Background:Fruits of Indian gooseberry or Amla or Aonla(Phyllanthus emblica)are important ingredients in many Ayurvedic medicines,but little is known about its leaves.Methods:Different extracts and crude alkaloids of ...Background:Fruits of Indian gooseberry or Amla or Aonla(Phyllanthus emblica)are important ingredients in many Ayurvedic medicines,but little is known about its leaves.Methods:Different extracts and crude alkaloids of P.emblica leaves were evaluated for their antimicrobial activity against clinically important microbes using agar well diffusion assay.The antimicrobial activity of methanolic extract(ME)of P.emblica leaves was also compared with similarly prepared methanolic extracts from leaves of Yellow Kaner(Cascabela peruviana),Parijaat or Harsingar(Nyctanthes arbor-tristis),Custard apple(Annona squamosa),Garlic vine(Mansoa alliacea),Shami plant(Prosopis cineraria),Madar(Calotropis gigantea),and Bael(Aegle marmelos).Results:The ME of leaves of P.emblica was the most potent preparation against bacteria and yeast.Of the 338 strains of microbes belonging to 100 species(96 bacteria and four Candida species),the ME of P.emblica inhibited the growth of 300 strains.A total of 84.62%,96.39%,and 100%of 221 G−bacteria,111 G+bacteria,and 6 Candida species strains,respectively,were inhibited by ME of P.emblica leaves at≤36 mg/well.The aqueous extract of P.emblica leaves also inhibited a similar number of bacterial strains,but at higher concentrations,while the ether extract could inhibit only staphylococci.The alkaloid from P.emblica leaves and the ME from leaves of other plants had insignificant antimicrobial activity at similar≤36 mg/well concentration.Conclusion:The study concluded that the ME of P.emblica leaves may be a useful source of a potent,wide-spectrum antimicrobial substance(s).展开更多
Background:Achromobacter xylosoxidans(A.xylosoxidans)subspecies strains are known to be opportunistic environmental inhabitants.Among the two species,A.xylosoxidans ssp.xylosoxidans is more commonly reported cause of ...Background:Achromobacter xylosoxidans(A.xylosoxidans)subspecies strains are known to be opportunistic environmental inhabitants.Among the two species,A.xylosoxidans ssp.xylosoxidans is more commonly reported cause of nosocomial infections colonizing the hospital environment and medical devices,while A.xylosoxidans ssp.denitrificans(AD)strains are widely distributed in the abiotic environment.The present retrospective observational study was aimed at understanding the occurrence of AD infections in the Bareilly region,and to look into the effective herbal and conventional antimicrobial resistance profile of the strains identified at the laboratory.Methods:The present retrospective study analysed Clinical Microbiology laboratory data of Indian veterinary research Institute.The data for the last 14 years(2011-2024)was retrieved,tabulated and analysed using MS Excel program to determine significance of occurrence,and variation in antimicrobial resistance of the strains isolated from different sources usingχ2 and odds ratio analysis.Results:The study revealed that AD was detectable as a potential pathogen not only from environmental samples but also from 51 clinical cases(either as pure culture or mixed infection),and also from healthy humans and animals.The pathogen was most commonly associated with deaths in animals and birds due to septicaemia and was isolated is single pathogen from blood samples.It was also detected as single pathogen from cases of abortions,metritis,and urinary tract infections.However,from cases of haemorrhagic enteritis,diarrhoea,mastitis,wound infections,pyoderma and abscesses,and middle ear infections AD was isolated in association of one or more potentially pathogenic bacteria.Of the 80 isolates in the study,68 had multiple drug resistance,and 21 produced metallo-β-lactamases responsible for resistance against most of theβ-lactam antibiotics,including cephalosporins and carbapenems.The most effective antibiotic was gentamicin,inhibiting 90.67%of the isolates,followed by tigecycline(85.00%),ciprofloxacin(80.77%),piperacillin tazobactam(80.65%);other antibiotics were effective against less than 80%of the isolates.Among the herbal antimicrobials,cinnamaldehyde,cinnamon oil,carvacrol,and ajowan oil inhibited 98.41%,85.07%,85.00%,and 83.75%of the isolates,respectively.Conclusion:The study concluded that in the Bareilly region in India,multiple-drug-resistance AD may be an emerging pathogen prevalent in environment and apparently healthy animals.More studies are warranted to understand the AD strains at molecular level to understand their zoonotic potential and circulation in the environment.展开更多
The Salmonella pathogenicity islands(SPIs) play crucial roles in the progression of Salmonella infection. In this study, we constructed an improved λ Red homologous recombination system to prepare single and triple d...The Salmonella pathogenicity islands(SPIs) play crucial roles in the progression of Salmonella infection. In this study, we constructed an improved λ Red homologous recombination system to prepare single and triple deletion mutants of 3 prominent SPIs(SPI-1, 2, and 3), aiming at the impact of deletion on morphology, carbon source metabolism, adhesion and invasion capacity, in vivo colonization, and immune efficacy in chicks. Our examination revealed that the surface of the single deletion mutants(SM6ΔSPI1, ΔSPI2, and ΔSPI3) exhibited a more rugged texture and appeared to be enveloped in a layer of transparent colloid, whereas the morphology of the triple deletion mutant(SM6ΔSPI1&2&3) remained unaltered when compared to the parent strain. The carbon metabolic spectrum of the SPI mutants underwent profound alterations, with a notable and statistically significant modification observed in 30 out of 95 carbon sources, primarily carbohydrates(17 out of 30). Furthermore, the adhesion capacity of the 4 mutants to Caco-2 cells was significantly reduced when compared to that of the parent strain. Moreover,the invasion capacity of mutants SM6ΔSPI1 and SM6ΔSPI1&2&3 exhibited a substantial decrease, while it was enhanced to varying degrees for SM6ΔSPI3 and SM6ΔSPI2. Importantly, none of the 4 mutants induced any clinical symptoms in the chicks. However, they did transiently colonize the spleen and liver. Notably, the SM6ΔSPI1&2&3mutant was rapidly cleared from both the spleen and liver within 8 days post-infection and no notable pathological changes were observed in the organs. Additionally, when challenged, the mutants immunized groups displayed a significant increase in antibody levels and alterations in the CD3+CD4+ and CD3+CD8+ subpopulations, and the levels of IL-4 and IFN-γ cytokines in the SM6ΔSPI1&2&3 immunized chicken serum surpassed those of other groups.In summary, the successful construction of the 4 SPI mutants lays the groundwork for further exploration into the pathogenic(including metabolic) mechanisms of SPIs and the development of safe and effective live attenuated Salmonella vaccines or carriers.展开更多
The inappropriate use of cephalosporins lead to the occurrence and global spread of bacteria resistant to these antimicrobials.In this study,we isolated four Escherichia albertii strains from broilers in eastern China...The inappropriate use of cephalosporins lead to the occurrence and global spread of bacteria resistant to these antimicrobials.In this study,we isolated four Escherichia albertii strains from broilers in eastern China.The antimicrobial susceptibility and genomic characterization of these E.albertii isolates were determined.Our results revealed that these four E.albertii isolates exhibited resistance to tetracyclines,chloramphenicol,β-lactams,aminoglycosides,polymyxin B,sulfonamides,quinolones,and other antimicrobials.Among them,EA04 isolate was multidrug resistant and harbored extended-spectrumβ-lactamases(ESBL)genes blaCTX-Mand blaTEM.Whole genome sequencing and core-genome multilocus sequence typing(cgMLST)based on all ST4638 E.albertii for EA04 inferred highly probable epidemiological links between selected human isolates.Additionally,the ESBL genes blaTEM-141and blaCTX-M-55were coexistent in an approximately 75 kb Inc FII plasmid pEA04.2 in EA04.Comparative analysis indicated that genes blaTEM-141and blaCTX-M-55were located in IS15-blaCTX-M-55-wbu C-blaTEM-141-IS26 region,which similar structures were identified in various bacteria.Furthermore,the plasmid pEA04.2 could be transferable to E.coli EC600 and lead to the resistance to third-generation cephalosporins.These results suggested that chicken potentially serve as a reservoir for multidrug resistant E.albertii,which increases the risk of horizontal transfer of antimicrobial resistance between humans,animals and environment.展开更多
