[Objective] To investigate the therapeutic effect of Honghua injection plus prednisone and cyclophosphamine on refractory nephrotic syndrom.[Methods] 38 patients with refractory nephrotic syndrom were given intravenous Honghua and cyclophosphamine plus oral prednisone.Serum total cholesterol,triglyceride,liver and renal function,blood routine test,24h protennuria were observed.[Results] Total effective ratio was 92.1%.Cure ratio was 73.68%.Partly cure ratio was 18.42% and ineffective ratio was 7.89%.[Conclusions] Patients with refractory nephrotic syndrom were treated by Honghua injection plus prednisone and cyclophosphamine,which could reduce the application of prednisone,its side effects and enforce its therapeutic effect.What is more,this treatment could lessen therapeutic process,prevent relapse and recoil,protect effectively the retreat syndrom of prednisone as well.

Objective:To explore the role of osteopotin in lupus nephritis (LN).Methods:Sera osteopotin( OPN )levels were measured by enzyme linked immunosorbent assay in 90 patients with systemic lupus erythematosus (SLE),15 non-SLE patients with renal impairment,and 30 healthy volunteers.OPN mRNA expression in peripheral blood mononuclear cells (PBMCs) was also investigated with reverse transcription-polymerase chain reaction semi-quantitative method.Results:The expression of OPN was significantly higher in active LN groups than in all other groups(P0.05).Conclusion:The expression of PBMCs OPN mRNA is up regulation in active SLE.Meanwhile,its expression levels are correlated with the activity of LN.

Objective To investigate the effects of the combination of tripterygium wilfordii Hook F (TwHF) and irbesartan on urinary podocyte in diabetic kidney disease (DKD) patients,and to discuss the mechanism of protective effect of TwHF on DKD.Methods A total of 45 type 2 diabetic kidney disease patients were enrolled into this prospective study,and were randomly divided into 3 groups:TwHF treatment group (DT,n =15),irbesartan treatment group (DI,n =15),and TwHF combined with irbesartan treatment group (DTI,n =15).After 6 weeks washout,the 3 groups were given TwHF (1-2 mg · kg-1 ·d-1),irbesartan (150-300 mg/d),and TwHF (1-2 mg · kg-1 · d-1) combined with irbesartan (150-300 mg/d) for 12 weeks respectively.Fifteen healthy volunteers served as controls.Urinary podocytes were identified and quantitated by immunofluorescence staining of urinary sediments labeled by monoclonal antibody podocalyxin.In addition,we studied urinary connective tissue growth factor (CTGF),osteopontin (OPN) and transforming growth factor β1 (TGFβ1) concentrations in DKD patients by enzyme-linked immunosorbent assay.Results Urinary detached podocytes were obviously higher in the urine of DKD patients than in healthy controls (P ＜0.01).Podocyte detection rate was 86.6％ in the urine of DKD patients.The protein expressions of CTGF,OPN and TGFβ1 in patients with urinary podocyte were significantly increased than those without urinary podocyte (P ＜ 0.05 or 0.01).Correlation analysis showed that there was positive correlation between urinary protein excretion and urinary podocytes (r =0.79,P ＜ 0.01) and there were positive correlations between the number of urinary podocytes and urinary protein expressions of CTGF,OPN and TGFβ1 (r =0.56,0.41,0.44,respectively,all P values ＜ 0.01).Urinary albumin excretion and urinary podocytes were significantly decreased in all treatment groups (P ＜ 0.01),simultaneously,urinary concentrations of CTGF,OPN and TGFβ1 were reduced in all groups at week 12 after intervention of TwHF,irbesartan and TwHF combined with irbesartan (P ＜ 0.01),and these changes were more distinguished in combined treatment group (P ＜ 0.05).Conclusion Urinary podocyte in the urine may be suggested to be an early effective marker of disease activity in DKD.TwHF may be effective to prevent podocyte injury in DKD,which may be mediated,at least partly,by down-regulating the expression of CTGF,OPN and TGFβ1.There is a synergistic protective effect of TwHF combined with irbesartan on podocyte injury in DKD patients.

