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Genomic Characterization of Echovirus 3 in China
ZHU Chenglin;ZHAO Xiaohong;ZHOU Lei;LIN Jie;XIAO Jinbo;ZHU Shuangli;JI Tianjiao;WANG Dongyan;YANG Qian;SU Fei;LIU Ruifang;YANG Jianfang;YANG Lan;XIAO Kaitao;SHAO Kexin;GAO Shang;XU Yabei;LI Xiaomei;YAN Dongmei;ZHANG Yong;Objective To reveal the molecular evolutionary characteristics and epidemiological patterns of Echovirus 3(E3), improve its genotyping method, elucidate its molecular epidemiological features in China and globally, and provide scientific evidence for the development of E3 prevention and control strategies.Methods Based on 30 E3 strains collected in our laboratory from 2003 to 2023, the full-length VP1 region was amplified and sequenced. Combined with 99 global E3 VP1 full-length sequences from GenBank(45 Chinese sequences and 54 foreign sequences), phylogenetic analysis was performed. BEAST X software was used to estimate the evolutionary rate, time to the most recent common ancestor(tMRCA), and effective population size. Representative strains were subjected to whole-genome sequencing and recombination analysis.Results The VP1 nucleotide similarity between 75 Chinese E3 isolates and the prototype strain Morrisey ranged from 79.7% to 82.8%. Phylogenetic analysis classified global E3 into five genotypes A – E, among which genotypes C and D could be further subdivided into subtypes C1 – C6 and D1 – D4, respectively. Genotype C was the current dominant genotype globally, and the dominant genotype in China was consistent with the global pattern. The average evolutionary rate of the global E3 VP1 region was 2.95 × 10 ⁻ 3 substitutions per site per year, with the most recent common ancestor traced back to 1944. The origin of Chinese E3 isolates was later, with tMRCA of 1981. Global E3 experienced genotype replacement from genotype A to co-circulation of genotypes B, C, and D, and then to predominant circulation of genotype C. E3 circulating in China originated in 1981 and underwent replacement from genotype B to genotype C. Analysis of amino acid mutation sites in the VP1 region revealed genotype-specific mutation sites, providing clues for the replacement of genotype B by genotype C. Whole-genome recombination analysis suggested that E3 underwent recombination with other serotypes of Enterovirus B group in the non-structural protein regions(P2, P3).Conclusion This study improved the global E3 genotyping method based on full-length VP1, revealed the molecular epidemiological characteristics of E3 in China and globally, clarified its evolutionary history and genotype replacement patterns, identified genotype-specific mutation sites, confirmed recombination events in E3, and provided key evidence for the development of prevention and control strategies.
Genomic Characterization and Replication Capacity Analysis of Echovirus 18 in Beijing,China
HAN Zhenzhi;JIA Liping;LIN Chenbo;ZHU Runan;HUANG Hui;DENG Li;ZHAO Linqing;Objective To investigate the genomic features, evolutionary characteristics, and in vitro infection properties of echovirus 18(E18) circulating in Beijing, China. Methods Throat swab specimens were retrospectively collected from pediatric patients diagnosed with hand, foot, and mouth disease(HFMD), herpangina, or related illnesses at the Capital Institute of Pediatrics, Capital Medical University, between March 2010 and October 2019. Samples were screened for pan-enterovirus(pan-EV), EV-A71, coxsackievirus A16(CVA16), CVA6, and CVA10 using real-time reverse transcription PCR(rRT-PCR). Pan-EV-positive samples that could not be serotyped were further subjected to RT-PCR amplification of the VP1 coding region followed by sequencing for genotyping. E18-positive samples were inoculated into RD cells for virus isolation. Full-length genome sequences were obtained by metagenomic sequencing to determine genotypes. Recombination events were analyzed using multiple approaches, including RDP4, SimPlot, and phylogenetic analysis. Viral growth kinetics were assessed by rRT-PCR to compare replication characteristics of a representative strain in RD and Hep-2 cells.Results Among 7,652 clinical specimens, 17 were positive for E18(0.20%, 17/7,652). Fourteen E18 isolates were successfully obtained in RD cells. VP1 sequence analysis demonstrated that all isolates belonged to the C2 subgenotype and clustered with previously reported strains circulating in China. Using strain s6868 as a representative, the complete genome was 7,412 nt in length and shared 80.30% nucleotide identity with the prototype strain Metcalf. Recombination analysis using seven algorithms consistently indicated a potential recombination event with coxsackievirus B5(CVB5) within the P3 coding region(nt 4,006 – 6,592). Replication kinetics showed that in RD cells, E18 RNA copy numbers increased from lg 6.5934 ± 0.1714 copies/μL at 12 h post-infection to lg 8.1852 ± 0.2712 copies/μL, reaching a plateau at 48 h(lg 9.3356 ± 0.3829 copies/μL), which were markedly higher than those observed in Hep-2 cells at 12 h post-infection(lg 5.7121 ± 0.1585 copies/μL). Conclusion E18 strains circulating in Beijing from 2010 to 2019 predominantly belonged to the C2 subgenotype, and the representative strain s6868 may have undergone recombination with CVB5 in the P3 coding region. E18 exhibits more efficient replication in RD cells than in Hep-2 cells, indicating that RD cells are a suitable in vitro model for E18 infection studies.
