RESUMO
AIM: To evaluate the efficiency of 3 short hairpin RNA (shRNA) interfering with the herpes simplex virus type 1 (HSV-1) gene coding glycoprotein D (gD) for inhibiting the gD expression and virus replication in vitro. METHODS: Vero cells were selected for an in vitro model of infection. Three shRNA sequences (shRNAgD1, -gD2, and -gD3) targeting specifically the gD gene of HSV-1 were selected for evaluating the antiviral effects. The antiviral effects of shRNA in the cells infected with HSV-1 were evaluated by cytopathic effect (CPE) observations and plaque assays. The transcription level of viral RNA and the gD expression were studied by RT-PCR, Western blotting, and flow cytometry. RESULTS: With the 3 shRNA at a final concentration of 120 nmol/L, a significant inhibition of CPE in the HSV-1-infected cells was observed. The ED50 of shRNA-gD1, gD2, and gD3 were 48.74+/-2.57, 57.13+/-3.24, and 114.64+/-5.12 nmol/L, respectively. The gD gene decreased significantly after viral infection in the Vero cells pretreated with shRNA compared to the virus group. The expressions of the gD protein, determined by Western blotting and flow cytometry, were also drastically decreased in shRNA-transfected cells. CONCLUSION: Exogenous shRNA molecules can suppress the HSV-1 gD expression. They are inhibitors of HSV replication during infection in Vero cells.
Assuntos
Regulação Viral da Expressão Gênica/fisiologia , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/fisiologia , RNA Viral/fisiologia , Animais , Chlorocebus aethiops , Efeito Citopatogênico Viral/efeitos dos fármacos , Regulação Viral da Expressão Gênica/genética , Sequências Repetidas Invertidas/genética , RNA Viral/biossíntese , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção , Células Vero , Proteínas do Envelope Viral/biossíntese , Proteínas do Envelope Viral/genética , Replicação Viral/efeitos dos fármacosRESUMO
OBJECTIVE: To understand the status of coxsackievirus B (CVB) infection in senior subjects and explore the association of the infection with cardiovascular diseases. METHOD: By means of ELISA and RT-PCR, CVB IgG antibody and nucleic acid were detected in 172 patients with cardiopathy and 58 control subjects. RESULTS: The positivity rate of CVB IgG antibody was 43.61% (75/172) in the patient group and 15.52% (9/58) in the control group, showing obvious difference between the two groups. CVB RNA tests revealed a CVB RNA-positivity rate of 20.93% (36/172) in the former group and 3.45% (2/58) in the latter, with also a significant difference (P<0.01). CONCLUSION: CVB infection is prevalent in aged patients with heart disease, which should be given due attention to.
