Objective To investigate the correlations between intima-media thickness(IMT) of carotid artery and its major risk factors in maintenance hemodialysis(MHD) patients. Methods A cross-sectional study was carried out in 75 MHD patients(MHD group) and 30 healthy volunteers(control group). IMT of carotid artery was examined by high-resolution B-mode ultrasonograph in all patients. The clinical and biochemical characteristics of MHD patients were collected. According to the IMT value of carotid artery the MHD patients were divided into three sub-groups: normal IMT group, abnormal IMT group and increased IMT group. Results IMT of MHD group was significantly greater than that of control group[(1.03±0.42) vs (0.63±0.11) mm, P < 0.01]. The differences of age, SBP, serum albumin, pre-albumin, cholesterol and serum phosphate between increased IMT group and normal IMT group were significant(P < 0.05 or P < 0.01). The SBP and serum phosphate level were also higher in abnormal IMT group than those in the normal IMT group(P < 0.01). Significant positive correlations were found between IMT and age(r=0.247, P=0.032), SBP(r=0.758, P<0.01), serum phosphate level(r=0.604, P<0.01). Significant negative correlations were found between IMT and serum albumnin(r=-0.292, P=0.011) as well as pre-albumin(r=-0.681, P < 0.01). The results of partial correlation test controlling for age were the same as the results of previous test. In multiple regression analysis, greater SBP(β=0.446, P < 0.01), lower serum pre-albumin level(β=-0.336, P < 0.01) and higher serum phosphate level(β=0.248, P=0.01) were significant independent risk factors for increased IMT, independent of other confounding risk factors. Conclusions IMT of carotid artery increased in MHD patients. Greater SBP, lower serum pre-albumin level and higher serum phosphate level are the significant independent risk factors for increased carotid artery IMT which may associate with advanced arteriosclerosis in MHD patients.

Objective To determine the prevalence of atherosclerotic renal artery stenosis(ARAS) in patients diagnosed and suspected coronary atherosclerosis by angiography and to find the correlative factors of ARAS through the analysis of their clinic data. Methods Coronary and renal corarteries of 126 patients were examined by angiography and clinical data were collected. Logistic regression was used to evaluate the association between renal artery stenosis(RAS) and clinical factors. Results Twenty-four (19.04%) of 126 patients examined by angiography had RAS, in whom 13(10.32%) with significant RAS. Twenty-four patients showed 28 renal artery stenosis(20 cases with unilateral, 4 with bilateral stenosis). RAS was found in the ostial(60.7%), in the main stem(35.7%) and in the branch(3.57%) of renal artery, respectively. Eighteen of 64 patients with and 6 of 62 without coronary atherosclerosis showed RAS. The result of logistic regression revealed that there was significant association between RAS and clinical factors such as coronary artery disease, hyperlipidemia(P<0.01), smoking and renal insufficiency(P<0.05). Conclusions The prevalence of ARAS is 19.04% in 126 patients with or without coronary atherosclerosis, but the prevalence of ARAS in the patients with coronary atherosclerosis is distinctly higher than that in the ones without. There is significant association between ARAS and coronary artery disease, hyperlipidemia, as well as smoking.

