The common fig(Ficus carica L.),Moraceae family,is a commonly grown fruit tree in the Mediterranean region.Fig mosaic disease(FMD)poses a substantial threat to Saudi Arabia’s fig-producing economy.A survey was conduc...The common fig(Ficus carica L.),Moraceae family,is a commonly grown fruit tree in the Mediterranean region.Fig mosaic disease(FMD)poses a substantial threat to Saudi Arabia’s fig-producing economy.A survey was conducted during the two seasons of 2021 and 2022 on fig trees,displaying various fig mosaic disease symptoms.A total of 200 fig leaves and fruit samples were collected from various governorates in several Regions of Saudi Arabia including Riyadh,Tabuk,Hail,Qassim,Al-Jouf,Makkah,Jazan,Al-Madinah,Asir,and Northern Borders.These samples were tested by reverse transcription polymerase chain reaction(RT-PCR)with specific pairs of primers to assess the existence of Fig mosaic virus(FMV),Fig leaf mottle-associated virus 1(FLMaV-1),Fig leaf mottle-associated virus 2(FLMaV-2),Fig mild mottle-associated virus(FMMaV)and Fig fleck-associated virus(FFkaV).The results indicate that four viruses were found in mixed infections and tested positive.FMV was detected with a high infection rate of 46%followed by FLMaV-2 with an infection rate of 20%,FMMaV with an infection rate of 16%,and FLMaV-1 with an infection rate of 7%,respectively,while FFkaV was negative in all tested samples.Sequencing and phylogenetic analysis results showed that the FMV isolates shared 90.3%similarity with other FMV isolates,particularly those from Bosnia and Herzegovina(KU198368).While FLMaV-1 showed 92.5%similarity with the reference isolate FLMaV-1(KX397035),the isolates of FLMaV-2 exhibited 94.7%similarity with reference FLMaV-2 isolate(FN687742),and the isolate of FMMaV showed 94%similarity with reference FMMaV isolate(MG242131)based on sequence comparison.According to the RT-PCR results,FMV was effectively identified in all five fig varieties(Al Faiz Yellow,Asali,Brown Turkey,Iraqi,and Kaab Al-Ghazal).Contrarily,none of these varieties had FLMaV-1,FLMaV-2,or FMMaV.This study investigates the occurrence and economic impact of the viruses that infect fig trees in Saudi Arabia.This suggests that FMV is the primary virus infecting these fig varieties and there is no co-infection with the other tested viruses.These findings underscore the urgent need for implementing region-specific management strategies,such as breeding resistant cultivars,enforcing phytosanitary measures to limit virus spread,and prioritizing vector control to mitigate the economic impact of FMD on Saudi Arabia’s fig industry.展开更多
Introduction: Influenza A (Flu A) and B (Flu B) viruses are responsible for severe acute respiratory infections (SARI) worldwide, with a morbidity of 5 million and mortality of 29,000 - 650,000 deaths per year. Influe...Introduction: Influenza A (Flu A) and B (Flu B) viruses are responsible for severe acute respiratory infections (SARI) worldwide, with a morbidity of 5 million and mortality of 29,000 - 650,000 deaths per year. Influenza B viruses are an important cause of respiratory infections in humans, but they tend to be underappreciated due to the predominance of Influenza A. No molecular study on Influenza B has been carried out in the DRC. This study was conducted to document the molecular evolution of the hemagglutinin (HA) gene in the circulating Influenza B strains over the eight consecutive epidemic seasons (from 2015 to 2022). Methods: Samples were collected from outpatient cases suspected of influenza-like illness (ILI) and in all hospitalized patients with SARI from January 2015 to December 2022. Molecular analysis was done to determine influenza type and subtype, and then segments encoding the HA gene of Influenza B viruses were performed. Results: Of 8497 samples collected and tested, 639 (7.5%) were positive for influenza viruses, including 389 (60.8%) for Influenza A viruses and 248 (38,8%) for Influenza B viruses. Of the positive Influenza B samples, 91 were sequenced, including 26 belonging to the B/Yamagata lineage and 65 to the B/Victoria lineage. The HA gene of Influenza B viruses circulating in the DRC showed deletions in the HA1 region. Molecular analysis of Influenza B viruses reflects the genetic diversity of Influenza B/Yam virus clades (Y2, Y3, Y3V1A) alternating with Influenza B/Victoria virus clades (V1A, V1A.3) depending on the year and influenza seasons. The phylogenetic analysis of these Influenza B strains shows compatibility with the corresponding vaccine strains that the WHO had validated for each influenza season. Conclusion: This study underscores the importance of continuous molecular surveillance of Influenza B viruses in the DRC to understand their epidemiology and evolutionary dynamics. Identifying mutations, such as HA deletions, is critical for assessing their impact on transmissibility vaccine efficacy and guiding effective vaccination and control strategies.展开更多
Viral diseases are an important threat to crop yield,as they are responsible for losses greater than US$30 billion annually.Thus,understanding the dynamics of virus propagation within plant cells is essential for devi...Viral diseases are an important threat to crop yield,as they are responsible for losses greater than US$30 billion annually.Thus,understanding the dynamics of virus propagation within plant cells is essential for devising effective control strategies.However,viruses are complex to propagate and quantify.Existing methodologies for viral quantification tend to be expensive and time-consuming.Here,we present a rapid cost-effective approach to quantify viral propagation using an engineered virus expressing a fluorescent reporter.Using a microplate reader,we measured viral protein levels and we validated our findings through comparison by western blot analysis of viral coat protein,the most common approach to quantify viral titer.Our proposed methodology provides a practical and accessible approach to studying virus-host interactions and could contribute to enhancing our understanding of plant virology.展开更多
Adventitious agents,comprising unintentionally introduced microorganisms in the production of biological products,pose a significant challenge in ensuring the safety of gene therapy products.The revised International ...Adventitious agents,comprising unintentionally introduced microorganisms in the production of biological products,pose a significant challenge in ensuring the safety of gene therapy products.The revised International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH)guildline Q5A(R2)from September 2022 highlights the inclusion of viral vector-based gene therapy products in safety discussions,emphasizing controls in material sourcing,testing,and viral clearance[1].Detecting adventitious virus contamination is complex due to the unique characteristics of gene therapy products and the limitations of routine testing methods.The US Food and Drug Administration(FDA)recommends incorporating routine and specific virus detection methods,including those outlined in various pharmacopeias.Existing control methods have limitations,prompting the need for highly sensitive and broad-spectrum detection approaches.Unlike traditional biological products,gene therapy products primarily consist of live viruses,necessitating methods that distinguish between the main virus and adventitious viruses.Current virus detection techniques,such as polymerase chain reaction(PCR),sequencing,mass spectrometry,and DNA microarrays[2e4],have their drawbacks.展开更多
Diel investigations of water environments are one means to holistically understand the dynamics and functional roles of phytoplankton,bacteria and viruses in these ecosystems.They have the potential to substantially i...Diel investigations of water environments are one means to holistically understand the dynamics and functional roles of phytoplankton,bacteria and viruses in these ecosystems.They have the potential to substantially impact carbon(C),nitrogen(N)and phosphorus(P)biogeochemistry through their respective roles.This study characterizes the phytoplankton,bacteria and virus communities and the elemental composition of various C,N and P nutrients flow over three diel cycles in tropical urban lake.Our results show that ratios of C:N:P fluctuated strongly from the lack of dissolved organic phosphorus(DOP)and PO_(4).Specifically,green algae peaked during day time and exudate dissolved organic matter(DOM)that strongly modulate dissolved organic carbon(DOC):DOP ratio to diel DOP limitation.Multiple linear regression and Stella modelling emphasize the roles of viruses together with Synechococcus as important nutrient recyclers of NH_(4)and PO_(4)in nutrients-limited waters.Respective normalised surface PO_(4)and combined surface and bottom NH_(4)concentration selected both viruses and Synechococcus as important drivers.Process model of N and P biogeochemical cycles can achieve 69%and 57%similar to observed concentration of NH_(4)and PO_(4),respectively.A short latent period of 9 hr was calculated,in addition to the calibrated high infectivity of viruses to Synechococcus.Taken together,the rapid turn-over between Synechococcus and viruses has biogeochemical significance,where the rapid recycling of essential nutrients allows for shortcuts in the N and P cycle,supporting a wide range of microbes.展开更多
