Dear Editor,The highly pathogenic avian influenza viruses(HPAIVs)are important epizootic and zoonotic pathogens that cause significant economic losses to the poultry industry and pose a serious risk to veterinary and ...Dear Editor,The highly pathogenic avian influenza viruses(HPAIVs)are important epizootic and zoonotic pathogens that cause significant economic losses to the poultry industry and pose a serious risk to veterinary and public health.Wild birds have been recognized as the primary reservoirs for influenza A virus,and some species show little sign of clinical disease or even can be asymptomatic during long distance carriers of the virus(Lycett et al.,2019).Since it was first discovered in 1959,the H5Nx HPAIVs have spread globally and cause outbreaks in wild birds,poultry and sporadic human and other mammalian infections(Lycett et al.,2019).Due to the reassortant events of diverse strains facilitated by migratory waterfowl,the clade 2.3.4.4 of H5Nx viruses acquiring neuraminidase(NA)gene from other low pathogenicity avian influenza viruses(LPAIVs)emerged in 2014 and gradually became the dominant sub-clade(Lee et al.,2017).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
Hypertrophic cardiomyopathy(HCM)is an autosomal dominant inherited cardiomyopathy characterized by left ventricular hypertrophy.It is one of the chief causes of sudden cardiac death in younger people and athletes.Mole...Hypertrophic cardiomyopathy(HCM)is an autosomal dominant inherited cardiomyopathy characterized by left ventricular hypertrophy.It is one of the chief causes of sudden cardiac death in younger people and athletes.Molecular-genetic studies have confirmed that the vast majority of HCM is caused by mutations in genes encoding sarcomere proteins.HCM has a relatively wide phenotypic heterogeneity,varying from asymptomatic to sudden cardiac death,because of the many different mutations and pathogenic genes underlying it.Many studies have explored the clinical symptoms and prognosis of HCM,emphasizing the importance of genotype in evaluating patient prognosis and guiding the clinical management of HCM.To elaborate the main pathogenic genes and phenotypic prognosis in HCM to promote a better understanding of this genetic disease.Retrospective analysis of literature to evaluate the association between underlying gene mutations and clinical phenotypes in HCM patients.As sequencing technology advances,the pathogenic gene mutation spectrum and phenotypic characteristics of HCM are gradually becoming clearer.HCM is a widespread inherited disease with a highly variable clinical phenotype.The precise mechanisms linking known pathogenic gene mutations and the clinical course of this heterogeneous condition remain elusive.展开更多
Coffee wilt represents one of the most devastating diseases of Arabica coffee(Coffea arabica L.)plantations in the primary coffee-producing regions.In this study,coffee trees manifesting wilt symptoms accompanied by t...Coffee wilt represents one of the most devastating diseases of Arabica coffee(Coffea arabica L.)plantations in the primary coffee-producing regions.In this study,coffee trees manifesting wilt symptoms accompanied by the defoliation and drying of the whole tree were observed in the Jazan,El Baha,Najran,and Asir regions.The purpose of this investigation was to isolate and identify the Fusarium species recovered from symptomatic coffee trees.The developed fungi were initially characterized based on their morphological features followed by molecular phylogenetic multi-locus analysis of the combined sequences of ITS,TEF1-α,RPB2,and CaM.Twenty-five isolates were recovered from 28 samples.All fungal isolates were categorized morphologically under the genus Fusarium.Phylogenetic analysis positioned all the representative 15 isolates into one cluster grouping together with Neocosmospora falciformis(formerly F.falciforme)confirming their taxonomic position.Pathogenicity tests of the N.falciformis isolates were subsequently conducted on coffee seedlings,and the results revealed that all isolates induced wilt symptoms resembling those recorded in the field,and the incidence was 100%.The fungicide sensitivity test of seven investigated fungicides revealed that Maxim XL^(®) followed by Moncut^(®) exhibited the highest inhibitory effect against N.falciformis KSA 24-14,reaching 93.33%and 91.67%,respectively.To our knowledge,N.falciformis is a new causal pathogen of coffee wilt in Saudi Arabia.Remarkably,these results offer important insights for devising effective approaches to monitor and control such diseases.展开更多