Salmonella enterica serovar Typhimurium,the causative agent of gastroenteritis,is one of the most successful intracellular pathogens.Although certain host factors for Salmonella infection have been unveiled,the factor...Salmonella enterica serovar Typhimurium,the causative agent of gastroenteritis,is one of the most successful intracellular pathogens.Although certain host factors for Salmonella infection have been unveiled,the factors mediating Salmonella entry,particularly the invasion process,remain obscure.Here,we have unearthed β2 integrin,a crucial member of the integrin family,as an important host factor facilitating Salmonella invasion.It is demonstrated that overexpression of β2 integrin promotes Salmonella invasion,while the knockdown of β2 integrin significantly diminishes the extent of invasion.Moreover,Salmonella exhibits specific binding affinity towards β2 integrin,and the block of β2 integrin on cell surface substantially reduces the infection of cells in vitro.The ectodomain soluble protein of β2 integrin neutralized Salmonella infection both in cells(in vitro)and in mice(in vivo).Additionally,Salmonella protein YrbD directly interacts with β2 integrin to facilitate its invasion.To our knowledge,this study showed for the first time that the protein YrbD mediates Salmonella adhesion and internalization into host cells by interacting with β2 integrin.These findings not only broaden our understanding of the mechanisms underlying Salmonella entry,but also identify a prospective target for therapeutic control.展开更多
As an emerging genotype,the G9 genotype rotaviruses(RVs)are widespread among humans and pigs,and have been reported in many countries and regions in the recent years.Moreover,porcine G9 strains could cross the intersp...As an emerging genotype,the G9 genotype rotaviruses(RVs)are widespread among humans and pigs,and have been reported in many countries and regions in the recent years.Moreover,porcine G9 strains could cross the interspecies barrier to infect human.To investigate the epidemic trends of porcine G9 strains as well as the cross-immunoreactivity among different isolates,an epidemiological investigation about porcine G9 genotype RVs(PoRVs)was performed during the period 2020-2023 in multiple provinces of China.A total of nine representative strains were identified.The phylogenetic analysis based on viral VP7 gene showed that these strains mainly clustered with lineages Ⅲ and Ⅵ,which revealed the predominant G9 PoRVs in China.Moreover,a new lineage,lineage Ⅶ,was identified,and strains of this lineage were found to be circulating in Guangdong and Taiwan.Except lineages Ⅰ and Ⅳ,some isolates from other lineages could co-circulate in pigs and humans.Three G9 strains,namely 923H,923E,and 923X,which belonged to the largest sub-lineage Ⅲ,were isolated.Then,the significant cross-reactivity was observed among strains of the same or different lineages.This study is the first to systematically investigate the genetic and immunogenetic characteristics of porcine G9 genotype rotavirus in China,as well as the potential cross-species transmission between pigs and humans,providing a valuable direction for the effective prevention of porcine rotavirus.展开更多
Dear editor,As of 2023,the domestic cat population in China reached 65 million,surpassing dogs to become the most numerous companion animal in the country.Feline calicivirus(FCV)infection,one of the three most prevale...Dear editor,As of 2023,the domestic cat population in China reached 65 million,surpassing dogs to become the most numerous companion animal in the country.Feline calicivirus(FCV)infection,one of the three most prevalent infectious diseases in cats,poses a severe threat to feline health.FCV,classified under the Caliciviridae family(genus Vesivirus).展开更多
African swine fever(ASF)is an acute,hemorrhagic,and highly contagious disease in pigs caused by the African swine fever virus(ASFV).Our previous studies have demonstrated that deletion of the MGF360-9L gene weakens AS...African swine fever(ASF)is an acute,hemorrhagic,and highly contagious disease in pigs caused by the African swine fever virus(ASFV).Our previous studies have demonstrated that deletion of the MGF360-9L gene weakens ASFV virulence in pigs,yet the underlying mechanism remains unclear.To investigate the mechanism of MGF360-9L regulating ASFV pathogenicity,the relationship between MGF360-9L and host proteins was identified by mass spectrometry.We found that host protein DEAD-box helicase 20(DDX20)interacted with and colocalized with MGF360-9L.Overexpression of DDX20 inhibited ASFV replication,whereas knockdown of DDX20 had the opposite effects.Moreover,DDX20 inhibited ASFV replication by promoting the activation of type I interferon signaling.Surprisingly,DDX20 was gradually degraded following ASFV infection.Mechanistically,MGF360-9L promoted the autophagic degradation of DDX20 by recruiting autophagy-related protein Ras-related protein Rab-1A(Rab1A).Silencing Rab1A suppressed ASFV replication,while overexpression of Rab1A exhibited the opposite effects.Furthermore,Rab1A,MGF360-9L and DDX20 could form a complex to facilitate the degradation of DDX20.Knockdown of Rab1A impaired MGF360-9L-mediated degradation of DDX20 during ASFV infection.In summary,our study demonstrates that MGF360-9L targets DDX20 for autophagy degradation to antagonize its antiviral function and facilitate ASFV replication.This finding broadens our understanding of the regulatory network between ASFV and its host,and provides new insights into the pathogenesis and immune evasion mechanisms of ASFV.展开更多
Background:Although many sugars are known antibacterial in higher concentrations(>50%W/V)and a few at low concentrations too(≤1 mg/mL),most of the studies are limited to only a few reference isolates,therefore no ...Background:Although many sugars are known antibacterial in higher concentrations(>50%W/V)and a few at low concentrations too(≤1 mg/mL),most of the studies are limited to only a few reference isolates,therefore no concrete conclusion can be drawn.Methods:This study evaluated the antimicrobial potential of 19 sugars against 8000 isolates of bacteria belonging to 46 genera and also against 30 reference strains of microbes of 14 different species.To determine susceptibility to sugars,a sugar-disc(1 mg)diffusion assay was performed on Mueller Hinton agar using the same method used for antimicrobial susceptibility of microbial strains.Results:In the study,of 3,336 isolates of Gram-positive bacteria 147(4.35%)were susceptible to one or more sugars but only 16(0.37%)of the 4644 isolates of Gram-negative bacteria were susceptible to one or other sugar.Gram-positive bacteria were significantly more often susceptible to one or more sugars than isolates of Gram-negative bacteria(OR 13.33,CI99,6.74–26.37).A total of 163 test-isolates(2.04%)but none of the reference strains were sugar susceptible.Most of the isolates susceptible to sugars(135 of 2,295)were members of the Bacillaceae(36/679)and Micrococcaceae(99/1,616)family.However,out of 5,705 isolates belonging to other bacterial families,only 28 isolates(0.49%)were susceptible to one or more sugars.The most effective to least effective sugars as antibacterial were mannose,inositol,mannitol,sucrose,raffinose,ribose,xylose,trehalose,dulcitol,maltose,lactose,inulin,salicin,melibiose,sorbitol,adonitol,arabinose,glucose,and esculin,inhibiting 69,58,23,14,11,10,10,7,7,7,6,6,5,5,5,5,2,1,and 1 of the test-isolates,respectively.Conclusion:The results are still intriguing in determining the utility of sugar susceptibility of different bacteria and are still beyond making any conclusion for their therapeutic utility.However,the study can be concluded that Gram-positive bacteria are generally more susceptible to lower concentrations of different sugars than Gram-negative bacteria,and various sugars have variable selectivity in their antibacterial effect on multiple types of bacteria.展开更多
Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on...Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase.Full-length cDNA of the LN16-A strain was constructed by assembling 5 cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme.Transfection of this plasmid,along with the supporting plasmids encoding the N,P,M2-1,and L proteins of LN16-A into BSR-T7/5 cells,resulted in the recovery of aMPV subtype B.To identify an effective insertion site,the enhanced green fluorescent protein(EGFP)gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As.The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites(between the leader and N genes or replacing the SH gene).To verify the availability of the site between G and L for foreign gene expression,the VP2 gene of very virulent infectious bursal disease virus(vvIBDV)was inserted into this site,and recombinant LN16-A(rLN16A-vvVP2)was successfully rescued.Single immunization of specificpathogen-free chickens with rLN16A-vvVP2 induced high levels of neutralizing antibodies and provided 100%protection against the virulent aMPV subtype B and vvIBDV.Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.展开更多
Infectious bursal disease(IBD)is an acute,highly contagious disease that affects chicks(Müller et al.2003).IBD mainly damages the immune organs of chicks,especially the central immune organ,causing immune suppres...Infectious bursal disease(IBD)is an acute,highly contagious disease that affects chicks(Müller et al.2003).IBD mainly damages the immune organs of chicks,especially the central immune organ,causing immune suppression in diseased chicks(Muller et al.2012).The pathogenic infectious bursal disease virus(IBDV)is a member of the Avira virus genus in the Birnaviridae family.(Harkness et al.1975;Dobos et al.1979;Müller et al.1979).IBDV is prevalent worldwide,causing serious economic losses to the global poultry industry.Currently,vaccination remains the most cost-effective way to prevent IBDV.展开更多
Influenza A viruses(IAVs)are single-stranded negative-sense RNA viruses that continually challenge animal and human health.In IAV-infected cells,host RNA-binding proteins play key roles in the life cycle of IAV by dir...Influenza A viruses(IAVs)are single-stranded negative-sense RNA viruses that continually challenge animal and human health.In IAV-infected cells,host RNA-binding proteins play key roles in the life cycle of IAV by directly binding to viral RNA.Here,we examined the role of the host RNA-binding protein nucleophosmin-1(NPM1)in IAV replication.We found that,as a nucleolar phosphoprotein,NPM1 directly binds to viral RNA(vRNA)and inhibits the replication of various subtypes of IAV.NPM1 binding to vRNA competitively reduces the assembly of the viral ribonucleoprotein complex and the viral polymerase activity,thereby reducing the generation of progeny viral RNA and virions.The RNA-binding activity of NPM1,with the key residues T199,T219,T234,and T237,is essential for its anti-influenza function.Taken together,our findings demonstrate that NPM1 acts as an RNA-binding protein and interacts with IAV vRNA to suppress viral replication.展开更多
In this study,we developed a highly sensitive enzyme-linked immunosorbent assay(ELISA)using newly produced monoclonal antibodies(mAbs)for detecting horse/donkey IL-1βin cell culture medium and serum samples.The mAbs ...In this study,we developed a highly sensitive enzyme-linked immunosorbent assay(ELISA)using newly produced monoclonal antibodies(mAbs)for detecting horse/donkey IL-1βin cell culture medium and serum samples.The mAbs were generated via the use of a KLH-conjugated peptide and purified equine IL-1βprotein as separate immunogens.Notably,the generated mAbs(3G8 and 5G3)demonstrated no cross-reactivity with other major inflammatory mediators,including IL-1α,IL-1Ra,TNF-α,and SAA.The IL-1βassay,which is based on the screened mAbs,exhibits a detection range of 200-10,000 pg/mL,meeting clinical detection requirements.The coefficients of variation for the repeatability and reproducibility of the assay were both less than 5%,indicating an acceptable level of variation.Subsequently,84 equine and 24 asinine serum samples were collected,and the IL-1βconcentration was measured with both our assay and a commercial kit in parallel.Our results revealed no significant difference between the in-house and commercial ELISA kits for the detection of IL-1βconcentrations in horse sera.Moreover,our ELISA method demonstrated superior sensitivity for IL-1βdetection in donkey samples compared to existing commercial assays.These findings suggest that the newly developed ELISA provides a reliable analytical method for detecting IL-1βin both equine and asinine samples.展开更多
Objective:The objective of this study was to determine the level of methotrexate(MTX)toxicity in the intestines of mice and to evaluate the protective effect of probiotics composed of Streptococcus,Bifidobacterium,and...Objective:The objective of this study was to determine the level of methotrexate(MTX)toxicity in the intestines of mice and to evaluate the protective effect of probiotics composed of Streptococcus,Bifidobacterium,and Lactobacillus species on intestinal cells during MTX treatment.Methods:Mice were divided into three groups:control,MTX group(received MTX injections),and MTX+probiotics group(received MTX injections along with a diet containing probiotics).Morphological and histological changes,the level of mitochondrial DNA(mtDNA)damage,the level of lipid peroxidation products,and gene expression in the mice’s small intestine were assessed.Results:We demonstrated that intraperitoneal MTX injections significantly increased mtDNA damage in the liver(p<0.001),small intestine(p<0.001),and blood of mice(p<0.01).MTX elevated the quantity of lipid peroxidation products in the liver and small intestine,indicating its strong prooxidative properties.MTX induced structural changes in the mice’s intestines,characterized by leukocytic infiltration of tissues.Probiotic therapy in mice partially mitigated the morphological and histological changes in the small intestine induced by MTX,reduced oxidative stress,and promoted increased expression of quinone oxidoreductase 1(Nqo1),which participates in both cell protection against oxidative stress and drug/xenobiotic detoxification.Probiotics prevented the upregulation of the proinflammatory cytokine IL-1b in the small intestine and induced increased expression of genes associated with the Nuclear factor erythroid 2-related factor 2/Antioxidant response element(Nrf2/ARE)pathway,an important mechanism of cell protection.Conclusions:Probiotics can be considered an effective approach to reducing the toxicity of MTX during psoriasis or cancer treatment.展开更多
Monkeypox virus(MPXV),a member of the Orthopoxvirus genus,caused a large-scale global outbreak in 2022.Developing mouse models for MPXV infection is crucial for advancing research on vaccines and therapeutic intervent...Monkeypox virus(MPXV),a member of the Orthopoxvirus genus,caused a large-scale global outbreak in 2022.Developing mouse models for MPXV infection is crucial for advancing research on vaccines and therapeutic interventions.To address this,we conducted a comparative study on the susceptibility of six mouse strains—severe combined immune-deficiency(SCID),nude,genetically diabetic(db/db)and obese(ob/ob),C57BL/6J,and BALB/c—to MPXV infection.Mouse strains were infected with MPXV via intranasal inoculation,and body weight changes and mortality were monitored post-infection.Additionally,the tissue distribution of MPXV and the pathological changes in the lung tissues of the infected mice were evaluated.The results demonstrated that SCID and nude mice exhibited significant weight loss following MPXV infection,with 100%mortality observed in SCID mice,while no mortality occurred in nude mice.In contrast,the other mouse strains showed no significant weight loss or mortality.Notably,the viral load in the lung tissues of SCID and nude mice was the highest among the tested strains.Furthermore,we investigated the impact of different inoculation routes—intranasal(I.N.),intraperitoneal(I.P.),and intravenous(I.V.)—on the pathogenicity of MPXV in mice.The results revealed that the intravenous route induced more pronounced pathogenic effects compared to the intranasal and intraperitoneal routes.In summary,this study provides valuable insights into the development of MPXV-infected mouse models,offering a foundation for further research on MPXV pathogenesis and therapeutic drug development.展开更多
