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BACKGROUND:Human amniotic epithelial cells have some properties of stem cells, which can be induced to differentiate into corneal epithelial cells, but cannot be tracedin vitro. OBJECTIVE:To investigate the feasibility and infection efficiency of adenovirus vector carrying enhanced green fluorescent protein (EGFP) into the human amniotic epithelial cells. METHODS:The adenovirus vectors carrying EGFP was transferred into human amniotic epithelial cells culturedin vitro. After cultured and amplified, the morphology difference between transfected and non-transfected human amniotic epithelial cels was observed. The transfected human amniotic epithelial cells were observed under fluorescence microscope, and the cell cycle and the expression rate of EGFP in transfected human amniotic epithelial cells were detected by flow cytometry. RESULTS AND CONCLUSION:No obvious difference in the cell morphology was found between transfected human amniotic epithelial cells and normal human amniotic epithelial cells cultured in vitro. Flow cytometry analysis revealed that the EGFP positive rate was highest and reached up to 99.01% at 48 hours after transient transfection. The cell cycle of human amniotic epithelial cells transfected by the adenovirus vector was slowed a bit. To conclude, the adenovirus vector is a good medium of transfecting EGFP into human amniotic epithelial cells, and makes it more convenient to observe the further transformation of human amniotic epithelial celsin vitro.
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Objective:To study the current status and needs of Community Care for disabled elderly in Beijing and put forward the suggestions for improving the elderly Community Care system. Methods:Descriptive statistical a-nalysis, Chi-square test and Multivariate logistic analysis were adopted to analyze the situation of Community Care for Disabled elderly using data from the community survey of elderly population in Beijing. Results:The status of health is not optimistic for Disabled elderly, there are high demands for the community health services and welfare facilities for elderly and the needs for services provided by part-time home care, day care and volunteers are high, but those services and facilities provided by communities were found to be inadequate. Conclusion:China has a large popula-tion base of Disabled elderly and its number grows fast, but community care supply is in shortage and cannot meet the needs of those people. It is therefore urgent to improve community health services and community care system.
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AIM:To investigate the effects of resveratrol ( Res) on the proliferation of ARPE-19 cells and to ex-plore the possible mechanisms.METHODS:After ARPE-19 cells were treated with Res at concentrations of 0, 50, 100, 150, 200 and 300 μmol/L for 24 h, 48 h and 72 h, the effects of Res on the proliferation of the cells were tested by CCK-8 assay.The ARPE-19 cells were treated with Res at concentrations of 0, 100, 150 and 200 μmol/L for 48 h.The effects of Res on the cell cycle and apoptosis were detected by flow cytometry with Annexin V-FITC/PI staining.The protein expression of proliferating cell nuclear antigen (PCNA) was detected by immunofluorescent assay.The mRNA expression of PCNA, P21 and P27 was determined by real-time PCR.RESULTS:The results of CCK-8 assay showed that Res inhibited the prolifera-tion of ARPE-19 cells in a time-and dose-dependent manner.The treatment with Res for 48 h resulted in an arrest of cell cycle at S phase without increasing cell apoptosis.Res inhibited the protein expression of PCNA in ARPE-19 cells.The re-sults of real-time PCR showed that Res increased the mRNA expression of P21 and P27, and decreased the mRNA expres-sion of PCNA.CONCLUSION: Res inhibits the proliferation of ARPE-19 cells and induces the cell cycle arrest at S phase.The mechanism may be related to up-regulation of P21 and P27, and down-regulation of PCNA.
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[ ABSTRACT] AIM: To determine the aberrant methylation status in the gene promoter regions of CDH13, RASSF1A, DLEC1, SEPT9 and RUNX3 by detecting the plasma specimens and the value of their combined detection for di-agnosis of lung cancers.METHODS:Nest methylation specific PCR ( nMSP) was used to detect the promoter methylation status of the 5 genes in the plasma from 106 normal controls, lung cancer tissues, lung benign tissues and the plasma from 106 patients with lung cancers.Multiple displacement amplification ( MDA) was used to amplify modified genomic DNA to solve the problem of insufficient of plasma DNA template.RESULTS: The positive rates of promoter methylation of CDH13, RASSF1A, DLEC1, SEPT9 and RUNX3 in the lung cancer tissues were 51.9%, 44.3%, 54.7%, 36.8%, 24.5%, respectively, and those in the plasma were 46.2%, 41.5%, 50.9%, 31.1%, 19.8%, respectively.The re-sults of the Kappa consistency check showed that the lung cancer tissues and the plasma had obviously coherence in the methylation status of the 5 gene promoter regions.Combination of DLEC1, CDH13, RASSF1A, and SEPT9 had a higher di-agnostic efficiency than the others, as their ACC value was 0.8208 and youden index was 0.6415 ( with the sensitivity of 81.13% and the specificity of 83.02%) .CONCLUSION:Combination detection of promoter methylation of lung cancer-related genes in the plasma is expected to apply to the early diagnosis of lung cancer.