ABSTRACT
AIM:To investigate the possible mechanism of resveratrol(Res)on tumor necrosis factor-α (TNF-α)-induced monocyte chemoattractant protein-1(MCP-1)expression in primary rat pulmonary artery endothelial cells(RPAECs).METHODS: RPAECs were randomly divided into 4 groups: control group, solvent(1% DMSO) group,TNF-αgroup and Res group.Each group was divided into 1 h,4 h and 8 h subgroups(n=6 per time point).The TNF-α+C1142(a rodent chimeric mAb that neutralizes rat MCP-1)group was set up at the 8 h time point.At each time point,the protein and mRNA expression of MCP-1 was measured by Western blot and real-time PCR.RESULTS: Pre-treatment of the RPAECs with C1142 significantly down-regulated the expression of MCP-1(P<0.05).The protein and mRNA expression of MCP-1 was markedly increased in TNF-αgroup(P<0.05).Notably,incubation with Res down-re-gulated the protein and mRNA expression of MCP-1,which was significantly lower than that in TNF-αgroup(P<0.05). CONCLUSION:MCP-1 was involved in the process of TNF-α-induced injury of RPAECs.Res down-regulates the expres-sion of MCP-1 in RPAECs,thus attenuating cell injury.
ABSTRACT
<p><b>OBJECTIVE</b>Lower respiratory tract infections continue to pose a significant threat to human health. It is important to accurately and rapidly detect respiratory bacteria. To compensate for the limits of current respiratory bacteria detection methods, we developed a combination of multiplex polymerase chain reaction (PCR) and capillary electrophoresis (MPCE) assay to detect thirteen bacterial pathogens responsible for lower respiratory tract infections, including Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Mycoplasma pneumoniae, Legionella spp., Bordetella pertussis, Mycobacterium tuberculosis complex, Corynebacterium diphtheriae, and Streptococcus pyogenes.</p><p><b>METHODS</b>Three multiplex PCR reactions were built, and the products were analyzed by capillary electrophoresis using the high-throughput DNA analyzer. The specificity of the MPCE assay was examined and the detection limit was evaluated using DNA samples from each bacterial strain and the simulative samples of each strain. This assay was further evaluated using 152 clinical specimens and compared with real-time PCR reactions. For this assay, three nested-multiplex-PCRs were used to detect these clinical specimens.</p><p><b>RESULTS</b>The detection limits of the MPCE assay for the 13 pathogens were very low and ranged from 10-7 to 10-2 ng/μL. Furthermore, analysis of the 152 clinical specimens yielded a specificity ranging from 96.5%-100.0%, and a sensitivity of 100.0% for the 13 pathogens.</p><p><b>CONCLUSION</b>This study revealed that the MPCE assay is a rapid, reliable, and high-throughput method with high specificity and sensitivity. This assay has great potential in the molecular epidemiological survey of respiratory pathogens.</p>