Objective To study the cytopathogenic effect of epidemic hemorrhagic fever with renal syndrome virus (HFRSV) on renal tubular cells(RTC). Methods Human fetal renal tubular cells (HFRTC) were cultured in vitro. HFRTC infected or not infected by HFRSV were observed by using trypan-blue stain and transmission electron microscopy(TEM). Viral-mRNA was detected by in situ molecular hybridization. Results (1) HFRSV could directly infected HFRTC: (2)The death rate of HFRTC in the infection group was significantly higher than that in the control grou 1 to 4 weeks after infection; (3) Injuries of cell membrane and cell organs after infection with HFRSV were significantly earlier and more severe as compared to control by means of TEM. Conclusion HFRSV can directly damage renal tubular cells (RTC ), which contributes to the pathogenesis of hemorrhagic fever with renal syndrome (HFRS).

Objective To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cells (HKC).Methods Cultured HKC were divided into 3 groups: (1) negative control; (2) low dose CTGF treated (rhCTGF 2. 5 ng/ml); (3) high dose CTGF treated (rhCTGF,5. 0 ng/ml). Expression of ?-smooth muscle actin (?-SMA) and fibronectin(FN) mRNA were measured by RT-PCR. Indirect immunofluorescence and flow cytometry methods were used to assess the level of intracellular ?-SMA protein. Concentration of FN secreted into the media was determined by ELBA. Results Upon the stimulations of different concentrations of rhCTGF,expression of ?-SMA and FN mRNA increased markedly(P

To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of alpha-smooth muscle actin (alpha-SMA) were assessed by indirect immuno-fluorescence, and the percentage of alpha-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of alpha-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of alpha-SMA were markedly stronger than that in negative controls. The percentages of alpha-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9%, 65.5% vs 2.4%, P<0.01). alpha-SMA mRNA levels were also up-regulated by the stimulation of rhCTGF (P<0.01). These results suggest that CTGF can promote the transdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).
Actins/metabolism ; Cell Differentiation/*drug effects ; Cells, Cultured ; Epithelial Cells/*cytology ; Immediate-Early Proteins/*pharmacology ; Insulin-Like Growth Factor Binding Proteins/pharmacology ; Intercellular Signaling Peptides and Proteins/*pharmacology ; Kidney Tubules/*cytology

Objective To study the effects of CD2-associated protein (CD2AP) on podocyte adhesion and extension ability and to explore its possible mechanism. Methods Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 medium at 33℃permissive conditions. The podocytes were transfected with CD2AP small interfering RNA (siRNA) and serambing sequences labeled with fluorescein were taken as control. The transfected podocytes were trypsinized and seed into collagen IV coated plates. The relative cell adhesion and cell area were examined 90 min later. Apoptotic rates of CD2AP siRNA transfected podoeytes and different PAN concentrations incubated podoeytes were detected by flow cytometer. The distribution of F-actin was observed under laser scanning confoeal microscope. Nephrin protein expression and its phosphorylation level were examined by immunofluorescence and Western blot. Results The relative ceil adhesion of CD2AP siRNA transfected podocytes was apparently lower than that of control group[(41.72±6.07)% vs (64.46±8.53)%, P<0.05]. The cell area analysis had the similar result. The apoptotic rate of CD2AP siRNA transfected podocytes was significantly higher than that of the controls [(5.73±0.61)% vs (3.26±0.45)%, P<0.05]. 100 mg/L PAN could markedly induce podocytes to apoptosis and impair cell adhesion ability (P<0.05). Nevertheless, no significant difference was found in cell body spreading (P>0.05). The distribution of F-actin in CD2AP depletion podocytes was apparently altered. The expression of nephrin protein and its phosphorylation level was conspicuously descended to some degree (P<0.05). Conclusions CD2AP depletion facilitates podocyte apoptosis and impairs cell adhesion function. Cytoskeleton confusion and nephrin signaling weakness caused by CD2AP depletion may he partly responsible for the decline of cell adhesion and spreading.

Objective To establish and optimize the two-dimensional gel electrophoresis technical platform for the blood serum proteome research in patients with end stage renal disease(ESRD).Methods Immobiline pH gradients isoelectric focusing was used as the first dimensional gel electrophoresis and the vertical SDS-PAGE was used as the second dimensional gel electrophoresis(2-DE).The 4 differentially expressed protein spots were identified by mass spectrometry.Results Satisfactory 2-DE maps of ESRD patients serum protein were obtained and there were some differentially protein spots between the 2-DE maps of ESRD patients and normal controls.Conclusions The 2-DE technology for the serum proteome of ESRD patients is set up.

