Objective To study the expression of basic fibroblast growth factor(bFGF)in the kidney tissues of chronic interstitial nephritis(CIN).Methods Expression of bFGF was assayed in 30 patients with CIN and 5 normal persons by SP method of immunohistochemical technique.The average gray value and average optical density of the bFGF positive parts and its ratio with the whole visual field had been measured with video analysis system.Results The expression of bFGF in the kidney tissues of CIN was significantly stronger than that in normal tissues.The average gray value,average optical density and the positive areas were significantly high compared with that of normal cases.Conclusion The bFGF can promote the proliferation of fibroblasts and monocytes in renal interstitial patients and lead to renal interstitial fibrosis in the end.

Objective To investigate the mechanism of immunointerstitial nephritis. Methods BN rat model was obtained with bovine tubular base membrane (bTBM). Monoclonal antibodies, immunohistochemical ABC method and indirect im-munofluorescent technique were used to detect the inflammatory cells and ICAM-1. Results 9 days after immunization with bTBM, inflammatory cells infiltration and a strong ICAM-1 expression were found in the intersitium, associated with the pathological degree. Conclusion Cellular immunity mediated by ICAM-1 may play an important role in the development of immunointerstitial nephritis. Macrophage is a leading factor among various inflammatory cells. Fluid immunity can not be excluded.

Objective To observe the effects of calcium dobesilate on the expression of CD34 and VOH Willebrand factor(vWF)in peritubular capillary (PTC)in renal tissues of rats with chronic arisolochic acid nephropathy(CAAN)and to explore the probable mechanism concerned.Methotis Sixteen Wistar rats were infused with Caulis aristolochia manshuriensis decoction for 12weeks,then randomly divided into 2 groups.NX group rats(n=8)were infused with distilled water and treatment group(n=8)with calcium dobesilate at the following 4 weeks solely.Control group rats(n=8)were infused with distilled water for the 16 weeks.At week 16,all the rats were sacrificed.Specimens of blood and urine were collected to detect the blood urea nitrogen (BUN),serum creatinie(Scr)and urine protein.HE and Masson staining was used to observe the pathology of the kidney.Immunohistochemistry was used to detect the expression of CD34 and vWF.Results Urinary protein,Scr and BUN in calcium dobesilate treatment group were much lower than those in NX group (P＜0.05).The A value of CD34+ increased significantly in calcium dobesilate treatment group[(16.72±4.17)×103]compared with NX group[(3.19±1.40)×103]at week 16(P＜0.01).The A value of vWF+decreased in calcium dobesilate treatment group[(10.16±1.68)X103]compared with NX group[(18.66±4.65)x103]at week 16(P＜0.01). Conclusion Calcium dobesilate can increase the expression of CD34 and the density of peritubular capillary(PTC)in renal tissues of CAAN rats,and reduce the expression of vWF and the formation of microthrombosis.

Objective To investigate the effects of angiotensin receptor blocker (ARB)losartan on the glomerular protein expression profile of spontaneous type 2 diabetic KKAy mice by two-dimensional differential gel eleetrophoresis and MALDI-TOF mass spectrometry.Methods 8-week-old spontaneous type 2 diabetic KKAy mice were randomly divided into losartan (10 mg·kg-1·d-1 given in drinking water) treatment group and non-treatment group.Eight-week-old C57BL/6 mice were used as normal control.The glomeruli were separated by magnetic bead perfusion through thoracic aorta at age of 20 weeks,then glomerular protein was extracted.The glomerular protein expression profile was investigated by CyDyes minimal fluorescence labelling,two-dimensional differential gel electrophoresis and MALDI-TOF mass spectrometry.Results KKAy mice developed higher body weight and blood glucose,higher urinary microalbumin creatinine ratio at age of 20 weeks than C57BL/6 mice at the same age (all P＜0.05).Losartan treatment markedly reduced urinary microalbumin creatinine ratio [(539.71 ±100.23)mg/g vs (728±177.19) mg/g],attenuated mesangial expansion and the thickening of glomerular basement membrane,but had no effect on the blood glucose.By DeCyder 2-D differential analysis software,62 protein spots of differential expression were found in glomeruli between losartan treatment and non-treatment KKAy mice at age of 20 weeks.Among them,41 proteins were identified by peptide mass fingerprinting.The expressions of 28 proteins were up-regulated by losartan treatment,including glycerokinase,sulfite oxidase,glycine amidinotransferase,adenosylhomocysteinase,etc.The expressions of 13proteins were down-regulaled by losartan treatment,including 3-mercaptopyruvate sulfurtransferase,ATP synthase subunit d,60 000 heat shock protein,stress-70 protein (alternative name 75 000glucose-regulated protein,GRP75),etc.Six differcntially expressed proteins were found in glomeruli between non-treatment KKAy mice and C57BL/6 mice,and the differential expressions were suppressed by losartan treatment,including dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex,succinyl-CoA ligase (GDP-forming) subunit beta,mitochondrial,ATP synthase subunit d,GRP75,nucleoside diphosphate-linked moiety X motif 19 and seleniumbinding protein 1.Conclusions Losartan significantly reduces the urinary protein excretion rate and renal pathological lesion of spontaneous type 2 diabetic KKAy mice,and suppresses the differential expression of mitochondrial ATP synthase subunit d,GRP75,selenium-binding protein 1,etc in glomeruli.Losartan may play a renoproteetive role by reducing glomerular mitochondrial reactive oxygen species genesis and inhibiting oxidative stress.