Oncogenic viruses include both DNA and RNA viruses which contribute to cancer development by disrupting cellular regulation and interfering in the immune responses.These viruses do not directly cause cancer but instea...Oncogenic viruses include both DNA and RNA viruses which contribute to cancer development by disrupting cellular regulation and interfering in the immune responses.These viruses do not directly cause cancer but instead integrate their genetic material into the host genome thus,affecting cell cycle and tumor suppression.This deregulation also leads to impaired immune function and promotes tumor progression by disrupting the removal of infected cells.Generally,innate immunity consists of two important members,including mitochondria and cell deaths,which impact each other as well.Due to the close correlation between viruses,cell death pathways(apoptosis,necroptosis,and pyroptosis),and mitochondria(mitochondrial antiviral-signaling protein and reactive oxygen species generation),targeting these immune system representatives may offer therapeutic strategies to control the progression of oncogenic viral infections.Some previous studies have covered the association of oncogenic viruses with mitochondria and cell death pathways,respectively,but mitochondria and cell death interact with each other,separately,and this interaction may play a role in the progression of cancer induced by oncogenic viruses.Hence,the purpose of this review is to discuss the relationship between cell death,mitochondria,and viral oncogenesis,focusing on the most surveyed oncogenic viruses’mechanisms of action.展开更多
Interstitial hypertension and extracellular matrix(ECM)barriers imposed by cancer-associated fibroblasts(CAFs)at the tumor site significantly impede the retention of intratumorally administered oncolytic viruses(OVs)a...Interstitial hypertension and extracellular matrix(ECM)barriers imposed by cancer-associated fibroblasts(CAFs)at the tumor site significantly impede the retention of intratumorally administered oncolytic viruses(OVs)as well as their efficacy in infecting and eradicating tumor cells.Herein,a stable,controllable,and easily prepared hydrogel was developed for employing a differential release strategy to deliver OVs.The oncolytic herpes simplex virus-2(oH2)particles were loaded within sodium alginate(ALG),together with the small molecule drug PT-100 targeting CAFs.The rapid release of PT-100 functions as an anti-CAFs agent,reducing ECM,and alleviating interstitial pressure at the tumor site.Consequently,the delayed release of oH2 could more effectively invade and eradicate tumor cells while also facilitating enhanced infiltration of immune cells into the tumor microenvironment,thereby establishing an immunologically favorable milieu against tumors.This approach holds significant potential for achieving highly efficient oncolytic virus therapy with minimal toxicity,particularly in tumors rich in stromal components.展开更多
Though oncolytic viruses(OVs)hold significant potential for comprehensive treatment of malignant tumors,their systemic administration faces substantial challenges such as insufficient circulation time,inadequate tumor...Though oncolytic viruses(OVs)hold significant potential for comprehensive treatment of malignant tumors,their systemic administration faces substantial challenges such as insufficient circulation time,inadequate tumor targeting,and spontaneous antiviral immune response of the body,which seriously limits the clinical application of OVs.Herein,we proposed a tumor targeting strategy of tumor cell membrane biomimetic liposomes to encapsulate OVs for intravenous delivery,which enables OVs to target the homotypic tumor lesions and exert their oncolytic effect.On the one hand,this cell membrane biomimetic carrier enhanced the encapsulation of OVs by the hybrid lipid membranes,concealed the viral capsid proteins,and diminished the neutralization and clearance of the virions from the bloodstream.On the other hand,enhanced tumor targeted delivery can be achieved through the utilization of homologous adhesion molecules on the surface of tumor cell membrane.In addition,this strategy also promoted the tumor infiltration of CD4^(+),CD8^(+)T cells mediated by the oncolytic effect of OVs and increased the levels of inflammatory factors such as tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the tumor,thereby effectively enhancing the anti-tumor effect of intravenous administration of OVs.The findings of our study demonstrate that T-L@Ad11 offers a handy and efficient approach for targeting tumors,thereby enhancing the antitumor efficacy of intravenous administration of OVs.展开更多
On January 30,2024,China announced the first human case of H10N5 influenza infection.Prior to this,human cases of H10N7 and H10N8 had been reported.It is now appropriate to re-examine the evolution and future epidemio...On January 30,2024,China announced the first human case of H10N5 influenza infection.Prior to this,human cases of H10N7 and H10N8 had been reported.It is now appropriate to re-examine the evolution and future epidemiological trends of the H10 and N5 subtypes of avian influenza viruses(AIVs).In this study,we analyzed the reassortment characteristics of the first human-derived H10N5 AIV(A/Zhejiang/ZJU01/2023),as well as the evolutionary dynamics of the wild bird-derived H10 and N5 subtypes of AIVs over the past decade.Our findings indicate that the human-derived H10N5 AIV exhibited low pathogenicity.A/bean_goose/Korea/KNU-10/2022(H10N7)and A/mallard/Novosibirsk_region/962k/2018(H12N5)were identified as the potential reassortment parents.The virus has existed since 2022 and several isolations have been reported in Bangladesh.Phylogenetic analysis showed that H10Ny and HxN5 AIVs in China are clustered differently based on the East Asian-Australian(eastern)and Central Asian-Indian(western)migratory flyways.The H10Ny and HxN5 AIV reassortant strains may cause human infections through accidental spillover.It is possible that another center of AIV evolution,mutation,and reassortment may be developing along the migratory flyways in northeastern Asia,distinct from Europe,the Americas,and China's Yangtze River Delta and Pearl River Delta,which should be closely monitored to ensure the safety of the public.展开更多
Background:Qi pi pill(QPP),which contains Renshen,Baizhu,Fuling,Gancao,Chenpi,Shanyao,Lianzi,Shanzha,Liushenqu,Maiya,and Zexie,was recommended for preventing and treating COVID-19 in Shandong Province(China).However,t...Background:Qi pi pill(QPP),which contains Renshen,Baizhu,Fuling,Gancao,Chenpi,Shanyao,Lianzi,Shanzha,Liushenqu,Maiya,and Zexie,was recommended for preventing and treating COVID-19 in Shandong Province(China).However,the mechanism by which QPP treats infectious diseases remains unclear.This study aims to investigate the therapeutic effect of QPP in vitro and on acute influenza infection in mice,exploring its mechanism of action against influenza A virus(IAV).Methods:The in vitro activity of QPP was assessed using dose–response curve analysis and titer reduction assay,and its antiviral mechanism was identified in vitro by realtime polymerase chain reaction(PCR),time-of-addition,and enzymatic assays.The antiviral efficacy of QPP was further evaluated in vivo using BALB/c mice infected with IAV.At the same time,each single Chinese herbal medicine in QPP was evaluated to preliminarily identify those with antiviral effects.Results:In vitro results showed that QPP exhibited a higher potency antiviral effect against both influenza A and B viruses,inhibiting viral RNA replication and release by targeting RNA-dependent RNA polymerase and neuraminidase.Additionally,QPP significantly decreased the expression of inflammatory cytokines in A549 cells.In vivo study revealed that QPP significantly reduced the lung index and viral load in lung tissue of mice infected with IAV.Renshen,Gancao,Zexie,and Lianzi were the Chinese herbal medicines from QPP that showed anti-IAV activity.Conclusion:The antiviral activity of QPP targets IAV replication and release,cytokine modulation in host cells,and provides protection in mice with acute influenza infection.展开更多
Viruses circulating in small mammals possess the potential to infect humans.Tree shrews are a group of small mammals inhabiting widely in forests and plantations,but studies on viruses in tree shrews are quite limited...Viruses circulating in small mammals possess the potential to infect humans.Tree shrews are a group of small mammals inhabiting widely in forests and plantations,but studies on viruses in tree shrews are quite limited.Herein,viral metagenomic sequencing was employed to detect the virome in the tissue and swab samples from seventy-six tree shrews that we collected in Yunnan Province.As the results,genomic fragments belonging to eighteen viral families were identified,thirteen of which contain mammalian viruses.Through polymerase chain reaction(PCR)and Sanger sequencing,twelve complete genomes were determined,including five parvoviruses,three torque teno viruses(TTVs),two adenoviruses,one pneumovirus,and one hepacivirus,together with three partial genomes,including two hepatitis E viruses and one paramyxovirus.Notably,the three TTVs,named TSTTV-HNU1,TSTTV-HNU2,and TSTTV-HNU3,may compose a new genus within the family Anelloviridae.Notably,TSParvoV-HNU5,one of the tree shrew parvoviruses detected,was likely to be a recombination of two murine viruses.Divergence time estimation further revealed the potential cross-species-transmission history of the tree shrew pneumovirus TSPneV-HNU1.Our study provides a comprehensive exploration of viral diversity in wild tree shrews,significantly enhancing our understanding of their roles as natural virus reservoirs.展开更多
