ABSTRACT
To study the impact of the enterovirus 71(EV71) on the nuclear transport mechanism,The pGFP-NLS vector with nuclear location signal(NLS) was constructed, RD cells transfected by the pGFP-NLS vector were inoculated with the EV71 or cotransfected by EV71-2A vector. The results showed that GFP protein with NLS was expressed in the cytoplasm due to the inhibition of nuclear transport. In order to further study the mechanism of the EV71 to prevent nuclear transport,Nup62 was detected by Western blotting after RD cells were infected with EV71 or transfected by EV71-2A vector. The results showed that decreased expression of Nup62 could be detected after infection with EV71 and transfection by EV71-2A vector. This study demonstrates that the cleavage of Nup62 by EV71 2A protease may be the mechanism of nuclear transport inhibition.
Subject(s)
Humans , Active Transport, Cell Nucleus , Cell Line, Tumor , Cell Nucleus , Metabolism , Enterovirus A, Human , Genetics , Metabolism , Enterovirus Infections , Virology , Gene Expression Regulation, Viral , Genetic Vectors , Green Fluorescent Proteins , Metabolism , Membrane Glycoproteins , Metabolism , Nuclear Localization Signals , Metabolism , Nuclear Pore Complex Proteins , Metabolism , Peptide Hydrolases , Metabolism , Recombinant Fusion Proteins , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To study the isoflavonoid constituents of the roots of Astragalus membranaceus var. mongholicus.</p><p><b>METHOD</b>Such column chromatography methods as HPD-100 macroporous adsorption resin, silica gel, polyamide and Sephadex LH-20 gel were used for seperating and purifying isoflavonoids, and their structures were identified on the basis of spectral data.</p><p><b>RESULT</b>Fourteen compounds were separated and identified as: formononetin (1), ononin (2) calycosin (3), calycosin-7-O-beta-3-D-glucopyranoside (4), (6aR, 11aR)-3-hydroxy-9,10-dimethoxypterocarpan (5), (6aR, 11aR)-3-hydroxy-9,10-dimethoxypterocarpan-3-O-beta-D-glucopyranoside (6), (3R) -7,2'-dihydroxy-3', 4'-dimethoxyisoflavan (7), (3R) -7, 2'-dihydroxy-3', 4'-dimethoxyisoflavan-7-O-beta-D-glucopyranoside (8), 6"-O-acetyl-ononin (9), 6"-O-acetyl-(3R) -7, 2'-dihydroxy-3', 4'-dimethoxyisoflavan-7-O-beta-D-glucopyranoside (10), 6"-O-acetyl-(6aR, 11aR)-3-hydroxy-9, 10-dimethoxypterocarpan-3-O-beta-D-glucopyranoside (11), pratensein (12), sissotrin (13) and 5,7,4'-trihydroxy-3'-methoxyisoflavone (14).</p><p><b>CONCLUSION</b>Compound 10 was a new compound. Compounds 9, 11, 13,14 were separated from A. membranaceus var. mongholicus for the first time.</p>