Zengjun Lu

Chinese Academy of Agricultural Sciences

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Zaixin Liu

Lanzhou Veterinary Research Institute

Pu Sun

Chinese Academy of Agricultural Sciences

Pinghua Li

Lanzhou Veterinary Research Institute

Yimei Cao

Lanzhou Veterinary Research Institute

Xingwen Bai

Lanzhou Veterinary Research Institute

Huifang Bao

Lanzhou Veterinary Research Institute

Yuanfang Fu

Lanzhou Veterinary Research Institute

Kun Li

Chinese Academy of Agricultural Sciences

Yingli Chen

Development Center for Biotechnology

Xueqing Ma

Chinese Academy of Agricultural Sciences

Cooperative Institutions

Chinese Academy of Agricultural Sciences

Lanzhou Veterinary Research Institute

Lanzhou University

Chinese Academy of Sciences

Gansu Provincial Center for Disease Control and Prevention

Gansu Agricultural University

Ministry of Agriculture and Rural Affairs

University of Chinese Academy of Sciences

Sichuan University

Huazhong Agricultural University

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[Development of a sandwich ELISA for detecting 3AB non-structural protein of foot-and-mouth disease virus].

Chinese journal of biotechnology/Shengwu gongcheng xuebao (2020)

Yuanfang Fu Wei He Pu Sun Lin Yang Huifang Bao

10.13345/j.cjb.200149

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Research Progress on Species Identification of Sarcosaprophagous Flies.

PubMed (2021)

Yujie Ran X Y Zhang Zengjun Lu

Estimation of postmortem interval （PMI） has always been one of the difficult problems for forensic scientists. It is especially hard to estimate the PMI of highly decomposed corpses in the wild or in secluded houses with conventional methods. Therefore, application of insect evidence at the scene is usually required for estimation. Sarcosaprophagous flies of different species have totally different developmental rates. In actual cases, direct measurement of the body length of the larvae, calculation of accumulated temperature and succession stages without species identification, or calculation based on incorrect species identification would often lead to a large deviation between the calculated results and the real PMI. This mistake would also mislead the case investigation. Therefore, accurate species identification should be implemented before any PMI estimation of decomposed corpses with forensic entomological methods. This article reviews the general and ultramicroscopic species identification and molecular biological species identification methods of different stages of sarcosaprophagous flies, in order to provide new ideas and methods for related research and practice, and provide reference for the application and promotion of forensic entomology in the front line of public security.嗜尸性蝇类种属鉴定研究进展.死亡时间（postmortem interval，PMI）推断一直是困扰法医学工作者的难题之一，尤其是荒郊野外或偏僻屋内的高度腐败尸体更是难以使用常规手段推断PMI，因此常需要借助现场中的昆虫证据来推断。不同种属的蝇类发育时间差异较大，在实际案件中，如果不进行种属鉴定而直接测量蝇蛆体长、计算积温或演替阶段，或是基于错误的种属鉴定来计算，往往会导致推算出的结果与实际PMI之间出现较大偏差，误导案件侦查方向。因此，使用法医昆虫学方法来推断腐败尸体PMI，必须先进行准确的昆虫种属鉴定。本文对嗜尸性蝇类不同虫态的大体和超微形态学种属鉴定以及分子生物学种属鉴定方法进行综述，以期为相关研究和实践提供新思路和新方法，为法医昆虫学在鉴定实践的应用推广提供参考。.法医昆虫学；嗜尸性蝇类；种属鉴定；综述.

Forensic Entomology

Identification

Mistake

Species Identification

Forensic Genetics

10.12116/j.issn.1004-5619.2021.410208

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[Expression of recombinant plasmid pcDNA3.1/P12X3C with multi-genes of foot-and-mouth disease virus in BHK-21 cells].

