Objective Patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection frequently develop central nervous system damage,yet the mechanisms driving this pathology remain unclear.This study investi...Objective Patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection frequently develop central nervous system damage,yet the mechanisms driving this pathology remain unclear.This study investigated the primary pathways and key factors underlying brain tissue damage induced by the SARS-CoV-2 beta variant(lineage B.1.351).Methods K18-hACE2 and C57BL/6 mice were intranasally infected with the SARS-CoV-2 beta variant.Viral replication,pathological phenotypes,and brain transcriptomes were analyzed.Gene Ontology(GO)analysis was performed to identify altered pathways.Expression changes of host genes were verified using reverse transcription-quantitative polymerase chain reaction and Western blot.Results Pathological alterations were observed in the lungs of both mouse strains.However,only K18-hACE2 mice exhibited elevated viral RNA loads and infectious titers in the brain at 3 days post-infection,accompanied by neuropathological injury and weight loss.GO analysis of infected K18-hACE2 brain tissue revealed significant dysregulation of genes associated with innate immunity and antiviral defense responses,including type I interferons,pro-inflammatory cytokines,Toll-like receptor signaling components,and interferon-stimulated genes.Neuroinflammation was evident,alongside activation of apoptotic and pyroptotic pathways.Furthermore,altered neural cell marker expression suggested viral-induced neuroglial activation,resulting in caspase 4 and lipocalin 2 release and disruption of neuronal molecular networks.Conclusion These findings elucidate mechanisms of neuropathogenicity associated with the SARS-CoV-2 beta variant and highlight therapeutic targets to mitigate COVID-19-related neurological dysfunction.展开更多
Ectromelia virus(ECTV),a member of the Orthopoxvirus genus,serves as both a causative agent of mousepox and a pivotal surrogate model for studying highly pathogenic orthopoxviruses.Although genomic data on ECTV remain...Ectromelia virus(ECTV),a member of the Orthopoxvirus genus,serves as both a causative agent of mousepox and a pivotal surrogate model for studying highly pathogenic orthopoxviruses.Although genomic data on ECTV remains limited,we report the isolation and characterization of a novel strain,ECTV-C-Tan-GD01,obtained from rodents in Guangdong Province,China.Nanopore sequencing yielded a complete genome(199 annotated genes,including one gene truncated at the C-terminus)with inverted terminal repeats(ITRs)harboring a conserved hairpin structure.Notably,a frameshift-inducing“G”deletion in the EV159 gene resulted in the truncation of a semaphorin-like protein.In vitro assays demonstrated cell-associated viral replication kinetics,with maximum titers achieved earlier in Vero/HeLa cells(72 h)than in BHK-21/CEF cells(84 h).Murine challenge experiments revealed extreme virulence(LD50<1 plaque-forming unit(PFU)via intranasal/footpad routes)and hepatosplenic tropism.Furthermore,ECTV-C-Tan-GD01 exhibited utility in evaluating orthopoxvirus countermeasures:a single dose of vaccinia virus Tiantan(VTT)or non-replicating vaccinia virus Tiantan(NTV)conferred cross-protection,while tecovirimat(ST-246),cidofovir(CDV),and brincidofovir(initially CMX001)significantly reduced viral loads and pathology.This study establishes ECTV-C-Tan-GD01 as a dual-purpose resource for probing orthopoxvirus evolution and advancing therapeutic development.展开更多
The monkeypox virus(MPXV)has triggered a current outbreak globally.Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control.It is a significant challenge but necessary...The monkeypox virus(MPXV)has triggered a current outbreak globally.Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control.It is a significant challenge but necessary to optimize the strategy and application of rapid full-length genome identification and to track variations of MPXV in clinical specimens with low viral loads,as it is one of the DNA viruses with the largest genome and the most AT-biased,and has a significant number of tandem repeats.Here we evaluated the performance of metagenomic and amplicon sequencing techniques,and three sequencing platforms in MPXV genome sequencing based on multiple clinical specimens of five mpox cases in Chinese mainland.We rapidly identified the full-length genome of MPXV with the assembly of accurate tandem repeats in multiple clinical specimens.Amplicon sequencing enables cost-effective and rapid sequencing of clinical specimens to obtain high-quality MPXV genomes.Third-generation sequencing facilitates the assembly of the terminal tandem repeat regions in the monkeypox virus genome and corrects a common misassembly in published sequences.Besides,several intra-host single nucleotide variations were identified in the first imported mpox case.This study offers an evaluation of various strategies aimed at identifying the complete genome of MPXV in clinical specimens.The findings of this study will significantly enhance the surveillance of MPXV.展开更多
Citation:Liu J,Zhu N,Huo W,et al.Advancing virology research with a human brain organoid platform.hLife 2025;3:237–242.The occurrence of viral infections causing central nervous system(CNS)diseases is significant,oft...Citation:Liu J,Zhu N,Huo W,et al.Advancing virology research with a human brain organoid platform.hLife 2025;3:237–242.The occurrence of viral infections causing central nervous system(CNS)diseases is significant,often accompanied by short-term or long-term sequelae and a high mortality rate.Typical clinical manifestations of viral infections that impact the CNS encompass encephalitis,meningitis,myelitis,and seizures[1].Treatments specific to most viral infections are generally limited.展开更多
In 2022,a global outbreak of mpox was anticipated,with several cases reported in non-endemic countries in early May.Given the challenge of distinguishing the mpox virus(MPXV)from other pathogens based solely on sympto...In 2022,a global outbreak of mpox was anticipated,with several cases reported in non-endemic countries in early May.Given the challenge of distinguishing the mpox virus(MPXV)from other pathogens based solely on symptoms,there is an urgent need for prompt and reliable MPXV detection methods.In this study,we developed assays using recombinase-aided amplification(RAA)to identify MPXV and evaluated their applicability with clinical samples.The assays were designed to detect theN4R gene of MPXV.All assays demonstrated detection limits of 1 copy/μL within the reaction system and exhibited no cross-reactivity with ectromelia or the TianTan strain of vaccinia virus,confirming their high specificity.Our established assay provides results in less than 50 min.Furthermore,we evaluated our assay using clinical samples from laboratory-confirmed mpox patients and demonstrated that the RAA-based assay is valuable for diagnosing MPXV infections in field and clinic settings,especially in areas with limited laboratory resources.Overall,three RAA-based nucleic acid assays for MPXV were established,providing a powerful tool for efficient,rapid,and specific detection of MPXV infection.展开更多
Dear Editor,Mpox(formerly known as monkeypox)is a zoonotic disease caused by infection with the monkeypox virus(MPXV).Mpox cases have been sporadic over the past few decades,with outbreaks occurring in only a limited ...Dear Editor,Mpox(formerly known as monkeypox)is a zoonotic disease caused by infection with the monkeypox virus(MPXV).Mpox cases have been sporadic over the past few decades,with outbreaks occurring in only a limited number of countries,primarily as a result of imported cases(El Eid et al.,2022;Tan and Gao,2022).展开更多
Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),a single-stranded positive RNA virus,exhibits a high rate of genome mutation and recombination(1–2).Recombinant strains of this virus could potentially affe...Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),a single-stranded positive RNA virus,exhibits a high rate of genome mutation and recombination(1–2).Recombinant strains of this virus could potentially affect their binding ability to the ACE2 receptor,which may in turn diminish the protection offered by vaccines and neutralizing antibodies.Notably,the Delta and Omicron recombinations have been collectively referred to as Deltacron(3).Furthermore,XBB.1.5,a recombinant strain resulting from the combination of the BA.2 sublineages BA.2.10.1 and BA.2.75,has demonstrated enhanced transmissibility(4).As such,it is imperative to monitor the genome recombination of SARS-CoV-2 in order to provide valuable insights regarding epidemic and transmission trends.展开更多
基金supported by the National Key Research and Development Program of China(2023YFC3041500).
