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Identification of fangchinoline as a broad-spectrum enterovirus inhibitor through reporter virus based high-content screening

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Virologica Sinica 2024年第2期39卷 301-308页

作者： Qiu-Yan Zhang Jia-Qi Li Qi Li Yang Zhang Zhe-Rui Zhang Xiao-Dan Li Hong-Qing Zhang Cheng-Lin Deng Feng-Xia Yang Yi Xu Bo Zhang The Joint Center of Translational Precision Medicine Department of Infections and DiseasesGuangzhou Institute of PediatricsGuangzhou Women and Children's Medical CenterGuangzhou510623China Key Laboratory of Special Pathogens and Biosafety Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan430071China University of Science and Technology of China Department of Life Sciences and MedicineHefei230026China Hunan Normal University School of MedicineChangsha410081China

Hand,foot,and mouth disease(HFMD)is a common pediatric illness mainly caused by enteroviruses,which are important human ***,there are no available antiviral agents for the therapy of enterovirus *** this study,an excellent high-content antiviral screening system utilizing the EV-A71-eGFP reporter virus was *** this screening system,we screened a drug library containing 1042 natural compounds to identify potential EV-A71 ***(FAN),a bis-benzylisoquinoline alkaloid,exhibits potential inhibitory effects against various enteroviruses that cause HFMD,such as EV-A71,CV-A10,CV-B3 and *** investigations revealed that FAN targets the early stage of the enterovirus life *** the selection of FAN-resistant EV-A71 viruses,we demonstrated that the VP1 protein could be a potential target of FAN,as two mutations in VP1(E145G and V258I)resulted in viral resistance to *** research suggests that FAN is an efficient inhibitor of EV-A71 and has the potential to be a broad-spectrum antiviral drug against human enteroviruses.

关键词： Hand,foot,and mouth disease(HFMD) enterovirus EV-A71 High-content screening Fangchinoline(FAN)

Integration of HiBiT into enteroviruses:A universal tool for advancing enterovirus virology research

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Virologica Sinica 2024年第3期39卷 422-433页

作者： Rui Yu Xiaohong Li Peng Zhang Minghao Xu Jitong Zhao Jingjing Yan Chenli Qiu Jiayi Shu Shuo Zhang Miaomiao Kang Xiaoyan Zhang Jianqing Xu Shuye Zhang School of Medicine Shanghai UniversityShanghai200444China Clinical Center for Biotherapy Zhongshan HospitalFudan UniversityShanghai200433China Guangzhou Institutes of Biomedicine and Health The Chinese Academy of SciencesGuangzhou510530China Institutes of Biomedical Sciences Fudan UniversityShanghai200032China Shanghai Geriatric Medical Center Shanghai201104China Clinical Center for Biotherapy Zhongshan Hospital/Zhongshan Hospital(Xiamen)Fudan University361015China Shanghai Public Health Clinical Center Fudan UniversityShanghai201508China

The utilization of enteroviruses engineered with reporter genes serves as a valuable tool for advancing our understanding of enterovirus biology and its applications,enabling the development of effective therapeutic and preventive *** this study,our initial attempts to introduce a NanoLuc luciferase(NLuc)reporter gene into recombinant enteroviruses were unsuccessful in rescuing viable *** hypothesized that the size of the inserted tag might be a determining factor in the rescue of the ***,we inserted the 11-amino-acid HiBiT tag into the genomes of enterovirus A71(EV-A71),coxsackievirus A10(CVA10),coxsackievirus A7(CVA7),coxsackievirus A16(CVA16),namely EV-A71-HiBiT,CVA16-HiBiT,CVA10-HiBiT,CVA7-HiBiT,and observed that the HiBiT-tagged viruses exhibited remarkably high rescue ***,the HiBiT-tagged enteroviruses displayed comparable characteristics to the wild-type viruses.A direct comparison between CVA16-NLuc and CVA16-HiBiT recombinant viruses revealed that the tiny HiBiT insertion had minimal impact on virus infectivity and replication ***,these HiBiT-tagged enteroviruses demonstrated high genetic stability in different cell lines over multiple *** addition,the HiBiT-tagged viruses were successfully tested in antiviral drug assays,and the sensitivity of the viruses to drugs was not affected by the HiBiT ***,our findings provide definitive evidence that the integration of HiBiT into enteroviruses presents a universal,convenient,and invaluable method for advancing research in the realm of enterovirus ***,HiBiT-tagged enteroviruses exhibit great potential for diverse applications,including the development of antivirals and the elucidation of viral infection mechanisms.

