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Frontiers of Agricultural Science and Engineering 2015年第3期2卷 216-222页

作者： Yuzhou wang tongyan wang He YAN Fanli YANG Linghua GUO Qingyuan YANG Xule HU Feifei TAN Yan XIAO Xiangdong LI Kegong TIAN National Research Center for Veterinary Medicine Luoyang 471003China College of Animal Science and Veterinary Medicine Henan Agricultural UniversityZhengzhou 450002China

Pseudorabies(PR)is a devastating viral disease which leads to fatal encephalitis and respiratory disorders in pigs.Commercial gE-deleted live pseudorabies virus(PRV)vaccine has been widely used to control this disease in China.However,the new-emerging variants of PRV compromises the protection provided by current vaccines and lead to the outbreak of PR in vaccinated pig herds.Several killed and live vaccine candidates based on current PRV variants have been reported to be effective to control the disease.A subunit vaccine based on gB protein,one major PRV glycoprotein which elicits strong humoral and cellular immune responses,however,was never evaluated for protection against the current circulating PRV variants.In this study,full-length PRV gB protein was successfully expressed in baculovirus/insect cells in the soluble format and was tested on 3-week-old piglets as a subunit vaccine.Compared with unvaccinated pigs,the gB-vaccinated pigs developed specific antibody-mediated responses and were protected from the virulent PRV HN1201 challenge.All vaccinated pigs survived without showing any PRV-specific respiratory and neurological signs,but all unvaccinated pigs died within 7 days after HN1201 challenge.Hence,this novel gB-based vaccine could be applied as an effective subunit vaccine to control PRV variant in China.

关键词： pseudorabies virus glycoprotein B protein subunit vaccine

表达PEDV S蛋白重组PRV的构建与生物学特性评价

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中国兽医学报 2024年第4期44卷 657-663页

作者：侯晓璇穆永王同燕颜世君王孟月谭菲菲田克恭河南农业大学动物医学学院河南郑州450046 国家兽用药品工程技术研究中心河南洛阳471000 普莱柯生物工程股份有限公司河南洛阳471000

将猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)流行毒株的S胞外区基因克隆至含有gG基因上、下游同源臂以及抗性筛选基因的转移质粒pUC-ABK中,构建供体质粒pUC-ABK-S,获得线性同源重组片段AS-K-B。利用Red同源重组技术将线... 详细信息

将猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)流行毒株的S胞外区基因克隆至含有gG基因上、下游同源臂以及抗性筛选基因的转移质粒pUC-ABK中,构建供体质粒pUC-ABK-S,获得线性同源重组片段AS-K-B。利用Red同源重组技术将线性同源重组片段电转化至含有pPRVBac-GFPTK-gE-gI-11k-28k-的感受态中获得重组pPRVBac-S,最后拯救获得重组病毒rPRV-S。PCR及IFA鉴定结果显示,S胞外区基因成功转录且表达,生物学特性评价结果表明重组病毒能够稳定遗传,为研制猪流行性腹泻病毒活载体疫苗提供了思路。

关键词： PEDV S蛋白 Red同源重组重组PRV

RR1 and RR2 gene deletion affects the immunogenicity of a live attenuated pseudorabies virus vaccine candidate in natural pig host

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Frontiers of Agricultural Science and Engineering 2016年第1期3卷 81-86页

作者： Shijun YAN He YAN Chaolin ZHANG tongyan wang Qingyuan YANG Zhe SUN Yan XIAO Feifei TAN Xiangdong LI Kegong TIAN National Research Center for Veterinary Medicine College of Animal Science and Veterinary Medicine Henan Agricultural University

As virulence-determining genes, RR1 and RR2 encode the small subunit and large subunit of viral ribonucleotide reductase(RR) in pseudorabies virus which have been extensively studied in mice. However,their role in pigs has not been adequately investigated. In this study, we deleted RR1 and RR2 genes based on a TK/g E/g I triple gene-deleted pseudorabies virus and tested its efficacy in pigs as a vaccine candidate. The rescued virus showed similar growth properties and plaque size in vitro as its parent strain. In an animal study, the virus could elicit humoral immune responses shown by generation of g B-specific antibodies and virus neutralizing antibodies.However, vaccination could not provide protection against virulent pseudorabies virus challenge since vaccinated pigs showed clinical pseudorabies-specific syndromes. The deficiency in protection may due to the generation of late and low levels of gB antibodies and virus neutralizing antibodies.

