Objective To evaluate the effects of Ang Ⅱ on the expression of IQ domain GTPase-activating protein1 (IQGAP1) and apoptosis of glomerular cells,and to explore the role of IQGAP1 in Ang Ⅱ-induced apoptosis of glomerular cells.Methods Thirty-six male Wistar rats were randomly assigned to receive either saline or Ang Ⅱ by osmotic mini-pump,or be used as normal control.The systolic blood pressure and proteinuria were measured at day 7,14,21,28.After the animals were sacrificed at day 14,28 respectively,the kidneys were collected.Renal pathological change,glomerular cell apoptosis were observed.The expression of glomerular IQGAP1 was assessed by immunohistochemistry,immunofluorescence and Western blotting.The activation of caspase-3 and phosphorylation of extracellular regulated protein kinases 1 and 2 (ERK1/2) were determined by Western blotting.Results Ang Ⅱ-infused rats developed significant hypertension and marked proteinuria.Mild glomerular mesangial cell proliferation and mesangial matrix increase were also observed in Ang Ⅱ-infused rats.The number of apoptotic glomerular cells in Ang Ⅱ-infused rats was significantly more than that in normal control (P ＜ 0.05).The expression of IQGAP1 in glomeruli distributed linearly along the capillary loops.Ang Ⅱ infusion up-regulated the expression of glomerular IQGAP1,which had an significantly positive correlation with activation of caspase-3(r =0.689,P ＜ 0.05) and phosphorylation of ERK1/2 (r =0.658,P ＜ 0.05).Conclusion The enhanced expression of IQGAP1 may be involved in Ang Ⅱ-induced glomerular cells apoptosis via activation of ERK1/2 signaling pathway.

Objective To investigate the correlation of M-type phospholipase A2 receptor (PLA2R) genetic polymorphism in two single nucleotide polymorphisms (SNPs) with idiopathic membranous nephropathy (IMN) of Chinese Han population in Northeast China.Methods A total of 327 individuals were enrolled in the study including 95 adult patients with biopsy-proved IMN (IMN group) followed up for (25.4±11.6) months and 232 healthy people identified by healthy examination in China-Japan Union Hospital of Jilin University (HC group).Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotype and allele frequency of rs35771982 and rs3828323 site in PLA2R gene.The x2 test was performed to compare the distribution difference of allelic frequency and genotype frequency of the two sites in PLA2R gene between two groups.Unconditional Logistic regression analysis was used to determine the risk factor of IMN.Results IMN and HC group were matched in male predominance and body mass index (BMI).Patients with IMN were older than the healthy controls and had higher Scr,serum total cholesterol (TC),24-hour urine protein level and lower serum albumin (Alb) level,lower estimated glomerular filtration rate (eGFR) than the healthy controls (all P ＜ 0.01).The CC genotype frequency and the C allele frequency at SNP rs35771982 site of PLA2R gene in IMN group were significantly higher than those in HC group (x2=13.658,P=0.001;x2=15.315,P=9.10× 10-5),whereas there was no distribution difference of genotype and allele frequency at rs3828323 site between two groups (x2=2.844,P =0.241; x2 =2.959,P=0.085).The CC genotype at rs35771982 site in patients with IMN was not related to,age,gender,BMI,blood pressure and several laboratory indexes such as Alb,TC,Scr,eGFR and 24-hour urine protein level (all P ＞ 0.05).Unconditional Logistic regression analysis revealed that the genotype at rs35771982,age,TC,Scr and eGFR were correlated with IMN occurrence.The CC genotype at rs35771982 was the risk factor of IMN (OR =4.408,95％CI 1.488-13.058).Conclusions The CC genotype and C allele at rs35771982 site in PLA2R may be associated with the susceptibility to IMN,whereas the correlation between gene polymorphism at rs3828323 site and IMN is not demonstrated.The CC genotype at rs35771982 is the independent risk factor of IMN in Chinese Han population in Northeast China.