关键词： enterovirus Infectious clones Reporter genes Luciferase Drug screening

Mitochondria Redistribution in enterovirus A71 Infected Cells and Its Effect on Virus Replication

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Virologica Sinica 2019年第4期34卷 397-411页

作者： Yang Yang Haolong Cong Ning Du Xiaodong Han Lei Song Wenliang Zhang Chunrui Li Po Tien Center for Molecular Virology CAS Key Laboratory of Pathogenic Microbiology and ImmunologyInstitute of MicrobiologyChinese Academy of SciencesBeijing 100101China University of the Chinese Academy of Sciences Beijing 100101China Beijing Institutes of Life Science Chinese Academy of SciencesBeijing 100101China College of Life Sciences Inner Mongolia Agriculture UniversityHohhot 010018China

enterovirus A71(EV-A71)is one of the main causative agents of hand,foot and mouth disease(HFMD)and it also causes severe neurologic complications in infected *** interactions between some viruses and the host mitochondria are crucial for virus replication and *** this study,it was observed that EV-A71 infection resulted in a perinuclear redistribution of the *** mitochondria rearrangement was found to require the microtubule network,the dynein complex and a low cytosolic calcium ***,the EV-A71 non-structural protein 2BC was identified as the viral protein capable of inducing mitochondria *** protein was found localized on mitochondria and interacted with the mitochondrial Rho GTPase 1(RHOT1)that is a key protein required for attachment between the mitochondria and the motor proteins,which are responsible for the control of mitochondria ***,suppressing mitochondria clustering by treating cells with nocodazole,EHNA,thapsigargin or A23187 consistently inhibited EV?A71 replication,indicating that mitochondria recruitment played a crucial role in the EV-A71 life *** study identified a novel function of the EV-A71 2BC protein and provided a potential model for the regulation of mitochondrial motility in EV-A71 infection.

关键词： enterovirus A71(EV-A71) Mitochondria Microtubule network Calcium concentration Mitochondrial Rho GTPase 1(RHOT1)

Coxsackievirus A6 was the most common enterovirus serotype causing hand,foot,and mouth disease in Shiyan City,central China

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World Journal of Clinical Cases 2022年第31期10卷 11358-11370页

作者： Jing-Feng Li Chuan-Jie Zhang Ya-Wei Li Chao Li Shi-Chao Zhang Sha-Sha Wang Yong Jiang Xin-Bing Luo Xing-Juan Liao Shou-Xin Wu Ling Lin Department of Pediatrics Taihe HospitalAffiliated Hospital of Hubei University of MedicineShiyan 442000Hubei ProvinceChina Department of Children Health Care Wuhan Children's Hospital(Wuhan Maternal and Child Healthcare Hospital)Tongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430061Hubei ProvinceChina Department of Health Services Taihe HospitalAffiliated Hospital of Hubei University of MedicineShiyan 442000Hubei ProvinceChina Department of Pharmaceuticals Shanghai Biotecan Pharmaceuticals Co.Ltd.Shanghai 200000China Zhangjiang Center for Translational Medicine Shanghai Zhangjiang Institute of Medical InnovationShanghai 442000China

