ABSTRACT
<p><b>OBJECTIVE</b>To investigate the mechanisms for the cytopathic effect (CPE) of human cytomegalovirus (HCMV) in ECV304 endothelial-like cells.</p><p><b>METHODS</b>PCR and indirect immunofluorescence were used to detect HCMV infection by examining immediate-early (IE) gene and protein expression of the virus in ECV304 cells. Phase-contrast and electron microscopies were performed to observe the morphological changes of the infected and uninfected cells, and DNA ladder analysis and flow cytometry were carried out to study HCMV-induced cell apoptosis.</p><p><b>RESULTS</b>In HCMV-infected ECV304 cells, cytopathic effects were first observed at approximately 72 h post-infection. The cells with CPE changes exhibited detachment from the monolayer, cell rounding and shrinkage. The expression of the IE gene was detected. Chromatin condensation and nuclear fragmentation along with dramatic changes of the mitochondria were observed by electron microscopy at 96 h post-infection. Cellular DNA fragmentation was observed in the infected cells, which had cells apoptotic rates of 4.1% and 45.7% at 96 h and 144 h post-infection, respectively.</p><p><b>CONCLUSION</b>HCMV can induce apoptosis of ECV304 endothelial-like cells.</p>
Subject(s)
Humans , Antigens, Viral , Genetics , Metabolism , Apoptosis , Physiology , Cell Line , Cytomegalovirus , Genetics , Metabolism , DNA, Viral , Endothelial Cells , Cell Biology , Virology , Fluorescent Antibody Technique, Indirect , Immediate-Early Proteins , Genetics , Metabolism , Microscopy, Electron , Microscopy, Phase-Contrast , Polymerase Chain Reaction , Umbilical Veins , Cell Biology , VirologyABSTRACT
<p><b>OBJECTIVE</b>To observe the pathological changes and morphological alterations of ECV304 cells after the infection by herpes simplex virus type 2 (HSV-2) in vitro.</p><p><b>METHODS</b>Passaged ECV304 cells were infected with HSV-2, TCID50 and morphological changes were observed by optical microscopy and tissue staining.</p><p><b>RESULTS</b>One day after HSV-2 infection, swelling, rounding, and increase of thickened cytoplasmic granules occurred in the ECV304 cells, and on day 2, cell fusion was observed with weakened nuclear staining.</p><p><b>CONCLUSION</b>ECV304 cells mostly undergo necrosis after HSV-2 infection without obvious evidence of cell apoptosis.</p>
Subject(s)
Humans , Cells, Cultured , Endothelium, Vascular , Pathology , Virology , Herpesviridae Infections , Pathology , Herpesvirus 2, Human , Necrosis , Umbilical Veins , PathologyABSTRACT
The effects of two different sample labeling methods on background signal intensities for high-density 60mer oligonucleotide microarray were investigated. Peripheral blood samples from five disease and five control subjects were collected. Total RNA targets from peripheral blood mononuclear cells were extracted and labeled with RD-PCR protocol, which were hybridized to Agilent Human 1B oligonucleotide microarrays in a two-color comparative format. The positive control targets were labeled with the directly incorporated fluorescently-labeled dNTP labeling. The SPSS program was performed to test normality of the dataset, variance homogeneity between the groups, coefficients of variation (CV) and analysis of variance. The results showed that the background signal intensities of Cy3 channel were higher than those of Cy5 channel. The differences of background signal intensities between the RD-PCR approach and the directly incorporated fluorescently-labeled dNTP labeling were extremely significant (P- Cy3