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Proliferation Characteristics of CVA16 in KMB17 Cells under Different Culture Conditions and Analysis of Relevant Host Cell Transcriptional Features
LONG Jiaqing;LI Yingyan;YU Li;ZHENG Huiwen;LIAO Yun;WANG Jingjing;LI Heng;ZHAO Xing;ZHAO Heng;LI Dandan;ZHANG Ying;LIU Longding;Objective Hand, foot, and mouth disease(HFMD) is a common viral infectious disease in children, primarily caused by Enterovirus A71(EV-A71) and Coxsackievirus A16(CVA16). Among them, EV-A71 is closely associated with severe neurological complications and death. With the widespread use of the EV-A71 vaccine, CVA16 has gradually become a major causative pathogen, highlighting the urgent need to establish an efficient large-scale vaccine production process. Methods In this study, CVA16 was serially passaged 30 times in KMB17 diploid cells under different temperature conditions(33℃, 37℃, and 38.5℃) and multiplicities of infection(MOIs: 0.01, 0.1, and 0.5). Viral growth kinetics, virion integrity, and genetic stability were systematically evaluated. In addition, single-cell transcriptomic analysis was performed to characterize host cell response mechanisms, with the aim of providing a scientific basis for optimizing culture parameters, improving viral yield and quality, and supporting vaccine production. Results The results showed that 37℃ and an MOI of 0.1 were the optimal amplification conditions. Under these conditions, viral replication peaked at 24 – 36 h, with titers exceeding 108.0 CCID50, and intact virions accounting for 78% of total viral particles. Genomic sequencing confirmed that the mutation rate in the VP1 region remained below 0.1%involving growth factor-related genes such as CXCL8 and IER3, as well as the endoplasmic reticulum protein-processing pathway, were conducive to viral replication. At later stages, further upregulation of genes such as IL6 and LDHA suggested that CVA16 achieved efficient replication by modulating host translation and the cell cycle. Conclusion This study demonstrates that culture at 37℃ with an MOI of 0.1 is favorable for CVA16 replication in KMB17 diploid cells and provides key process parameters for the large-scale production of CVA16.
Pathogenic Spectrum and Genetic Characteristics of Hand, Foot and Mouth Disease in Kunming, Yunnan Province, China, 2022–2023
LIU Xuanguoer;JIAGN Lili;CUN Jianping;YANG Xi;FU Xiaoqing;QI Yanbo;TIAN Bingjun;Objective To determine the pathogenic spectrum of hand, foot and mouth disease(HFMD) in Kunming, Yunnan Province, China,from 2022 to 2023, and to analyze the VP1 gene characteristics of common pathogens, including Coxsackievirus A4(CVA4), CVA6, CVA10, and CVA16. Methods A total of 161 stool samples from HFMD cases collected in Kunming during 2022-2023, which were positive for EV-A71, CVA16 or pan-enterovirus detected by real-time fluorescent quantitative PCR, were selected for molecular typing.. Viral genotypes were first determined by sequencing the VP4/VP2 junction region. Complete VP1 gene sequences were subsequently obtained for each viral type. Representative VP1 sequences of CVA4, CVA6, CVA10, and CVA16 were downloaded from published databases, and phylogenetic trees were constructed using MEGA version 6.0 to analyze genetic characteristics. Results All 161 samples were successfully amplified and genotyped, yielding a sequencing success rate of 100%(161/161). The most frequently detected viruses were CVA6(48 strains, 29.81%), CVA4(39 strains, 24.22%), CVA10(37 strains, 22.98%), and CVA16(23 strains, 14.30%), all of which belong to enterovirus species A. Fourteen additional strains were identified at lower frequencies, including CVA2(1 strain), CVB5(7 strains), echovirus 3(2 strains), poliovirus type 1(2 vaccine strains), and poliovirus type 3(2 vaccine strains). Genetic analysis of the complete VP1 region showed that all CVA6 strains belonged to subgenotype D3a, CVA4 strains to subgenotype C2, CVA10 strains to genotype C, and CVA16 strains to subgenotype B1a. Conclusion From 2022 to 2023, the predominant pathogens causing HFMD in Kunming were CVA6, CVA4, CVA10, and CVA16. The genotypes and subgenotypes of these major circulating viruses were consistent with those reported in other regions of China, suggesting long-term persistence and circulation of these viral lineages nationwide. These findings highlight the importance of continued etiological surveillance and molecular epidemiological studies of HFMD in Kunming.
