100 nmol/L respectively,then the roles of Si-hLXRα,Si-hLXRβ on the TM protein and TM mRNA expression were assayed by Western blotting and real time PCR.Results Human glomerular endothelial cells expressed LXRα and LXRβ.Compared to the normal cells and DMSO group,T0901317 could significantly promote TM expression in human glomerular endothelial cells (P ＜ 0.05) and showed a time -and dose-dependent manner.TM expression in Si-hLXRα transfected group was significantly lower than that in the control group (P ＜ 0.05),while TM expression in Si-hLXRβ transfected group had no significant difference compared to the control group.Conclusions Human glomerular endothelial cells express LXRα and LXRβ.LXR agonist T0901317 promotes TM expression in human glomerular endothelial cells,which may be mainly through activating LXRa.

ObjectiveTo investigate the effects and molecular mechanism of endoplasmic reticalam stress (ERS) on albumin-induced apoptosis in renal proximal tubular cells (HKCs). MethodsWestern blot was performed to detect the relationship of the expression of glucose-regulated protein 78 (GRF78) and CCAAT/enhancer-binding protein-homologous protein (CHOP) with the action time and concentration of haman serum albumin (HSA). Expression levels of CHPO mRNA and protein in HKCs after CHOP siRNA transfection were examined by real-time fluorescence quantitative PCR and Western blot respectively. Annexin-V-FITC and PI doable staining cytometry was used to detect the apoptosis of HKCs induced by HSA and influenced by CHOP siRNA. Results(1)After HKCs were stimulatde by 0, 5, 10, 20 g/L albumin for 24 hours respectively, the expression of GRP78, CHOP and HKCs apoptosis were increased with the albumin concentration (P<0.01). After HKCs were stimulated by 20 g/L albumin for 0, 6, 12, 24, 36 hours respectively, the expression of GRP78 was up-regulated at 6-hour, while CHOP and HKCs apoptosis were increased at 12-hour, and significant differences were found among groups (P<0.01). (2) CHOP siRNA significantly inhibited albumin-induced HKC CHOP mRNA and protein expression, as well as HKC apoptosis (P<0.01). ConclusionsRenal tubular cells exposed to high protein load result in EBS. ERS may subsequently lead to tubular damage by activation of pro-apoptosis factor CHOP.

Objective To explore the influence of albumin-activated renal tubular epithelial cells (RTECs)on peritubular capillaries in co-culture system and its potential mechanism. Methods Endocytosis of TRITC labeled bovine scrum albumin (TRITC-BSA) by HKC was detected by laser scanning confocal fluorescence microscope. HKC or HKC transfected with cubilin (endocytic receptor of albumin) siRNA or pre-treated with rotenone was incubated with albumin(20 g/L) for 24 h respectively. Fluorescence probe technique and spectrometry were applied for determination of intracellular superoxide anion O2-and H2O2 in supematant. Then, the albumin-aetivated-HKC, pretreated-HKC with cubilin siRNA or rotenone, was cultured with HUVEC for 24 h in co-culture system respectively. HUVEC proliferation was determined by MTT and cellular apoptosis was analyzed by flow cytometry. Tabular morphogenesis of endothelial cells was examinedby microscopy. Results TRITC-BSA uptake was obviously lower in HKC transfected with cubilin siRNA. Intracellular generation of O2-and H2O2 in culture supernatant was increased in dose-and time-dependent manner after stimulating with albumin. The levels of O2-and H2O2 were suppressed by cubilin siRNA and rotenone. In co-culture system, albumin-activated-HKC induced endothelial cells apoptosis and inhibited their capillary tubular morphogenesis. Pretreatment of HKC with cubilin siRNA or rotenone could suppress endothelial cells apoptosis and promote capillary tubular morphogenesis. Conclusions There may be a crosstalk between RTECs and peritubular microvascular endothelial cells in renal proteinurie diseases. The generation of ROS by albumin-activated RTECs may play an important role in this process.

