The ouabain is a member of the cardiotonic steroids (CT S) that specifically bind to Na+,K+-ATPase. Binding of ouabain to Na+,K +-ATPase causes rapid activation of Src family kinases, and forms a Na+,K +-ATPase-Src complex as a active binary receptor to phosphorylate and assem ble other proteins into different signaling modules.Oubain stimulates hypertroph ic growth in cardiac myocytes and proliferation in smooth muscle cells, it also induces apoptosis and a newly recognized "hybrid cell death" of concurrent apopt osis and necrosisin the same cells.

AIM: To investigate Nrf2 that regulates the expression of ?-glutamylcysteine synthetase(?-GCS) in the inflammatory cells in bronchial asthma guinea pig bronchoalveolar lavage fluid(BALF).METHODS: Adult male guinea pigs were randomly divided into control group(group A),asthmatic group(group B) and dexamethasone group(group C).Asthmatic model was established by the method of ovalbumin challenge.MDA concentration in the lung tissue homogenate was detected.The total cell count and the proportion of inflammatory cells in BALF were measured.The methods of immunohistochemistry and in situ hybridization were used for detection of the protein and mRNA expressions of Nrf2,Bach1 and ?-GCS.RESULTS:(1) The proportion of eosinophils(EOS) in BALF and the MDA concentration of the lung tissue in group B were higher than those in group A and group C.(2) The result of in situ hybridization indicated that the A value of ?-GCS was the highest in group A compared to group B and group C,but the A value of Nrf2 and Bach1 in 3 groups has no statistical significance.(3) Immunohistochemistry indicated that the A value of ?-GCS in group B was lower than that in group A.The positive rate in cell nucleus of Nrf2 in group B was lower than that in group A and group C.The positive rate in cell nucleus of Bach1 in group B was higher than that in group A and group C.(4) The mRNA expression of ?-GCS(A value) showed positive correlation with the positive rate in cell nucleus of Nrf2 and negative correlation with the positive rate in cell nucleus.A negative correlation between the proportion of EOS in BALF of group B and the ?-GCS mRNA was observed.CONCLUSION: There is disequilibrium between oxidation and anti-oxidation in bronchial asthma guinea pig.Inflammatory reaction decreases the expression of ?-GCS in the inflammatory cells in bronchial asthma guinea pig.Dexamethasone regulates the nuclear translocation of Nrf2/Bach1 and increases the expression of ?-GCS.

Whether surfactant protein B (SP-B)-18A/C and 1580C/T polymorphism were associated with susceptibility to chronic obstructive pulmonary disease (COPD) in Chinese Han population was investigated. After genomic DNA was isolated from blood of COPD smokers and control smokers, the genotypes of SP-B-18A/C and SP-B1580C/T polymorphism loci were determined by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) respectively. The results showed that there was significant difference in genotypes distribution frequency of SP-B1580C/T polymorphism locus between COPD smokers and control smokers. C-->T mutation rate (including TT homozygote and CT heterozygote) in COPD smokers was higher than in control smokers (57.9% vs 41.7%, chi2 = 4.93, P<0.05), whereas there was no significant difference in genotypes distribution frequency of SP-B1580-18A/C locus between COPD smokers and control smokers. The allele frequency (29.1%) of SP-B1580-18A/C locus is lower than T allele (70.9%) in Chinese Han Population, and the distribution was different from that in Mexican, in which, the A and T allele frequencies were 85% and 15% respectively. It was concluded that SP-B1580 T allele was probably associated with increased susceptibility to COPD in Chinese Han population; The polymorphism of SP-B-18A/C locus maybe varied with race.
Alleles ; China/ethnology ; Genetic Predisposition to Disease/*genetics ; Genotype ; Polymorphism, Genetic/*genetics ; Pulmonary Disease, Chronic Obstructive/*genetics ; Pulmonary Surfactant-Associated Protein B/*genetics ; Pulmonary Surfactant-Associated Protein B/physiology ; Smoking/genetics