Grape white rot caused by Coniella vitis is a global concern in the grape industry.pH regulation is essential for cell growth,reproductive processes and pathogenicity in phytopathogenic fungi.In this study,we observed...Grape white rot caused by Coniella vitis is a global concern in the grape industry.pH regulation is essential for cell growth,reproductive processes and pathogenicity in phytopathogenic fungi.In this study,we observed that the growth rate,spore production and virulence of C.vitis significantly declined in alkaline pH,as well as the suppressive effect on secretion of hydrolytic enzymes.Transcriptomic and metabolomic analyses were used to investigate the responses of C.vitis to acidic(pH 5),neutral(pH 7)and alkaline environments(pH 9).We identified 728,1,780 and 3,386 differentially expressed genes(DEGs)at pH 5,pH 7 and pH 9,when compared with the host pH(pH 3),and 2,122 differently expressed metabolites(DEMs)in negative and positive ion mode.Most DEGs were involved in carbohydrate metabolic process,transmembrane transport,tricarboxylic acid cycle,peptide metabolic process,amide biosynthetic process,and organic acid metabolic process.In addition,metabolomic analysis revealed ABC transporters,indole alkaloid biosynthesis,diterpenoid biosynthesis,and carotenoid biosynthesis pathways in response to the pH change.Furthermore,we found that the aspartate synthesis metabolic route associated with the TCA cycle is a key limiting factor for the growth and development of C.vitis in alkaline environments,and aspartate supplementation enables C.vitis to grow in alkaline environments.Plant cell wall-degrading enzymes(PCWDEs)could contribute to the pathogenicity,when C.vitis infected at pH 3.Importantly,aflatrem biosynthesis in acidic environment might contribute to the virulence of C.vitis and has a risk of causing human health problems due to its acute neurotoxic effects.展开更多
Magnaporthe oryzae is the causal agent of rice blast. Glycosylation plays key roles in vegetative growth,development, and infection of M. oryzae. However, several glycosylation-related genes have not been characterize...Magnaporthe oryzae is the causal agent of rice blast. Glycosylation plays key roles in vegetative growth,development, and infection of M. oryzae. However, several glycosylation-related genes have not been characterized.In this study, we identified a Glyco_transf_22 domain-containing protein, MoAlg9, and found that MoAlg9 islocalized to the endoplasmic reticulum(ER). Deletion of MoALG9 significantly affected conidial production, normalappressorium formation, responses to stressors, and pathogenicity of M. oryzae. We also found that the ΔMoalg9mutant was defective in glycogen utilization, appressorial penetration, and invasive growth in host cells. Moreover,we further demonstrated that MoALG9 regulates the transcription of several target genes involved in conidiation,appressorium formation, and cell wall integrity. In addition, we found that the Glyco_transf_22 domain is essentialfor normal MoAlg9 function and localization. We also provide evidence that MoAlg9 is involved in N-glycosylationpathway in M. oryzae. Taken together, these results show that MoAlg9 is important for conidiation, appressoriumformation, maintenance of cell wall integrity, and the pathogenesis of M. oryzae.展开更多
Plant pathogens secrete various cell wall-degrading enzymes that compromise host cell wall integrity and facilitate pathogen invasion.This study identified VdGH7a,a glycoside hydrolase family 7(GH7)cellobiohydrolase f...Plant pathogens secrete various cell wall-degrading enzymes that compromise host cell wall integrity and facilitate pathogen invasion.This study identified VdGH7a,a glycoside hydrolase family 7(GH7)cellobiohydrolase from Verticillium dahliae,which demonstrated hydrolytic activity against 1,4-β-glucan.Notably,VdGH7a induced cell death in Nicotiana benthamiana when signal peptides were present,though this effect was inhibited by the carbohydrate-binding type-1(CBM1)protein domain.The deletion of VdGH7a substantially reduced V.dahliae pathogenicity in cotton plants,as demonstrated by the mutants’inability to penetrate cellophane membrane.These knockout mutants also exhibited reduced carbon