Objective]To summarize Professor Tan Zihu’s experience in treating vascular dementia. [Methods]Summarized“the awareness on the etiology and pathogenesis of vascular dementia”,the theory of“prevention of disease”,such as“preventing measure taken before the occurrence of disease”,“treatment characteristics”,and with 1 case for specific instruction. [Result]The responsibility of the etiology’s source of vascular dementia is spleen-kidney deficiency,closely linked to stroke.The basic pathogenesis is the deficiency of sea of marrow,which leads to the dysfunction of brain.He advocates the theory of“prevention of disease”,such as“preventing measure taken before the occurrence of disease”,the primary prevention for the ones who have the high risk factor for stroke,and the early findings,early diagnosis and early prevention of vascular dementia.[Conclusion]Professor Tan Zihu,an expert of TCM,treats vascular dementia starting with spleen-kidney deficiency,combining the theory of“prevention treatment of disease”.It gets good clinical effect with promotion value.

AIM To explore the effect of ACE inhibitor on early kidney hypertrophy and its mechanism in diabetic rats. METHODS Rats were randomly divided into three groups: uninephrectomized rats, streptozotocin induced diabetic rats and diabetic rats treated with benazepril (an ACE inhibitor, 10 mg?kg -1 ?d -1 , ig). Activity of ACE was determined by the fluorimetric assay. Expression of TGF? 1 mRNA and TGF? 1 and p21 CIP1 protein was measured by Northern blot analysis and Western blot analysis, respectively. RESULTS After 1 week, the diabetic rats developed a body weight loss, kidney weight/body weight increased and renal cortex ACE activity elevated despite a decrease in plasma ACE activity. Northern blot analysis showed that renal cortex TGF? 1 mRNA expression in the diabetic rats was enhanced by 1.3 times, compared with uninephrectomized rats. Western blot analysis showed that TGF? 1 and p21 CIP1 protein expression were also increased. Administration of benazepril for one week significantly suppressed kidney hypertrophy. ACE activity in the plasma, renal cortex and medulla was reduced by 89%,70% and 70 5%, respectively. Expression of TGF? 1 mRNA as well as expression of TGF? 1 and p21 CIP1 protein was reduced by 47 7%, 49 5% and 60 0%, respectively. CONCLUSION Our results suggest that the suppression of ACE inhibitor on diabetic kidney hypertrophy might partially be associated with a decrease in the expression of TGF? 1 and p21 CIP1 in diabetic rats renal cortex. However, its exact mechanism remains to be further explored.

Objective To discuss the expression of tumor necrosis factor-like weak inducer of apoptosis ( TWEAK) in serum, urine and renal tissue in lupus nephritis( LN) . Methods The serum and urine levels of TWEAK were assessed by ELISA in LN patients and normal controls. Immunohistochemistry was used to detect the expressions of TWEAK in the kidney of patients and healthy controls. Results The serum and urine concentrations of TWEAK and MCP-1 were significantly higher in LN patients than those in healthy controls(P<0. 01). In addition,the level of serum TWEAK showed positive correlation with SLEDAI and negative correlation with serum C3 level ( P <0. 01),the expression of urine TWEAK showed positive correlation with SLEDAI (P<0. 05). In healthy renal tis-sues TWEAK was distributed mainly in renal tubules and rarely seen in glomerular while the expressions of TWEAK increased in renal tubular and glomeruli stainings were found in LN paitients. TWEAK could induce increased mac-rophage migration, may be involved in the pathological progression of lupus nephritis. Conclusion TWEAK may play key roles in the pathogenesis of LN. Meanwhile, TWEAK in serum and urine may be useful as markers of dis-ease activity. The change of TWEAK maybe associated with the pathology category of LN.

AIM: To study effect of benazepril (an ACE inhibitor) on expression of insulin receptor (IR) and its substrate-1(IRS-1) protein in renal tissue cell membrane in diabetic rats. METHODS: The rats were randomly divided into following groups: control (n=6), streptozotocin induced diabetic (n=7) and diabetic treated with benazepril (n=7). Body weight, kidney weight and kidney weight/body weight were observed after 4 weeks of treatment. ACE activities in plasma, renal tissue were measured by the fluorimetric assay. The expressions of IR and IRS-1 protein were determined by Western blot analysis in renal tissue cell membrane. RESULTS: After 4 weeks of treatment, benazepril significantly ameliorated kidney hypertrophy in diabetic rats. ACE activities in plasma, renal tissue were redunced by approximately 92.00% and 88.77%, respectively. Western blot analysis showed that the expressions of IR and IRS-1 protein were increased by 2.1 and 1.5 folds in renal tissue cell membrane in diabetic rats. However, benazepril reduced expression of IR and IRS-1 protein by 45.74% and 47.66%, respectively. CONCLUSIONS: Increased expression of IR and IRS-1 protein might be related to abnormally active glucose metabolism in diabetic rat kidney. Down-regulation of expression of IR and IRS-1 protein might be one of important machnisms of Benazepril nephroprotection on diabetic rats.