PubMed (2003)

Huichen Guo Zaixin Liu Shiqi Sun Zengjun Lu Guangqing Zhou

In order to obtain the gene P12X3C of Foot-and-Mouth Disease Virus (FMDV) that includes full length P1, 2A, 3C and a part of 2B, the site mutation strategy was used. After being digested by Kpn I and Xba I respectively, the gene P12X3C was cloned into the pcDNA3.1 (+) expression vector. The recombinant plasmid was checked by restriction enzyme analysis and nucleic acid sequencing, and then named pcDNA3.1/P12X3C. Further, BHK-21 cells was transfected with pcDNA3.1/P12X3C by using lipoid. The proteins of Foot-and-Mouth Disease Virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy. The result shows the gene P12X3C is cloned into eukaryotic expression plasmid, and the recombinant eukaryotic expression plasmid pcDNA3.1/P12X3C could express proteins of Foot-and-Mouth Disease Virus in BHK-21 cells, which have immunocompetence. This study demonstrates that delivery of a recombinant eukaryotic expression plasmid containing P12X3C coding regions results in the assembly of FMDV capsid structures, which will offer experimental base to DNA vaccine of FMDV.

Recombinant virus

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Porcine antibodies reveal novel non-neutralizing universal epitopes on FMDV and their overlaps with neutralization sites

Veterinary Microbiology (2025)

Ying Sun Shenglin Huang Fengjuan Li Shulun Huang Pinghua Li

10.1016/j.vetmic.2025.110440

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[Diversity of receptor recognition site of type Asial foot-and-mouth disease virus]

Pinghua Li Xingwen Bai Pu Sun Zengjun Lu Weijun Cao

Objective Foot-and-Mouth Disease Virus (FMDV) initiates infection by binding to integrin receptors via a highly conserved Arg-Gly-Asp (RGD) sequence found in the G-H loop of the structural protein VP1. However, FMDV is an RNA virus, which harbors the evolutionary potential to render the RGD motif dispensable upon changes in constant environment. We studied mutation of RGD motif upon short-time passages FMDV Asial/JS/China/2005 field strain in different host. Methods VP1 gene was amplified from Asial/JS/China/05 field strain, the fourth passage virus in sulking mice (MF4) of the above strain and the virus isolated from a pig housing with cattle inoculated with the above strain followed eight passages in BHK-21 cell (PBF8) by RT-PCR, and the VP1 genes were sequenced and their deduced amino acid sequences were compared with each other. Result Dominant population with RGD and Arg-Ser-Asp (RSD) receptor recognition site motif was generated after four passages of Asial/JS/China/2005 field virus in sulking mice and another dominant population with Arg-Asp-Asp (RDD) motif was produced after eight passages the virus isolated from housing pig with cattle inoculated with the above field strain. Conclusion This study indicated that the dominant FMDV virus populations with RSD or RDD receptor binding site instead of original RGD motif were produced upon short-time evolution of FMDV field isolates with RGD motif in different environment. These studies not only increase number of viable mutants with substitutions in the RGD region, but also these profoundly altered, but viable, mutants with different receptor recognition site will provide useful tools for studies of cell recognition by FMDV and host tropism modifications.

RGD motif

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Molecular vaccine prepared by fusion of XCL1 to the multi-epitope protein of foot-and-mouth disease virus enhances the specific humoural immune response in cattle

Applied Microbiology and Biotechnology (2017)

Kun Li Huifang Bao Guoyan Wei Dong Li Yingli Chen

10.1007/s00253-017-8523-y

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[Rescue and identification foot-and-mouth disease virus Asia1/JS/China/2005 strain with Arg-Gly-Asp RGD receptor recognition site].

PubMed (2009)

Pinghua Li Xingwen Bai Weijun Cao Zengjun Lu Pu Sun

To construct an infectious full-length cDNA clone of Asial/JS/China/2005 strain with Arg-Gly-Asp (RGD) receptor recognition site.We constructed foot-and-mouth disease virus type Asial full-length cDNA clone pFMDV-RGD by using site-directed mutagenesis. The plasmid pFMDV-RGD contained the desired mutation. The plasmids pFMDV-RGD were linearized with NotI enzyme. Linearized plasmid and pcDNAT7P plasmids expressing T7 RNA polymerase cotransfected into BHK-21 cells to rescue FMDV-RGD.We constructed FMDV Asial/JS/China/2005 strain full-length cDNA clone with Arg-Gly-Asp receptor recognition site by sequence. We obtained rescued virus by plasmid cotransfection. The results of sequencing, indirect immunofluorescence, electron microscope and sulk mice pathogenicity analysis showed foot-and-mouth disease virus containing Arg-Gly-Asp receptor recognition site was successfully rescued.The results lay a foundation for further study of biology characteristic diversity of rescued virus with Arg-Gly-Asp and Arg-Asp-Asp (RDD) receptor recognition site.