Objective To detect the effect and mechanism of Cyr61 on the apoptosis of renal tissue caused by early stage of ischemic acute kidney injury (AKI). Methods 30 SD rats were randomized into 5 groups, including control group, AKI group, AKI+bicarbonate group, AKI+blank virus group, and AKI+over?expression Cyr61 virus group. After animal models were created for 2h, serum and renal tissue were collected from sacrificed animals. Expression level of TNF?α was determined by ELISA. HE staining was used to observe the histologic changes of renal tissues. The levels of NF?κB p65 and TNFR1 were measured by immunohistochemical method. RT?PCR and Western blotting assay were adopted to detect the mRNA and protein expression levels of NF?κB p65, TNFR1 and Caspase3. Results Compared with control group, AKI group, AKI+bicarbonate group, AKI+blank virus group, AKI+over?expression Cyr61 virus group had obvious kidney injury. The levels of TNF?α, the mRNA and protein expression levels of NF?κB p65, TNFR1 and caspase3 were markedly up?regulated. Over?expression of Cyr61 significantly attenuated the degree of pathological injury, numbers of apoptotic renal tubular epithelial cells and increased the degree of Scr. Although compared with other groups, the level of TNF?α in kidney tissue had no difference, there was obvious decreased protein level of NF?κB p65, while the increase of TNFR1 and Caspase3 protein was moderate. Conclusions During the early stage of AKI, over expression of Cyr61 could inhibit apoptosis, which may be related to the suppression of TNFR1 transcriptional expression and interference of TNF?αpathway. Its underlying mechanism therefore deserves further research.

Objective To investigate the relationship between urine neutrophil gelatinase-associated lipocalin (uNGAL) level and activity and pathological types of lupus nephritis (LN).Methods Thirty cases of biopsy proven LN patients as the initial onset were enrolled into the study.Ten healthy persons were selected as controls.The clinical and pathological data and blood, urine specimen were collected.The uNGAL was measured by enzyme linked immunosorbent assay.The relationship between uNGAL and clinical and pathological features of LN was analyzed.One-way analysis of variance (ANOVA) and Pearson's correlation analysis were used for statistical analysis.Results Nineteen cases were in the LN active group and 11 cases were in the inactive group.The level of plasma NGAL had no significant difference between the active LN group [(64±6) ng/ml] and the inactive LN group [(58±20) ng/ml] and the healthy control group [(57±20) ng/ml] (P＞0.05).The level of urine NGAL in the active LN group [(69±3) ng/ml] and inactive LN group [(66±5) ng/ml] was higher than that in the healthy control group [(64±5) ng/ml, P=0.009, 0.016, respectively].Urine NGAL level in active LN group was higher than that in the inactive group (P=0.012).Urine NGAL level was positively correlated with SLEDAI, R-SLEDAI, GAI, TLAI, AI (r=0.472, 0.521, 0.502, 0.516, 0.597, respectively, P=0.042, 0.036,0.042, 0.021, 0.007, respectively).The urine NGAL concentration after comparing to different pathological conditions were: urine NGAL's level [(69.7±2.4) ng/ml] of type Ⅳ LN was higher than type ⅢN [(65.3±3.2)ng/ml] and type Ⅴ [(64.6±5.0) ng/ml] (P=0.031, 0.028, respectively).Receiver operating characteristic (ROC) indicated that uNGAL was more sensitive(86.7％) and specific (73.3％) for the diagnose of type Ⅳ LN than type Ⅲ or type Ⅴ LN.Conclusion uNGAL is closely related with disease activity and pathological activity in LN.uNGAL enables clinician to assess the activity of LN and could be a sensitive marker for the diagnosis of type Ⅳ adult LN.

Objective To investigate the relationship between the infiltration of lymphocytes in renal tissues and podocytes injury in patients with lupus nephritis (LN),and provide the evidence of mechanism of podocytes injury in LN.Methods Thirty cases of biopsy proven LN patients were enrolled into the study,10 cases were selected as a the controls.The clinical and pathological data and renal tissues were collected.The infiltration of T lymphocytes (CD4+,CD8+ cells) and B lymphocytes (CD20+ cells) in renal tissues were detected by immunohistochemical method.The nephrin expression was detected by immunofluorescence.The relationship between CD4+,CD8+,CD20+ cells in renal tissues and 24 h UP,nephrin expression were analyzed by Pearson's correlation analysis,respectively.Results ① Compared with the control group,the 24 h UP was increased [2.86±1.37 vs 0.10±0.22 (g/24 h)];the infiltrations of inflammatory cells were increased significantly,the podocytes injury could be observed,combined with effacement of podocytes foot processes or disappeared in the LN group.Moreover,the increase of 24 h UP of active LN group was more evident than that in the inactive group [3.91 ±1.45 vs 1.77±0.69 (g/24 h),F=24.15,P＜0.05],and the infiltration of inflammatory cellsand effacement of podocytes foot processes were more severe in the active LN group.②Compared with the control group,the nephrin expression decreased and the infiltration of CD4+,CD8+ and CD20+ cells in the renal tissues increased significantly,which was mainly aggregated in renal interstitial tissue.And In addition,CD4+ [98±13 vs 40±12 (cells/glomerulus),F=240.18,P＜0.05],CD8+[109.0±16.4 vs 53.3±12.1(cells/glomerulus),F=210.40,P＜0.05] and CD20+ [149.4±1.4 vs 82.6±13.3 (cells/glomerulus),F=544.30,P＜0.05] cells of the active LN group were more remarkable than that in the inactive group.③ The Pearson correlation analysis showed that CD4+,CD8+ and CD20+ cells in renal tissues were positively correlated with 24 h UP (r=0.688,0.748,0.702;P＜0.01),and were negatively correlated with nephrin expression (r=-0.623,-0.793,-0.693;P＜0.01).Conclusion The infiltrations of CD4+,CD8+ and CD20+cells in renal tissues are very closely related with podocytes injury in patients with LN,and the infiltration of lymphocytes may be an important mechanism for podocytes injury in LN patients.

