Objective To investigate the effect of leflunomide and prednisone in treating refractory nephropathy syndrome.Methods The 60 refractory nephropathy patients in our hospital from Oct,2000 to May,2003 were divided into two groups at random.Paients in test group received leflunomide and prednisone.Patients in control group received mycophenolate mofetil and prednisone.Clinical data were observed in the 2nd,4th,6th,8th,12th,16th,20th,24th and 28th week.Results After receiving leflunomide therapy in the test group,the proteinuria was decreased significantly(P0.05),but rate of showing efficacy in 12th week in Leflunomide group was higher than mycophenolate mofetil group(P

Objective To investigate the efficacy and safety of mycophenolate mofetil combined with methylprednisolone and lamivudine on the treatment of hepatitis B virus associated glomerulonephritis(HBV-GN).Methods Twenty-four patients with hepatitis B virus associated glomerulonephritis were confirmed by renal biopsy and immunohistochemistry,these participant patients were admitted to the Third Affiliated Hospital of Sun Yat-Sen University from Jan,1999 to Jan,2004.They were treated by MMF combined with methylprednisolone and lamivudine.The initial dosage of MMF was 1.0~1.5 g/d.Methylprednisolone at the dosage of 0.4 mg/(kg?d)was used at the beginning of the combined treatment.Lamivudine was in the dosage of 0.1 g/d.The duration of the treatment was six months.Regular test was conducted every two weeks.Results Nine cases had fully remission,11 cases had partial remission and 4 cases had no efficiency;no patient deterioration.Renal and hepatic function remained stable,blood cell didn't decrease and the reproduction of HBV didn't increase during the treatment.Conclusion MMF combined with methylprednisolone and lamivudine is an effective and safe method for HBV-GN.

Objective To investigate the serum leptin levels of chronic hemodialysis patients and its relation to their nutritional status.Methods Twenty-nine maintenance hemodialysis patients were included in the study.TSF(triceps skin fold),BMI(body mass index) and FAT%(content of fat),lymphocyte count,serum albumin,globulin,total iron binding capacity,BUN,creatinine,cholesterol,triglyceride and leptin were measured.Malnutrition-inflammation score(MIS) was used to assess the patients nutritinal status.Results Levels of leptin were positively correlated with BMI,FAT%,TSF and MIS(P

[Objective] To investigate the effects and mechanisms of Nrf2-ARE (nuclear factor erythroid-2 related factor-anti-oxidant response element) pathway on uremic serum-mediated endothelial dysfunction in human aortic endothelial ceils.[Methods] Human aortic endothelial cells were incubated in endothelial cell medium containing 10％ normal serum,10％ non-diabetic nuremic serum or 10％ diabetic uremic serum respectively,and 20 μmol/L tertiary butyl hydroquinone (tBHQ) were pretreated with cells to active Nrf2-ARE pathway.The cells apoptosis rate were measured by flow cytometry,and the synthesis of NO was detected by flow cytometry and immune fluorescent confocal,while the expression of P-eNOSer1177/eNOS,and quinone oxidoreductase-1 (NQO1) were measured by western blotting.The levels of malondialdehyde,superoxide dismutase,eatalase,and glutathione in these cells were also measured with kits.[Results] Aortic endothelial cells incubated with uremic serum had a higher level of apoptosis rate and MDA (P ＜ 0.05),and a lower level of NO systhesis,P-eNOSSer1177/eNOS expression,CAT,SOD,GSH (P ＜ 0.05).Pretreated with tBHQ can reduce the apoptosis rate and MDA level (P ＜ 0.05),improve the amount of NO systhesis,the expression of P-eNOSSer1177/eNOS,the levels of CAT,SOD,and GSH in these cells (P ＜ 0.05).[Conclusion] Activation of Nrf2-ARE pathway can improve endothelial dysfunction in aortic endothelial cells induced by uremic serum,and its mechanism might be related with enhancement of the antioxidant stress.