Correlation Between VP1 Mutations and Replication Dynamics During Mouse Adaptation of Coxsackievirus A16
LI Huijie;WANG Rui;LI Jichen;DUAN Wei;LIANG Yucai;SUN Qiang;ZHANG Yong;National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases (NITFID);Objective To elucidate the molecular basis underlying virulence enhancement of coxsackievirus A16(CVA16) during cross-species adaptation and to establish a phenotypic and genetic evolutionary framework from the parental strain(P0) to the mouse-adapted strain(P5), thereby providing insights into the mechanisms of host adaptation.Methods A clinically isolated CVA16 parental strain(P0) was subjected to serial passages in ICR neonatal mice to generate a mouse-adapted lineage, establishing a complete evolutionary series from P0 to P5. Whole-genome sequencing was performed to compare genetic variations between P0 and P5. RD cell infection assays and neonatal mouse challenge experiments were conducted to systematically evaluate in vitro replication capacity, in vivo virulence, and target-organ histopathological changes across different viral passages. Viral binding and internalization assays were further employed to investigate the molecular mechanisms underlying CVA16 virulence enhancement. Results Comparative genomic analysis identified distinct genetic variations between P0 and P5, including a critical mutation in the VP1 region located within a functional domain associated with viral entry. Phenotypic characterization revealed that the P5 strain exhibited significantly enhanced replication capacity and virulence compared with the parental P0 strain. In RD cells, P5 produced higher levels of viral RNA and increased yields of infectious virions. In neonatal mice, P5 infection resulted in rapid body weight loss, hind-limb paralysis, and 100% mortality. Moreover, viral titers were markedly elevated in skeletal muscle, lung, and spinal cord tissues, accompanied by more severe inflammatory infiltration and structural tissue damage. Mechanistic analyses demonstrated that virulence enhancement of the P5 strain was primarily associated with increased viral internalization efficiency and subsequent intracellular replication, whereas no significant difference was observed in initial virus– cell attachment compared with P0.Conclusion This study demonstrates that virulence enhancement of CVA16 during mouse adaptation is driven by multilayered genetic variations, with increased viral internalization and intracellular replication serving as key determinants. These findings provide experimental evidence for understanding CVA16 cross-species adaptation and neuropathogenesis and establish a foundation for the development of antiviral agents and vaccines targeting structural and functional domains of the viral capsid.
