Clinical significance of changes of cardiac troponin T and other biochemical markers in acute ische-mic stroke patients / 中国医师杂志

Objective Through observing the change of serum cTnT level and other biochemical markers such as T3 and TNF-αin acute ischemic stroke patients , to investigate the correlation between the indictors change and short-term prognosis and stroke severity .Methods This study observed 68 patients (male 35 cases ,mean age 68 ±11.4 years; female 33 cases, mean age 69 ±10.3 years).All cases were collected fasting blood sample to test cTnT ,T3 and TNF-α.Death and other unexpected events were recorded in the form of telephone follow-up within one month in survival patients during and discharge from hospital.Results According to cTnT detection level ,all cases were divided into two groups , normal group (54 cases,79.4%) and positive group(14 cases,20.6%).There was statistical significance between posi-tive cTnT and NIHSS score neurological deficits ( p <0.01).cTnT was negratively correlated with T3( r =-0.324 , P <0.05 ) ,but positively with TNF-α( r =0.67 , P =0.017 ) .All patients were followed up for one month.14.7%patients died including 11.7%during hospital and 3% after discharge.After age, NIHSS adjustment cTnT remains an independent risk factor for death ( RR=2.34 ,95%CI=1.22-5.02 , P<0.01 ) .Conclusion After acute ischemic stroke , cTnT is correlated with dropping T 3 level and in-creasing TNF-αlevel, suggesting that both stress and inflammatory response may be involved in heart dam-age.Abnormal elevation of cTnT influenced short-term prognosis of acute ischemic stroke , which can be used as short-term indicators of poor prognosis in the clinical observations .

Construction and expression of the fusional genes Hsp65-Esat6 expressing plasmid of mycobacterium tuberculosis / 中国组织工程研究

BACKGROUND:There is a linker between Hsp65 and Esat6 coding a segment of water repellent polypeptide.It is very gentle and easy to be folded,which is profitable to translate the keno-folding correctly between the two proteins and to make the space structure of fusional proteins consistent with those two native ones.Then,a correct structure is formed and the immunogenicity of the fusional proteins is improved.OBJECTIVE:This study was to clone the fusional genes Hsp65-Esat6 from mycobacterium tuberculosis(MTB)H37Rv, then to construct into a eukaryotic expressing vector which contains an enhanced green fluorescent protein(EGFP) reporter gene,and finally to identify the expression of Hsp65 protein and Esat6 protein by IHC methods.DESIGN:Single sample experiment.SETTING:Department of Microbiology ＆ Immunology,Medical College of Jinan University.MATERIALS:Plasmid pVAE was donated by Professor Cao from South China University of Technology.MTB H37Rv,E.coil DH5 α,expressing plasmid pEGFP-C1 and Hela cells were donated by Doctor Hu Ping.METHODS:The experiment was conducted at the Department of Microbiology ＆ Immunology and the Medical Experimental Center of Jinan University between September 2005 and June 2006.The whole-genome was extracted from MTB H37Rv by molecular cloning technique,and used it as template to amplify Hsp65 (no terminator) gene by polyrnerase chain reaction (PCR),to recombine it with pEGFP-C1 vector after purification to construct pEGHsp65(no terminator)recombination vector.pVAE vector was used as template to amplify Linker-Esat6 gene(with terminator)by PCR, and then to recombine it with pEGHSP65(no terminator)to construct an eukaryotic expressing vector pEGHsp65-Esat6 with the fusional genes Hsp65-Esat6 inside.Finally,genes Hsp65-Esat was checked by molecule biology methods such as PCR, restriction endonuclease and DNA sequencing.Hela cells were transfected with pEGHsp65-Esat6 and the expression of EGFP and the efficiency of transfection were observed.The expressions of Hsp65 protein and Esat6 protein were detected by IHC methods.MAIN OUTCOME MEASURES:①The size of pEGHsp65-Esat6.②The DNA sequencing result of the pEGFP-C1.③The expression of EGFP in the transfected Hela cells.④The IHC results of Hsp65 protein and Esat6 protein in Hela cells.RESULTS:①The size of pEGFP-C1 was 4.7 kb,that of pEGHsp65 was 6.4 kb,and that of pEGHsp65-Esat6 was 6.7 kb.There were differences between their speeds In agarose electrophoresis.②The results showed that it was the same as reported Hsp65 sequence and Esat6 sequence of MTB H37Rv.③After transfecting pEGHsp65-Esat6 for 24 hours,EGFP was found in 30% of Hela through Laser scanning confocal microscope. But there was no EGFP in non-transfected Hela.④Hsp65 protein and Esat6 protein with biological activities were detected in transfected Hela cells by IHC methods.CONCLUSION: Using Hsp65 and Esat6 as immunogens,we have successfully cloned and constructed a eukaryotic expressing vector which contain fusionaI genes Hsp65-Esat6 of MTB H37Rv and EGFP.