文摘Objective Patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection frequently develop central nervous system damage,yet the mechanisms driving this pathology remain unclear.This study investigated the primary pathways and key factors underlying brain tissue damage induced by the SARS-CoV-2 beta variant(lineage B.1.351).Methods K18-hACE2 and C57BL/6 mice were intranasally infected with the SARS-CoV-2 beta variant.Viral replication,pathological phenotypes,and brain transcriptomes were analyzed.Gene Ontology(GO)analysis was performed to identify altered pathways.Expression changes of host genes were verified using reverse transcription-quantitative polymerase chain reaction and Western blot.Results Pathological alterations were observed in the lungs of both mouse strains.However,only K18-hACE2 mice exhibited elevated viral RNA loads and infectious titers in the brain at 3 days post-infection,accompanied by neuropathological injury and weight loss.GO analysis of infected K18-hACE2 brain tissue revealed significant dysregulation of genes associated with innate immunity and antiviral defense responses,including type I interferons,pro-inflammatory cytokines,Toll-like receptor signaling components,and interferon-stimulated genes.Neuroinflammation was evident,alongside activation of apoptotic and pyroptotic pathways.Furthermore,altered neural cell marker expression suggested viral-induced neuroglial activation,resulting in caspase 4 and lipocalin 2 release and disruption of neuronal molecular networks.Conclusion These findings elucidate mechanisms of neuropathogenicity associated with the SARS-CoV-2 beta variant and highlight therapeutic targets to mitigate COVID-19-related neurological dysfunction.
基金supported by the Natural Science Foundation of Beijing(7254390)the Youth Science Foundation of Chinese Center for Disease Control and Prevention(2024A103)to W.C.C,the National Key ResearchDevelopment Program of China(2022YFC2304100,2023YFD1800405).
文摘Ectromelia virus(ECTV),a member of the Orthopoxvirus genus,serves as both a causative agent of mousepox and a pivotal surrogate model for studying highly pathogenic orthopoxviruses.Although genomic data on ECTV remains limited,we report the isolation and characterization of a novel strain,ECTV-C-Tan-GD01,obtained from rodents in Guangdong Province,China.Nanopore sequencing yielded a complete genome(199 annotated genes,including one gene truncated at the C-terminus)with inverted terminal repeats(ITRs)harboring a conserved hairpin structure.Notably,a frameshift-inducing“G”deletion in the EV159 gene resulted in the truncation of a semaphorin-like protein.In vitro assays demonstrated cell-associated viral replication kinetics,with maximum titers achieved earlier in Vero/HeLa cells(72 h)than in BHK-21/CEF cells(84 h).Murine challenge experiments revealed extreme virulence(LD50<1 plaque-forming unit(PFU)via intranasal/footpad routes)and hepatosplenic tropism.Furthermore,ECTV-C-Tan-GD01 exhibited utility in evaluating orthopoxvirus countermeasures:a single dose of vaccinia virus Tiantan(VTT)or non-replicating vaccinia virus Tiantan(NTV)conferred cross-protection,while tecovirimat(ST-246),cidofovir(CDV),and brincidofovir(initially CMX001)significantly reduced viral loads and pathology.This study establishes ECTV-C-Tan-GD01 as a dual-purpose resource for probing orthopoxvirus evolution and advancing therapeutic development.
