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

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 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 explore the features and influencing factors of ambulatory blood pressure in chronic kidney disease (CKD) patients.Methods A total of 540 CKD patients from May 2010 to May 2012 in our department were enrolled in this study.Ambulatory blood pressure monitoring was carried out.Blood pressure (BP),proteinuria and other clinical parameters were measured regularly.Ultrasonography was used to evaluate cardiac structure and function,carotid intima-media thickness and plaque.Univariate and multivariate analysis were used to examine the association between BP and clinical parameters.Results 63.9％ of CKD patients was non-dipper BP pattern,and 36.1％ was dipper BP pattern.As compared to dipper BP patients,those with non-dipper BP had higher ratio of nighttime/daytime proteinuria (0.51±0.29 vs 0.42±0.21,P ＜ 0.01),lower estimated glomerular filtration rate (eGFR) [(56.2±48.2) vs (75.5±56.5) ml· min 1 · (1.73 m2)-1,P ＜ 0.01],higher serum cystatin C[(2.8±2.0) mg/L vs (2.1±2.0) mg/L,P ＜ 0.01],higher left ventricular mass index [(53.7±23.2) vs (45.1± 16.3) g/m2,P ＜ 0.01] and severely damaged left ventricular diastolic function and higher carotid intima-media thickness [(0.7±0.3) vs (0.6±0.2) mm,P＜ 0.01].Nighttime blood pressure was independent predictor for proteinuria,eGFR and left ventricular mass index.Conclusions Nondipper blood pressure pattern is very common in CKD patients.Nighttime pressure is closely associated to renal damage and cardiovascular injuries.

Objective To analyze the prevalence,risk factors of kidney disease in type 2 diabetic patients with KDIGO classification of chronic kidney disease,also to study cardiovascular and cerebrovascular diseases and death in these patients,so as to investigate the significance of the KDIGO classification system.Methods One thousand six hundred and forty-five type 2 diabetic patients who were in hospitalization from June 2008 to December 2012 were grouped according to the KDIGO classification of chronic kidney disease and the incidence of vascular disease was analyzed based on the classification.Clinical features were compared between patients with or without kidney disease.The risk factors of kidney disease and the death of diabetic patients were also investigated.Results There were 915 male and 730 female,aged a median (57.86±12.54) years with (6.35±6.30) years duration of diabetes mellitus among the 1645 cases,and 37.2％ of patients had concomitant kidney disease.According to the classi fi cation of CKD,patients in CKD group 3a,group 3b and CKD group 4-5 accounted for 5.7％,3.5％ and 7.6％,while 33.4％ of patients had proteinuria,among which 19.5％ with microalbuminuria,13.5％ with macroalbuminuria.On complications,patients with hypertension accounted for 49.5％,hyperlipidemia 67.7％,diabetic retinopathy 27.4％,cardiovascular and cerebrovascular diseases 18.5％ (coronary artery disease 16.5％,cerebrovascular diseases 8.8％).Statistical difference was detected in the incidence of diabetic retinopathy,coronary artery disease and cerebrovascular diseases between CKD group 3a and 3b (P ＜ 0.05).The duration of diabetes,concomitant hypertention especially with elevated systolic blood pressure,diabetic retinopathy and hyperuricemia were the independent risk factors for type 2 diabetic patients with kidney disease.Age,Scr,complicating cardiovascular and cerebrovascular diseases and advanced CKD stage were the independent risk factors for the death of type 2 diabetic patients with kidney disease.Conclusion KDIGO classification of chronic kidney disease enables better staging of kidney diseases in diabetic patients for management and prognosis.Diabetic patients have a higher prevalence of renal diseases and cardiovascular and cerebrovascular events than the general population.Early control of factors such as blood pressure and serum uric acid can delay the progression of kidney disease,and the predictive role of diabetic retinopathy should be emphasized.

Objective To investigate whether high glucose can induce endothelial-mesenchymal transition (EndMT) in glomerular endothelial cells and the role of TGF-β in the process.Methods Rat glomerular endothelial cells were divided into five groups:normal glucose (NG,5.5mmol/L),high glucose (HG,15,30 mmol/L),TGF-β inhibition (HG+ LY36,30 mmol/L glucose + 10 μmol/L LY364947),hyperosmotic control (M,5.5 mmol/L glucose+25.5 mmol/L mannitol) and solvent control (D,5.5 mmol/L glucose + 1 ml/L DMSO).Western blotting was performed to detect relative protein quantities of endothelial marker claudin 5 and mesenchymal marker α-smooth muscle actin (α-SMA).TGF-β1 and TGF-β2 mRNA levels were measured by real-time PCR.Vascular endothelial marker VE-cadherin and mesenchymal marker α-SMA were detected by immunofluorescent stain and observed by confocal microscopy.Results Compared with NG,the expression of claudin5 protein in HG (15 or 30 mmol/L) was up-regulated while expression of α-SMA protein was down-regulated (P ＜0.05).Both TGF-β1 and TGF-β2 mRNA levels increased as well (P ＜ 0.05).However,when compared with HG,the claudin 5 levels increased while α-SMA decreased in TGF-β inhibition group.No significant changes were observed in hyperosmotic or solvent control group.Confocal microscopy showed the transformation of cells from a cobblestone-liked shape to a spindle one,and a decreasing expression of VE-cadherin while an increasing α-SMA in HG group (P ＜ 0.05),whereas TGF-β inhibition partly attenuated those changes in both morphological and protein levels.Conclusions High glucose treatment of glomerular endothelial cells results in an increase in the level of TGF-β1 and TGF-β2 mRNA and leads to endothelial-mesenchymal transiton.Inhibition of TGF-β partly prevents this process,indicating that TGF-β plays a crucial role in high-glucose-induced glomerular endothelial-mesenchymal transiton.