Objective To find out a more rational pathological classification criteria for renal injury in patients with type 2 diabetes mellitus. Methods The renal clinicopathological features of forty-nine type 2 diabetic patients with macroalbuminuria were collected and were compared retrospectively. The patients without diabetic renal disease were excluded. According to the pathological features, the patients were divided into two groups: typical diabetic glomerulopathy (DG) and atypical diabetes-related renal disease (ADRD). Results The renal biopsy revealed DG accounted for 59.2% of the patients, while the remaining 40.8% presented atypical renal injury defined as ADRD. In DG group, volume fraction of mesangium per glomerulus, glomerular basement membrane width, atrophic tubules index, intersititium injury index and prevalence of hyalinization of renal arteriole were higher; podocyte density per glomerulus was lower; duration of type 2 diabetes was longer; the level of fast blood glucose, systolic blood pressure, mean arterial pressure, proteinuria and prevalence of diabetic retinopathy (DR) were higher; glomerular filtration rate (GFR) was lower. In ADRD group, body mass index and prevalence of obesity were higher; dyslipidemia was more severe. GFR was negatively correlated with glomerular global sclerosis rate in both DG and ADRD group. Proteinuria was positively correlated with volume fraction of mesangium per glomerulus in DG. No correlation between proteinuria and pathological features was found in ADRD. DR (94.8%) and duration of type 2 diabetes over five years (90.7%) had high negative predictive value for DG. Conclusions Renal injuries in type 2 diabetes patients are heterogeneous. ADRD is an atypical renal injury in type 2 diabetes patients which is different from DG. DR and duration of diabetes are more helpful in predicting DG separating from ADRD.

Objective To compare the clinicopathological features between two kinds of obesity-related glomerulopathy (ORG). Methods Twenty-three patients with obesity-associated glomerulomegaly (OB-GM) and 22 patients with obesity-associated focal and segmental glomerulosclerosis (OB-FSGS) diagnosed by renal biopsy during 1998 to 2008 in our center were enrolled in this study. A retrospective analysis of clinical and pathological data was carried out. Results (1) All the patients in these two groups were with abdominal obesity. Most of them were middle-aged male. There were no significant differences in gender, age, body mass index and waist circumference between these two groups（P＞0.05）. The mean course of disease in OB-FSGS group was significantly longer than that in OB-GM group[（21.7±29.7）vs (6.8±9.3) months，P＜0.05]. (2) Metabolic syndrome was found in the most patients of these two groups, but there were no significant differences in the levels of serum glucose, triglycerides, HDL-cholesterol, uric acid and blood pressure between them（P＞0.05）. (3) The 24-hour urinary protein and Scr level in OB-FSGS group were significantly higher than those in OB-GM group[（2.49±1.58） vs (0.83±0.87) g/d, P＜0.05; (102.09±25.07) vs (87.84±20.63) μmol/L, P＜0.05]. The serum albumin level, creatinine clearance and urinary osmotic pressure in the former were significantly lower than those in the latter [(38.67±7.00) vs (44.05±3.55) g/L, P＜0.01; (95.78±37.83) vs (128.72±31.20) ml/min, P＜0.01; (678.72±91.76) vs (840.69±133.88) mmol/L, P＜0.01]. (4) The mean glomerular diameters of both OB-FSGS group and OB-GM group were increased, whose difference was not significant [(204.3±23.1) vs (205.3±14.3) μm, P＞0.05]. Conclusion There are significant differences in the mean course of disease, 24-h urinary protein excretion, serum albumin level and renal function between these two different kinds of ORG.

Objective To investigate the incidence, risk factors and outcome of acute kidney injury (AKI) following cardiac surgeries. Methods Clinical data of 1056 patients undergoing open heart surgery in Renji Hospital from January 2004 to June 2007 were retrospectively analyzed. Univariate and multivariate analyses were used to evaluate possible pre-, intra-, and post-operative parameters associated with AKI according to AKI Network (AKIN). Results Of the 1056 patients, 328 (31.06%) developed AKI. In-hospital mortality was 4.07% in all discharges while 11.59% in AKI patients (P<0.01). Multivariate logistic regression analysis revealed that increased age (OR=1.40), pre-operative hyperuricemia (OR=1.97), pre-operative left ventricular insufficiency (OR=2.53), combined surgery (OR=2.79), prolonged operation time (OR=1.43), post-operative circulation volume insufficiency (OR=11.08) were risk factors of AKI. Conclusions AKI is a common complication and associated with increased mortality following cardiac surgery. Increased age, pre-operative hyperuricemia, pre-operative left ventricular insufficiency, combined surgery, prolonged operation time, post-operative circulation volume insufficiency are useful in stratifying risk factors for the development of AKI.

