Objective To study the effect of angiotensin-converting enzyme inhibitor(ACEI) on renal function in patients with unilateral atherosclerotic renal artery stenosis(ARAS). Methods From December 2002 to May 2004, 49 patients confirmed unilateral ARAS based on their clinical and angiography findings were divided into ACEI group(20 patients) and control group(29 patients). The baseline blood pressure was recorded and baseline Scr,BUN,Alb were measured to calculate eGFR using MDRD equation. Blood presure was measured monthly, and eGFR was assessed every 6 months during follow-up All patients also received renal duplex ultrasoundgraphy to measure intrarenal resistance index(RI) at baseline. Results There were no statistic differences of baseline characteristics between two groups. The average follow-up period was 9.9 months. When compared with baseline data, no changes occured in blood pressure for both groups (P > 0.05); eGFR remained stable in control group[(74.5±18.3 )ml·min^-1·(1.73 m² )^-1 vs (73.2±15.7)ml·min^-1·(1.73 m²)^-1, P > 0.05], whereas it is significantly decreased in ACEI group[(69.3±14.6)ml·min^-1·(1.73 m²)^-1 vs (63.5±16.4)ml·min^-1·(1.73 m²)^-1,P < 0.01]. At the end of observation, eGFR in ACEI group was lower [(63.5±16.4 )ml·min^-1·(1.73 m²)^-1 vs (73.2±15.7)ml·min^-1·(1.73 m²)^-1,P < 0.05], and the percentage of decrease of eGFR(△eGFR%) was higher(-9.1%±6.8% vs -0.6%±10.7%, P < 0.01) as compared with those of the control group. Pearson correlation analysis revealed that both baseline eGFR and contralateral RI were significantly correlated to △eGFR% (r=0.583 and -0.668, both P < 0.01). Conclusion ACEI could be harmful to renal function in unilateral atherosclerotic renovascular patients with obvious renal parenchymal injury,and renal function should be monitored during using ACEI.

Objective To analyse renal lesions associated with infectious endocarditis (IE). Methods Renal lesions associated with IE were reviewed. One hundred and fifty-five cases of IE were admitted to Peking Union Medical College Hospital from 1983 to 2004. C-square, t-test and Spearman’s rank correlation analysis were performed. Results One hundred and thirty-seven(84.4%) cases of renal lesions associated with IE with an average age of 38 were found. The ratio of male to female was 1.4 and the period of pre-renal lesion was 4.8 months. Renal lesions included asymptomatic hematuria and/or proteinuria (71.0%), acute nephritic syndrome(6.5%), nephrotic syndrome(2.6%), rapid progressive glomerulonephritis(1.3%), renal embolism(1.3%), isolated pyuria(3.2%), renal lesion not directly related to IE(2.6%). Acute renal insufficiency in 14 cases were caused by glomerulonephritis(5 cases), acute interstitial nephritis(5 cases), renal embolism(1 case), acute heart failure(5 cases) and the adverse effect of antibiotics(2 cases). Renal biopsy was taken in four patients. One diffuse proliferative glomerulonephritis, one membranous glomerulonephritis, one membrane-proliferative glomerulonephritis and one crescentic glomerulonephritis were found. All patients received antibiotic therapy and three of them stopped taking antibiotics, which was suspected to cause renal lesion. 20.4% cases received surgical therapy. 3.6% were treated with corticosteroid and/or immunoimpressive drugs and two cases of them were treated with intravenous bolus methylprednisolone. One case recieved anticoagulant therapy. 4.5% cases died. 43.8% cases with renal lesions were cured and 85.7% serum creatinine level decreased to normal. Statistical analysis showed that active treatment made no improvement on neither patients with or without renal lesion nor patients with different severity of renal lesion. Conclusions Renal lesions associated with IE are common. Most are asymptomatic hematuria and/or proteinuria. Acute nephritic syndrome, nephrotic syndrome, rapid progressive glomerulonephritis, renal embolism may also occur. It maybe appropriate to treat with corticosteroid, immunopressive drugs or intravenous bolus methylprednisolone for patients with rapid progressive glomerulonephritis under successful management of infective endocarditis.

