Objective To elucidate the inheritance and clinical characteristics, and to analyze the SLC4A1 gene mutation in a family with autosomal dominant distal renal tubular acidosis (dRTA). Methods Family investigation and renal tubular acidification examination were performed. SLC4A1 gene mutation was analyzed by direct sequencing analysis in this family. One hundred unrelated healthy subjects were also analyzed as controls. Results Six patients were diagnosed as autosomal dominant dRTA with severe phenotype in this family. All the patients were heterozygous for a novel 1-bp duplication in exon 20 of SLC4A1 (c.2713dupG, band 3 Qingdao). This duplication resulted in a reading frame for 15 extra condons (instead of 8) before the new stop condon at position 919 （p.Asp905Glyfs15）. Conclusions A novel mutation is identified in association with autosomal dominant dRTA with severe phenotype. This is the first report of hereditary dRTA in China.

Objective To perform the exclusive gene mapping around four known genes associated with familial focal segmental glomerulosclerosis（FFSGS）in a FFSGS kindred using linkage analysis． Methods The clinical features of available members in this kindred were investigated． Nine microsatellite markers around WT1, TRPC6, CD2AP and NPHS2 loci were selected and applied to analyze the linkage in the available family members. By two-point linkage analysis and allele sharing analysis, exclusive gene mapping on NPHS2, WT1, TRPC6 and CD2AP were carried out in this FFSGS kindred． Results Autosomal dominant inheritance pattern was demonstrated in phenotypes of this family. Of 19 family members, 1 suffered from end-stage renal disease (ESRD), 4 individuals were found to have proteinuria, and 2 affected individuals underwent renal biopsy and were diagnosed as FSGS. The youngest age of onset was 10 years old. This autosomal dominant FSGS was evaluated for linkage using microsatellite markers including D1S196, D1S218, D1S238, D11S935, D11S898, D11S908, D11S1986, D6S936, D6S1566 and so on. The maximal two-point logarithm of odds(LOD) score was 0.32 at θ=0 in marker D11S1986. These markers were not cosegregated with the disease loci in all the affected members, which indicated that there was no linkage between these markers and four FFSGS related genes (CD2AP, WT1, NPHS2 and TRPC6) in the kindred. Conclusions The inheritance pattern of this kindred is autosomal dominant. NPHS2, WT1, TRPC6 and CD2AP are not the disease-causing genes for this family.

Objective To compare survival in patients on hemodialysis (HD) and peritoneal dialysis (PD) and to explore the main risk factors affecting survival of dialysis. Methods The data of initial dialysis patients of 18 years or older between 1 January 2005 and 31 December 2008 were analyzed retrospectively. All the patients were followed up until 31 March 2009. Survival analysis was performed using Kaplan-Meier method, log-rank test and Cox regression model. Results A total of 460 dialysis patients were enrolled, among whom 247 patients received HD therapy and 213 patients received PD therapy. There was no significant difference of age, body mass index (BMI), estimated glomerular filtration rate (eGFR), mean arterial pressure (MAP) at the beginning of dialysis and the history of cerebrovascular accident, Charlson comorbidity index (CCI) before dialysis between patients on HD and PD (all P＞0.05). The median follow-up time was 17.9 (0.25-51) months. By analysis of treatment intention, Kaplan-Meier survival curve revealed that overall mortality rate was significantly lower in HD group (P＜0.05, log-rank test). Mortality of two groups was similar in the first 1 year (P=0.14, log-rank test). In the period beyond 1 year, there was a significantly higher mortality in PD group. Sub-group analysis revealed that survival was better in non-diabetic nephropathy (non-DN) patients older than 65 years on HD (P＜0.05). After the adjustment of case-mix differences, the Cox regression model indicated that there was no significant difference of survival between HD and PD patients [HR (HD:PD)=0.778, 95%CI 0.483-1.254, P=0.303], while age (HR=1.051, 95%CI 1.030- 1.073, P＜0.01), history of cerebrovascular accident before dialysis (HR=2.032, 95%CI 1.125-3.670, P＜0.05), CCI≥5 before dialysis (HR= 2.592,95%CI 1.230-5.465, P＜0.05), prealbumin (HR=0.022, 95%CI 0.001-0.768, P＜0.05) were the main risk factors of survival in dialysis. Conclusions Survival is similar between HD and PD patients in the first 1 year of dialysis. In the period beyond 1 year, mortality of PD patients is significantly higher. Survival is better in non-DN patients older than 65 years. Age, history of cerebrovascular accident and CCI≥5 before dialysis may be the main risk factors of survival in dialysis.

