Objective To detect the proteins structure encoded by COL4A4 gene with different missense mutations of thin basement membrane nephropathy(TBMN) and to analyze the effect of gene mutation on the secondary structure of α4(Ⅳ) chain and its association with phenotype． Methods A COL4A4-1inked TBMN patient with FSGS by a missense mutation (g. 1214G>A resulting in p. G405E) diagnosed by clinical manifestations, family history and renal biopsy examination, as well as two controls (one healthy, one pure TBMN carrying a g. 1550G>A mutation resulting in p. G448S) were enrolled in this study. The fragments of cDNA with the two mutations and that of corresponding cDNA from the healthy control were expressed in E. coli. The secondary structures of recombinant polypeptides were analyzed by circular dichroism (CD) spectroscopy. Results CD spectra of healthy control exhibited a negative peak near 208 nm whereas that of TBMN patient with FSGS exhibited a negative peak near 220 nm. Furthermore, the magnitude of the negative peak of this patient decreased as compared with that of healthy control. CD spectra of pure TBMN control was slightly changed with the negative peak remaining near 208 nm and the magnitude slightly decreased as compared with that of healthy control. In addition, the secondary structure of polypeptide from healthy control was composed of about 1/4 α-helix and 1/4 β-sheet, whereas that form the patient presented about 1/3 α-helix without any β-sheet. The secondary structure of polypeptide from pure TBMN control was almost the same as the healthy control, except a slight reduction of α-helix and a slight increase of β-sheet. Conclusions Although the glycine substitutions exists in the nearby domain of α4(Ⅳ)chain, the TBMN patient complicating FSGS with severe phenotype and g. 1214G>A mutation and the pure TBMN control with the mild phenotype and g. 1550G>A mutation are revealed with different secondary structures of α4(IV)chain. Moreover, the secondary structure change of α4(IV) chain is consistent with their corresponding phenotype severity.

Objective To investigate the prevalence of chronic kidney disease (CKD) and associated factors in Changsha county of Hunan province. Methods Using a stratified, multistage sampling, 1950 residents (older than 20 years old) from 3 towns of Changsha county were randomly selected to be interviewed and tested for the kidney damage indicators and the associated factors with CKD. Results Eligible data of 1727 subjects were enrolled in the study. After the adjustment of age and gender component, the prevalence of albuminuria was 8.5%, hematuria 5.1%, and reduced eGFR 1.5%. Approximately 14.6% subjects had at least one indicator of kidney damage, and the awareness rate was 16.5%. Age, hypercholesteremia, hypertriglyceridemia, hypertension and diabetes were independently correlated with albuminuria. Female, age, hypertriglyceridemia and hyperuricemia were independently correlated with reduced renal function. Female was independently correlated with hematuria. Conclusions The prevalence of chronic kidney disease is 14.6% and the awareness rate is 16.5% in suburban adult population of the central south area of China. The spectrum and correlated factors of CKD in this county undergoing fast economic development are close to those of Guangzhou and developed countries.

Objective To investigate the expression of inflammatory mediators in renal tubular epithelial cells in patients with diabetic nephropathy (DN) and to explore the possible clinicopathological significance. Methods Twenty-three patients with DN diagnosed by renal biopsy and 10 patients with renal cell carcinoma undergone nephrectomy were allocated into DN group and control group, respectively. The renal expression of NF-κB p50, NF-κB p65, NF-κB p65 mRNA, MCP-1, OPN, α-SMA, and FN was detected by immunohistochemical or in situ hybridization assay. Serum creatinine, urinary N-acetylglucosaminedase (NAG), urinary albumin and 24-hour urinary protein were detected. The correlation between these inflammatory markers and clinicopathological data were analyzed. Results (1)Among all the 23 DN patients, granular degeneration of the renal tubular epithelium, focal tubular atrophy, infiltration of inflammatory cells and interstitial fibrosis were apparent, and none of these were found in control group. (2)Immunohistochemical and in situ hybridization assay showed that, compared with control group, expression of these factors increased significantly in renal tubular cells or interstitium in DN patients, and expression of α-SMA or FN was not found in tubular epithelial cells. (3)Statistics assay showed the tubular NF-κB p65 protein expression was correlated with all of the following factors: NF-κB p50 protein(r=0.792) and NF-κB p65 mRNA (r=0.763), tubular MCP-1 (r=0.825) and OPN (r=0.869) expression, interstitial α-SMA (r=0.327) and FN (r=0.432) expression, proteinuria(r=0.710), estimated glomerular filtration rate (eGFR) (r=-0.728), and urinary NAG (r=0.930), P<0.01 respectively. Conclusion Tubular inflammation may play a role in the pathogenesis and progression of DN.