Objective To investigate the influence of high glucose on connexin 43 expression and the gap junction intercellular communication (GJIC) function in mesangial cells. Methods The rat mesangial cells in vitro for 24 h or 48 h and were divided into normal glucose group(5.5 mmol/L), high glucose group(30 mmol/L) and osmotic control group (5.5 mmol/L glucose plus 24.5 mmol/L mannitol). Their GJIC function was measured by using fluorescence recovery after photobleaching assay(FRAP). The gene and protein expression of connexin 43 was detected by Northern blot, immunocytochemistry and Western blot. Results Normal glucose group expressed rich connexin 43 and kept a good intercellular communication,whereas intercellular communication decreased in high glucose group. The fluorescence recovery rate and recovery period of highglucose group were significantly decreased compared to those of normal glucose group. The mRNA and protein expression of connexin 43 was reduced in high glucose group as compared to those in normal group. Conclusion High glucose can inhibit the GJIC function by reducing connexin 43 expression in rat mesangial cells, which may be responsible for the dysfunction and abnormality of mesangial cells in diabetic nephropathy.

Objective To observe the process of renal tubular epithelial cell(TEC)transdifferentiation and the expression of Wilms tumor 1 gene(WT1) and to study the effect of WT1 on TEC transdifferentiation induced by IL-1α. Methods TECs were cultured in vitro and WT1 antibody was induced.Confluent cells were cultured for 5 days in DMEM/F12 media containing IL-1α(10 ng/ml), with or without 10 μg/ml of mouse anti-WT1 neutralizing antibody,and the ones cultured in DMEM/F12 medium only served as control. TEC morphologic and phenotype characteristics were observed by light microscope and electron microscope. The expression of α-smooth actin (α-SMA) and WT1 was detected by RT-PCR on day 1, day 2, and day 3 of cell culture. Results TEC cultured in media with IL-1α(10 ng/ml) for 5 days showed clear morphological and phenotypic changes. Many of them appeared fibroblast-like. Becoming elongated, the transformed cells showed marked hypertrophyand lost their cobblestone growth pattern. Scanning electron microscopy showed loss of apical-basal epithelial polarity and surface microvilli on the transformed cells. TEC cultured in media with (10 ng/ml) for 1 day started to re-express WT1 and α-SMA as well. However, WT1 became undetectable on day 3 in a transient pattern. α-SMA’s expression was enhanced in a time-dependent manner. TEC cultured in media with IL-1α(10 ng/ml) in the presence of 10 μg/ml of mouse neutralizing anti-WT1 antibody had little morphological changes, the WT1 and α-SMA’s expression clearly decreased in contrast to TEC cultured in media with IL-1α(10 ng/ml). Conclusions TEC embedded in high concentration of cytokine for long period would induce epithelial-myofibroblast transdifferentiation itself. The transient, re-expression of WT1 could play an important role in epithelial- myofibroblast transdifferentiation. The re-expression of WT1 in adult TEC could be the inner promotor of TEC transdifferentiation.Neutralization of WT1 gene product in vitro could counteract TEC transdifferentiation as well as kidney fibrosis.

Objective To investigate the effect of transforming growth factor-β1(TGF-β1) on the activity of connective tissue growth factor(CTGF) gene promoter in human renal proximal tubular epithelial cell line HK-2. Methods The regulation fragment of 5’ flanking region of human CTGF gene was linked to pGL3-Basic vector. The recombinant plasmid pCTGF-luc was transient transfected to HK-2 cells. The activity of CTGF promoter after treatment of TGF-β1 and mitogen-activated prontein kinases(MAPK) pathway inhibitors was assayed by means of luciferase reporter gene assay system. Results TGF-β1-induced increase of CTGF promoter activity was concentration-dependent, with a plateau at 5 ng/ml by 1.82-fold vs control(P < 0.05). The TGF-β1 stimulation of CTGF promoter activity was time-dependent, too. After exposure to TGF-β1(5 ng/ml), the maximal level of luciferase activity was reached at 12 h by 2.10-fold vs control(P < 0.05). Blockade of MAPK pathway with PD98059,the extracellular signal-regulated protein kinase (ERK) inhibitor, significantly increased basal activation of CTGF promoter in HK-2 and that of TGF-β1-induced in a range of concentration (0.5~10 μmol/L). SB203580, the p38MAP kinase (p38MAPK) inhibitor, markedly decreased TGF-β1-induced activation of CTGF promoter in a dose-dependent manner. However, inhibition of c-Jun-N-terminal kinase(JNK) of MAPK pathway by SP600125 was without effect. Conclusions TGF-β1 stimulates the transcriptional activity of CTGF gene promoter in HK-2 cells in a dose- and time-dependent manner. MAPK pathway(ERK and p38MAPK)may play a role in the regulation of TGF-β1-induced CTGF expression.