Objective To investigate the influence of blood glucose level on the estimated glomerular filtration rate (eGFR) by Cockcroft-Gault (CG) and Modification of Diet in Renal Disease (MDRD) formula in diabetic patients, and to explore the difference between CG and MDRD formula to estimate GFR in different levels of blood glucose in the diagnosis of moderate renal insufficiency. Methods A total of 1210 diabetic patients (650 males and 560 females) were enrolled in this study. HbA1c, Scr and isotopic GFR (iGFR) (99m Tc-DTPA) were measured. CG and MDRD formula were used to estimate the GFR (eGFRCG, eGFRMDRD). The patients were divided into normal GFR group [NGFR, n=589, iGFR≥90 ml&#8226;min-1&#8226;(1.73 m2)-1], gently decreased GFR group [GGFR, n=470, 60≤iGFR<90 ml&#8226;min-1&#8226;(1.73 m2)-1], moderate and sever decreased GFR group [MGFR, n=151, 30≤iGFR<60 ml&#8226;min-1&#8226;(1.73 m2)-1] based on the K/DOQI suggestion. According to the quartile HbA1c level (7.1%, 10.5%), patients were divided into four groups. Patients with HbA1c<7.1% were defined as well-controlled group, with HbA1c≥10.5% as poorly- controlled group. Spearman correlation, t test, Bland-Altman analysis and ROC curve were applied to investigate the bias and accuracy of formula, and the influence of blood glucose on eGFR. Results HbA1c was correlated with iGFR (r=0.17, P<0.01), eGFRCG (r=0.22, P<0.01), eGFRMDRD (r=0.29, P<0.01). The correlation between eGFRMDRD and HbA1c was the strongest. eGFRMDRD seemed to overestimate GFR. However, eGFRCG seemed to underestimate GFR in well-controlled group. Bland-Altman analysis indicated that the bias of eGFRMDRD with iGFR in poorly-controlled group was higher than that in well-controlled group. Compared with poorly-controlled group, the 15% and 30% accuracies of eGFRMDRD in well-controlled group were significantly higher. There was no significant difference between poorly-controlled group and well-controlled group in eGFRCG. The bias of eGFRCG with iGFR was significantly higher than that of eGFRMDRD with iGFR. The area under ROC curve was significantly greater in eGFRMDRD than that in eGFRCG (P<0.05) in poorly-controlled group. Conclusions eGFR by MDRD and CG formula in diabetic patients may lead to discrepancies with GFR value. eGFRMDRD overestimates the GFR value. MDRD equation is more accurate and robust in poorly-controlled diabetic patients with moderate renal insufficiency.

Objective To explore the risk factors of acute kidney injury (AKI) in patients with brain trauma and to investigate the role of mannitol. Methods A random cohort of traumatic brain injury patients who were admitted to Neurosurgical Emergency Center of Huashan Hospital, Fudan University from January 2006 to December 2008 was studied. AKI was determined using RIFLE staging criteria for changes in serum creatinine (Scr). By means of Logistic regression analysis, the risk factors of AKI were investigated, and a predictive model was established. Discrimination of the model was assessed using the receiver operating characteristic (ROC) curve. Association between single independent risk factor and AKI was depicted with ROC curve. Adjustment for selection bias was further assessed using propensity score match (PSM) to evaluate the role of mannitol in the development of AKI. Results A total of selected 171 patients were enrolled in the study, including 53 patients in AKI group and 118 patients without AKI as control group. The average age of these 171 patients was (45.92±16.50) years old. Univariate analysis revealed that age, hypertension, emergent surgery, systemic inflammatory response syndrome (SIRS), Glasgow coma score (GCS), sequential organ failure assessment (SOFA) score, respiration component of the SOFA score, coagulation component of the SOFA score, cardiovascular component of the SOFA score, mechanical ventilation time, red blood cell transfusion, plasma transfusion, accumulative dose of furosemide, accumulative dose of torasemide and accumulative dose of mannitol were significantly associated to AKI in patients with brain trauma. Logistic multivariate regression analysis showed that SOFA score (OR=1.516, 95%CI 1.222-1.881, P<0.01), accumulative dose of torasemide (OR=0.016, 95%CI 1.002-1.031, P=0.016) and accumulative dose of mannitol (OR=2.687, 95%CI 1.062-6.800, P=0.037) were independent risk factors of AKI. This model had a good discrimination for AKI with an area under the ROC curve of 0.901 (P<0.01). The accumulative dose of mannitol was identified as a risk factor of AKI by PSM test. Conclusions AKI is a common complication in patients with traumatic brain injury. SOFA score, accumulative dose of torasemide and accumulative dose of mannitol are independent risk factors of AKI following traumatic brain injury.

