AIM:To construct a eukaryotic expression plasmid for enhanced green fluorescent protein(EGFP)and ZNF580 fusion protein,and study its subcellular localization in the transfected MGC803 cells.METHODS:The primers were designed according to the cDNA encoding sequence of ZNF580 full-length open reading frame(1-172aa),ZNF580 amino terminus(1-93aa)and ZNF580 carboxyl terminus(94-172aa).The three cDNA segments of PCR were cloned into pGEM-T vector.Then they were subcloned respectively into plasmid pEGFP-C1(enhanced green fluorescent protein).The subcellular localization of the fusion protein in MGC803 cells transfected with the vector was monitored by autofluorescence microscopy.RESULTS:Restricted enzymes analysis and DNA sequencing showed that the sequences of the pEGFP-ZNF580(1-172),pEGFP-ZNF580(1-93)and pEGFP-ZNF580(94-172)transgenic plasmid were correct.The fusion proteins of EGFP-ZNF580(1-172)and pEGFP-ZNF580(94-172)were localized in the nuclei.CONCLUSION:The recombinant eukaryotic expression vector pEGFP-ZNF580 has been successfully constructed.The nuclear localization signal is within amino acid residues 94 and 172 of ZNF580 carboxyl terminus(C2H2 zinc finger domain).

Objective To investigate the relationship between the genetic polymorphisms of ?_2-adrenergic receptor and the chronic obstructive pulmonary disease.Methods Allele specific-PCR technique was used to determine 16、27 locos of ?_2-AR genetic polymorphism in 64 unrelated patients with chronic obstructive pulmonary disease and 31 healthy controls in the Department of Respiratory Medicine,People's Hospital of Gui zhou province from Mar.2003 to Apr.2004.Results In COPD group and the healthy group the frequency of Gln/Gln genotype of ?_2-AR 27 locus was 42.86%and 19.35%respectively.It showed significantly difference between them(P

Constructing a research oriented learning platform of pathophysiology based on network environment has great significance in cultivating creative medical talents in quality oriented education.This paper aims at the problems of the research oriented learning platform application in students and teachers,and offers some useful advices to be consulted.

Objective To investigate the expression of anti-endothelial cell antibody AECA in the emphysema rats induced by smoking and to analyze the intervention effect of methylprednisolone on it.Methods Thirty-nine rats were randomly divided into the control group,smoking rat emphysema model group (model group) and methylprednisolone intervention group (intervention group).The model group and intervention group conducted the 1-month passive smoking by smog exposure.The intervention group was intraperitoneally injected by methylprednisolone(once daily,6 d per week).After exposing to smog for 90 d,the differences of serum AECA level,IL-8,MMP-9 and TNF-α level in bronchoalveolar lavage fluid(BALF),lung MLI and mean alveolar number (MAN) were compared among groups.Results Compared with the control group and theintervention group,the level of serum AECA in the model group was significantly increased (P＜0.05);Compared with the control group and intervention group,the IL-8,TNF-α and MMP-9 levels of BALF in the model group were also increased(P＜0.05).Conclusion ACEA participates in the smoking induced emphysema formation;methylprednisolone may decrease the level of AECA and cell inflammatory factors and affects the emphysema formation.

Virtual experiments teaching has been characterized by openness, interactivity and re-source sharing. It efficiently improves the experiments teaching effects and promotes the teaching reform. At present the virtual experiment systems used by domestic universities can realize simulation of the ex-perimental principle, apparatus, object, operation and data. In the virtual experiment system students deepen the understanding of the experiments through foregrounding and the networked virtual experiment manage-ment effectively improves the effects of experiments teaching through behind-the-scenes action.

We took measures to construct sound learning mechanism and a set of effective learning system,which were connected tightly with various learning ways according to the idea of ‘ school-based ’ training.Goals of learning mechanism and the set of effective learning system were to improve teacher's learning ability,enhance knowledge and competence of medical college teachers and to construct learning teaching faculty.

Objective To study the association between serum total IgE level and β2-adrenoreceptor polymorphism (β2-AR) in asthmatic patients. Methods ELISA was used to determine serum total IgE and AS-PCR was used to determine β2-adrenoreceptor polymor- phism in 44 asthmatic patients. Results In β2-AR 16 locus genotype, the distribution frequencies of Arg/Arg, Gly/Gly showed increasing tendency, whereas Arg/Gly showed decreasing tendency, in normal serum total IgE group and increased serum total IgE group. But there was no significant difference between this two groups. In β2- AR 27 locus genotype, the distribution frequency of Gln/Gln genotype accounts for 76.2% and Gln/Glu genotype for 19.0% in increased serum total IgE group, while Gln/Gln genotype accounts for 9.0% and Gln/Glu geno- type for 73.9% in normal serum total IgE group. There was significant difference between two groups ( P<0.01 ). Conclusions Serum total IgE was correlated with β2-AR 27 locus genetic polymorphism, which may contribute to understand the mechanism of asthma in the peo- ple of the Han nationality in GuiZhou.