Objective To investigate the protective effect of hepatocyte growth factor (HGF) and its mechanism on renal interstitial fibrosis following unilateral ureteral obstruction(UUO) in rat kidney. Methods Rats were randomly assigned to UUO group, HGF treated group and sham-operation group(SOR). HGF was administered by peritoneal injection into rat with renal fibrosis disease induced by UUO. The levels of BMP7 and CTGF mRNA were examined by RT-PCR. The sites and the expression of BMP7 and CTGF were examined by immunohistochemistry staining and Western blotting. The protein expression of TGF-β1,α-SMA, and FN was detected by immunohisto-chemistry at each time point. Results The mRNA and the protein levels of BMP7 were significantly lower in UUO group than those in HGF treated group, whereas the mRNA and protein levels of CTGF were significantly increased in UUO group. Compared to UUO group, the levels of TGF-β1, α-SMA and FN were significantly decreased in tubulointerstitium of HGF treated group. Conclusion HGF counteractes renal interstitial fibrosis by blocking tubular epithelialmyofibroblast transdifferentiation through down-regulation of CTGF and up-regulation of BMP7 in vivo.

Objective To investigate the proinflammatory effects of glomerular fibrin deposition on inflammatory response and its mechanisms in rats. Methods Female Wistar rats were randomly divided into four groups(n=8 per group): normal control group (NC group) injected with saline alone; Lipopolysaccharide (LPS) group with fibrin deposition induced with LPS; TL group treated with LPS and tranexamic acid;UT group treated with LPS,tranexamic acid and urokinase. Tranexamic acid and urokinase were used to increase and decrease fibrin deposition, respectively. Fibrin and macrophages were detected by immunofluorescence and immunohistochemical staining. Monocyte chemoattractant protein-1(MCP-1) and vascular endothelial-cadherin (VE-Cad) protein and mRNA expression were analyzed by immunohistochemistry and RT-PCR, respectively. Results Fibrin deposition and macrophages of TL group were significantly higher than those of LPS, UT groups(P < 0.05). And there were significant differences between TL and LPS, UT in renal tissue MCP-1, VE-Cad mRNA and protein expression(P < 0.05). Conclusions Fibrin deposition contributes to renal inflammatory response. Proinflammatory effects of fibrin deposition may be involved in the control of MCP-1, VE-Cad expression.

Objective To characterize the expression of surfactant protein A(SP-A) in normal and acute pyelonephritic rat kidneys and to study the correlation of infection and inflammation with SP-A expression. Methods Twenty-one rats were randomly assigned into three groups:control, sham operation and pyelonephritic group. HE staining was used to determine tubulointerstitial inflammation. Reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blotting were used to determine the mRNA expression and protein level of SP-A. Immunohistochemical staining was used to label the localization and intensity of SP-A expression in kidney tissue. The correlation between intensity of SP-A expression and interstitial inflammation was also evaluated. Results In pyelonephritic group, tubulointerstitial inflammation was more prominent than that in control and sham groups (54.3±11.5,6.4±1.4, 8.6±1.9,respectively). RT-PCR and Western blotting revealed that SP-A expression was up-regulated in pyelonephritic group (in mRNA level: 2.2±0.58, 0.9±0.25, 1.1±0.30; in protein level: 0.45±0.09, 0.24±0.05, 0.26±0.05, respectively). Immunohistochemical staining demonstrated that SP-A expression was mainly localized on epithelial cells in outer medullary and collecting tubules in normal group and sham group, but strong staining extended to collecting tubules in pyelonephritic group. The tubulointerstitial inflammation score was positively correlated with the intensity of SP-A expression (r=0.67,P < 0.01). Conclusions SP-A expression can be observed in outer medullary and collecting tubules of the normal rat kidney. The SP-A level in the tissues of kidney is significantly higher in rat pyelonephritic model than that in either control or sham group. Moreover, significantly positive correlation exists between SP-A level and the degree of tubulointerstitial inflammation in this pyelonephritic model. SP-A may play an important role in host-defense and inflammation regulation in pyelonephritis.