OBJECTIVE: To investigate the curative effects and side effects of hirudin in treating immunoglobulin A nephropathy (IgAN) with hematuria and minimal proteinuria in a short-term. METHODS: Two hundred and sixty-two histologically confirmed cases of IgAN with hematuria and minimal proteinuria from 1998 to 2007 were randomly divided into hirudin-treated group (peroral administration of Maixuekang capsules) and dipyridamole-treated group (peroral administration of dipyridamole). In the two groups, contrast analysis of conformation and counts of erythrocytes in urine, urine protein quantitation in 24 hours, levels of serum creatinine (Scr) and creatinine clearance rate (Ccr), blood lipid, five items of blood clotting and side effects was performed. RESULTS: After six-month treatment, the anisotrophy rate and the counts of erythrocytes in urine, and the urine protein quantitation in 24 hours in hirudin-treated group were decreased distinctly as compared with pre-treatment (P<0.01) and dipyridamole-treated group (P<0.05). On the other hand, Ccr was increased obviously in hirudin-treated group as compared with pre-treatment and dipyridamole-treated group (P<0.01). The blood lipid was also ameliorated in hirudin-treated group, but there was no significant difference. The anticoagulation effect of hirudin was better than dipyridamole (P<0.01). Efficacy assessment showed that the total response rate, complete remission rate and predominance remission rate in hirudin-treated group were higher than those in dipyridamole-treated group. Few side effects were found in both groups, and the rate of adverse reaction in gastrointestinal tract was lower in hirudin-treated group as compared with that in dipyridamole-treated group (P<0.05). CONCLUSION: Compared with dipyridamole, hirudin has superiority in kidney protection and decreasing the anisotrophy rate, counts of erythrocytes in urine and the urine protein.

Objective To observe the effect of recombinant adenovirus-mediated the fms-like tyrosine kinast-1 (FLT-1) gene promoter carrying protein C (PC) gene (Ad-FLT-1/PC) on apoptosis and oxidative stress in rat with diabetic nephropathy atherosclerosis. Methods High sugar, high fat diet and streptozotocin by intraperitoneal injection were used to establish diabetic nephropathy atherosclerosis rat model. The rats were randomly divided into Ad-FLT-1/PC group (n=23), Ad-GFP group (n=23) and normal saline group (n=22) and injected with 300 ul Ad-FLT-1/PC, Ad-GFP and normal saline by caudal vein. Another healthy rats (n=23) were control group. At day 1, 3, 7 and 14 after transfection, rats of each group were randomly executed to observe vascular apoptosis and assay the level of SOD and MDA in plasma by the kit method of WST and TBA. Results Vascular cell apoptosis was observed in Ad-FLT-1/PC group, Ad-GFP group and NS group, the apoptosis index showed no statistical difference between three groups at each time point (P>0.05). At day 14 after transfection, Ad-FLT-1/PC group rats had higher concentration of the plasma SOD and lower MDA than Ad - GFP group and NS group. The difference were statistically significant (all P<0.05). Conclusions The recombinant adenovirus Ad-FLT-1/PC can effectively regulate oxidative stress but not apoptosis.

Objective:To analyze HIV testing behaviors and associated factors in men who have sex with men (MSM) and provide evidence for making intervention on promoting HIV testing.Methods:Our observational study was conducted between April and June, 2018 in Wuxi, Jiangsu province in MSM. A self-completed questionnaire was used to collect the information about their socio-demographic characteristics, sexual behaviors, acceptance of intervention services, psychological status and HIV testing behaviors. Structural equation model (SEM) was chosen to fit data and the model was modified to analyze the relationships between variables.Results:A total of 410 MSM were enrolled in our study, among whom 72.9%(299/410) were aware of the necessity of HIV testing and 69.0%(283/410) reported having HIV tests in the last year. A modified structural equation model presented good fitting results with fitting index of root mean square error of approximation (RMSEA)was 0.065, normed fit index (NFI) was 0.946, relative fit index (RFI) was 0.922, Tucker-Lewis index (TLI) was 0.968 and comparative fit index (CFI) was 0.952. Results of SEM showed that intervention service was the factor with the largest standardized total effect value of 0.57 associated with HIV testing behaviors among MSM. Among intervention services, HIV testing reminding had the biggest factor loading of 0.88. Psychological status scores played a negative role in HIV testing behaviors among MSM with the path coefficient value of -0.33.Conclusions:The awareness and testing rate of HIV among MSM population need to be further improved. While effective measures are implemented to promote HIV testing among MSM, more attention should be paid to the mental health problems of MSM.