Objective To analyze the clinical and pathological characteristics of patients with MPO-ANCA positive Wegener granulomatosis(WG)and to investigate the difference between patients with MPO-ANCA and PR3-ANCA. Methods Patients with WG were selected according to both Chapel Hill Consensus Conference definition and American College of Rheumatology (ACR) classification criteria in 700 Chinese patients with ANCA-positive systemic vasculitis. The clinical manifestions were compared between patients with MPO-ANCA and with PR3-ANCA. Results Eighty-nine patients fulfilled the diagnostic criteria of WG, 54/89(60.7%) were MPO-ANCA positive, 34/89(38.2%) were PR3-ANCA positive, and the rest one was both positive. Patients with MPO-ANCA were female predominant compared with patients with PR3-ANCA, their male/female ratios were 23:31 vs 24:10, respectively(P < 0.05). Patients with MPO-ANCA also had multi-system involvement. However, the prevalences of arthagia, skin rash, ophthalmic and ear involvement were significantly lower in patients with MPO-ANCA than those in patients with PR3-ANCA (46.3% vs 70.6%, P <0.05; 20.4% vs 44.1%, P < 0.05; 27.8% vs 58.8%, P < 0.01; 40.7% vs 67.6%, P < 0.05; respectively). The prevalence of elevated initial serum creatinine was significantly higher in patients with MPO-ANCA than that in patients with PR3-ANCA (81.5% vs 61.8%, χ²=4.2, P < 0.05). The mean score of Birmingham vasculitis activity score (BVAS) was significantly lower in patients with MPO-ANCA than that in patients with PR3-ANCA(22.2±6.21 vs 24.7±6.87, P < 0.05). Conclusions Patients with MPO-ANCA positive WG were common in Chinese. The significance of patients with WG with different ANCA serotypes still needs further investigation.

Objective To identify an adequate measurement of glomerular filtration rate (GFR) in clinical practice. Methods ^99mTc-DTPA clearance (^99mTc-GFR) was measured as GFR marker in 101 patients with different chronic kidney disease. At same time GFR was determiued and calculated via the iohexol clearance (iohexol-GFR); the equation developed from the Modification of Diet in Renal Disease Study (MDRD-GFR)and Cockcroft-Gault formula(CG-Ccr)according to Scr. They were analyzed by correlation and regression, receiver operating characteristic (ROC) curve. Results According to the standard of NKF/K-DOQI, from the first to fifth stage of chronic kidney disease, the correlation coefficients for iohexol-GFR were 0.87, 0.89, 0.88, 0.86, 0.87 respecfiuerg(P < 0.001). The mean area under ROC curve of iohexol-GFR was 0.97; the correlation coefficients for MDRD-GFR were 0.80&#65380; 0.75&#65380; 0.71&#65380; 0.67&#65380; 0.56 respecfiuely(P < 0.01). The mean area under ROC curve of MDRD-GFR was 0.82; the correlation coefficients for CG-Ccr were 0.76&#65380;0.67&#65380; 0.62&#65380; 0.60&#65380; 0.53(P < 0.05). The mean area under ROC curve of CG-Ccr was 0.82. Conclusions Iohexol-GFR is the most accurate measurement among the three methods. The accuracy of MDRD-GFR is more accurate than CG-Ccr.

Objective To explore the molecular effect and interaction among nephrin, podocin, CD2AP and α-actinin-4. Methods Firstly, the recombinant RNA interference (RNAi) plasmid—psiRNA-hH1GFPzeo, specifically targeting to the mrna of nephrin, podocin, CD2AP or α-actinin-4, was respectively tansfected into the mouse podocyte clone (MPC5) to each knockdown (KD) the expression of nephrin, podocin, CD2AP or α-actinin-4. Molecular distributions were revealed by confocal microscopy, and the mRNA and protein expressions were detected with semi-quantitative RT-PCR and Western blotting. Results (1)In podocin KD group (siPod966 and siPod54), the mRNAs of podocin and nephrin were not detected, their protein decreased 92% and 79%, 82% and 67%, respectively. The mRNA and protein level of CD2AP increased 62% and 42%, 71% and 46%, respectively, whereas α-actinin-4 did not change. In nephrin KD group (siNep492), the mRNA expression and protein level of nephrin were not detected, CD2AP increased 35% and 48%, respectively; and whereas podocin and α-actinin-4 did not change. In CD2AP KD group (siCda744 and siCda21), the mRNA of expression CD2AP was not detected, and its protein level decreased 92% and 83%, the mRNA and protein of nephrin decreased 60% and 48%, 76% and 72%, respectively; podocin increased 38% and 22%, 56% and 44%, respectively; whereas α-actinin-4 did not change. In α-actinin-4 KD group (siAct1790 and siAct319), the mRNAs expression of α-actinin-4 and nephrin decreased 69% and 58%, 64% and 49%, respectively; their protein level decreased 81% and 55%, 71% and 64%, respectively. However, the mRNAs of podocin and CD2AP increased 50% and 34%, 45% and 28%, respectively; and their protein level increased 64% and 46%, 65% and 42%, respectively. (2) With their expression change, the distributions of nephrin, podocin and CD2AP shifted evidently from the cell membrane surface to the nucleus circumference, whereas α-actinin-4 showed no change, which was still localized in the cytoplasm and further extended to foot processes. Conclusion (1) Nephrin might more independently play a crucial role in the slit diaphragm complex. (2) Alpha-actinin-4 might interact directly or indirectly with nephrin, podocin and CD2AP. (3) The relationship among these podocyte molecules might not be spontaneous, either a single-directional or bi-directional reaction. (4) The normal localization of these podocyte molecules might depend on their normal expression quantity.

