[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 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 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 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.

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.

HBx gene is a multifunctional regulator,which has extensive trans-activating effects,and can activate transcription factors,inhibit DNA repair and regulate cell proliferation,differentiation and apoptosis.In recent years,the role of HBx gene in pathogenesis of hepatitis B virus-associated glomemlonephritis (HBV-GN) has been extensively studied,and the results show that HBx can promote glomerular mesangial cell proliferation,and induce damage or apoptosis of podocytes and renal tubular epithelial cells.This paper reviews the research progress on biological characteristics of HBx and its role in pathogenesis of HBV-GN.

Objective To analyze the relationship between the expression of protection of telomeres 1 (POT1) and the pathogenesis of acute myeloid leukemia (AML). Methods 62 patients with de novo AML (case group) and 10 patients with iron deficiency anemia (control group) were enrolled in this study. The quantitative real-time polymerase chain reaction (PCR) and Western blot were used to detect the expression of POT1 in AML patients. Results There were 62 de novo AML patients, including 2 cases M1, 14 cases M2, 12 cases M3, 14 cases M4, 17 cases M5, 2 cases M6 and 1 case AML without classification. According to the risk stratification, high risk group (24 cases), medium risk group (22 cases) and low risk group (16 cases) were divided. Compared with that in the controls, POT1 expression levels in patients with AML were significantly decreased both in mRNA and protein level (P< 0.05). The relative expression levels of POT1 mRNA and protein in patients with M2, M4 and M5 were significantly lower than those in the controls (P< 0.05). The expression levels of POT1 in high risk group, medium risk group and low risk group were significantly decreased than those in the controls (P<0.05). Compared with that in the controls, The relative POT1 mRNA expression was significantly decreased in M3 patients (P< 0.05), but not in protein level. POT1 protein expression was showed both in the cytoplasm and nucleus. There was no significant difference of the expression of POT1 protein between cytoplasm and nucleus (P> 0.05). Conclusions POT1 may be involved in the pathogenesis of AML. POT1 protein expresses in both cytoplasm and nucleus, and the regulatory mechanism may be related to the telomere length.

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.

BACKGROUND:Transient receptor potential channel C6 (TRPC6) is a new and important slit diaphragm-associated protein in podocytes involved in regulating glomerular filter function. Glomerular TRPC6 expression is closely associated with proteinuria in diabetic kidney disease. OBJECTIVE: To investigate the expression of canonical TRPC6 in mouse podocytes induced by high glucose, and to explore the possible mechanism of diabetic kidney disease. METHODS:Mouse podocyte cels were cultured and divided into normal glucose group (5.6 mmol/L D-glucose), normal control group (5.6 mmol/L D-glucose+25 mmol/L mannitol) and experimental groups which were in the environment of high glucose (30 mmol/L). The experimental groups included high glucose group, valsartan treatment groups (10-5 mol/L) and U73122 control group (10μmol/L U73122). After 48 hours, the expressions of mRNA and proteins of TRPC6, nephrin and angiotensin II (AngII) were detected respectively by real-time quantitative PCR and western blot analysis. RESULTS AND CONCLUSION:Compared with the normal control group, the expressions of mRNA and proteins of TRPC6 and angiotensin II were markedly elevated in the high glucose group (P < 0.01), while the expressions of mRNA and proteins of nephrin were decreased (P < 0.01). The mRNA and proteins of TRPC6 and angiotensin II expressions were significantly down-regulated by valsartan (P < 0.05,P < 0.01), while the mRNA and protein expressions of nephrin were effectively up-regulated (P < 0.05). Compared with the high glucose group, the expressions of mRNA and proteins of TRPC6 and angiotensin II were ameliorated in the U73122 control group. The expressions of mRNA and proteins of TRPC6, nephrin and angiotensin II had no statistical significance between the normal control group and normal glucose group (P > 0.05). Angiotensin II-TRPC6 signaling pathway may mediate high glucose-induced podocyte injury, meanwhile it provides a new theoretical basis for the treatment of diabetic kidney disease, by which the angiotensin receptor blockers can protect podocytes in diabetic kidney disease.