Objective To study the pathologic pattern and clinical feature of type 2 diabetes mellitus complicated with chronic kidney diseases (CKD). Methods Clinicopathological features of 155 type 2 diabetic patients complicated with CKD were collected and analyzed retrospectively. The patients were divided into four groups: typical diabetic glomerulopathy (DG), atypical diabetes-related renal disease (ADRD), non-diabetic renal diseases (NDRD) and DG complicated with NDRD. Results Renal biopsies revealed DG accounted for 18.7% of the patients, ADRD accounted for 12.9%, NDRD accounted for 60.0%, and DG complicated with NDRD accounted for 8.4%. In DG group, duration of type 2 diabetes was longer; the level of fast blood glucose, systolic blood pressure, mean arterial pressure and prevalence of diabetic retinopathy (DR) were higher; proteinuria was heavier and evaluated glomerular filtration rate (eGFR) was lower. In ADRD group, body mass index and prevalence of obesity were higher; dyslipidemia was more severe. Gross hematuria and acute renal insufficiency could be only found in NDRD group. Without DR, duration of diabetes under 5 years, gross hematuria, acute renal insufficiency, evidences of autoimmune diseases and proteinuria≥3.5 g/24 h but eGFR≥60 ml/min were specific valuable predictors for NDRD. Conclusions Renal injuries in type 2 diabetic patients are structural heterogeneous, in which NDRD is more common and is different from ADRD and DG. Renal biopsy should be considered when type 2 diabetic patients complicated with CKD present at least one characteristic as follows: duration of diabetes under 5 years, without DR, history of gross hematuria, acute decrease of renal function, evidences of autoimmune diseases and proteinuria≥3.5 g/24 h but eGFR≥ 60 ml/min.

Objective To assess sleep quality and daytime sleepiness in patients on maintenance high flux hemodialysis, and discussed the associated factors. Methods A total of 112 high flux hemodialysis patients and 53 normal subjects were estimated by Pittsburgh sleep quality index (PSQI) and Epworth Sleep Scale (ESS) to assess the sleep quality and day time sleepiness. Global score of these questionnaires were analyzed. Seven components’ scores and 9 reasons for sleep disturbances were compared between “good” (global PSQI≤5) and “bad” (global PSQI>5) sleepers. Sleep quality was compared among different shifts of hemodialysis. The impact of clinical factors on sleep quality were analyzed by multivariate linear regression and logistic regression. Results Compared with control group, hemodialysis group had a higher PSQI (7.02±4.94 vs 3.28±2.79, P<0.05) and a lower ESS score [3(0-6) vs 8(4.25-11.75), P<0.05] . 58% patients were “bad” sleepers and sleep latency was longer (30 min vs 15 min, P<0.05). Insomnia was the main problem. Patients on morning shift, afternoon shift and night shift had similar subjective sleep quality. Age(OR=1.75, P=0.003), dialysis vintage (OR=1.26, P=0.008), hemoglobin (OR=0.64, P=0.008), calcium phosphate product (OR=1.60, P=0.02) were significantly related to sleep quality score. Conclusions Sleep disturbance is common in hemodialysis patients. Older age, longer dialysis vintage, anemia and higher calcium phosphate product are risk factors for poor sleep quality.

Objective To clarify the relationship between clinical manifestation and pathological features of IgA nephropathy (IgAN) patients with mild proteinuria and/or hematuria. Methods Clinicopathological data from 316 biopsy-proven IgAN cases (proteinuria<1 g/24 h and/or hematuria, and Scr<133 μmol/L) from our hospital between January 1993 and October 2009 were studied retrospectively. The renal histopathology was quantified according to Lee’s grading and Katafuchi’s semi-quantitative standard, and the risk factors for renal pathological lesions were evaluated using multifactor logistic regression analysis. Results Among these 316 patients, 123 were male and 193 patients were female. The mean age at the time of renal biopsy was (33.10±10.69) years old. Clinical features were found as follows: hematuria with proteinuria was found in 267 patients (84.5%), isolated hematuria in 24 patients (7.6%), and isolated proteinuria in 25 patients (7.9%). 16.5% of patients had hypertension. The percentages of CKD stage Ⅰ, Ⅱ, Ⅲ were 76.9%, 20.9% and 2.2%, respectively. 31.3% of patients presented Lee’s grade Ⅲ or more severe. 52.8% of patients had various degrees of glomerulosclerosis. Crescent formation was observed in 20.3% of patients. 22.5% of patients showed tubular atrophy; 16.8% showed interstitial fibrosis and 24.7% also had renal vascular lesions. The extent of glomerulosclerosis was negatively correlated with eGFR levels, but positively correlated with the amount of proteinuria and mean arterial pressure (MAP) level (P<0.05). The score of tubulointerstitial lesion was positively correlated with the amount of proteinuria and negatively correlated with eGFR and hemoglobin (Hb) level (P<0.05). The degree of renal vascular lesion was also correlated to MAP level positively and eGFR level negatively (P<0.05). Multifactor logistic regression analysis revealed that proteinuria, Scr and Hb at the time of renal biopsy were independent risk factors for severe renal pathological lesions (Lee’s grade Ⅲ or more severe) with odds ratio of 8.564, 1.031 and 0.975 respectively (all P<0.01). Conclusions Severe renal histological lesions and decrease of renal function may be seen in some IgAN patients with mild proteinuria and/or hematuria. The levels of proteinuria, Scr and Hb are the independent risk factors for severe renal pathological lesions. Renal biopsy is important in these patients in order to make diagnosis and individual treatment.