Objective To evaluate the effects of dexmedetomidine (DEX) on cell apoptosis induced by endoplasmic reticulum stress and c-Jun N-terminal kinase (JNK) pathway during one-lung ventilation (OLV) in rats.Methods Sixty male Sprague-Dawley rats were randomly allocated into 6 groups (n =10 each):sham operation group (Sham group),OLV group,OLV + atipamezole (α2 receptor antagonist) group (AD group),OLV + atipamezole + DEX group (DEX+AD group),OLV + low-dose DEX group (DEX-L group) and OLV + high-dose DEX group (DEX-H group).The animals were anesthetized with 10％ chloral hydrate 4.5 ml/kg,tracheostomized and mechanically ventilated.Bilateral lungs were ventilated for 2.5 h in Sham group.The right lung was ventilated for 2.0 h followed by 0.5 h two-lung ventilation in OLV group.In DEX-L and DEX-H groups,DEX was infused intravenously for 1 h at a rate of 2.5 μg · kg-1 · h-1 and 5.0 μg · kg-1 · h-1,respectively,starting from 1 h prior to OLV.Atipamezole 250 μg/kg was injected intravenously at 1 h prior to OLV in AD group.Atipamezole 250 μg/kg was injected intravenously at the onset of DEX infusion (5.0 μg · kg-1 · h-1) in DEX+AD group.The rats were sacrificed and left lungs were removed for determination of weight to dry lung weight ratio (W/D),cell apoptosis in lung tissues (by TUNEL),and expression of glucose-regulated protein 78 (GRP78) mRNA and protein,JNK mRNA and phosphorylated JNK (p-JNK) protein (by RT-PCR and Western blot).Pathological changes of lungs were examined and the injured alveolus rate (IAR) was counted under light microscope.The changes in ultrastructure of lung tissues were observed under transmission electron microscope.Apoptosis index (AI) was calculated.Results W/D,AI and IAR were significantly higher in OLV,AD and DEX+AD group than in Sham group,while lower in DEX-L and DEX-H groups than in OLV,AD and DEX+AD groups.The pathological changes of the structure of lung tissues were observed in OLV,AD and DEX+AD groups,while the pathological changes were significantly alleviated in DEX-L and DEX-H groups.In OLV,AD and DEX + AD groups,there was apoptosis in lots of pulmonary vascular endothelial cells and alveolar epithelial cells,while cell apoptosis was significantly reduced after administration of DEX.The expression of GRP78 mRNA and protein,JNK mRNA and p-JNK protein was significantly higher in OLV,AD and DEX+AD groups than in Sham group,and lower in DEX-L and DEX-H groups than in OLV,AD and DEX +AD groups.Conclusion DEX pretreatment can protect lungs during OLV,and inhibited JNK signaling pathway and reduced cell apoptosis induced by endoplasmic reticulum stress may be involved in the mechanism.

AIM:To elucidate whether ZFP580 is involved in the cardioprotective effects of hypoxic precondi-tioning (HPC) against hypoxia-reoxygenation (H/R) injury in H9c2 myocardial cells.METHODS: Rat heart-derived H9c2 cells were cultured in DMEM .H/R was induced by incubation under ischemic hypoxia for 3 h and reoxygenation for 2 h.HPC was induced by exposing the H 9c2 cells to 10 min of hypoxia and 20 min of reoxygenation for 3 cycles before H/R treatment.MTT staining and LDH leakage detection were used to evaluate the effects of HPC .Western blotting was used to detect the protein levels of ZFP580, phosphorylated ERK1/2 and cleaved caspased-3.The effects of ZFP580 overexpre-ssion or knockdown on H/R induced apoptosis were determined .RESULTS:The results of MTT staining and LDH leakage detection showed evidence of HPC cytoprotection against H /R-induced cell death in H9c2 cells.ZFP580 protein level and ERK1/2 phosphorylation were significantly increased in the HPC group compared with control group and H /R group. PD98059, an inhibitor of ERK1/2 phosphorylation , significantly suppressed the HPC-induced up-regulation of ZFP580 pro-tein expression.ZFP580 overexpression significantly inhibited apoptosis and caspase-3 activation in H9c2 cells.CON-CLUSION:HPC exhibits cytoprotection against H/R and leads to high level of ZFP 580 protein in H9c2 cells.ZFP580 is regulated by ERK1/2 activation and mediates the anti-apoptotic effect of the ERK1/2 signaling pathway in HPC cytoprotec-tion.

Objective To study between serum uric acid and early morning blood pressure of the different degrees of Obstructive sleep apnea syndrome(OSAHS)patients.Methods134 cases of OSAHS were divided into mild,midrange and severe groups according to apnea-hypopnea index(AHI),and 23 healthy people as normal control group.serum uric acid and early morning blood pressure were measured among four groups.ResultsSerum uric acid levels in mild,midrange,severe OSAHS groups and the control group were(392.10?88.22)?mol/L,(460.14?118.86)?mol/L,(537.63?134.11)?mol/L,(304.36?80.12)?mol/L,there were significant differences among four groups.there were significant differences between severe OSAHS and the others in blood pressure,as well as between OSAHS midrange group and the control group.ConclusionSerum uric acid in OSAHS patients increased with increasing degree of AHI,the disorder of serum uric acid possiblly contribute to abnormal blood pressure in OSAHS patients.