Objectve To investigate the association between insertion/deletion(I/D) polymorphism of angiotensin converting enzyme (ACE) gene and autosomal dominant polycystic kidney disease (ADPKD). Methods Polymorphism of ACE gene was analyzed by polymease chain reavtion (PCR) in 103 ADPKD patients and 16 ADPKD family constellations including 35 patients and 30 non-ill people. Clinical data were collected and age of onset, hepatocyst, hypertension, urinary tract infecton, urinary concretion, hematuria were used as the main parameters to analyze the association between ACE gene polymorphism and ADPKD. Results The age of onset in DD genotype was 7.2 years younger than that in DI genotype [（31.90±11.41）vs (39.10±10.08) years, P<0.05] and was 14.25 years younger than that in II gene type [(31.90±11.41）vs (46.15±14.74) years, P<0.05]. The age of onset in I/D genotype was 7.05 years younger than that in II genotype [(39.10±10.08) vs (46.15±14.74) years, P<0.05]. There were significance differences of main clinical symptoms (hypertension, hematuria and urinary tract infection) among three genotype groups. In 11 family constellations, ACE gene polymorphism presented genetic linkage, but without significant difference (P＞0.05); the genotype distribution was not significantly different between ADPKD and non-ill people (P＞0.05）, as well as between man and woman （P＞0.05）; the DD genotype frequency was significantly higher in ADPKD patients with chronic renal failure（P<0.05）. Conclusions The age of onset in DD gentype is the youngest among three groups. The incidence of hypertension and hematuria in DI genotype is the highest. The ACE gene polymorphism in ADPKD family constellation does not provide diagnosis information. The ACE gene I/D polymorphism may not contribute to ADPKD. The DD genotype of ACE may be a risk factor of renal failure in the ADPKD.

Objective To study the arterial stiffness in non-diabetic pre-dialysis chronic kidney disease (CKD) patients and to explore the associated factors. Methods Automatic pulse wave velocity (PWV) measuring system was used to examine carotid-femoral pulse wave velocity (CFPWV) as the parameters reflecting central elastic large arterial elasticity. Vascular calcification was quantitatively evaluated by plain radiographic film of abdomen, pelvis and hands. Blood pressure, biochemical parameters and intact parathyroid hormone (iPTH) were routinely detected. Stepwise multiple linear regression analysis was used to assess the associated factors of arterial stiffness. Results Ninety-six non-diabetic pre-dialysis CKD patients and 30 healthy people were enrolled in this trial. CFPWV in stage 3, 4 and 5 CKD patients was significantly higher than that in healthy controls [(11.63±2.39) m/s, (11.70±2.80) m/s, (12.88±2.49) m/s vs（9.70±1.66）m/s , all P<0.05]. Stepwise multiple regression analysis demonstrated that age, mean arterial pressure, vascular calcification and iPTH were independent impact factors of CFPWV. Conclusions Arterial stiffness of large artery increases in non-diabetic pre-dialysis CKD patients. Age, mean arterial pressure, vascular calcification and iPTH are independent impact factors of CFPWV.

Objective To investigate the changes of expression of peroxisome proliferator-activated receptor γ (PPARγ) and its coregulators and monocyte chemotactic factor (MCP-1) treated with interleukin-1β (IL-1β), and to analyze the mechanism of interaction of these factors. Methods Renal tubular cells(HK-2 cells) were cultured in vitro. Total cellular RNA was isolated for real-time quantitative polymerase chain reaction (real-time PCR), nuclear extracts were prepared for Western blot analysis and EMSA. The supernatant was collected for ELISA after the treatment of IL-1β at different concentrations and time points. Results Under stimulus of different concentrations of IL-1β (0～20 μg/L) for 24 hours, the mRNA expression of PPARγ, SRC-1, SRC-2 and PGC-1 decreased significantly (P<0.05), meanwhile NCoR increased obviously (P<0.05). In further time-dependent experiment, the mRNA levels of SRC-2 and PGC-1 decreased by 57％ and 48％, respectively, at 1 hour after treatment with 10 μg/L IL-1β (P<0.05). The expression of SRC-1 decreased by 43％only after 2 hours (P<0.05). The expression of NCoR was not obviously changed until stimulated by IL-1β for 8 hours (2.17 folds, P<0.05), then it decreased slowly. In the same time-dependent experiment, Western blot analysis showed that IL-1β(10 μg/L) significantly decreased the protein level of PPARγ at 4 hours (P<0.05). ELISA analysis revealed that the secretion of MCP-1 kept on rising and reached the peak (160.56±2.80) ng/L at 8 hours(P<0.01), then decreased to (50.82±1.25) ng/L at 24 hours(P<0.01). IL-1β could down-regulate the DNA binding activity of PPARγ, and the activity of NF-κB was up-regulated. Conclusions PPARγ and its coregulators are closely related to MCP-1 and NF-κB during inflammation response in kidney. The activation of NF-κB by IL-1β leads to the decrease of PPARγ and its coactivators expression levels, however the expression of MCP-1 and NCoR in renal tubular epithelial cells is up-regulated. PPARγ together with its coregulators participate in the inflammation response in kidney.