Objective To investigate the association between podocytes and proteinuria in children with Alport syndrome. Methods Twenty-one children including 13 boys and 8 girls with Alport syndrome were divided into 3 groups according to 24-hour urinary protein, <30 mg/kg as the mild proteinuria group (10 patients), 30 to 50 mg/kg as the moderate proteinuria group (4 patients) and >50 mg/kg as the heavy proteinuria group (7 patients). The correlation between foot process width and the degree of proteinuria was analyzed. By immunoperoxidase staining on renal tissue, the key slit diaphragm molecules nephrin, podocin and podocyte cytoskeleton-associated molecule synaptopodin were studied. Results The foot process width (420-2270 nm) in patients with Alport syndrome was positively correlated with proteinuria significantly (r=0.765, P<0.01). Foot process width was significantly lower in patients with mild proteinuria [475(420-900) nm) compared with that in patients with heavy proteinuria [1520(480-2270) nm] (P<0.05). In Alport syndrome children with heavy proteinuria, the distribution of nephrin and podocin changed dramatically. In two children with shorter proteinuria period (1 year), dramatic distribution change of nephrin and podocin occurred, however, the foot process along with synaptopodin preserved. Conclusions Podocyte foot process effacement and injury of slit diaphragm participate in the mechanism of proteinuria in Alport syndrome. The injury of slit diaphragm seems to be an early event in the development of foot process effacement in Alport syndrome, which may guarantee early treatment.

Objectives To investigate the relationship between albuminuria and all-cause mortality and cardiovascular mortality in middle-to-old-aged Chinese population. Methods A total of 2500 residents aged more than 40 years old were selected using random cluster sampling in Shougang community, Beijing, and 2315 of them took part in the survey finally. Morning urinary samples were collected. Urinary albumin and creatinine were measured. Albumin to creatinine ratio (ACR) was calculated and used as an index of albuminuria. The subjects were grouped according to ACR: normoalbuminuria (NO, ACR< 30 mg/g), microalbuminuria (MI, ACR 30-299 mg/g), and macroalbuminuria (MA, ACR≥ 300 mg/g). Albuminuria (AL) group consisted of MI group and MA group. Cardiovascular risk factors were also investigated. Then all-cause mortality and cardiovascular mortality were collected after 4 years. The Cox model was used to analyze the relationship between albuminuria and all-cause mortality after adjusting for confounders. Results The prevalence of microalbuminuria and macroalbuminuria was 7.6% and 1.4% respectively. After 4 years follow-up, the cardiovascular mortality was 2.7/1000 person-years in NO group, 19.9/1000 person-years in MI group, and 11.5/1000 person-years in MA group and the all-cause mortality was 6.6/1000, 25.9/1000 and 57.5/1000 person-years respectively. After adjusting for age, gender, smoking, body mass index, serum lipids, hypertension, diabetes mellitus, cardiovascular disease at baseline and serum creatinine, the hazard ratio (HR) of cardiovascular mortality in AL group was 5.26 [95% confidence intervals (CI) 2.26-12.24] compared with NO group; the HR of all-cause mortality was 3.34(95%CI 1.82-6.15). Among patients without cardiovascular disease at baseline, the corresponding HRs were 6.92 (95%CI 1.80-26.58) and 2.85(95%CI 1.22-6.65) respectively. Conclusion In the population studied, albuminuria is an independent risk factor for all-cause mortality and cardiovascular mortality.