Objective To investigate the apoptosis of tubular cells after renal ischemia/reperfusion (I/R) injury in aging rats, and the effects of using butylated hydroxyanisole(BHA) to inhibit the products of ROS on the model. Methods Wistar rats aged 27 months were randomly divided into 4 groups, which were sham, I/R model, BHA gavage and nicardipine injection group. The model was established through clamping bilateral renal arteries 30 min for ischemia, and reperfusing them for 18 hours. The time points we chose were based on previous experiments. Renal function, pathological changes, apoptosis were observed, the expression of caspase-3 and Cyc C, the activity of caspase-3,MDA volume, activity of superoxide dismutase(SOD)in kidney tissue were examined. Results (1)renal function of all rats slipped following I/R injury, and the pathological changes were severe, and a lot of renal tubular epithelium turned into apoptosis. Expression of caspase-3 and Cyc C as well as MDA volume were increased, the activity of caspase-3 was increased and the activity of SOD was decreased.(2)Either BHA or nicardipine could ameliorate declines in renal function, pathological changes. Either of them could decrease MDA volume and increase the activity of SOD.Conclusions In aging rats, I/R injury leads to elevated ROS accumulation, which leads to progressive apoptosis. Both inhibition of L-type Ca²⁺ channel and activation of ROS can attenuate apoptosis in tubular cells.

Objective To investigate the effect of dehydroascorbate on reactive oxygen species in tubular epithelial cell induced by high glucose. Methods Tubular epithelial cell was cultured in RPMI-1640 medium containing 10% newborn calf serum. Uptake assays for AA and DHA in tubular epithelial cell was done by using the method provided by Robert Root-Bernsteinz. The intracellular formation of ROS was detected with the fluorescent probe CM-H2DCFDA by using confocal microscopy. Results At a DHA concentration of 1 mmol/L, increasing concentrations of glucose competitively inhibited DHA from entering into the cells such that the accumulation of DHA was smaller than half maximal at about 22 mmol/L glucose. Cytochalasin B, a kind of hexose transporter inhibitor, inhibited DHA from entering into the cells. At a glucose concentration of 25 mmol/L, DHA entry into cells was strengthened with increased DHA level, and DHA inhibited intracellular ROS formation in a dose-dependent manner when DHA level was less than 4 mmol/L. However, the inhibitory effect was not observed at 8 mmol/L of DHA. Conclusions Tubular epithelial cells are DHA dependent.VitC exclusion from tubular epithelial cells through competition of glucose and DHA for common transport mechanism will deprive the cells of the central antioxidant and this could lead to ROS accumulation.