Isolation of Human Metapneumovirus from a Hospitalized Child and Characteristics of its Infection in Human Respiratory Epithelial and Alveolar Organoids
SHI Xiaoya;LU Roujian;ZHAO Yang;YU Yufan;ZHAI Chengcheng;NIU Peihua;SONG Jingdong;DENG Yao;TAN Wenjie;ZHU Na;Objective To isolate human metapneumovirus(hMPV) from a pediatric patient with an unusually prolonged respiratory infection, analyze its in vitro biological characteristics, and evaluate its susceptibility to broad-spectrum antiviral agents, thereby assessing the risk of its further transmission leading to severe infection in infants. Methods Human metapneumovirus was isolated from the patient's throat swab using an air-liquid interface human airway epithelial organoid(HAE) model. Viral replication kinetics were determined. Viral morphology was examined by electron microscopy, and expression of the hMPV N protein in HAE was detected by immunofluorescence. The full-length genome of the isolated virus was sequenced and phylogenetically analyzed. The isolated virus was used to infect alveolar organoids and conventional LLC-MK2 cells to measure viral load and observe virus-induced cytopathic effects. Meanwhile, the susceptibility of this hMPV isolate to the broad-spectrum antiviral drug ribavirin was validated in both airway organoids and LLCMK2 cells. Results A clinical isolate of human metapneumovirus, designated hMPV-CTan01/Beijing/2023(hereinafter referred to as hMPV-CTan01), was obtained from the throat swab of the pediatric patient with a prolonged disease course. Its complete genome is 13,435 bp in length, belonging to the hMPV A2C subgenotype, showing high consistency with currently circulating strains. hMPV-CTan01 efficiently replicated in HAE and produced infectious progeny virus. Typical hMPV morphology was observed in the HAE supernatant, and N protein expression was detected in airway epithelial cells. hMPV-CTan01 was able to infect alveolar organoids, suggesting its potential to cause severe infection. Furthermore, hMPV-CTan01 replicated efficiently in LLC-MK2 cells, inducing typical cytopathic effects, and its in vitro characteristics were consistent with previous reports. The antiviral drug ribavirin suppressed the replication of this isolate in both LLC-MK2 cells and HAE. Conclusion The in vitro biological characteristics of hMPV-CTan01, isolated from the respiratory tract of this pediatric patient with a prolonged illness, are consistent with previous reports and do not demonstrate enhanced pathogenicity in vitro. Ribavirin effectively inhibits its replication in both airway organoids and LLC-MK2 cells, indicating no apparent drug resistance. These findings demonstrate that although hMPV-CTan01 was associated with a prolonged disease course in this patient, its risk of further transmission leading to severe infection is low.
Whole Genome Sequencing and In Vitro Host Transcriptome Remodeling Analysis of Clinical Isolates of Human Adenovirus B3 and B7 Associated with Severe Pneumonia in Children
LI Ruorui;LU Roujian;GUO Xiaojuan;ZHAO Zhihao;SHEN Jun;WU Changcheng;ZHAI Desheng;TAN Wenjie;Objective To investigate the differences between human adenovirus HAdV-B3 and HAdV-B7 in replication kinetics, genetic characteristics, and host-cell transcriptomic remodeling strategies, and to identify candidate host-associated factors for subsequent antiviral studies. Methods The growth trends of the two viral types were compared in MRC-5 cells. Whole-genome sequencing and evolutionary analysis were performed on six clinical isolates. Transcriptome sequencing(RNA-seq) was used to analyze the global transcriptional landscape of MRC-5 cells at 16 h post-infection(hpi). Changes in the expression levels of candidate host factors were further validated by RT-qPCR. Results In MRC-5 cells, HAdV-B3 exhibited a shorter replication initiation period and an earlier trend toward peak viral titer than HAdV-B7. Whole-genome analysis showed that all six isolates belonged to the HAdV-B lineage. The proportion of viral transcripts was significantly higher in HAdV-B3 than in HAdV-B7(10.9% vs. 1.1%). Transcriptomic analysis showed that HAdV-B3 caused a markedly greater disturbance of host transcriptional homeostasis than HAdV-B7. Functional enrichment analysis revealed that both viruses hijacked host DNA replication and cell-cycle regulatory programs and induced an antiviral response centered on IFN-β and ISG15. Among them, HAdV-B3 showed a stronger regulatory effect on pathways associated with coordinated upregulation of the G1/S transition(E2F8, CCNE1, and PCNA) and replication stress response(TP73). Conclusion Both HAdV-B3 and HAdV-B7 induced significant host transcriptomic reprogramming in MRC-5 cells, whereas HAdV-B3 showed stronger replication activity, cytopathic ability, and host transcriptional regulatory effects. Genes including TP73, IFN-β, ISG15, CXCL10, E2F8, CCNE1, PCNA, TK1, and RRM1 are associated with host antiviral responses and cell cycle/DNA replication remodeling, and may serve as candidate host factors for subsequent mechanistic studies and screening of drug targets.