ObjectiveTo evaluate the applicability of modified formulas based on plasma creatinine levels in Chinese patients with chronic kidney disease (CKD). MethodsA total of 327 CKD patients were investigated. Glomerular filtration rate (GFR) was estimated with Chinese equations and Ruijin equation. The accuracy of estimated GFR was compared with 99mTc-DTPA-GFR (sGFR) in CKD patients. ResultsBland-Ahman analysis demonstrated that Ruijin equation was more consistent with sGFR than the other equations. But all the equations were not well consistent with sGFR. Linear regression showed that the slopes of Ruijin equation and MDRD-1 equation were closer to the identical line. 15%, 30% and 50% accuracy of Ruijin equation were higher than the other equations. But 30% accuracy of Ruijin equation was still less than 70%. When the accuracy of estimated GFRs was compared with sGFR in different stages of CKD, GFR estimated by Ruijin equation showed good results. ConclusionsWhen plasma creatinine is checked with enzymatic method, modified GFR estimation equations may show great bias in Chinese CKD patients. More clinical trails should be carried out to evaluate and identify the application of modified GFR estimation equations in Chinese patients with CKD.

Objective To investigate if Rac1 GTPase activation plays an important role in hyperpermeability and tyresine phosphorylation of tight junction induced by vascular endothelial growth factor (VEGF) in glomertdar endothelial cells (GEnCs). Methods Primary cultured rat endothelial cells were used as experimental model. The effect of VEGF at different concentrations (5 or 50 μg/L) on endothelial permeability was investigated by transendothelial electrical resistance (TEER). The permeability of GEnCs transfected with wild type Rac1 (wtRacl) or dominant negative Racl (N17Rac1) was also detected. Immune precipitation and immune blotting were used to detect the tyrosine phosphor-occludin in GEnCs. Results VEGF at high concentration (50 μg/L) induced hyperpermeability in GEnCs (P<0.05). At the same time, GTP-binding and membrane-bound Racl GTPase significantly increased(P<0.01)in GEnCs. Tyrosine phosphor-occludin was also increased (P<0.05) under VEGF stimulation. However, transfection of GEnCs with N17Rac1 dramatically attenuated the effect of VEGF on tyrosine phospho-occludin and endothelial cell permeability. Conclusions Increased VEGF can induce hyperpermeability in glomerular endothelial cells, which is related to occludin tyrosine phosphorylation through Racl activation. It provides a framework for understanding the role of VEGF-induced Racl-phospho-occludin pathway in the integrity of barrier function in the diabetic milieu.

Objective To investigate the effects of supernatant of cultured mesangial vcells with serum IgA1 from [gA nephropathy patients on apoptosis of podocyte. Methods Jacalin affinity chromatography and Sephacryl S-200 molecular sieve chromatography were used to isolate IgA1. Apoptosis rate of podocyte was assessed by flow cytometer. Monomeric IgA1 (mIgA1) was transformed to aggregated IgA1(aIgA1) by heating. IgA-mesangial cell supernatant was prepared by collecting spent medium in which growth-arrested mesangial cells were incubated with different aIgA1, then the medium with RPMI 1640 containing 0.5%FBS was cultured with growth-arrested podocyte. Real time PCR was used to detect the mRNA expression of Bcl-2, Bax, Fas and Fas-L. Results Apoptosis rate of podocyte by supematant of cultured mesangial cell with algal from IgAN patients was higher than that from healthy and control groups [(28.5±5.9 ) % vs (22.5± 5.8)%, (20.5±4.5)%, all P<0.05]. Fas mRNA expression of podocyte exposed to supematant of cultured mesangial cells with aIgA1 from IgAN patients increased significantly and was 1.89 folds of control (P<0.05), while Bcl-2 mRNA expression significantly decreased and was 72% of control (P<0.05). The concentrations of Ang Ⅱ and TGF-β in supernatant of cultured mesangial cells with IgA1 from IgA nephropathy were significantly higher than those from healthy control [(13.2±3.4) ng/L vs (8.2±2.3) ng/L, /'<0.05; (15.4±3.4) ng/L vs (10.8±3.2) ng/L, P<0.05]. Conclusion Supernatant of cultured mesangial cells with IgA1 from IgA nephroapthy patients can induce apoptosis of podocyte, which may play a role in the progression of IgAN.

