Objective To understand the situation of oxidative stress among diabetic nephropathy (DN) patients and observe the antioxidative effect of losartan at increasing dose in DN patients. Methods Thirty type 2 DN patients who neither smoked and nor took antioxidants were selected. The study began with an initial 4~6 weeks screening-treatment. Eligible patients then received losartan 50 mg/d daily for 8 weeks followed by losartan 100 mg/day daily for an additional 8 weeks. Blood glucose and blood pressure were closely monitored over the whole study period. All patients were followed up every other weeks，their 24-hour urine samples，fresh urine and venous blood sample were collected to measure urinary protein and creatinine excretion， urinary 8-OHdG，SOD，TAOC and MDA excretion，serum SOD，TAOC，MDA and other blood biochemistry parameters.Urinary 8-OHdG was determined by capillary electrophoresis and liquid phase chromatography. Results The total 24 hours urinary 8-OHdG excretion and the serum MDA concentration were higher than the normal values. The serum and urine SOD concentrations were lower than the normal values. There was an improvement in urinary 8-OHdG，serum and urine SOD， serum and urine MDA levels with losartan therapy. Compared with losartan 50 mg/d， the antioxidative effect of losartan 100 mg/d was more noticeable. Obvious decrease in 24-hour proteinuria on exposure to losartan was found， without severe adverse effect. Conclusions Oxidative stress damage is active in DN patients. Losartan has antioxidative effect on DN patients. Compared with losartan 50 mg/d， the antioxidative effect of losartan 100 mg/d is more marked，without increasing side effect. Losartan’s antioxidative effect may be involved in its beneficial mechanisms on DN.

Objective To observe the curative effect of total parathyroidectomy(PTX) with forearm autograft on the uremic patients suffering from severe secondary hyperparathyroidism. Methods To analyze the changes of intact parathyroid hormone(iPTH)， blood calcium and phosphate，blood haematocrit(Hct)，dyslipidemia， adequacy of hemodialysis(Kt/V、UUR)on 12 patients accepting PTX. Results The orientation of the parathyroid node detected by B-ultrasonic was consistent with location observed in surgery among 11 patients. The iPTH and blood calcium and phosphate decreased after PTX. And the operation also ameliorated the Hct， dyslipidemia and adequacy of hemodialysis. Recurrent hyperparathyroidism occurred in three patients among whom two patients received vitamin D therapy with the result of remaining iPTH in normal range and another one patient received resection of autograft resulting in normal iPTH. Conclusions PTX with forearm autograft is effective to treat severe secondary hyperparathyroidism in uremic patients. PTX can ameliorate anemia and dyslipidemia. The orientation of parathyroid node by B-ultrasonic before surgery is feasible. The “really” totally PTX and the proper tissue for autograft is crucial to avoid recurrence.

Objective To evaluate the association between the K469E polymorphism in the intercellular adhesion molecule-1(ICAM-1) gene and type 2 diabetic nephropathy (DN). Methods According to their urine albumin excretion rate(AER)，a total of 210 patients with type 2 diabetes were divided into type 2 diabetes with and without nephropathy groups. Another 98 controls with matched age and gender were selected from the same area. All patients and controls were genotyped by polymerase chain reaction (PCR) and digested with restriction endonuclease. Results The frequency of the KK genotype and K allele were significantly higher in the patients with diabetic nephropathy than those without nephropathy (genotype 36.4% vs. 22%，χ2=9.358 P=0.009； allele 60.9% vs. 46%，χ2=9.369 P=0.002). Compared with genotype EE and KE class， DN morbidity in genotype KK class was significantly increased. Logistic regression analysis showed that the KK genotype was an independent risk for diabetic nephropathy in our population. Conclusions K469E polymorphism of ICAM-1 is associated with type 2 DN， and K allele may be susceptible to DN.