关键词： pseudorabies virus RR1 and RR2 ribonucleotide reductase vaccine candidate

Enhancing transboundary natural tourism resources governance:unveiling the spatial pattern and its influencing factors

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Journal of Mountain Science 2024年第3期21卷 973-986页

作者： ZHANG Shengrui ZHANG tongyan JU Hongrun wang Yingjie Management College Ocean University of ChinaQingdao 266100China School of Tourism and Geography Science Qingdao UniversityQingdao 266071China Institute of Geographic Sciences and Natural Resources Research Chinese Academy of SciencesBeijing 100101China

Tourism resources that span provincial boundaries in China play a pivotal role in regional development,yet effective governance poses persistent challenges.This study addresses this issue by constructing a comprehensive database of transboundary natural tourism resources(TNTR)through amalgamation of diverse data sources.Utilizing the Getis-Ord Gi^(*),kernel density estimation,and geographical detectors,we scrutinize the spatial patterns of TNTR,focusing on both named and unnamed entities,while exploring the influencing factors.Our findings reveal 7883 identified TNTR in China,with mountain tourism resources emerging as the predominant type.Among provinces,Hunan boasts the highest count,while Shanghai exhibits the lowest.Southern China demonstrates a pronounced clustering trend in TNTR distribution,with the spatial arrangement of biological landscapes appearing more random compared to geological and water landscapes.Western China,characterized by intricate terrain,exhibits fewer TNTR,concurrently unveiling a significant presence of unnamed natural tourism resources.Crucially,administrative segmentation influences TNTR development,generating disparities in regional goals,developmental stages and intensities,and management approaches.In response to these variations,we advocate for strengthening the naming of the unnamed transboundary tourism resources,constructing a geographic database of TNTR for government and establishing a collaborative management mechanism based on TNTR database.Our research contributes to elucidating the intricate landscape of TNTR,offering insights for tailored governance strategies in the realm of cross-provincial tourism resource management.

关键词： Transboundary natural tourism resources(TNTR) Spatial difference Spatial autocorrelation Governance optimization China

非洲猪瘟病毒CP312R蛋白的真核表达及单克隆抗体制备

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中国兽医学报 2024年第3期44卷 445-449,457页

作者：颜世君赵少若郝丽影白露露王同燕邓均华田克恭国家兽用药品工程技术研究中心河南洛阳471000 普莱柯生物工程股份有限公司河南洛阳471000 洛阳普泰生物技术有限公司河南洛阳471000

为制备非洲猪瘟病毒(African swine fever virus,ASFV)CP312R蛋白的单克隆抗体,以真核表达ASFV的CP312R蛋白为免疫原免疫BALB/c小鼠制备单克隆抗体,并通过间接免疫荧光法(IFA)对获得的单克隆抗体进行反应性鉴定。结果利用杆状病毒表达系... 详细信息

为制备非洲猪瘟病毒(African swine fever virus,ASFV)CP312R蛋白的单克隆抗体,以真核表达ASFV的CP312R蛋白为免疫原免疫BALB/c小鼠制备单克隆抗体,并通过间接免疫荧光法(IFA)对获得的单克隆抗体进行反应性鉴定。结果利用杆状病毒表达系统(baculovirus expression system,BES)构建获得重组转移载体pOET3-CP312R,与杆状病毒基因组flashBAC^(TM)ULTRA共转染Sf9昆虫细胞获得重组杆状病毒AcMNPV-CP312R,且该病毒以胞内可溶形式表达出重组ASFV CP312R蛋白质。通过蛋白质印迹法鉴定显示重组CP312R蛋白可以与ASFV阳性血清发生特异性反应。此外,筛选获得2株针对重组CP312R蛋白的单克隆抗体(McAb),间接ELISA方法鉴定抗体滴度不低于1∶1024000。IFA试验检测表明,单克隆抗体与非洲猪瘟抗原发生特异性反应。本研究为非洲猪瘟亚单位疫苗研发及血清学检测方法的建立提供物质储备。