source utilization capacity and increased sensitivity to osmotic and cell wall stresses.Through yeast two-hybrid screening,bi-molecular fluorescence complementation(BiFC),and luciferase complementation imaging(LCI),we identified that VdGH7a interacts with an osmotin-like protein(GhOLP1)in cotton.Virus-induced gene silencing of GhOLP1 resulted in decreased salicylic acid(SA)content and reduced resistance to V.dahliae in cotton,while heterologous overexpression of GhOLP1 in Arabidopsis enhanced both resistance and SA signaling pathway gene expression.These results reveal a virulence mechanism wherein the secreted protein VdGH7a from V.dahliae interacts with GhOLP1 to activate host immunity and contribute significantly to plant resistance against V.dahliae.展开更多
Background:New variants of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continue to drive global epidemics and pose significant health risks.The pathogenicity of these variants evolves under immune press...Background:New variants of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continue to drive global epidemics and pose significant health risks.The pathogenicity of these variants evolves under immune pressure and host factors.Understanding these changes is crucial for epidemic control and variant research.Methods:Human angiotensin-converting enzyme 2(hACE2)transgenic mice were in-tranasally challenged with the original strain WH-09 and the variants Delta,Beta,and Omicron BA.1,while BALB/c mice were challenged with Omicron subvariants BA.5,BF.7,and XBB.1.To compare the pathogenicity differences among variants,we con-ducted a comprehensive analysis that included clinical symptom observation,meas-urement of viral loads in the trachea and lungs,evaluation of pulmonary pathology,analysis of immune cell infiltration,and quantification of cytokine levels.Results:In hACE2 mice,the Beta variant caused significant weight loss,severe lung inflammation,increased inflammatory and chemotactic factor secretion,greater mac-rophage and neutrophil infiltration in the lungs,and higher viral loads with prolonged shedding duration.In contrast,BA.1 showed a significant reduction in pathogenicity.The BA.5,BF.7,and XBB.1 variants were less pathogenic than the WH-09,Beta,and Delta variants when infected in BALB/c mice.This was evidenced by reduced weight loss,diminished pulmonary pathology,decreased secretion of inflammatory factors and chemokines,reduced macrophage and neutrophil infiltration,as well as lower viral loads in both the trachea and lungs.Conclusion:In hACE2 mice,the Omicron variant demonstrated the lowest pathogenic-ity,while the Beta variant exhibited the highest.Pathogenicity of the Delta variant was comparable to the original WH-09 strain.Among BALB/c mice,Omicron subvari-ants BA.5,BF.7,and XBB.1 showed no statistically significant differences in virulence.展开更多
In Candida species,the endoplasmic reticulum(ER)stress response—regulated by the unfolded protein response(UPR)—serves as a critical adaptive mechanism affecting both pathogenicity and antifungal resistance.This rev...In Candida species,the endoplasmic reticulum(ER)stress response—regulated by the unfolded protein response(UPR)—serves as a critical adaptive mechanism affecting both pathogenicity and antifungal resistance.This review aims to synthesize current knowledge on ER stress pathways in Candida glabrata and Candida albicans,highlighting their species-specific adaptations and therapeutic implications.We systematically analyzed peer-reviewed literature on ER stress mechanisms in Candida,focusing on comparative studies of UPR signaling.Emphasis was placed on C.glabrata’s inositol-requiring enzyme 1(IRE1)-dependent Regulated IRE1-Dependent Decay(RIDD)pathway and C.albicans’IRE1/HAC1 and calcium-mediated pathways.Connections to virulence and drug resistance were evaluated through genetic,transcriptomic,and phenotypic evidence.Candida species employ divergent UPR strategies:C.glabrata mitigates ER stress primarily via RIDD-mediated mRNA decay to reduce protein load,while C.albicans enhances folding capacity through HAC1 splicing and calcium homeostasis.These adaptations promote survival in hostile host environments(e.g.,oxidative stress,immune attacks)and are linked to resistance against azoles and echinocandins.Pharmacological disruption of UPR components(e.g.,IRE1 inhibitors)sensitizes Candida to antifungals in experimental models.ER stress response pathways are promising targets for antifungal drug development.Understanding species-specific UPR mechanisms in Candida could guide novel therapies to overcome resistance and improve treatment outcomes.展开更多