Objective To investigate the effects of monocyte chemoattractant protein-1(MCP-1) on the cultured rat mesangial cells(MCs) proliferation and the expression of fibronectin(FN、collagen type Ⅰ(ColⅠ), and study intracellular mechanism through p38 mitogen-activated protein kinase (p38MAPK) signaling pathways. Methods After stimulation of different concentrations of MCP-1 and p38MAPK inhibitor SB203580, the cell proliferative activity was assessed by tetrazolium salt colorimetry assay(MTT). Semi-quantitive reverse transcription and polymerase chain reaction amplication(RT-PCR) was used to detect the mRNA expression of FN and ColⅠin mesangial cells.The protein contents of FN and ColⅠ in the supernatant were examined by ELISA. Results The cell proliferative activity, the mRNA and protein expression of FN and ColⅠwere increased with the higher concentration of MCP-1 and the longer incubated time. There were significant differences compared to the normal group in a dose- and time-manner(P < 0.01). When the p38MAPK inhibitor SB203580 was used, the above mentioned indexes were decreased significantly compared to each MCP-1 group(P < 0.01). Conclusions MCP-1 could promote the cell proliferation and up-regulate the expression of FN and ColⅠin rat mesangial cells cultured in vitro. The intracellular signaling transduction pathway is related to p38MAPK. MCP-1 plays certain role in the pathogenesis of mesangial cell proliferative glomerulonephritis(MsPGN).

Objective To investigate the effects and mechanisms of tissue inhibitor of metalloproteinase 1(TIMP-1) on the apoptosis of rat mesangial cells(MC) induced by high glucose.Methods Rat mesangial cells were cultured in medium 1640 with different glucose concentrations for 24 h, 48 h and 72 h respectively. The apoptosis ratio was tested by both flow cytometry AnnexinV/PI double stains and AO stains. MCs were transfected with empty pcDNA3.1 vector, TIMP-1S-pcDNA3.1(PCT), and TIMP-1AS-pcDNA3.1 (PCA) by liposomal transfection reagent. The expression of human TIMP-1, endogenous rat TIMP-1, bax and bcl-2 were examined by PCR and RT-PCR. Caspase-3 activity was analysized by CaspACE^TM Assay System. Results High glucose could induce apoptosis in a dose- and time-dependent manner. The coefficient of determination was r=0.925 and r=0.9867 respectively(P < 0.01). Exposed to high glucose (30 mmol/L), the apoptosis ratio of PCT group was 4.3%±1.11% at 24 h,and 7.78%±0.92% at 48 h, respectively. As compared to control(14.95%±1.6% for 24 h,43.03%±4.2% for 48 h) and empty vector groups, fewer MCs underwent apoptosis(P < 0.01) in PCT group. The apoptosis ratio of PCA group was 22.5%±1.6% at 24 h,and 53.68%±3.4% at 48 h, which suggested much more MCs in PCA group experienced apoptotic changes(P < 0.01). Under the influence of high glucose, PCT transfection could down-regulate the expression of bax, whereas PCA transfection could up-regulate the expression of bax by contrast. Compared with control group (the value of A was 0.1407±0.007), the activity of caspase 3 in PCT group decreased obviously (the value of A was 0.086±0.009, P < 0.01), while its activity increased greatly (the value of A was 0.186±0.02, P < 0.01) in PCA group. The exogenous human TIMP-1 seemed to have little effect on the expression of bcl-2 mRNA. Conclusions TIMP-1 can inhibit the apoptosis of MCs induced by high glucose,which partly involves in lowering the expression of bax and attenuating the activity of caspase-3.

