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Through summarizing and analyzing a large number of documents and reports of large academic conferences in China, the current situation and the prospect of individualized drug delivery model were analyzed based on folate metabolic gene in pharmaceutical care. Folate metabolic genemethylenetetrahydrofolate reductase (MTHFR) C677T and methionine synthase reductase (MTRR) A66G were related with development of multiple diseases. Its polymorphism guidesindividualized drug administrationto increase the efficacy of drugs or decrease adverse effects.
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<p><b>OBJECTIVE</b>To determine expression changes of major outer membrane protein(OMP) antigens of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai during infection of human macrophages and its mechanism.</p><p><b>METHODS</b>OmpR encoding genes and OmpR-related histidine kinase (HK) encoding gene of L.interrogans strain Lai and their functional domains were predicted using bioinformatics technique. mRNA level changes of the leptospiral major OMP-encoding genes before and after infection of human THP-1 macrophages were detected by real-time fluorescence quantitative RT-PCR. Effects of the OmpR-encoding genes and HK-encoding gene on the expression of leptospiral OMPs during infection were determined by HK-peptide antiserum block assay and closantel inhibitive assays.</p><p><b>RESULTS</b>The bioinformatics analysis indicated that LB015 and LB333 were referred to OmpR-encoding genes of the spirochete, while LB014 might act as a OmpR-related HK-encoding gene. After the spirochete infecting THP-1 cells, mRNA levels of leptospiral lipL21, lipL32 and lipL41 genes were rapidly and persistently down-regulated (P <0.01), whereas mRNA levels of leptospiral groEL, mce, loa22 and ligB genes were rapidly but transiently up-regulated (P<0.01). The treatment with closantel and HK-peptide antiserum partly reversed the infection-based down-regulated mRNA levels of lipL21 and lipL48 genes (P <0.01). Moreover, closantel caused a decrease of the infection-based up-regulated mRNA levels of groEL, mce, loa22 and ligB genes (P <0.01).</p><p><b>CONCLUSION</b>Expression levels of L.interrogans strain Lai major OMP antigens present notable changes during infection of human macrophages. There is a group of OmpR-and HK-encoding genes which may play a major role in down-regulation of expression levels of partial OMP antigens during infection.</p>
Assuntos
Humanos , Antígenos de Bactérias , Genética , Metabolismo , Proteínas da Membrana Bacteriana Externa , Genética , Metabolismo , Linhagem Celular , Chaperonina 60 , Genética , Metabolismo , Leptospira interrogans , Genética , Alergia e Imunologia , Virulência , Lipoproteínas , Genética , Metabolismo , Macrófagos , MicrobiologiaRESUMO
In this study, 5'-CMP was sulfonated, and then the modified 5'-CMP was connected to a protein carrier as an immunogen to immunize BALB/c mice. After cell hybridization, screening and recloning , a McAb (B10) with high sensitivity and specificity was selected. In a dot Hot using the McAb B10, less than 0.05 pg of sulfonated DNA could be detected while 10 ng of DNA was not coloured The result showed that the sensitivity of McAb B10 was higher than that of the McAb from "Chemiprobe" kit