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Identification and Mechanistic Characterization of TMEM240 as a Key Host Factor for Echovirus 30
LI Jichen;DUAN Wei;LI Huijie;WANG Rui;SUN Qiang;ZHANG Yong;Objective Echovirus 30(E30), a prominent member of the Enterovirus B species, is one of the major causative agents of aseptic meningitis and encephalitis and exhibits seasonal and cyclical outbreaks in Asia, Europe and South America. However, the host factors and molecular mechanisms governing E30 infection remain poorly understood. The CRISPR/Cas9 genome-editing system provides a powerful approach for dissecting host gene function during viral infection. This study aimed to identify and characterize host factors essential for E30 replication. Methods Based on host factors identified in a prior genome-wide screen, CRISPR/Cas9-mediated gene editing was used to generate a knockout targeting the intracellular loop-encoding exon of transmembrane protein 240(TMEM240). Complete ablation of TMEM240 protein expression was confirmed by Western blotting in clonal human rhabdomyosarcoma(RD) cells. The impact of TMEM240 deficiency on E30 infection and replication was subsequently investigated. Results Knockout of TMEM240 significantly inhibited E30 replication at the early stage of infection, while viral binding and internalization were not affected. Immunofluorescence analysis demonstrated markedly reduced co-localization of E30 doublestranded RNA with the early endosome marker Rab5A in TMEM240-deficient cells, indicating a role for TMEM240 in early endosomal replication. Transcriptomic analysis revealed that TMEM240-associated differentially expressed genes were enriched in pathways related to autophagosome maturation and autophagosome–lysosome fusion. Furthermore, mCherry–GFP–LC3B adenoviral reporter assays, Western blotting, confocal microscopy, lysosomal pH measurements, and transcription factor EB(TFEB) nuclear translocation assays showed that loss or reduction of TMEM240 impaired autophagosome – lysosome fusion, leading to autophagosome accumulation and enhanced nuclear translocation of TFEB. Conclusion These findings identify TMEM240 as a proviral host factor that facilitates early E30 replication by regulating autophagosome–lysosome fusion. This study provides new mechanistic insight into E30–host interactions and advances our understanding of the molecular basis of E30 infection.
Analysis of HFMD Pathogen Spectrum and Genetic Evolution in Guang'an City, Sichuan Province,China, 2024
LIU Xuan;YIN Liu;CEN Yi;QU Yingli;SHI Qiangqiang;CHEN Ze;MEI Guoyong;DU Haijun;YU Shuhai;SHI Hangjiu;XIA Zhiqiang;SONG Qinqin;HAN Jun;Objective To investigate the pathogen spectrum of hand, foot, and mouth disease(HFMD) in Guang'an, Sichuan Province, China, in 2024 and to characterize the genetic evolution of circulating enteroviruses, providing evidence for local HFMD prevention and control. Methods Pharyngeal swab samples were collected from clinically diagnosed HFMD cases in Guang'an in 2024. Samples were screened for enterovirus(EV) using quantitative real-time polymerase chain reaction(qPCR). VP1 genes from EV-positive samples were amplified by polymerase chain reaction(PCR) and subsequently subjected to sequencing, genotyping, and phylogenetic analysis. Results A total of 94 pharyngeal swab samples were collected from HFMD cases, with an EV positivity rate of 76.6%(72/94). Using universal enterovirus typing primers, 60 samples were successfully genotyped, including 5 Coxsackievirus A4(CVA4, 8.3%), 4 Coxsackievirus A5(CVA5, 6.7%), 24 Coxsackievirus A10(CVA10, 40.0%), 26 Coxsackievirus A16(CVA16, 43.3%), and 1 Echovirus 25(E25, 1.7%). Complete VP1 gene sequences were obtained from 22 strains, including 1 CVA4 strain(genotype D), 1 CVA5 strain(genotype D), 7 CVA16 strains(4 B1a and 3 B1c subgenotypes), and 13 CVA10 strains(genotype C2). Conclusion Among HFMD cases treated at Guang'an People's Hospital in 2024, CVA16 was the predominant pathogen, followed by CVA10. The main circulating lineages were B1a and B1c for CVA16 and genotype C2 for CVA10. As this study was conducted at a single center, the findings reflect only the etiological characteristics of patients attending this hospital and may not fully represent the epidemiological pattern of HFMD across Guang'an. Continuous multi-center surveillance is needed to better understand the distribution of circulating strains and support evidence-based HFMD prevention and control strategies.