Whole Genome Analysis of Human Parainfluenza Virus Type 4 in Beijing,China,2025
GAO Bin;ZHANG Weijia;WU Dan;XU Hui;LIU Yimeng;SHI Weixian;LU Guilan;CUI Shujuan;ZHANG Daitao;ZHAO Jiachen;Objective To characterize the genetic features of human parainfluenza virus type 4(HPIV4) circulating in Beijing in 2025. Methods A total of 21,002 throat swab samples from influenza-like illness(ILI) cases and 7,570 throat swab or lower respiratory tract specimens from severe acute respiratory infection(SARI) cases were collected in Beijing in 2025 and screened for HPIV4. Whole-genome sequencing was performed on HPIV4-positive samples with cycle threshold(Ct) values <30. The complete viral genomes were amplified using multiplex PCR. Nucleotide and amino acid variations in the hemagglutinin – neuraminidase(HN) and fusion(F) genes were analyzed using MEGA 6.0. Phylogenetic trees based on whole genomes, HN, and F genes were constructed using MEGA 6.0. Potential N-glycosylation sites in HN and F proteins were predicted using the NetNGlyc 1.0 server, and recombination analysis was conducted using SimPlot 3.5.1. Results The overall positivity rate of HPIV4 was 0.31%(81/25,872), with rates of 0.30%(63/21,002) in ILI cases and 0.24%(18/7,570) in SARI cases. Viral activity peaked in summer and autumn. Nineteen complete HPIV4 genomes were obtained. The predominant subtype was HPIV4b, mainly belonging to clade C(78.95%, 15/19), while HPIV4a sequences were primarily assigned to clade C3(21.05%, 4/19). The genomes(average sequencing depth >1,100×) were approximately 17.3 kb in length, with Q30 values >85% and GC content ranging from 35.9% to 36.3%. For HPIV4b, nucleotide identities of the HN and F genes ranged from 98.28%– 100.00% and 97.81% – 100.00%, respectively, whereas for HPIV4a they ranged from 89.26% – 99.93% and 99.51% – 99.82%. Variation analysis identified subtype-specific amino acid substitutions in the F protein. Both HN and F genes were predominantly under purifying selection. Five potential N-glycosylation sites were identified in the HN protein and three in the F protein. No recombination events were detected in either gene. Conclusion In 2025, HPIV4 circulation in Beijing showed subtype-specific temporal patterns, with HPIV4b as the dominant subtype. The HPIV4 genome exhibits subtype-specific amino acid variation, with evolution primarily driven by purifying selection and no evidence of recombination. Glycosylation sites display both conserved and subtype-specific features. These findings provide important genomic data to support long-term surveillance and control strategies for HPIV4 in Beijing.
