Canine parvovirus(CPV)is a high morbidity and lethality virus which causes severe enteric disease in dogs.In an attempt to gain canine-origin neutralizing antibodies against CPV,single chain variable fragment(scFv)bac...Canine parvovirus(CPV)is a high morbidity and lethality virus which causes severe enteric disease in dogs.In an attempt to gain canine-origin neutralizing antibodies against CPV,single chain variable fragment(scFv)bacteria display libraries against the protective antigen VP2 were constructed and screened.VP2 specific scFvs were selected following three rounds of screening procedures.Selected scFvs were characterized by FCM and ELISA.Seven scFvs showed high affinity and specific binding to CPV.Moreover,the neutralizing activity of the antibody was preliminarily identified by CPV(100 TCID_(50))in vitro and scFv-2 showed the neutralizing ability with a titer of 32768.This study generated the first canine-origin neutralizing scFv against CPV.The neutralizing scFv would be constructed into full-length antibody in the future.This study laid the foundation for the generation of an effective therapeutic reagent with long half-life and no immunological rejection for the prevention and treatment of CPV infection.展开更多
Goose parvovirus(GPV)can cause a highly contagious and fatal gosling plague(GP)disease in goslings and muscoy ducklings.Here,three goose-origin neutralizing single chain variable fragment(scFv)antibodies against GPV S...Goose parvovirus(GPV)can cause a highly contagious and fatal gosling plague(GP)disease in goslings and muscoy ducklings.Here,three goose-origin neutralizing single chain variable fragment(scFv)antibodies against GPV SYG-61 were isolated.The genes of scFv antibodies were derived from goslings immunized with GPV SYG-61,and scFvs were subcloned into a pBSD vector for the construction of pBSD-scFv libraries.The pBSD-scFv libraries were screened following three rounds using VP2(protective antigen of GPV)as the bait by flow cytometry(FCM).After screening,the 15 clones with high mean fluorescence intensity(MFI)were isolated and sequenced.These 15 scFvs were expressed by pET-28a(+)in E.coli.The specificity and affinity of the 15 purified scFvs were successfully confirmed by ELISA.In the preliminary neutralization experiment on primary goose embryo fibroblast(GEF)in vitro,three of the 15 purified scFvs(named scFv-10,scFv-11 and scFv-50)showed significant neutralizing capacities.The study generated the first goose-origin neutralizing scFv against GPV and laid the foundation for the appearance of full-length goose-origin neutralizing monoclonal antibody against GPV.展开更多
Recombinant Newcastle disease virus(rNDV)has shown an anti-cancer effect in preclinical studies,but has never been tested for lung cancer models.This study explored the anti-cancer activity of genetically modified NDV...Recombinant Newcastle disease virus(rNDV)has shown an anti-cancer effect in preclinical studies,but has never been tested for lung cancer models.This study explored the anti-cancer activity of genetically modified NDV expressing IL-2(rNDV–IL-2)in lung cancer models.This study used Lewis lung carcinoma cell line(LLC)to create tumor models in C57 female mice,the tumor-bearing mice were treated with rNDV-IL-2,rNDV and phosphate-buffered saline(PBS),respectively.In vitro results revealed that rNDV effectively infected malignant cells and expressed IL-2,in vivo results revealed that rNDV expressing IL-2 was highly efficient in inhibiting lung cancer tumors,with an average tumor size of 291.255 mm^3 in rNDV-IL-2 group compared to 763.068 mm^3 in rNDV group and 1101.68 mm^3 in PBS group.For the survival studies,treatment with rNDV-IL-2 enhanced the survival rates of tumor-bearing mice by 36%compared to those of rNDV treated mice and by 80%compared to those of vehicle-treated mice(survival rate:12 out of 15 for rNDV-IL-2 group;seven out of 15 for rNDV group and zero out of 15 for vehicle group).These results demonstrated that rNDV-expressed IL-2 enhanced the intrinsic anti-tumor ability of Newcastle disease virus in lung cancer models by further restrain of lung tumor growth and improvement of the survival rates of the tumor-bearing mice.The genetically modified rNDV-IL-2 was a good candidate for lung cancer therapy.展开更多
绿色环境效率反映经济发展与资源环境之间的平衡关系,合理评价资源型城市的环境效率对于促进经济高质量发展具有重要的现实意义。首先从异质性环境污染排放角度出发,选取了四种环境压力指标,采用非径向的Slacks Based Model(SBM)模型和S...绿色环境效率反映经济发展与资源环境之间的平衡关系,合理评价资源型城市的环境效率对于促进经济高质量发展具有重要的现实意义。首先从异质性环境污染排放角度出发,选取了四种环境压力指标,采用非径向的Slacks Based Model(SBM)模型和Sequential Malmquist指数模型,对山西省2003—2016年11个地级市的环境效率和环境生产率的区域差异及其动态演进进行了深入分析,然后采用固定效应模型、差分广义矩估计模型和面板分位数回归模型分析了环境规制对山西环境生产率的影响。研究结果表明:(1)在维持现有技术水平不变的前提下,山西省整体的环境效率仍有25.31%的改进潜力;中部地区的环境效率值高于北部与南部地区,但是三大区域的环境效率值都呈下降趋势。(2)山西省环境生产率平均每年提高14.63%,其中技术效率对环境生产率的增长表现为负效应,而技术进步是环境生产率提高的主要来源。(3)环境规制与环境生产率之间并不存在正向或者负向的线性关系,而是存在显著的“U”型关系;不同分位点上环境规制对环境生产率变动的影响表现出明显的异质性。展开更多
基金Supported by the National Key R&D Program of China(2017YFD0501102,2016YFD0501003)Science and Technology Department of Heilongjiang Province(GX18B018)Education Department of Heilongjiang Province(TSTAU-R2018017)。
文摘Canine parvovirus(CPV)is a high morbidity and lethality virus which causes severe enteric disease in dogs.In an attempt to gain canine-origin neutralizing antibodies against CPV,single chain variable fragment(scFv)bacteria display libraries against the protective antigen VP2 were constructed and screened.VP2 specific scFvs were selected following three rounds of screening procedures.Selected scFvs were characterized by FCM and ELISA.Seven scFvs showed high affinity and specific binding to CPV.Moreover,the neutralizing activity of the antibody was preliminarily identified by CPV(100 TCID_(50))in vitro and scFv-2 showed the neutralizing ability with a titer of 32768.This study generated the first canine-origin neutralizing scFv against CPV.The neutralizing scFv would be constructed into full-length antibody in the future.This study laid the foundation for the generation of an effective therapeutic reagent with long half-life and no immunological rejection for the prevention and treatment of CPV infection.
