ABSTRACT
<p><b>OBJECTIVE</b>Genetic evolution of VP1 of enterovirus type 71 in Shenzhen were analyzed.</p><p><b>METHODS</b>All samples were tested by RT-PCR using EV71 specific primer. The VP1 of EV71 were amplified and sequenced. A phylogenetic tree was constructed by comparison of the sequences with subgenotype A, B and C using DNAStar, BioEdit and Mega 3.1 software.</p><p><b>RESULTS</b>Among 35 strains, the homogeneity of the VP1 nucleotide sequence was between 92.1%-100%. The homogeneity of the VP1 nucleotide sequence with subgenotype A and B was between 81.4% -91.1%. The VP1 nucleotide sequence of 35 strains of Shenzhen shared between 93% -97.4% identity with cluster C4. The prevalence strains of EV71 were cluster C4b from 1998 to 2004, and gradually moved to C4a since 2003. All of EV71 were C4b from 2006 to 2008. Also, the homogeneity of the VP1 nucleotide sequence with Anhui FY23 EV71 strain were 94.5% -94.7%, 95.7% -95.8%, 96.2%, 95.4% -97.5%, 96.3% -99.2% from 2003 to 2008. It shows that the homogeneity was increased year by year. There was a mutation (A --> C) at No. 66 nucleotide of VP1 of EV71 that two strains were isolated in 2003 and 8 strains in 2008, that caused amino acid mutation (Q --> H) at No. 22 of VP1.</p><p><b>CONCLUSION</b>EV71 C4b was gradually moved to C4a from 1998 to 2008. There was a missense mutation at No. 66 nucleotide of VP1.</p>
Subject(s)
Humans , Enterovirus , Classification , Genetics , Mutation, Missense , Genetics , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Viral Structural Proteins , Classification , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the binding profile of NV strain SZ9711 (GII-4) with human histo-blood group antigens (HBGAs).</p><p><b>METHODS</b>The P domain-encoding fragment was amplified by RT-PCR from the stain SZ9711 and cloned into the pGEX-4T-1 vector. The recombinant fusion protein was expressed in E. coli and purified using the column Sepharose 4B. The P protein was released by thrombin cleavage. The binding of P particles of SZ9711 and VA387 with the HBGAs were measured by saliva-based EIA method.</p><p><b>RESULTS</b>The expression of the recombinant fusion protein was shown by the SDS-PAGE, in which a 38 x 10(3)-P protein was obtained. Saliva-based EIA revealed that the P particle of SZ9711 bound to HBGAs in saliva similar to that of the strain VA387 reported previously. It bound strongly to saliva of type A, B and O(secretor) but did not interact with saliva of type O(non-secretor). Noteworthy, binding ability of SZ9711 P particle to type A saliva was lower than that of the VA387 P particle.</p><p><b>CONCLUSION</b>This is the first time that a P particle was prepared from a norovirus strain isolated in China and the binding ability of the P particle with HBGAs was analyzed. The result indicated the binding profile of the SZ9711 P particle was similar to that of VA387 reported previously. These data may be valuable in studying the relationship between noroviruses and their bindings to HGBA receptors.</p>
Subject(s)
Humans , Blood Group Antigens , Metabolism , Caliciviridae Infections , Metabolism , Virology , China , Norovirus , Chemistry , Genetics , Metabolism , Protein Binding , Saliva , Chemistry , Metabolism , Viral Proteins , Genetics , MetabolismABSTRACT
Genetic characteristics of enterovirus type 71 in Shenzhen from 2005 to 2008 were analyzed. All samples were detected by RT-PCR using EV71-specific primers. The VP1s of EV71 strains were amplified and sequenced. A phylogenetic tree was constructed by comparison of the sequences with those of subgenotype A, B and C using DNASTAR, BioEdit and Mega 3.1 softwares. The VP1 nucleotide sequences of 17 strains of Shenzhen shared 95.3%-99.4% identities with cluster C4a, and one strain shared 93.0%-95.6% identities with cluster C4b. The homogeneity of the VP1 nucleotide sequence with subgenotype A and B was 81.8%-86.0%. Among 18 strains, the homogeneity of the VP1 nucleotide sequence was between 92.5%-100%. All EV71 strains circulating in Shenzhen were of subgenotype C4. The predominant strain of EV71 belonged to cluster C4a, also there was cluster C4b.
Subject(s)
Humans , Cell Line , China , Enterovirus A, Human , Classification , Genetics , Enterovirus Infections , Virology , Feces , Virology , Genotype , Molecular Sequence Data , Phylogeny , Viral Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the chemical constituents of Mentha spicata.</p><p><b>METHOD</b>The chemical constituents were isolated by silica gel column chromatography, and identified by physical and chemical characters and spectroscopic analysis.</p><p><b>RESULT</b>Compounds I - V were obtained and their structures were elucidated as protocatechuic aldehyde (I), protocatechuic acid (II), chrysoeriol (III), 5, 6-dihydroxy-7, 8, 3', 4'-tetramethoxyflavone (IV), nodifloretin (V).</p><p><b>CONCLUSION</b>Compound I and II were first isolated from the genus Mentha. Compound Ill, IV and V were isolated from M. spicata for the first time.</p>