Research Progress on the Pathogenic Mechanisms and Prevention and Control Strategies of Nipah Virus
MENG Yu;ZOU Min;CHEN Bin;ZHANG Fengxue;WANG Wenlu;YANG Zifeng;Nipah virus(NiV) is a highly pathogenic zoonotic virus maintained in nature by fruit bats of the genus Pteropus. NiV infection can cause outbreaks characterized by acute encephalitis and/or severe respiratory disease, accompanied by household clusters, nosocomial transmission, and close-contact-associated human-tohuman spread, posing a substantial public health threat. Case fatality rates reported in different outbreaks are often 40% – 75%, and their variability is associated with factors including healthcare accessibility, case composition and selection bias, outbreak scale, as well as laboratory confirmation and surveillance capability. Currently, no NiV-specific therapeutics or vaccines have been approved, and clinical management mainly relies on supportive care and infection prevention and control; therefore, countermeasure development has been prioritized internationally. In recent years, studies on NiV molecular pathogenesis have focused on receptormediated entry and membrane fusion, the replication – transcription complex, and interferon antagonism mediated by P gene-encoded proteins, providing important clues for antiviral interventions. This review summarizes recent advances in NiV infection and pathogenesis, evaluates the current status and major translational challenges of antivirals, passive immunization, and vaccine strategies, and discusses future directions in target validation, product development, and the establishment of “One Health” surveillance and control systems.
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Epidemiology of Foot-and-Mouth Disease and Advances in Diagnostic Technologies
ZHOU Zheng;SHI Zhengwang;LIAO Huancheng;LUO Juncong;WU Jiahui;YANG Qianqian;WANG Lin;XIE Yage;TIAN Hong;ZHENG Haixue;Foot-and-mouth disease(FMD) is a highly contagious and transboundary viral disease affecting cloven-hoofed animals, caused by foot-and-mouth disease virus(FMDV). FMDV comprises seven serotypes and more than 65 topotypes, with marked differences in epidemiological characteristics among serotypes. Globally, serotypes O and A are the most widely distributed, and the overall epidemiological pattern of FMD is characterized by sporadic outbreaks and high genetic diversity. In recent years, the transmission dynamics of FMDV have undergone significant changes, with traditional control barriers based on geographic isolation increasingly being breached. For example, Hungary reported a resurgence of FMD in 2025 after a 52-year absence, without epidemiological links to neighboring countries. In addition, South African Territories(SAT) strains have expanded beyond their traditional endemic regions, showing continued spread toward West Asia and the Middle East, thereby further complicating global control efforts. Recent outbreaks are frequently associated with challenges in source tracing, underscoring the urgent need to refine and update current control strategies. Accurate and sensitive diagnostic technologies play a central role in early warning, outbreak investigation, and source tracing, and are critical for risk assessment and targeted control. This review systematically summarizes the global epidemiological patterns and characteristics of FMDV and provides an overview of current diagnostic methods. It also discusses key technical challenges, including antibody level evaluation and serological differentiation between vaccinated animals and those infected with wild-type virus, with the aim of providing a scientific basis and practical guidance for FMD prevention and control.
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Research Progress on Gosling Gout Associated with Goose Astrovirus Genotype 2
SUN Kaiyun;QIU Yang;ZHANG Xuezheng;ZHAO Rui;FU Kang;PANG Wenshuang;QIU Jianhua;WANG Gang;LI Ning;Gosling gout caused by goose astrovirus genotype 2(GAstV-2) is an infectious disease characterized by urate deposition in the visceral organs and joints of goslings. The disease mainly affects goslings younger than 3 weeks of age and is associated with high morbidity and mortality. Surviving goslings often exhibit growth retardation, immunosuppression, and increased susceptibility to secondary bacterial infections. In recent years, the disease has become widely prevalent in major goose-producing regions of China, causing substantial economic losses to the goose industry. GAstV-2 has also been confirmed to infect Cherry Valley ducks, Muscovy ducks, and chickens, indicating a potential risk of cross-species transmission and posing a broader threat to the poultry industry. Owing to the incomplete understanding of its pathogenic mechanisms, together with the frequent genetic variation among GAstV-2 strains and the common occurrence of mixed infections in clinical settings, no commercial vaccine is currently available. Therefore, this review systematically summarizes recent advances in the etiology, epidemiology, pathogenic mechanisms, diagnostic techniques, and prevention and control strategies of GAstV-2-associated gosling gout, with the aim of providing a theoretical basis for disease surveillance and early warning, further elucidation of pathogenic mechanisms, and the scientific development of prevention and control strategies.