Objective To identify the outcome of pregnancy and the alteration of renal function in women with nephrotic syndrome. Methods From 2003 to 2007, 59 pregnant women with nephrotic syndrome in our hospital were enrolled in the study. Their clinical data were retrospectively analyzed, including the time of kidney disease onset, 24-hour proteinuria, serum albumin, serum creatinine, blood uric acid, blood pressure, fetal survival, fetal mortality, rate of premature delivery, birth weight of the newborn, and proteinuria, renal function, blood pressure of the patients during their postpartum follow-up. Logistic regression analysis was used to identify the risk factors influencing the outcome of the patients and the newborns. Results The average gestational week was (20.35±9.40) weeks when proteinuria was detected in these pregnant women. The 24-hour proteinuria ranged from 3.5 to 15 g/24 h (median 5.1 g/24 h). The serum albumin was between 10 and 28 g/L(median 22.5 g/L). The serum creatinine was between 32 and 825 μmol/L (median 84 μmol/L) and the serum uric acid ranged from 196 to 793 μmol/L (median 385.5 μmol/L). Pregnancy-induced hypertension syndrome occurred in 75% of the patients, among whom 55.5% suffered from preeclampsia. Forty-three(72.9%) newborns survived , among whom 76.7% (33/43) were premature births and 62.8% (27/43) were low birth weight infants. 50% of the pregnant women still had nephrotic syndrome after delivery. 75% of 24 patients with pre-existing chronic glomerulonephritis had increased proteinuria during pregnancy. Among the 38 patients with renal insufficiency, 36.8% had poorer renal function after delivery. 23.7% of the patients progressed into end stage renal failure after delivery, 80% of whom had serum creatinine≥265 μmol/L. 89% of the patients had persistent hypertension after childbirth. The Logistic regression analysis indicated hyperuricemia during pregnancy (P=0.018, OR=1.012) and the increase of serum creatinine (P=0.039, OR=1.005) were risk factors of renal failure in pregnant women after delivery. Hyperuricemia(P=0.012, OR=1.006）was the risk factor of fetal death. Conclusions Pregnancy with nephrotic syndrome leads to a low fetal survival. Hyperuricemia is the most important risk factor of the poor outcome of pregnant women and newborn.

Objective To elucidate the clinical features of nutcracker syndrome complicated with IgA nephropathy (IgAN) and to increase its level of diagnosis and treatment. Methods Clinical data of 14 cases of nutcracker syndrome complicated with IgA nephropathy (patient group) and 36 cases of nutcracker syndrome (control group) were analyzed retrospectively. Nutcracker syndrome was diagnosed by ultrasonography and magnetic resonance angiography (MRA) and IgAN by renal biopsy. Differences of clinical data and images in two groups were analyzed. Results Gender, age and blood pressure of two groups were not significantly different. Higher Scr level [(81.2±21.3) μmol/L vs (61.2±11.8) μmol/L, P<0.01], more severe proteinuria [(1.1±0.6) g/d vs (0.3±0.2) g/d, P<0.01] and hematuria (2.3±0.9 vs 1.5±1.3, P<0.05) in patient group were found. Differences of ultrasonography and MRA in two groups were not significant. Conclusion Renal biopsy should be considered in cases of nutcracker syndrome with persistence of proteinuria, hematuria or abnormal morphology of urinary red blood cell.