Isolation, Identification, and Biological Characterization of Coxsackievirus A4 Strains
REN Xiaohong;LIAO Yun;WANG Jingjing;LI Heng;ZHAO Xin;ZHAO Heng;LI Dandan;YU Li;QIU Heng;ZHENG Huiwen;LIU Longding;Objective Hand, foot, and mouth disease(HFMD) is caused by a diverse group of enteroviruses. The isolation, identification, and characterization of Coxsackievirus A(CVA) strains are increasingly important for understanding HFMD pathogenesis and for vaccine development. This study aimed to isolate and identify Coxsackievirus A4(CVA4) strains and to analyze their biological characteristics to provide a basis for future vaccine development. Methods CVA4 was isolated using KMB17 cells. The VP1 gene of the isolates was amplified by PCR and sequenced for genotyping. Viral morphology was examined by transmission electron microscopy. Major structural proteins were analyzed by SDS-PAGE, Western blotting(WB), and silver staining. Viral replication kinetics in KMB17 cells and infection characteristics in suckling mice were investigated. Immunogenicity was further evaluated using neutralization assays and ELISpot assays. Results Three CVA4 strains were isolated from fecal samples of HFMD patients. Typical cytopathic effects(CPE), including cell shrinkage, rounding, and swelling, were observed after infection of KMB17 cells. VP1 gene sequencing confirmed that all isolates belonged to the D2 subgenotype of CVA4. Transmission electron microscopy showed typical icosahedral viral particles with diameters of approximately 30 nm, predominantly appearing as solid particles. Western blot and silver staining analysis demonstrated the presence of structural proteins VP1 and VP3 in purified viral preparations, which were recognized by specific anti-CVA4 sera. Plaque purification yielded three clonal strains designated E-4, E-2, and B-2. Growth kinetics analysis showed that all three strains replicated efficiently in KMB17 cells at a multiplicity of infection(MOI) of 0.05 and reached peak titers within 96 h, with infectious titers of 107.5, 105.75, and 107.75 CCID50/mL, respectively. Intracranial inoculation of 2-day-old suckling mice with 2000 CCID50 per mouse resulted in paralysis and death within 3–5 days post-infection for all three strains. Immunogenicity analysis indicated that intraperitoneal immunization of BALB/c mice with the three strains induced strong humoral and cellular immune responses. After two immunizations, the geometric mean neutralizing antibody titers reached 2622.2, 1456.1, and 5045.1, respectively.Conclusion Based on viral replication characteristics and immunogenicity, strains E-4 and B-2 exhibited relatively superior biological properties compared with E-2. These strains may represent promising candidates for further evaluation as potential vaccine strains.
Assessment of Immune Efficacy and Durability of an Inactivated EV ‑ A71 Vaccine in Children
WANG Jianxing;ZHANG Xiaoli;LIU Xiaolin;DONG Zhen;SUN Wenkui;KOU Zengqiang;This study investigated the seropositivity rate, geometric mean titers(GMTs), and persistence of enterovirus A71(EV-A71) neutralizing antibodies among children in Linyi City, Shandong Province,China, following administration of the EV-A71 vaccine, with the aim of evaluating its real-world protective efficacy. Using stratified random sampling, 195 children under seven years of age who had received two doses of the vaccine were recruited as the vaccination group, while 195 unvaccinated children without a history of hand, foot, and mouth disease(HFMD) were enrolled as controls. Baseline demographic information and vaccination history were collected through parent questionnaires, and blood samples were obtained after informed consent. Neutralizing antibody titers were determined using the standard micro-neutralization assay, and antibody seropositivity rates and GMTs were calculated and statistically compared. The results showed that the vaccination group exhibited a significantly higher antibody seropositivity rate(95.90%) than the control group(62.56%) (χ2 = 65.83, P < 0.0001), with GMTs of 60.46(95% CI: 49.24– 74.24) and 9.49(95% CI: 8.40 – 10.71), respectively(t = 7.032, P < 0.001). Neither sex nor age at first vaccination significantly influenced antibody seropositivity or GMTs. Over time, antibody GMTs declined from 174.9(95% CI: 101.7– 300.5) within six months post-vaccination to 48.03(95%CI: 34.82-66.23) at 2.5 years, with a particularly pronounced decrease after 2.5 years. Despite this decline, the GMTs at 2.5 years remained significantly higher than those in the control group(P < 0.001), indicating sustained immune protection. These findings demonstrate that two doses of the inactivated EV-A71 vaccine elicit high seropositivity rates and robust antibody titers in children, providing effective protection for at least 2.5 years. Given the observed decline in antibody titers, a booster dose administered approximately two to three years after the primary series may be necessary to maintain long-term protective efficacy.