ObjectiveTo observe the therapeutic effects of Shenxiong glucose injection in the patients with chronic kidney diseases.Methods Seventy-eight patients with chronic kidney disease were given intravenous glucose injection 200 ml,qd,for 2 weeks.The patients who were complicated with diabetes would be given insulin 2 U/100 ml.Before and after Shenxiong injection treatment,24 h urinary protein,blood urea nitrogen ( BUN),creatinine ( Cr),hemoglobin ( Hb),albumin ( ALB),hematocrit ( HCT),fiber fibrinogen (FIB),cholesterol (TC) and triglyceride (TG) were measured and compared.Results After 1 course of Shenxiong treatment,the serum BUN,Cr,FIB,24 h urinary protein,Hb and HCT ( [ 15.70 ± 3.62 ] mmol/L vs.[6.74 ± 1.56 ] mmol/L,[ 564 ± 65 ] μmol/Lvs.[ 189 ± 43 ] μmol/L,[ 0.08 ± 0.01 ] g/L vs.[ 0.04 ± 0.01 ] g/L,[7.96 ±3.45]g vs.[3.60 ± 1.92]g,[83.6 ±10.5]g/L vs.[79.5 ±8.7]g/L,[0.43 ±0.0] vs.[0.39 ±0.06 ] were decreased,and ALB ( 28.7 ± 8.6) vs.( 36.8 ± 6.2) was increased.The difference was statistically significant (t =3.1 1,2.98,3.04,2.82,2.02,2.23,P＜0.01 for all).TC and TG were not changed much after the treatment.ConclusionShenxiong injection can improve the kidney function,lower Fibrinogen and urine protein,which may effectively slow down the progress of chronic kidney disease and improve the life quality.

ObjectiveWith a GcneChip(R) cxpression analysis,98 known gencs and 31 exprcssed sequence tags (ESTs) were found to be differentially expressed between KK/Ta and BALB/c kidneys.To further screen the susceptibility genes for diabetic nephropathy,the temporal and spatial expression patterns of differentially expressed gene-adipsin were investigated.MethodsThe body weight,blood glucose,urinary albumin/creatinine ratio,and renal pathological changes of KK/Ta and BALB/c mice were measured at the 7,20,28 and 36 weeks of age.Total RNA was extracted from the kidney,heart,liver,lung,and brain.The temporal and spatial expression patterns of adipsin in diabetic KK/Ta mice were examined by competitive RT-PCR.The correlation analysis between adipsin expression and albuminuria level was carried out.ResultsThe mRNA expression of adipsin was found in the kidney,heart,lung,and brain,but not in liver.The expression of adipsin in diabetic KK/Ta mice at 20 weeks of age was significantly down-regulated in kidney,heart,and lung than that in age-matched BALB/c mice,and unaltered in brain.Adipsin expression in KK/Ta kidneys was significantly down-regulated with aging and negatively correlated to urinary albumin/creatinine ratio( r =-0.807,P ＜ 0.05).ConclusionsThe expression of adipsin mRNA was downregulated in kidney,heart,and lung in diabetic state.Adipsin expression in KK/Ta kidneys was negatively correlated to urinary albumin/creatinine ratio.It might be a candidate gene for diabetic nephropathy.

Objective A total of 102 ESRD patients undergoing hemodialysis for over 3 months in our dialysis center were asked to complete the SF-36 scales.They were also assessed by Hamilton Depression Scale(HAMD17).Univariate analysis was performed to determine the impact of factors such as age,gender,employment status,education,mental status and dialysis status on their quality of life.Methods The 102 patients with ESRD undergoing hemodialysis for over 3 months in our dialysis center completed the SF-36 scales with self-administration,and they were also assessed by Hamilton Depression Scale (HAMD17).Univariate analysis was performed to determine the impact of variables such as age,gender,employment status,education,mental status and dialysis status on the quality of life in patients.Results The scores of PF,RP,BP,GH,VT,SF,RE and MH in patients were significantly lower than those in the normal controls(P