Objective To investigate the renal expression of stem cell factor (SCF) in lupus nephritis (LN) and its correlation with disease activity and renal injury parameters. Methods Histochemical stain was used to examine all renal specimens (LN group n=34, chronic glomerulonephritis n=16, control group n=8). Hyhridization in situ and immunohistochemistry were used to detect the expression of SCF and infiltration of mast cells, macrophages , α-SMA (+) cells in renal tissues of the two groups. SPS software was used for tissue of the control group. However, they increased markedly in lupus nephritis and CGN (t=6.03～14.25, P< 0.01). But there was no significant difference between LN and CGN in SCF and mast cells in renal interstitium. Positive correlation was observed among the expression of SCF and α-SMA and the number of mast cells and macrophages (r=0.47～0.84, P<0.01) at their corresponding locations. The expression of SCF and ot-SMA and the number of macruphages were positively correlated with renal pathological active index, chronic index, albuminuria and the injury of renal interstitium (r=0.34～0.93, P<0.05 or 0.01); meanwhile, it was negatively correlated with Ccr(r=-0.39～0.61, P<0.01). There was significant correlation between SCF, macrophages and anti-dsDNA antibody, complement C3 level, SLE disease activity index (SLEDAI). The number of mast cells in renal interstitium was positively correlated with chronic indexes and the injury of renal interstitium (r=-0.86, r=0.93, P<0.01) and negatively correlated with Ccr (r=-0.56, P<0.01), but not correlated with active index and albuminuria (r=0.27, r=0.23, P>0.05). Conclusion The expression of SCF is widespread in kidney, and it is markedly eorrelated with various kinds of inflammatory cells, renal inherent cells, renal function, and urine protein levels. SCF may be an critical participant in the initiation and progression of renal injuries in human lupus nephritis.

Objective To assess the effects of tacrolimus (FKS06) on podocyte in type 2 diabetic model rats and to explore the potential mechanism.Methods The model rats were fed with high fat and high sugar food and combining with a low-dose of streptozotocin (STZ).They were then randomly divided into a diabetic mellitus group (DM group) and a FK506 group.A normal control group (NC group) was also set.The rats in FK506 group were given with 0.5 mg· kg-1· d-1 FK506 for 8 weeks.The biochemical parameters were measured.The changes of renal pathology and ultrastructure of podocyte were observed by the light and electron microscopy.The expression of nephrin and LC3-Ⅱ was determined by immunohistochemistry and Western blotting.Results (1) Compared with those in NC group,KW/BW,systolic blood pressure (SBP),fasting blood glucose (FBG),triglyceride (TG),total cholesterol (TC),urinary albumin excretion rate (UAE) and creatinine clearance rate (Ccr) in DM group were significantly increased (all P ＜ 0.05).And the KW/BW,UAE and Ccr were decreased in FK506 group compared to those in DM group (all P ＜ 0.05),while other parameters had no significant difference (all P ＞ 0.05).(2) Compared with those in NC group,the glomerular volume,mesangial cell proliferation and accumulation of mesangial matrix were increased,and the foot process became disorder and fusion in DM group,while these changes were significantly reduced in FK506 group.(3) Compared with that in NC group,the expression of nephrin and LC3-Ⅱ was decreased in DM group (all P ＜ 0.05),and both of parameters were higher in FK506 group than those in DM group (all P ＜ 0.05).Conclusion FK506 may enhance podocyte autophagy in type 2 diabetic model rats and attenuate podocyte injury.