Objective To observe the outcome of normal and senescent renal tubular epithelial cells after kidney ischemia/reperfusion injury (IRI), and investigate the role of cell senescence in the aging-related pathological changes in kidney. Methods Wild-type male mice at age of 2 (young group) and 12 months (aged group) were used to make ischemic models by clamping left renal hila for 45 minutes. At 0 d, 1 d, 3 d, 7 d, 1 month, 3 months and 6 month after reflow, renal tissues were examined for histomorphology, cell proliferation(PCNA), apoptosis and senescence (SA-β-gal). Results In both young and aged mice, renal tubule necrosis was the primary changes at day 0 after kidney IRI, but in aged mice, that was much more severe. Apoptotic tubular cells were detected at day 1 after IRI in young and aged mice, and they were much more widely distributed in aged mice. The most severe apoptosis occurred in tubular epithelial cells of both young and aged mice at day 7 after reflow (P < 0.05). In young mice, faint staining for SA-β-gal activity occasionally appeared in IRI kidney at month 1, and increased at month 3 and 6 after IRI(P < 0.05). No positive staining for SA-β-gal was noticed in contra-lateral kidney at any time mentioned above. Another pattern of SA-β-gal expression was detected in aged mice, both kidneys had widely positive staining for SA-β-gal at day 0 after IRI, but decreased notably at day 1 in the IRI kidney(P < 0.05), then increased again at month 3, but still less than that in the contra-lateral kidney, and more than that in the young mice at the same time point(P < 0.05). Six months after IRI, in both IRI kidney and contra-lateral kidney, positive staining for SA-β-gal almost reached the same level. Positive staining for nuclear PCNA in young and aged mice had no statistical significance (P > 0.05), although the number of positively stained nuclear PCNA were larger in young mice than that in aged mice. Correlation analysis between senescent and apoptotic cells in aged mice was made at day 1 after IRI, strikingly negative correlation was found between them (r=-0.82, P < 0.01). Conclusions IRI can promote the senescence process of normal tubular cells, and can accelerate the death(necrosis and/or apoptosis)process of senescent tubular cells. These variations may play an important role in the development and progression of aging-related pathological changes in kidney.

Objective To Compare the cost-effectiveness among hemodialysis(HD), CAPD and kidney transplantation(KT) in the first year and the second year.Methods Patients’data of above three groups in two years were retrospectively studied,including the cost,general conditions,hemoglobin level,influence on job and functional health status(by SF-36). Results The first year’s cost of KT group was higher than the other two groups, however, the second year’s cost was lower. There was no significant difference between HD group and CAPD group. The general conditions, the ratio of employment, the level of hemoglobin in KT group were better than those in HD group and CAPD group. Functional health status (SF-36): in MH dimension, VT and PF dimensions, the score of KT group and CAPD group was higher than that of HD group; in RP, GH, SF, RE dimensions, the score of KT group was higher than that of HD group and CAPD group(P<0.05), and the latter two groups were not significantly different.Conclusion KT is a less cost replacement therapy than HD and CAPD from the second year. The cost of HD and CAPD group is not significantly different within two years. The effectiveness of KT is better than HD and CAPD. The effectiveness of CAPD is a little better than HD. KT is the most cost-effective therapy as the increase of short-and long-term survival.

Objective To summarize the clinical, pathological features and inheritance mode of familial collagen type Ⅲ glomerulopathy. Methods The clinical manifestations and pathological findings of 2 affected brothers and their family information were collected. Results Two affected brothers, one was 33-year old and the other was 34-year old. Both of them had great amount of protein excretion in urine (3.1 g, 6.38 g respectively), and one of them had nephrotic syndrome. Both presented hypertension and renal insufficiency (serum creatinine 128μmol/L, 313 μmol/L respectively). Neither hematuria nor abnormalities of nail and bone was found. Their serum concentrations of procollagen Ⅲ peptide were elevated (>50 ng/L). Renal biopsy revealed that massive buddle fibrils were deposited in mesangium and glomerular basement membrane subendothelial area by electron microscopy. Strong staining of type Ⅲ collagen was observed in the mesangial area and along the glomerular capillary loops. Family survey showed their parents’^‘marriage was consanguineous. The concentration of procollagen Ⅲ peptide was also obviously elevated (>50 ng/L) in their younger sister but no proteinuria, hematuria, nor hypertension was detected and her renal function was normal. Conclusion Familial collagen type Ⅲ glomerulopathy is rare. Our findings supported an autosomal recessive pattern of inheritance. It is the first familial case reported in Chinese population.