Objective To investigate the effect of advanced glycation end products (AGEs) on the disruption of tight junctions in rat glomerular endothelial cells (rGEnCs) and the role of renin-angiotensin system (RAS) in this pathological procedure.Methods Primary cultured rGEnCs were incubated with AGEs at concentrations of 20 mg/L,40 mg/L and 80 mg/L,for 6 h,12 h and 24 h respectively.The cells were treated with captopril (1 mmol/L) or valsartan (10 μ mol/L)to block RAS.The endothelial permeability was investigated by transendothelial electrical resistance and the flux of fluorescein isothiocyanate-conjugated bovine serum albumin.The expression of AGEs receptor (RAGE),tight junction proteins [occludin,claudin-5,junctional adhesion molecules A (JAM-A) and zona occludens-1 (ZO-1)]and RAS components [angiotensinogen,renin and angiotensin Ⅱ type 1 receptor (AT1)]were detected by Western blotting.Immunofluorescence was used to demonstrate the disruptions of the tight junction proteins.The activity of angiotensin converting enzyme (ACE) was evaluated by UV spectrophotometry.Angiotensin Ⅱ (Ang Ⅱ ) was measured by enzyme immunoassay.Results The monolayer permeability,the expression of RAGE,the activity of ACE,the concentration of Ang Ⅱ and the expression of AT1 of rGEnCs were increased after induced by AGEs.Meanwhile,AGEs decreased the expression of occludin,claudin5 and JAM-A and induced disruption of tight junction proteins.Pretreatment with anti-RAGE antibody (100 mg/L),captopril or valsartan could attenuate the detrimental effect of AGEs.Conclusion The changes of permeability induced by AGEs in glomerular endothelial cells are partly mediated by RAS through RAGE.

Objective To explore the effect of high glucose on the renin-angiotensin system (RAS) in rat glomerular endothelial cells and its probable mechanism.Methods Rat glomerular endothelial cells were stimulated by culture medium containing 5 mmol/L or 30 mmol/L glucose (with or without pre-treatment of captopril and chymostatin) for 12 h,24 h,48 h,72 h.ELISA,real-time PCR,Western blotting and confocal immunofluorescence microscopy were employed to examine the follows:the angiotensin Ⅱ (Ang Ⅱ ) concentration in cell lysate and the culture medium; the mRNA levels of angiotensinogen and renin; the protein levels of angiotensin Ⅱtype 1 receptor (AT1R),angiotensin Ⅱ type 2 receptor (AT2R),renin and angiotensinogen in cell lysate; localization of intracellular AT1R,AT2R and renin.Results Exposure to high glucose for only 12 h or 72 h resulted in a significant increase of Ang Ⅱ levels in the culture medium compared with control cells (P＜0.05),but only exposure to high glucose for 72 h resulted in a significant increase of Ang Ⅱ levels in the cell lysate compared with control cells (P＜0.05).However,exposure to high glucose for 24 h or 48 h had no effects on Ang Ⅱ levels in the cell lysate or culture medium.The captopril and chymostatin were able to antagonize high glucose induced Ang Ⅱ generation when exposure time was 72 h but not 12 h.Exposure to high glucoseincreased the mRNA level of angiotensinogen,but reduced the level of renin mRNA,meanwhile angiotensinogen protein increased,and AT1 protein reduced,but protein levels of AT2R and renin were unchanged.However,the transformation of AT2R from the cell nucleus to cytoplasm was observed.Conclusions High glucose can activate the local renin-angiotensin system in rat glomerular endothelial cells,and the probable mechanism may contribute to ACE and non-ACE pathways.The effects of high glucose on glomerular endothelial cells may also involve in the substrate and receptors of Ang Ⅱ.

Objective To evaluate the applicability of glomerular filtration rate (GFR) caJcuated by Cock-croft-Gault equation, MDRD1 equation and abbreviated MDRD equation in the chronic glomerulonephritis. Methods 143 chronic kidney disease(CKD) patients diagnosed in our hospital from January 2005 to December 2007 were investigated. Glomerular filtration rate(GFR) was estimated with Cockcroft-Gault equation, MDRD1 equation and ab-breviated MDRD equation. The accuracy of estimated GFRs was compared with 99Tcm-DTPA-GFR in chronic glomer-ulonephritis patients. Results Bland-Altman analysis demonstrated that Cockcroft-Gault equation was more consist-ent with GFR and sGFR than the other equations. Linear regression analysis showed that the slopes of Cockcroft-Gault equation were closer to the identical line,with a smaller bias and higher accuracy . When compared, the bias as well as accuracy of estimated GFRs with sGFR in different stages of CKD,Cockcroft-Gault equation showed good results. Condusion When plasma creatinine is checked with enzymatic method, Cockcroft-Gault equation is more appropriate than MDRD1 equation and abbreviated MDRD equation for estimating renal function in chronic glomeru-lonephritis patients.