Avian leukosis is an important tumorigenic disease caused by the avian leukosis virus(ALV)in poultry.ALVs belong to the retroviral family and are classified into 11 subgroups(ALV-A to ALV-K).Among them,ALV-J was first...Avian leukosis is an important tumorigenic disease caused by the avian leukosis virus(ALV)in poultry.ALVs belong to the retroviral family and are classified into 11 subgroups(ALV-A to ALV-K).Among them,ALV-J was first introduced into China in 1999,spreading widely and evolving from infecting meat-type chickens to layer chickens and Chinese local chickens.ALV-J typically induces myeloid leukosis in infected chickens,but also induces a high proportion of hemangiomas in infected layer chickens,posing a serious threat to poultry breeds in China.As a retrovirus,the genome of ALV-J has undergone significant mutations,which may be related to the expansion of the infection host range and increased pathogenicity of ALV-J.Over the last two decades,the introduction and spread of ALV-J in China have caused substantial losses to the poultry industry.Specialized detection assays have been developed to combat ALV-J infections in China.Additionally,ongoing research aims to employ gene-editing technology as a novel antiviral strategy to control the spread of ALV infections.This review highlights the importance of understanding the impact of ALV-J on the Chinese poultry industry and emphasizes the need for ongoing research and innovation to safeguard poultry health and promote sustainable poultry farming practices in China.展开更多
African swine fever(ASF),caused by African swine fever virus(ASFV),is a highly contagious swine disease that has spread globally.Effective control strategies are not yet available.In this study,we prepared K205R mRNA,...African swine fever(ASF),caused by African swine fever virus(ASFV),is a highly contagious swine disease that has spread globally.Effective control strategies are not yet available.In this study,we prepared K205R mRNA,which was then formulated using Lipid Nanoparticle(LNP).The resulting K205R mRNA-LNP showed a particle size of approximately 86.27 nm and an mRNA encapsulation efficiency of 96.24%.Efficient expression of the K205R protein was confirmed in both HEK293T and PK15 cells.We further evaluated the immunogenicity of K205R mRNA-LNP in mice and pigs.All immunized animals developed significantly higher levels of IgG antibodies against K205R compared to the control group in the first week after the second immunization,with antibody titers reaching up to 105.Challenge experiments showed that K205R mRNA delayed the time of death.Our results suggested the successful implementation of the mRNA platform in the preparation and application of ASFV mRNA.展开更多
Anoikis is a specialized form of programmed cell death triggered by the detachment of cells from the extracellular matrix(ECM).Tumor cells that develop resistance to anoikis acquire the ability to detach,migrate,and c...Anoikis is a specialized form of programmed cell death triggered by the detachment of cells from the extracellular matrix(ECM).Tumor cells that develop resistance to anoikis acquire the ability to detach,migrate,and colonize distant sites,ultimately leading to the formation of metastatic tumors.Bit1(Bcl-2 inhibitor of transcription 1),a key effector of anoikis,is released into the cytoplasm upon loss of cell attachment and activates a caspase-independent pathway of apoptosis.Newcastle disease virus(NDV),a pathogen that poses a significant threat to the poultry industry,has also emerged as a promising oncolytic virus capable of selectively targeting and killing tumor cells.However,whether NDV can induce the death of anoikis-resistant tumor cells by activating Bit1 remains unclear.In this study,we utilized physical methods to induce cell suspension as a positive control for anoikis and further examined the expression and cellular localization of Bit1 following NDV infection in tumor cells.The results indicated that both viral infection and cell suspension resulted in partial cell death,accompanied by the translocation of Bit1 from the mitochondria to the cytoplasm and a reduction in its protein levels.Notably,Bit1 expression was found not to significantly affect viral replication.These findings suggest that NDV infection promotes tumor cell death by activating Bit1 translocation,mirroring the effects observed during cell suspension-induced anoikis.In addition,in vivo experiments demonstrated that NDV effectively inhibits the metastasis and growth of melanoma in mice,and that overexpression of Bit1 in tumor cells accelerates this process.This study provides novel insights into NDV-induced tumor cell death and identifies potential targets for understanding the mechanisms of oncolytic virus action.展开更多
We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBO...We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBOV),which are intended for screening bats as well as other hosts and reservoirs of these three viruses.Our triplex NALFIA is a two-step assay format:the target nucleic acid in the sample is first amplified using tagged primers,and the tagged dsDNA amplicons are captured by antibodies immobilized on the NALFIA device,resulting in signal development from the binding of a streptavidin-colloidal gold conjugate to a biotin tag on the captured amplicons.Triplex amplification of the N gene of NiV,the UpE gene of MERS-CoV,and the Vp40 gene of REBOV was optimized,and three compatible combinations of hapten labels and antibodies were identified for end point detection.The lowest RNA copy numbers detected by the triplex NALFIA were 8.21e4 for the NiV N target,7.09e1 for the MERS-CoV UpE target,and 1.83e4 for the REBOV Vp40 target.Using simulated samples,the sensitivity and specificity for MERS-CoV and REBOV targets were estimated to be 100%,while the sensitivity and specificity for the NiV target were 91%and 93.3%,respectively.The compliance rate between triplex NALFIA and real-time RT‒PCR was 92%for the NiV N target and 100%for the MERS-CoV UpE and REBOV Vp40 targets.展开更多
TypeⅠinterferon(IFN)-mediated innate immune responses represent the first line of host defense against viral infection.However,the molecular mechanisms by which avian infuenza virus(AIV)inhibits typeⅠIFN production ...TypeⅠinterferon(IFN)-mediated innate immune responses represent the first line of host defense against viral infection.However,the molecular mechanisms by which avian infuenza virus(AIV)inhibits typeⅠIFN production in ducks are not well understood.Here,we frst found that the polymerase basic 2(PB2)protein of H5N1 subtype AIV inhibited the typeⅠIFN responses by targeting duck mitochondrial antiviral signaling protein(MAVS).We further demonstrated that H5N1-PB2 bound to theΔtransmembrane(ΔTM)domain of duck MAVS,and the polymerase basic 1(PB1)binding domain(PBD)and RNA binding nuclear import domain(RND)of H5N1-PB2 interacted with MAVS to inhibit typeⅠIFN expression in ducks.Collectively,our fndings contribute to understanding the molecular mechanism by which AIV proteins regulate the retinoic acid-inducible geneⅠ(RIG-Ⅰ)-like receptor(RLR)signaling pathway to evade host antiviral immune responses in ducks.展开更多
基金supported by grants from CAAST-ACLH(NAHEP/CAAST/2018-19)of the ICAR-World Bank-funded National Agricultural Higher Education Project(NAHEP).