Objective To investigate the clinical and renal pathologic features of isolated hematuria in children and the relationship between them. Methods A retrospective review of 251 cases of isolated hematuria undergone renal biopsy from 1995 to 2008 in our hospital were conducted to analyze their clinical manifestations and renal pathologic features. Results Among the pathologic changes, minor abnormalities was found in 93 cases (37.05%), normal biopsies in 62 cases (24.70%), IgA nephropathy(IgAN) in 52 cases(20.72%), thin basement membrane nephropathy (TBMN) in 17 cases(6.77%), mesangial proliferative glomerulonephritis(MsPGN) in 16 cases (6.37%), focal segmental glomerulosclerosis(FSGS) in 5 cases(1.99%), focal proliferative glomerulonephritis (FPGN) in 5 cases (1.99%), capillary proliferative glomerulonephritis (EnPGN) in 1 case (0.40%). IgAN was more popular in gross hematuria group than that in microscopic hematuria group (31.88% vs 16.48%, P<0.05). According to Haas classification, the ratio of class Ⅲ in two groups had no statistical significance (microscopic vs gross: 16.67% vs 4.55%, P>0.05). In the 35 cases (102 cases were detected) with elevated urinary microalbuminuria, the proportion of IgAN Ⅲ was significantly higher than those cases without urinary microalbuminuria (14.28% vs 0%, P<0.01). There were more FSGS and FPGN (total 20.00%) and less minor abnormalities (28.57%) in these cases as compared to the normal albuminuria cases (1.49% and 58.21%, all P<0.01). Conclusion The main pathologic changes of isolated hematuria in children are minor abnormalities, normal and IgAN. IgAN is more popular in the cases with gross hematuria. Elevated urinary microalbuminuria may be an indicator of more serious pathologic changes in children with isolated hematuria.

Objective To investigate the association between albuminuria incidence and blood pressure (BP) level or body weight index (BMI) in patients with essential hypertension from five regions in China. Methods A total of 5021 non-diabetic patients with clearly diagnosed essential hypertension were enrolled in our study. The participants came from five cities in China. Urinary albumin/creatinine ratio was measured in these patients for two times. The associations of albuminuria with BP level and BMI were analyzed. Results (1)There was no significant difference of albuminuria incidence between <60-year-old and ≥60-year-old patients. The longer the hypertension exited, the higher the albuminuria incidence was. (2) The albuminuria incidence was associated with blood pressure level significantly. The urinary protein excretion increased with the level of blood pressure. The albuminuria incidences in patients with normal BP, upper range of normal BP, Ⅰ, Ⅱ or Ⅲ stage hypertension were 26.3%, 27.3%, 28.7%, 31.5% and 40.3% respectively. (3) The albuminuria incidence was significantly different in patients with uncontrolled BP (≥140/90 mm Hg) compared with those with well controlled BP (<140/90 mm Hg) （27.1% vs 30.2%，P<0.05）. (4) The albuminuria incidence was higher in obese patients compared with those with normal BMI at same BP level, but the difference was not statistically significant. (5) Patients with albuminuria had more cardiocerebral or renal events as compared to those without proteinuria. 　 Conclusions The albuminuria incidence of non-diabetic hypertensive patients from 5 cities in China is 28.8%, of which the microalbuminuria incidence is 18.6% and the clinical albuminuria incidence is 10.2%. Uncontrolled BP is an important risk factor of proteinuria.

Objective To identify the accurate measurement of glomerular filtration rate (GFR) in chronic glomerulonephritis（CGN）patients. Methods Forty-two patients were enrolled in the study, including 15 females with age from 18 to 73 years old (mean 46 years old) and 27 males with age from 20 to 77 years old (mean 48 years old). The methods used for measuring GFR were classical dual plasma sample clearance method (tGFR), considered to be the gold standard, renal dynamic imaging method(dGFR), 24-hour creatinine clearance method (24hCcr). The difference and correlation amony them were analyzed. When the difference was significant, Pearson correlation and linear regression analysis were further performed. The difference of GFR detected by dGFR between left and right kidney of patients was compared simultaneously. A two-sided P value<0.05 was considered as statistically significant. Results Either dGFR or 24hCcr was statistically different from tGFR, but had excellent correlation with tGFR, and the coefficient was 0.916 (P=0.000) and 0.957(P=0.000) respectively. The linear regressions correlation existed and the regression equations were tGFR=0.936 dGFR-4.648 (F=208.941, P=0.000), tGFR =0.887 24hCcr+2.919(F=376.513, P=0.000) respectively. Difference had not statistically significance between left and right kidney of patients (P=0.591). Conclusions Neither dGFR nor 24hCcr can substitute tGFR, but both can reflect the GFR of the CGN patients safely and effectively. The decrease of GFR is homochronous for left and right kidney of CGN patients. Therefore, the 24hCcr can be chosen to evaluate the GFR in the hospitals without SPECT.