Objective To explore the clinicopathological characteristics and prognostic risk factors of hypertensive nephrosclerosis. Methods A retrospective study was performed on 63 hypertensive patients with renal injury. The clinical date such as age， gender， family medical history， blood pressure，urine protein excretion，serum indicates，eyeground and echocardiography were analyzed. Patients were divided into 3 groups according to histological diagnosis:benign nephrosclerosis(BN)， malignant nephrosclerosis (MN) and primary nephritis (PN). The clinicopathological data were compared among three groups. The risk and prognostic factors were further analyzed for the patients confirmed by pathology. Results Among 63 patients，49 (77.8%) were diagnosed as HN， including 12 (19.0%) of MN， 37(58.7%) of BN， and 14 (22.2%) as PN. More males， older， frequent hypertension family history and longer duration were observed in BN group， whereas lower urine protein excretion was found in BN and MN groups as compared to PN group. MN group exhibited higher left ventricular mass index (LVMI)than PN group. 78% BN patients presented grade 0~Ⅱ hypertensive retinopathy and most of MN patients grade Ⅲ~Ⅳ. The frequency of globally sclerotic glomeruli increased significantly in PN group as compared to BN and MN groups， and so did the score of chronic index of renal pathology. The severity of vascular injury， assessed by the myointimal proliferation， was higher in BN than that in PN. Systolic blood pressure (odds ratio，OR 2.563)， urine protein excretion (OR 2.467)， uric acid (OR 2.323) and total cholesterol (OR 2.357) were independent risk factors for the disease progression by multivariate analysis. Conclusions HN diagnosed clinically without renal pathological evidence can not exclude primary nephritis. Further biopsy-based study is necessary to establish the true dimension of HN. High systolic blood pressure，urine protein excretion， total cholesterol and uric acid promote the progress of renal disease.

Objective To observe the effects of overexpressive Smad7 on the infiltration of inflammatory cells and the expression proinflammatory factors in peritoneum of rat peritoneal fibrotic model. Methods Forth-eight male SD rats were divided into four groups:normal control group； peritoneal fibrotic model group；empty vector group；Smad7 gene transfer group. All the rats with peritoneal fibrosis received daily intraperitoneal injection with 4.25% glucose dialysate(100 mg/kg BW) for 4 weeks and LPS(0.6 mg/kg BW)at day 8，10，12，22，24，26. Empty vector and Smad7 were transferred respectively with Tet-on/vector and Smad7 gene at day 0 and day 14 in empty vector group and Smad7 gene transfer group. All the rats were sacrificed at day 28 after peritoneal fibrosis model were induced.OX-1，ED-1 positive cells and IL-1 and TNF-α protein expression were measured by indirect immunoflurence.IL-1 and TNF-α mRNA expression were examined with RT-PCR. Smad7 gene transfer with ultrasound microbubble induced optimal gene transfection method.Results Comparing with the rats in peritoneal fibrotic and empty vector group， the infiltration of OX-1 and ED-1 positive cells in peritoneum was markedly improved in smad7 gene transfer group. IL-1 and TNF-α expression，including mRNA and protein， were also down regulated. Conclusion Overexpression of Smad7 with ultrasound microbubble induced gene therapy can successfully suppresse the infiltration of inflammatory cells and higher expression of IL-1 and TNF-α in rats with peritoneal fibrosis induced by high glucose and LPS.

Objective To investigate the effects of SPARC (secreated protein acidic and rich in cysteine) and its peptide on proliferation， apoptosis and cell cycle of human mesangial cells cultured in vitro， and explore the possible mechanism. Methods Mesangial cells were incubated in the media with various concentrations of SPARC and its peptide cultured in vitro. Cell proliferation was assessed by MTT colorimetric assay. Cell cycle and apoptosis index were analyzed by flow cytometry. The expression of cyclinD1 and p21Wafl proteins in response to SPARC and its peptide in HMC was determined by Western blot. Results Various concentrations of SPARC and its peptide could significantly inhibit the proliferation of mesangial cells in dose- and time-dependent manner， regulate the cell cycle at phrase G0/G1 increased while cells phrase S reduced，and could also induce apoptosis. Under the stimulation of SPARC and its peptide， the expression of cyclinD1 in HMC decreased markedly meanwhile the expression of p21Wafl increased significantly. Conclusions SPARC and its peptide can effectively inhibit HMC proliferation and regulate cell cycle progression. The mechanism may be mediated by inhibiting cyclinD1 and stimulating p21Wafl expression， subsequently blocking cells passing through G-S check point， which will be useful for treating mesangial proliferative glomerulonephritis.