文摘Background:Fruits of Indian gooseberry or Amla or Aonla(Phyllanthus emblica)are important ingredients in many Ayurvedic medicines,but little is known about its leaves.Methods:Different extracts and crude alkaloids of P.emblica leaves were evaluated for their antimicrobial activity against clinically important microbes using agar well diffusion assay.The antimicrobial activity of methanolic extract(ME)of P.emblica leaves was also compared with similarly prepared methanolic extracts from leaves of Yellow Kaner(Cascabela peruviana),Parijaat or Harsingar(Nyctanthes arbor-tristis),Custard apple(Annona squamosa),Garlic vine(Mansoa alliacea),Shami plant(Prosopis cineraria),Madar(Calotropis gigantea),and Bael(Aegle marmelos).Results:The ME of leaves of P.emblica was the most potent preparation against bacteria and yeast.Of the 338 strains of microbes belonging to 100 species(96 bacteria and four Candida species),the ME of P.emblica inhibited the growth of 300 strains.A total of 84.62%,96.39%,and 100%of 221 G−bacteria,111 G+bacteria,and 6 Candida species strains,respectively,were inhibited by ME of P.emblica leaves at≤36 mg/well.The aqueous extract of P.emblica leaves also inhibited a similar number of bacterial strains,but at higher concentrations,while the ether extract could inhibit only staphylococci.The alkaloid from P.emblica leaves and the ME from leaves of other plants had insignificant antimicrobial activity at similar≤36 mg/well concentration.Conclusion:The study concluded that the ME of P.emblica leaves may be a useful source of a potent,wide-spectrum antimicrobial substance(s).
基金supported by grants from CAAST-ACLH(NAHEP/CAAST/2018-19)of the ICAR-World Bank-funded National Agricultural Higher Education Project(NAHEP).
文摘Background:Achromobacter xylosoxidans(A.xylosoxidans)subspecies strains are known to be opportunistic environmental inhabitants.Among the two species,A.xylosoxidans ssp.xylosoxidans is more commonly reported cause of nosocomial infections colonizing the hospital environment and medical devices,while A.xylosoxidans ssp.denitrificans(AD)strains are widely distributed in the abiotic environment.The present retrospective observational study was aimed at understanding the occurrence of AD infections in the Bareilly region,and to look into the effective herbal and conventional antimicrobial resistance profile of the strains identified at the laboratory.Methods:The present retrospective study analysed Clinical Microbiology laboratory data of Indian veterinary research Institute.The data for the last 14 years(2011-2024)was retrieved,tabulated and analysed using MS Excel program to determine significance of occurrence,and variation in antimicrobial resistance of the strains isolated from different sources usingχ2 and odds ratio analysis.Results:The study revealed that AD was detectable as a potential pathogen not only from environmental samples but also from 51 clinical cases(either as pure culture or mixed infection),and also from healthy humans and animals.The pathogen was most commonly associated with deaths in animals and birds due to septicaemia and was isolated is single pathogen from blood samples.It was also detected as single pathogen from cases of abortions,metritis,and urinary tract infections.However,from cases of haemorrhagic enteritis,diarrhoea,mastitis,wound infections,pyoderma and abscesses,and middle ear infections AD was isolated in association of one or more potentially pathogenic bacteria.Of the 80 isolates in the study,68 had multiple drug resistance,and 21 produced metallo-β-lactamases responsible for resistance against most of theβ-lactam antibiotics,including cephalosporins and carbapenems.The most effective antibiotic was gentamicin,inhibiting 90.67%of the isolates,followed by tigecycline(85.00%),ciprofloxacin(80.77%),piperacillin tazobactam(80.65%);other antibiotics were effective against less than 80%of the isolates.Among the herbal antimicrobials,cinnamaldehyde,cinnamon oil,carvacrol,and ajowan oil inhibited 98.41%,85.07%,85.00%,and 83.75%of the isolates,respectively.Conclusion:The study concluded that in the Bareilly region in India,multiple-drug-resistance AD may be an emerging pathogen prevalent in environment and apparently healthy animals.More studies are warranted to understand the AD strains at molecular level to understand their zoonotic potential and circulation in the environment.
基金supported by the National KeyR&DProgramof China(2022YFF0710500)the National Natural Science Foundation of China(32172853 and 32373013)the Central Public-interest Scientific Institution Basal Research Fund,China(1610302022001).
文摘The Salmonella pathogenicity islands(SPIs) play crucial roles in the progression of Salmonella infection. In this study, we constructed an improved λ Red homologous recombination system to prepare single and triple deletion mutants of 3 prominent SPIs(SPI-1, 2, and 3), aiming at the impact of deletion on morphology, carbon source metabolism, adhesion and invasion capacity, in vivo colonization, and immune efficacy in chicks. Our examination revealed that the surface of the single deletion mutants(SM6ΔSPI1, ΔSPI2, and ΔSPI3) exhibited a more rugged texture and appeared to be enveloped in a layer of transparent colloid, whereas the morphology of the triple deletion mutant(SM6ΔSPI1&2&3) remained unaltered when compared to the parent strain. The carbon metabolic spectrum of the SPI mutants underwent profound alterations, with a notable and statistically significant modification observed in 30 out of 95 carbon sources, primarily carbohydrates(17 out of 30). Furthermore, the adhesion capacity of the 4 mutants to Caco-2 cells was significantly reduced when compared to that of the parent strain. Moreover,the invasion capacity of mutants SM6ΔSPI1 and SM6ΔSPI1&2&3 exhibited a substantial decrease, while it was enhanced to varying degrees for SM6ΔSPI3 and SM6ΔSPI2. Importantly, none of the 4 mutants induced any clinical symptoms in the chicks. However, they did transiently colonize the spleen and liver. Notably, the SM6ΔSPI1&2&3mutant was rapidly cleared from both the spleen and liver within 8 days post-infection and no notable pathological changes were observed in the organs. Additionally, when challenged, the mutants immunized groups displayed a significant increase in antibody levels and alterations in the CD3+CD4+ and CD3+CD8+ subpopulations, and the levels of IL-4 and IFN-γ cytokines in the SM6ΔSPI1&2&3 immunized chicken serum surpassed those of other groups.In summary, the successful construction of the 4 SPI mutants lays the groundwork for further exploration into the pathogenic(including metabolic) mechanisms of SPIs and the development of safe and effective live attenuated Salmonella vaccines or carriers.
基金supported by the National Key Research and Development Program of China(2021YFD1800402)the National Natural Science Foundation of China(32172856,31972654 and 32302881)+1 种基金the Natural Science Foundation of Shanghai,China(22ZR1476100 and 23ZR1476600)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(SHVRI-ASTIP-2014-8)。
文摘The inappropriate use of cephalosporins lead to the occurrence and global spread of bacteria resistant to these antimicrobials.In this study,we isolated four Escherichia albertii strains from broilers in eastern China.The antimicrobial susceptibility and genomic characterization of these E.albertii isolates were determined.Our results revealed that these four E.albertii isolates exhibited resistance to tetracyclines,chloramphenicol,β-lactams,aminoglycosides,polymyxin B,sulfonamides,quinolones,and other antimicrobials.Among them,EA04 isolate was multidrug resistant and harbored extended-spectrumβ-lactamases(ESBL)genes blaCTX-Mand blaTEM.Whole genome sequencing and core-genome multilocus sequence typing(cgMLST)based on all ST4638 E.albertii for EA04 inferred highly probable epidemiological links between selected human isolates.Additionally,the ESBL genes blaTEM-141and blaCTX-M-55were coexistent in an approximately 75 kb Inc FII plasmid pEA04.2 in EA04.Comparative analysis indicated that genes blaTEM-141and blaCTX-M-55were located in IS15-blaCTX-M-55-wbu C-blaTEM-141-IS26 region,which similar structures were identified in various bacteria.Furthermore,the plasmid pEA04.2 could be transferable to E.coli EC600 and lead to the resistance to third-generation cephalosporins.These results suggested that chicken potentially serve as a reservoir for multidrug resistant E.albertii,which increases the risk of horizontal transfer of antimicrobial resistance between humans,animals and environment.