A Study on the Pathogenicity Differences of Six CVA6 Strains with Different Genotypes in 1-day-old ICR Suckling Mice
ZHAO Yingying;WANG Rui;DUAN Wei;LI Jichen;LIANG Yucai;HAN Xiaomei;SUN Qiang;LIU Zhijun;ZHANG Yong;Objective In recent years, Coxsackievirus A6(CVA6) has emerged as the predominant serotype causing hand, foot, and mouth disease(HFMD) in China. Since 2013, the dominant circulating genotype of CVA6 has shifted from the D2 sub-genotype to the D3a sub-genotype. This study aimed to compare the neuropathogenicity of these two sub-genotypes in suckling mice and to explore potential molecular determinants underlying their pathogenic differences, thereby providing a scientific basis for disease control and vaccine development. Methods Two representative CVA6 strains of the D2 sub-genoype and four strains of the D3a sub-genotype were selected. One-day-old ICR suckling mice were infected via intramuscular injection with a dose of 106 50% tissue culture infectious doses(TCID50). Survival rates, body weight changes, and clinical scores were monitored. Viral titers in brain tissues were determined, and neuropathological changes were evaluated by histopathological examination and immunohistochemical analysis. Amino acid sequences of the P1 coding region were compared, and protein structural changes associated with key amino acid substitutions were predicted. Results The peak viral titers of CVA6 D2 and D3a sub-genotypes in suckling mouse brains reached 103.875 TCID50 and 102.25 TCID50, respectively, with a statistically significant difference(P < 0.05). In the 233/HeB/CHN/2011 strain(D2 sub-genotype) infection group, brain tissues exhibited neuronal degeneration and necrosis, loss of Nissl bodies, and pronounced inflammatory responses, accompanied by positive immunohistochemical staining for the VP2 antigen. In contrast, no apparent histopathological lesions or detectable viral antigens were observed in the 26/YC/CHN/2023 strain(D3a sub-genotype) infection group. Amino acid sequence alignment revealed isoleucine(I)/valine(V) substitutions at several key positions, including VP2-172, VP3-49, and VP1-242. Protein structure prediction following single-site amino acid substitutions indicated that substitutions at VP1-54 and VP1-90 altered the hydrogen-bonding network and surface charge distribution within the VP1 loop region, which may affect capsid stability, receptor binding affinity, and viral pathogenicity. Conclusion This study demonstrates marked differences in neuropathogenicity between CVA6 strains of the D2 and D3a sub-genotypes. Through protein structure prediction, key amino acid residues potentially contributing to CVA6 pathogenicity were identified. These sites may serve as important molecular markers for CVA6 molecular epidemiological surveillance and epidemic risk assessment.
Enterovirus Surveillance among Healthy Individuals in Xizang Autonomous Region, China, 2023–2024: Increasing Circulation of Coxsackievirus B
SHAO Kexin;HONG Mei;XIAO Mengyi;ZHOU Lei;YANG Lan;ZHAO Yingying;XIAO Kaitao;ZHU Chenglin;LIN Jie;ZHANG Yong;XING Weijia;XIAO Jinbo;Objective This study aimed to systematically assess the epidemiological characteristics of enteroviruses(EV) among healthy children in the Xizang Autonomous Region,China during 2023 – 2024, providing a basis for regional viral surveillance and disease prevention.Methods A total of 456 fecal samples from healthy children were analyzed, and EV screening was performed using real-time reverse-transcription polymerase chain reaction(real-time RT-PCR). The VP1 region of EV-positive samples was amplified and sequenced by RT-PCR, followed by molecular typing and phylogenetic analysis of the obtained sequences.Results The overall EV detection rate was 21.71%(99/456). Detected viruses were classified into Enterovirus A(EV-A), Enterovirus B(EV-B), and Enterovirus C(EV-C), among which EV-B predominated, accounting for(54.55%, 54/99) of positive samples. The most frequently detected serotypes were Coxsackievirus A4(CVA4, 26/99, 26.26%), Coxsackievirus B5(CVB5, 23/99, 23.23%), Coxsackievirus B4(CVB4, 14/99, 14.14%), Coxsackievirus B2(CVB2, 10/99, 10.10%), and poliovirus type 3(PV3, 9/99, 9.09%). EV infections were mainly observed in children aged 3 – 5 years, with no statistically significant difference between sexes. Regional analysis indicated that Lhasa exhibited the greatest diversity of EV serotypes, while CVA4 and CVB serotypes predominated in Shigatse and Ngari, and CVB5 was most prevalent in Shannan. Phylogenetic analysis showed that CVB2, CVB4, and CVB5 isolates from the Xizang Autonomous Region all belonged to genotype D of their respective serotypes and were closely related to contemporary strains circulating both domestically and internationally. Conclusion This study delineates the epidemiological profile of EVs among healthy children in the Xizang Autonomous Region during 2023– 2024. When combined with surveillance data from 2020 – 2022, a notable increase in the circulation intensity of Coxsackievirus B was observed, underscoring the need for sustained continuous surveillance and targeted prevention and control measures for CVB in this region.