BACKGROUND Hand,foot,and mouth disease(HFMD)has become one of the most common infectious diseases in *** 2016,the primary causal serotypes were enterovirus A71(EV-A71)and coxsackievirus A16(CV-A16).Following the introduction of EV-A71 vaccines in China since 2016,the situation could ***-A6 has recently replaced EV-A71 and CV-A16 in some areas of ***,the epidemiological characteristics of central China remain *** To investigate the clinical symptoms and pathogen spectrum of HFMD in Shiyan City,central China,in recent *** The epidemiological,clinical,and laboratory data from HFMD cases reported to the Shiyan Center for Disease Control and Prevention between January 2016 and December 2020 were analyzed.196 throat swab specimens were collected from hospitalized HFMD patients between January 2018 and December *** detect and genotype enteroviruses,real-time reverse transcription-polymerase chain reaction and sequencing of the 5'-untranslated region were *** Shiyan,168 laboratory-confirmed HFMD cases were studied using a logistic regression model to determine the effect of predominant enterovirus *** on the logistic regression model,the least absolute shrinkage and selection operator model was used to analyze the correlation between CV-A6 infection and various clinical characteristics in HFMD patients in *** From 2016 to 2020,35840 HFMD cases were reported in *** number of cases decreased by 48.4%from 2016 to *** 1.58-fold increases were found in 2018 and 2019 when compared to the previous year,*** 2020,a decrease of about 85.5%was reported when compared to *** most common serotypes shifted from EV-A71 and CV-A16(about 60%-80%in 2016 and 2018)to others(more than 80.0%in 2017,2019,and 2020).EV-A71 lost its dominance in 2017 in *** 196 confirmed HFMD cases,85.7%tested positive for enterovirus,with CV-A6 being the most common serotype(121/168,72.0%).The po

关键词： Hand,foot,and mouth disease Coxsackievirus A6 enterovirus Prevalence serotype Fever Myocardial damage

Clinical and neuroimaging features of enterovirus71 related acute flaccid paralysis in patients with hand-foot-mouth disease

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Asian Pacific Journal of Tropical Medicine 2013年第1期6卷 68-72页

作者： Feng Chen Jian-Jun Li Tao Liu Guo-Qiang Wen Wei Xiang Department of Radiology People's Hospital of Hainan Province Department of Neurology People's Hospital of Hainan Province Department of Pediatrics People's Hospital of Hainan Province

Objective:To investigate clinical and neuroimaging features of enterovirus71(EV71) related acute flaccid paralysis in patients with hand-fool-mouth ***:Nine patients with acute flaccid paralysis met the criterion of EV71 induced hand-foot-mouth disease underwent spinal and brain MR imaging from May 2008 to Sep ***:One extremity flaccid was found in four cases(3 with lower limb,1 with upper limb),two limbs flaccid in three cases(2 with lower limbs,1 with upper limbs),and four limbs flaccid in two *** MRI studies showed lesion with high signal in T2-weighted images(T2WI) and low signal T1-weighted images(T1WI) in the spinal cord of all nine cases,and the lesions were mainly in bilateral and unilateral anterior hom of cervical spinal cord and spinal cord below thoracic 9(T9) *** addition,the midbrain,pons, and medulla,which were involved in 3 cases with brainstem encephalitis,demonstrated abnormal ***,spinal cord contrast MRI studies showed mild enhancement in corresponding anterior hom of the involved side,and strong enhancement in its ventral ***: EV71 related acute flaccid paralysis in patients with hand-foot-mouth disease mainly affected the anterior hom regions and ventral root of cervical spinal cord and spinal cord below T9 level. MR imaging could efficiendy show the characteristic pattern and extent of the lesions which correlated well with the clinical features.

关键词： enterovirus infection Hand-foot-mouth disease Acute flaccid paralysis Magnetic resonance imaging

New Simian enterovirus 19(EV-A122) Strains in China Reveal Large-Scale Inter-Serotype Recombination between Simian EV-As

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Virologica Sinica 2021年第6期36卷 1652-1655页

作者： Zhenzhi Han Jinbo Xiao Yang Song Shuangli Zhu Dongyan Wang Huanhuan Lu Tianjiao Ji Dongmei Yan Wenbo Xu Yong Zhang WHO WPRO Regional Polio Reference Laboratory NHC Key Laboratory for BiosafetyNHC Key Laboratory for Medical VirologyNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing102206China Center for Biosafety Mega-Science Chinese Academy of SciencesWuhan430071China

Dear Editor,The genus enterovirus(EV),belonging to the family Picornaviridae,order Picornavirales,comprises 15 species:enterovirus A–L and rhinovirus A–C(Zell et al.2017).enterovirus A(EV-A)has 25 serotypes,including several pathogens,such as EV-A71,coxsackievirus A16(CV-A16),and CV-A6 that infect humans,as well as four serotypes(EV-A122,123,124,and 125)isolated from simians(Zell et al.2017).EV genomes share a similar structure with a length of 7.5 kb,and two open reading frames(ORFs)flanked by 5′-and 3′-untranslated regions(UTRs)in some EVs(Oberste et al.2013b;Zell et al.2017;Guo et al.2019;Lulla et al.2019).