基金Supported by the National Key R&D Program of China(2017YFD0501102,20I7YFD050I103-03 and 2017YFD0501004)Science and Technology Department of Heilongjiang Province(GX18B018)Education Department ofHeilongjiang Province(TSTAU-R2018017)。
文摘Goose parvovirus(GPV)can cause a highly contagious and fatal gosling plague(GP)disease in goslings and muscoy ducklings.Here,three goose-origin neutralizing single chain variable fragment(scFv)antibodies against GPV SYG-61 were isolated.The genes of scFv antibodies were derived from goslings immunized with GPV SYG-61,and scFvs were subcloned into a pBSD vector for the construction of pBSD-scFv libraries.The pBSD-scFv libraries were screened following three rounds using VP2(protective antigen of GPV)as the bait by flow cytometry(FCM).After screening,the 15 clones with high mean fluorescence intensity(MFI)were isolated and sequenced.These 15 scFvs were expressed by pET-28a(+)in E.coli.The specificity and affinity of the 15 purified scFvs were successfully confirmed by ELISA.In the preliminary neutralization experiment on primary goose embryo fibroblast(GEF)in vitro,three of the 15 purified scFvs(named scFv-10,scFv-11 and scFv-50)showed significant neutralizing capacities.The study generated the first goose-origin neutralizing scFv against GPV and laid the foundation for the appearance of full-length goose-origin neutralizing monoclonal antibody against GPV.
基金Supported by the National Key R&D Program of China(2017YFD0501102)the National Natural Science Foundation of China(31200121)。
文摘Recombinant Newcastle disease virus(rNDV)has shown an anti-cancer effect in preclinical studies,but has never been tested for lung cancer models.This study explored the anti-cancer activity of genetically modified NDV expressing IL-2(rNDV–IL-2)in lung cancer models.This study used Lewis lung carcinoma cell line(LLC)to create tumor models in C57 female mice,the tumor-bearing mice were treated with rNDV-IL-2,rNDV and phosphate-buffered saline(PBS),respectively.In vitro results revealed that rNDV effectively infected malignant cells and expressed IL-2,in vivo results revealed that rNDV expressing IL-2 was highly efficient in inhibiting lung cancer tumors,with an average tumor size of 291.255 mm^3 in rNDV-IL-2 group compared to 763.068 mm^3 in rNDV group and 1101.68 mm^3 in PBS group.For the survival studies,treatment with rNDV-IL-2 enhanced the survival rates of tumor-bearing mice by 36%compared to those of rNDV treated mice and by 80%compared to those of vehicle-treated mice(survival rate:12 out of 15 for rNDV-IL-2 group;seven out of 15 for rNDV group and zero out of 15 for vehicle group).These results demonstrated that rNDV-expressed IL-2 enhanced the intrinsic anti-tumor ability of Newcastle disease virus in lung cancer models by further restrain of lung tumor growth and improvement of the survival rates of the tumor-bearing mice.The genetically modified rNDV-IL-2 was a good candidate for lung cancer therapy.
文摘绿色环境效率反映经济发展与资源环境之间的平衡关系,合理评价资源型城市的环境效率对于促进经济高质量发展具有重要的现实意义。首先从异质性环境污染排放角度出发,选取了四种环境压力指标,采用非径向的Slacks Based Model(SBM)模型和Sequential Malmquist指数模型,对山西省2003—2016年11个地级市的环境效率和环境生产率的区域差异及其动态演进进行了深入分析,然后采用固定效应模型、差分广义矩估计模型和面板分位数回归模型分析了环境规制对山西环境生产率的影响。研究结果表明:(1)在维持现有技术水平不变的前提下,山西省整体的环境效率仍有25.31%的改进潜力;中部地区的环境效率值高于北部与南部地区,但是三大区域的环境效率值都呈下降趋势。(2)山西省环境生产率平均每年提高14.63%,其中技术效率对环境生产率的增长表现为负效应,而技术进步是环境生产率提高的主要来源。(3)环境规制与环境生产率之间并不存在正向或者负向的线性关系,而是存在显著的“U”型关系;不同分位点上环境规制对环境生产率变动的影响表现出明显的异质性。
