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 culture the acute leukemia cells in vitro,and to prepare cancer vaccine expressing heat shock protein 70 (HSP70) of Bacille Calemette-Geérin(BCG) onto the cell surface,so as to study its anti-tumor effect and mechanism.Methods Acute myeloid leukemia (AML) cells were cultured in a serum-free Stemspan(H) culture supplemented with cytokines [stem cell factor(SCF),flt-3 ligand (FL),interleukin (IL)-3 and IL-6] in vitro.And B-lineage acute lymphoblastic leukemia (B-ALL) cells were cultured in a Iscove modified medium(IMDM) culture supplemented with cytokines (SCF,FL,IL-3 and IL-7) in vitro.Cellular morphology was observed by the microscopy and immunophenotype determination was used to verify the biological characteristics of acute leukemia cells after culture.Lipofectamine 2000 was used to transfect the pDisplay-HSP70 plasmid into acute leukemia cells.The expression of HSP70 on the cell surface was detected by fluorescene microscope.Then the immunogenicity of the leukemia cells expressing HSP70 were detected.The experimental groups were divided into 3 subgroups:the wide-type acute leukemia cells (wt-LC group),the pDisplay-leukemia cells (pDisplay-LC group),and the pDisplay-HSP70-leukemia cells (HSP70-LC group),respectively.The leukemia cells in different groups were cultured with autologous peripheral blood T cells for 72 hours.The proliferation indices of T cells were assayed by carboxyfluorescein diacetate succinimidyl ester (CFSE)-staining method,and the contents of interferon-γ(IFN-γ) were tested by enzyme-linked immunosorbent assay (ELISA).The leukemia cells in different groups were cultured with autologous peripheral blood T cells,and after 6 days,the fresh acute leukemia cells were added [in the different ratios of cytotoxicity T lymphocyte (CTL):leukemia cells were 10 ∶ 1,20 ∶ 1,40 ∶ 1 and 80 ∶ 1] and continued to be cultured for another 12 hours.Cytotoxicity assay was measured by lactate dehydrogenase (LDH) release.Results After short term culture in vitro,the leukemia cells were in colony-like suspension and maintain the proliferation characteristics were maintained.The cell proliferation was rapidly cultured for about 10 days and then was gradually slowed down.But there was no difference between the day 10 and day 0 in the expressions of CD13 and CD33 in fifteen cases of AML cells (P ＞ 0.05).Equally,there was no difference between the day 10 and day 0 in the expressions of CD19,CD10 and CD22 in fifteen cases of B-ALL cells (P ＞ 0.05).After BCG HSP70 gene transfection,the yellow-green fluorescence on the leukemia cells surface was observed under the confocal microscope.Detection of the immunogenicity:(1) Autologous T cell proliferation:the most significant T cell proliferation was observed in the group of HSP70-transfected leukemia cells (t =17.89,19.58,all P ＜0.05).There was no difference between the wt-LC group and pDisplay-LC group (P ＞ 0.05).(2) The contents of cytokines:the IFN-γ level in the group of HSP70-transfected leukemia cells was higher than those of wide-type acute leukemia cells and the pDisplay-transfected ones (t =24.72,24.81,all P ＜ 0.05).(3) Cytotoicity of CTL:the killing rate in HSP70-transfected leukemia cells was significantly higher than those of wide-type acute leukemia cells and pDisplay——transfected ones(F =13.66,P ＜ 0.05).And with the increase of the ratio from 10 ∶ 1 to 80 ∶ 1,the inhibiting activity of CTL in the HSP70-LC group was raising(F =19.69,P ＜ 0.05).Conclusions Fresh acute leukemia cells can be successfully cultured in vitro.Short-term culture can significantly increase the number of leukemia cells,but has little effect on surface antigen expression.So,the biological characteristics of the leukemia cells can be maintained.The leukemia cells vaccine expressing BCG HSP70 onto its surface was successfully prepared,and gene transfection of BCG HSP70 can significantly enhance the immunogenicity of leukemia cells.

Objective To explore the clinical value of examining HPV E6/E7 mRNA level in assessing cervical le?sions infected with high-risk human papillomavirus (HR-HPV). Methods The cervical epithelial cells were collected from 265 patients with HR-HPV infection, including 100 cases of neoplasia free/inflammation group (control group), 88 cas?es of cervical intraepithelial neoplasia (CIN)Ⅰ, 33 cases of CINⅡ, 28 cases of CINⅢand 16 cases of cervical carcinoma and the transcription of HPV E6/E7 mRNA level was examined using branched DNA (b-DNA) technology. Results The positive rate HPV E6/E7 mRNA were higher in CIN Ⅱ(81.82%), CINⅢ(89.29%) and cervical cancer group (100.00%) than tthat in control group (20.00%) and CINⅠ(35.23%) with significant difference, and there were no significant differences between other groups;The positive rate and transcription level of HPV E6/E7 mRNA in HSIL (high grade squamous intraepi?thelial lesion)and cancer group were significantly higher than normal, ASC(atypical squamous cell carcinoma) and LSIL(low grade squamous intraepithelial lesion) group (P<0.05). Conclusion The transcription level of HPV E6/E7 mRNA may re?flect the activity of the virus and the progression of disease, and could be use as an effective indicator to screen high grade cervical pathological changes and a complementary method of cervical lesion screening.

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.

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.

0.05).②The animal experiment showed ,in contrast to the control , embryonate rats’ rhodocyte SOD were significantly different after 2 days but no difference before prenatal. ③in contrast with the control , SOD of placenta were significantly different only after 9 days. Conclusions Diagnostic ultrasound in early pregnancy can make SOD change ,but the changes can be reparable.

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 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