Objective To investigate the protective effect and mechanism of taurine on the cytotoxicity of iohexol on HK-2 cells. Methods HK-2 cells were exposed to iohexol at different dosage (25, 50, 100, 125 gI/L) for 6 h and at the dose of 100 gI/L for different time(2 h, 4 h, 6 h). Then taurine（3，12，24 mmol/L） was coincubated with iohexol (100 gI/L) for 6 h. Cell viability was assessed by CCK-8 assay. Cell apoptosis was determined by Hoechest 33342 flurescence stains，flow cytometry with AnnexinⅤ-FITC/PI double stains and caspase-3 activity by colorimetric assay. Bcl-2 and Bax expression were examined by Western blot. Intracellular ROS was detected by flow cytometry with fluorescent probe DCFH-DA. Results Iohexol decreased HK-2 cell viability and induced apoptosis in concentration-dependant and time-dependant manner (all P＜0.05). ROS was increased following iohexol (100 gI/L for 6 h) treatment (P＜0.05). Taurine increased cell viability and attenuated apoptosis in dose-dependant manner. The cell viability levels in taurine intervention (3，12，24 mmol/L) group were significantly increased compared with that in iohexol treated group respectively [(88.00±1.00)%, (91.33±0.58)%, (95.67±1.52) % vs (76.67±1.53)%, all P＜0.05]. Apoptosis rate by flow cytometry were decreased respectively [(8.84±1.75)%, (7.86±1.82)%, (6.30±1.50)% vs (11.98±0.39)%, all P＜0.05]. Caspase-3 activities were decreased respectively [(1.33±0.10), (1.27±0.06), (1.10±0.04) vs (1.42±0.13), all P＜0.05]. Taurine up-regulated the expression of Bcl-2, and decreased the intracellular ROS（all P＜0.05). Conclusions Iohexol induces cell apoptosis and oxidative stress. Taurine attenuates direct cytotoxic effect induced by iohexol. The anti-oxidative stress effect and up-regulated Bcl-2 expression may partly account for the protection of taurine.

Objective To observe the change of intima/media thickness ratio and expression of inflammatory factors in renal small artery of diabetic rats, and to explore the correlations of intim/media ratio with inflammatory factors and vascular lesions of diabetic nephropathy(DN) rats. Methods Seventy healthy SD rats were randomly divided into diabetic nephropathy group(DN, n=40) and normal control group(N, n=30). DN rat model was induced by intraperitoneal injection of streptozotocin (STZ). Thirty-five DN rats were successfully established. N group received same dose of citrate buffer. Rats were sacrificed after 4, 12, 24 weeks respectively. The intima/media thickness ratio in renal small artery was detected by immunofluorescence. The monocyte chemoattractant protein-1 (MCP-1) protein and mRNA expression of renal small artery were detected by immunohistochemistry and in-situ hybridization at each time point. Results Blood glucose and urine protein excretion (24 h) at different time points in DN group were significantly higher than those of N group (P<0.05). From the 12th week, Scr, BUN, serum phosphorus were significantly higher than those of N group (P<0.05). At the 4th week, renal small artery had the expression of MCP-1 protein and mRNA. The expression increased gradually with time, reached the highest at the 24th week, and was significantly higher than that of N group at each time point (P<0.05). Immunofluorescence results showed that as compared to N group, in the first 4 weeks, intima/media thickness ratio in DN group was not different, at the 12th week the ratio was higher but without significant difference, at the 24th week the ratio was significantly higher (P<0.05). Small artery intima/media thickness ratio of DN group was positively correlated with MCP-1, cholesterol, triglyceride, serum phosphate (r=0.742, P<0.01; r=0.740, P<0.01; r=0.829, P<0.01; r=0.580, P<0.01). Conclusions The arterioles intima/media thickness ratio of early DN is significantly correlated with MCP-1, lipids and phosphorus. MCP-1 may be involved in the DN vascular disease.