Objective To investigate the relationship between interleukin (IL)-18 and podocyte injury of lupus nephritis (LN). Methods Sixty cases of biopsy proven LN patients were enrolled into the study. Thirty cases were selected as controls. The clinical and pathological data, blood and urine samples and renal tissues were collected. The Nephrin expression was detected by immunohistochemical method and IL-18 was measured by enzyme linked immunosorbent assay. The relationship between IL-18 and the Nephrin expression, clinical and pathological indicators of LN were analyzed. Results Thirty-eight cases were in active disease and 22 cases were in inactive disease in LN group according to SLE disease activity index (SLEDAI) 2000. One-way ANOVA showed that the level of plasma and urine IL-18 in the LN groups were higher than those in the control group [(200±38) ng/ml, (18±5) ng/ml] (F=110.84, 203.09, P<0.01). Plasma and urine IL-18 level in active LN group [(565 ±128) ng/ml, (200 ±47) ng/ml] was higher than that in the inactive group [(376 ±106) ng/ml, (67 ±22) ng/ml] (P<0.01), the level of IL-18 of type Ⅳ LN was significantly higher than that of typeⅢand type Ⅴ LN (P<0.01). The expression level of Nephrin in LN groups were lower than those in healthy control group (0.28±0.02)(F=136.39, P<0.01). The expression level of Nephrin in active LN group (0.13±0.03) was lower than that in the inactive group (0.18±0.02) (P<0.01), the level of typeⅣLN Nephrin was significantly 01). The Pearson correlation analysis showed that, compared with plasma IL-18, urine IL-18 level in the LN group was not only negatively correlated with the level of Nephrin and serum C3 (r=-0.780, -0.565, P<0.05), but positively correlated with 24 h UP, erythrocyte sedimentation rate (ESR), SLEDAI and GAI (r=0.546, 0.467, 0.599, 0.634, P<0.05). Serum IL-8 level was independent of albumin (ALB), C4, C reactive protein (CRP) and CI (P>0.05), and was negatively correlated with estimated glomerular filtration rate (eGFR) (r=-0.562, P<0.05). It was positively correlated with serum creatinin, blood urea nitrogen, AI, TLAI and inflammatory cell infiltration (r=0.529, 0.482, 0.665, 0.690, 0.671, P<0.05). Conclusion IL-18 has a very close relationship with podocyte injury in patients with LN, and the uIL-18 can be a potential non-invasive detection method to monitor podocyte injury in LN patients.

ObjectiveTo investigate the protective effect of combination of triptolide and irbesartan on the podocytes in a type 2 diabetic(T2DM) rat model,and evaluate its mechanism.Methods T2DM rats were induced by fed with high-sucrose-high-fat diet combined with a low dose of streptozocin.The rats were randomly divided into 5 groups:normal control group ( NC,n =10),diabetes group ( DM,n =11),triptolide treatment group (DT,n =12),irbesartan treatment group (DI,n =12) and triptolide combined with irbesartan treatment group (DTI,n =13). Ultrastructure of podocytes was observed by electronic microscopy and urinary albumin (UAL) excretion by ELISA was determined after 8 weeks.The expression of nephrin and bone morphogenetic protein-7(BMP-7), connective tissue growth factor (CTGF),transforming growth factor (TGF)β1 mRNA and proteins were detected by immunohistochemistry,real-time PCR and Western blot. Results Increased UAL was significantly attenuated in all treatment groups.Compared to NC group,UAL in DM group was increased significantly (0.45 ± 0.09 vs 6.36 ± 0.87,P ＜ 0.01 ),while decreased in triptolide or irbesartan alone treatment group (2.48 ± 0.37 and 2.68 ±0.42,both P ＜ 0.01 ).Compared with those in control groups,kidney expression of nephrin,BMP-7 mRNA and proteins were downregulated while CTGF, TGFβ1 mRNA and proteins were significantly upregulated in T2DM rats. Triptolide or irbesartan each alone moderately ameliorated albuminuria and podocyte damage.However,their combined usage showed a dramatic therapeutic synergism,manifested by prevention of progressive albuminuria,restoration of the glomerular filtration barrier,reversal of the decline in slit diaphragm proteins,reduction expression of CTGF,TGFβ1,and upregulation of BMP-7.Conclusion Our findings show that triptolide can increase the efficacy of irbesartan,leading to a more effective prevention of kidney disease in T2DM rat model,which may through upregulation of BMP-7 and inhibition the overexpression of CTGF and TGFβ1.