Effect of Jinzhen Oral Liquid on HCoV‑229E and Bacterial Infection‑Induced Pneumonia in Mice
FENG Yixuan;SUN Jing;BAO Lei;GENG Zihan;WANG Xinwei;LI Shuran;ZHANG Jingsheng;XIE Dan;GUO Shanshan;CUI Xiaolan;ZHAO Ronghua;Objective To evaluate the therapeutic effects of Jinzhen Oral Liquid(JZOL) on mouse models of pneumonia induced by human coronavirus 229E(HCoV-229E) and by mixed bacterial infection with Staphylococcus aureus and Streptococcus pneumoniae, and to provide experimental evidence supporting its clinical application. Methods A viral pneumonia model was established by intranasal inoculation of HCoV-229E in mice. Animals were treated with high-, medium-, or low-dose JZOL [30, 15, and 7.5 mL/（kg·d）, corresponding to crude drug doses of 8, 4, and 2 g/（kg·d）, respectively]. Lung index, lung histopathological changes, and viral load in lung tissue were evaluated. Chloroquine phosphate sugar-coated tablets(CQ) and Lianhua Qingwen Granules(LHQW) were used as positive controls. A bacterial pneumonia model was established by intranasal co-infection with Staphylococcus aureus and Streptococcus pneumoniae. The effects of the same JZOL dose regimens on lung index were assessed, with amoxicillin capsules(AMX) and Xiao'er Resuqing Oral Liquid(XRSQ) included as positive controls.Results Treatment with JZOL at all three dose levels reduced the lung index to varying degrees in mice infected with HCoV-229E as well as in those subjected to mixed bacterial infection. In the HCoV-229E model, JZOL significantly decreased viral load in lung tissue and markedly alleviated virus-induced histopathological lung damage.Conclusion These results indicate that JZOL exerts a protective and therapeutic effect against pneumonia induced by HCoV-229E and by co-infection with Staphylococcus aureus and Streptococcus pneumoniae in mice.
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.
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Isolation, Identification and Molecular Evolutionary Characterization of Bovine Herpesvirus 4 from Holstein Friesian in China's mainland
ZHAO Yan;ZHANG Caizhen;WANG Hui;LU Changbin;LI Pinghua;YANG Hang;MA Xueqing;LI Dong;ZHU Xinrong;DU Xiaohua;LU Zengjun;LIU Xia;SUN Pu;Objective To characterize the molecular evolutionary features and biological properties of bovine herpesvirus type 4(BoHV-4) circulating in cattle herds in China, and to evaluate its infectivity and pathogenic potential in a non-natural pregnant host. Methods Vaginal swabs and blood samples were collected from aborted dairy cows on a commercial farm. BoHV-4 and other common bovine pathogens were screened by PCR. BoHV-4 – positive samples were inoculated onto Madin – Darby bovine kidney(MDBK) cells for virus isolation. Viral characteristics were assessed by determining the 50% tissue culture infectious dose(TCID50), constructing multi-step growth curves, and performing phylogenetic analyses based on the glycoprotein B(gB), glycoprotein H(gH), and thymidine kinase(TK) genes. To assess cross-species infectivity, a virus suspension(106.5 TCID50/0.1 mL) was administered to mid-gestation pregnant rabbits via combined intranasal and intramuscular inoculation(0.5 mL per route per animal). In addition, aborted cows were followed for six months to monitor BoHV-4 antibody levels and viral persistence, in order to evaluate the potential long-term effects on reproductive performance. Results BoHV-4 DNA was detected exclusively in vaginal swabs from aborted cows. One BoHV-4 field strain, designated BoHV-4 GSHZ/06/2025 strain, was successfully isolated. The seventh passage reached peak replication at approximately 72 h post-infection in MDBK cells. Phylogenetic analysis indicated that the isolate clustered within Genotype 1 based on the TK gene, but within Genotype 2 based on the gB and gH genes, suggesting a mosaic genomic structure. Recombination analysis further indicated that the gH gene may have arisen from recombination between Genotype 1 and Genotype 2 lineages. Experimental infection showed that combined intranasal and intramuscular inoculation did not establish productive infection in pregnant rabbits. Follow-up results demonstrated that all vaginal swabs and blood samples from 20 aborted cows were negative for BoHV-4 nucleic acid six months later, whereas the seropositivity rate for BoHV-4 antibodies reached 100%. Conclusion The BoHV-4 strain isolated in this study exhibits distinct genetic heterogeneity across genomic regions, likely resulting from recombination events that may influence viral biological properties. The failure to establish infection in a pregnant rabbit model suggests that BoHV-4 may act as an opportunistic or co-infecting pathogen in bovine reproductive disorders. These findings highlight the need to re-evaluate the role of BoHV-4 in cattle reproductive health and provide a foundation for future epidemiological surveillance and control strategies.