Objective To investigate the relationship of up-regulated thrombospondin 1 (TSP-1) expression with renal interstitial fibrosis and peritubular capillary loss in chronic aristolochic acid nephropathy(CAAN). Methods Thirty-six male SD rats were randomly divided into two groups， 18 in each one. CAAN rats received 20 mg&#8226;kg-1&#8226;d-1 aristolochic acid (AA) contained in ethanol extract of Aristolochia manshuriensis Kom by gavage for 5 days in the 1st week， and after a break of 9 days， then 15 mg&#8226;kg-1&#8226;d-1 AA for 7 days in every other week up to the 12th week. Control group rats (Con) only received tap water by gavage as above. At the end of the 4th， 8th and 12th week， 6 rats in each group were sacrificed. Immunohistochemical staining was performed in rat renal tissue sections to examine the expression of TSP-1， transforming growth factor-β1 (TGF-β1)， aminopeptidase P(APP)， vascular endothelial growth factor (VEGF) and typeⅠcollagen (ColⅠ). Results Compared with Con， the expression levels of interstitial TSP-1， tubular TGF-β1 and interstitial ColⅠwere all up-regulated in CAAN rats (all P < 0.01)， meanwhile significantly positive correlations were found among them (TSP-1 vs TGF-β， r=0.925； TSP-1 vs ColⅠ，r=0.910； TGF-β1 vs ColⅠ r=0.857；all P < 0.01). Compared with Con， interstitial APP expression was significantly down-regulated in CAAN rats(P < 0.01)and correlated with interstitial TSP-1 and Col I expression(r=-0.945、-0.883，all P < 0.01)significantly and negatively. Compared with Con， tubular VEGF expression was significantly up-regulated in CAAN rats (P < 0.05 or < 0.01) and significantly correlated with TGF-β1(r=0.625， P < 0.01)and APP(r=-0.607， P < 0.01). Conclusions The up-regulated expression of TSP-1-TGF-β axis and the loss of peritubular capillaries related with TSP-1 may participate in the development of renal interstitial fibrosis in CAAN. The increased VEGF expression is probably a compensation phenomena.

Objective To evaluate the effect of genistein on phenotype of mesangial cell and extracellular matrix of diabetic mellitus (DM) rats. Methods The STZ-induced SD rats were divided into two groups: DM+Gen group receiving genistein intragastric admistration for 4 and 8 weeks and DM group without genistein administration for 4 and 8 weeks as well. Fast blood glucose(FBG)， BUN， Scr， SMemb mRNA， fibronetin (Fn)/kidney weight and MMP-2/TIMP-1 deposition were measured. Results As compared to control group， FBG， BUN ，Scr，Fn/kidney weight were all higher， especially at week 8(P < 0.01)；the immunohistochenistry semi-quantitative deposition of MMP-2/TIMP-1 decreased (P < 0.01)；the semi-quantitative ratio of SMemb mRNA/ GAPDH mRNA increased， peaked at week 4 and decreased at week 8 (0.794±0.037 vs 0.708±0.029)in DM group. Compared with the same time of DM group， genistein could ameliorate renal function partly， decrease SMemb mRNA/GAPDH mRNA ratio significantly (P < 0.01)； reduce Fn/kidney weight (P < 0.01)， and increase the deposition of MMP-2/TIMP-1 (P < 0.01 or < 0.05). Conclusions Genistein can down-regulate the expression of SMemb in renal cortex of STZ-induced DM rats， increase the deposition of MMP-2/TIMP-1， ameliorate renal disfunction of DM rats， thus， it may delay the progression of diabetic nephropathy.

Objective To study the relationship between the change of angiopoietins in glomeruli and loss of glomerular capillary in progressive glomerulosclerosis following podocyte injury. Methods One hundred male Wistar rats were randomly allocated into sham operation group (Sham，n=30)， uninephrectomy group (UNX，n=30) and uninephrectomy+daunorubicin group (DRB，n=40). The rats in DRB group were injected into rat`s tail with DRB(5 mg/kg) on day 7 and day 14 after UNX. At week 1，2，4，6 and 8 respectively following finishing animal model，6 rats in each group were taken randomly for determining creatinine cleareace(Ccr)，sections of kidneys were examined by PAS staining， immunohistochemical staining and in situ hybridization histochemistry for semiquantitative morphological evaluation. Glomerular cell apoptosis was examined by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL) and transmission electric microscope(TEM). Results In DRB groups， the magnitude of quantitative analysis in morphology shows that there was a decreasing trend of expression of Ang-1 mRNA and protein，but expression of Ang-2 mRNA and protein as well as expression of Fas，FasL and caspase-3 protein tended to increase，meanwhile glomerular apotopsis index(GAI)and glomerular sclerotic index(GSI)elevated with a decline in glomerular capillary density(GCD)and Ccr gradually. Apoptotic endothelial cells in glomeruli were found by TEM. Moreover， expression of Ang-1 mRNA and protein was negatively correlated with GAI， GSI，expression of Fas， FasL and caspase-3 protein， and a positively correlated with GCD and Ccr. Expression of Ang-2 mRNA and protein was positively correlated with GAI，GSI，expression of Fas， FasL and caspase-3 protein， and negatively correlatied with GCD，Ccr. Conclusion Podocyte injury may lead to an alteration in expression of Ang-1 and Ang-2 within glomeruli， which is correlated with up-regulating expression of Fas， FasL and caspase-3 protein， and facilitates the apoptosis of endothelial cell in glomeruli to result in the glomerular capillary loss relative to progressive glomerulosclerosis.