Objective To analyze the characteristics and relevant factors of anemia in long-term renal transplant recipients. Methods Two hundred and fifty eight long-term renal transplant recipients were enrolled in the study. The incidence and character of anemia was investigated. The correlation of anemia with EPO, tubular function, renal function, reject reaction, immunosuppressive drugs and` cardiocerebrovascular complications was also analyzed. Results In above 258 recipients, the incidence of anemia was 41.1％. Most cases were normocyte and normochromic, some cases were microcytic hypochromic, and some cases were hemolytic anemia. In these anemic recipients, most cases were short of EPO, and some were EPO-resistance. The hemoglobin (Hb) level in anemic recipients was positively correlated with eGFR (r=0.348, P＜0.01) and Ccr (r=0.351, P＜0.01), while was negatively correlated with NAG (r=-0.327, P＜0.01). When renal function was normal, tubulointerstitial damage was more severe in recipients with anemia as compared to those without anemia. The incidences of acute and chronic rejection were higher in recipients with anemia as compared to those without anemia (P＜0.01). The incidence of anemia in CsA+Aza+Pred treatment was 69.0%; in CsA+MMF+Pred treatment was 35.8％; in FK506+MMF+Pred treatment was 34.8％; in SRL+MMF+Pred treatment was 41.7％, respectively. Marrow suppression was the commom adverse reaction of Aza. The Hb level in anemic recipients was negatively correlated with using time of Aza (r=-0.354, P＜0.01); was negatively correlated with dose and using time of MMF (MMF 2-3 g/d, r=-0.285, P＜0.05; r=-0.372, P＜0.01); was negatively correlated with dose and using time of SRL (SRL 2-5 mg/d, r=-0.278, P＜0.05; r=-0.359, P＜0.01). The incidence of cardiocerebrovascular complication was higher in recipients with anemia as compared to those without anemia (P＜0.01). Conclusions The incidence of anemia in long-term renal transplant recipients is quite high, which is not only correlated with tubular function and renal function, but also with EPO, reject reaction and immunosuppressive drugs. Anemia is a risk factor of cardiocerebrovascular complications in renal transplant recipients.

Objective To investigate the executive function (planning and inhibition) in maintenance hemodialysis (MHD) patients. Methods Neuropsychological research methods, including trail making test (A and B), Go-No/Go test and Stroop color naming task, were applied to examine 40 MHD patients and 40 healthy people (NC group). The mean age and education level of two groups were not significantly different. Results MHD group had significantly lower performance of trail making test A and B than NC group (tA=6.52, P<0.01; tB=5.27, P<0.01). MHD group had less scores on baseline condition of Go-No/Go test (t=3.27, P<0.01) and Stroop color naming test (t=3.30, P<0.01) than NC group. In Stroop color naming task test, NC group showed Stroop effect, significant repeated distraction promotion effect and significant negative priming effect, while MHD group showed only Stroop effect. There was significant difference in Stroop effect between MHD group and NC group (t=2.41, P<0.05). Conclusion Cognitive planning ability and inhibition ability of MHD patients are impaired. This pathological change of executive function in MHD patients is independent of normal aging process.

