[Objective] To investigate the mechanism of compound traditional Chinese medicine （TCM） on Newcastle disease virus （NDV） and to provide a scientific basis for the reasonable usage of antiviral drugs in clinic. [Me...[Objective] To investigate the mechanism of compound traditional Chinese medicine （TCM） on Newcastle disease virus （NDV） and to provide a scientific basis for the reasonable usage of antiviral drugs in clinic. [Method] The compound TCM was composed of Hedyotis diffusa, Lonicera japonica Thunb, Radix astragali and Glycyrrhiza uralensis. Different dilutions of fluid extract were prepared. Its antiviral effects on NDV were observed through three inoculation ways, first, inoculation with the medicine and NDV mixture which had been incubated at 37 ℃; second, incubating chicken embryo fibroblasts （CEF） with the medicine followed by inoculation with NDV; third, inoculation with N DV followed by incubating CEF with the medicine. The A,= was determined by M]-r [ 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） ~ method. Therapeutic indexes were used to evaluate the antiviral effects. [ Result] The minimum effective concentration of the compound TCM which acted through the three ways was 1.0 × 2^-10 1.0 × 2^-8 and 1.0 × 2^-7 g/ml, respectively. The antiviral effects of the compound TCM were the best through inoculation with the incubated medicine and NDV mixture, followed by the second method and the third method. [ Conclusion] The compound TCM can not only kill NDV directly in vitro but also inhibit viral propagation.展开更多
Objective: To explore the antitumor effects of hemaagglutinin-neuraminase gene (HN gene) from Newcastle disease virus. Methods: Plasmid vaccine of pIRHN was constructed and transfected into HeLa cells. The expression...Objective: To explore the antitumor effects of hemaagglutinin-neuraminase gene (HN gene) from Newcastle disease virus. Methods: Plasmid vaccine of pIRHN was constructed and transfected into HeLa cells. The expression of HN was analyzed by Western blot analysis, and the mode of cell death was detected by fluorescence microscope, gel electrophoresis and TUNEL assay and the expression of p53 and bcl-2 was also analyzed in transfected Hela cells. The effect of pIRHN on sialic acid contents in the Hela cell was examined. Results: pIRHN nucleic acid vaccines could be expressed in eukaryotic cell. pIRHN could induce apoptosis after HeLa cells were transfected. The effect of antitumor responses of pIRHN was correlated with the contents of sialic acid in tumor cells, and there was no prominent evidence for the relatedness of the antitumor effect with the expression of p53 and bcl-2. Conclusion: pIRHN may become a new antitumor biological agent.展开更多
Background and Aims:Hepatocellular carcinoma(HCC)is listed as one of the most common causes of cancer-related death.Oncolytic therapy has become a promising treatment because of novel immunotherapies and gene editing ...Background and Aims:Hepatocellular carcinoma(HCC)is listed as one of the most common causes of cancer-related death.Oncolytic therapy has become a promising treatment because of novel immunotherapies and gene editing technology,but biosafety concerns remain the biggest limitation for clinical application.We studied the the antitumor activity and biosafety of the wild-type Newcastle disease virus HK84 strain(NDV/HK84)and 10 other NDV strains.Meth-ods:Cell proliferation and apoptosis were determined by cell counting Kit-8 and fluorescein isothiocyanate Annexin V apoptosis assays.Colony formation,wound healing,and a xenograft mouse model were used to evaluate in vivo and in vitro oncolytic effectiveness.The safety of NDV/HK84 was tested in nude mice by an in vivo luciferase imaging system.The replication kinetics of NDV/HK84 in normal tis-sues and tumors were evaluated by infectious-dose assays in eggs.RNA sequencing analysis was performed to explore NDV/HK84 activity and was validated by quantitative real-time PCR.Results:The cell counting Kit-8 assays of vi-ability found that the oncolytic activity of the NDV strains differed with the multiplicity of infection(MOI).At an MOI of 20,the oncolytic activity of all NDV strains except the DK/JX/21358/08 strain was>80%.The oncolytic activities of the NDV/HK84 and DK/JX/8224/04 strains were>80%at both MOI=20 and MOI=2.Only NDV/HK84 had>80%oncolytic activities at both MOI=20 and MOI=2.We chose NDV/HK84 as the candidate virus to test the oncolytic effect of NDV in HCC in the in vitro and in vivo experiments.NDV/HK84 killed human SK-HEP-1 HCC cells without affecting healthy cells.Conclusions:Intratumor infection with NDV/HK84 strains compared with vehicle controls or positive controls indicated that NDV/HK84 strain specifically inhib-ited HCC without affecting healthy mice.High-throughput RNA sequencing showed that the oncolytic activity of NDV/HK84 was dependent on the activation of type I interferon signaling.展开更多