基金supported by the National Key R&D Program of China(2022YFF0710500)the the National Natural Science Foundation,China(31802192,32172853 and 32373013)+2 种基金the Natural Science Foundation of Heilongjiang Province of China(C2018070)China Postdoctoral Science Foundation(2017M620076)the Central Public-interest Scientific Institution Basal Research Fund,China(1610302022001)。
文摘Salmonella enterica serovar Typhimurium,the causative agent of gastroenteritis,is one of the most successful intracellular pathogens.Although certain host factors for Salmonella infection have been unveiled,the factors mediating Salmonella entry,particularly the invasion process,remain obscure.Here,we have unearthed β2 integrin,a crucial member of the integrin family,as an important host factor facilitating Salmonella invasion.It is demonstrated that overexpression of β2 integrin promotes Salmonella invasion,while the knockdown of β2 integrin significantly diminishes the extent of invasion.Moreover,Salmonella exhibits specific binding affinity towards β2 integrin,and the block of β2 integrin on cell surface substantially reduces the infection of cells in vitro.The ectodomain soluble protein of β2 integrin neutralized Salmonella infection both in cells(in vitro)and in mice(in vivo).Additionally,Salmonella protein YrbD directly interacts with β2 integrin to facilitate its invasion.To our knowledge,this study showed for the first time that the protein YrbD mediates Salmonella adhesion and internalization into host cells by interacting with β2 integrin.These findings not only broaden our understanding of the mechanisms underlying Salmonella entry,but also identify a prospective target for therapeutic control.
基金supported by the National Natural Science Foundation of China(Grant no.32402927).
文摘As an emerging genotype,the G9 genotype rotaviruses(RVs)are widespread among humans and pigs,and have been reported in many countries and regions in the recent years.Moreover,porcine G9 strains could cross the interspecies barrier to infect human.To investigate the epidemic trends of porcine G9 strains as well as the cross-immunoreactivity among different isolates,an epidemiological investigation about porcine G9 genotype RVs(PoRVs)was performed during the period 2020-2023 in multiple provinces of China.A total of nine representative strains were identified.The phylogenetic analysis based on viral VP7 gene showed that these strains mainly clustered with lineages Ⅲ and Ⅵ,which revealed the predominant G9 PoRVs in China.Moreover,a new lineage,lineage Ⅶ,was identified,and strains of this lineage were found to be circulating in Guangdong and Taiwan.Except lineages Ⅰ and Ⅳ,some isolates from other lineages could co-circulate in pigs and humans.Three G9 strains,namely 923H,923E,and 923X,which belonged to the largest sub-lineage Ⅲ,were isolated.Then,the significant cross-reactivity was observed among strains of the same or different lineages.This study is the first to systematically investigate the genetic and immunogenetic characteristics of porcine G9 genotype rotavirus in China,as well as the potential cross-species transmission between pigs and humans,providing a valuable direction for the effective prevention of porcine rotavirus.
基金supported by grants from the National Key Research and Development Program of China(Grant No.2022YFD1800100)National Natural Science Foundation of China(Grant No.32272982)Natural Science Foundation of Shanghai(Grant No.23ZR1477100).
文摘Dear editor,As of 2023,the domestic cat population in China reached 65 million,surpassing dogs to become the most numerous companion animal in the country.Feline calicivirus(FCV)infection,one of the three most prevalent infectious diseases in cats,poses a severe threat to feline health.FCV,classified under the Caliciviridae family(genus Vesivirus).
基金supported by the grants from the open competition program of top ten critical priorities of Agricultural Science and Technology Innovation for the 14th Five-Year Plan of Guangdong Province(2024KJ14)the Fundamental Research Funds for the Central Universities(lzujbky-2022-ct02)+7 种基金the Project of National Center of Technology Innovation for Pigs(NCTIP-XD/C03)the Youth Innovation Program of the Chinese Academy of Agricultural Sciences(Y2025QC33)the Major Science and Technology Project of Gansu Province(22ZD6NA001 and 22ZD6NA012)the Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-CSLPDCP-2023002 and CAAS-ASTIP-2025-LVRI)the China Postdoctoral Science Foundation(2023M743830)the Earmarked Fund for CARS-35 and CARS-39-13he Fundamental Research Funds for Innovation Team of Gansu Province(23JRRA546,23JRRA548)the Basic Scientific Research Fund of LVRI(1610312021009).
文摘African swine fever(ASF)is an acute,hemorrhagic,and highly contagious disease in pigs caused by the African swine fever virus(ASFV).Our previous studies have demonstrated that deletion of the MGF360-9L gene weakens ASFV virulence in pigs,yet the underlying mechanism remains unclear.To investigate the mechanism of MGF360-9L regulating ASFV pathogenicity,the relationship between MGF360-9L and host proteins was identified by mass spectrometry.We found that host protein DEAD-box helicase 20(DDX20)interacted with and colocalized with MGF360-9L.Overexpression of DDX20 inhibited ASFV replication,whereas knockdown of DDX20 had the opposite effects.Moreover,DDX20 inhibited ASFV replication by promoting the activation of type I interferon signaling.Surprisingly,DDX20 was gradually degraded following ASFV infection.Mechanistically,MGF360-9L promoted the autophagic degradation of DDX20 by recruiting autophagy-related protein Ras-related protein Rab-1A(Rab1A).Silencing Rab1A suppressed ASFV replication,while overexpression of Rab1A exhibited the opposite effects.Furthermore,Rab1A,MGF360-9L and DDX20 could form a complex to facilitate the degradation of DDX20.Knockdown of Rab1A impaired MGF360-9L-mediated degradation of DDX20 during ASFV infection.In summary,our study demonstrates that MGF360-9L targets DDX20 for autophagy degradation to antagonize its antiviral function and facilitate ASFV replication.This finding broadens our understanding of the regulatory network between ASFV and its host,and provides new insights into the pathogenesis and immune evasion mechanisms of ASFV.
基金supported by grants received from CAAST-ACLH(NAHEP/CAAST/2018-19)of ICAR-World Bank-funded National Agricultural Higher Education Project(NAHEP).
文摘Background:Although many sugars are known antibacterial in higher concentrations(>50%W/V)and a few at low concentrations too(≤1 mg/mL),most of the studies are limited to only a few reference isolates,therefore no concrete conclusion can be drawn.Methods:This study evaluated the antimicrobial potential of 19 sugars against 8000 isolates of bacteria belonging to 46 genera and also against 30 reference strains of microbes of 14 different species.To determine susceptibility to sugars,a sugar-disc(1 mg)diffusion assay was performed on Mueller Hinton agar using the same method used for antimicrobial susceptibility of microbial strains.Results:In the study,of 3,336 isolates of Gram-positive bacteria 147(4.35%)were susceptible to one or more sugars but only 16(0.37%)of the 4644 isolates of Gram-negative bacteria were susceptible to one or other sugar.Gram-positive bacteria were significantly more often susceptible to one or more sugars than isolates of Gram-negative bacteria(OR 13.33,CI99,6.74–26.37).A total of 163 test-isolates(2.04%)but none of the reference strains were sugar susceptible.Most of the isolates susceptible to sugars(135 of 2,295)were members of the Bacillaceae(36/679)and Micrococcaceae(99/1,616)family.However,out of 5,705 isolates belonging to other bacterial families,only 28 isolates(0.49%)were susceptible to one or more sugars.The most effective to least effective sugars as antibacterial were mannose,inositol,mannitol,sucrose,raffinose,ribose,xylose,trehalose,dulcitol,maltose,lactose,inulin,salicin,melibiose,sorbitol,adonitol,arabinose,glucose,and esculin,inhibiting 69,58,23,14,11,10,10,7,7,7,6,6,5,5,5,5,2,1,and 1 of the test-isolates,respectively.Conclusion:The results are still intriguing in determining the utility of sugar susceptibility of different bacteria and are still beyond making any conclusion for their therapeutic utility.However,the study can be concluded that Gram-positive bacteria are generally more susceptible to lower concentrations of different sugars than Gram-negative bacteria,and various sugars have variable selectivity in their antibacterial effect on multiple types of bacteria.