Objective .To prove aristolochic (AA) caused vascular endothelial cells (VEC) injury via intracellular calcium overloa-ding and investigate the mechanism of calcium dobesilate antagonism. Methods Human umbilical vein endothelial cells (HUVEC) were cultured in vitro, and randomly divided into three groups: Control group, AA group, intervention group. Microscope and transmission elec-tron microscopy were used to observe changes of cell morphology and ultrastructure. ELISA method were applied to determine thrombomedu-lin (TM) in cell culture supernatant, fluorescent indicator FLuo-3/AM and intracellular calcium concentration ([Ca2+]. Results TM val-ue and average [Ca2+] i of AA group were significantly higher than that of control group (P < 0.05). Compared with the AA group, when the concentration of calcium dobesilate was 25 μM or 50 μM, TM value and average [Caz +] significantly decreased in intervention group (P < 0.05). Compared with control group, endoplasmic reticulum was pool expansion shaped, and mitochondrial cristae was absent in AA group cells. Endoplasmic reticulum and mitochondria patterns in the intervention group cells showed some improvement, compared with AA group. Conclusion AA induced VEC calcium overloading, 'I'M secretion and injury of endothelial ceils, endoplasmic reticulum and mito-chondria destruction. Dabesilate calcium could protect endoplasmic reticulum and mitochondria and reduce AA induced VEC calcium over-loading, and these could protect VEC.

Objective To investigate the renoprotection of tubular L-FABP in murine IgA nephropathy (IgAN) induced by bone marrow transplantation(BMT). Methods IgAN models were reconstituted by BMT from IgAN-prone mice into mice (Tg) transgenically tubular overexpressing human L-FABP (hL-FABP) and wild type (WT) mice. These recipients were sacrificed at 6 and 12 weeks after BMT and their kidneys were collected. The expressions of hL-FABP, fibronectin (FN)and monocyte chemoattractant protein-1 (MCP-1) mRNA were detected by real-time PCR. hL-FABP,FN, type Ⅳ collagen (Col Ⅳ ), hemeoxygenase-1 (HO-1) and 4-hydroxy-2-nonenal (4-HNE)modified proteins were detected by Western blotting. The distribution of hL-FABP and FN protein in kidney was detected by immunohistochemistry. The level of serum IgA, urinary albumin and urinary hL-FABP was detected by ELISA. Results (1) IgAN was reconstituted in both Tg and WT mice by BMT: mesangial IgA deposition and up-regulation of serum IgA. The levels were not significantly different between two groups (Tg-ddY and WT-ddY). (2) hL-FABP was expressed in proximal tubular cells of normal Tg mice. The mRNA (1.62±0.32 vs 0.46±0.09, P＜0.01) and protein expression (1.74±0.76 vs 1.14±0.31, P＜0.01) of hL-FABP was up-regulated in Tg-ddY kidney and urinary hL-FABP level (μg/g creatinine) was significantly increased (59.87±26.75 vs 31.01±14.86, P＜0.05) at the 6th week after BMT. (3) WT-ddY mice showed a significantly higher urinary albumin level (mg/L) (828±656 vs 82±22, P＜0.01), severer mesangial matrix expansion (P＜0.01),more glomerular FN and Col Ⅳ deposition at the 12th week. (4) Up-regulation of renal hL-FABP was associated with significant suppression of renal HO-1 expression (P ＜0.05),accumulation of 4-HNE modified proteins (P＜0.05) and MCP-1 mRNA expression (P＜0.01) in Tg-ddY mice. Conclusion Tubular L-FABP may lessen the progression of glomerular damage at early stages of IgAN by reducing oxidative stress and inflammatory mediators.

Objective To identify the candidate genes in the vicinity of a susceptibility locus (urinary albumin 1,UA-1) contributing to the development of albuminuria in type 2 diabetic KK/Ta mice. Methods Total RNA was extracted from the kidneys of KK/Ta (n=3) and BALB/c (n=2) mice at 20 weeks of age.The gene expression profile in kidney was investigated using the Affymetrix Murine Genome U74Av2 array.Competitive RT-PCR was used to confirm the differential expression of syndecan-4 which located in the vicinity of UA-1.Genome DNA was extracted from KK/Ta and BALB/c mice.DNA sequence analysis of the coding and promotor region of syndecan-4 gene was conducted. Results In the vicinity of the susceptibility locus (UA-1)contributing to the development of albuminuria in type 2 diabetic KK/Ta mice,10 candidate genes that showed differential expression were identified.Among them,the gene expression of syndecan-4in KK/Ta kidneys at 20 weeks of age was up-regulated by 26.1 times of age-matched BALB/c kidneys.Sequence analysis revealed two synonymous polymorphisms in the coding region (A93C and T216C) and three polymorphisms in the promoter region (-T263C,-T396C and -G669A) of the syndecan-4 gene.The TATA box was found at 321 bp upstream from the transcription start site,and the T263C polymorphism was located in the binding site of transcription factor Clox.Conclusions Syndecan-4 gene is mapped in the vicinity of the susceptibility locus contributing to the development of albuminuria in type 2 diabetes.The gene expression of syndecan-4 in KK/Ta kidneys is up-regulated than that in age-matched BALB/c kidneys at 20 weeks of age.Thus syndecan-4 may be one of the potential candidate genes responsible for diabetic nephropathy.Sequence differences in the promoter region influence the expression levels of syndecan-4 genes in KK/Ta kidneys.