The family Hepeviridae has seen an explosive expansion in its host range in recent years,yet the evolutionary trajectory of this zoonotic pathogen remains largely unknown.The emergence of rat hepatitis E virus(HEV)has...The family Hepeviridae has seen an explosive expansion in its host range in recent years,yet the evolutionary trajectory of this zoonotic pathogen remains largely unknown.The emergence of rat hepatitis E virus(HEV)has introduced a new public health threat due to its potential for zoonotic transmission.This study investigated2?464 wild small mammals spanning four animal orders,eight families,21 genera,and 37 species in Yunnan Province,China.Using broadly reactive reverse transcription-polymerase chain reaction(RT-PCR),we systematically screened the presence and prevalence of Orthohepevirus and identified 192 positive specimens from10 species,corresponding to an overall detection rate of7.79%.Next-generation sequencing enabled the recovery of 24 full-length genomic sequences from eight host species,including Bandicota bengalensis,Eothenomys eleusis,and Episoriculus caudatus,representing newly reported host species for Orthohepevirus strains.Phylogenetic and sequence analyses revealed extensive genetic diversity within orthohepeviruses infecting rodents and shrews.Notably,among the identified strains,20 were classified as Rocahepevirus ratti C1,two as C3,and one as Rocahepevirus eothenomi,while the remaining strain exhibited significant divergence,precluding classification.Evolutionary analyses highlighted close associations between orthohepeviruses and their respective host taxa,with distinct phylogenetic clustering patterns observed across different host orders.These findings emphasize the critical roles of co-speciation and cross-species transmission in shaping the evolutionary trajectories of the genera Paslahepevirus and Rocahepevirus.展开更多
Rift Valley fever virus(RVFV)is a high-containment pathogen that causes severe diseases in humans,with no approved therapeutics available.Its classification as a biosafety level 3(BSL-3)agent has limited research and ...Rift Valley fever virus(RVFV)is a high-containment pathogen that causes severe diseases in humans,with no approved therapeutics available.Its classification as a biosafety level 3(BSL-3)agent has limited research and therapeutic development due to safety concerns.In this study,we developed a stable replicon cell line maintaining the replication of L and S genomic segments of RVFV.Single-cycle viral replicon particles(VRPs)could be efficiently packaged through trans-complementation of glycoproteins from different strains,recapitulating authentic viral entry and replication while minimizing biosafety risks.Using this system,we conducted high-throughput screening of a small-molecule compound library and identified CNX-1351 as an antiviral agent for multiple RNA viruses.Mechanistic studies revealed that CNX-1351 inhibits viral replication,potentially by targeting the PI3K-Akt signaling pathway.This single-cycle VRP system provides a valuable tool for studying RVFV biology,host interactions,antiviral and vaccine development under reduced biosafety constraints.展开更多
The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.Th...The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.These similarities have established the tree shrew as a promising experimental model for biomedical research on cancer,infectious diseases,metabolic disorders,and mental health conditions.Herein,we used metatranscriptomic sequencing to analyze plasma,as well as oral and anal swab samples,from 105 healthy asymptomatic tree shrews to identify the presence of potential zoonotic viruses.In total,eight mammalian viruses with complete genomes were identified,belonging to six viral families,including Flaviviridae,Hepeviridae,Parvovirinae,Picornaviridae,Sedoreoviridae,and Spinareoviridae.Notably,the presence of rotavirus was recorded in tree shrews for the first time.Three viruses-hepacivirus 1,parvovirus,and picornavirus-exhibited low genetic similarity(<70%)with previously reported viruses at the whole-genome scale,indicating novelty.Conversely,three other viruses-hepacivirus 2,hepatovirus A and hepevirus-exhibited high similarity(>94%)to known viral strains.Phylogenetic analyses also revealed that the rotavirus and mammalian orthoreovirus identified in this study may be novel reassortants.These findings provide insights into the diverse viral spectrum present in captive Chinese tree shrews,highlighting the necessity for further research into their potential for crossspecies transmission.展开更多
Periodontitis is the inflammation of the supporting structures around the dentition.Several microbial agents,mostly bacteria,have been identified as causative factors for periodontal disease.On the other hand,oral cav...Periodontitis is the inflammation of the supporting structures around the dentition.Several microbial agents,mostly bacteria,have been identified as causative factors for periodontal disease.On the other hand,oral cavity is a rich reservoir for viruses since it contains a wide variety of cell types that can be targeted by viruses.Traditionally,the focus of research about the oral flora has been on bacteria because the most widespread oral diseases,like periodontitis and dental caries,are outcomes of bacterial infection.However,recently and especially after the emergence of coronavirus disease 2019,there is a growing tendency toward including viruses also into the scope of oral microbiome investigations.The global high prevalence of periodontitis and viral infections may point out to a concomitant or synergistic effect between the two.Although the exact nature of the mechanism still is not clearly understood,this could be speculated through the manipulation of the immune system by viruses;hence facilitating the furthermore colonization of the oral tissues by bacteria.This review provides an extensive and detailed update on the role of the most common viruses including herpes family(herpes simplex,varicella-zoster,Epstein-Barr,cytomegalovirus),Human papillomaviruses,Human immunodeficiency virus and severe acute respiratory syndrome coronavirus 2 in the initiation,progression and prognosis of periodontitis.展开更多
Background:The present comprehensive protocol is focused on the detection of pathogenic enteric RNA viruses,explicitly focusing on norovirus genogroup II(GII),astrovirus,rotavirus,Aichi virus,sapovirus,hepatitis A and...Background:The present comprehensive protocol is focused on the detection of pathogenic enteric RNA viruses,explicitly focusing on norovirus genogroup II(GII),astrovirus,rotavirus,Aichi virus,sapovirus,hepatitis A and E viruses in wastewater treatment plants through droplet digital PCR(ddPCR).Enteric viruses are of significant public health concern,as they are the leading cause of diseases like gastroenteritis.Regular monitoring of environmental samples,particularly from wastewater treatment plants,is crucial for early detection and control of these viruses.This research aims to improve the understanding of the prevalence and dynamics of enteric viruses in urban India and will serve as a model for similar studies in other regions.Our protocol's objective is to establish a novel ddPCRbased methodology for the detection and molecular characterization of enteric viruses present in wastewater samples sourced from Bhopal,India.Our assay is capable of accurately quantifying virus concentrations without standard curves,minimizing extensive optimization,and enhancing sensitivity and precision,especially for low-abundance targets.Methods:The study involves fortnightly collecting and analyzing samples from nine wastewater treatment plants over two years,ensuring comprehensive coverage and consistent data.Our study innovatively applies ddPCR to simultaneously detect and quantify enteric viruses in wastewater,a more advanced technique.Additionally,we will employ next-generation sequencing for detailed viral genome identification in samples tested positive for pathogenic viruses.Conclusion:This study will aid in understanding these viruses’genetic diversity and mutation rates,which is crucial for developing tailored intervention strategies.The findings will be instrumental in shaping public health responses and improving epidemiological surveillance,especially in localities heaving sewage networks.展开更多