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Genotype Replacement and Antigenic Evolution of Hemagglutinin in Highly Pathogenic H5N1 Avian Influenza Viruses
PU Siyu;CUI Simin;LUO Junhao;HE Peiqing;ZHU Haijun;GAO Rongbao;Objective To investigate hemagglutinin(HA) genotype replacement in highly pathogenic H5N1 avian influenza viruses and to characterize their molecular evolutionary trajectories during the 2024 – 2025 outbreak in U.S. dairy farms. In addition, this study aims to elucidate evolutionary patterns at key amino acid residues within antigenic epitopes and their impact on the antigenic landscape. Methods High-quality HA gene sequences of highly pathogenic avian influenza virus(HPAIV) H5N1 were retrieved from the GISAID database. Phylogenetic analysis, positive selection analysis, and frequency profiling of mutations at key amino acid sites were performed to characterize genetic evolution and antigenic variation. In parallel, linear epitope–based antigenicity assessment was conducted to evaluate the structural and functional implications of these key mutations. Results Overall, the H5N1 HA gene exhibited temporal continuity in phylogenetic topology. Strains within the clade 2.3.4.4b lineage, located on peripheral branches, clustered closely with early ancestral lineages, indicating a pattern of evolutionary stasis. In contrast, the clade 2.3.4.4h lineage, despite greater genetic divergence, displayed antigenic reversion through back mutations at positions 131 and 139. During the 2024– 2025 outbreak in U.S. dairy farms, a marked genotype shift occurred from the initially dominant B3.13 genotype to the D1.1 genotype. The B3.13 genotype exhibited a typical pattern of continuous antigenic drift, with residue 147—located in the critical antigenic 130-loop—under positive selection(P < 0.1). Antigenicity prediction suggested that this substitution alters local antigenic index scores and may affect epitope properties, indicating a potential role in immune escape and host adaptation. In contrast, the amino acid constellation at key antigenic sites(positions 131, 211, and 226) in the D1.1 genotype converged toward that of the early North American ancestral strain HA21-NL(2021). This pattern reflects a nonlinear decoupling between genetic distance and antigenic phenotype, resembling the antigenic evolution previously described for the clade 2.3.4.4h lineage. Conclusion H5N1 viruses appear to adopt a complex dual evolutionary strategy in novel mammalian hosts: either achieving adaptive antigenic drift through key mutations(e. g., V147M) or recapitulating mechanisms similar to those observed in the 2.3.4.4h lineage to revert toward earlier antigenic phenotypes. These evolutionary dynamics reshape the antigenic landscape of circulating strains and provide important insights for vaccine strain selection and pandemic risk assessment.
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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.
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Genetic Characterization of a Rabies Virus Isolate from a Human Case in Hainan, China, 2024
ZHANG Minghui;LIN Xiaozheng;XU Xiaonuo;XIE Yonghui;YU Pengcheng;LAO Shijun;LIU Shuqing;CUI Lei;LIU Qian;SUN Chuyang;ZHANG Ling;TAO Xiaoyan;ZHU Wuyang;Objective To determine the transmission source of a human rabies case reported in Changjiang County, Hainan Province, in August 2024 through laboratory testing and genomic analysis. Methods Saliva, cerebrospinal fluid, and whole blood samples from the patient, as well as saliva and brain tissue samples from the biting dog, were collected. Rabies virus(RABV) RNA and virus-neutralizing antibodies were detected using polymerase chain reaction(PCR) and the rapid fluorescent focus inhibition test(RFFIT), respectively. Viral sequences were assembled and aligned using Geneious Prime and MAFFT, and phylogenetic trees were constructed using the maximum likelihood method. Results The patient tested positive for RABV, and the infecting strain was classified within the China I lineage. The nucleoprotein(N) gene sequence was identical to that of a RABV strain(HI25D01) obtained from a dog brain sample collected in March 2025 in Qicha Town, the same locality where the patient resided. Phylogenetic analysis showed that the isolate clustered with Guangxi strains(GXBH2021, GXBH2020, and GXLZ2022) within the same lineage and shared 99.78% nucleotide identity with strain GXBH2021. Conclusion The results indicate that the infecting strain has the potential to establish a stable cycle within the local dog population, posing an ongoing transmission risk. In addition, a potential epidemiological linkage between rabies transmission in Hainan and neighboring Guangxi is suggested, highlighting the need for strengthened regional surveillance and control measures.
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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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