Objective To investigate the renal protection of Chinese cobra venoms(CCV) and its mechanism in renal ischemia/reperfusion(I/R). Methods Thirty-two rats were divided into four groups. 0.1% CCV was separately infused into abdominal cavity at 0.5 h, 24 h before reperfusion in groupⅠandⅡ. Group Ⅲsuffered from kidney I/R was served as pathological control. Group Ⅳ was sham operation group. BUN and Scr were measured before ischemia and 24 h after reperfusion. Complement C3 was observed at 0, 0.5, 2, 24 h after reperfusion. The kidney samples were examined by HE stain under light microscopy. Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP-biotin in situ nick-end labeling(TUNEL). Results Significant histological damage, apoptosis of tubular cell and impaired renal function were found in groupⅠand Ⅲ.The above indexes decreased to a less extend in group Ⅱ(P < 0.05). Complement C3 in groupⅡ decreased markedly at the beginning of reperfusion and remained the same level since then, which is significantly different from that in group Ⅰ and Ⅲ. HE staining indicated severe lesion mainly located in the tubules in group Ⅰ and Ⅲ, whereas the severity was markedly relieved in group Ⅱ. A significant amount of TUNEL positive cells were observed in group I and Ⅲ as compared to group Ⅱ (P < 0.05). In group Ⅰ and Ⅲ, the C3 level significantly decreased at 2 h following reperfusion as compared to 0 h following reperfusion(P < 0.05). Conclusions I/R could lead to significant histological damage and renal dysfunction. Pretreating the rats with CCV 12 h before reperfusion could reduce C3 notably and prevent the kidney from c3 mediated I/R injury efficiently.

Objective To investigate the influence of pretransplant soluble CD30 (sCD30) level on acute rejection episode in kidney transplant recipients. Methods This retrospective study included 707 allograft kidney transplant recipients from Dec 1998 to Aug 2003 and 40 healthy as controls. The recipients′ pretransplant serum sample and post-transplant clinical data including acute rejection episodes were collected. The recipients were divided into low, median, high level sCD30 group according to their pretransplant sCD30 levels. sCD30 levels of the recipients and the controls were determined by ELISA assay in duplicate. Results Kidney graft recipients had a significant increased pretransplant serum sCD30 content as comprared to healthy controls(P < 0.01). The incidences of vascular rejection, cellular rejection and bordline increased positively with the pre-transplant sCD30 level (all P < 0.05). But the inversion rates of rejection were 100%, 90.6% and 78.6 in the low,median and high sCD30 groups,respectively. The sCD30 levels of vascular, cellular, bordline and clinical rejection groups were (198.95±76.09),(165.89±44.56),(172.94±74.22) and (161.23±64.87) U/ml,respectively, which were all higher than those of non-rejection group (all P < 0.05). The sCD30 values were the highest in the vascular rejection. Multifactors logistic regression analysis indicated that the sCD30, PRA and CMV antigen were risk factors of acute rejection with odds ratio of 2.683, 2.384 and 2.065, respectively. Conclusion Pre-transplant serum level of sCD30 is a sensitive prognostic and risk factor for acute rejections.

Objective To establish a new type of non-bacterial peritoneal fibrosis model in rats. Methods Thirty-four SD rats were divided into five groups: group1 (control), group 2 (normal saline), group 3 (4.25% dextrose solution), group 4 (4.25% dextrose solution with lipopolysaccharide), group 5 (4.25% dextrose solution with lactobionate erythromycin). Animals were sacrificed after 5 weeks. A 2-hour peritoneal equilibration test (PET) was performed. Dialysate-to-plasma urea ratio (D/Purea), glucose reabsorption (D₂/D₀), net ultrafiltration (UF) volume, the level of hydroxyproline and fibronectin were determined. Peritoneal membrane histology was evaluated by light microscopy. Results The D₂/D₀ ratio and net ultrafiltration volume in group 5 were significantly lower than groups 1 and 2 (P < 0.05). The D/Purea ratio in group 5 was significantly higher than groups 1, 2 and 3 (P < 0.05). The levels of hydroxyproline and fibronectin in group 5 were significantly higher than groups 1 and 2 (P < 0.05). The thickness of the peritoneal membrane in group 5 was significantly greater than that in groups 1, 2 and 3 (P < 0.05). The quantity of peritoneal vessels in group 5 was significantly greater than that in groups 1 and 2 (P < 0.05). Conclusion A new type of non-bacterial peritoneal fibrosis model in rats could be established by 4.25% dextrose solution with lactobionate erythromycin.