RESUMEN
To explore the relationship between the genetic diversity and biological functional site of human immunodeficiency virus HIV-1 gp120 and the pathogenesis of AIDS dementia complex (ADC), the full length sequences of gp120 gene was amplified with PCR from genomic DNA which was extracted from lymphoid and different brain department (periaortic lymphoid, temporal gray/white matter junction, periventricular tissue, choroids plexus, occipital white matter and occipital gray/white matter junction.) of a patient who died of ADC. PCR products were cloned into the pGEM-T vector and positive clones were sequenced. The analysis of neighbor-joining tree, N-glycosylation sites, values of ds/dn, and loop were then all performed. The samples were all identified as HIV-1 B and genetic variation existed in HIV-1 gp120 isolated from different tissues. Compared with standard HIV-1B gp120, biological functional sites of HIV-1 gp120 isolated from the patient changed to some extent. In addition, there were differences in some biological functional sites of HIV-1 gp120 between lymphoid and brain. Therefore, genetic diversity and alterations of some biological functional sites of HIV-1 gp120 might be associated with the pathogenesis of ADC.
Asunto(s)
Humanos , Complejo SIDA Demencia , Virología , Secuencia de Aminoácidos , Variación Genética , Genética , Proteína gp120 de Envoltorio del VIH , Química , Clasificación , Genética , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Homología de Secuencia de AminoácidoRESUMEN
<p><b>OBJECTIVE</b>To construct the cloning vector of glycoprotein G2 gene of hantavirus (HV), to analyze the sequence of G2 gene by the phylogenetic tree, and to study the differences among glycoprotein G2 genes from the world around.</p><p><b>METHODS</b>Envelope glycoprotein G2 gene was amplified from four specimens of Shandong province by RT-PCR, and the product recombined into the PMD-18T vector. The clones that carry the G2 gene were identified. After sequencing, the gene sequence was handled with the software DNASTAR, compared with 24 strains worldwide and the phylogenetic tree was drawn.</p><p><b>RESULTS</b>HV G2 gene was amplified by RT-PCR from 4 specimens, named GM04-38.G2, ZB8.G2, JUN5-14.G2, RCH5.G2, respectively. The map of the phylogenetic tree showed that all the 4 strains belonged to SEO-type hantavirus. The analysis of the sequence showed that all the four HV strains had the highest rates of homology with Z37 strain. The sequence homology of SEO-type HV strains was from 82.3% to 99.8%.</p><p><b>CONCLUSION</b>The four cloning vectors containing the glycoprotein G2 genes were successfully constructed. Envelope glycoprotein G2 gene of four specimens from Shandong province had high homology rates.</p>
Asunto(s)
Animales , Ratones , China , Clonación Molecular , Orthohantavirus , Genética , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Proteínas del Envoltorio Viral , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To know the anti-viral effects of rhubarb ethanol extract (REE) on herpes simplex virus(HSV) infection in vivo.</p><p><b>METHODS</b>BALB/c mice inoculated from tail vein with 0.15 ml of HSV (TCID50=10(3)) were injected hypodermically with REE next day. After divided into seven groups, three groups of mice were given different doses of REE respectively and the other groups as controls. Pathological sections from the liver, spleen, kidney were made at different times of postinfection, and their pathological changes were observed under microscope; the virus titers in viscera were assayed by using plaque formation technique and the rhubarb inhibitions to the infection of HSV in vivo?were observed.</p><p><b>RESULTS</b>No toxic response to mice were observed for REE injected hypodermically; no pathological changes were observed in different therapy groups of spleens. And those in livers and kidneys at medium- and high-dosed groups disappeared quickly. The effect of low-dosed group was equal to that of positive control group, acyclovir(ACV); the results of the titer tests showed that the virus decreased rapidly by using REE, especially in the medium- and high-dosed groups which were much more marked than the low-dosed group; Q test of the data showed that total mean value had statistical significance (F=49.1459, P<0.01); moreover there were statistical significance between therapy groups (ACV, DH1, DH2, DH3) and non-therapy groups (VC) (P<0.01 ) and between DH2, DH3 and DH1 (P<0.01); no statistical significance were found between DH1, DH2 or DH3 and ACV (P>0.05). Results show that as to the effect of decreasing the average of the total titer, rhubarb is as effective as ACV; furthermore, the medium- and high-dosed groups are superior to the low-dosed group.</p><p><b>CONCLUSIONS</b>REE has significant anti-viral effect on HSV in vivo; there will be a wide application foreground of it in clinical usage.</p>