Objective To evaluate the values of urinary liver-fatty acid binding protein (uL-FABP) and urinary neutrophil gelatinase-associated lipocalin (uNGAL) in diagnosis of acute kidney injury (AKI) caused by obstructive nephropathy and in the prediction of renal prognosis.Methods Clinical data of 30 patients with obstructive nephropathy were collected prospectively.uL-FABP and uNGAL were measured by ELISA at various time points.Risk factors of the renal outcome were evaluated.The patients were followed up for at least one year.Results Patients with AK1 had higher levels of uL-FABP and uNGAL compared to those without AKI [700.00(154.62-1216.14) μg/g· Cr vs 26.90 (16.77-41.38) μg/g·Cr; 1266.69 (671.57-3396.07) μg/g·Cr vs 179.12 (90.98-215.16) μg/g·Cr,all P ＜ 0.01].Positive correlations of uL-FABP and uNGAL with serum creatinine were found (r =0.552,0.553,all P ＜ 0.01).The AUCs of uL-FABP and uNGAL to detect AKI were 0.925 and 0.900.Patients with non complete renal recovery had higher levels of uL-FABP before operation and 72-hour after operation compared to those with complete renal recovery (all P ＜ 0.01).Before operation,the AUC of uL-FABP to detect renal prognosis was 0.948,sensitivity was 85.7％ and specificity was 90.9％.72-hour after operation,the AUC of uL-FABP to detect renal prognosis was 0.935,sensitivity was 85.7％ and specificity was 90.9％.Kaplan-Meier analysis revealed that uL-FABP before operation over 366.57 μg/g · Cr or uL-FABP 72-hour after operation over 223.60 μg/g · Cr were closely related to the poor progression of renal function.Conclusions uL-FABP and uL-NGAL have good accuracy in detecting AKI.The level of uL-FABP before operation and 72-hour after operation is helpful to predict the renal outcome of obstructive nephropathy.

Objective To investingate the effect of low-density lipoprotein (LDL) on epithelial -mesenchymal transition and extracellular matrix (ECM) accumulation in human peritoneal mesothelial cells (HPMCs).Methods (1)HPMCs were randomly divided into control group,LDL group (100 mg/L) and LDL (100 mg/L) + lactoferrin (100 mg/L,LDL receptor blocking agent) group.After co-cultured for 24 h,the expression of LDL receptor in HPMCs was examined by immunofluorescence staining,and the LDL uptake by HPMCs was observed with oil red O staining.(2)HPMCs were cultured with different concentrations of LDL (0,25,50,100 mg/L).After co-cultured for 24 h,the change of cell morphology was observed by inverted phase contrast microscope,and the expression of α-smooth muscle actin (α-SMA) was examined by immunofluorescence.(3) HPMCs were randomly divided into control group (5.6 mmol/L glucose),mannitol group (M,2.18％ mannitol),low glucose group (LG,30 mmol/L),high glucose group (HG,120 mmol/L) and HG + LDL group (120 mmol/L glucose + 100 mg/L LDL).Cocultured for 48 h,the mRNA expression of α-SMA,E-cadherin and type 1 plasminogen activator inhibitor (PAI-1) was detected by real-time quantitative PCR,the protein expression of α-SMA was detected by Western blotting,the content of type I collagen (Col I) and PAI-1 in supernatant was detected by ELISA.Results (1) After co-cultured with LDL for 24 h,the expressin of LDL receptor was found on the cell membrane of HPMCs.Oil red staining showed that LDL could be uptaken into the cells and abolished by LDL receptor blocker.(2) HPMCs tended to be loosely intercellular connected to each ofher,and prsesnted significant formation of fibroblast-like spindle morphology.The cytoplasm immunofluorescence intensity of α-SMA gradually increased with the increase of LDL concentration.Compared to the control group,the expressions of α-SMA mRNA and protein were significantly increased,and the expression of E-cadherin mRNA was decreased in HG + LDL group(all P ＜ 0.05).But the expressions of the parameters above-mentioned were not significant different between HG group and HG + LDL group or between HG group and control group.(3) Compared with HG group or control group,the concentrations of Col Ⅰ [(19.27±0.17) μg/L vs (14.09±0.30) μg/L or (14.81±0.91) μg/L,all P ＜ 0.05] and PAI-1 [(498.24±76.91) ng/L vs (342.19±30.43) ng/L or (220.39±33.82) ng/L,all P ＜ 0.05] in supernatant of HPMCs were significantly up-regulated in HG + LDL group,meanwhile the expression of PAI-1 mRNA was significantly higer than that in control group (P =0.022).Conclusions HPMCs uptake LDL into cells via LDL receptors.LDL can induce HPMCs transdifferentiation in the condition of high glucose,increase the secretion of Col Ⅰ,inhibit the degradation of ECM through up-regulating the expression of PAI-1,and lead to ECM accumulation.