Advance in the research on hepatitis C virus non-structural proteins 3 protease inhibitor / 医学研究生学报

In view of the spread of hepatitis C virus(HCV),it still lacks an efficient anti-HCV drug or treatment until now.HCV non-structural proteins 3(NS3) protease is an important drug target in the research of the antiHCV drugs in recent years.This article reviews the active mechanism and peculiarity of HCV NS3 protease inhibitor.

Effect of ATP on proliferation and membrane protein expression in immortalized hmnan fibroblasts / 中国病理生理杂志

AIM: To investigate the effect of ATP on proliferation and membrane protein expression in immortalized human fibroblasts. METHODS: Applying the normal human fibroblast cell line TIG - 7 and OUMS - 36, the immortalized human fibmblast cell line KMST - 6 and SUSM - 1 to culture for 24 hours and 96 hours with the different concentrations of ATP, ADP and AMP, respectively. The number of cell alive, DNA synthesis and the expression of [32p] - ATP labeled proteins in the membrane fraction were also observed. RESULTS: After 96 hours culture, the proliferation inhibition in immortalized cells exposed to 0.4mmol/L ATP was 77 %, while that of normal OUMS36 cells treated with the same concentration of ATP was 41%; the DNA synthesis of the immortalized cells exposed to 0.4mmol/L ATP was greater suppressed than that of normal OUMS - 36 cells. ADP was the only reagent tested that reduced the number of cells significantly( P < 0.01 ). When the immortalized cells exposed to ADP, AMP, adenosine, and phosphoric acid. In the presence of 1 mmol/L ATP, many KMST - 6 cells died, but normal OUMS - 36 cells did not. Compared with normal cells, the expresson of [32p] - ATP labeled 30 kD, 31 kD, 33 kD and 40 kD proteins were markedly higher in the immortalized cells. CONCLUSION: ATP inhibited the proliferation and increased membrane protein expression in immortalized human fibroblasts.

Effect of ATP on proliferation and membrane protein expression in immortalized human fibroblasts / 中国病理生理杂志

AIM: To investigate the effect of ATP on proliferation and membrane protein expression in immortalized human fibroblasts. METHODS: Applying the normal human fibroblast cell line TIG-7 and OUMS-36, the immortalized human fibroblast cell line KMST-6 and SUSM-1 to culture for 24 hours and 96 hours with the different concentrations of ATP,ADP and AMP, respectively. The number of cell alive, DNA synthesis and the expression of [ 32P]-ATP labeled proteins in the membrane fraction were also observed. RESULTS: After 96 hours culture, the proliferation inhibition in immortalized cells exposed to 0.4mmol/L ATP was 77%, while that of normal OUMS-36 cells treated with the same concentration of ATP was 41%; the DNA synthesis of the immortalized cells exposed to 0.4mmol/L ATP was greater suppressed than that of normal OUMS-36 cells. ADP was the only reagent tested that reduced the number of cells significantly(P

Study on p53 gene construction and functional status of PA-1 cells derived from human teratocarcinoma / 中国免疫学杂志

Objective:To understand the construction,functional status and the meaning of p53 gene of PA-1 cells derived from human teratocarcinoma.Methods:DNA basic sequence analysis,FASAY(functional and sequencing analysis of separated alleles in yeast)and Western blot analysis were used.Results:Direct sequence analysis RT-PCR products showed both wild and mutated bands(p53 colon 239 mutation), FASAY showed that one allele of the p53 gene was active(wild) .while the other was inactive( mutant) .In Western blot analysis haven't defect the gene protein of p53, while the PA-1 cells expressed the p21 protein to a lesser extent than normal human tibroblaste. Conclusion:One type of the p53 allele gene of PA-1 cells of human teratocarcinoma was detected as missense mutation after cultured for more than twenty years, and this mutation is not sufficient to induce the instabih'ty of the chromosome of cell line. So that the study for the maintenance contruction of the stability nuclein type of PA-1 cells is very important to determin whether the genes is stability or instability of chromosomes.