基金supported by the grants from the National Key Research and Development Program of China(2022YFD1800604)the China Agriculture Research System(CARS-41)the Heilongjiang Touyan Innovation Team Program,China。
文摘Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase.Full-length cDNA of the LN16-A strain was constructed by assembling 5 cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme.Transfection of this plasmid,along with the supporting plasmids encoding the N,P,M2-1,and L proteins of LN16-A into BSR-T7/5 cells,resulted in the recovery of aMPV subtype B.To identify an effective insertion site,the enhanced green fluorescent protein(EGFP)gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As.The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites(between the leader and N genes or replacing the SH gene).To verify the availability of the site between G and L for foreign gene expression,the VP2 gene of very virulent infectious bursal disease virus(vvIBDV)was inserted into this site,and recombinant LN16-A(rLN16A-vvVP2)was successfully rescued.Single immunization of specificpathogen-free chickens with rLN16A-vvVP2 induced high levels of neutralizing antibodies and provided 100%protection against the virulent aMPV subtype B and vvIBDV.Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.
基金supported by grants from the National Key R&D Program of China(2022YFD1801000)the Natural Science Foundation of Shanghai,China(24ZR1479200)。
文摘Infectious bursal disease(IBD)is an acute,highly contagious disease that affects chicks(Müller et al.2003).IBD mainly damages the immune organs of chicks,especially the central immune organ,causing immune suppression in diseased chicks(Muller et al.2012).The pathogenic infectious bursal disease virus(IBDV)is a member of the Avira virus genus in the Birnaviridae family.(Harkness et al.1975;Dobos et al.1979;Müller et al.1979).IBDV is prevalent worldwide,causing serious economic losses to the global poultry industry.Currently,vaccination remains the most cost-effective way to prevent IBDV.
基金supported by funding from the National Natural Science Foundation of China(U23A20243 and 32272972 to QZ,32172820 to SX)the Major Science and Technology Project of Gansu Province(22ZD6NA001 to SX)+1 种基金the Youth Innovation Program(Y2023QC30)the Agricultural Science and Technology Innovation Program(CAAS-ASTIP-JBGS-20210102 to SX)of the Chinese Academy of Agricultural Sciences.
文摘Influenza A viruses(IAVs)are single-stranded negative-sense RNA viruses that continually challenge animal and human health.In IAV-infected cells,host RNA-binding proteins play key roles in the life cycle of IAV by directly binding to viral RNA.Here,we examined the role of the host RNA-binding protein nucleophosmin-1(NPM1)in IAV replication.We found that,as a nucleolar phosphoprotein,NPM1 directly binds to viral RNA(vRNA)and inhibits the replication of various subtypes of IAV.NPM1 binding to vRNA competitively reduces the assembly of the viral ribonucleoprotein complex and the viral polymerase activity,thereby reducing the generation of progeny viral RNA and virions.The RNA-binding activity of NPM1,with the key residues T199,T219,T234,and T237,is essential for its anti-influenza function.Taken together,our findings demonstrate that NPM1 acts as an RNA-binding protein and interacts with IAV vRNA to suppress viral replication.
基金supported by grants from the Heilongjiang Provincial Natural Science Foundation of China(LH2022 C109 to DQ L)the National Natural Science Foundation of China(32372985 to YZ L)+1 种基金the National Key Research and Development Program of China(2023YFD1802500 to YZ L)the Tianchi Talent Introduction Plan(IWA2023 to XJ W).
文摘In this study,we developed a highly sensitive enzyme-linked immunosorbent assay(ELISA)using newly produced monoclonal antibodies(mAbs)for detecting horse/donkey IL-1βin cell culture medium and serum samples.The mAbs were generated via the use of a KLH-conjugated peptide and purified equine IL-1βprotein as separate immunogens.Notably,the generated mAbs(3G8 and 5G3)demonstrated no cross-reactivity with other major inflammatory mediators,including IL-1α,IL-1Ra,TNF-α,and SAA.The IL-1βassay,which is based on the screened mAbs,exhibits a detection range of 200-10,000 pg/mL,meeting clinical detection requirements.The coefficients of variation for the repeatability and reproducibility of the assay were both less than 5%,indicating an acceptable level of variation.Subsequently,84 equine and 24 asinine serum samples were collected,and the IL-1βconcentration was measured with both our assay and a commercial kit in parallel.Our results revealed no significant difference between the in-house and commercial ELISA kits for the detection of IL-1βconcentrations in horse sera.Moreover,our ELISA method demonstrated superior sensitivity for IL-1βdetection in donkey samples compared to existing commercial assays.These findings suggest that the newly developed ELISA provides a reliable analytical method for detecting IL-1βin both equine and asinine samples.
基金This research was carried out within the State Assignment of the Ministry of Science and Higher Education of the Russian Federation(project FZGW-2024-0003).
文摘Objective:The objective of this study was to determine the level of methotrexate(MTX)toxicity in the intestines of mice and to evaluate the protective effect of probiotics composed of Streptococcus,Bifidobacterium,and Lactobacillus species on intestinal cells during MTX treatment.Methods:Mice were divided into three groups:control,MTX group(received MTX injections),and MTX+probiotics group(received MTX injections along with a diet containing probiotics).Morphological and histological changes,the level of mitochondrial DNA(mtDNA)damage,the level of lipid peroxidation products,and gene expression in the mice’s small intestine were assessed.Results:We demonstrated that intraperitoneal MTX injections significantly increased mtDNA damage in the liver(p<0.001),small intestine(p<0.001),and blood of mice(p<0.01).MTX elevated the quantity of lipid peroxidation products in the liver and small intestine,indicating its strong prooxidative properties.MTX induced structural changes in the mice’s intestines,characterized by leukocytic infiltration of tissues.Probiotic therapy in mice partially mitigated the morphological and histological changes in the small intestine induced by MTX,reduced oxidative stress,and promoted increased expression of quinone oxidoreductase 1(Nqo1),which participates in both cell protection against oxidative stress and drug/xenobiotic detoxification.Probiotics prevented the upregulation of the proinflammatory cytokine IL-1b in the small intestine and induced increased expression of genes associated with the Nuclear factor erythroid 2-related factor 2/Antioxidant response element(Nrf2/ARE)pathway,an important mechanism of cell protection.Conclusions:Probiotics can be considered an effective approach to reducing the toxicity of MTX during psoriasis or cancer treatment.