Objective To investigate the effect of hepatitis B virus X (HBX) gene on apoptosis and immune molecules of human proximal renal tubular epithelial cell line (HK-2).Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.Untransfected HK-2 cells and those transfected with empty vector were used as controls.The TLR4 expression was detected by real-time PCR and Western blotting.The apoptosis of cells and expression of MHC-Ⅱ and CD40 were detected by flow cytometry,and the contents of IL-4 and IFN-γ in the supernatant were detected by EIISA.Results Compared with control groups,the number of apoptotic cells was significantly increased in the HBX transfection group (P ＜ 0.05),and the expressions of TLR4,MHC-Ⅱ and CD40 were also significantly increased in the HBX transfection group (all P＜0.05).IFN-γ level in the supernatant of HBX transfection group was higher (P ＜ 0.05),but IL-4 level was lower as compared to control groups (all P ＜ 0.05).Conclusions Over-expression of HBX gene may induce apoptosis of HK-2 cells and upregulate the expression of immune molecules of renal tubular epithelial cells leading to injury of cells and dysfunction of immunomicroenviroment.

Objective To identify the prevalence of different amyloid types in renal biopsies.Methods The renal biopsies of 205 patients diagnosed as renal amyloidosis from January 1990 to December 2011 were reassessed.Immunohistochemistry was performed with a penal of antibodies directed against λ-light chain,κ-light chain,amyloid A,fibrinogen,transthyretin,apolipoprotein A1,and lysozyme.Immune electron microscopy and gene analysis were performed when the results of immunohistochemistry were indeterminate.Results Among 205 patients,190 cases (92.7％) were classified as immunoglobulin light chain amyloidosis (AL),1 (0.5％) case as amyloid A amyloidosis and 1 (0.5％) case as fibrinogen A α-chain amyloidosis.The amyloid types of remaining 13 (6.3％) cases were undetermined.In the AL patients,the distribution of λ and κ was 6.6∶1.Conclusion AL is the most common form of renal amyloidosis in China,with a predominant light chain type of λ.

Objective To compare the outcomes of patients starting peritoneal dialysis (PD)within two weeks and more than two weeks after catheter implantation.Methods All the patients undergoing Tenckhoff catheter implantation and initiating PD in Renji Hospital from January 2001 to December 2010 were enrolled in the study.Patients started PD within 2 weeks after catheter insertion were defined as urgent group,and those started PD 2 weeks later were defined as planned group.Kaplan-Meier curves and Log-rank tests were used to compare outcomes between two groups.Results Among 657 patients in this study,median break-in period was 6 days of 469 (71.4％)patients in urgent group and 26 days of 188 (28.6％) patients in planned group.Compared to planned group,patients of urgent group were younger [(52.6± 17.3) vs (56.1± 15.3) year,P =0.017],had less eGFR [(5.36±2.03) vs (6.50±2.50) ml· min-1 · (1.73 m2)-1,P ＜ 0.01],lower serum albumin [(34.0±5.7) vs (36.2±5.9) g/L,P ＜ 0.01] and hemoglobin [(76.9± 18.8) vs (80.8 ± 17.9) g/L,P =0.018],and higher phosphate [(2.19±0.67) vs (1.98±0.52) mmol/L,P＜ 0.01].Urgent group presented more catheter dysfunctions needed to transfer to hemodialysis (2.1％ vs 0％,P =0.044).The 1-,2-,3-and 5-year technique survival rates of urgent and planned group were 94％ vs 98％,92％ vs 94％,90％ vs 92％and 86％ vs 85％ respectively.There was no significant difference in technique survival (Log-rank =1.536,P =0.22) and peritonitis-free survival (Log-rank =0.035,P =0.85) between two groups.The 1-,2-,3-and 5-year patient survival rates of urgent and planned group were 90％ vs 95％,81％ vs 90％,74％ vs 79％ and 67％ vs 74％ respectively,and no significant difference was found (Log-rank =2.364,P =0.12).Conclusions Although patients needing urgent initial PD have poorer residual renal function and nutritional condition compared to those of planned initial PD,their outcomes are similar.Peritoneal dialysis may be a feasible and safe dialysis modality for patients who need urgent start.