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Screening and Identification of Key Interacting Proteins Regulated by Ring Finger Protein 5 During African Swine Fever Virus Infection
LIU Ying;SUN Hualin;CHEN Yikang;LAN Xinting;YANG Jifei;GUAN Guiquan;DU Shuzeng;NIU Qingli;YIN Hong;Objective African swine fever(ASF) is a highly lethal infectious disease caused by African swine fever virus(ASFV), resulting in severe hemorrhagic symptoms and high mortality in domestic pigs and wild boars. Ring finger protein 5(RNF5) is an E3 ubiquitin ligase involved in ubiquitin-mediated protein degradation and inflammatory signaling pathways. This study aimed to investigate the role of RNF5 and its mediated host protein interaction network during ASFV infection. Methods The effect of RNF5 overexpression on ASFV replication was first evaluated in vitro. Subsequently, co-immunoprecipitation coupled with mass spectrometry(Co-IP-MS) was performed to identify RNF5-interacting proteins, followed by functional analysis of differentially enriched proteins. Finally, candidate interacting proteins were further validated by coimmunoprecipitation assays under ASFV infection conditions. Results RNF5 overexpression significantly inhibited ASFV replication, indicating its potential antiviral activity. Co-IP-MS analysis identified multiple candidate proteins potentially interacting with RNF5. Functional analysis indicated that these proteins were mainly involved in biological processes including transcription and RNA processing, metabolism, and signal transduction. Co-immunoprecipitation assays further confirmed that RNF5 specifically interacted with DEADbox helicase 3 X-linked(DDX3X) and B-cell lymphoma-2(BCL-2). Under ASFV infection conditions, the expression level of DDX3X was significantly upregulated, whereas BCL-2 expression showed no obvious change; however, the interactions of both proteins with RNF5 were significantly enhanced following ASFV infection. Conclusion This study confirms the inhibitory effect of RNF5 on ASFV replication and further characterizes its potential host interaction network. The findings provide new insights into how ASFV may modulate host protein interaction networks to influence viral replication and host cellular responses.
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Residual Risk of Hepatocellular Carcinoma after Clinical Cure of Chronic Hepatitis B
GAO Zixuan;HAO Hongxiao;YAO Linmei;WEI Xin;WANG Shuojie;CAO Weihua;ZHANG Yaqin;DENG Wen;WANG Shiyu;LI Xinxin;ZHANG Ziyu;QIU Dianya;MA Tianyu;LI Minghui;Chronic hepatitis B(CHB) remains a major global cause of hepatocellular carcinoma(HCC). With advances in nucleos(t)ide analogue( NA) and interferon( IFN) therapies, an increasing proportion of patients can achieve sustained hepatitis B virus DNA(HBV DNA) suppression and even hepatitis B surface antigen(HBs Ag) clearance. In this review, clinical cure(also referred to as functional cure) is defined by HBs Ag seroclearance, ideally sustained for at least 12 months after treatment cessation, and is distinguished from virological suppression characterized by undetectable HBV DNA with persistent HBs Ag positivity. Importantly, clinical cure does not eliminate the risk of HCC. Residual risk remains and is influenced by factors including prior cirrhosis, advanced age, male sex, HBV genotype C, and a family history of HCC. NAs reduce HCC incidence by approximately 50% through durable suppression of viral replication but have limited capacity to induce HBs Ag clearance. In contrast, IFN exerts additional benefits by inhibiting the oncogenic activity of hepatitis B virus X protein( HBx), modulating the immune microenvironment, and promoting HBs Ag decline. Recent advances in combination therapies, including IFN with NAs or RNA interference – based agents—particularly small interfering RNA( si RNA) and antisense oligonucleotides( ASOs)—have markedly improved clinical cure rates and are considered among the most promising strategies for reducing long-term HCC risk. Future efforts should focus on establishing post-cure risk stratification and surveillance strategies to enable a transition in disease management from“ viral suppression” to“ sustained cancer prevention”.
