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Yansheng Geng, Chenyan Zhao, Weijin Huang, Xuanpu Wang, Ying Xu, Dongxue Wu, Yueliang Du, Huan Liu, Youchun Wang
Hepatitis E virus (HEV) is an important human pathogen, with pigs and other species serving as natural animal reservoirs. Recently, the report of detection of genotype 4 HEV in dairy milk with high positive rate in Yunnan province of China has attracted extensive attention. To assess the zoonotic risk of cows as HEV reservoir and transmission of HEV through dairy milk, 467 fecal samples of cows, 276 fresh milk samples, and 140 retail milk samples were collected across Hebei Province, China, from March 2017 to May 2018, and detected for HEV RNA...
November 6, 2018: International Journal of Food Microbiology
Donglai Liu, Xinyuan Zhang, Haiwei Zhou, Xiaojing Lin, Dawei Shi, Shu Shen, Yabin Tian, Bo Du, Henghui Zhang, Haibo Wang, Youchun Wang, Chuntao Zhang
Background: Liquid biopsies based on next-generation sequencing (NGS) assays are confronted with more opportunities and challenges. Widespread clinical implementation of NGS-based cancer in vitro diagnostic tests (IVDs) highlighted the urgency to establish reference materials (RMs) which could provide full control of the process from nucleic acid extraction to test report generation. Quality control based on cell-free DNA (cfDNA) RMs is especially important for liquid biopsies. Methods: Here, we used genomic DNA from thirteen cell lines to establish four negative cfDNA RMs (N1-N4) and four multiplex cfDNA RMs (L1-L4) at serial allelic frequencies ranging from approximately 2% to 0...
2018: Journal of Cancer
Changfa Fan, Xi Wu, Qiang Liu, Qianqian Li, Susu Liu, Jianjun Lu, Yanwei Yang, Yuan Cao, Weijin Huang, Chunnan Liang, Tianlei Ying, Shibo Jiang, Youchun Wang
Infection by the Middle East respiratory syndrome coronavirus (MERS-CoV) causes respiratory illness and has a high mortality rate (~35%). The requirement for the virus to be manipulated in a biosafety level three (BSL-3) facility has impeded development of urgently-needed antiviral agents. Here, we established anovel mouse model by inserting human dipeptidyl peptidase 4 (hDPP4) into the Rosa26 locus using CRISPR/Cas9, resulting in global expression of the transgene in a genetically stable mouse line. The mice were highly susceptible to infection by MERS-CoV clinical strain hCoV-EMC, which induced severe diffuse pulmonary disease in the animals, and could also be infected by an optimized pseudotyped MERS-CoV...
August 23, 2018: Viruses
Xin Zhang, Qiang Liu, Qianqian Li, Yinghong Li, Zhandong Liu, Hongbin Deng, Sheng Tang, Yanxiang Wang, Youchun Wang, Danqing Song
Twenty-six novel tricyclic sophoridinic and matrinic derivatives containing a common chlorinated benzene fragment were designed, synthesized and evaluated for their anti-ebolavirus (EBOV) activities. Structure-activity relationship analysis indicated: (i) 12 N -dichlorobenzyl motif was beneficial for the activity; (ii) the chiral configuration at C5 atom might not affect the activity much. Among the target compounds, compound 7d exhibited the most potent potency against EBOV with an IC50 value of 5.29 ╬╝mol/L and an SI value of over 37...
July 2018: Acta Pharmaceutica Sinica. B
Junge Chen, Youchun Xu, He Yan, Yunzeng Zhu, Lei Wang, Yan Zhang, Ying Lu, Wanli Xing
Bacterial infections may cause severe diseases such as tuberculosis, sepsis, nephritis and cystitis. The rapid and sensitive detection of bacteria is a prerequisite for the treatment of these diseases. The current gold standard for bacterial identification is bacteriological culture. However, culture-based identification takes 3-7 days, which is time-consuming and laborious. In this study, bacteria in urine samples were enriched using a portable filter-based pipette. Then, a centrifugal chip was constructed to detect multiple pathogenic bacteria from urine samples by integrating the DNA extraction, multiplex recombinase polymerase amplification (RPA) and fluorescent detection together...