AIM:To study the potential pathological role of abnormal expression of endogenous angiopoietins in progressive glomerulosclerosis.METHODS:80 male Wistar rats were randomly allocated into sham operation group(sham,n=25),unilateral nephrectomy group(UNx,n=25)and UNx+daunorubicin(DRB)group(n=30).The rats in DRB group were intravenously injected with DRB(5 mg/kg)on the seventh and the fourteenth day respectively after excising one kidney.Then,at week 1,2,4,6 and 8,5,male Wistar rats from each group were taken randomly for determining 24 h urinary protein quantitative measurement(24hUPQ),BUN,Scr,and the kidneys were examined by electronic microscope,PAS staining,immunohistochemical staining and in situ hybridization histochemistry.RESULTS:There was a trend towards an increase respectively in levels of 24hUPQ,Bun,Scr,GSI in DRB from week 2 to week 8.Electronic microscope revealed that podocyte injury presented in DRB group.Expression of Ang1 mRNA and protein in glomerulus in DRB group decreased,while expression of Ang2 protein in glomeruli in DRB group increased.In DRB group,expression of Ang1 protein had a negative correlation with 24hUPQ,BUN,Scr,GSI,expression of Ang2 protein and CoIV protein.Expression of Ang2 protein had a positive correlation with 24hUPQ,BUN,Scr,GSI,expression of CoIV protein.CONCLUSION:Podocyte injury may lead to glomeruli abnormally express angiopoietins.A decrease in expression of Ang1,and upregulation in expression of Ang2 may facilitate progressive glomerulosclerosis in the rat.

AIM: To establish a model of subtotal nephrectomy (SNx) and investigate the changes of apoptosis and apoptosis-related genes (Bax, bcl-2, caspase-3, caspase-8 and caspase-9) in the rat remnant kidney. METHODS: Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at 1, 2, 4, 8, 12, 16, 26 and 40 weeks after operation. The tissues of remnant kidneys were collected to detect apoptosis cells by in situ end-labeling of cleaved DNA (TUNEL) and proliferating cells by determining the proliferating cell nuclear antigen (PCNA). The expression of Bax, bcl-2, caspase-3, caspase-8 and caspase-9 was measured by RT-PCR and Western blotting. The proteins were detected by immunohistochemistry staining. The relation between apoptosis, proliferation, glomerulosclerosis and renal interstitial fibrosis was also observed. RESULTS: The results showed the renal pathological dynamic changes in 5/6 nephrectomy remnant kidneys were tubule-interstitial inflammation and fibrosis, as well as glomerulosclerosis. There were transient increases in both proliferating and apoptotic processes in glomerulus, tubules and interstitium. Apoptosis was increased and most of apoptotic cells were detected in tubular epithelial cells and interstitial area. The mRNA and protein expression of Bax, caspase-3, caspase-8 and caspase-9 were increased in all course, and peaked at week 4 and 40 in the SNX rats. The successive changes of these parameters were parallel to the level of focal inflammation in interstitium. Glomerulosclerosis index was related with focal inflammation cells and 24 hours urine protein (r=0.788, 0.822; P

AIM:To observe the expression of connective tissue growth factor(CTGF)in unilateral ureteral obstruction(UUO)rats,and to explore its pathogenic role in renal tubulointerstitial fibrosis.METHODS:48 Wistar rats were randomly divided into sham-operated and UUO group.The rats were sacrificed at day 1,3,7,and 14.The degree of tubulointerstitial damage was scored according to the Masson staining.The mRNA and protein levels of CTGF,transforming growth factor-?1(TGF-?1),collagen Ⅰ(Col Ⅰ),and plasminogen activator inhibitor-1(PAI-1)were detected by reverse transcriptional-polymerase chain reaction(RT-PCR)and immunohistochemistry,respectively.Expression of CTGF protein in the kidney was also assessed using Western blotting.RESULTS:TGF-?1 mRNA level began to increase as early as 1 day after UUO.This increase was followed by the elevation of CTGF mRNA level,which began to increase at third days after UUO(P

To investigate the effects of mycophenolate mofetil (MMF) on the process of renal interstitial fibrosis, unilateral ureteral obstruction (UUO) model was established in rats. Twenty Sprague-Dawley rats underwent UUO and received vehicle (n = 10) or MMF (20 mg.kg-1.d-1, by daily gastric gavage, n = 10) during a period of 5 days following surgery, and the additional 10 rats were served as sham-operated group. The rats were killed 5 days after surgery. Immunohistochemistry was performed on renal tissue for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA) and type I and III collagen (col I, col III). Histological studies were also done by MASSON staining. Five days after surgery, proliferating cells in tubules, interstitium as well as interstitial myofibroblast (MyoF) infiltration and interstitial col I, col III deposition were all significantly reduced by MMF treatment. MMF also alleviated the histological changes of UUO rats. These results suggested that the reduction of interstitial MyoF infiltration may be an important event by which MMF prevents renal injury caused by UUO and MMF could be used to limit the progression of renal fibrosis.
Fibrosis ; Kidney/*pathology ; Kidney Diseases/etiology ; Kidney Diseases/pathology ; Kidney Diseases/*prevention & control ; Mycophenolic Acid/*analogs & derivatives ; Mycophenolic Acid/*pharmacology ; Random Allocation ; Rats, Sprague-Dawley ; Ureteral Obstruction/*complications

Objective To investigate the fungal infection situation and the risk factors in lupus nephritis(LN)patients.Methods Infection states were investigated in 93 LN patients and 45episodes(about 48.4%) occurred the different kind and different degree of Hospital-acquired infections.The fungal infection rate was 22.6% and two patients directly died of it.Except fungi,the main pathogens were bacteria and virus.The main infection sites were urinary,gastrointestinal and respiratory tract.Results The infection causes were related to the use of corticosteroids,immunodepressive,antibiotic and the aggressive operation,renal failure ,the duration in hospital and so on.Conclusion Effective prevention and treatment for infection in LN patients is an important way to improve the prognosis of LN.