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Isolation of Subgroup J Avian Leukosis Viruses and Their Partial Sequence Comparison
DU Yan 1, CUI Zhi zhong 1,2 , QIN Ai jian 1, Silva R. F. 3, Lee L. F. 3 (1.Department of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; 2. College of Animal Science, Shandong Agricultural University, Taian 271018, China; 3.Subgroup J avian leukosis viruses (ALV J) were isolated from two broiler breeder farms with suspected diseased chickens and two commercial broiler flocks without clinical symptoms by inoculating the samples into chicken embryo fibroblast cells and PCR amplification of the infected CEF genomic DNA. In the indirect fluorescence assay (IFA) with ALV J specific monoclonal antibody JE9, 2 strains, SD9901 and SD9902 from breeder with suspected lesions, were strongly positive, another 2 strains, YZ9901 and YZ9902 from commercial broilers without clinical symptoms gave weak reactions. The genomes of strains YZ9901 and SD9902 were partially sequenced and the results indicated that their gp85 had 89%-93% identity in the amino acid level with ALV J prototype HPRS 103 and American strain ADOL HCl. The amino acid identity among themselves was 92%. The 3' noncoding LTR region had 95%-97% identity in the nucleotide level with ALV J prototype strain HPRS 103. But the Chinese strains had a 139 base deletion mutation in their E elements nearby the 3' LTR region and got an insertion of 11 base fragment instead.
CONSTRUCTION AND APPLICATION OF A HIGH LEVEL EXPRESSION VECTOR CONTAINING P_RPL PROMOTER
Zhang Zhiqing Yao Lihong Hou Yunde(National Laboratory of Molecular Virology and Genetic Engineering, Institute of Virology, Chinese Academy of Preventive Medicine, Beijing )A high level expression vector has been constructed,which contains PR PL promoter, cIts857 gene, multiple cloning sites ( MCS ) and two strong transcription terminators. Foreign gene with ATG signal can be inserted into the MCS, expressing non-fusion protein. Using this vector, we have expressed successfully the human interferon r,human interleukin-2 and human tumor necrosis factor. The foreign gene product accounts for more than 20% of the total cell protein.
NORWALK-LIKE VIRUS INFECTION FOUND IN DIARRHEA PATIENTS IN CHINA
Fang Zhaoyin Wen Leying Jin ShengJin Zhao Zhanghua C. Moe H. Yoshikura R.Glass(Institute of Virology,CAPM ,Beijing 100052) 1.Henan Institute of Traditional Chinese Medicine; 2.Centers for Disease control and prevention,USA; 3.University of Tokyo,JapanNorwalk virus or Norwalk virus -like agents are important pathogens that causeoutberaks of acute nonbacterial gastroenteritis. Two Norwalk-like virus isolates were identifiedin fecal specimens from acute diarrhea patients in Henan province during Oct.1990-Jan1991 rotavirus season by electron microscopy that shows 28nm diameter with structured capsid.RT-PCR using Norwalk -specific primer pair 51-3,and nucleotidesequencing of PCR products showed 72%homology of these two isolates to that of Norwalkvirus prototype 8FⅡa. Our finding suggests that more attention should be pai to outbreaks of acute gastroenteritis caused by Norwalk-like virus in China.
Identification of a New Subgroup of Avian Leukosis Virus Isolated from Chinese Indigenous Chicken Breeds
WANG Xin,ZHAO Peng,CUI Zhi-zhong(College of Veterinary Medicine,Shandong Agricultural University,Tai an 271018,China)In order to clarify Avian leukosis virus(ALV) characteristics from Chinese native chicken breeds,three ALV JS11C1,JS11C2 and JS11C3 were isolated from Chinese native breed "luhua" by inoculation of DF1 cell culture and detection of p27 antigen.Using PCR amplification of env gene,the amplified gp85 genes were analyzed and compared to all six chicken ALV subgroups reported.The gp85 genes of these three viruses were 1 005bp in length and encoded 335 amino acids,and the gp37 genes were 609bp and encoded 203 amino acids.The homology of gp85 among these three isolated strains was 91.9%-97.0%.Comparing to 18 stains of subgroup A,B,C,D,E published in GenBank,the homology was only in the range of 77.7%-84.6%,significantly lower than the gp85 homology observed within the common chicken subgroups A(88.2%-98.5%),B(91.6%-98.8%),and E(97.9%-99.4%).The gp85 homology compared with subgroup J was only 34.2%-36.5%.These results suggested that three isolated strains from Chinese native breed "luhua" belong to a new subgroup different from all six known subgroups from Chickens,and thus designated as subgroup K.
