The present study was designed to evaluate the protective effects of Reduning injection against Enterovirus 71(EV71) in Vero cells and in mice. The Vero cells were infected with 100 and 50 TCID50(50% tissue culture in...The present study was designed to evaluate the protective effects of Reduning injection against Enterovirus 71(EV71) in Vero cells and in mice. The Vero cells were infected with 100 and 50 TCID50(50% tissue culture infective dose) of EV71, respectively. The inhibition of Reduning injection on cytopathic effect(CPE) was detected. Meanwhile, a mouse model produced by intraperitoneal EV71-infection(106 TCID50), was used to investigate the protective effects of Reduning injection. The total survival rate, living time, daily survival rate, weight ratio, and score for symptoms were examined. The viral loads in Vero cells and muscle tissues were detected using real-time PCR. Finally, the content of cytokines was analyzed by ELISA. In the Vero cells, 2.5 mg crude drug·m L-1 of Reduning injection inhibited CPE induced by EV71 infection. In the mice, 1.3 g crude drug·kg-1 of Reduning injection rescued death triggered by infection, in comparison with model group. Moreover, the survival rate, weight ratio, and clinical scores were also improved. The viral RNA copies in the Vero cells and the mice muscle tissues were reduced. Besides, the steep EV71-induced accumulations of TNF-α and MCP-1 were decreased by Reduning injection. In conclusion, Reduning injection showed promising protective effects against EV71 in Vero cells and in mice.展开更多
RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequenc...RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequence. dsRNAmediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression.Synthetic 21-23 nucleotide (nt) small interfering RNA (siRNA) with 2 nt 3' overhangs was recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we studied the effects of synthetic siRNA duplexes targeted to SARS coronavirus structural proteins E, M, and N in a cell culture system. Among total 26 siRNA duplexes, we obtained 3 siRNA duplexes which could sequence-specifically reduce target genes expression over 80％ at the concentration of 60 nM in Vero E6 cells. The downregulation effect was in correlation with the concentrations of the siRNA duplexes in a range of 0～60 nM. Our results also showed that many inactive siRNA duplexes may be brought to life simply by unpairing the 5' end of the antisense strands. Results suggest that siRNA is capable of inhibiting SARS coronavirus genes expression and thus may be a new therapeutic strategy for treatment of SARS.展开更多
OBJECTIVE: To evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application...OBJECTIVE: To evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases. METHODS: Vero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells. RESULTS: Five pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect. CONCLUSION: siRNA may be effective in inhibiting SARS-associated coronavirus replication.展开更多
In this study,we have investigated the antiviral activity of GuiQi polysaccharides (GQP) upon enterovirus 71 (EV71) in vitro.An assay using methyl thiazolyl tetrazolium (MTT),and analyses of cytopathic effects (CPE)we...In this study,we have investigated the antiviral activity of GuiQi polysaccharides (GQP) upon enterovirus 71 (EV71) in vitro.An assay using methyl thiazolyl tetrazolium (MTT),and analyses of cytopathic effects (CPE)were used to examine the antiviral activity of GQP upon Vero cells infected with EV71.The results revealed that GQP at concentrations below 31.2μg/mL exhibited significant antiviral effects upon EV71 when applied under three different experimental protocols.GQP was most strongly active in preventing the adsorption of EV71 to target cells and in this respect it was significantly more effective than ribavirin.In addition,it was clear that GQP could inhibit viral replication when added to cells 2 h after infection,but if added at the point of infection its effect was weak.GQP is considered to be less toxic than ribavirin,and may warrant further evaluation as a possible agent in the treatment of hand,foot and mouth disease (HFMD).展开更多