Objective:This study aimed to discover the HIV infection and associated factors among male sex workers (MSW) who provide sexual services for men. Data from this study thus can be used to provide evidence for designing related intervention programs.Methods:In this observational study, MSW were recruited from May to July, 2019 in KTV venues in Wuxi, China. A questionnaire was used to collect information with blood sample collected and tested for HIV and syphilis. EpiData 3.0 and SPSS 17.0 software were used to clean up data and statistical analysis.Results:A number of 500 MSW were involved including 243 (48.6 %) heterosexual and 257 (51.4 %) homosexual MSW. The overall HIV prevalence was 5.4 %(27/500) among all the MSW. there were significant differences between the HIV prevalence rates, the heterosexual MSW (3.3 %, 8/243) and the homosexual MSW (7.4 %, 19/257)( χ2=4.112, P=0.043). In the past 3 months, 28.0 % (72/257) of the homosexual MSW mainly engaged in receptive anal sex which was higher than 11.5 %(28/243) of the heterosexual MSW. Compared to 15.6 % (40/257) of the homosexual MSW who engaged in heterosexual behavior, a higher proportion of 98.4 % (239/243) was noticed among the heterosexual MSW. Higher percentage (44.9 %, 109/243) appeared among the heterosexual MSW who had not been tested for HIV than the homosexual MSW (20.6 %, 53/257). Results from logistic regression multivariate analysis showed that age ≥30 (a OR=7.54, 95 %CI: 2.53-37.11), having unprotected anal sexual practice (a OR=3.76, 95 %CI:1.15-12.23), having anal sex after drinking alcohol (a OR=10.91, 95 %CI: 2.29-51.87) and syphilis tested positive (a OR=8.23, 95 %CI:1.29-52.51) were risk factors associated with HIV infection among the heterosexual MSW. Having unprotected anal sexual behavior (a OR=2.94, 95 %CI: 1.17-7.37), having group anal sex (a OR=4.08, 95 %CI:1.05-15.81), without record on HIV testing (a OR=6.58, 95 %CI: 2.01-18.06) and syphilis tested positive (a OR=4.55, 95 %CI: 1.15-18.06) were risk factors associated with HIV among the homosexual MSW. Conclusions:High HIV prevalence was discovered among both heterosexual and homosexual MSW in Wuxi, China. Differences in sexual behaviors between these two groups should be considered when designing targeted HIV intervention programs for these populations.

Objective To explore the application value of cone-shaped gastric tube combined with cervical end-to-end anastomosis in thoracoscopic and laparoscopic esophagectomy for esophageal cancer.Methods The retrospective and descriptive study was conducted.The clinical data of 122 patients with esophageal cancer who were admitted to West China Hospital of Sichuan University from December 2016 to December 2017 were collected.There were 89 males and 33 females,aged (61±8)years,with a range from 48 to 81 years.McKeowntype three-incision esophagectomy was performed,and the cone-shaped gastric tube was pulled up to esophagus in left neck for hand-sewn end-to-end anastomosis after the dissection of esophagus and stomach under total thoracoscopy and laparoscopy.Observation indicators:(1) surgical treatment situations;(2) postoperative complications;(3) follow-up.Follow-up using outpatient examination was performed to detect postoperative gastroesophageal reflux,anastomotic stenosis and evaluate anastomotic width at 1,3,6 months and one year postoperatively up to December 2018.Measurement data with normal distribution were represented by Mean±SD.Measurement data with skewed distribution were described by M (P25,P75) or M (range).Count data were expressed by absolute number.Results (1) Surgical treatment situations:122 patients underwent laparocopic McKeown-type three-incision esophagectomy successfully,using cone-shaped gastric tube combined with cervical hand-sewn end-to-end anastomosis as digestive tract reconstruction,with no intraoperative conversion to open surgery.The operation time,cervical anastomosis time,and volume of intraoperative blood loss were (229 ± 49) minutes,(27± 1) minutes,and 50 mL (40 mL,60 mL),respectively.There were 6-8 stations of lymph node dissected,and the number of lymph node dissected were 19 (15,25).Duration of postoperative hospital stay was 10 days (9 days,11 days) in the 122 patients.(2) Postoperative complications:31 of 122 patients had postoperative complications.The primary complications:3 patients with anastomotic fistula were cured by conservative treatment including enteral nutrition through placement of nutritional tube under gastroscope,closed thoracic drainage and anti-infection;6 cases with severe thoracic gastric dilation were cured after gastrointestinal decompression.The secondary complications of 22 patients included 8 cases with hoarseness caused by recurrent laryngeal never injury,5 with arrhythmia,9 with pulmonary infection.They were cured after symptomatic and supportive treatment.No chylothorax occured,and there was no perioperative death.(3) Follow-up:all the 122 patients were followed up for 10-24 months,with a median time of 19 months.During the follow-up,7 cases with anastomotic stenosis including 4 scoring less than grade 2 and 3 scoring more than grade 3 were relieved after dilation through gastroscope.There were 33 of 122 patients without any reflux symptoms,and 89 with reflux symptoms,among which 52 were scored 1,25 were scored 2 and 12 were scored 3.The width of gastroesophageal anastomosis measured by barium radiography at 1 month after operation was (1.2±0.4) cm.Conclusion Coneshaped gastric tube combined with cervical end-to-end anastomosis in digestive tract reconstruction of thoracoscopic and laparoscopic esophagectomy can reduce the incidence of postoperative anastomotic complications and thoracic gastric dilation,and nasogastric tube placement could be abandoned,which demonstrates good safety and universality.