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Identification of Enterovirus Type 71 by RT-PCR and the Gene Characterization
CUI Ai-li~ 1, XU Wen-bo~ 1, LI Xiu-zhu~ 2, HU Jia-yu~ 2, LING Hua~ 3, TANG Wei~ 2, YANG Zhi-hong~ 4, ZHANG Yan~ 1, CHEN Li~ 1, Hiroyuki Shimizu~ 5(1. National Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052, China;2. Shanghai Center for Disease Control and Prevention, Shanghai 200336, China;3. Chongqing Center for Disease Control and Prevention, Chongqing 400042, China;4. Childrens Hospital, Fudan University, Shanghai 200032, China;5. National Institute of Hygiene, Japan)10 virus strains were isolated from clinical specimens of children with hand-foot-and-mouth disease (HFMD), 9 isolates from Shanghai and 1 isolate from Chongqing. All of the 10 isolates were tested by RT-PCR assay with two specific primer pairs for VP1 genes of enterovirus type 71 (EV71) and Coxsackie virus A16 (Cox. A16) respectively. The enterovirus serotype 71 and Cox. A16 were primarily identified depending on the size of PCR products and the primers used. The RT-PCR results indicated that 2 EV71 isolates were from Shanghai, 1 was from Chongqing and 7 Cox. A16 isolates were from Shanghai. All of the 10 PCR products were sequenced, the sequence analysis confirmed that PCR identified results was 100% correlative to virus sequencing, so RT-PCR assay is highly specific and probably may be the first choice for identification of EV71 and Cox. A16. 891 nucleotides of VP1 coding genes of 3 EV71 isolated strains were sequenced and compared with that of previously isolated 7 EV71 Chinese isolates available from GenBank (SHZH03?SHZH98?SH-F1?SH-F2?SH-H25?SH-H26 and CHN-87) by homogeneity and phylogenetic tree analyses. The homogeneity of these 10 Chinese strains with the representative isolates of C genotype of EV71 was between 89.3%-94.6%; with the representative isolates of A and B genotypes was between 81.3%-84.0%. The data suggested that all of the 10 Chinese isolates belong to EV71 genotype C except CHN-87, which was untyped. The homogeneity of the 3 EV71 isolated strains and 6 previously isolated strains (SHZH03?SHZH98?SH-F1?SH-F2?SH-H25?SH-H26) were between 94.5%-100%, that formed a single branch in the phylogenetic tree. There were only 89.3%-92.9% homology among these 9 strains and the representative strains of C1?C2?C3 sub-genotypes of EV71, this suggested that these 9 Chinese isolates and the TAI-98 composed a new sub-genotype, the C4 sub-genotype, of the C genotype. EV71 of sub-genotype C4 distributed in Shenzhen, Chongqiang and Shanghai from 1988-2003. It is much helpful to develop EV71 diagnosis, virus surveillance, virus standard nomenclature and set up EV71 virus bank and virus gene bank to accelerate the control and prevention of EV71 outbreak in China and in the world.
Research Progress on the Infection Mechanism of Coronavirus SARS-CoV-2
LU Rongguang;WU Jing;BAI Xue;LIU Weiquan;An emerging infectious disease COVID-19 which caused by a novel coronavirus SARS-CoV-2,has spread to many countries and regions around the world. For now,COVID-19 triggered a global public concern about healthy safety. Although just about 2% SARS-CoV-2 infected patients have contacted with wild animals,most scientists still believe that SARS-CoV-2 origin from wild animals. In fact,virus interspecies transmission is very difficult and lead to the new host dead in most cases. However,different from the other viruses,SARSCoV-2 have adapt to human body very well since SARS-CoV-2 emerged. Thus,we reviewed several research advances about etiology,function receptor and evolution of SARS-CoV-2,try to provide a new perspective to understand the emergence of SARS-CoV-2.