The recent concurrent emergence of H5N1,H5N6,and H5N8 avian influenza viruses(AIVs)has led to significant avian mortality globally.Since 2020,frequent human-animal interactions have been documented.To gain insight int...The recent concurrent emergence of H5N1,H5N6,and H5N8 avian influenza viruses(AIVs)has led to significant avian mortality globally.Since 2020,frequent human-animal interactions have been documented.To gain insight into the novel H5 subtype AIVs(i.e.,H5N1,H5N6 and H5N8),we collected 6102 samples from various regions of China between January 2021 and September 2022,and identified 41 H5Nx strains.Comparative analyses on the evolution and biological properties of these isolates were conducted.Phylogenetic analysis revealed that the 41 H5Nx strains belonged to clade 2.3.4.4b,with 13 related to H5N1,19 to H5N6,and 9 to H5N8.Analysis based on global 2.3.4.4b viruses showed that all the viruses described in this study were likely originated from H5N8,exhibiting a heterogeneous evolutionary history between H5N1 and H5N6 during 2015–2022 worldwide.H5N1 showed a higher rate of evolution in 2021–2022 and more sites under positive selection pressure in 2015–2022.The antigenic profiles of the novel H5N1 and H5N6 exhibited notable variations.Further hemagglutination inhibition assay suggested that some A(H5N1)viruses may be antigenically distinct from the circulating H5N6 and H5N8 strains.Mammalian challenge assays demonstrated that the H5N8 virus(21GD001_H5N8)displayed the highest pathogenicity in mice,followed by the H5N1 virus(B1557_H5N1)and then the H5N6 virus(220086_H5N6),suggesting a heterogeneous virulence profile of H5 AIVs in the mammalian hosts.Based on the above results,we speculate that A(H5N1)viruses have a higher risk of emergence in the future.Collectively,these findings unveil a new landscape of different evolutionary history and biological characteristics of novel H5 AIVs in clade 2.3.4.4b,contributing to a better understanding of designing more effective strategies for the prevention and control of novel H5 AIVs.展开更多
Senecavirus A(SVA)has a positive-sense,single-stranded RNA genome.Its 5´untranslated region harbors an internal ribosome entry site(IRES),comprising 10 larger or smaller stem-loop structures(including a pseudokno...Senecavirus A(SVA)has a positive-sense,single-stranded RNA genome.Its 5´untranslated region harbors an internal ribosome entry site(IRES),comprising 10 larger or smaller stem-loop structures(including a pseudoknot)that have been demonstrated to be well conserved.However,it is still unclear whether each stem-loop subdomain,such as a single stem or loop,is also highly conserved.To clarify this issue in the present study,a set of 29 SVA cDNA clones were constructed by site-directed mutagenesis(SDM)on the IRES.The SDM-modified scenarios included:(1)stem-formed complementary sequences exchanging with each other;(2)loop transversion;(3)loop transition;and(4)point mutations.All cDNA clones were separately transfected into cells for rescuing viable viruses,whereas only four SVAs of interest could be recovered,and were genetically stable during 20 passages.One progeny grew significantly slower than the other three did.The dual-luciferase reporter assay showed that none of the SDM-modified IRESes significantly inhibited the IRES activity.Our previous study indicated that a single motif from any of the ten stem structures,if completely mutated,would cause the failure of virus recovery.Interestingly,our present study revealed three stem structures,whose individual complementary sequences could exchange with each other to rescue sequence-modifying SVAs.Moreover,one apical loop was demonstrated to have the ability to tolerate its own full-length transition,also having no impact on the recovery of sequence-modifying SVA.The present study suggested that not every stem-loop structure was strictly conserved in its conformation,while the full-length IRES itself was well conserved.This provides a new research direction on interaction between the IRES and many factors.展开更多
African swine fever(ASF)is a highly fatal hemorrhagic disease afecting domestic pigs caused by African swine fever virus(ASFV).Genetic analysis of ASFV isolates to date has identifed 24 geographically related genotype...African swine fever(ASF)is a highly fatal hemorrhagic disease afecting domestic pigs caused by African swine fever virus(ASFV).Genetic analysis of ASFV isolates to date has identifed 24 geographically related genotypes with various subgroups,but only genotype I and II ASFVs have been reported outside Africa.ASFV genotype II and genotype I viruses were reported in China in 2018 and 2021,respectively.In this study,unique and highly conserved noncoding regions were found between MGF_505-9R and MGF_505-10R in the 188 genomes of ASFV genotypes I and II.A pair of primers was designed on the basis of this region.By optimizing the reaction system and conditions,a SYBR Green I fuorescence PCR assay that can distinguish between ASFV genotypes I and II was established,and the sensitivity,reproducibility and specifcity were evaluated.The detection limit was 1 TCID_(50)/0.1 mL for both genotypes,with no cross-reactivity observed with other common pig pathogens.The intra-and interbatch variation coefcients were both less than 1.2%.Clinical sample detection analysis revealed 47 positive cases out of 100,including 3 for genotype I and 44 for genotype II,aligning with results from the WOAH-recommended and national standard methods.The method developed in this study allows for the diferentiation of ASFV genotypes I and II without the need for genome sequencing,ofering a convenient and rapid approach for ASFV detection and genotype identifcation.展开更多
Objective:Our research aims to evaluate the diagnostic accuracy of colposcopy-guided biopsy(CGB)in detecting high-grade cervical lesions and explore how human papilloma virus(HPV)integration status and other factors a...Objective:Our research aims to evaluate the diagnostic accuracy of colposcopy-guided biopsy(CGB)in detecting high-grade cervical lesions and explore how human papilloma virus(HPV)integration status and other factors affect its performance.Methods:A retrospective cohort analysis involving 550 patients was conducted to evaluate whether the HPV integration plays a role in identifying high-grade cervical lesions and cervical cancer.Logistic regression models and area under the curve(AUC)calculations were employed.Results:Our findings revealed that 53.5%of CGB/surgery pairs demonstrated congruent diagnoses,whereas 17.1%showed underestimation and 29.5%overestimation.Furthermore,multivariate logistic regression analysis identified several key predictors for cervical intraepithelial neoplasia(CIN)2+and CIN3+according to surgical pathology.Notably,a CGB confirming CIN2+[odds ratio(OR)=6.0,95%confidence interval(CI):3.9–9.1,P<0.001],high-grade cytology(OR=2.6,95%CI:1.4–4.9,P=0.003),and HPV integration positivity(OR=2.2,95%CI:1.3–3.5,P<0.001)emerged as significant factors for CIN2+.Similarly,for CIN3+identification,CGB confirming CIN2+(OR=5.3,95%CI:3.4–8.3,P<0.001),high-grade cytology(OR=2.6,95%CI:1.5–4.7,P=0.001),and HPV integration positivity(OR=2.0,95%CI:1.3–3.1,P=0.003)were independent predictors.Conclusion:Our study highlights the innovative role of HPV integration testing as a pivotal adjunct to CGB and cytology,offering a comprehensive approach that may enhance the diagnostic precision for high-grade cervical lesions,ultimately achieving more precise management strategies.展开更多
文摘The common fig(Ficus carica L.),Moraceae family,is a commonly grown fruit tree in the Mediterranean region.Fig mosaic disease(FMD)poses a substantial threat to Saudi Arabia’s fig-producing economy.A survey was conducted during the two seasons of 2021 and 2022 on fig trees,displaying various fig mosaic disease symptoms.A total of 200 fig leaves and fruit samples were collected from various governorates in several Regions of Saudi Arabia including Riyadh,Tabuk,Hail,Qassim,Al-Jouf,Makkah,Jazan,Al-Madinah,Asir,and Northern Borders.These samples were tested by reverse transcription polymerase chain reaction(RT-PCR)with specific pairs of primers to assess the existence of Fig mosaic virus(FMV),Fig leaf mottle-associated virus 1(FLMaV-1),Fig leaf mottle-associated virus 2(FLMaV-2),Fig mild mottle-associated virus(FMMaV)and Fig fleck-associated virus(FFkaV).The results indicate that four viruses were found in mixed infections and tested positive.FMV was detected with a high infection rate of 46%followed by FLMaV-2 with an infection rate of 20%,FMMaV with an infection rate of 16%,and FLMaV-1 with an infection rate of 7%,respectively,while FFkaV was negative in all tested samples.Sequencing and phylogenetic analysis results showed that the FMV isolates shared 90.3%similarity with other FMV isolates,particularly those from Bosnia and Herzegovina(KU198368).While FLMaV-1 showed 92.5%similarity with the reference isolate FLMaV-1(KX397035),the isolates of FLMaV-2 exhibited 94.7%similarity with reference FLMaV-2 isolate(FN687742),and the isolate of FMMaV showed 94%similarity with reference FMMaV isolate(MG242131)based on sequence comparison.According to the RT-PCR results,FMV was effectively identified in all five fig varieties(Al Faiz Yellow,Asali,Brown Turkey,Iraqi,and Kaab Al-Ghazal).Contrarily,none of these varieties had FLMaV-1,FLMaV-2,or FMMaV.This study investigates the occurrence and economic impact of the viruses that infect fig trees in Saudi Arabia.This suggests that FMV is the primary virus infecting these fig varieties and there is no co-infection with the other tested viruses.These findings underscore the urgent need for implementing region-specific management strategies,such as breeding resistant cultivars,enforcing phytosanitary measures to limit virus spread,and prioritizing vector control to mitigate the economic impact of FMD on Saudi Arabia’s fig industry.