Objective To screen for the COL4A3 and COL4A4 mutations in a Chinese consanguineous family with autosomal recessive Alport syndrome (ARAS). Methods Using PCR and direct sequencing, all 52 coding exons of the COL4A3 gene and 46 exons, except exon-1, of the COL4A4 gene were analyzed to detect mutations in the pedigree with ARAS. Furthermore, mutation was identified by restriction endonuclease AvaII in all other 20 members. Results A novel missense mutation (3725 G>A, G1242D ) in exon 42 of the COL4A3 gene was identified in homozygous form. This pathogenic mutation was demonstrated in heterozygous forms in all carriers in this family, whereas it was detected neither in the other normal members of the family nor in the 50 controls. In addition, 10 polymorphisms, including one nonglycine missense variants and 9 neutral polymorphisms, were detected in COL4A3/COL4A4. Conclusion The novel pathogenic mutation (3725 G>A, G1242D) of the COL4A3 gene may be the underlying pathogen in this family and it is the first reported case in ARAS.

Objective To investigate the mutations in Chinese families with Fabry disease. Methods Genomic DNA was extracted from peripheral blood cells of three probands diagnosed as Fabry disease and some family members. Seventy genomic DNA samples extracted from 70 unrelated normal persons were used as control. By PCR and direct sequencing, all 7 exons and their neighboring intronic sequences of the GLA gene of the probands were analyzed. Results Three mutations were identified in 3 probands: (1) deletion of 1 bp at nucleotide 1142 in exon 7 (1142DelG), leading to premature termination of protein translation at codon 390. (2) 902 G to A transition in exon 6 (codon 301), resulting in replacement of an arginine residue by glutamine (902G >A, R301Q). (3) 484 T to C transition in exon 3 (codon 142), resulting in replacement of a cysteine residue by arginine (484T >C, C142R). Mutation of GLA gene in 13 relatives of 3 probands was also screened and 6 cases with the same mutation as the relevant proband were found, including 5 heterozygotes and 1 hemizygote. Conclusion Three mutations including one novel mutation (1142DelG) are found in 3 Chinese families with Fabry disease by PCR-DNA sequencing.

Objective To investigate the effect of NPHS2 gene mutation of both V165X(467-468insT) and R168H (503G>A) on the expression and distribution of podocins in HEK293 cells. Methods The wild-type, V165X and R168H mutant cDNAs in the expression plasmids were transfected into HEK293 cells, and the subcellular localization of the wild-type, mutant podocins was studied by immunofluorescence staining with a specific podocin N-terminal antibody (P21) and a specific podocin C-terminal antibody (P35), immunolabeling and confocal microscopy. Results The fluorescence stainings of the wild-type, V165X and R168H mutant podocins with antibody P21 were positive. The fluorescence stainings of the wild-type and R168H mutant podocins with antibody P35 were also positive, whereas the staining of V165X mutant podocin with P35 was negative. The staining for wild-type podocin was distributed around nuclei and mainly on the cell membrane surface in a filamentous pattern, whereas V165X and R168H mutant podocins staining localized predominantly around nuclei with a loss of surface expression. Conclusions Both the molecular structure and the subcellular localization of V165X mutant podocin are changed evidently, so is the subcellular localization of R168H mutant podocin, whereas the molecular structure of R168H mutant podocin is little changed. The normal biological functions of the wild podocin rely on its normal molecular structure and subcellular localization as well.

Objective To investigate the effects of polycystin-1 N-terminal peptide (PC-1NTP) on proliferation, cell cycle and apoptosis of cystic-lining epithelial cells in human autosomal dominant polycystic kidney disease (ADPKD). Methods Cystic-lining epithelial cells were treated with PC-1NTP in vitro. MTT assay was used to detect the PC-1NTP effects on cells proliferation. Cell cycle and apoptosis were analyzed by flow cytometry. The mRNA expression of cyclinD1, p21^WAF1, bax, bcl-2 and MCM-2 were measured by fluorescence quantitative PCR. Results The growth and apoptosis of cystic-lining epithelial cells were significantly inhibited by PC-1NTP. The percentage of cells in G0/G1 phase increased, while decreased in S phase remarkably. The mRNA expression of p21^WAF1 and bcl-2 was significantly higher than that in the control group(P < 0.01), while the mRNA expression of cyclinD1,bax and MCM-2 was significantly lower than that in the control group(P < 0.01 or P < 0.05). Conclusions PC-1NTP resulted in G0/G1 phase arrest and elicited an attenuating effect on the proliferation and apoptosis of cystic-lining epithelial cells, which may be realized through regulating the expression of G1/S checkpoint regulation factor cyclinD1/p21^WAF1 and apoptosis regulating protein bcl-2/bax. PC-1NTP may be a new prospective maneuver in the treatment of ADPKD.