基金supported by the National Key Research and Development Program of China(2022YFC2303401,2022YFC2304100,2016YFD0500301,2021YFC0863300)the Beijing Science and Technology Plan(Z211100002521017)the National Natural Science Foundation of China(82241080)。
文摘The monkeypox virus(MPXV)has triggered a current outbreak globally.Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control.It is a significant challenge but necessary to optimize the strategy and application of rapid full-length genome identification and to track variations of MPXV in clinical specimens with low viral loads,as it is one of the DNA viruses with the largest genome and the most AT-biased,and has a significant number of tandem repeats.Here we evaluated the performance of metagenomic and amplicon sequencing techniques,and three sequencing platforms in MPXV genome sequencing based on multiple clinical specimens of five mpox cases in Chinese mainland.We rapidly identified the full-length genome of MPXV with the assembly of accurate tandem repeats in multiple clinical specimens.Amplicon sequencing enables cost-effective and rapid sequencing of clinical specimens to obtain high-quality MPXV genomes.Third-generation sequencing facilitates the assembly of the terminal tandem repeat regions in the monkeypox virus genome and corrects a common misassembly in published sequences.Besides,several intra-host single nucleotide variations were identified in the first imported mpox case.This study offers an evaluation of various strategies aimed at identifying the complete genome of MPXV in clinical specimens.The findings of this study will significantly enhance the surveillance of MPXV.
基金funded by the National Key Research and Development Program of China(2022YFC2304100 to W.T.)Beijing Natural Science Foundation(M23014 to N.Z.).
文摘Citation:Liu J,Zhu N,Huo W,et al.Advancing virology research with a human brain organoid platform.hLife 2025;3:237–242.The occurrence of viral infections causing central nervous system(CNS)diseases is significant,often accompanied by short-term or long-term sequelae and a high mortality rate.Typical clinical manifestations of viral infections that impact the CNS encompass encephalitis,meningitis,myelitis,and seizures[1].Treatments specific to most viral infections are generally limited.
基金supported by the Beijing Natural Science Foundation(7254390)the Youth Science Foundation of the Chinese Center for Disease Control and Prevention(2024A103)the National Key R&D Program of China(2022YFC2303400).
文摘In 2022,a global outbreak of mpox was anticipated,with several cases reported in non-endemic countries in early May.Given the challenge of distinguishing the mpox virus(MPXV)from other pathogens based solely on symptoms,there is an urgent need for prompt and reliable MPXV detection methods.In this study,we developed assays using recombinase-aided amplification(RAA)to identify MPXV and evaluated their applicability with clinical samples.The assays were designed to detect theN4R gene of MPXV.All assays demonstrated detection limits of 1 copy/μL within the reaction system and exhibited no cross-reactivity with ectromelia or the TianTan strain of vaccinia virus,confirming their high specificity.Our established assay provides results in less than 50 min.Furthermore,we evaluated our assay using clinical samples from laboratory-confirmed mpox patients and demonstrated that the RAA-based assay is valuable for diagnosing MPXV infections in field and clinic settings,especially in areas with limited laboratory resources.Overall,three RAA-based nucleic acid assays for MPXV were established,providing a powerful tool for efficient,rapid,and specific detection of MPXV infection.
基金supported in part by the National Key Research and Development Program of China (2022YFC2303401, 2021YFA1201003, 2021YFC2301605, 2022YFC2304100, 2022YFC2304101, 2022YFC0869900)the National Natural Science Foundation of China (82241066)。
文摘Dear Editor,Mpox(formerly known as monkeypox)is a zoonotic disease caused by infection with the monkeypox virus(MPXV).Mpox cases have been sporadic over the past few decades,with outbreaks occurring in only a limited number of countries,primarily as a result of imported cases(El Eid et al.,2022;Tan and Gao,2022).
基金Supported by National Key R&D Program of China(2022YFC230410,2022 YFC2303401,2021YFC2300101 and 2021YFA 1201003).
文摘Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),a single-stranded positive RNA virus,exhibits a high rate of genome mutation and recombination(1–2).Recombinant strains of this virus could potentially affect their binding ability to the ACE2 receptor,which may in turn diminish the protection offered by vaccines and neutralizing antibodies.Notably,the Delta and Omicron recombinations have been collectively referred to as Deltacron(3).Furthermore,XBB.1.5,a recombinant strain resulting from the combination of the BA.2 sublineages BA.2.10.1 and BA.2.75,has demonstrated enhanced transmissibility(4).As such,it is imperative to monitor the genome recombination of SARS-CoV-2 in order to provide valuable insights regarding epidemic and transmission trends.