Due to the insufficient supply of embryonated chicken eggs,the preparation of large quantities of inactivated influenza vaccines will require an alternative virus culture system after the emergence or reemergence of a...Due to the insufficient supply of embryonated chicken eggs,the preparation of large quantities of inactivated influenza vaccines will require an alternative virus culture system after the emergence or reemergence of a pandemic influenza virus.The Vero cell is one of the ideal options since it was used for producing many kinds of human vaccines.However,most of the influenza viruses can not grow well in Vero cells.To develop a new influenza vaccine with Vero cells as a substrate,the virus needs to adapt to this cell substrate to maintain high growth characteristics.By serial passages in Vero cells,the B/Yunnan/2/2005va(B)strain was successfully adapted to Vero cells,with the hemagglutination titer(HAT)of the virus reaching 1:512.The high growth characteristic of this strain is stable up to 21 passages.The strain was identified by hemagglutination inhibition (HAI)test and sequencing respectively;the HA1 gene sequence of the virus was cloned and analyzed.The screening and establishment of high growth B virus provides an important tool for influenza vaccine production in Vero cells.展开更多
The purpose of this study was to investigate the effect of curcumin-treated Herpes simplex virus-1 (HSV-1) and Herpes simplex virus-2 (HSV-2) virions in cultured Vero cells. Previous studies have indicated that curcum...The purpose of this study was to investigate the effect of curcumin-treated Herpes simplex virus-1 (HSV-1) and Herpes simplex virus-2 (HSV-2) virions in cultured Vero cells. Previous studies have indicated that curcumin, a polyphenol extracted from the plant Curcuma longa, has demonstrated antiviral properties against a variety of viruses. After establishing the maximum non-cytotoxic concentrations of curcumin on Vero cells, HSV-1 and HSV-2 virions were treated with varying concentrations of curcumin. The effect on infectivity was determined by antiviral assays, using WST-1, plaque assays, adsorption and penetration assays. Treating HSV-1 and HSV-2 viruses with curcumin, at a concentration of 30 μM, reduces the production of infectious HSV-1 and HSV-2 virions in cultured Vero cells by interfering with the adsorption process. These results support the potential of curcumin to be used as a therapeutic agent to reduce the transmission of HSV-1 and HSV-2.展开更多
文摘目的 评价不同细胞基质制备的肠道病毒71型（EV71）灭活疫苗上市后常规免疫安全性。 方法 EV71灭活疫苗疑似预防接种异常反应（AEFI）数据来源于中国免疫规划信息管理系统，为2017年接种EV71 灭活疫苗后江苏省报告的监测数据；受种儿童人口学信息和接种剂次数来源于江苏省预防接种信息管理系统。 结果 2017年江苏省共接种EV71灭活疫苗316 889剂，报告AEFI 208例，报告发生率为65.64/10万剂（95% CI ：56.72/10万剂～74.56/10万剂），其中一般反应55.54/10万剂（95% CI ：47.34/10万剂～63.74/10万剂），异常反应 7.89/10万剂（95% CI ：4.80/10万剂～10.98/10万剂）；不同细胞基质的疫苗AEFI发生率（χ 2 =35.61， P ＜0.01）和一般反应发生率（χ 2 =34.09， P ＜0.01）差异有统计学意义，异常反应发生率差异无统计学意义（χ 2 =1.88， P =0.17），均未见严重异常反应，208例AEFI均未住院且均痊愈。EV71灭活疫苗（Vero细胞）首剂接种一般反应发生率高于第二剂（χ 2 =6.94， P ＜0.01），不同季度接种EV71灭活疫苗（人二倍体细胞）一般反应发生率差异有统计学意义（χ 2 =18.86， P ＜0.01）。 结论 不同细胞基质的EV71灭活疫苗在常规免疫接种中AEFI发生率均不高，有良好的安全性；一般反应以发热为主，异常反应以过敏性-荨麻疹、过敏性皮疹和过敏性斑丘疹等过敏性反应为主。
基金supported by National Major New Drugs Innovation and Development Project from Ministry of Science and Technology of China(2013ZX09402203)
文摘The present study was designed to evaluate the protective effects of Reduning injection against Enterovirus 71(EV71) in Vero cells and in mice. The Vero cells were infected with 100 and 50 TCID50(50% tissue culture infective dose) of EV71, respectively. The inhibition of Reduning injection on cytopathic effect(CPE) was detected. Meanwhile, a mouse model produced by intraperitoneal EV71-infection(106 TCID50), was used to investigate the protective effects of Reduning injection. The total survival rate, living time, daily survival rate, weight ratio, and score for symptoms were examined. The viral loads in Vero cells and muscle tissues were detected using real-time PCR. Finally, the content of cytokines was analyzed by ELISA. In the Vero cells, 2.5 mg crude drug·m L-1 of Reduning injection inhibited CPE induced by EV71 infection. In the mice, 1.3 g crude drug·kg-1 of Reduning injection rescued death triggered by infection, in comparison with model group. Moreover, the survival rate, weight ratio, and clinical scores were also improved. The viral RNA copies in the Vero cells and the mice muscle tissues were reduced. Besides, the steep EV71-induced accumulations of TNF-α and MCP-1 were decreased by Reduning injection. In conclusion, Reduning injection showed promising protective effects against EV71 in Vero cells and in mice.