Objective To observe the expression and localization of DJ-1 in renal fibrosis, and to investigate the expressions of E-cadherin, vimentin and the level of β-catenin tyrosine phosphorylation in human tubular epithelial cells. Methods In vitro, the human tubular epithelial cells (HKC cell line) were cultured with 10 μg/L TGF-β1 for 72 h. The protein expressions of E-cadherin, vimentin and DJ-1 were measured by Western blot. RT-PCR was used to detect the expression of DJ-1 mRNA. The intracellular distribution of DJ-1 was observed by confocal microscope. In vivo, Masson stain was used to evaluate the level of renal fibrosis. The expression and disposition of DJ-1 in renal tissue were detected by immunohistochemistry. HKC cells were transfected with pEGFP-N1-DJ-1 via lipofectamine 2000. The efficiency of transfection was detected by fluorescence microscope. The expressions of DJ-1, E-cadherin, vimentin and β-catenin tyrosine phosphorylation level in the transfected cells were detected by Western blot. Results The expressions of DJ-1 protein and DJ-1 mRNA were up-regulated in renal tubular EMT cells. Most of DJ-1 protein localized in cytoplasm, and some was in nucleus. After stimulation by TGF-β1, the expressions of DJ-1 protein both in cytoplasm and nucleus was greatly increased, especially in nucleus. In vivo, renal tissue expressed DJ-1 in tubular epithelia, but little expression in glomeruli. In renal tissue from 5/6-nephrectomized rats, DJ-1 expression was greatly increased. In the DJ-1 transfectants, the expressions of DJ-1, vimentin and β-catenin tyrosine phosphorylation level were up-regulated, but E-cadherin expression was suppressed. Conclusion The increased expression of DJ-1 may promote renal fibrosis.

Objective To investigate the effects of lipoprotein lipase activator, NO-1886, on the mRNA and protein expression of glycogen synthase kinase-3β (GSK-3β) in the kidney of diet-induced diabetic minipigs. Methods Fifteen Guangxi Bama minipigs were randomized into three groups: C group (n=5, with the normal control diet), DM group (n=5, with the high-fat and high-sucrose diet), and NO-1886 group (n=5, with the high-fat and high-sucrose diet supplemented with 1.0% NO-1886). Plasma glucose, insulin, triglyceride (TG), oral glucose tolerant test, creatinine and blood urea nitrogen were measured monthly. Urinary samples in the morning were used for determination of microalbumin at month 0, 2, 4 and 5. The mRNA and protein expression of GSK-3β were measured by real time PCR, Western blot and immunohistochemistry in the kidneys obtained at the end of month 5. Results Compared with the C group, levels of plasma glucose, insulin, triglyceride and microalbuminuria were significantly increased in the DM group. The mRNA and protein expression of GSK-3β were increased in the kidneys of diabetic pigs (mRNA 0.0272±0.0052, protein 1.1600±0.0463, P<0.01) as compared with those of normal pigs (mRNA 0.0125±0.0045, protein 0.1385±0.0664). Compared with the DM group, the concentrations of plasma glucose, insulin, triglyceride and microalbuminuria obviously decreased in the NO-1886 group. The mRNA and protein expression of GSK-3β were decreased in the kidneys of the NO-1886 group (mRNA 0.0162±0.0019, protein 0.8429±0.0408, P<0.05) as compared with that of the DM group. Conclusion NO-1886 can improve disorders of glucose and TG metabolism and insulin resistance, and down-regulate the expression of GSK-3β in the kidneys, and protect renal function and morphologic damage in diet-induced diabetic minipigs.

Objective To verify whether the periodic or continuous exposure to high glucose may have different effects on human umbilical vein endothelial cell (HUVEC）apoptosis, and to explore the effect of NF-κB pathway on apoptosis of HUVEC induced by high glucose using the RNAi adenovirus vector. Methods RNAi combinant adenovirus vector which targeted 1566 site of NF-κB p65 mRNA was constructed and the effect of p65 gene knockdown in HUVEC was detected by Western blot analysis. Then, the RNAi adenovirus was transducted to explore the role of NF-κB pathway on the regulation of apoptosis in HUVEC induced by high glucose. The apoptosis of HUVEC was tested by flow cytometry and TUNEL assay. Results High glucose could induce apoptosis of HUVEC. p65 protein expression of nuclear extracts was significantly increased in high glucose culture as compared to control group, but only slightly increased in NF-κB-specific knockdown group, which maintained at basal state. Compared with normal glucose group, the number of TUNEL-positive cells in high glucose group was significantly increased (25.81％±1.77％ vs 8.20％±0.63％, P<0.05). The number of TUNEL-positive cells was decreased in 30.5 mmol/L glucose plus Ad-1566 than that in 30.5 mmol/L glucose plus Ad-DEST (11.49％±0.92％ vs 26.10％±0.98％, P<0.01). Flow cytometry and TUNEL assay showed that the apoptosis of human umbilical vein endothelial cells induced by high glucose was inhibited by the RNAi adenovirus. Conclusion High glucose induces apoptosis of HUVEC. Knockdown of NF-κB p65 may protect HUVEC from apoptosis by preventing high glucose-induced NF-κB nuclear translocation.