BACKGROUND: Stroke-prone spontaneously hypertensive rat(SHRsp) is an animal model of hypertensive stroke commonly used. The species is hard to be maintained due to hypertension and easily affected by environmental factors, therefore the characteristics of stroke are always aberrated.OBJECTIVE: To observe the abilities of reproduction and maintaining the characteristics of hypertensive stroke in SHPsp.DESIGN: Observational controlled study based on rats.SETTING: Cardiovascular Disease Institute and Fuwai Hospital of Chinese Academy of Medical Sciences.MATERIALS: The experiment was conducted Cardiovascular Disease Institute and Fuwai Hospital of Chinese Academy of Medical Sciences during September 1999 to December 2001. Totally 93 pairs(186) strain SHRsp, 48stroke-prone spontaneously hypertensive rats of 8 weeks age as well as 98pairs(196) rats of normal blood pressure [Wistar Kyoto rat (WKY) ] were selected.METHODS: Brother sister mating method was used. And the conception rate, number of litter size(only calculating the litter size of mother rat not eating neonatal rats), rate of eating neonatal rats by mother rats were recorded and compared with those of WKY. The systolic pressure and heart rate of strain rat were measured when they were 12, 16 and 20 weeks old. In addition, 488-week SHRsp were loaded 10 g/L salty water to accelerate the occurrence of stroke and hypertension and executed when they naturally dead or 12 weeks after salty water load. The brain tissue was processed by H-E staining and observed under microscope to detect the incidence of stroke.MAIN OUTCOME MEASURES: Conception rate of female rats within 2years; litter rate of pregnant rats; eating, rate of eating neonatal rats by mother rats; systolic pressure of strain rat; heart rate; detection rate of stroke.RESULTS: Totally 93 pairs(186) strain rats of SHRsp, 98 pairs(196) WKY and 48 SHRsp of 8 weeks old entered the final analysis. In the first year there were 2 generations delivered, the average conception rate( 100% ) and average litter number (10.3 rats) of SHRsp were higher than those of WKY of the same term(90%, P ＜ 0. 001; 6. 5 rats, P ＜ 0. 001) . In the second year, there were 2 generations of which the conception rate and average litter number(89%, 8.2 rats) of SHRsp were higher than those of WKY(59%, P ＜ 0. 001; 4. 3 rats, P ＜ 0. 001). There were 87 SHRsp female rats and 67normal blood pressure rats pregnant within 2 years, the rate of eating neonatal rats within 4 weeks postnatal was 6% (5/87) which was lower than that of WKY(18%, 12/67), P ＞ 0.05. The systolic pressure of 12weeks old male rats and female rats was 191.6-223.8 mm Hg and 174. 2-196. 3 mm Hg respectively, while that of 16-week old and 20-week male rats were 219.0 -224. 9 mm Hg and 232.0 -242.6 mm Hg respectively. The blood pressure of SHRsp increased with the advancing of age. The heart rate of 12-week old male and female rats was 388-428 times per minute and 373-417times/minute respectively while that of 16-week and 20-week male rats were 392 -410 times per minute and 404-425 times per minute respectively. The pathological detection rate of 48 SHRsp was 81% (39/48).CONCLUSION: The reproduction ability of SHRsp is similar to normal rats. The blood pressure, heart rate and pathological examination of brain tissue of them all maintain the characteristics of hypertension and stroke so that they can be used as qualified experimental model.

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.

To probe into the influence of transplantation of allogenic bone marrow mononuclear cells (BM-MNCs) on the left ventricular remodeling of rat after acute myocardial infarction (AMI), 60 male Wistar rats were evenly divided into three groups at random: control group 1, control group 2 and transplantation group. In control group 1, chest was opened without ligation of coronary artery; in control group 2 and transplantation group, the left anterior descending branch of coronary artery was ligated to establish AMI model. Prepared culture medium and allogenic BM-MNCs suspension were respectively implanted the surrounding area of infracted cardiac muscle via epicardium of control group 2 and transplantation group. Four weeks after the operation, the osteopontin gene (OPN mRNA, P<0.01), type I collagen (P<0.01) and angiotensin II (AngII, P<0.01) content in the left ventricular non-infracted myocardium, and the Ang II density in blood plasma (P<0.05) of transplantation group and control group 2 were all significantly higher than that of control group 1. In the transplantation group, the myocardial OPN mRNA, type I collagen and Ang II content of non-infracted zone in left ventricle, and the Ang II concentration in blood plasma were all significantly lower than those of control group 2 (P<0.05 for all). It is concluded that allogenic BM-MNCs transplantation may ease left ventricular remodeling after AMI by inhibiting the synthesis of type I collagen in the cardiac muscle and down-regulating the expression of Ang II and OPN gene.