基金financially supported by the National Key Research and Development Program of China(No.2023YFD1800403 and 2023YFD1800404)。
文摘Monkeypox virus(MPXV),a member of the Orthopoxvirus genus,caused a large-scale global outbreak in 2022.Developing mouse models for MPXV infection is crucial for advancing research on vaccines and therapeutic interventions.To address this,we conducted a comparative study on the susceptibility of six mouse strains—severe combined immune-deficiency(SCID),nude,genetically diabetic(db/db)and obese(ob/ob),C57BL/6J,and BALB/c—to MPXV infection.Mouse strains were infected with MPXV via intranasal inoculation,and body weight changes and mortality were monitored post-infection.Additionally,the tissue distribution of MPXV and the pathological changes in the lung tissues of the infected mice were evaluated.The results demonstrated that SCID and nude mice exhibited significant weight loss following MPXV infection,with 100%mortality observed in SCID mice,while no mortality occurred in nude mice.In contrast,the other mouse strains showed no significant weight loss or mortality.Notably,the viral load in the lung tissues of SCID and nude mice was the highest among the tested strains.Furthermore,we investigated the impact of different inoculation routes—intranasal(I.N.),intraperitoneal(I.P.),and intravenous(I.V.)—on the pathogenicity of MPXV in mice.The results revealed that the intravenous route induced more pronounced pathogenic effects compared to the intranasal and intraperitoneal routes.In summary,this study provides valuable insights into the development of MPXV-infected mouse models,offering a foundation for further research on MPXV pathogenesis and therapeutic drug development.
基金supported by the earmarked fund for National Natural Science Foundation of China(32230105)the China Agriculture Research System(CARS-41)the National Major Agricultural Science and Technology Projects,China(NK2022100103)。
文摘Avian leukosis is an important tumorigenic disease caused by the avian leukosis virus(ALV)in poultry.ALVs belong to the retroviral family and are classified into 11 subgroups(ALV-A to ALV-K).Among them,ALV-J was first introduced into China in 1999,spreading widely and evolving from infecting meat-type chickens to layer chickens and Chinese local chickens.ALV-J typically induces myeloid leukosis in infected chickens,but also induces a high proportion of hemangiomas in infected layer chickens,posing a serious threat to poultry breeds in China.As a retrovirus,the genome of ALV-J has undergone significant mutations,which may be related to the expansion of the infection host range and increased pathogenicity of ALV-J.Over the last two decades,the introduction and spread of ALV-J in China have caused substantial losses to the poultry industry.Specialized detection assays have been developed to combat ALV-J infections in China.Additionally,ongoing research aims to employ gene-editing technology as a novel antiviral strategy to control the spread of ALV infections.This review highlights the importance of understanding the impact of ALV-J on the Chinese poultry industry and emphasizes the need for ongoing research and innovation to safeguard poultry health and promote sustainable poultry farming practices in China.
基金funded by the National Key Research and Development Program of China(2022YFD1800500,2021YFD1801401,2023YFD1802600)the Central Public-interest Scientific Institution Basal Research Fund,China(Y2022PT11)the Shanghai Sailing Program,China(23YF1457400).
文摘African swine fever(ASF),caused by African swine fever virus(ASFV),is a highly contagious swine disease that has spread globally.Effective control strategies are not yet available.In this study,we prepared K205R mRNA,which was then formulated using Lipid Nanoparticle(LNP).The resulting K205R mRNA-LNP showed a particle size of approximately 86.27 nm and an mRNA encapsulation efficiency of 96.24%.Efficient expression of the K205R protein was confirmed in both HEK293T and PK15 cells.We further evaluated the immunogenicity of K205R mRNA-LNP in mice and pigs.All immunized animals developed significantly higher levels of IgG antibodies against K205R compared to the control group in the first week after the second immunization,with antibody titers reaching up to 105.Challenge experiments showed that K205R mRNA delayed the time of death.Our results suggested the successful implementation of the mRNA platform in the preparation and application of ASFV mRNA.
基金supported by the National Key Research and Development Program of China(2022YFD1801500)the International Cooperation Project of National Natural Science Foundation of China(32220103012)the Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-CSLPDCP-202402).
文摘Anoikis is a specialized form of programmed cell death triggered by the detachment of cells from the extracellular matrix(ECM).Tumor cells that develop resistance to anoikis acquire the ability to detach,migrate,and colonize distant sites,ultimately leading to the formation of metastatic tumors.Bit1(Bcl-2 inhibitor of transcription 1),a key effector of anoikis,is released into the cytoplasm upon loss of cell attachment and activates a caspase-independent pathway of apoptosis.Newcastle disease virus(NDV),a pathogen that poses a significant threat to the poultry industry,has also emerged as a promising oncolytic virus capable of selectively targeting and killing tumor cells.However,whether NDV can induce the death of anoikis-resistant tumor cells by activating Bit1 remains unclear.In this study,we utilized physical methods to induce cell suspension as a positive control for anoikis and further examined the expression and cellular localization of Bit1 following NDV infection in tumor cells.The results indicated that both viral infection and cell suspension resulted in partial cell death,accompanied by the translocation of Bit1 from the mitochondria to the cytoplasm and a reduction in its protein levels.Notably,Bit1 expression was found not to significantly affect viral replication.These findings suggest that NDV infection promotes tumor cell death by activating Bit1 translocation,mirroring the effects observed during cell suspension-induced anoikis.In addition,in vivo experiments demonstrated that NDV effectively inhibits the metastasis and growth of melanoma in mice,and that overexpression of Bit1 in tumor cells accelerates this process.This study provides novel insights into NDV-induced tumor cell death and identifies potential targets for understanding the mechanisms of oncolytic virus action.
基金funded by the Department of Biotechnology,Ministry of Science and Technology,Government of India(DBT)under grant number ADMaC DBT-NER/LIVS/11/2012.
文摘We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBOV),which are intended for screening bats as well as other hosts and reservoirs of these three viruses.Our triplex NALFIA is a two-step assay format:the target nucleic acid in the sample is first amplified using tagged primers,and the tagged dsDNA amplicons are captured by antibodies immobilized on the NALFIA device,resulting in signal development from the binding of a streptavidin-colloidal gold conjugate to a biotin tag on the captured amplicons.Triplex amplification of the N gene of NiV,the UpE gene of MERS-CoV,and the Vp40 gene of REBOV was optimized,and three compatible combinations of hapten labels and antibodies were identified for end point detection.The lowest RNA copy numbers detected by the triplex NALFIA were 8.21e4 for the NiV N target,7.09e1 for the MERS-CoV UpE target,and 1.83e4 for the REBOV Vp40 target.Using simulated samples,the sensitivity and specificity for MERS-CoV and REBOV targets were estimated to be 100%,while the sensitivity and specificity for the NiV target were 91%and 93.3%,respectively.The compliance rate between triplex NALFIA and real-time RT‒PCR was 92%for the NiV N target and 100%for the MERS-CoV UpE and REBOV Vp40 targets.
基金supported by the grants from the National Natural Science Foundation of China(31872497 and 32072844)the National Key Research and Development Program of China(2021YFD1800200 and 2016YFD0500207)the Laboratory of Lingnan Modern Agriculture Project,China(NT2021007)。
文摘TypeⅠinterferon(IFN)-mediated innate immune responses represent the first line of host defense against viral infection.However,the molecular mechanisms by which avian infuenza virus(AIV)inhibits typeⅠIFN production in ducks are not well understood.Here,we frst found that the polymerase basic 2(PB2)protein of H5N1 subtype AIV inhibited the typeⅠIFN responses by targeting duck mitochondrial antiviral signaling protein(MAVS).We further demonstrated that H5N1-PB2 bound to theΔtransmembrane(ΔTM)domain of duck MAVS,and the polymerase basic 1(PB1)binding domain(PBD)and RNA binding nuclear import domain(RND)of H5N1-PB2 interacted with MAVS to inhibit typeⅠIFN expression in ducks.Collectively,our fndings contribute to understanding the molecular mechanism by which AIV proteins regulate the retinoic acid-inducible geneⅠ(RIG-Ⅰ)-like receptor(RLR)signaling pathway to evade host antiviral immune responses in ducks.