Objective To study renal involvement in hepatic glycogen storage disease(GSD) in childhood. Methods One hundred and eight patients aged less than 21 years old with typeⅠ a GSD (54 cases), typeⅢ (29 cases) and uncertain type hepatic GSD (25 cases). Urine analysis, urine albumin, urine protein of 24 h, urine β2-MG, BUN, creatinine, Ccr were evaluated. Results Of 108 patients with hepatic GSD, 16 patients (20.8%) had proteinuria proven by urine albumin or urine protein of 24 h, their ages first found proteinuria were 8~15 years. Two 15-year-old patients had proteinuria over 1.0 g/24 h. Among 72 patients, urine β2-MG of 51 cases (70.8%) increased (175~10 623 mg/L), and the mean urine β2-MG of typeⅠa GSD was much higher than that of typeⅢ GSD, 4138.2 and 1790.1 mg/L respectively. Of 91 patients, 10 had renal insufficiency, 1/10 (15-year-old girl) had heavy proteinuria (3.5 g/24 h), elevated BUN (9.3 mmol/L) and Scr (1061 μmol/L). Five elder patients (11~21 years old) had hematuria with renal colic caused by renal calculus. Conclusions Persistent protenuria, increased urine β2-MG, decreased Ccr, and renal stones are common complications of hepatic GSD in childhood. Renal function should be thoroughly evaluated during follow-up.

Objective To investigate the possibility of using serum and urinary endothelin-1 (ET-1), interleukin-6 (IL-6) assay as screening tools for atherosclerotic renal artery stenosis (ARAS). Methods Serum and urinary samples from 49 patients with ARAS, 32 cases with ≥2 risk factors for atherosclerosis and 30 normal controls were detected for ET-1 and IL-6 by RIA. The receiver operating characteristic (ROC) curves were then generated to assess their accuracy in screening ARAS by using selective renal angiography as golden standard. Results Urinary ET-1, urinary IL-6 and urinary-serum ratio of ET-1 in ARAS cases were higher than those in two control groups, and were all correlated with degree of renal artery stenosis. The area under the ROC curve of urinary ET-1, urinary IL-6 and urinary-serum ratio of ET-1 was 0.792, 0.756 and 0.779, respectively. The sensitivity and specificity of urinary ET-1 to distinguish ARAS (≥50%) was 80.0% and 72.8% respectively using 6.72 ng/mmol creatinine as the cut-off point. The cut-off value of ET-1 urinary-serum ratio was 12.59 with a sensitivity of 66.7% and a specificity of 61.7%. The sensitivity and specificity of urinary IL-6 were 73.3% and 70.4% respectively using 23.85 ng/mmol creatinine as the cut-off point. The sensitivity and specificity were improved to 80.0% and 77.8% respectively when using 12.60 ng/mmol creatinine as the cut-off point and combined with hypertension to perform series test. Conclusion Urinary ET-1 and urinary IL-6 could be used as screening tools for ARAS.