文摘[Objective] To investigate the mechanism of compound traditional Chinese medicine （TCM） on Newcastle disease virus （NDV） and to provide a scientific basis for the reasonable usage of antiviral drugs in clinic. [Method] The compound TCM was composed of Hedyotis diffusa, Lonicera japonica Thunb, Radix astragali and Glycyrrhiza uralensis. Different dilutions of fluid extract were prepared. Its antiviral effects on NDV were observed through three inoculation ways, first, inoculation with the medicine and NDV mixture which had been incubated at 37 ℃; second, incubating chicken embryo fibroblasts （CEF） with the medicine followed by inoculation with NDV; third, inoculation with N DV followed by incubating CEF with the medicine. The A,= was determined by M]-r [ 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） ~ method. Therapeutic indexes were used to evaluate the antiviral effects. [ Result] The minimum effective concentration of the compound TCM which acted through the three ways was 1.0 × 2^-10 1.0 × 2^-8 and 1.0 × 2^-7 g/ml, respectively. The antiviral effects of the compound TCM were the best through inoculation with the incubated medicine and NDV mixture, followed by the second method and the third method. [ Conclusion] The compound TCM can not only kill NDV directly in vitro but also inhibit viral propagation.
基金This work was supported by the National 973 Basic Research Development Program of China (No. G199011902).
文摘Objective: To explore the antitumor effects of hemaagglutinin-neuraminase gene (HN gene) from Newcastle disease virus. Methods: Plasmid vaccine of pIRHN was constructed and transfected into HeLa cells. The expression of HN was analyzed by Western blot analysis, and the mode of cell death was detected by fluorescence microscope, gel electrophoresis and TUNEL assay and the expression of p53 and bcl-2 was also analyzed in transfected Hela cells. The effect of pIRHN on sialic acid contents in the Hela cell was examined. Results: pIRHN nucleic acid vaccines could be expressed in eukaryotic cell. pIRHN could induce apoptosis after HeLa cells were transfected. The effect of antitumor responses of pIRHN was correlated with the contents of sialic acid in tumor cells, and there was no prominent evidence for the relatedness of the antitumor effect with the expression of p53 and bcl-2. Conclusion: pIRHN may become a new antitumor biological agent.
基金supported by research grants from the Guangdong Science and Technology Innovation Strategy Special Found(2019B121205009)the Guangdong Science and Technology Special Found(190830095586328 and 200109155890863)and the Li Ka Shing Foundation.
文摘Background and Aims:Hepatocellular carcinoma(HCC)is listed as one of the most common causes of cancer-related death.Oncolytic therapy has become a promising treatment because of novel immunotherapies and gene editing technology,but biosafety concerns remain the biggest limitation for clinical application.We studied the the antitumor activity and biosafety of the wild-type Newcastle disease virus HK84 strain(NDV/HK84)and 10 other NDV strains.Meth-ods:Cell proliferation and apoptosis were determined by cell counting Kit-8 and fluorescein isothiocyanate Annexin V apoptosis assays.Colony formation,wound healing,and a xenograft mouse model were used to evaluate in vivo and in vitro oncolytic effectiveness.The safety of NDV/HK84 was tested in nude mice by an in vivo luciferase imaging system.The replication kinetics of NDV/HK84 in normal tis-sues and tumors were evaluated by infectious-dose assays in eggs.RNA sequencing analysis was performed to explore NDV/HK84 activity and was validated by quantitative real-time PCR.Results:The cell counting Kit-8 assays of vi-ability found that the oncolytic activity of the NDV strains differed with the multiplicity of infection(MOI).At an MOI of 20,the oncolytic activity of all NDV strains except the DK/JX/21358/08 strain was>80%.The oncolytic activities of the NDV/HK84 and DK/JX/8224/04 strains were>80%at both MOI=20 and MOI=2.Only NDV/HK84 had>80%oncolytic activities at both MOI=20 and MOI=2.We chose NDV/HK84 as the candidate virus to test the oncolytic effect of NDV in HCC in the in vitro and in vivo experiments.NDV/HK84 killed human SK-HEP-1 HCC cells without affecting healthy cells.Conclusions:Intratumor infection with NDV/HK84 strains compared with vehicle controls or positive controls indicated that NDV/HK84 strain specifically inhib-ited HCC without affecting healthy mice.High-throughput RNA sequencing showed that the oncolytic activity of NDV/HK84 was dependent on the activation of type I interferon signaling.
