RESUMEN
The aim of the study was to successfully construct three plasmids, which include the GALV.fus gene plasmid regulated by the herpes simplex virus type 1 (HSV-1) late expression gene-UL38 promoter and induced by HSV-1 (HSV-UL38P-GALV.fus), the cytomegalovirus promoter without tumor specificity (CMVP) GALV.fus plasmid (HSV-CMVP-GALV.fus), and the control plasmid in which the GALV.fus gene fragment was replaced by the enhanced green fluorescent protein (EGFP) gene fragment (HSV-CMVP-EGFP). The three constructed plasmids were all packaged and named as Synco-2, Synco-1, and Baco-1. The plasmids were amplified in coliform bacterium and transfected into Vero cells using lipofectamine. These recombinant HSV-1 were amplified in Vero cells and purified by conventional methods of cesium chloride, TCID50 method is used to measure virus titers. The total RNA was then extracted from the HepG2 cells transfected by Synco-1 and Synco-2, and the expression of GALV.fus mRNA was detected by RT-PCR. The three recombinant HSV-1 vectors were propagated in Vero cells and purified by cesium chloride density gradient centrifugation, titrated by TCID50 method, and packaged. The titers of Baco-1, Synco-1, and Synco-2 were 3 × 10(10), 1 × 10(11), and 4 × 10(10) pfu/ml. The GALV.fus gene was identified in the infected HepG2 cells by RT-PCR method.
Asunto(s)
ADN Recombinante/genética , Ingeniería Genética/métodos , Herpesvirus Humano 1/genética , Virus de la Leucemia del Gibón/genética , Técnicas de Amplificación de Ácido Nucleico , Proteína FUS de Unión a ARN/genética , Ensamble de Virus , Animales , Chlorocebus aethiops , Terapia Genética , Células Hep G2 , Herpesvirus Humano 1/fisiología , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia , Plásmidos/genética , ARN/genética , ARN/aislamiento & purificación , Transfección , Células VeroRESUMEN
Objective To investigate the antitumor effect of special promoter-controlled Gibbon ape leukemia virus membrane fusion glycoprotein (GALV.fus) mediated by type Ⅰ herpes simplex virus (HSV-Ⅰ) on lung adenocarcinoma. Methods Recombinant HSV-Ⅰ plasmids encoding GALV.fus was introduced into green monkey kidney cells(Vero)by liposome to amplify the virus, and then the virus was transfected into lung adenocarcinoma (A549), human fetal fibroblasts (HFL-Ⅰ GNHu 5) and human lung adenocarcinoma xenografts which were established in nude mice subcutaneously to observe antitumor and cytotoxic effect in vitro and in vivo; Recombined cytomegalovirus (CMV) containing GALV.fus or enhanced green fluorescence protein were served as control. Results Recombinant HSV-Ⅰ virus were packed successfully. Heterotransplantative tumourigenicity of the tumour was 100% in nude mice after A549 cells were inoculated. Recombinant HSV-Ⅰvirus exerted obvious antitumor effects in vitro, with relative survival rate of 23%, while for CMV virus containing GALV. fus, the rate was 20%, and for CMV virus encoding EGFP, the rate was 68%. Recombinant HSV-Ⅰvirus also showed striking antitumor effect on the implanted tumor. Conclusion GALV.fus has powerful effect against lung cancer in vitro and in vivo and maybe a promising candidate for gene therapy.
RESUMEN
Image analysis plays an important role in morphological study of tissues and cells and their quantitative analysis. It also contributes to clinical pathological diagnosis. With the rapid development of computer technology great progress has been made in the image analysis system, its measurement, rapidity, accuracy and the extent of automation have been greatly enhanced. In the meantime advances in medicine give impetus to its improvement. In this paper, the process of development, the basic structure and mechanism of image analysis system are discussed and the application of image analysis system to the study of biomedicinal is presented.
Asunto(s)
Procesamiento de Imagen Asistido por Computador , Métodos , MedicinaRESUMEN
Objective To explore the digital morphological and chromatic characteristics of erythrocytes infected with Plasmodium vivax. Methods The images of both normal and P. vivax infected erythrocytes were segmented and measured for morphologic parameters including area, length, breadth, perimeter, roundness, aspect ratio, equivalent circle diameter, as well as chromatic parameters including saturation and color (red, green, blue). A statistic analysis was performed for these parameters. Results Both morphological and chromatic parameters showed high significant differences between the normal and the infected erythrocytes, and high significant differences between the normal and the erythrocytes infected with different stages of P. vivax. Conclusion The differences mentioned above could be used as the basis for automatic identification of P.vivax in thin peripheral blood smears.