Objective To explore the effects of high glucose on glucose transport activity, and phosphorylationandexpressionofinsulinsignalingproteins in primary cultured rat adipocytes. Methods Isolated rat adipocytes were cultured at different glucose concentrations (5, 10, 15, 25 mmol/L) for 24 h. Then the glucose uptake, the phosphorylations of insulin receptor (IR), insulin receptor substrate (IRS) 1 and 2 and protein kinase B (PKB) as well as the protein expressions of IRS1, IRS2, p85 subunit of phosphatitylinositol 3 kinase (p85) and PKB were measured. Results These adipocytes treated with different high glucose showed the impairment of the basal and insulin induced increase in glucose uptake and significant decrease of IR, IRS1 and PKB phosphorylations as well as IRS1 protein expression, but up regulation of IRS2 protein expression. p85 and PKB contents and IRS2 phosphorylation were unaffected. Conclusion The exposure to high glucose inhibits glucose uptake and induces insulin resistance in adipocyte. The mechanism may be involved in affecting the multiple step phosphorylations and the expressions of insulin signaling proteins.

Type 1 diabetes is pathologically characterized by autoimmune insulitis-related islet β-cell destruction.Although intensive insulin therapy for patients with type 1 diabetes can correct hyperglycemia,this therapy does not effectively prevent diabetes-related cardiovascular complications.Hence,preservation of natural β-cell function is critical for the prevention of diabetes-related complications.Receut studies have shown that autologous hematopoietic stem cell transplantation ( A HSCT) is a new promising approach for the treatment of type 1 diabetes by reeonstitution of immunotolerance and preservation of islet β-cell function.Here we introduce the key results of a clinical trial ( NCT 01341899,ClinicalTrials.gov) performed in Chinese type 1 diabetes patients ( most of them with diabetic ketoacidosis at onset ) and discuss the potential mechanisms underlying the action of HSCT and its prospects in the clinical management of type 1 diabetes.

Objective To assess the effect of heat shock protein 70 of BCG (BCG HSP70) gene transfection on tumorigenicity and immunogenicity of murine lymphocytic leukemia cells (L1210). Methods BCG HSP70 gene was transfected onto the surface of murine lymphocytic leukemia cells (L1210) by lipofectamine 2000. And then the positive clone (L1210-HSP70) highly expressing HSP70 was selected as the tumor vaccine to study the tumorigenicity experiments in nude mice and syngeneic mice, the therapeutic experiments, and the immunoprotective effects. Results The expression of BCG HSP70 on the L1210 cells surface was detected, and the L1210-HSP70 cells had the same tumorigenicity as the parental L1210 cells did. Tumorigenicity experiments in syngeneic mice:In L1210-HSP70 group, tumor growth was slow or without the formation of tumor. As compared with L1210 group and L1210-neo group the mice survival time was signiifcantly prolonged, showing a marked stimulating effect on L1210 specific Th1 cells,. Tumor-bearing mice showed complete coagulation necrosis and abundant CD8+T lymphocyte inifltration (P<0.05). The tumor vaccine of L1210-HSP70 cells had the antitumor therapeutic efifcacy and immune protection effect, demonstrating that the tumor growth was signiifcantly inhibited, tumor diameter was markedly reduced and the survival time of tumor-bearing DBA/2 mice was further prolonged. Conclusions BCG HSP70 gene transfection could effectively improve the immunogenicity of tumor cells, activate speciifc T cells and enhance the anti-tumor immunity in vivo. Meanwhile, the host anti-tumor immunity could be enhanced.

Objective To explore the value of heart-type fatty acid-binding protein (H-FABP) in the early diagnosis of acute myocardial infarction (AMI). Methods Sixty confirmed AMI patients were observed, the data of H-FABP, cardiac troponin T (cTnT), creatine kinase MB (CK-MB) were detected in < 6 h and 6-12 h after the symptoms appeared, and the sensitivity of the three markers was calculated. The specificity was compared with 15 uncertain AMI patients and 45 healthy subjects. Results The early diagnosis sensitivity of H-FABP in < 6 h was 94% in AMI, which was higher than that of cTnT (50%) and CK-MB (56%) (P < 0.05 ). The diagnosis sensitivity of H-FABP, CTnT, CK-MB in 6-12 h was 100%, 92%, 92% respectively (P> 0.05 ). There was no significant difference in the specificity among the three markers (P > 0.05). Conclusions H-FABP has more sensitivity and specificity in the early diagnosis of AMI. It is applicable in the screening of patients who suffered chest pain and the diagnosis of early AMI.

A total of 1 007 inpatients with type 2 diabetes mellitus (T2DM) were divided into 2 groups,according to whether there was accompanied with nonalcoholic fatty liver(NAFLD) ; and the data of the two groups were analysed.Serum uric acid levels were positively correlated with body mass index and blood urea nitrogen and creatinine,while negatively correlated with HbA1c.T2DM patients complicated with NAFLD were younger,had shorter duration of T2DM,and showed impaired uric acid clearance with raised serum uric acid.The prevalence of NAFLD was higher with increasing serum uric acid levels.

