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

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 .

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

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.

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.

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.

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.

Percutaneous pulmonary valve implantation (PPVI) is an exciting growing field in cardiovascular medicine.With the sophisticated improvement of biomedical engineering,development of PPVI offers people a new way of less invasive techniques to treat these groups of patients with pulmonary valvular regurgitation over the past several years.New stent designs and operative and interventional hybrid approaches are under investigation.This nonsurgical approach has been proven to be feasible and holdpromise,although many obstacles still exist.At the present time,PPVI has been widely used in clinical in the United States and European countries,but in our country,only a small number of cases reports have been published.With wider application and development of PPVI,cumulative experiences and long-term follow-up of PPVI therapies,better criteria will be established for patient election.Complications will be reduced and safer and more effective treatment results will be achieved.PPVI therapies will be applied to a larger subset of patients and become an important alternative to future conventional pulmonary valve replacement.

Objective To investigate the application value of low kilovolt technique combined with lower contrast in the head and neck angiography by the second-generation dual source CT.Methods From October 2013 to January 2015,120 patients undergoing head and neck computed tomography angiography (CTA) were randomly divided into groups A,B,C,D.Each group was given a different tube current and different dose of contrast agent and saline solution 50 ml.Group A received current 100 mA and contrast agent 50 ml,group B received current 100 mA and contrast agent 40 ml,group C received current 80 mA and contrast agent 50 ml,and group D received current 80 mA and contrast agent 40 ml.CT values of aortic arch,bilateral common carotid artery bifurcation,double sided M1 segment of middle cerebral artery,basilar artery enhanced degree of the straight sinus were measured in each group.Residual artifacts caused by contrast agent in brachiocephalic vein and subclavian vein were observed.5-score method was used to assess the quality of reconstructed image,and the radiation exposure dose was calculated.Results The mean effective dose was reduced by 29％ in group C as compared with group A [(0.53±0.17) mGy vs.(0.74±0.04) mGy].There was no significant difference in developing strength in cerebral arteries angiography between group C and group A (P=0.247),but the inage noise was slightly larger in group C than group A,without significant differences (P=0.203).The average effective dose in group A was almost the same as that in group B [(0.74 ± 0.04) mGy vs.(0.73 ± 0.05) mGy].Structure display of cerebral arteries on CT volume rendering (VR) and multiplane reformation (MPR) images had no significant differences between group A and group B (P=0.114).The average effective dose in group C was almost the same as that in group D[(0.53 ± 0.17) mGy vs.(0.53 ±0.01) mGy].Structure display of cerebral arteries on CT volume rendering (VR) and muhiplane reformation (MPR) images had no significant differences between group C and group D (P=0.109).The mean effective dose was reduced by 28％ in group D as compared with group B[(0.53±0.01)mGy vs.(0.73±0.05) mGy].There was no significant difference in developing strength in cerebral arteries angiography between group C and group A (P=0.236),but the image noise was sligbtly larger in group C than group A (P =0.212).Conclusions Application of low tube current combined with low concentration of contrast agent in the head and neck dual-source CT angiography is feasible in clinical diagnosis,with good clinical value.It can not only get better image quality,but also meet the needs of clinical diagnosis.

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