文摘Introduction: Influenza A (Flu A) and B (Flu B) viruses are responsible for severe acute respiratory infections (SARI) worldwide, with a morbidity of 5 million and mortality of 29,000 - 650,000 deaths per year. Influenza B viruses are an important cause of respiratory infections in humans, but they tend to be underappreciated due to the predominance of Influenza A. No molecular study on Influenza B has been carried out in the DRC. This study was conducted to document the molecular evolution of the hemagglutinin (HA) gene in the circulating Influenza B strains over the eight consecutive epidemic seasons (from 2015 to 2022). Methods: Samples were collected from outpatient cases suspected of influenza-like illness (ILI) and in all hospitalized patients with SARI from January 2015 to December 2022. Molecular analysis was done to determine influenza type and subtype, and then segments encoding the HA gene of Influenza B viruses were performed. Results: Of 8497 samples collected and tested, 639 (7.5%) were positive for influenza viruses, including 389 (60.8%) for Influenza A viruses and 248 (38,8%) for Influenza B viruses. Of the positive Influenza B samples, 91 were sequenced, including 26 belonging to the B/Yamagata lineage and 65 to the B/Victoria lineage. The HA gene of Influenza B viruses circulating in the DRC showed deletions in the HA1 region. Molecular analysis of Influenza B viruses reflects the genetic diversity of Influenza B/Yam virus clades (Y2, Y3, Y3V1A) alternating with Influenza B/Victoria virus clades (V1A, V1A.3) depending on the year and influenza seasons. The phylogenetic analysis of these Influenza B strains shows compatibility with the corresponding vaccine strains that the WHO had validated for each influenza season. Conclusion: This study underscores the importance of continuous molecular surveillance of Influenza B viruses in the DRC to understand their epidemiology and evolutionary dynamics. Identifying mutations, such as HA deletions, is critical for assessing their impact on transmissibility vaccine efficacy and guiding effective vaccination and control strategies.
基金Funding from Natural Sciences and Engineering Research Council of Canada award number RGPIN/4002-2020.
文摘Viral diseases are an important threat to crop yield,as they are responsible for losses greater than US$30 billion annually.Thus,understanding the dynamics of virus propagation within plant cells is essential for devising effective control strategies.However,viruses are complex to propagate and quantify.Existing methodologies for viral quantification tend to be expensive and time-consuming.Here,we present a rapid cost-effective approach to quantify viral propagation using an engineered virus expressing a fluorescent reporter.Using a microplate reader,we measured viral protein levels and we validated our findings through comparison by western blot analysis of viral coat protein,the most common approach to quantify viral titer.Our proposed methodology provides a practical and accessible approach to studying virus-host interactions and could contribute to enhancing our understanding of plant virology.
基金financially supported by Beijing Municipal Science&Technology Commission,China(Grant No.:Z221100007922015)Youth Development Research Foundation of National Institutes for Food and Drug Control,China(Grant No.:2020B1).
文摘Adventitious agents,comprising unintentionally introduced microorganisms in the production of biological products,pose a significant challenge in ensuring the safety of gene therapy products.The revised International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH)guildline Q5A(R2)from September 2022 highlights the inclusion of viral vector-based gene therapy products in safety discussions,emphasizing controls in material sourcing,testing,and viral clearance[1].Detecting adventitious virus contamination is complex due to the unique characteristics of gene therapy products and the limitations of routine testing methods.The US Food and Drug Administration(FDA)recommends incorporating routine and specific virus detection methods,including those outlined in various pharmacopeias.Existing control methods have limitations,prompting the need for highly sensitive and broad-spectrum detection approaches.Unlike traditional biological products,gene therapy products primarily consist of live viruses,necessitating methods that distinguish between the main virus and adventitious viruses.Current virus detection techniques,such as polymerase chain reaction(PCR),sequencing,mass spectrometry,and DNA microarrays[2e4],have their drawbacks.
文摘Diel investigations of water environments are one means to holistically understand the dynamics and functional roles of phytoplankton,bacteria and viruses in these ecosystems.They have the potential to substantially impact carbon(C),nitrogen(N)and phosphorus(P)biogeochemistry through their respective roles.This study characterizes the phytoplankton,bacteria and virus communities and the elemental composition of various C,N and P nutrients flow over three diel cycles in tropical urban lake.Our results show that ratios of C:N:P fluctuated strongly from the lack of dissolved organic phosphorus(DOP)and PO_(4).Specifically,green algae peaked during day time and exudate dissolved organic matter(DOM)that strongly modulate dissolved organic carbon(DOC):DOP ratio to diel DOP limitation.Multiple linear regression and Stella modelling emphasize the roles of viruses together with Synechococcus as important nutrient recyclers of NH_(4)and PO_(4)in nutrients-limited waters.Respective normalised surface PO_(4)and combined surface and bottom NH_(4)concentration selected both viruses and Synechococcus as important drivers.Process model of N and P biogeochemical cycles can achieve 69%and 57%similar to observed concentration of NH_(4)and PO_(4),respectively.A short latent period of 9 hr was calculated,in addition to the calibrated high infectivity of viruses to Synechococcus.Taken together,the rapid turn-over between Synechococcus and viruses has biogeochemical significance,where the rapid recycling of essential nutrients allows for shortcuts in the N and P cycle,supporting a wide range of microbes.