Objective To elucidate the mechanism of angiotensin (Ang)Ⅱ in regulation of renin synthesis and secretion in primary cultured juxtaglomerular granular cells (JGCs). Methods Mice JGCs were isolated and cultured as described before. Real-time PCR was used to demonstrate the expression of angiotensin converting enzyme ( CE) and angiotencin receptor (AT1，AT2) in JGCs. Ang Ⅱ was co-cultured with JGCs stimulated by PGE2 and isoproternol or not. Renin activity in supernatant was detected by radioimmunoassay and renin mRNA expression was examined by real-time PCR. Different concentrations of AngⅡ were co-cultured with JGCs with different time (1 h, 4 h and 24 h), cAMP in cell lysate and supernatant was measured by Cayman Cyclic AMP EIA Kit. The cytoplasmic calcium was decreased by BAPTA-AM, and increased by thapsigargin and cyclopiazonic acid, which could be used to observe the cAMP concentration affected by calcium. Adenylyl cyclase type 5, 6 (AC5, AC6) mRNA expression of JGCs co-cultured with AngⅡ was measured by real-time PCR. Results Real-time PCR confirmed the expression of ACE and AT mRNA in JGCs of WT mice. Angiotensin Ⅱ reduced renin secretion in primary cultures of JGCs [(370.6±36.9) vs (299.6±25.7) ng AngI&#8226;ml-1&#8226;h-1, P=0.014]. Angiotensin Ⅱ dose-dependently down-regulated forskolin-stimulated cAMP production in JGCs. Thapsigargin and cyclopiazonic acid decreased cAMP level. BAPTA-AM increased cAMP production obviously [(11.09±0.48) vs (3.55±0.47) nmol/L, P<0.01]. Ang Ⅱ inhibited AC5 mRNA expression by 42.12%, but did not inhibit AC6 mRNA expression. Conclusions Angiotensin Ⅱ can directly inhibit renin synthesis and secrestion maybe through reducing AC-stimulated cAMP levels. Crosstalk between calcium and cAMP system may exist in precise regulation of renin in JGCs.

Objective To observe the effect of Xuebijing injection on the expression of tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 protein (HMGB-1) in rat peritoneal mesothelial cells (PMCs) induced by lipopolysaccharide (LPS). Methods PMCs were isolated from rat colic omentum and the 3rd generation cells were used in the experiment. PMCs were incubated with LPS at different concentrations (1,10,100 mg/L); with LPS (10 mg/L) for 2, 6, 12, 18, 21, 24, 36 h; with Xuebijing injection at different concentrations (2,10,20 g/L) after incubation with LPS (10 mg/L) for 2 h. PMCs in the control group were incubated with medium. HMGB-1 mRNA was detected by RT-PCR. TNF-α and HMGB-1 protein in supernatants was detected by ELISA. Results Compared to the control group, the expression of HMGB-1 mRNA and protein was significantly increased in groups stimulated by LPS in a time- and dose-dependent manner (all P<0.05); the expression of TNF-α was increased in the groups stimulated by LPS in a dose-dependent manner(P<0.05). In the groups stimulated by LPS (10 mg/L), the expression of TNF-α appeared double hump within 36 hours. Compared to LPS (10 mg/L) group, Xuebijing injection significantly inhibited the expression of HMGB-1 and TNF-α (all P<0.05） in a dose-dependent manner. Conclusions HMGB-1 as a late mediator of inflammatory responses may play a role in the pathogenesis of peritoneal dialysis related peritonitis. Xuebijing injection can reduce peritoneal inflammatory impairment by inhibiting the up-regulation of TNF-α and HMGB-1 induced by LPS.

