Thioredoxin-interacting protein (TXNIP), also known as vitamin D3-up-regulated protein (VDUP1), is an endogenous inhibitor of thioredoxin (Trx), which regulates the cellular reduction-oxidation (redox) state. TXNIP regulates cellular survival, apoptosis and inflammation induced by glucotoxicity, heat shock and mechanical pressure. The above functions of TXNIP are regulated by carbohydrate response element binding protein (ChREBP) and AMP-dependent protein kinase (AMPK). In recent years, numerous studies showed that TXNIP is involved in diabetes and diabetic complications. On the one hand, TXNIP functions in diabetes by increasing insulin resistance and hepatic gluconeogenesis. TXNIP expression is induced by high glucose, which is implicated in pancreatic beta cell glucotoxicity and endothelial cells dysfunction. TXNIP may contribute to the development and progression of diabetes and its vascular complications. TXNIP may be a new target for diabetes and its vascular complications therapy.
Apoptosis ; Carrier Proteins ; metabolism ; Diabetes Complications ; drug therapy ; metabolism ; Diabetes Mellitus ; drug therapy ; metabolism ; Endothelial Cells ; pathology ; Humans ; Inflammation ; Insulin Resistance ; Insulin-Secreting Cells ; pathology ; Vascular Diseases ; drug therapy ; metabolism

Heart Valve Prosthesis Implantation ; Humans ; Tricuspid Valve Insufficiency ; diagnosis ; etiology ; therapy

Acupuncture Therapy ; instrumentation ; methods ; standards ; China ; Humans ; Moxibustion ; instrumentation ; methods ; standards ; Needles ; standards ; Reference Standards

The baculovirus expression system has been used extensively for the expression of recombinant proteins in insect cells.Recently,recombinant baculoviruses containing mammalian cell-active promoter element have been used to transduce a broad spectrum of primary and established mammalian cells,including non-hepatic cells.Recombinant baculoviruses have been used successfully for transient or stable gene delivery in mammalian cells in vitro,while the efficiency of delivering gene in vivo is inhibited obviously by complements,but efforts have been made to overcome the problems,for instance,VSV-G-pseudotyped baculoviruses display complement resistance.The mechanism of the transduction is still not clearly understood,though many researches have been done.The baculovirus is able to replicate only in insect cells,but it is incapable of initiating replication cycle in mammalian cells,which guarantees high biosafety of this gene delivery system.In addition,this system is easily manipulated and able to carry large inserts.These attributes will undoubtedly lead to the increased application and continued development of this system for highly effective gene delivery into mammalian cells.

To prokaryotic express prokaryotically and to purify the efaA protein from Enterococus faecalis so as to provide the basis for the further study on the pathogenesis and clinical sero-diagnosis of endocarditis caused by E.faecalis, efaA gene of E.faecalis was amplified by PCR, the PCR-amplified product was digested with restriction enzymes and cloned into prokaryotic vector pET32a to construct the recombinant plasmid pET30a/efaA. This recombinant plasmid was confirmed by double enzyme digestion with BamhI and Xhol and then subjected to sequencing, and transformed to E.coli BL21 (DE3). Expression of the fusion protein was induced by IPTG, and analyzed by SDS-PAGE and Western blotting. The recombinant fusion protein was purified by His-binding affinity chromatography.It was shown that efaA gene of 943 bp in size was amplified from Enterococus faecalis and the recombinant plasmid pET30a,/ efaA was successfully constructed and expressed in E.coli BL21. The purified product was found to be 34 kDa in molecular weight as demonstrated by SDS-PAGE and Western blotting. It is evident that the efaA protein of E.faecalis can be successfully expressed and purified.

Objective To investigate the effect of fluvastatin on activation of nuclear factor kappa B (NF-κB)induced by angiotensin Ⅱ (Ang Ⅱ ) in rat kidney tubular epithelial cells (NRK-52E). Methods NRK-52E cells were divided into (1)control group ; (2)Ang Ⅱ groups with different concentration and time; (3)Ang Ⅱ (10-6 mol/L)+SB203580 ( 10 μmol/L)group; (4) Ang Ⅱ (10-6mol/L) +different fluvastatin concentration (10-7, 10-6, 10-5 mol/L)groups;(5)Ang Ⅱ (10-6mol/L) +fluvastatin (10-5 mol/L) +mevalonate (10-4 mol/L)groap. Electrophoretic mobility shift assays (EMSA) was used to detect NF-κB activation. Phosphorylation of cellular p38 mitogen-activated protein kinase (p38MAPK) was determined by Western blot. Monocyte chemoattractant protein (MCP)-1 mRNA was determined by RT-PCR. Results Ang Ⅱ stimulated the DNA-binding activity of NF-κB,phosphorylation of p38MAPK and up-regulated the expression of MCP-1mRNA in cultured NRK-52E cells in a dose-dependent manner (P<0.01). Ang Ⅱ (10-6 mol/L) induced a rapid (5 minutes) elevation of the p38MAPK phosphorylation. NF-κB DNA binding activity was increased at as early as 30 minutes(P<0.01), peaked at 2 hours after AngⅡ treatment (P<0.01). This stimulatory effect of Ang Ⅱ on NF-κB was blocked by SB203580 (a specific inhibitor of p38MAPK) (P<0.01). Incubation of cells with fluvastatin significantly inhibited the Ang Ⅱ-induced NF-κB activation and expression of MCP-1 mRNA in dose-dependent manner (P< 0.05). Exogenous mevalonate (10-4 mol/L) prevented the effect of fluvastatin on NF-κB activation (P <0.05). Conclusions Fluvastatin reduces Ang Ⅱ-induced NF-κB activation via the p38MAPK pathway in NRK-52E cells. Such effect of flurastatin is partly through blocked by mevalonate.