The H4 subtype of avian influenza virus(AIV)is prevalent worldwide,but only receives little attention due to its low pathogenicity in poultry.Consequently,it remains largely unclear whether H4 AIVs pose a potential th...The H4 subtype of avian influenza virus(AIV)is prevalent worldwide,but only receives little attention due to its low pathogenicity in poultry.Consequently,it remains largely unclear whether H4 AIVs pose a potential threat to the poultry industry and public health.During the period from 2011 to 2022,we conducted an active surveillance programm.A total of 154,762 swab samples were collected across various provinces,and 427 H4 viruses were detected,resulting in a positivity rate of 0.28%.All H4 viruses were isolated from poultry,primarily from ducks in live poultry markets.We further investigated the genetic evolutionary characteristics and pathogenicity of 20 H4Nx viruses isolated in our program.Phylogenetic analysis revealed that the 20 H4Nx viruses belonged to the Eurasian lineage and exhibited significant genetic diversity,with 19 distinct genotypes identified.Molecular characterization indicated that these viruses were low-pathogenicity AIVs with limited binding affinity to human receptors,yet they contained mutations associated with enhanced viral replication and pathogenicity in mammals.Pathogenicity tests conducted in ducks demonstrated that H4 viruses were weakly pathogenic,exhibiting limited replication and transmission capabilities.However,some viruses were able to replicate effectively in mice and induce weight loss.For instance,DK/AH/AG61/11(H4N6)can replicate efficiently in MDCK cells,indicating a potential threat to mammals.These findings underscore the importance of ongoing surveillance of H4 AIVs to better understand their evolution and transmission dynamics and to prevent potential public health risks.展开更多
Avian pathogenic Escherichia coli(APEC)belonging to extraintestinal pathogenic E.coli(ExPEC)can cause severe infections in extraintestinal tissues in birds and humans,such as the lungs and blood.MprA(microcin producti...Avian pathogenic Escherichia coli(APEC)belonging to extraintestinal pathogenic E.coli(ExPEC)can cause severe infections in extraintestinal tissues in birds and humans,such as the lungs and blood.MprA(microcin production regulation,locus A,herein renamed AbsR,a blood survival regulator),a member of the MarR(multiple antibiotic resistance regulator)transcriptional regulator family,governs the expression of capsule biosynthetic genes in human ExPEC and represents a promising druggable target for antimicrobials.However,a deep understanding of the AbsR regulatory mechanism as well as its regulon is lacking.In this study,we present a systems-level analysis of the APEC AbsR regulon using ChIP-Seq(chromatin immunoprecipitation sequencing)and RNA-Seq(RNA sequencing)methods.We found that AbsR directly regulates 99 genes and indirectly regulates 667 genes.Furthermore,we showed that:1)AbsR contributes to antiphagocytotic effects by macrophages and virulence in a mouse model for systemic infection by directly activating the capsular gene cluster;2)AbsR positively impacts biofilm formation via direct regulation of the T2SS(type II secretion system)but plays a marginal role in virulence;and 3)AbsR directly upregulates the acid tolerance signaling system EvgAS to withstand acid stress but is dispensable in ExPEC virulence.Finally,our data indicate that the role of AbsR in virulence gene regulation is relatively conserved in ExPEC strains.Altogether,this study provides a comprehensive analysis of the AbsR regulon and regulatory mechanism,and our data suggest that AbsR likely influences virulence primarily through the control of capsule production.Interestingly,we found that AbsR severely represses the expression of the type I-F CRISPR(clustered regularly interspaced short palindromic repeats)-Cas(CRISPR associated)systems,which could have implications in CRISPR biology and application.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