基金supported by the Grant No.2003AA208215 from the National High Technology Programs of Chinathe Grant No.30270311 from the National Natural Science Foundation of China.
文摘RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequence. dsRNAmediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression.Synthetic 21-23 nucleotide (nt) small interfering RNA (siRNA) with 2 nt 3' overhangs was recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we studied the effects of synthetic siRNA duplexes targeted to SARS coronavirus structural proteins E, M, and N in a cell culture system. Among total 26 siRNA duplexes, we obtained 3 siRNA duplexes which could sequence-specifically reduce target genes expression over 80％ at the concentration of 60 nM in Vero E6 cells. The downregulation effect was in correlation with the concentrations of the siRNA duplexes in a range of 0～60 nM. Our results also showed that many inactive siRNA duplexes may be brought to life simply by unpairing the 5' end of the antisense strands. Results suggest that siRNA is capable of inhibiting SARS coronavirus genes expression and thus may be a new therapeutic strategy for treatment of SARS.
文摘OBJECTIVE: To evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases. METHODS: Vero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells. RESULTS: Five pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect. CONCLUSION: siRNA may be effective in inhibiting SARS-associated coronavirus replication.
基金supported by research grants from The National Natural Science Foundation of China(NO81260070)The Project of Science and Technology of Lanzhou(NO 2011-1-71)The Doctor Project of Lanzhou University of Technology(NO 0908ZXC127)
文摘In this study,we have investigated the antiviral activity of GuiQi polysaccharides (GQP) upon enterovirus 71 (EV71) in vitro.An assay using methyl thiazolyl tetrazolium (MTT),and analyses of cytopathic effects (CPE)were used to examine the antiviral activity of GQP upon Vero cells infected with EV71.The results revealed that GQP at concentrations below 31.2μg/mL exhibited significant antiviral effects upon EV71 when applied under three different experimental protocols.GQP was most strongly active in preventing the adsorption of EV71 to target cells and in this respect it was significantly more effective than ribavirin.In addition,it was clear that GQP could inhibit viral replication when added to cells 2 h after infection,but if added at the point of infection its effect was weak.GQP is considered to be less toxic than ribavirin,and may warrant further evaluation as a possible agent in the treatment of hand,foot and mouth disease (HFMD).
基金National"863 Project"(2006AA02Z409)National health project(200802023)Yunnan sciences and technology cooperation(2006XY29)
文摘Due to the insufficient supply of embryonated chicken eggs,the preparation of large quantities of inactivated influenza vaccines will require an alternative virus culture system after the emergence or reemergence of a pandemic influenza virus.The Vero cell is one of the ideal options since it was used for producing many kinds of human vaccines.However,most of the influenza viruses can not grow well in Vero cells.To develop a new influenza vaccine with Vero cells as a substrate,the virus needs to adapt to this cell substrate to maintain high growth characteristics.By serial passages in Vero cells,the B/Yunnan/2/2005va(B)strain was successfully adapted to Vero cells,with the hemagglutination titer(HAT)of the virus reaching 1:512.The high growth characteristic of this strain is stable up to 21 passages.The strain was identified by hemagglutination inhibition (HAI)test and sequencing respectively;the HA1 gene sequence of the virus was cloned and analyzed.The screening and establishment of high growth B virus provides an important tool for influenza vaccine production in Vero cells.
文摘The purpose of this study was to investigate the effect of curcumin-treated Herpes simplex virus-1 (HSV-1) and Herpes simplex virus-2 (HSV-2) virions in cultured Vero cells. Previous studies have indicated that curcumin, a polyphenol extracted from the plant Curcuma longa, has demonstrated antiviral properties against a variety of viruses. After establishing the maximum non-cytotoxic concentrations of curcumin on Vero cells, HSV-1 and HSV-2 virions were treated with varying concentrations of curcumin. The effect on infectivity was determined by antiviral assays, using WST-1, plaque assays, adsorption and penetration assays. Treating HSV-1 and HSV-2 viruses with curcumin, at a concentration of 30 μM, reduces the production of infectious HSV-1 and HSV-2 virions in cultured Vero cells by interfering with the adsorption process. These results support the potential of curcumin to be used as a therapeutic agent to reduce the transmission of HSV-1 and HSV-2.