文摘Oncogenic viruses include both DNA and RNA viruses which contribute to cancer development by disrupting cellular regulation and interfering in the immune responses.These viruses do not directly cause cancer but instead integrate their genetic material into the host genome thus,affecting cell cycle and tumor suppression.This deregulation also leads to impaired immune function and promotes tumor progression by disrupting the removal of infected cells.Generally,innate immunity consists of two important members,including mitochondria and cell deaths,which impact each other as well.Due to the close correlation between viruses,cell death pathways(apoptosis,necroptosis,and pyroptosis),and mitochondria(mitochondrial antiviral-signaling protein and reactive oxygen species generation),targeting these immune system representatives may offer therapeutic strategies to control the progression of oncogenic viral infections.Some previous studies have covered the association of oncogenic viruses with mitochondria and cell death pathways,respectively,but mitochondria and cell death interact with each other,separately,and this interaction may play a role in the progression of cancer induced by oncogenic viruses.Hence,the purpose of this review is to discuss the relationship between cell death,mitochondria,and viral oncogenesis,focusing on the most surveyed oncogenic viruses’mechanisms of action.
基金supported by the National Key R&D Program of China(No.2022YFC2403401)the National Natural Science Foundation of China(Nos.82073368,82303766)+2 种基金the Liaoning Revitalization Talents Program(No.XLYC2007071)the China Postdoctoral Science Foundation(No.2023M743908)the Joint Program of Science and Technology Program of Liaoning Province(No.2023JH2/101700094).
文摘Interstitial hypertension and extracellular matrix(ECM)barriers imposed by cancer-associated fibroblasts(CAFs)at the tumor site significantly impede the retention of intratumorally administered oncolytic viruses(OVs)as well as their efficacy in infecting and eradicating tumor cells.Herein,a stable,controllable,and easily prepared hydrogel was developed for employing a differential release strategy to deliver OVs.The oncolytic herpes simplex virus-2(oH2)particles were loaded within sodium alginate(ALG),together with the small molecule drug PT-100 targeting CAFs.The rapid release of PT-100 functions as an anti-CAFs agent,reducing ECM,and alleviating interstitial pressure at the tumor site.Consequently,the delayed release of oH2 could more effectively invade and eradicate tumor cells while also facilitating enhanced infiltration of immune cells into the tumor microenvironment,thereby establishing an immunologically favorable milieu against tumors.This approach holds significant potential for achieving highly efficient oncolytic virus therapy with minimal toxicity,particularly in tumors rich in stromal components.
基金supported by the National Key R&D Program of China(No.2022YFC2403401)the National Natural Science Foundation of China(nos.82073368,82303766)+2 种基金the Liaoning Revitalization Talents Program(No.XLYC2007071)the China Postdoctoral Science Foundation(No.2023M743908)the Joint Program of Science and Technology Program of Liaoning Province(No.2023JH2/101700094).
文摘Though oncolytic viruses(OVs)hold significant potential for comprehensive treatment of malignant tumors,their systemic administration faces substantial challenges such as insufficient circulation time,inadequate tumor targeting,and spontaneous antiviral immune response of the body,which seriously limits the clinical application of OVs.Herein,we proposed a tumor targeting strategy of tumor cell membrane biomimetic liposomes to encapsulate OVs for intravenous delivery,which enables OVs to target the homotypic tumor lesions and exert their oncolytic effect.On the one hand,this cell membrane biomimetic carrier enhanced the encapsulation of OVs by the hybrid lipid membranes,concealed the viral capsid proteins,and diminished the neutralization and clearance of the virions from the bloodstream.On the other hand,enhanced tumor targeted delivery can be achieved through the utilization of homologous adhesion molecules on the surface of tumor cell membrane.In addition,this strategy also promoted the tumor infiltration of CD4^(+),CD8^(+)T cells mediated by the oncolytic effect of OVs and increased the levels of inflammatory factors such as tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the tumor,thereby effectively enhancing the anti-tumor effect of intravenous administration of OVs.The findings of our study demonstrate that T-L@Ad11 offers a handy and efficient approach for targeting tumors,thereby enhancing the antitumor efficacy of intravenous administration of OVs.
基金funded by the National Natural Science Foundation of China(81872673)the Shanghai New Three-year Action Plan for Public Health(GWVI-11.1-03).
文摘On January 30,2024,China announced the first human case of H10N5 influenza infection.Prior to this,human cases of H10N7 and H10N8 had been reported.It is now appropriate to re-examine the evolution and future epidemiological trends of the H10 and N5 subtypes of avian influenza viruses(AIVs).In this study,we analyzed the reassortment characteristics of the first human-derived H10N5 AIV(A/Zhejiang/ZJU01/2023),as well as the evolutionary dynamics of the wild bird-derived H10 and N5 subtypes of AIVs over the past decade.Our findings indicate that the human-derived H10N5 AIV exhibited low pathogenicity.A/bean_goose/Korea/KNU-10/2022(H10N7)and A/mallard/Novosibirsk_region/962k/2018(H12N5)were identified as the potential reassortment parents.The virus has existed since 2022 and several isolations have been reported in Bangladesh.Phylogenetic analysis showed that H10Ny and HxN5 AIVs in China are clustered differently based on the East Asian-Australian(eastern)and Central Asian-Indian(western)migratory flyways.The H10Ny and HxN5 AIV reassortant strains may cause human infections through accidental spillover.It is possible that another center of AIV evolution,mutation,and reassortment may be developing along the migratory flyways in northeastern Asia,distinct from Europe,the Americas,and China's Yangtze River Delta and Pearl River Delta,which should be closely monitored to ensure the safety of the public.
基金Special Emergency research and development of Social Benefiting Technology Program,Qingdao,Grant/Award Number:23-7-8-smjk-3-nshProject for Development of TCM Science and Technology of Shandong Province,Grant/Award Number:M-2022145+1 种基金Major Basic Program of Natural Science Foundation of Shandong Province,Grant/Award Number:ZR2021ZD17Jinan City Funding for University Innovation Teams,Grant/Award Number:2021GXRC028。
文摘Background:Qi pi pill(QPP),which contains Renshen,Baizhu,Fuling,Gancao,Chenpi,Shanyao,Lianzi,Shanzha,Liushenqu,Maiya,and Zexie,was recommended for preventing and treating COVID-19 in Shandong Province(China).However,the mechanism by which QPP treats infectious diseases remains unclear.This study aims to investigate the therapeutic effect of QPP in vitro and on acute influenza infection in mice,exploring its mechanism of action against influenza A virus(IAV).Methods:The in vitro activity of QPP was assessed using dose–response curve analysis and titer reduction assay,and its antiviral mechanism was identified in vitro by realtime polymerase chain reaction(PCR),time-of-addition,and enzymatic assays.The antiviral efficacy of QPP was further evaluated in vivo using BALB/c mice infected with IAV.At the same time,each single Chinese herbal medicine in QPP was evaluated to preliminarily identify those with antiviral effects.Results:In vitro results showed that QPP exhibited a higher potency antiviral effect against both influenza A and B viruses,inhibiting viral RNA replication and release by targeting RNA-dependent RNA polymerase and neuraminidase.Additionally,QPP significantly decreased the expression of inflammatory cytokines in A549 cells.In vivo study revealed that QPP significantly reduced the lung index and viral load in lung tissue of mice infected with IAV.Renshen,Gancao,Zexie,and Lianzi were the Chinese herbal medicines from QPP that showed anti-IAV activity.Conclusion:The antiviral activity of QPP targets IAV replication and release,cytokine modulation in host cells,and provides protection in mice with acute influenza infection.
基金funded by the National Natural Science Foundation of China(No.U2002218)the Science and Technology Innovation Program of Hunan Province(2024RC1028)Hunan University(No.521119400156).