Objective　To evaluate the efficiency of low molecular weight heparin(LMWH, fraxiparine) given as a single predialysis bolus injection with reused dialyzers in comparison with standard heparin(SH) administered with a continuous infusion. Methods　30 hemodialysis patients were studied in a radomized crossover fashion. Dialyzers fibrer bundle volumes(FBV), predialysis hemocrit and 2 h clearance of urea, creatinine were observed in the first, the fourth dialysis. The plasma heparin activities(anti-Fxa levels) were measured by the chromogenic substrate assay in 0 h, 2 h, 4 h of dialysis. Results　Significant increase (P<0.05) was seen in the number of dialyzer reuse in LMWH group compared with SH group; there was not significant decrease in dialysis FBV as well as 2 h clearance of urea, creatinine (P>0.05); in addition, the plasma heparin activity(anti-Fxa levels) were comparable in both groups after 2 h of dialysis, however, they were significantly higher after 4 h in the LMWH than those in the SH group (P<0.05). Conclusion　LMWH as a single bolus dose can prevent decrease in dialyzer clearance. It is clinically worthy of further popularity.

AIM To study effects of ACE inhibitor on the ex pression of GluT4 mRNA and protein in diabetic rat heart. METHODS The following groups of rats were randomly designed: control rats, diabetic ra ts and diabetic rats treated with benazepril (an ACE inhibitor). After 4 weeks o f treatment, ACE activities were determined by fluorimetric assay in plasma and heart; Myocardial GluT4 mRNA expression were measured by Northern blot analysis; GluT4 protein expression were measured by Western blot analysis. RESULT S After 4 weeks of treatment, there was a significant increase in myocard ial ACE activities despite a trend toward reduce in plasma activitics, ACE activ ities were inhibited by 92 1%, 89 0% in plasma and heart, respectively; Northe rn blot analysis revealed that the expression of GluT4 mRNA was reduced in diabe tic rat heart by 40%, and treatment with benazepril did not prevent the diabetes -induced reduction of GluT4 mRNA; However, Western blot analysis revealed that benazepril did prevent the diabetes-induced reduction of GluT4 protein in cell membrane of diabetic rat heart. CONCLUSION Benazepril could significantly suppress ACE activities in diabetic rat heart, bu t it did not influence the expression of the myocardial GluT4 mRNA. However, it did prevent the diabetes-induced reduction of GluT4 protein in cell membrane of rat heart.

Objective To investigate the expression of transforming growth factor ?_1(TGF?_1)in renal cortex from uninephrectomized diabetic rats. Methods Wistar rats were divided into uninephrectomized rats(group A), streptozotocin diabetic rat(group B). Blood glucose, serum insulin level and body weight, kidney weight, kidney weight/body weight as well as renal tissue protein contents were observed after 1, 4 weeks of streptozotocin injection. The expression of TGF?_1, precollagen 1?(Ⅳ) and fibronectin mRNA were measured by Northern blot analysis, and TGF?_1 protein by Western blot analysis in kidney cortex. In addition, ACE activities were determined by fluorimetric assay in plasma, kidney cortex and medulla. Results Group B demonstrated significantly elevated blood glucose and decreased serum insulin level. Kidney weight、kidney weight/body weight and renal tissue protein contents progressively increased despite total body weight loss. There was significant(P

Aim To investigate the effect of paeoniflorin(PF)on TLR2/4 pathway in AGEs-induced RAW264.7 macrophages.Methods RAW264.7 macrophages were incubated at different time points in AGEs stimulation,as well as different concentrations of PF,to optimize experimental conditions.RAW264.7 macrophages were randomly divided into five groups: control group(DMEM),bull serum albumin(BSA)group(200 mg·L-1 BSA),AGEs group(200 mg·L-1 AGEs),paeoniflorin group(200 mg·L-1 AGEs+10-5 mol·L-1 PF)and TLR2/4 inhibitor group(200 mg·L-1 AGEs+30 mg·L-1 OxPAPC).The expression of Toll-like receptor 2(TLR2),Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),p-IRAK1,TIR-domain containing adaptor protein-inducing IFN-β(TRIF),interferon regulatory factor 3(IRF3),p-IRF3,NF-κB p-p65,NF-κB p65,inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α),interleukin-l β(IL-1β)and monocyte chemotactic protein-1(MCP-1)were measured by Western blot.Real-time PCR was used to detect the expression of TLR2 and TLR4 mRNA,while TNF-α,IL-1β and MCP-1 levels in cell supernatant were measured by ELISA.Results Compared with control group,AGEs significantly increased the expression of TLR2,TLR4,MyD88,p-IRAK1,TRIF,IRF3,p-IRF3,NF-κB p-p65,NF-κB p65,iNOS,TNF-α,IL-1β and MCP-1 proteins(P<0.01),as well as TLR2 and TLR4 mRNA(P<0.01).TNF-α,IL-1β and MCP-1 contents were also elevated in cell supernatant(P<0.01).The effects induced by AGEs were decreased significantly in PF and TLR2/4 inhibitor group(P<0.01).Conclusion PF plays an anti-inflammatory effect via inhibiting TLR2/4 pathway on macrophages,which may provide a new theoretical basis for the treatment of diabetic nephropathy.