Research Progress on the Promoting Effect of Exosomes in Hepatitis B Virus Transmission,Infection,and Treatment
TANG Xiaoli;WANG Chaofu;WANG Shasha;CHEN Xianglei;CHEN Yujun;CHEN Yunan;LI Lin;Hepatitis B virus( HBV) infection remains a major global public health challenge, with its pathogenic process primarily characterized by hepatocellular injury and inflammatory responses. Exosomes are nanoscale extracellular vesicles secreted by a wide range of cell types, possessing a lipid bilayer membrane and the capacity to transport biologically active molecules, including nucleic acids and proteins, and are widely distributed in body fluids. Through distinct biogenetic and release pathways, exosomes participate in the regulation of diverse physiological and pathological processes, such as immune responses, cell migration, and tumorigenesis. Recent studies have demonstrated that exosomes released from HBV-infected hepatocytes can facilitate hepatitis progression, fibrogenesis, and angiogenesis by transferring viral components and modulating cellular signaling pathways, thereby accelerating disease progression. The pivotal roles of exosomes in viral transmission and host immune regulatory networks are becoming increasingly evident. Further elucidation of the underlying mechanisms is expected to provide valuable insights for innovations in HBV diagnostic approaches, optimization of therapeutic strategies, and the development of novel antiviral agents.
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Correlation Between Viral Load, T-Lymphocyte Exhaustion, and Clinical Prognosis in Gastric Cancer Patients Complicated with Pulmonary Infection
LI Shuai;LI Xuchen;LIU Xianchen;Objective To investigate the association between viral load and the degree of T-lymphocyte exhaustion as well as clinical prognosis in gastric cancer patients complicated with pulmonary infection,and to provide evidence for clinical disease assessment and treatment decision-making.Methods Clinical data from 50 patients admitted between January 2024 and December 2025 were retrospectively analyzed.Based on viral load quantified by real-time fluorescent quantitative PCR,patients were categorized into a high viral load group(n=18,>105 copies/mL),a medium viral load group(n=20,103～105 copies/mL),and a low viral load group(n=12,<103 copies/mL).Baseline characteristics,pulmonary infection-related indicators,inflammatory marker levels,T-lymphocyte exhaustion indicators,and clinical outcomes were compared among the three groups.Oneway analysis of variance(ANOVA) or χ2 tests were used for intergroup comparisons,and pairwise comparisons were performed using the LSD-t test or Bonferroni correction.Results No significant differences were observed among the three groups in baseline characteristics,including gender,age,or gastric cancer TNM stage(all P>0.05).With increasing viral load:(1) Pulmonary infection severity significantly worsened.The rates of severe infection(83.3%) and mechanical ventilation requirement rate(66.7%) in the high viral load group were higher than those in the medium(45.0%,30.0%) and low viral load group(16.7%,8.3%),and the infection control time was significantly prolonged(14.2±3.5 days vs 10.5±2.8 days vs 7.8±2.1 days;P<0.001).(2)Inflammatory marker levels were significantly increased.C-reactive protein(CRP:128.5±25.3 mg/L),procalcitonin(PCT:3.5±1.2 ng/mL),and white blood cell count(WBC:17.8±3.2×109/L) were all significantly higher in the high viral load group than in the medium and low viral load groups(P<0.001).(3)T-lymphocyte exhaustion was more pronounced.Counts of CD3~+T cells(192±45 cells/μL),CD4~+T(98±28 cells/μL),and CD8~+T cells(52±18 cells/μL) were significantly lower in the high viral load group,whereas the proportions of PD-1 positive(40.5±8.2%) and CTLA-4 positive(21.8±5.3%) CD8~+T cells were significantly higher compared with the medium and low viral load groups(P<0.001).(4) Clinical outcomes were poorer.The in-hospital mortality rate(27.8%) and 6-month readmission rate(55.6%) in the high viral load group exceeded those in the medium(10.0%,25.0%) and low viral load group(0.0%,8.3%).Overall survival time(6.5±2.1 months) and progression-free survival time(4.2±1.5 months) were shorter than those in the medium and low viral load groups(P<0.05).Conclusion Elevated viral load is closely associated with aggravated T-lymphocyte exhaustion and poorer clinical outcomes in gastric cancer patients complicated with pulmonary infection.Viral load may serve as an important indicator for assessing disease severity and predicting prognosis in this patient population.