关键词：非洲猪瘟 CP312R蛋白重组杆状病毒单克隆抗体间接ELISA

表达猪瘟病毒E2蛋白的重组杆状病毒反应器工艺研究

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中国动物传染病学报 2024年第2期32卷 49-57页

作者：陈晓李伟国宋欢欢苏晓蕊王同燕谭菲菲田克恭国家兽用药品工程技术研究中心洛阳471000 普莱柯生物工程股份有限公司洛阳471000

为建立一套表达猪瘟病毒E2蛋白的重组杆状病毒反应器工艺,提高反应器上猪瘟病毒E2蛋白的表达量,以商业化培养基为基础,通过对关键营养组分添加量的DOE试验设计优化,获得一种组合补料配方,并在3 L反应器上对补料方式进行优化,以最大限度... 详细信息

为建立一套表达猪瘟病毒E2蛋白的重组杆状病毒反应器工艺,提高反应器上猪瘟病毒E2蛋白的表达量,以商业化培养基为基础,通过对关键营养组分添加量的DOE试验设计优化,获得一种组合补料配方,并在3 L反应器上对补料方式进行优化,以最大限度提高细胞密度。进一步在反应器上摸索了通气策略,以精确调控猪瘟E2重组杆状病毒培养过程的营养代谢平衡,解决副产物乳酸和氨过度积累导致细胞活力下降的问题。首次使用半连续补料工艺与渗透压动态平衡相结合的补料策略以及定向调节通气的工艺模式,使猪瘟病毒E2蛋白的表达量由130 mg/L增加至520 mg/L,提高了3倍。将3 L反应器工艺在100 L反应器上进行了连续3个批次的验证,结果猪瘟病毒E2蛋白的表达量均大于500 mg/L,收获猪瘟E2蛋白制备的疫苗抗体均在2周后转阳,且抗体水平持续上升,接种后8周抗体阻断率仍维持在80%左右。综上,3 L反应器工艺能够稳定放大至100 L反应器,建立了表达猪瘟病毒E2蛋白的重组杆状病毒100 L反应器工艺,且100 L反应器上重组蛋白的表达量稳定、免疫原性良好。

关键词：猪瘟病毒 E2蛋白重组杆状病毒关键营养单元半连续补料定向调节

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基因缺失杆状病毒载体的构建及应用

中国动物传染病学报 2024年第1期32卷 48-55页

作者：王同燕仝晓丹苏晓蕊宋欢欢李伟国刘武杰谭菲菲田克恭国家兽用药品工程技术研究中心洛阳471003 普莱柯生物工程股份有限公司洛阳471000 洛阳普泰生物技术有限公司洛阳471003

为解决杆状病毒表达系统存在的表达量低和毒种种子批代次窄问题,加快杆状病毒表达系统产业化进程。本研究首次通过Red和Cas9两种重组技术,先后对影响蛋白表达的V-cath、ChiA、P10和重组杆状病毒基因组稳定性的FP25K和DA26基因进行缺失... 详细信息

为解决杆状病毒表达系统存在的表达量低和毒种种子批代次窄问题,加快杆状病毒表达系统产业化进程。本研究首次通过Red和Cas9两种重组技术,先后对影响蛋白表达的V-cath、ChiA、P10和重组杆状病毒基因组稳定性的FP25K和DA26基因进行缺失。同时基于对CSFV-E2蛋白的结构和糖基化预测分析,突变影响同源二聚体二硫键形成的糖基化位点。最后通过悬浮转染的方式提高拯救病毒的滴度。结果表明对E2基因突变后,表达的E2蛋白95%以同源二聚体形式存在。通过供体质粒优化和病毒载体基因缺失,E2蛋白的表达水平提高约4倍,可稳定表达蛋白的重组杆状病毒毒株代次从P7代延长至P20代。悬浮转染获得的重组杆状病毒滴度提高约70倍,同时获得足量低代次的毒种,有效缩短从毒种构建到蛋白表达时间,有效解决杆状病毒表达系统表达量低和毒种种子批代次窄的问题,为猪瘟亚单位疫苗研发奠定基础。

关键词：杆状病毒猪瘟病毒基因缺失 E2蛋白

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Drogue detection for autonomous aerial refueling via hybrid pigeon-inspired optimized color opponent and saliency aggregation