Characteristics of chromosome of teratocarcinoma and its influence elements / 中国病理生理杂志

AIM: To investigate the characteristics of chromosome of teratocarcinoma and its influence elements. METHODS: We used the methods of G-banded karyotype analysis, DNA basic sequence analysis and Western blot and the others, and studied the chromosome karyotype and the status of p53 gene of teratocarcinoma PA-1 cell line which had been cultured for 407-445 passages for 20 years. RESULTS: More than 80% PA-1 cells still maintained the near-diploid karyotype ,after passage 30 the cells appeared with M1 and M2 chromosomal markers because of a balanced translocation between chromosomes 15 and 20. DNA directional sequence analysis of RP-PCR products revealed that there were both wild and mutated band (p53 codon 239 mutation), Western blot did not detect mutational p53 gene protein,while p21 protein expression lower than that in normal human fibroblasts. CONCLUSION:Missense mutation of one of the p53 allele gene of PA-1 cells in human teratocarcinoma was detected after cultured for more than twenty years, which was not sufficient to induce the instability of the chromosome of cell line.

Significance and alterations of p14~(ARF) gene of CDKN2A locus in gastric cancer / 中国病理生理杂志

AIM: To investigate the significance and changes of p14 ARF gene in gastric cancer.METHODS: The tumors and gastric tissues neighboring carcinoma from 48 patients with gastric cancer were studied. The homozygous deletions, mutations, methylation of the CpG islands, and mRNA expression of p14 ARF gene were assessed by PCR, PCR-SSCP, PCR based methylation assay, and RT-PCR.RESULTS: ①The homozygous deletion rate of p14 ARF was 31 3% (15/48), and no homozygous deletions were examined in all the gastric tissues neighboring tumor. ②There were no point mutations of p14 ARF in 33 gastric cancers without homozygous deletion and in the matched gastric tissues adjacent to tumor. ③Methylation rate of the CpG islands of p14 ARF was significantly higher in gastric cancers(47.9%, 23/48) than that in gastric tissues neighboring cancer (4.2%, 2/48)( P

Inhibitory effect of ATP on proliferative signaling in immortalized human fibroblasts / 中国病理生理杂志

AIM: To investigate the inhibitory effects of ATP on proliferation signaling in immortalized human fibroblasts. METHODS: Immortalized human fibroblasts were treated with ATP, ATP conbined with calcium or potassium channel antagonists, respectively. The intracelluar-free calcium ([Ca 2+ ]i), inositol 1,4,5-trisphosphate(IP 3) levels and cell viability were detected at different time points. RESULTS: ATP significantly increased the [Ca 2+ ]i and decreased the IP 3 level in immortalized human fibroblasts, especially at initial stage ( P

Experimental study on effects of ATP and its derivatives on the proliferation of the immortalized human fibroblasts / 中国病理生理杂志

MeATP=BzATP, neither 2-MeSATP nor UTP showed any cytotoxicity. No enhanced expression of P 21 was observed and the cell cycle was held in G 1/S in the cells treated with 0.4mmol/L;no cell apoptosis was detected in the cells treated with 1mmol/L ATP. CONCLUSION: Connecting with P 2X or P 2Y purinergic receptors,ATP activated some intracellular signals to inhibit cell growth, the growth inhibition caused by ATP was not due to apoptosis or induction of cyclin/CDK kinase inhibitor,P 21 .

Effect of ATP on the DNA synthesis, membrane protein expression and channel inimmortalized human fibroblasts / 中国免疫学杂志

Abstract Objective:To investigate the effect of ATP on proliferation and membrane protein expression of immortalized human fibro-blasts.Methods: Applying the normal human fibroblast cell line TIG-7 and OUMS-36, the immortalized human fibroblast cell line KMST-6 andSUSM-1 to culture 24 h and 96 h with the different concentration of ATP, ADP and AMP respectively, observed the number of cells alive, DNAsynthesis and the expression of [32 P]-ATP labeled proteins in the membrane fraction. Results: After 96 h cultured, the proliferation inhibition ofimmortalized cells exposed to 0.4 mmol/L ATP was 77% compared with that of the control cultures,while that of normal OUMS-36 cells treatedwith the same concentration was 41%;the DNA sythesis of the immortalized cells exposed to 0.4 mmol/L ATP was greater suppressed thanthat of normal OUMS-36 cells. Treated the immortalized cells with ADP, AMP, adenosine, and phosphoric acid, ADP could reducee the number ofcells significantly(P