Objective To observe the release of inflammation-related factors after angiotensin Ⅱ(AngⅡ) stimulation in rat tubular epithelial cells (NRK-52E), to analyze whether these effects were mediated by TLR4-MyD88 pathway, and to reveal the novel mechanism of injury by AngⅡ on NRK-52E cells. Methods After synchronization, cells incubated with AngⅡ(10-7 mmol/L) were used as the stimulation group, cells without stimulation were as normal control. To determine the role of TLR4 and the adaptor MyD88, equal number of NRK-52E cells was added with 10-5 mmol/L candesartan or 20 mg/L TLR4 blocking peptide for 1 h and then incubated with AngⅡ (10-7 mmol/L) respectively. RT-PCR was used to analyze TLR4 mRNA and MyD88 mRNA expression. Immunofluorescence and confocal microscopy were used to observe TLR4 protein expression. ELISA was used to detect the concentration of tumor necrosis factor-alpha (TNF-α) and heat shock protein 47(HSP47) in cell supernatant respectively. Results TLR4 and MyD88 were highly expressed in AngⅡ-induced NRK-52E cells (P<0.01), and the TNF-α and HSP47 levels were also increased markedly compared with control group (P<0.01). In NRK-52E cells that were pre-incubated with candesartan, TLR4 and MyD88 expression were obviously inhibited, subsequently, HSP47 and TNF-α production decreased remarkably compared with AngⅡgroup(P<0.01). TLR4 blocking peptide had the similar effect in a dose-dependent manner, in which its effect was dependent on inhibiting TLR4-MyD88 expression. Conclusion The mechanism of AngⅡ-induced injury effect on NRK-52E cells is related to the increase of TLR4-MyD88 activity, which is followed by the enhance of TNF-α and HSP47 expression. This process is inhibited by candesartan via modulation of innate immune pathway.

Objective To investigate the role of mitochondrial respiratory chain in the hyperpermeability of human peritoneal mesothelial cells (HPMCs) induced by high glucose peritoneal dialysis solution (PDS). Methods HPMCs was cultured in a 1:1 mix of DMEM and PDS containing 1.5%, 4.25% glucose for 24 hours. A 1:1 mixture of 160 mg/L glutathione and 4.25% glucose PDS was also added. Transmesothelial electrical resistance (TER) measurement was examined for detection of permeability damage in HPMCs. Immunostaining and Western blotting analysis were used to detect claudin-1 expression. Mitochondrial superoxide (MitoSOX) Red staining and respiratory chain complexes activities were determined for detection of mitochondrial reactive oxygen species (ROS) production and mitochondrial complexes activities. Results TER was decreased in a time- and concentration-dependent manner after culture with high glucose PDS for 24 hours[4.25% glucose PDS group: from (34.7±1.5) Ω&#8226;cm2 to (4.3 ±1.4) Ω&#8226;cm2]. Claudin-1 was also down-regulated significantly by high glucose PDS (P<0.01). Complex Ⅲ activity was inhibited (10.8% of control, P<0.01) accompanied with increased mitochondrial ROS generation. These changes were partially prevented by glutathione. Conclusion Mitochondrial respiratory complex Ⅲ pathway has crucial importance in maintaining TER of HPMCs, which may reveal a valuable target for novel therapies to fight hyperpermeability of peritoneum during the prolonged PD treatment.

Objective To study the effect of recombinant human edostatin on peritoneal angiogenesis in uremic peritoneal dialysis(PD) rats. Methods Forty male SD rats were randomly divided into 5 groups: normal control rats (group 1), renal failure without PD rats (group 2), rats dialyzed with 4.25% PD solution (group 3), rats dialyzed with 4.25% PD solution and received subcutaneous injection of recombinant human endostatin 10 mg/kg (group 4), rats dialyzed with 4.25% PD solution and received subcutaneous injection of recombinant human endostatin 40 mg/kg (group 5). Recombinant human endostatin was given every other day during peritoneal dialysis period, total 14 times. After regular PD for 28 days, tissue immunohistochemical staining and RT-PCR were used to detect the mRNA and protein expressions of VEGF and bFGF in peritoneal tissues of each group rats. Microvessel density (MVD) of peritoneum was detected and quantified with anti-CD34 immunohistochemical staining. Results The mRNA and protein of VEGF and bFGF were expressed in each group. Compared to group 1, the mRNA and protein expression of VEGF and bFGF were significantly up-regulated in group 2 and group 3(all P<0.05). Compared with group 3, the mRNA and protein expression of VEGF and bFGF were significantly down-regulated in group 4 and group 5 (all P<0.05). Compared with group 4, the mRNA and protein expression of VEGF and bFGF were significantly down-regulated in group 5(all P<0.05). The new microvascular vessels in group 1 showed little or none. Compared with group 1, MVD was significantly increased in group 2 and group 3 (P<0.05). Compared with group 3, MVD was significantly decreased in group 4 and group 5 (all P<0.05). Conclusions Recombinant human endostatin can effectively inhibit rat peritoneal neoangiogensis. Down-regulated expression of VEGF and bFGF in peritoneum may be one of the mechanisms of recombinant human endostatin inhibiting peritoneal angiogenesis.