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Research Progress on Coxsackievirus B3 as an Oncolytic Virus
WANG Yulin;MIAO Lei;BAO Wenhao;CHEN Long;WANG Tianying;ZHOU Shaofei;Coxsackievirus B3(CVB3), a member of group B coxsackieviruses, has emerged as a promising oncolytic virus, with its oncolytic potential systematically demonstrated in a variety of solid tumor models. CVB3 exhibits significant oncolytic activity against multiple malignancies, including lung adenocarcinoma, colorectal cancer, breast cancer, endometrial cancer, and pancreatic cancer, through mechanisms such as direct tumor cell lysis, induction of tumor cell apoptosis, activation of anti-tumor immune responses, and remodeling of the tumor microenvironment. Advances in genetic engineering-based targeted modification, directed evolution, and the integration of CVB3 with novel delivery systems have laid a solid experimental foundation for its potential clinical application in cancer therapy. This review summarizes recent progress in the study of CVB3, focusing on its biological characteristics, oncolytic mechanisms, optimization strategies, and synergistic effects in combination with conventional anticancer therapies.
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Dual Nuclear Localization Signals Act Synergistically to Promote Nuclear Import of Duck Plague Virus UL6 Protein
YANG Jing;SONG Di;LIU Sijia;WANG Mingshu;CHENG Anchun;YANG Qiao;Objective Duck plague virus(DPV) is a member of the Alphaherpesvirinae subfamily. The UL6 protein encodes by DPV is a homolog of the herpesviral portal protein and participates in viral capsid assembly and genome packaging within the nucleus. Proper subcellular localization of UL6 is therefore essential for its biological function. This study aims to systematically investigate the subcellular localization of DPV UL6 protein, identify its potential nuclear localization signals and key amino acid residues, and elucidate the molecular mechanism underlying UL6 nuclear import. Methods Indirect immunofluorescence assays were employed to analyze the subcellular localization of the DPV UL6 protein and its mutant forms by detecting Flagtagged UL6 and corresponding mutants expressed in cells. Results Two classical nuclear localization signal motifs were identified within DPV UL6 protein: a bipartite nuclear localization signal(bNLS) and a monopartite nuclear localization signal(mNLS). These two motifs act synergistically to mediate the nuclear import of UL6. Further site-directed mutagenesis revealed that the UL6 K142A, UL6 K737A, UL6 R738A, and UL6 R740A mutants exhibited impaired nuclear localization.Conclusion Both the bNLS and mNLS are essential for the nuclear localization of the DPV UL6 protein, and their synergistic action is required for efficient nuclear import. In particular, residues R738 and R740 are indispensable for UL6 nuclear targeting.
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Association Between Cytomegalovirus Reactivation and Acute Exacerbation of Idiopathic Pulmonary Fibrosis and Its Impact on Prognosis
ZHANG Rui;CHEN Yafei;LI Xiaoli;MING Huaikun;Objective To investigate the association between cytomegalovirus(CMV) reactivation and acute exacerbation of idiopathic pulmonary fibrosis(AE-IPF), and to evaluate the impact of CMV reactivation on the prognosis of patients with IPF.Methods A total of 256 patients with idiopathic pulmonary fibrosis admitted to our hospital between May 2020 and May 2023 were enrolled in this study. According to CMV reactivation status at baseline and during follow-up, patients were divided into a CMV reactivation group(n = 77) and a non-CMV reactivation group(n = 179). Patients were followed up in the outpatient clinic every 3 months, with follow-up continuing until May 2025. Clinical data were collected, CMV DNA levels were monitored during follow-up, and the occurrence of AE-IPF was recorded. Results The proportion of patients receiving glucocorticoid therapy was significantly higher in the CMV reactivation group than in the non-CMV reactivation group(45.45% vs. 28.49%, P < 0.05). During follow-up, AE-IPF occurred in 30 patients(38.96%) in the CMV reactivation group and in 21 patients(11.73%) in the non-CMV reactivation group. Compared with patients without CMV reactivation, those with CMV reactivation had a significantly increased risk of developing AE-IPF(P<0.05). Multivariate analysis identified CMV reactivation and glucocorticoid use as independent risk factors for AE-IPF(P<0.05). A total of 38(49.4%) in the CMV reactivation group and 51 patients(28.5%) in the non-CMV reactivation group died during follow-up. Mortality was significantly higher in the CMV reactivation group(P<0.05). CMV reactivation and the occurrence of AE-IPF were both independent risk factors for poor prognosis in patients with IPF(P<0.05). Conclusion CMV reactivation is an independent risk factor for the development of AE-IPF and for poor prognosis in patients with idiopathic pulmonary fibrosis. In clinical practice, increased vigilance for CMV reactivation is warranted in IPF patients, particularly those receiving glucocorticoid therapy, and enhanced monitoring and management should be implemented.
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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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