August 7, 2018: Lab on a Chip
Tingting Ning, Jianhui Nie, Weijin Huang, Changgui Li, Xuguang Li, Qiang Liu, Hui Zhao, Youchun Wang
Background: Since its emergence in 2013, there have been five H7N9 influenza epidemic waves in China. However, evolution of the hemagglutinin (HA) antigenicity has not been systematically investigated. Methods: To better understand how antigenic drift in HA proteins of H7N9 occurs, 902 H7N9 HA sequences from public database were retrieved and analyzed. 53 mutants with single amino acid substitutions in HA were introduced into pseudoviruses and their antigenic characteristics were analyzed using pseudovirus-based assays...
July 5, 2018: Journal of Infectious Diseases
Jiaxiong Tan, Shaohua Chen, Jingying Huang, Youchun Chen, Lijian Yang, Chunli Wang, Jun Zhong, Yuhong Lu, Liang Wang, Kanger Zhu, Yangqiu Li
AIM: The immunosuppressive microenvironment plays a crucial role in T-cell immunodeficiency in multiple myeloma (MM). Overexpression of T-cell immunosuppressive receptors, including programmed death-1 (PD-1) and T-cell immunoglobulin and mucin-domain-containing-3 (Tim-3), may be related to tumor immunosuppression and poor prognosis, and the malignant bone marrow (BM) microenvironment may contribute to such immunosuppression. The purpose of this study was to analyze the distribution of PD-1+ and/or Tim-3+ T cells in different T-cell subset in patients with MM...
June 25, 2018: Asia-Pacific Journal of Clinical Oncology
Hui Xie, Jianhui Nie, Qingqing Chen, Weijin Huang, Youchun Wang
Although a number of standardized human immunodeficiency virus 1 (HIV-1) pseudoviruses have been generated to assess neutralizing antibodies, subtype B/B' has not been comprehensively characterized either genotypically or phenotypically. Full-length env genes were isolated from plasma samples derived from B/B'-infected individuals, including former plasma donors and men who had sex with men. The neutralization properties of these pseudoviruses were determined by testing against sCD4, 16 neutralizing monoclonal antibodies and 44 plasma samples, including samples from those infected with the three major prevalent clades in China...
May 16, 2018: Emerging Microbes & Infections
Donglai Liu, Haiwei Zhou, Dawei Shi, Shu Shen, Yabin Tian, Lin Wang, Jiatao Lou, Rong Cong, Juan Lu, Henghui Zhang, Meiru Zhao, Shida Zhu, Zhisheng Cao, Ruilin Jin, Yin Wang, Xiaoni Zhang, Guohua Yang, Youchun Wang, Chuntao Zhang
Background: Widespread clinical implementation of next-generation sequencing (NGS)-based cancer in vitro diagnostic tests (IVDs) highlighted the urgency to establish reference materials which could provide full control of the process from nucleic acid extraction to test report generation. The formalin-fixed, paraffin-embedded (FFPE) tissue and blood plasma containing circulating tumor deoxyribonucleic acid (ctDNA) were mostly used for clinically detecting onco-relevant mutations. Methods: We respectively developed multiplex FFPE and plasma reference materials covering three clinically onco-relevant mutations within the epidermal growth factor receptor ( EGFR ) gene at serial allelic frequencies...
