ABSTRACT
This study performed phylogenic analysis on a dengue strain isolated from an outbreak of dengue fever in 2009 at Yiwu City of Zhejiang Province,China,and further to analyze the immunogenicity of E protein of this viral isolate.Firstly,the viral genome was amplified by RT-PCR and phylogenetic trees were constructed by MEGA 4 based on both nucleotide and amino acid sequences of E and NS1 proteins.The phylogenetic analysis showed that the similarity of Yiwu strain with the Guangzhou GZ1D3 strain and the India GWL-25 strain was over 99％.Secondly,the expression plasmid of E protein was constructed and transfected into 293T cells.The secreted E protein were then purified by sucrose density gradient centrifugation and used to inoculate BALB/c mice.The humoral immunity was evaluated by ELISA and neutralizing antibody analysis.Resuits showed that the E protein of Yiwu strain could induce dengue specific IgG antibodies and neutralizing antibodies.Therefore,the study found that the Yiwu strain was classified into the subtype Ⅲ of dengue virus type 3 (DENV-3),and the E protein of this strain had strong immunogenicity
ABSTRACT
AIM: To investigate the mechanism of berberine’s inhibiting growth and metastasis of tumor by observing the effects of berberine on proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were incubated with berberine. MTT assay and quantitative immunocytochemistry were used to detect cell proliferation and the expression of proliferation cell nuclear antigen (PCNA). Morphologic changes of apoptosis were observed by fluorescent staining, and Rhodamin123 was used to determinate mitochondrial membrane potential under the laser confocal microscopy. RESULTS: HUVEC proliferation was inhibited by co-incubating with berberine (20 mg/L) for 24 h and berberine (10 mg/L, 20 mg/L) for 48 h (P