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.

Objective: To get a general estimation of the pneumatic system in ventilators and supply technical assistance for screening and removing the faults of pneumatic component. Methods: Pneumatic system of three kinds of ventilators, Sevro-i/s, Rapheal XTC and PB840 were analyzed and the differences of the three pneumatic systems were compared. Results:Known the construct and differences of pneumatic system. Conclusion: Pneumatic system is the major component of ventilator, and the analysis of the pneumatic component is benefited for the common troubleshooting of ventilators.

Objective　To evaluate the effects of vitrectomy associated with extraction of intraocular foreign bodies in the posterior segment.Methods　Retrospective analysis was done on clinical records of 11 patients (11 eyes) having undergone vitrectomy company with extraction of intraocular foreign bodies in the posterior segment.Results　The intraocular foreign bodies in the posterior segment were extracted successfully in all cases, the retinal detachment and endophthalmitis were cured. Postoperation visual acuity was improved in 8 eyes and did not change in 3 eyes. None suffered from postoperative secondary intraocular hemorrahge, decompensation of corneal endothelium and other serious complications.Conclusion　Vitrectomy combined with extraction of intraocular foreign bodies in the posterior segment is less damaging, accurate and safe, and enhances the cure rate. It is an effective method for treating intraocular foreign bodies.

Objective To develop techniques based on polymerase chain reaction(PCR) which can detect 2 of most common deletional ? thalassemia ? 3.7 deletion and ? 4.2 deletion in China accurately and speedily. Methods Two groups of primers were designed and synthesized. PCR conditions were optimized. The PCR pro ducts were analysed by 1.0% agarose gel electrophoresis. The gel was stained by EB and photographed using an UVP gel documentation. Results PCR product of a 1 700 bp DNA fragment with primers A′, B′, C3 indicates the ? 3.7 deletion while a 1 900 bp fragment indicates a normal or wild type of ? globin gene. Occurrence of the 1 700 bp and 1 900 bp simultaneously indicates a heterozygous of the ? 3.7 deletion. Neither of the 2 bands was presented, indicating a homozygous of South East Asia type of deletion (-? SEA ). According to patterns of 1 580 bp and 1 180 bp hand amplified by a PCR with primer G′, E, F′, we detected the ? 4.2 deletion and distinquished its heterozygous and homozygous. Conclusions The 2 PCR based techniques developed in our laboratory are accurate, simple, well reproducible and easy to use for screening of the 2 deletion types of ? thalassemia determinants.

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 study the changes in heme oxygenase-1/endogenous carbon monoxide system in rabbit aorta with atherosclerosis induced by high cholesterol diet, and the influence of the system on the atherosclerotic process. Methods Rabbits were given 1.5% cholesterol diet (Ch group, n=8) or 1.5% cholesterol diet plus hemin(15mg?kg~ -1 ?d~ -1 , ip., Hm group, n=8) or zincprotoporphyrin IX (Znpp-IX,45?mol?kg~ -1 ?d~ -1 , ip., Zn group, n=8).After 12 weeks,serum lipids and total bilirubin, aortic CO protduction and HO activity were assessed, the area of atherosclerotic plaques in aorta were measured by oil-red-O staining and image analysis software. Results Compared with control group (C group, n=8), aortic HO activity and CO production were higher significantly, and the aortic plaques area was (54.00?4.16)% in Ch group. Znpp-IX injection markedly decreased aortic HO-1 activity and CO production, the aortic plaques area was (61.13?3.50)%. Compared with Ch group, hemin injection increased markedly not only serum total bilirubin concentration, but also aortic HO activity and CO production, and the aortic plaques area was (17.88?3.01)%. Conclusion The HO-1/CO system may play a protective role against atherosclerosis. Enhancement of HO-1 activity is possibly the basic function of HO-1/CO system in prevention of atherosclerosis.

Objective:To investigate the feature of depression in patients with Parkinson's disease(PD) and the related factors.Methods:Use Hamilton Depression Scale(HAMD) to measure the level of depression of 60 patients with PD.Functional state of the patients was evaluated by Motor Dysfunction Rating Scale for Parkinson's Disease(MDRSPD).Results:29 cases(48.3%) were identified to have depression,among whom 40% were of minor or moderate degree,8.3% were severe.The main manifestations of depression were anxiety/body,retardation,hopelessness and insomnia.Scores of hopelessness,retardation,and insomnia factor were significantly higher in the rigidity and tremor combining rigidity patients than those in tremor patients.The level of depression positively correlated with the duration of disease and degree of dysfunction due to PD,and it has no relation with age,sex,and education.Conclusions:The incidence of depression was higher in patients with PD.PD patients of different clinical types had different characteristics of depression.The rigidity type is more likely to get depression.The degrees are closely related to duration of disease and degree of dysfunetion.

Objective To study the effects of puerarin on advanced glycation end products (AGEs) and expression of monocyte chemoattractant protein-1 (MCP-1) in diabetic rats. Methods The rats were randomly divided into normal control rats (CON rats), diabetic rats (DM rats), diabetic rats treated with aminoguanidine (AG rats), and diabetic rats treated with puerarin (PU rats). The diabetic rat model was induced by streptozotocin. The serum levels of AGEs and MCP-1 were quantified by fluorescence spectroscopy and ELISA respectively. Tissue sections with PAS staining and electronic microscopy were used for observation of the pathologic changes in renal tissues. Immunohistochemistry was applied to detect the expression of MCP-1 protein in renal cortex. Results Serum levels of AGEs and MCP-1 in DM rats were higher than those in PU rats and AG rats (both P

The crowd were divided to 3 groups according to glucose tolerance:normal glucose tolerance,impaired glucose regulation,and new type 2 diabetes,the data was analyzed.The result showed that as the severity of abnormal glucose metabolism,the serum GGT gradually advanced,early insulin secretion index descended,The levels of resistance index,fasting and postprandial blood sugar,and glycosylated hemoglobin all raised.The serum GGT and early insulin secretion index was inversely correlated.The higher GGT level was an independent risk factors of abnormal glucose metabolism.

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.