2018: Journal of Cancer
Wenlong Lin, Ning Wang, Kangxing Zhou, Fasheng Su, Yu Jiang, Jianan Shou, Huan Liu, Chunmei Ma, Youchun Qian, Kai Wang, Xiaojian Wang
Th17 cells contribute to the development of autoimmune diseases by secreting interleukin-17 (IL-17), which activates its receptor (IL-17R) that is expressed on epithelial cells, macrophages, microglia, and resident neuroectodermal cells. However, the mechanisms through which IL-17R-mediated signaling contributes to the development of autoimmune disease have not been completely elucidated. Here, we demonstrate that Raf-1 kinase inhibitor protein (RKIP) deficiency in mice ameliorates the symptoms of experimental autoimmune encephalomyelitis (EAE)...
June 2018: EMBO Reports
Qianqian Li, Qiang Liu, Weijin Huang, Xuguang Li, Youchun Wang
Emerging and reemerging infectious diseases have a strong negative impact on public health. However, because many of these pathogens must be handled in biosafety level, 3 or 4 containment laboratories, research and development of antivirals or vaccines against these diseases are often impeded. Alternative approaches to address this issue have been vigorously pursued, particularly the use of pseudoviruses in place of wild-type viruses. As pseudoviruses have been deprived of certain gene sequences of the virulent virus, they can be handled in biosafety level 2 laboratories...
January 2018: Reviews in Medical Virology
Jiajing Wu, Chenyan Zhao, Qiang Liu, Weijin Huang, Youchun Wang
Chikungunya virus (CHIKV) is an arthropod-borne virus that is transmitted to humans primarily via the bite of an infected mosquito. Infection of humans by CHIKV can cause chikungunya fever which is an acute febrile illness associated with severe, often debilitating polyarthralgias. Since a re-emergence of CHIKV in 2004, the virus has spread into novel locations in nearly 40 countries including non-endemic regions and has led to millions of cases of disease throughout countries. Handling of CHIKV is restricted to the high-containment Biosafety Level 3 (BSL-3) facilities, which greatly impede the research progress of this virus...
November 7, 2017: Vaccine
Tingting Ning, Aaron Wolfe, Jianhui Nie, Weijin Huang, Xiaojiang S Chen, Youchun Wang
Background: Each vaccine for human papillomavirus type 16 (HPV16) has been developed on the basis of a single variant, and whether these vaccines can prevent infection due to naturally occurring variants was not clear. Methods: To examine this question, constructs of 39 naturally occurring single amino acid substitutions in L1 were generated for pseudovirion production, based on the analysis of 1204 HPV16 L1 protein sequences from the National Center for Biotechnology Information and Papilloma Virus Episteme...
October 17, 2017: Journal of Infectious Diseases
Qingqing Chen, Jianhui Nie, Weijin Huang, Yanmei Jiao, Lan Li, Tong Zhang, Juan Zhao, Hao Wu, Youchun Wang
Sensitive assays for HIV-1 neutralizing antibody detection are urgently needed for vaccine immunogen optimization and identification of protective immune response levels. In this study, we developed an easy-to-use HIV-1 pseudovirus neutralization assay based on a human CD4+ lymphoblastoid cell line A3R5 by employing a high efficient pseudovirus production system. Optimal conditions for cell counts, infection time, virus dose and concentration of DEAE-dextran were tested and identified. For T-cell line-adapted tier 1 virus strains, significantly higher inhibitory efficiency was observed for both monoclonal neutralizing antibody (4 fold) and plasma (2 fold) samples in A3R5 than those in TZM-bl assay...
January 2, 2018: Human Vaccines & Immunotherapeutics
Chenyan Zhao, Yansheng Geng, Weijing Huang, Hongxia Ma, Youchun Wang
Nucleotide-based methods are conventionally used to classify the hepatitis E virus (HEV) genotypes. A serological enzyme immunoassay (EIA) using open reading frame 3 (ORF3) C-terminal peptides was developed to conveniently and accurately classify and evaluate the genotypes of HEV. The sera of mice immunized with HEV genotype 1, 3, and 4 reacted highly specifically to the peptides of the corresponding genotypes. Most (84.2%) clinical sera infected with HEV genotype 4 were positive for anti-HEV antibodies when tested with the ORF3 peptides of genotype 4, but were negative for genotypes 1 and 3...