关键词： enterovirus frames

A Functionalized Multi-walled Carbon Nanotube and Gold Nanoparticle Composite-modified Glassy Carbon Electrode for the Detection of enterovirus 71

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Biomedical and Environmental Sciences 2019年第7期32卷 541-545页

作者： CHENG Jing KOU Jing FAN Zhao Yu HAN Yi Qun MEI Yong GUO Zhen Zhong ZHAN Jian Bo School of Public Health Medical College Wuhan University of Science and Technology Wuhan 430065 Hubei China Hubei Province Key Laboratory of Occupational Hazard Identification and Control Wuhan University of Science and Technology Wuhan 430065 Hubei China Institute of health inspection and testing Hubei Provincial Center for Disease Control and Prevention Wuhan 430079 Hubei China

Hand, foot, and mouth disease (HFMD) is a common infectious disease in children, occurring primarily in preschool children[1_3] with infants under three years old being gen erally susceptible. The disease is caused by various enteroviruses, among which EV71 and Coxsackievirus A group 16 (Cox A16) are the most comm on ⑷.According to in formation released by the Chinese Center for Disease Control and Prevention on June 8, 2016, EV71 infection- related HFMD has been prevalent among infants and young children in China since 2007, with a high incidenee and many deaths.

关键词： Glassy Carbon Electrode Detection of enterovirus

Limited value of procalcitonin,C-reactive protein,white blood cell,and neutrophil in detecting bacterial coinfection and guiding antibiotic use among children with enterovirus infection

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World Journal of Pediatrics 2022年第3期18卷 230-233页

作者： Rui-Mu Zhang Kun Tan Shu Fu Ji-Kui Deng Department of Infectious Diseases Shenzhen Children’s HospitalShenzhen 518038China

Procalcitonin(PCT),C-reactive protein(CRP),and white blood cell(WBC)have been used as markers of bacterial infection in children for *** studies have suggested PCT,CRP,WBC,and percentage of neutrophils(%N)may be useful in detecting bacterial infection in children[1-4].However,elevated levels of these biomarkers have also been noted in children with enterovirus infection[5-7].In a study involving 5692 hospitalized children with herpangina or hand,foot,and mouth disease(HFMD)in two periods of years,the medians of CRP were 50.1 and 42.5 mg/L,respectively.

关键词： enterovirus infection elevated

Development of stable, cold-adapted, temperature-sensitive/conditional lethal chimeric enterovirus A71 and coxsackievirus A16

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Virologica Sinica 2022年第5期37卷 769-773页

作者： Kaw Bing Chua Qimei Ng Tao Meng Qiang Jia Temasek Life Sciences Laboratory 1 Research LinkNational University of Singapore117604Singapore

Dear Editor,Coxsackievirus A16(CA16)and enterovirus A71(EV-A71)are the main causative agents of hand foot and mouth disease(HFMD)(Alexander et al.,1994).Besides HFMD,EV-A71 infection in young children can lead to a spectrum of other clinical diseases and associated neurological complications and mortality(Melnick JL.,1996).In recent decades,EV-A71 infections have become a major public health concern throughout the world,following the frequent occurrence of epidemics and outbreaks of HFMD associated with neurological complications and high mortality in young children(Ishimaru et al.,1980;Tagaya et al.,1981;Chomnaitree et al.,1982;Samuda et al.,1987;Gilbert et al.,1988;Hayward et al.,1989;Lum et al.,1998;Chang et al.,1998;Liu et al.,2000;Yang et al.,2011;Zeng et al.,2012).

关键词： mortality enterovirus