Objective To survey the prevalence of chronic obstructive pulmonary disease (COPD)in urban areas of Hunan province and relevant risk factors and provide a basis of the prevention and treatment for COPD. Methods A questionnaire survey was conducted among 4248 residents, aged over 15, by a simple cluster random sampling method in Changsha, Hunan, Wulipai street North Station community. All the respondents filled out an unified epidemiological survey questionnaire. All of the respondents received examination for lung function. Those respondents showed FEV1/FVC ＜70% were further examined by ECG,X ray inspection for differential diagnosis. The data of epidemiological survey was analyzed by multivariate logistic regression method. Results The response rate was 92%. The total prevalence of COPD was 4. 81%.The prevalence of COPD in the males was 6. 6%, and 3. 0% in the females. The prevalence of COPD in the males was significantly higher than that in the females (x2 = 29. 915, P ＜ 0. 01). The prevalence increased with age increasing (P ＜0. 01). The more the education was, the lower the prevalence of COPD was. Risk factors analyzed with non-conditional logistic were as follow. The odd ratio (OR) for COPD in the age was 1.92(P ＜0. 01) and the odd ratio (OR) for COPD in the sex was 1.81 (P ＜0. 01). The weak lighting in house increased the risk with the OR of 4. 25(P ＜0. 01) and pet feeding further increased the risk with the OR of 12.08(P ＜0. 01). The odd ratio (OR) for COPD in the smokers was 1.74(P ＜0. 01) and the prevalence of COPD was related with smoking intensity (branch years of cigarette). Smoking intensity above 500 increased the risk of COPD. The passive smoking increased the risk with the OR of 16. 39(P ＜0. 01). The odd ratio (OR) for COPD in the paternal family history with chronic pulmonary disease was 2. 13(P ＜0. 01) and 2. 11 (P ＜ 0. 01) in the maternal family history. The odd ratio (OR)for COPD in the education degree was 0. 52(P ＜ 0. 01). Conclusions The prevalence of COPD was high in Changsha city, which might be attributed to the risk factors such as house lighting, pet feeding, cooking,aged, male, smoking, passive smoking, and family history. The education degree was the protective factor of COPD. We should intervene the relevant risk factors of COPD so that the prevalence of COPD might be cut down.

Some biological characteristics of human bone marrow mesenchymal stem cells (MSCs) cultured in vitro were observed. hMSCs were isolated from bone marrow and purified by density gradient centrifugation method, and then cultured in vitro. The proliferation and growth characteristics of hMSCs were observed in primary and passage culture. MSCs of passage 3 were examined for the purify by positive rate of CD29 and CD44 through flow cytometry. Human bone marrow MSCs showed active proliferation capacity in vitro. The purify of MSCs separated by our method was higher than 90%. It was concluded that hMSCs have been successfully cultured and expanded effectively. It provided a foundation for further investigation and application of MSCs.
Antigens, CD29/analysis ; Antigens, CD44/analysis ; Bone Marrow Cells/*cytology ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stem Cells/*cytology

OBJECTIVE: The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied. METHOD: The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection. RESULT: The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours. CONCLUSION Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.
Adenoviridae ; Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; Transfection

Objective To promote application and explore the old Chinese tradition of academic experience and their experience side mode.Methods In the study, the model of translational medicine was studied combined with the characteristics of Chinese medicine, taken the model from clinical to basic (experimental) to clinical research, combined with prospective and review of the clinical and research methods, with the modern information technology and other technological, and Tonifying Five Internal Organs(TFIO) byZhang-Chongquanfor treatment of coronary heart disease, vascular dementia.Results The article was Clarified the theory of TFIO byZhang-Chongquan,with clinical evidence.Conclusion The model of translational medicine can be used to summarize the academic experience.