文摘Viruses circulating in small mammals possess the potential to infect humans.Tree shrews are a group of small mammals inhabiting widely in forests and plantations,but studies on viruses in tree shrews are quite limited.Herein,viral metagenomic sequencing was employed to detect the virome in the tissue and swab samples from seventy-six tree shrews that we collected in Yunnan Province.As the results,genomic fragments belonging to eighteen viral families were identified,thirteen of which contain mammalian viruses.Through polymerase chain reaction(PCR)and Sanger sequencing,twelve complete genomes were determined,including five parvoviruses,three torque teno viruses(TTVs),two adenoviruses,one pneumovirus,and one hepacivirus,together with three partial genomes,including two hepatitis E viruses and one paramyxovirus.Notably,the three TTVs,named TSTTV-HNU1,TSTTV-HNU2,and TSTTV-HNU3,may compose a new genus within the family Anelloviridae.Notably,TSParvoV-HNU5,one of the tree shrew parvoviruses detected,was likely to be a recombination of two murine viruses.Divergence time estimation further revealed the potential cross-species-transmission history of the tree shrew pneumovirus TSPneV-HNU1.Our study provides a comprehensive exploration of viral diversity in wild tree shrews,significantly enhancing our understanding of their roles as natural virus reservoirs.
基金supported by the National Natural Science Foundation of China (U2002218,81874274)Yunnan Health Training Project of High Level Talents (L-2017027)+3 种基金Project of Cross-border Control and Quarantine Innovation Group of Zoonosis of Dali University (ZKPY2019302)to Y.Z.ZOpen Project of Yunnan Key Laboratory of Biodiversity Information (BIKF22-02)Youth Innovation Promotion Association of the Chinese Academy of SciencesYunnan Revitalization Talent Support Program Young Talent Project to X.L.Y。
文摘The family Hepeviridae has seen an explosive expansion in its host range in recent years,yet the evolutionary trajectory of this zoonotic pathogen remains largely unknown.The emergence of rat hepatitis E virus(HEV)has introduced a new public health threat due to its potential for zoonotic transmission.This study investigated2?464 wild small mammals spanning four animal orders,eight families,21 genera,and 37 species in Yunnan Province,China.Using broadly reactive reverse transcription-polymerase chain reaction(RT-PCR),we systematically screened the presence and prevalence of Orthohepevirus and identified 192 positive specimens from10 species,corresponding to an overall detection rate of7.79%.Next-generation sequencing enabled the recovery of 24 full-length genomic sequences from eight host species,including Bandicota bengalensis,Eothenomys eleusis,and Episoriculus caudatus,representing newly reported host species for Orthohepevirus strains.Phylogenetic and sequence analyses revealed extensive genetic diversity within orthohepeviruses infecting rodents and shrews.Notably,among the identified strains,20 were classified as Rocahepevirus ratti C1,two as C3,and one as Rocahepevirus eothenomi,while the remaining strain exhibited significant divergence,precluding classification.Evolutionary analyses highlighted close associations between orthohepeviruses and their respective host taxa,with distinct phylogenetic clustering patterns observed across different host orders.These findings emphasize the critical roles of co-speciation and cross-species transmission in shaping the evolutionary trajectories of the genera Paslahepevirus and Rocahepevirus.
基金Grants from the Program of Shanghai Academic Research Leader(22XD1420600)Shanghai Municipal Science and Technology Major Project(ZD2021CY001)+2 种基金National Natural Science Foundation of China(81974305)Shenzhen Medical Research Fund(SMRF No.B2302029)Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(2023-PT310-02)supported this work.
文摘Rift Valley fever virus(RVFV)is a high-containment pathogen that causes severe diseases in humans,with no approved therapeutics available.Its classification as a biosafety level 3(BSL-3)agent has limited research and therapeutic development due to safety concerns.In this study,we developed a stable replicon cell line maintaining the replication of L and S genomic segments of RVFV.Single-cycle viral replicon particles(VRPs)could be efficiently packaged through trans-complementation of glycoproteins from different strains,recapitulating authentic viral entry and replication while minimizing biosafety risks.Using this system,we conducted high-throughput screening of a small-molecule compound library and identified CNX-1351 as an antiviral agent for multiple RNA viruses.Mechanistic studies revealed that CNX-1351 inhibits viral replication,potentially by targeting the PI3K-Akt signaling pathway.This single-cycle VRP system provides a valuable tool for studying RVFV biology,host interactions,antiviral and vaccine development under reduced biosafety constraints.
基金supported by the National Key R&D Program of China (2021YFC2300900,2021YFC2301300)Academic Promotion Programme of Shandong First Medical University (2019QL006)+2 种基金Natural Science Foundation of Shandong Province (ZR2020QH274)Yunnan Key Research and Development Program (202103AQ100001,202102AA310055)Key Program of Chinese Academy of Sciences (KJZD-SW-L11)。
文摘The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.These similarities have established the tree shrew as a promising experimental model for biomedical research on cancer,infectious diseases,metabolic disorders,and mental health conditions.Herein,we used metatranscriptomic sequencing to analyze plasma,as well as oral and anal swab samples,from 105 healthy asymptomatic tree shrews to identify the presence of potential zoonotic viruses.In total,eight mammalian viruses with complete genomes were identified,belonging to six viral families,including Flaviviridae,Hepeviridae,Parvovirinae,Picornaviridae,Sedoreoviridae,and Spinareoviridae.Notably,the presence of rotavirus was recorded in tree shrews for the first time.Three viruses-hepacivirus 1,parvovirus,and picornavirus-exhibited low genetic similarity(<70%)with previously reported viruses at the whole-genome scale,indicating novelty.Conversely,three other viruses-hepacivirus 2,hepatovirus A and hepevirus-exhibited high similarity(>94%)to known viral strains.Phylogenetic analyses also revealed that the rotavirus and mammalian orthoreovirus identified in this study may be novel reassortants.These findings provide insights into the diverse viral spectrum present in captive Chinese tree shrews,highlighting the necessity for further research into their potential for crossspecies transmission.
文摘Periodontitis is the inflammation of the supporting structures around the dentition.Several microbial agents,mostly bacteria,have been identified as causative factors for periodontal disease.On the other hand,oral cavity is a rich reservoir for viruses since it contains a wide variety of cell types that can be targeted by viruses.Traditionally,the focus of research about the oral flora has been on bacteria because the most widespread oral diseases,like periodontitis and dental caries,are outcomes of bacterial infection.However,recently and especially after the emergence of coronavirus disease 2019,there is a growing tendency toward including viruses also into the scope of oral microbiome investigations.The global high prevalence of periodontitis and viral infections may point out to a concomitant or synergistic effect between the two.Although the exact nature of the mechanism still is not clearly understood,this could be speculated through the manipulation of the immune system by viruses;hence facilitating the furthermore colonization of the oral tissues by bacteria.This review provides an extensive and detailed update on the role of the most common viruses including herpes family(herpes simplex,varicella-zoster,Epstein-Barr,cytomegalovirus),Human papillomaviruses,Human immunodeficiency virus and severe acute respiratory syndrome coronavirus 2 in the initiation,progression and prognosis of periodontitis.
文摘Background:The present comprehensive protocol is focused on the detection of pathogenic enteric RNA viruses,explicitly focusing on norovirus genogroup II(GII),astrovirus,rotavirus,Aichi virus,sapovirus,hepatitis A and E viruses in wastewater treatment plants through droplet digital PCR(ddPCR).Enteric viruses are of significant public health concern,as they are the leading cause of diseases like gastroenteritis.Regular monitoring of environmental samples,particularly from wastewater treatment plants,is crucial for early detection and control of these viruses.This research aims to improve the understanding of the prevalence and dynamics of enteric viruses in urban India and will serve as a model for similar studies in other regions.Our protocol's objective is to establish a novel ddPCRbased methodology for the detection and molecular characterization of enteric viruses present in wastewater samples sourced from Bhopal,India.Our assay is capable of accurately quantifying virus concentrations without standard curves,minimizing extensive optimization,and enhancing sensitivity and precision,especially for low-abundance targets.Methods:The study involves fortnightly collecting and analyzing samples from nine wastewater treatment plants over two years,ensuring comprehensive coverage and consistent data.Our study innovatively applies ddPCR to simultaneously detect and quantify enteric viruses in wastewater,a more advanced technique.Additionally,we will employ next-generation sequencing for detailed viral genome identification in samples tested positive for pathogenic viruses.Conclusion:This study will aid in understanding these viruses’genetic diversity and mutation rates,which is crucial for developing tailored intervention strategies.The findings will be instrumental in shaping public health responses and improving epidemiological surveillance,especially in localities heaving sewage networks.