Objective To investigate the long-term prognostic factors and the significance of serum cardiac troponin T (cTnT) and C-reactive protein (CRP) in maintenance hemodialysis (MHD) patients.Methods Clinical data of 76 MHD patients in our hospital from January 2002 to January 2003 were retrospectively analyzed.Time and cause of death in the next 10 years were recorded.Survival rate was calculated by Kaplan-Meier and impact factors of long-term prognosis were explored.Significance of cTnT and CRP was elucidated by COX regression analysis.Results CRP was positive in 28 cases (36.8％) and cTnT was positive in 22 cases (28.9％) among 76 patients.The median survival time was 37.9 months,2-year survival rate was 65.9％ and 10-year survival rate was 24.2％.Univariate analysis found positive CRP,positive cTnT,old age,diabetes,cardiocerebrovascular disease,anemia,low serum albumin,Kt/V decline were associated with long-term prognosis.Multivariate analysis showed that increased age (P =0.010),cardiocerebrovascular disease (P =0.048),positive cTnT (P =0.036),positive CRP (P =0.009) were independent risk factors of the 10-year survival of MHD patients.Ten-year mortality of cardiocerebrovascular diseases in positive cTnT group was not significantly different as compared with negative cTnT group (50.0％ vs 35.4％,P =0.248).But the positive cTnT group had higher 2-year mortality than negative cTnT group (40.9％ vs 14.6％,P =0.015).Mortality of cardiocerebrovascular disease was higher in positive CRP group as compared to negative CRP group at both 2-year and 10-year time (48.1％ vs 7.0％,P =0.000; 66.7％ vs 23.3％,P =0.000).Compared with both negative cTnT and CRP group,both positive cTnT and CRP group had much higher all-cause mortality (92.9％ vs 55.6％,P =0.030),higher mortality of cardiocerebrovascular disease at 10-year (64.3％ vs 25.0％,P =0.009),and higher mortality of cardiocerebrovascular disease at 2-year (57.1％ vs 5.6％,P =0.000).Conclusions Aging,cardiocerebrovascular disease,positive cTnT and positive CRP are independent risk factors of long-term prognosis for MHD patients.Positive cTnT can predict cardiocerebrovascular mortality of MHD patients in 2 years,while positive CRP can predict short-and long-term cardiocerebrovascular mortality.Positive cTnT combined with positive CRP may be more valuable in predicting the poor prognosis of MHD patients.

Objective To compared two classical rat models of nephrotic syndrome and to provide some reference data to researchers.Methods Thirty male SD rats were randomly divided into control group,puromycin aminonucleoside-induced nephrotic syndrome (PAN) group and adriamycininduced nephrotic syndrome (ADR) group.The body weight,twenty four hour proteinuria level,serum albumin concentration,cholesterol concentration,creatinine and urea concentration were measured.The renal pathology change was evaluated.The drug toxic effects,administration methods and the costs were also compared.Results There was no significant difference in body weight and hair color between control group and PAN group.Compared to control group,the body weight of the rats significantly decreased at day 15 and day 21 in ADR group (P ＜ 0.01),accompanied by epilation and diarrhea.Compared to control group,the 24-hour urinary protein levels increased significantly at day 10 (P ＜ 0.01),day 15 (P ＜ 0.01),and reached the peak level at day 15 (P ＜ 0.01),day 21 (P ＜ 0.01) in PAN group and ADR group respectively.Compared to control group,the serum albumin concentration decreased significantly at day 10 (P＜0.01),and return to normal level at day 15.The serum cholesterol concentration was increased significantly at day 10 (P ＜ 0.01) and return to normal at day 15 in PAN group.Compared to control group,the serum albumin concentration was decreased significantly at day 15 (P＜0.05) and return to normal at day 21 in ADR group.No significant difference of serum creatinine and serum urea nitrogen levels were found among three groups.Compared to control group,the width of foot process increased significantly at day10 (P ＜ 0.01) and day 15 (P ＜ 0.05) in PAN group and ADR group respectively.To successfully induce a nephrotic rat model (per 100 g),the cost of PAN group was 3.1 times of ADR group (578.10 yuan vs 186.94 yuan).Conclusions Nephrotic syndrome can be induced by both PAN and ADR.The administration of PAN via intraperitoneal injection is more convenient as compared to ADR via tail intravenous injection.Compared to ADR,PAN can induce nephrotic syndrome model more rapidly,with more consistent detection index,and less toxic effects,but its cost is more expensive.

Objective To investigate the correlation between glycosylated hemoglobin (HbAlc) and carotid intima-media thickness (CIMT) in non-diabetic peritoneal dialysis patients.Methods Forty-two non-diabetic peritoneal dialysis adult patients were enrolled in this study [mean age was (48.2± 12.3) years,50％ was male].CIMT was determined by carotid ultrasound.Patients were divided into two groups according to CIMT:CIMT normal group (CIMT ＜ 0.9 mm) and CIMT thickening group (CIMT≥0.9 mm).HbAlc,2-hour postprandial blood glucose (2hPBG) and other factors of the patients were analyzed with Spearman rank correlation and multiple linear regression.Results CIMT was correlated with age,2hPBG,LDL-C,TG,TC,HbA1c in non-diabetic peritoneal dialysis patients (r =0.355,0.373,0.416,0.345,0.351,0.456,all P ＜ 0.05).Multiple linear regression showed that HbA1c was the most powerful influence factor of CIMT(β =0.459).Conclusion HbA1c level is positively correlated with CIMT and may be a predictor of carotid atherosclerosis in non-diabetic peritoneal dialysis patients.