Objective To investigate whether the signaling pathway of PI3K-AKT is involved in Ang II-induced apoptosis in rat renal tubular epithelial cells (NRK-52E).Methods Cultured cells were incubated in media containing either buffer (control) or different concentrations of Ang II (10-9 to 10-6 mol/L) for 24 h. Cells were stained by Annexin V-F1TC/PI Kit and apoptosis was evaluated by flow cytometer. Western blotting was performed to assess the levels of PI3K, total- and phosphor-AKT. Results Ang II induced tubular epithelial cell apoplosis in a dose dependent manner. Ang II significantly inhibited the phosphorylation of AKT. The level of AKT phosphorylation was negatively correlated with apoptosis in Ang II-stimulated cells(r=-0.90 ,P

Objective To evaluate the effects of high glucose on autophagy and apoptosis of podocyte and explore the signaling pathway in high glucose-induce podocyte autophagy.Methods Differentiated mouse podocytes were exposed to high glucose(30 mmol/L) or rapamycin (autophagy enhancer,1 μg/L) or LY294002 (a selective PI3K inhibitor,50 mmol/L) for 24 h.The formations of autophagy were observed by electron microscopy and acridine orange staining.Apoptosis was evaluated by flow cytometry.The expression of autophagy protein LC3-Ⅱ/Ⅰ and Beclin-1 as well as the phosphorylation of AKT and mTOR were examined by Western blotting analysis.Results High glucose induced podocytes apoptosis,increased autophagy and the expression of autophagy-associated proteins (all P ＜ 0.05).Rapamycin further increased the expression of LC3-Ⅱ and Beclin-1 protein (all P ＜ 0.05),but LY294002 inhibited partialiy the protein expression of LC3-Ⅱ and Beclin-1 induced by high glucose (both P ＜ 0.05).Treatment with rapamycin increased the phosphorylation of AKT,but reduced that of mTOR in podocytes.Moreover,LY294002 inhibited phosphorylation of both AKT and mTOR (both P ＜ 0.05).Conclusions High glucose promotes podocyte autophagy and apoptosis.High glucose-induced autophagy is mediated partly through PI3K-AKT-mTOR signaling pathway.

Objective To analyze the pathogenesis and prevention and treatment of limb deep venous thrombosis complicated by postoperative gynaecological malignant tumours.Methods A retrospective analysis of 22 cases with limb deep venous thrombosis complicated by postoperative gynaecological malignant tumours in our hospital were analyzed.According to the situation of patients,surgery or conservative treatment was conducted.The clinical efficacy after treatment was observed.Results In 22 cases,6 cases underwent surgical treatment,16 cases underwent conservative treatment.After treatment,18 cases were cured,3 cases were effective,1 case ineffective.After treatment,the TT,Pt,APTT and FBG levels were (24.17 ±2.13)s,(18.25 ±2.13)s,(31.68 ± 10.30)s and (2.26 ±0.52) g/L,which were all better than before treatment,the differences were statistically significant (t =10.362,9.774,12.281,8.462,P =0.028,0.032,0.012,0.029).Followed up for 3 months,no recurrence of LDVT was observed.Conclusion The cases of gynecological malignant tumor have higher risk of limb deep vein thrombosis,we should taken prevention actively,once diagnosed should take effective intervention measures to improve the prognosis.

Objective To evaluate the effects of autophagy on oxidative stress induced by contrast media in podocytes.Methods The differentiated mouse podocytes were exposed to contrast media (Iopromide,50 mg/L)、rapamycin (Rap,autophagy enhancer,1 ng/L),3-methyladenine (3-MA,autophagy inhibitor,2 mmol/L) for 2 hours.The expression of autophagy protein LC3-Ⅱ and Beclin-1 as well as oxidative stress-related proteins Catalase,MnSOD were detected by Western blot.The formations of autophagy were observed by MDC staining,and the levels of reactive oxygen species (ROS) by CM-H2DCFDA staining.Cell activity was evaluated by CCK8 assay.Results Both the levels of oxidative stress and autophagy in podocytes increased when stimulated by contrast media,the expression of LC3-Ⅱ and Beclin-1 were enhanced,Catalase and MnSOD were inhibited (all P ＜ 0.05).Rapamycin increased the expression of Catalase,MnSOD and cell activity of podocytes,reduced the generation of ROS (all P ＜ 0.05),but in Rap group,cell activity showed no significant difference (P ＞ 0.05).3-MA decreased the expression of Catalase 、MnSOD and inhibited the cell activity of podocyte,increased the generation of ROS (all P ＜ 0.05).Conclusion Autophagy protects podocyte from contrast media by the means of reducing oxidative stress.

Objective To investigate the characteristics and outcome of anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) in patients with renal injury. Methods AAV patients with renal injury diagnosed in the Department of Nephrology, Renmin Hospital of Wuhan University, from January 2012 to January 2017 were included into this study. Patients were divided into MPO-ANCA positive and PR3-ANCA positive groups for further study. The clinical characteristics, pathological and laboratory indexes, treatment and prognosis were retrospectively analyzed. Results A total of 68 cases were enrolled, among which 52 cases (76.5%) were MPO-ANCA positive and 16 cases (23.5%) were PR3-ANCA positive, and 41 patients (60.3%) were over 65 years old. The incidences of interstitial lung disease, digestive and nervous system damage in PR3-ANCA positive group were significantly higher than those MPO - ANCA positive group (P<0.05). There were significant differences of hemoglobin, complement C3, complement C1q, IgE, 24 h urinary protein,erythrocyte sedimentation rate, procalcitonin, BVAS score and eGFR in two groups (P<0.05). 19 cases had done renal biopsy ,among them 14 cases were MPO-ANCA positive and 5 cases were PR3-ANCA positive. Incidence of crescentic necrotizing glomerulonephritis in PR3-ANCA positive group was significantly higher than that in MPO - ANCA positive group, and incidence of diffuse global glomerulosclerosis in MPO-ANCA positive group was significantly higher than that in PR3-ANCA positive group (all P<0.05). At the median follow-up time of 32 months, the relapse rate at 6 month of MPO-ANCA-positive and PR3-ANCA-positive patients were 46.2% and 75.0%, respectively (P<0.05). Multivariate logistic regression analysis showed that PR3-ANCA positive, age≥65 years old, baseline eGFR<30 ml·min-1·(1.73 m2)-1, and combined with pulmonary interstitial lesions were all independent risk factors for relapse. And the incidence of ESRD were 42.3%and 75.0%during the follow-up period and 10 patients (14.7%) died. COX regression analysis showed that patients older than 65 years old, BVAS score≥18 points, eGFR<30 ml·min-1·(1.73 m2)-1 and complicated with pulmonary interstitial disorders at the onset were independent risk factors causing ESRD or death. Conclusion The PR3-ANCA-positive patients had more severe renal injury than those with MPO-ANCA-positive patients, and the injury of extrarenal organs was more serious, recurrence rate was higher, and the prognosis was worse.