Objective To investigate the effect of fibrocytes and chemokine recepter 7(CCR7) on the process of renal fibrosis induced by adriamycin in rats. Methods Male Sprague-Dawley (SD) 8 weeks old rats (n=52) were divided into two groups randomly, normal control group (CG, n= 20) and model group (MG, n=32). MG rats were injected with 4 mg/kg of adriamycin via tail vein on the first day of the 1st and 2nd week, respectively, while CG rats received 0.9% sodium chloride solution injection of equal volume at the same time. The urine and serum samples were measured at the end of 4, 7, 10 and 13 weeks after the first administration of adriamycin to observe biochemical changes, while the renal tissue of rats was obtained to evaluate renal histological and ultrastructural changes by glomerulosclerosis index and tubulointerstitial index, and to analyze the expression of CD45, collagen I and CCR7 by immunohistochemistry. Results Compared with CG, levels of 24 h urine protein excretion rate (mg, 568±102 vs 21±6, P＜0.01), serum cholesterol (CHO, mmol/L, 6.79±1.47 vs 1.29±0.27, P＜0.01) and triglyceride (TG, mmol/L, 2.57±1.19 vs 0.76±0.11, P＜0.01) increased significantly, whereas serum albumin (Alb, g/L, 21.41±2.11 vs 34.64±0.96, P＜0.01）decreased significantly in the 7th week in MG. The proliferation of mesangial cells, the accumulation of the extracellular matrix and atrophyin or extension of renal tubules in MG were more remarkable than those in CG. There were some cells which expressed CD45 and collagen I simultaneously in the serial sections of renal tissue and renal cortical interstitium in MG. Interstitial infiltration of CD45+ or CCR7+ cells appeared in the 4th week, reached a maximum in the 7th week (P<0.01) and gradually decreased afterward. Conclusions Fibrocytes may be involved in the earlier stage of kidney fibrosis. The immigration of this population from peripheral blood to renal lesion site is possible to be associated with CCR7 protein.

Objective To elucidate the role of oxidative stress-dependent epidermal growth factor receptor (EGFR) trans-activation in the aldosterone (ALDO)-induced phosphatidyl inositol 3-kinase (PI3K)-Akt activation and mesangial cells (MCs) proliferation. Methods The incorporation of 3H-thymidine (3H-TdR) and cell count were used to measure MCs proliferation. Reactive oxygen species (ROS) production was determined by DCFDA fluorescence. EGFR, PI3K, Akt, mammalian target of rapamycin (mTOR), and p70S6K1 phosphorylation were assayed by Western blotting. Results (1) ALDO induced ROS production in time- and dose-dependent manner in cultured human MCs. ROS generation was significantly increased by incubation with 100 nmol/L ALDO for 3 min, and peaked at 60 min. ROS production was increased by 1.50-, 1.70-, and 2.14-fold after stimulation by 100 nmol/L ALDO for 60 min. Antioxidant N-acetylcysteine (NAC) almost completely blocked ALDO-induced MCs proliferation. (2) ALDO induced EGFR phosphorylation in time- and dose-dependent manner in cultured human MCs. EGFR phosphorylation was significantly increased by incubation with 100 nmol/L ALDO for 5 min, and peaked at 60 min. EGFR trans-activation was increased by 2.29-, 3.55-, and 5.25-fold after stimulation by 100 nmol/L ALDO for 60 min. NAC and mineralocorticoid receptor (MR) antagonist eplerenone significantly inhibited ALDO-induced EGFR phosphorylation, and EGFR antagonist AG1478 blocked ALDO-induced MCs proliferation. (3) ALDO significantly stimulated the phosphorylation of PI3K, Akt, mTOR and p70S6K1 by 4.35-, 5.38-, 3.85-, and 3.57-fold. PI3K inhibitor LY 294002, Akt inhibitor, and mTOR inhibitor rapamycin reduced ALDO-induced MCs proliferation by 50%-55%. Conclusion Oxidative stress-dependent EGFR trans-activation mediates ALDO-induced PI3K-Akt-mTOR-p70S6K1 signal pathway activation and MCs proliferation.