clone (Java method)

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[Secreted expression of nonstructural protein gene 3ABC of foot-and-mouth disease virus in Sf9 cells and activity analysis].

Jiangtao Ma Zengjun Lu Yimei Cao Guo Huichen Guo Jian-hong

Entire 3ABC sequence of FMDV containing a 6×his tag coding sequence at the N-terminal was obtained through PCR amplification using a pair of specific primers, subcloned into shuttle plasmid of pMelBac-B with a melittin secretion signal sequence and finally constructed recombinant plasmid of pMel-3ABC. After co-transfected the recombinant plasmid and linearized Bac-N-Blue TM DNA into Sf9 insect cell under intermediary agent of the Cellfectin+, the result showed that we have already acquired recombinant baculovirus by screen of plaque assay and identification of PCR. Though the recombinant baculovirus infecting the Sf9 cells again, experiments indicated that 3ABC gene could express in insect cells and the expressed protein was secreted in the supernatant of Sf9 cell culture possessing favourable biological activities detected by adopting two methods of SDS-PAGE and Western blot. The result verified that the protein could respond with sera derived from FMDV infected animals, but have no responsibility with sera derived from health animals and vaccinated animals detected by indirect ELISA using antigen of expressed protein after purification with Ni-NTA his bind resin. Therefore, this study has established a solid foundation for establishing an effective diagnosis method to discriminating the FMDV infected animals from vaccinated animals.

Sf9

Recombinant virus

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[Transcriptomic differences between the spleens of mice immunized with inactivated antigens of foot-and-mouth disease virus and Senecavirus A].

PubMed (2024)

Zixuan Zheng Xueqing Ma Kun Li Pu Sun Shulun Huang

The aim of this study was to compare the immune responses of C57BL/6 mice immunized with two pathogens, foot-and-mouth disease virus (FMDV) and Senecavirus A (SVA), and to provide clues for revealing the regulatory mechanisms of acquired immunity. Inactivated and purified FMDV and SVA antigens were used to immunize C57BL/6 mice respectively, and the mice immunized with PBS were taken as the control. The percentages of Th1 and Th2 cells in the spleen lymphocytes of mice in each group were analyzed by flow cytometry at 14 and 28 days after immunization. RNA-Seq was performed for the spleen. Mouse macrophages were stimulated with the antigens

Inactivated vaccine

10.13345/j.cjb.240084

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A multiplex RT-PCR for simultaneous differentiation of three viral pathogens of penaeid shrimp

Diseases of Aquatic Organisms (2007)

Zhiqin Xie Ying Pang Xianwen Deng Xiaofei Tang Jinfeng Liu

A multiplex reverse transcription polymerase chain reaction (mRT-PCR) was developed and optimized to simultaneously detect 3 viral pathogens of shrimp. Three sets of specific oligonucleotide primers for Taura syndrome virus (TSV), white spot syndrome virus (WSSV) and infectious hypodermal and hematopoietic necrosis virus (IHHNV) were used in the assay. The mRT-PCR DNA products were visualized by gel electrophoresis and consisted of fragments of 231 bp for TSV, 593 bp for WSSV and 356 bp for IHHNV. No specific bands of the same size were amplified from other penaeid shrimp pathogenic viruses or bacteria. As little as 10 pg of TSV RNA and 100 pg of WSSV DNA and IHHNV DNA could be detected using gel electrophoresis. Studies are in progress to further test the specificity and sensitivity of this mRT-PCR method on viral isolates, as well as on clinical samples.

White spot syndrome

Multiplex

DNA virus

10.3354/dao076077

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Citations (14)