Objective To investigate the effect of xuezhikang on heart in patients with hypertension. Meth-ods 60 systemic hypertension (HT) patients with normal cholesterol were randomized into placebo group and Xu-ezhikang group (1200 mg/d) for 72 weeks. Extended-released nifedipine were administrated by HT patients. Plasma was obtained at baseline, after 24 weeks and 72 weeks of Xuezhikang therapy. Em/Am ratio at atrioventricular ring , E/A ratio at mitral orifice, IVSTd, LVIDd, LVPWTd were measured by echocardiography. Type Ⅰ collagen car-boxypropeptide (PIP) was determined. Results After 72 weeks treatment, the differences of LVWT, IVSTd, LVP-WTd and LVIDd between Xuezhikang group and the control group were not significant; E/A,Em/Am ratios at atrio-ventricoiar ring were significantly increased except that Em/Am at right atrioventricular ring with right ventricular lateral wall(P <0.01 ,P <0.05). After 24 weeks treatment the levels of plasma PIP were significantly decreased in Xuezhikang group compared with the placebo group (P < 0.01, P < 0.05). Conclusion Xuezhikang exerts cooper-ativity in improving those parameters related to myocardial fibrosis so to protect heart function of hypertension pa-tients.

Objective To investigate the role of TLR9 in the glomerulonephritis through observing the changes of Toll-like receptor 9 in the glomerulo nephritis kidney tissue with or without CpG-ODN stimulation .Methods Wistar male rats were randomly divided into normal group(N),nephritis model group(M),CpG-ODN group(CpG) and GpC-ODN group(GpC).The urine samples were collected at 2,4,6 and 8 weeks after treatment,respectively.Blood samples were collected at the end of the last urine sample was collected,and the kidney tissue was collected,then the rats were killed.24 h urine protein was measured by Coomassie light blue technique.Serum album and renal function were determined by serological method.The pathologic changes of kidney were observed by light microscope and NF-?B p65 expression was detected using immunohistochemystry,RT-PCR was performed to detect the expressions of TLR9,INF-? and IL-6 mRNA.Results The expression of TLR9 was lighter in group M,and significantly increased after CpG-ODN stimulation compared with group M.Furthermore,24 h urine protein excretion was markedly increased,serum album was markedly decreased.The histopathologic changes of kidney were more severe.The mesangial cells(MCs) proliferated diffusifully in midrange and wide range,some of the glomeruli formatted cellularity crescent,micrangium loop was limitted,mononuclear macrophile cells were seen in the mesangial region.Conclusion Inflammatory factors mediated by TLR9 can deteriorate the biochemical and histopathologic changes.The immunologic reaction mediated by TLR9 is one of the mechanisms for the glomerulonephtitis' progression.

Objective To observe the effect of fluvastatin on the activity of extracellular signal-regulated kinase(ERK)1/2,expression of connective tissue growth factor(CTGF) in glomerular mesangial cells stimulated by high glucose,and explore the pathogenic mechanism of diabetic nephropathy(DN) and its early prevention and treatment.Methods The study consisted of two parts,the first part was to repeat the existing experienment of stimulatory effect of high glucose on mesangial cells,and validated that high glucose could up-regulated the expression of CTGF through stimulating ERK,and the change was unrelated with hypertonic conditions.There were five groups:①normal control group(NG);②high glucose group(HG);③mannitol group(MN);④NG+ERK inhibitor PD98059 group(NG+PD);⑤HG+ ERK inhibitor PD98059 group(HG+PD).The second part was to observe the changes of the expressions of CTGF protein and mRNA in mesangial cells at high glucose concentration before and after fluvastatin intervention.There were also five groups:①normal control group(NG);②high glucose group(HG);③normal glucose and fluvastatin group(NG+F);④high glucose + fluvastatin(HG+F);⑤high glucose + fluvastatin and mevalonic acid group(HG+F+M).The activity of ERK1/2 and the expressions of CTGF protein and mRNA in cultured glomerular mesangial cells stimulated by high glucose were detected with Western blotting and RT-PCR.The amount of fibronectin(FN) in the supernatant of cells was analyzed by ELISA.Intracellular total protein level was measured by Coomassie brilliant blue method.Results The first part showed that the activity of ERK1/2 and the expression of CTGF protein in HG were more higher than those in NG(P

Objective To explore the role of intercellular adhesion molecule-1(ICAM-1)and lym- phocyte function-associated antigen-I(LFA-1)in the pathogenesis of Sjgren's syndrome(SS)and provide a theoretical basis for clinical therapy.Methods Immunohistochemical method was used to detect these two cellular adhesion molecules in labial salivary glands of primary Sjgren's syndrome patients and 15 healthy controls.Semiquantitative analysis was performed by image analysis software.Results①In salivary gland samples,the expression of both ICAM-1 and LFA-1 was significantly higher compared to that of controls(P