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Chinese Journal of Aeronautics 2024年第5期37卷 27-38页

作者： tongyan WU Haibin DUAN Yanming FAN State Key Laboratory of Virtual Reality Technology and Systems School of Automation Science and Electrical EngineeringBeihang UniversityBeijing 100083China Virtual Reality Fundamental Research Laboratory Department of Mathematics and TheoriesPeng Cheng LaboratoryShenzhen 518000China AVIC Shenyang Aircraft Design and Research Institute Shenyang 110035China

Drogue detection is one of the challenging tasks in autonomous aerial refueling due to the requirement for accuracy and rapidity.Saliency detection based on image intrinsic cues can achieve fast detection,but with poor accuracy.Recent studies reveal that optimization-based methods provide accurate and quick solutions for saliency detection.This paper presents a hybrid pigeon-inspired optimization method,the optimized color opponent,that aims to adjust the weight of color opponent channels to detect the drogue region.It can optimize the weights in the selected aerial refueling scene offline,and the results are applied for drogue detection in the scene.A novel algorithm aggregated by the optimized color opponent and robust background detection is presented to provide better precision and robustness.Experimental results on benchmark datasets and aerial refueling images show that the proposed method successfully extracts the saliency region or drogue and exhibits superior performance against the other saliency detection methods with intrinsic cues.The algorithm designed in this paper is competent for the drogue detection task of autonomous aerial refueling.

关键词： Autonomous aerial refueling Drones Hybrid pigeon-inspired optimization Color opponent Saliency detection Saliency aggregation

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学科实践:义务教育语文课程的育人路径

课程．教材．教法 2022年第11期42卷 14-20页

作者：徐鹏王彤彦东北师范大学文学院长春130024 北京教育科学研究院基础教育教学研究中心北京100036

学科实践是促进学生在真实情境中理解、运用与转化学科知识的一种学科学习方式。《义务教育语文课程标准(2022年版)》倡导的学科实践以理解本位的学科知识观、整合取向的学科学习观为理论支点,以语文学习任务群为内容载体,以建构实践型... 详细信息

学科实践是促进学生在真实情境中理解、运用与转化学科知识的一种学科学习方式。《义务教育语文课程标准(2022年版)》倡导的学科实践以理解本位的学科知识观、整合取向的学科学习观为理论支点,以语文学习任务群为内容载体,以建构实践型育人路径为功能指向,体现了核心素养时代语境下语文学习方式变革的新方向。义务教育语文课程以学科实践为抓手,能够建构出具有语文学科特色的实践型育人路径。

关键词：学科实践语文课程学习任务群学科育人

A novel long persistent luminescent material CaGeO3:Tb3+ with photo-stimulation properties

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Journal of Rare Earths 2024年第7期42卷 1233-1239页

作者： Wei Meng Xiaoyan Fu tongyan Lin Zewen Liu Naihui Chen Jiaxu Zhang Runyao Liu Rujun Yang Long Chen Hongwu Zhang School of Chemistry and Material Sciences Ludong University College of Materials Science and Engineering Xiamen University of Technology College of Materials Xiamen University

A novel green long afterglow material CaGeO3:Tb3+was synthesized by high temperature solid phase method.According to the experimental results,CaGeO3:Tb3+is a green long persistent luminescent（LPL）material with excellent performance.CaGeO3:Tb3+shows LPL emission peak at 442,490.9,548.7,588.5and 623.8 nm,corresponding to5D3to7F4and5D4to7FJ（J=6,5,4,3） transitions of Tb3+,Its CIE coordinates are（0.27,0.50）,indicating that the afterglow emission is close to the light of green region.In addition,after the excitation source is turned off,the afterglow time is longer than 90 min.After an interval of 7 h,the photo-stimulation luminescence（PSL） can be observed by using 980 nm laser cyclic excitation.The thermoluminescence（ThL） results show that there are three types of traps in the material and their depths are 0.666,0.774 and 0.892 eV respectively,which are caused by the doping of Tb3+.All the results show that CaGeO3:Tb3+is an excellent long afterglow luminescent material.

关键词： CaGeO3:Tb3+ Long persistent luminescence Photo-stimulation Rare earths Trap