Objective To study the expression and regulation of aquaporins (AQP) in cystic epithelial cells of jck mice with polycystic kidney disease. Methods Localization and regulation of AQP1, AQP2, AQP3 and AQP4 protein were analyzed by using the immunofluorescence and Western blotting. Results Kidneys of jck homozygous mice were 4 folds larger than those of litter matched wild-type mice. There were multiple cysts and fibrosis in the renal tissue of jck mice. The epithelial cells in cysts were flat in shape. Blood urea level in jck mice was (42.6 ± 6.7) mmol/L, which was 5 folds higher than that in wild-type mice [(8.4±1.9) mmol/L] (P<0.01). Immunofluorescence analysis showed that AQP1 was expressed in the apical and basolateral membranes of epithelial cells in proximal tubules, as well as in the thin descending limb of Henle and endothelial cells of descending vasa recta. There was no AQP1 expression in epithelial cells of cysts. AQP2 was expressed in the apical membranes of collecting ducts and renal cysts. AQP3 and AQP4 were expressed in basolateral membranes of collecting duct and renal cystic epithelial cells of jck mice. Western blot analysis showed the same protein sizes of AQP1, AQP2, AQP3 and AQP4 in both jck and wild-type kidneys. However, AQP1 expression was down-regulated in jck kidneys(P<0.01). Conclusion The renal cystic epithelia expresses AQP2, AQP3 and AQP4, which indicates that epithelial cells in renal cysts are derived from renal collecting ducts in jck mice and aquaporins may play an important role in renal cyst development.

Objective To explore the role of NG2 proteoglycan in the pathogenesis of glomerulosclerosis. Methods Eukaryotic expression vectors carrying the small hairpin RNA (shRNA) for NG2 mRNA , named as Psilencer-NG2, was constructed. Then, rat mesangial cells (RMC) were transfected with Psilencer-NG2, Psilencer-NC (negative control), pcDNA/NG2 (NG2 over-expressive vector) and empty vector pcDNAⅠrespectively. The expression of endogenous NG2 in RMCs was examined by real-time PCR and Western blotting. Cell proliferation was analyzed by MTT assay and flow cytometry. The expression of laminin was detected by real-time PCR. Results Transfection of pcDNA/NG2 into HBZY-1 cells resulted in over-expression of NG2 mRNA and protein(P<0.05, P<0.05). Transfection of Psilencer-NG2 led to reduced expression of NG2 mRNA and protein(P<0.01, P<0.01). The expression of laminin β1 significantly increased due to overexpression of NG2 and decreased by treating with NG2 siRNA. According to MTT assay, overexpression of NG2 significantly stimulated the proliferation of mesangial cells while NG2 silencing inhibited it. NG2 increased the cell number in S phase and decreased the cell number in G0/G1 phase, while silencing NG2 induced the decrease of cell number in S phase and the increase of cell number in G0/G1 phase. Conclusion NG2 actively participates in the pathogenesis of glomerulosclerosis by stimulating proliferation of RMCs and increasing the deposition of ECM.

Objective To develop a model of type 2 diabetes with early renal injury on spontaneously hypertensive rats (SHR). Methods The 6-week old SHR were fed with the diets enriched with sucrose (20%, W/W), lard (10%, W/W), cholesterol (2.5%, W/W) and chleolate (1%, W/W) to induce insulin resistance. Hyperglycemia was developed by intraperitoneal injection of streptozotocin (STZ, 35 mg/kg). Wistar-Kyoto rats (WKY) were used as normal controls. Rats with plasma glucose (PGL) ≥16.7 mmol/L were diagnosed as diabetes. Eight weeks after the induction of diabetes, plasma triglyceride (TG), cholesterol (CHO), glucose, systolic pressure(SP), 24-h urine protein excretion (Upro) were examined in all the rats, and the homeostasis model assessment of insulin resistance (HOMA-IR) was analyzed. Renal pathological changes were studied by immunohistochemical staining and electron microscope. Results After 2 weeks on the high sucrose and fat diets, the model rats exhibited significant increase in basal PGL, TG and CHO levels as compared to control rats (P＜0.05, respectively). The insulin resistance was developed in model rats demonstrated by the higher HOMA-IR(5.03±0.38 vs 2.61±0.34, P＜0.05). At the end of the experiment, model rats were associated with hypertension. Upro level was significantly increased in model rats compared with that in controls [(57.58±16.54) mg/24 h vs (5.35±1.90) mg/24 h, P＜0.01]. The kidney hypertrophy index (KWI) was significantly increased in the model rats compared to controls (P＜0.05). Moreover, the diabetic model rats showed glomerular hypertrophy, foot process effacement, micro villous transformation, glomerular basement membrane (GBM) thickening. Conclusion A rat model is successfully established, which presents typical features of human type 2 diabetes and can be served as an ideal model to study the diabetic nephropathy.

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.