Objective To investigate the mechanism of early anemia in rat model of chronic aristolochic acid nephropathy(CAAN). Methods Seventy-eight female Wistar rats were randomly divided into three groups. Group A(n=30) were treated with caulis aristolochiae manshuriensis(CAM) decoction for 8 weeks. Group B (n=24) were treated with drinking water for 8 weeks. Group C were 5/6 nephrectomized rats (n=24). At week 8， 12 and 16， 1/3 rats in each group were sacrificed respectively. Before the rats were sacrificed， specimens of blood， urine and bone marrow were taken for biochemistry， ELISA， pathology， immunohistochemical analysis and the quantitation of erythropoietin (EPO). Results BUN and Scr of group A and group C increased at week 8， but the renal function of group C deteriorated quickly. Group A presented anaemia at week 12， and became worse at week 16. Hemoglobin (Hb) level of group A was lower as compared with group B and group C，P < 0.01. Compared with group B and group C， the TNF-α and IL-1β in blood serum and myeloid tissue of group A increased significantly(P < 0.01)， along with the decrease of CD34 expression and MVD. EPO level of each group was normal. Conclusion The expression of TNF-α and IL-1β increases at the early phase of CAAN， and the microvessel of bone marrow is markedly injured， which may result in early onset of severe anemia in rat CAAN model.

Objective To detect the levels of renin and angiotensinⅡ in renal tissue and plasma from kidney transplantation patients suffering from chronic CsA-related nephropathy，and to explore the effect of renin-angiotensin system (RAS) on chronic cyclosporine nephropathy. Methods (1)Thirteen patients with renal biopsy-proven CsA-related nephrotoxicity were enrolled in this study， whose renal tissues were subjected to immunohistochemical staining by rabbit polyclonal anti-human renin antibody. Cyclosporine plasma level was obtained 2 hour after morning dose (C2) and angiotensin Ⅱ (AngⅡ) plasma level was measured after allograft biopsy was performed. The relationship between renin expression and clinicopathological manifestation was also investigated.(2)Human umbilical vascular endothelial cells(HUVEC)and mesangial cells(MC)were incubated with different concentrations of CsA (0，250，500，1000 μg/L) for 24 hours. Immunohistochemical staining was used to examine renin expression. The secretion of AngⅡ in cytoplasm and culture medium was measured by radioimmunoassay. Results The histological renin index was significantly higher in specimens of chronic CsA nephropathy than that of control (5.20±0.40 vs. 2.30±0.24，P < 0.01). The plasma levels of C2 and AngⅡ in chronic CsA nephropathy group were higher than those in control group，but no significant difference was found[(122.69±26.73)pg/ml vs.(121.88±36.35)pg/ml，P=0.977，(719.04±55.89)pg/ml vs.(658.80±90.78)pg/ml，P=0.196，respectively].In vitro， renin expression increased significantly both in HUVEC and MC after the cells were incubated with CsA for 24 hours(231.50±12.10 vs.182.34±11.51，P < 0.05； 176.35±14.51 vs.153.36±11.11，P < 0.05). CsA also stimulated the secretion of AngⅡ in HUVEC and MC in a dose-dependent manner. Conclusions Renal allograft biopsy is important to differentiate chronic CsA-related nephropathy from chronic rejection. The intrarenal RAS may play an important role in chronic CsA-related nephropathy. The histological lesion of CsA nephrotoxicity fails to correspond spontaneously to either the change of C2 level or the change of AngⅡ plasma level. CsA stimulates the secretion of AngⅡ and the expression of renin in HUVEC and MC. Blockage of RAS may be helpful for therapeutic intervention in the progression of chronic CsA-related nephropathy.