Objective To observe the Klotho expression in kidneys and renal tubular epithelial cells apoptosis in spontaneously hypertensive rats（SHRs） and the effects of cordyceps sinensis(CS), in order to study the mechanism of protective effects of CS on renal tubular cells apoptosis in hypertensive renal damage. Methods Twenty 22-week-old male SHRs were randomly divided into four groups: model (SHR) group, CS group (treated with CS, 5 g&#8226;kg-1&#8226;d-1), Los group (treated with losartan, 50 mg&#8226;kg-1&#8226;d-1) and CS+Los group (treated with CS 5 g&#8226;kg-1&#8226;d-1 and losartan 50 mg&#8226;kg-1&#8226;d-1). Five 22-week-old male Wistar-Kyoto rats (WKY) were used as the control group. After 8 weeks, the levels of 24 hours urinary protein (Upro), urinary N-acetyl-β-D- glucosaminidase (NAG), serum creatinine (Scr), blood urea nitrogen (BUN) and renal pathological changes were detected; the mRNA expression of Klotho, p53 and p21 was detected by RT-PCR; the protein expression of Klotho, p53, p21 and cleaved-caspase-3 was tested by Western blotting. TUNEL assay was applied to evaluate the renal tubular cell apoptosis. Results As compared to SHR group, the levels of 24 h urinary protein content [(52.16±29.3) mg, (49.97±32.5) mg, (54.67±30.09) mg vs (96.52±36.94) mg], urinary NAG[(44.13±9.11), (42.75±8.33), (41.96±7.88) U/L vs (54.07±6.57) U/L], Scr [(45.25±9.55）, (43.76±8.65), (45.18±7.28) μmol/L vs (53.84±10.21) μmol/L]and BUN [(8.25±1.03), (8.40±1.58), (8.32±0.98) mmol/L vs (8.91±1.24) mmol/L]were decreased(all P＜0.05), renal pathological changes were relieved, the levels of Klotho expression were up-regulated and the levels of p53 and p21 expression and cleaved-caspase-3 protein expression were down-regulated (all P＜0.01), tubular cell apoptosis was decreased [7.56%±0.52％, 7.93%±0.37％, 7.37%±0.36％ vs 13.32%±0.64％, P＜0.01] in CS, Los and CS+Los group. Conclusions Klotho, p53 and p21 play important roles in renal tubular cells apoptosis in hypertensive renal damage. CS can up-regulate Klotho expression, down-regulate p53 and p21 expression and decrease the cleaved-caspase-3 expression and tubular cell apoptosis to ameliorate the hypertensive renal damage.

Objective To investigate the protective effect of apocynin against renal oxidative injury in a rat model of hyperoxaluria. Methods Animal model of hyperoxaluria was established by administration of 0.8% ethylene glycol(EG) to adult male Sprague-Dawley rats in drinking water for 4 weeks. Simultaneous treatment with apocynin (0.2 g&#8226;kg-1&#8226;d-1) by intragastric administration were performed in the rats. Markers of oxidative stress(OS) state, urinary H2O2 and 8-(so-prostaglandin IP), and renal injury were assessed at the end of the study. Expression and localization of NADPH oxidase subunits (p47phox, gp91phox, Nox-1) in kidneys were examined by immunohistochemistry, real-time PCR and Western blot, respectively. Results p47phox expressed widely in kidneys of model rats, including renal cortex, inner medulla and outer medulla. Compared with the control, OS and renal injury occurred in rats receiving EG, in accordance with the up-regulated expression of NADPH oxidase subunits in kidneys. Treatment with apocynin significantly reduced the excretion of urinary H2O2 and 8-IP, improved the creatinine clearance and the kidney/body weight, with the down-regulated expression of NADPH oxidase subunits(except gp91phox mRNA) in kidneys, but the levels of OS markers in apocynin-treated rats were still higher than thoset of normal controls. Conclusions The increased expression of NADPH oxidase subunits is suggested to be partially accounted for the development of renal OS in this rat model of hyperoxaluria. Apocynin treatment is effective for renal protection in this model.