January 2018: Journal of Medical Virology
Xueling Wu, Jun Yang, Tao Na, Kehua Zhang, Andrew M Davidoff, Bao-Zhu Yuan, Youchun Wang
The stimulator of interferon genes (STING) protein has emerged as a critical signal transduction molecule in the innate immune response. Sustained activation of the STING signaling induced by cytosolic DNA has been considered to be the cause of a variety of autoimmune diseases characterized by uncontrolled inflammation. Therefore, it is important to understand the molecular basis of the regulation of STING signaling pathway. Here we demonstrate that the STING protein undergoes a proteasome-mediated degradation in human diploid cell (HDC) lines including MRC-5 following the transfection of double-stranded DNA (dsDNA)...
2017: PloS One
Qianqian Li, Qiang Liu, Weijin Huang, Jiajing Wu, Jianhui Nie, Meng Wang, Chenyan Zhao, Li Zhang, Youchun Wang
Lassa virus (LASV) causes a severe hemorrhagic fever endemic throughout western Africa. Because of the ability to cause lethal disease in humans, limited treatment options, and potential as a bioweapon, the need for vaccines to prevent LASV epidemic is urgent. However, LASV vaccine development has been hindered by the lack of appropriate small animal models for efficacy evaluation independent of biosafety level four (BSL-4) facilities. Here we generated an LASV-glycoprotein precursor (GPC)-pseudotyped Human immunodeficiency virus containing firefly luciferase (Fluc) reporter gene as surrogate to develop a bioluminescent-imaging-based BALB/c mouse model for one-round infection under non-BSL-4 conditions, in which the bioluminescent intensity of Fluc was utilized as endpoint when evaluating vaccine efficacy...
September 12, 2017: Vaccine
Yunzeng Zhu, Yiqi Chen, Xiangrui Meng, Jing Wang, Ying Lu, Youchun Xu, Jing Cheng
Centrifugal microfluidics has been widely applied in the sample-in-answer-out systems for the analyses of nucleic acids, proteins, and small molecules. However, the inherent characteristic of unidirectional fluid propulsion limits the flexibility of these fluidic chips. Providing an extra degree of freedom to allow the unconstrained and reversible pumping of liquid is an effective strategy to address this limitation. In this study, a wirelessly charged centrifugal microfluidic platform with two rotation axes has been constructed and the flow control strategy in such platform with two degrees of freedom was comprehensively studied for the first time...
September 5, 2017: Analytical Chemistry
Qiang Liu, Shuya Zhou, Changfa Fan, Weijin Huang, Qianqian Li, Susu Liu, Xi Wu, Baowen Li, Youchun Wang
As concerns increase about adenovirus type 5 (Ad5) being a safe gene transfer vector, it is important to evaluate its distribution, residence time, and possible toxicity in immunodeficient populations. To characterize the potential risk associated with different Ad5 vector delivery modes, we used immunocompetent and immunodeficient Rag2 (-/-) animals to establish mouse and rat models that could be monitored with bioluminescent imaging following intramuscular or intravascular infection with an engineered replication-incompetent Ad5 virus carrying the firefly luciferase gene (Ad5-Fluc)...
June 15, 2017: Scientific Reports
Li Zhang, Qianqian Li, Qiang Liu, Weijin Huang, Jianhui Nie, Youchun Wang
Marburg virus (MARV) can cause lethal hemorrhagic fever in humans. Handling of MARV is restricted to high-containment biosafety level 4 (BSL-4) facilities, which greatly impedes research into this virus. In this study, a high titer of MARV pseudovirus was generated through optimization of the HIV backbone vectors, the ratio of backbone vector to MARV glycoprotein expression vector, and the transfection reagents. An in vitro neutralization assay and an in vivo bioluminescent imaging mouse model for MARV were developed based on the pseudovirus...
August 3, 2017: Human Vaccines & Immunotherapeutics
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