Objective To establish Gpx1-Klk1 combined transgenic mouse model, and to investigate the protective effect of Gpx1-Klk1 transgene on renal ischemia reperfusion injury in vivo. Methods pKsp-Gpx1-IRES-Klk1 recombinant plasmid was constructed and purified for microinjection into the male pronucleus of the zygote in order to prepare the Gpx1-Klk1 combined transgenic mice. A controlled thread hanging method was used to prepare the ischemia and reperfusion model in those successful transgenic mice. The C57BL wild-type mice were chosen in the control group. BUN and creatinine level in serum, SOD, MDA, tNOS and iNOS level in renal tissue, and the structure changes of renal tissue were all investigated before and after the experimental ischemia and reperfusion. Results A 6.585 kb-longed pKsp-Gpx1-IRES-Klk1 recombinant plasmid was obtained and identified. A total of 525 gene-injected ovums were transplanted into oviducts of 21 female mice, resulted in 17 pregnant (81.0%). Ten of those 109 born mice were accepted as transgenic positive by PCR assessment (9.2%). Gpx1-Klk1 combined transgenic mice were successfully established, and proved to have strong expression of Gpx1 and Klk1 protein confirmed by Western blotting. The difference between transgenic group and control group was examined. BUN and Scr of transgenic group were obviously lower than those of control (P<0.01). SOD of renal tissue in transgenic group was higher than that of control (P<0.01), while MDA was lower than control (P<0.01). tNOS of renal tissue in both groups increased significantly compared to that of pre-operation, and tNOS of the transgenic group was obviously higher than that of control (P=0.025), while the tissue iNOS of the transgenic group was obviously lower than that of control (P<0.01). After ischemia reperfusion, slight edema was found in renal interstitial tissue of the transgenic group, while relatively few necrosis was seen in renal tubular epithelial cells. The semiquantitative analysis of the tissue injury severity indicated that the injury of transgenic group was less than that of control group (1.58±1.05 vs 3.95±0.80, P<0.05). Conclusions A new type of renal ischemia and reperfusion mice model based on the Gpx1-Klk1 combined transgenic mice is successfully established. Gpx1-Klk1 gene over-expression has obviously protective effect on renal ischemia and reperfusion injury.

Objective To examine the role of aldosterone and WNK4 in redistribution of epithelial Na(+) channel(γENaC) in response to a high-salt diet. Methods 200-250 g weight Sprague-Dawley rats were randomly assigned to five groups: normal salt diet (N, 0.4% NaCl), high salt diet (H, 4% NaCl), low salt diet (L, 0.07% NaCl), H treated with aldosterone (H+A, 4% NaCl with osmotic mini-pump at 1 mg&#8226;kg-1&#8226;d-1 aldosterone), and L treated with spironolactone (L+S, 0.07% NaCl with 0.1 g&#8226;kg-1&#8226;d-1 spironolactone). Results The high salt diet decreased γENaC protein and the low salt diet increased γENaC protein. However, the γENaC of H+A group was higher compared with that of H group and the γENaC of L+S group was lower compared with that of L group. Radioimmunoassay demonstrated that the concentration of plasma aldosterone was decreased by high salt diet compared with that by normal salt diet. On the contrary to the aldosterone, the expression of WNK4 was remarkable enhanced in H group and decreased in L group, but it was decreased in H+A group and enhanced in L+S group. Conclusions The decrease of γENaC during high salt diet is regulated by aldosterone and involves WNK4 activity as well. WNK4 kinase is affected negatively by aldosterone in kidney.

Objective To establish a pure and convenient method for the enrichment of the renal proximal tubular cells(PTCs). Methods The both kidneys of Wistar rats were underlying hemospasia via cardiac apex substituted in situ kidney perfusion before taken out. The renal cortex was separated and sliced into small pieces which were digested in typeⅠ collogen and resuspended in 45% Percoll gradient, then centrifuged by continuous density gradient. Cells were cultured and passaged in DMEM-F12 supplemented with 10% fetal bovine serum. Bred cells were identified by morphology, immunocytochemistry staining of cytokeratin-18 (CK18) and aquaporin 1(AQP1), and alkaline phosphatase staining. Results The tubular fragments obtained from the modified method had very little miscellany of renal glomerulus. After 4-5 days, the cultured cells reached confluence, displaying the typical cobblestone appearance. All the bred cells almost were positive expression in immunocytochemistry staining of CK18 and AQP1, and alkaline phosphatase staining. So they were validated as rat PTCs. Conclusion This modified method can yield large quantity of pure PTCs simply.