Objective To analyze the relationship between pathological diagnosis and clinical manifestation, and to investigate the importance of renal biopsy on the diagnosis of type 2 diabetic patients complicated with renal diseases. Methods The clinical and pathological data of 52 type 2 diabetic patients with abnormal urinalysis or increased serum creatinine who had renal biopsy performed in our department were studied. Results Among 52 patients, 32 cases were distinctly diagnosed as diabetic nephropathy (DN), including 3 presenting with diabetic nephropathy complicated with non-diabetic renal disease (NDRD), and 20 with the NDRD only. The preoperative diagnosis of 24 cases (46.15%) was accordant with the diagnosis after renal biopsy, while 10 cases (19.23%) were misdiagnosed. Between the two groups, except the differences in blood urea nitrogen, serum creatinine, diabetic duration and whether the patients were complicated with diabetic retinopathy, other clinical manifestations and laboratory findings were not significantly different. Conclusions Type 2 diabetes complicated with proteinuria may result from NDRD. It is difficult to differentiate the renal pathological changes merely by analyzing the clinical data. Therefore, renal biopsy is an important tool to confirm renal pathological changes.

Objective　To investigate the relationship between TGF-β1-induced fibronectin (FN) expression and up-regulation of integrin-link kinase (ILK) in human kidney tubular epithelial cells (HKC). Methods　Using cell culture and Western blot, the TGF-β1-induced expression of FN and ILK was tested. The ILK expression plasmid (pCMV-wtILK) containing wild-type full-length human ILK cDNA, or kinase dead ILK cDNA were transfered to HKC. The effect of overexpression of ILK on FN expression, and the blockage of ILK activation on the action of TGF-β1 on HKC were studied. Results　TGF-β1 induced FN expression of HKC in a dose-depended manner. In the time-course study, TGF-β1 upregulated ILK expression of HKC as early as 8 hours, meanwhile it induced FN expression. Overexpression of ILK in HKC tansfered with pCMV-wtILK could induce FN expression. Blockage of ILK activation abrogated TGF-β1-induced FN expression. Conclusions　A close relationship exists between TGF-β1-induced FN expression and upregulation of ILK in HKC. Blockage of ILK activation obliterates TGF-β1-induced FN expression.

Objective To investigate the role of caspase 3 in ATP depleted apoptosis in renal tubular epithelial cells(RTEC). Methods To induce apoptosis, REC were subjected to 60 min ATP depletion followed by recovery. Wild-type bcl-2 was overexpressed by infecting opossum kidney(OK) cells with adenovirus containing wild-type human bcl-2. Apoptotic cells were detected with Hoechst 33342 dye. Flow cytometry was used to quantify apoptosis and necrosis. Active caspase 3 and bcl-2 degradation were assessed by Western blot. Results ATP depletion and recovery resulted in activation of caspase 3 and the progressive accumulation of bcl-2 cleavage products and apoptosis. Overexpression of bcl-2 ameliorated apoptosis in the ATP-depleted RTEC followed by recovery. Caspase 3 reproduced the effect of caspase 3 on bcl-2 cleavage, whereas caspase 3 specific inhibitor DVED prevented bcl-2 cleavage in vitro. Conclusion Caspase 3 activition and mediated-bcl-2 degradation are likely to contribute to ATP depletion-induced apoptosis in RTEC.

Objective To investigate the signalling events follow the activation of RAGE in podocytes. Methods AGE and CML generated dichloroflurescin-sensitive intracellular ROS were measured by confocal microscopy. The activation of MAP kinases family were studied using Western blotting. MCP-1 mRNA expression was detected by semi-quantitative RT-PCR. Results Basal ROS was located in nucleus of starvation podocytes. AGE and CML rapidly generated intracellular ROS in podocytes. NAC pre-treated podocytes suppressed basal and inducible ROS generation and antibody for RAGE suppressed the inducible ROS. Blockage of ROS induced by NAC suppressed the expression of CML and H2O2-induced MCP-1. Phosphorylated extracellular signal-regulated kinase (ERK) was found in CML incubated podocytes at 10 min and was prevented by NAC or AFC. PD98058 Pre-treated podocytes partially inhibited the expression of CML-induced MCP-1 mRNA. No evidence were showed that p38 MAPK, SAPK/JNK, PI3K and PKC were involved in the signal transduction. Conclusion Activation of RAGE induces MCP-1 expression in podocytes via ROS-ERK signalling pathway.