Prediction of the Epidemic Trend of COVID-19
YAN Mingjiang;DONG Yihong;JIA Xiangen;ZHENG Haiyang;XIN Yu;Coronavirus disease 2019(COVID-19) spread initially from Wuhan(Hubei Province,China) in December 2019 through China,but is now a pandemic. Unprecedented steps have been taken throughout China to vigorously carry out disease treatment and epidemic prevention. Official statistics published by the National Health Commission of the People's Republic of China were collected to predict the trend of the epidemic. In the traditional Susceptible,Exposed,Infectious,Recovered(SEIR) model,only infectious patients and noninfectious latent patients are considered. However,COVID-19-diagnosed patients cannot infect the susceptible population because they have been isolated in hospitals,whereas latent patients may be infectious. Based on this information,we propose an improved model of infectious-disease transmission:"ISEIR". In ISEIR,patients are divided into outpatients(with infectivity)and inpatients(infectivity is not considered). Preclinical patients who are infectious are also considered. ISEIR fits model parameters dynamically with historical data to exclude the limitations of fixed parameters. The data of patients diagnosed early with COVID-19 in Hubei Province,China was seriously distorted. Therefore,according to the probability distribution of the daily basic reproduction number(R0),the clinical-diagnosis data of February 12–14 were preprocessed and spread into previous data to correct distortion of previous data. The epidemic situation was divided into two regions:the whole country(excluding Hubei Province,China)and Hubei Province,China. The new ISEIR predicts further development of the future epidemic,and calculates the change in daily R0. Results revealed that the R0 of Hubei Province,China has reduced gradually from 3.108. All patients will be cured and discharged from hospital around April 19.The initial R0 of China(excluding Hubei Province)was 1.929,and all patients will be cured around March 26.Results showed that the epidemic has been suppressed effectively under strict prevention-and-control measures.It is also necessary to prevent rebound of the epidemic situation caused by the resumption of employment.
Research Progress in Novel Coronavirus(2019-nCoV)-Related Drugs In Vitro and In Vivo
SONG Gao;CHENG Mengqun;WEI Xianwen;In December 2019 in Wuhan City(Hubei Province,China),multiple cases of patients with pneumonia infected by a new type of coronavirus were noted. With the spread of the epidemic,other cases in China and overseas have also been found. On 12 January 2020,the World Health Organization tentatively named it"2019 Novel Coronavirus"(2019-nCoV). This is a new type of virus,which is highly infectious and can cause severe respiratory diseases. A clinically efficacious treatment is lacking. We reviewed the guidelines for recommended therapeutic drugs and drug-development advances with the aim of providing a reference for clinical treatment of 2019-nCoV infection.
Research Progress on SARS-CoV-2
XIE Qian;WU Zhengyu;SHU Yuelong;Since December 2019,the outbreak of coronavirus disease 2019(COVID-19)in Wuhan,China,has spread rapidly to other provinces and cities in China,and worldwide. Severe acute respiratory syndrome(SARS)-CoV-2 belongs to the β-coronavirus family,which is closely related to SARS-CoV and Middle East respiratory syndrome(MERS)-CoV,but quite different,especially in the spike protein. SARS-CoV-2 may be derived from bats according to sequence comparison. SARS-CoV-2 uses the same receptor,angiotensin converting enzyme Ⅱ(ACE2),as SARS-CoV. The main transmission routes include droplets and close contacts. The lack of effective drugs and vaccine is a challenge for outbreak control.
Research Progress of the Molecule Mechanisms of Ebola Virus Infection of Cells
SHI Ming,SHEN Yu-qing(Medical School of Southeast University,Nanjing 210009,China)Ebola virus can cause severe Ebola hemorrhagic fever.The mortality rate is 90 percent.Up till now,there is no effective vaccine or treatment of Ebola virus infection.Relaed researches on Ebola virus have become a hot topic in virology.The understanding of molecular mechanisms of Ebola virus infection of cells are important for the development of vaccine and anti-virus drugs.Therefore,this review summarized the recent research progress on the mechanisms of Ebola virus infection.
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