Objective To investigate the regulation of melatonin (MT) on Toll-like receptor 4 (TLR4) signaling in diabetic db/db mice kidneys.Methods The 48 10-week-old male db/db mice were randomly divided into db/db group,db/db+MT 50 μg/kg group,db/db+MT 100 μg/kg group and db/db+MT 200 μg/kg group,each consisting of 12 mice.These mice received i.p.injections of MT These mice received i.p.injections of MT [dissoved in phosphate buffer solution (PBS)/ dimethylsulfoxide (DMSO) solution,given every day].Alternatively,12 db/m mice served as the control group.db/m and db/db group were injected i.p.with the same volume of PBS/DMSO solution.The animals were sacrificed after 12 weeks of dosage administration.Blood glucose (BG),body weight (BW),kidney weight (KW) and 24 h urinary albumin excretion rate (UAER) were determined;Kidney pathological lesions were evaluated by renal pathological staining.Immunohistochemistry of renal TLR4,NF-κB p65,and ED-1 was performed to determine the immunoreactivity.Western blotting was used to detect the expression of renal TLR4,myeloid differentiation factor 88 (MyD88),TIR-domaincontaining adaptor inducing interferon-β (TRIF),interferon regulatory factor 3 (IRF-3) and NF-κB p65,while the mRNA expressions of renal tumor necrosis factor-α (TNF-α) and monocyte chemotactic protein-1 (MCP-1) were evaluated by real-time PCR.Results Compared with control group,the levels of BG,BW,KW and UAER were much higher in db/db mice group (P ＜ 0.01),while KW in db/db+MT (100,200 μg/kg) groups and UAER level in db/db+MT (50,100,200 μg/kg) groups were distinctly decreased compared with those in db/db group (P ＜ 0.01).In week 12 db/db mice,the glomerular mesangial expansion index and tubulointerstitial injury index were increased compared with those in db/m mice (P ＜ 0.01).The above kidney histopathologic lesions were distinctly ameliorated by 50,100,200 μg/kg MT (P ＜ 0.05).Immunohistochemistry intensity of renal TLR4,NF-κB p65 and ED-1 displayed obvious differences between db/m mice and db/db mice (P ＜ 0.01),and that were remarkably decreased in db/db+MT (50,100,200 μg/kg) mice compared with db/db mice (P ＜ 0.05).Western blotting showed that the protein expression of renal TLR4,MyD88,TRIF,IRF-3 and NF-κB p65 were stronger in db/db group compared with those in db/m group (P ＜ 0.05) and weaker in db/db+ MT (50,100,200 μg/kg) groups compared with those in db/db group (P ＜ 0.05).Futhermore,the mRNA expressions of renal MCP-1 and TNF-α were higher in db/db group compared with those in db/m group (P ＜ 0.01) and lower in db/db+MT (50,100,200 μg/kg) groups compared with those in db/db group (P ＜ 0.01).Conclusion Melatonin may partly down-regulate TLR4 signaling pathway to inhibit Inflammatory reaction and alleviate kidney injury in diabetic db/db mice.

ObjectiveTo investigate the effect of tacrolimus (FK506) on macrophage accumulation,proliferation and activation in the kidney of early diabetic rats and to explore its possible mechanism of renal protection.Methods Rats were randomly divided into control,model and tacrolimus groups.Diabetic model rats were induced with intraperitoneal injection of streptozotocin.Tacrolimus(0.5 or 1.0 mg·kg-1 ·d-1) was orally administered once a day for 4 weeks.Kidney weight index(KWI),24-h urinary albumin excretion rate(UAER) and creatinine clearance rate(Ccr) were measured.Kidney pathology was observed by light microscopy.ED-1,PCNAandiNOSpositivemacrophagesweredetectedbysingleanddoublestainingof immunohistochemistry.Results KWI increased in model group and was significantly reduced by tacrolimus treatment with 1.0 mg·kg-1 ·d-1 (P＜0.05).UAER elevated in model group and was markedly attenuated by tacrolimus treatment with 0.5 and 1.0 mg·kg-1 ·d-1 (P＜0.05).Elevated glomerular volume of model rats was significantly decreased by tacrolimus treatment with 0.5 and 1.0 mg·kg-1·d-1 (P＜0.05),and increased indices of tubulointerstitial injury were only ameliorated by 1.0 mg·kg-1·d-1 tacrolimus(P＜0.01).Marked accumulation of ED-1+ cells in diabetic kidney was found,which was not inhibited by tacrolimus treatment with 0.5 and 1.0 mg·kg-1·d-1.ED-1PCNA+ cells and ED-1+ iNOS+ cells were significantly elevated in kidneys of model group,while they were significantly inhibited by tacrohmus treatment with 0.5 and 1.0 mg·kg-1·d-1 (P＜0.01).Conclusion Tacrolimus can ameliorate early renal injury of diabetic rats and its mechanism may be partly associated with the suppression of increased macrophages activation.