Objectives To prepare the HL-60 cell vaccine expressing heat shock protein 70 (HSP70) of Bacille calmette-Guérin (BCG), so as to study its anti-tumor effect and mechanism. Methods The whole BCG HSP70 gene was amplified from BCG genome by polymerase chain reaction (PCR) and sub-cloned into the polyclone endonuclease sites in pDisplay. The recom-binant vector of pDisplay-HSP70 was verified by sequencing. Then the HL-60 cell vaccine expressing the protein onto the cell surface was prepared by lipofectamine transfection. To detect the immunogenicity of HL-60 cells expressing HSP70, the test groups were divided into three subgroups, HL60-wt, HL60-pDisplay, and HL60-HSP70 respectively. Each group was cultured with peripheral blood T cells for 72 h, then the proliferation indices of T cells were assayed by CFSE-staining method, and IFN-γwere tested by enzyme-linked immunosorbent assay (ELISA). The HL-60 cells of different groups were cultured with peripher-al blood T cells for 6d. The wild-type HL-60 cells were added and co-cultured for another 12h. Cytotoxicity assay was measured by LDH release. Results (1) The fragment of BCG HSP70 was consistent with the theoretical value. DNA sequencing showed that the recombinant vector of pDisplay-HSP70 was correctly constructed. (2) BCG HSP70 expressed onto the HL-60 cells sur-face. (3) Detection of the immunogenicity: ①The most significant T cell proliferation was observed in the group of HSP70-transfected HL-60 cells (P<0.05). There was no difference between the HL60-wt group and HL60-pDisplay group (P>0.05).②The contents of IFN-γof the HSP70-HL60 group was the highest.③The inhibiting activity of CTLs on HL-60 cells in the group of HSP70-transfected HL-60 cells was more significant than that of wide-type and pDisplay--transfected HL-60 cells. And with the increase of the E:T ratio, the inhibiting activity of CTLs in the HSP70-HL60 group was rising. Conclusions The recombinant eukaryotic expression vector (pDisplay-HSP70) of BCG HSP70 was successfully constructed. And the HL-60 cell vaccine expressing BCG HSP70 onto its surface was successfully prepared. The results showed that gene transfection of BCG HSP70 could significantly enhance the immunogenicity of HL-60 cells.

Objective To explore the correlation between BODE index with fatigue symptoms on COPD patients,and provide new ways for clinical evaluation,prediction,symptom control and establishment of effective management mode.Methods 120 COPD patients of stable stage were selected to be investigated and analyzed by the Fatigue Scale-14(FS-14),six-minute walking distance(6MWD),pulmonary func-tion test and the body mass index (BMI).Dyspnea was measured using the Medical Research Council (MRC) dyspnea scale.Results The fatigue symptoms showed high positive linear correlation with BODE index and mMRC,and significant negative correlation with 6 MWD,FEV1％ and BMI.Conclusions The study shows that fatigue symptoms had a higher prevalence in COPD patients of stable stage.There were a high correlation between fatigue symptoms with the BODE index,mMRC,6MWD,FEV1％ and BMI.The BODE index was a good predictor and evaluation of fatigue symptoms.

Objective:To establish the quality standards for Zhichuan adhesive plasters;Methods:Herba ephedrae、Bergenin arolisiae、Flos caryophylli、Semen sinapis in Zhichuan adhesive plasters were identified by TLC. Ephedrine hydrachloride in Herba ephedrea was determined by RP HPLC.Results:These methods were simple and accurate.Conclusion: The methods can be used for the quality control of Zhichuan adhesive plasters.

Objective To observe the effect of advanced glycosylation end products(AGEs) on gene expression of connective tissue growth factor(CTGF) in NIH Swiss mouse embryo fibroblasts(NIH/3T3),and to assess the intervention actions of aminoguanidine(AG) and puerarin(Pue) on CTGF mRNA expression in NIH/3T3.Methods AGEs were synthesized by coincubation of BSA with glucose.The AGEs content was measured by fluorescence spectroscopy.NIH/3T3 cells were treated with AGEs(prepared with 20,50,80 mmol?L-1 glucose) for 24 h.The NIH/3T3 cells were treated with AGEs(prepared with 50 mmol?L-1 glucose) for 0,6,12,24 and 48 h.The intervention actions of AG and Pue with different concentration(0.25,0.5,1.0 and 1.5 g?L-1) were evaluated.The CTGF mRNA expression in NIH/3T3 was determined by RT-PCR.Results Compared with BSA control,the CTGF mRNA expression levels in NIH/3T3 were increased by treatment with AGEs(prepared with 20,50,80 mmol?L-1 glucose) for 24 h(P

Objective To observe the changes of left ventricular diastole function in the old patients with paroxysmal atrial fibrillation(PAF)and probe the value of multiindexes to evaluate the left ventricular diastole function.Methods Us the ultrasonic color doppler to determinate the different indexes of left ventricular diastole function in the thirty-four older patients with PAF after converting who came to my hospital from September 2004 to February 2006,and the healthy elder were the controls.Results The index of isovolumic relaxation time(IVRT),flowing velocity E wave(EV),flowing velocity A wave(AV)and the ratio of EV and AV(EV/AV)of the group with PAF after converting were higher significantly in PAF group after converting(99?11 ms,75?13 ms),(0.65?0.14 m/s,0.84?0.15 m/s),(0.77?0.12 m/s,0.55?0.13 m/s)and(0.84?0.13〈1,1.52?0.16〉1)than those in control group.(P〈0.01).Conclusion A significant failure of left ventricular diastole function exists in the old patients with PAF and the multiindexes to evaluate the left ventricular diastole function are more objective.