基金supported by the Science and Technology Program of Guangdong Province(2022B1111010004,2021B1212030015)China Agriculture Research System of MOF and MARA(CARS-41)China National Animal Disease Surveillance and Epidemiological Survey Program(2021–2025)(No.202111).
文摘The recent concurrent emergence of H5N1,H5N6,and H5N8 avian influenza viruses(AIVs)has led to significant avian mortality globally.Since 2020,frequent human-animal interactions have been documented.To gain insight into the novel H5 subtype AIVs(i.e.,H5N1,H5N6 and H5N8),we collected 6102 samples from various regions of China between January 2021 and September 2022,and identified 41 H5Nx strains.Comparative analyses on the evolution and biological properties of these isolates were conducted.Phylogenetic analysis revealed that the 41 H5Nx strains belonged to clade 2.3.4.4b,with 13 related to H5N1,19 to H5N6,and 9 to H5N8.Analysis based on global 2.3.4.4b viruses showed that all the viruses described in this study were likely originated from H5N8,exhibiting a heterogeneous evolutionary history between H5N1 and H5N6 during 2015–2022 worldwide.H5N1 showed a higher rate of evolution in 2021–2022 and more sites under positive selection pressure in 2015–2022.The antigenic profiles of the novel H5N1 and H5N6 exhibited notable variations.Further hemagglutination inhibition assay suggested that some A(H5N1)viruses may be antigenically distinct from the circulating H5N6 and H5N8 strains.Mammalian challenge assays demonstrated that the H5N8 virus(21GD001_H5N8)displayed the highest pathogenicity in mice,followed by the H5N1 virus(B1557_H5N1)and then the H5N6 virus(220086_H5N6),suggesting a heterogeneous virulence profile of H5 AIVs in the mammalian hosts.Based on the above results,we speculate that A(H5N1)viruses have a higher risk of emergence in the future.Collectively,these findings unveil a new landscape of different evolutionary history and biological characteristics of novel H5 AIVs in clade 2.3.4.4b,contributing to a better understanding of designing more effective strategies for the prevention and control of novel H5 AIVs.
基金This work was supported by the National Natural Science Found ation of China(32273000)the Qingdao Demonstration Project for People-benefit from Science and Techniques,China(23-2-8-xdny-14nsh and 24-2-8-xdny-4-nsh)+1 种基金the National Program of Undergraduate Innovation and Entrepreneurship,China(202310435039)the Open Project Fund of State Key Laboratory of Microbial Technology,China(M2023-03)。
文摘Senecavirus A(SVA)has a positive-sense,single-stranded RNA genome.Its 5´untranslated region harbors an internal ribosome entry site(IRES),comprising 10 larger or smaller stem-loop structures(including a pseudoknot)that have been demonstrated to be well conserved.However,it is still unclear whether each stem-loop subdomain,such as a single stem or loop,is also highly conserved.To clarify this issue in the present study,a set of 29 SVA cDNA clones were constructed by site-directed mutagenesis(SDM)on the IRES.The SDM-modified scenarios included:(1)stem-formed complementary sequences exchanging with each other;(2)loop transversion;(3)loop transition;and(4)point mutations.All cDNA clones were separately transfected into cells for rescuing viable viruses,whereas only four SVAs of interest could be recovered,and were genetically stable during 20 passages.One progeny grew significantly slower than the other three did.The dual-luciferase reporter assay showed that none of the SDM-modified IRESes significantly inhibited the IRES activity.Our previous study indicated that a single motif from any of the ten stem structures,if completely mutated,would cause the failure of virus recovery.Interestingly,our present study revealed three stem structures,whose individual complementary sequences could exchange with each other to rescue sequence-modifying SVAs.Moreover,one apical loop was demonstrated to have the ability to tolerate its own full-length transition,also having no impact on the recovery of sequence-modifying SVA.The present study suggested that not every stem-loop structure was strictly conserved in its conformation,while the full-length IRES itself was well conserved.This provides a new research direction on interaction between the IRES and many factors.
基金supported by the National Key Research and Development Program of China(Grant No.2023YFF1000901)the Hubei Agricultural Research System(grant number HBHZD-ZB-2020-005)+3 种基金the National Key Research and Development Program of China(grant number 2021YFD1800101-2)Hubei Hongshan Laboratory(No.2022 hszd023)Project 2662023DKPY004supported by the Fundamental Research Funds for the Central Universities.
文摘African swine fever(ASF)is a highly fatal hemorrhagic disease afecting domestic pigs caused by African swine fever virus(ASFV).Genetic analysis of ASFV isolates to date has identifed 24 geographically related genotypes with various subgroups,but only genotype I and II ASFVs have been reported outside Africa.ASFV genotype II and genotype I viruses were reported in China in 2018 and 2021,respectively.In this study,unique and highly conserved noncoding regions were found between MGF_505-9R and MGF_505-10R in the 188 genomes of ASFV genotypes I and II.A pair of primers was designed on the basis of this region.By optimizing the reaction system and conditions,a SYBR Green I fuorescence PCR assay that can distinguish between ASFV genotypes I and II was established,and the sensitivity,reproducibility and specifcity were evaluated.The detection limit was 1 TCID_(50)/0.1 mL for both genotypes,with no cross-reactivity observed with other common pig pathogens.The intra-and interbatch variation coefcients were both less than 1.2%.Clinical sample detection analysis revealed 47 positive cases out of 100,including 3 for genotype I and 44 for genotype II,aligning with results from the WOAH-recommended and national standard methods.The method developed in this study allows for the diferentiation of ASFV genotypes I and II without the need for genome sequencing,ofering a convenient and rapid approach for ASFV detection and genotype identifcation.
文摘Objective:Our research aims to evaluate the diagnostic accuracy of colposcopy-guided biopsy(CGB)in detecting high-grade cervical lesions and explore how human papilloma virus(HPV)integration status and other factors affect its performance.Methods:A retrospective cohort analysis involving 550 patients was conducted to evaluate whether the HPV integration plays a role in identifying high-grade cervical lesions and cervical cancer.Logistic regression models and area under the curve(AUC)calculations were employed.Results:Our findings revealed that 53.5%of CGB/surgery pairs demonstrated congruent diagnoses,whereas 17.1%showed underestimation and 29.5%overestimation.Furthermore,multivariate logistic regression analysis identified several key predictors for cervical intraepithelial neoplasia(CIN)2+and CIN3+according to surgical pathology.Notably,a CGB confirming CIN2+[odds ratio(OR)=6.0,95%confidence interval(CI):3.9–9.1,P<0.001],high-grade cytology(OR=2.6,95%CI:1.4–4.9,P=0.003),and HPV integration positivity(OR=2.2,95%CI:1.3–3.5,P<0.001)emerged as significant factors for CIN2+.Similarly,for CIN3+identification,CGB confirming CIN2+(OR=5.3,95%CI:3.4–8.3,P<0.001),high-grade cytology(OR=2.6,95%CI:1.5–4.7,P=0.001),and HPV integration positivity(OR=2.0,95%CI:1.3–3.1,P=0.003)were independent predictors.Conclusion:Our study highlights the innovative role of HPV integration testing as a pivotal adjunct to CGB and cytology,offering a comprehensive approach that may enhance the diagnostic precision for high-grade cervical lesions,ultimately achieving more precise management strategies.