OBJECTIVETo explore the role of TNFα induced protein 3 interacting protein 1 (TNIP1) in the pathogenesis of myasthenia gravis (MG) among patients with thymoma.

METHODSFrom December 2014 to March 2015, 11 patients with MG associated thymoma (MGT) and 11 non-MG thymoma (NMGT) patients receiving thymectomy were selected. Thymus specimens were obtained during surgery, and peripheral venous blood samples were obtained before the surgery. For the MGT cohorts, peripheral venous blood samples were also collected at 3-6 months after the surgery. TNIP1 mRNA was determined with quantitative real-time PCR (qPCR), and its protein expression was evaluated by Western blotting.

RESULTSFor both thymus specimen and peripheral blood samples, the levels of TNIP1 mRNA and protein in the MGT group were both significantly lower than those of the NMGT patients (P<0.05). For the MGT group, the levels of TNIP1 mRNA and protein of the peripheral blood samples have increased following thymus resection compared with before the surgery.

CONCLUSIONReduced TNIP1 expression may have a role in the pathogenesis of MG for patients with thymoma. Thymectomy may help to recover the expression of TNIP1 among such patients.

Blotting, Western ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Gene Expression ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Myasthenia Gravis ; complications ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Thymectomy ; Thymoma ; complications ; surgery ; Thymus Neoplasms ; complications ; surgery

Objective:To evaluate the extent and progression of coronary artery calcification in maintenance hemodialysis (MHD) patients, and to explore the risk factors of rapid progression of coronary artery calcification in MHD patients.Methods:The patients who underwent MHD in the Huashan Hospital affiliated to Fudan University from January 1, 2013 to December 31, 2017 were enrolled. This study included cross-sectional study and prospective cohort study. Multi-slice spiral computed tomography was used to measure coronary artery calcification, and coronary artery calcium score (CACS) was calculated. In the cross-sectional study, 62 MHD patients were enrolled. According to baseline CACS, the patients were divided into low calcification group (CACS < 100) and high calcification group (CACS ≥ 100). The nutritional and bone mineral metabolism indexes were compared between the two groups. Multiple linear regression analysis was used to analyze the correlation between CACS and muscle mass and laboratory indicators. Since 6 patients were lost to follow-up, 56 MHD patients who were followed-up regularly were enrolled in the prospective cohort study. According to the progression of CACS, the patients were divided into slow progression group (ΔCACS/year < 100) and rapid progression group (ΔCACS/year ≥ 100). Logistic regression equation was used to analyze the risk factors of coronary calcification progression. Hosmer-Lemeshow goodness of fit test and receiver operating characteristic curve were used to evaluate the performance of multivariate logistic regression model.Results:In the cross-sectional study, the age of 62 patients was (62.34±10.82) years old, and the median dialysis age was 78 (39,139) months. Among the 33 male patients, compared with the low calcification group ( n=7), the high calcification group ( n=26) had older age ( t=-2.281, P=0.030) and higher blood triglyceride ( Z=-1.985, P=0.047), and there was no statistically significant difference in muscle mass between the two groups; among the 29 female patients, the muscle mass/height 2 ( t=-2.600, P=0.015) and serum calcium ( t=-2.641, P=0.014) in the high calcification group ( n=15) were both higher than those in the low calcification group ( n=14), and the hemoglobin level was lower ( t=2.531, P=0.018), and the difference in muscle mass between the two groups was not statistically significant. High sensitivity C-reactive protein ( β=0.425, P=0.022) was independently correlated with CACS in male patients, and muscle mass/extracellular water ( β=-0.580, P=0.001) was independently correlated with CACS in female patients. In the prospective cohort study, the age of 56 patients was (59.82±11.14) years old, and the median dialysis age was 82 (40, 146) months. There was no significant difference in all-cause mortality between slow progression group ( n=22) and rapid progression group ( n=34), but the proportion of cardiovascular events in rapid progression group was significantly higher than that in slow progression group ( P=0.017). Compared with the slow progression group, the rapid progression group had higher proportion of males ( χ2=4.791, P=0.029), older age ( Z=-2.131, P=0.038), lower baseline muscle mass/extracellular water ( Z=2.482, P=0.016) and high-density lipoprotein cholesterol ( t=2.133, P=0.042), and faster rate of muscle mass loss (Δmuscle mass·height -2·year -1) ( Z=-2.282, P=0.023). Multivariate logistic regression analysis results showed that muscle mass loss ( OR=0.089, 95% CI 0.010-0.792, P=0.030) and baseline CACS ( OR=1.003, 95% CI 1.000-1.005, P=0.021) were influencing factors for progression of coronary artery calcification in MHD patients. Conclusion:Increasing baseline CACS and rapid reduction in muscle mass are risk factors for the progression of coronary artery calcification in MHD patients.