Objective To investigate the renoprotective effect of irbesartan on tubulointerstitial fibrosis in rats with adriamycin-induced nephropathy and its possible mechanism. Methods Rats received twice-intravenous injections of adriamycin(ADR) after the right kidney was removed. Those rats were randomly assigned to irbesartan treatment group and nephropathy group. Treatment group received 50 mg? kg-1 ? d-1 irbesartan for 4 weeks. Rats with sham operation served as normal control. Proteinuria and serum creatinine of were measured after 4 weeks. Renal histopathological changes were evaluated as well. Immunohistochemistry was used to examine the protein expression of TGF-?1, MMP-9 and TIMP-1. The mRNA levels of MMP-9 and TIMP-1 were detected by in situ hybridization. Results Proteinuria of treatment group decreased significantly as compared to nephropathy group. TGF-?1, TIMP-1 and MMP-9 were significantly lower than that of nephropathy group in tubulointerstitium and consistently associated with tubular degeneration and interstitial fibrosis progressed. Conclusion Irbesartan has a renoprotective effect on tubulointerstitial fibrosis by modulating the ECM degradation.

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 investigate the expression of matrix metalloproteinase-2 (MMP-2)in pancreatic carcinoma and the relationship between MMP-2 with tumour clinicopatholngical features. Methods The expression of MMP-2 was detected by S-P immunohistochemistry in 36 cases with pancreatic carcinoma and 14 normal pancreat-ic tissues. Results The positive expression rate of MMP-2 was 66.7% (24/36) in pancreatic carcinoma tissue and 14.3% (2/14) in normal pancreatic tissues (χ2 = 3. 587, P < 0.01 ) ;The expression rate of MMP-2 in pancreatic carcinoma tissue with positive-node was 86.7% ( 13/15 ) ,which was higher than that with negative-node,which was 52.4% ( 11/21 ) ( P < 0.05 ) ; As to TNM staging in pancreatic carcinoma, the expression rate of MMP-2 was 41.2% (7/17) with Ⅰ,Ⅱ staging and 89.5% (17/19) with Ⅲ,Ⅳ staging(χ2=9.418,P <0. 01 ) ;The expression rate of MMP-2 was 50.0% (5/10) ,66.7% (10/15) and 72.8% (8/11) in highly,moderately and poorly differentiated pancreatic carcinoma(P > 0.05 ). Conclusions The expression of MMP-2 is strengthened significantly in pancreatic carcinoma tissue and involved in turnout invasion and metastasis features; MMP-2 might be regarded as one more marker for the invasive properties of pancreatic carcinoma.

Objective To study the expressions of TGF-?1 and p21WAF1 in middle ear cholesteatoma and discuses their effects on cholesteatoma.Methods The expressions of TGF-?1 and p21WAF1 were determined by immunohistochemical S-P technology and computer image analysis in 20 specimens of cholesteatoma epithelium and 10 specimens of normal external canal skin.Results ① p21WAF1 and TGF-?1 could be detected in middle ear cholesteatoma and in normal ear canal skin.p21WAF1 was located at karyon,taking on yellow-brown pellet;TGF-?1 was located at cytoplasmic,taking on yellow-brown pellet.②The p21WAF1 positive rate in middle ear cholesteatoma was 65%;and there were almost no positive cells in normal external ear canal skin.p21WAF1 positive cells were detected in each layers,its LI was 28.9%?6.7%.Compared with normal ear canal skin,the expression level of p21WAF1 in middle ear cholesteatoma epithelium was higher than that in normal ear canal skin(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 investigate the inhibitory effects of RNA interference on expression of matrix metalloproteinase-2(MMP-2)gene and invasiveness of human pancreatic cancer cell line PANC-1 in vitro.Methods Small interference RNA targeting MMP-2 gene was designed and constructed to plasmid pGCsi-U6.Recombinant plasmids were transfected to pancreatic carcinoma PANC1 cells with Lipofectamine 2000.The efficiency of transfection was evaluated by flow cytometry.RQPCR was used to detect the expression of MMP-2 mRNA.The expression of MMP-2 protein was determined by ELISA.The invasiveness of PANC-1 cells was measured by transwell chamber experiment.MTT assay was used to detect the proliferation and growth of PANC-1 cells.Results Sequencing confirmed that the MMP-2 siRNA plasmid was successfully constructed.The best efficiency of transfecting recombinant plasmid was 82.1%.After transfection of the MMP-2 siRNA plasmid, the MMP-2 gene expression of PANC-1 cells was suppressed to 71.74 %(P＜0.05),and protein expression of MMP-2 fell to 49.82%(P＜0.05).The corresponding inhibition ratio of invasiveness was 33.0%(P＜0.05).There was no marked difference in proliferation rate measured by MTT assay among different groups(P＞0.05).Conclusions RNAi targeting MMP-2 can suppress invasiveness of PANC-1 cells in vitro.This suggests that MMP-2 could be a target for gene therapy of pancreatic carcinoma.RNAi is expected to open up a new prospect for tumor therapy.

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 determine the distribution of allergens in patients with allergic rhinitis in Wenzhou area. METHODS Patients with AR symptoms from January 2013 to December 2014 were given skin prick test (SPT). The clinical data about SPT was retrospectively colleceted to analyze SPT results. RESULTS 1. Among 2991 individuals, the total positive rate of SPT was 82.0%, with Dermatophagoides farinae (Der. f) and Dermatophagoides pteronyssinus (Der.p) as the most common allergens; the positive rate of inhalant allergen was obviously higher than that of ingestive ones, with significantly statistical difference(χ2=2006.557,P<0.01). Most of patients were allergic to double allergens; the intensity of Der.f and Der.p mainly presented as (++++), with no significant difference(Z=-0.391, P=0.696). 2. There was significant difference of variation with seasons(χ2=34.254, P<0.01). 3. No significant difference of positive rate were observed in different AR-courses(χ2=16.102, P<0.01). 4. Significant difference of positive rate was found among different age-groups; The positive rate was increased along with growing age, got peak at group of 10-12 years old, and then got down after that. CONCLUSION Dust mite was the main allergen coursed AR in Wenzhou area. Seasons and age were two important factors effecting on positive of SPT and onset of AR. The positive rate of allergens was related to age.

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.