Pathogenic microorganisms produce numerous metabolites,including volatile organic compounds(VOCs).Monitoring these metabolites in biological matrices(e.g.,urine,blood,or breath)can reveal the presence of specific micr...Pathogenic microorganisms produce numerous metabolites,including volatile organic compounds(VOCs).Monitoring these metabolites in biological matrices(e.g.,urine,blood,or breath)can reveal the presence of specific microorganisms,enabling the early diagnosis of infections and the timely implementation of tar-geted therapy.However,complex matrices only contain trace levels of VOCs,and their constituent com-ponents can hinder determination of these compounds.Therefore,modern analytical techniques enabling the non-invasive identification and precise quantification of microbial VOCs are needed.In this paper,we discuss bacterial VOC analysis under in vitro conditions,in animal models and disease diagnosis in humans,including techniques for offline and online analysis in clinical settings.We also consider the advantages and limitations of novel microextraction techniques used to prepare biological samples for VOC analysis,in addition to reviewing current clinical studies on bacterial volatilomes that address inter-species in-teractions,the kinetics of VOC metabolism,and species-and drug-resistance specificity.展开更多
The necrotrophic fungus, Sclerotinia sclerotiorum, employs an array of cell wall-degrading enzymes(CWDEs), including cellulase, to dismantle host cell walls. However, the molecular mechanisms through which S. scleroti...The necrotrophic fungus, Sclerotinia sclerotiorum, employs an array of cell wall-degrading enzymes(CWDEs), including cellulase, to dismantle host cell walls. However, the molecular mechanisms through which S. sclerotiorum degrades cellulose remain elusive. Here, we unveil a novel secretory cellobiohydrolase, SsdchA, characterized by a signal peptide and a Glyco_hydro_7(GH7) domain. SsdchA exhibits a robust expression of during early infection stages. Interestingly, colony morphology and growth rates remain unaffected across the wild-type, SsdchA deletion strains and SsdchA overexpression strains on potato dextrose agar(PDA) medium. Nevertheless, the pathogenicity and cellobiohydrolase activity decreased in the SsdchA deletion strains, but enhanced in the SsdchA overexpression strains. Moreover,the heterologous expression of SsdchA in Arabidopsis thaliana leads to reduced cellulose content and heightened susceptibility to S. sclerotiorum. Collectively, our data underscore the pivotal role of the novel cellobiohydrolase SsdchA in the pathogenicity of S. sclerotiorum.展开更多
Litchi downy blight,caused by the plant pathogenic oomycete Peronophythora litchii,is one of the most devastating diseases on litchi and resulted in huge economic losses.Autophagy plays an essential role in the develo...Litchi downy blight,caused by the plant pathogenic oomycete Peronophythora litchii,is one of the most devastating diseases on litchi and resulted in huge economic losses.Autophagy plays an essential role in the development and pathogenicity of the filamentous fungi.However,the function of autophagy in oomycetes remain elusive.Here,an autophagy-related protein Atg3 homolog PlAtg3 was identified and characterized in P.litchii.The absence of PlATG3 through the CRISPR/Cas9 gene replacement strategy compromised vegetative growth and sexual/asexual development.Cytological analyses revealed that the deletion of PlATG3 impaired autophagosome formation with monodansylcadaverine(MDC)staining and significantly disrupted zoospore release due to defects of sporangial cleavage with FM4-64 staining.Atg8 is considered to be an autophagy marker protein in various species.Western blot analysis indicated that PlAtg3 is involved in degradation of PlAtg8-PE.Interestingly,PlAtg3 was unable to interact with PlAtg8 in yeast two hybrid(Y2H)assays,possibly due to the absence of the Atg8 family interacting motif(AIM)in PlAtg3.Furthermore,pathogenicity assays revealed that the deletion of PlATG3 considerably reduced the virulence of P.litchii.Taken together,our data reveal that PlAtg3 plays an important role in radial growth,asexual/sexual development,sporangial cleavage and zoospore release,autophagosome formation,and pathogenicity in P.litchii.This study contributes to a better understanding of the pathogenicity mechanisms of P.litchii and provides insights for the development of more effective strategies to control oomycete diseases.展开更多
The stamping-out strategy has been used to control highly pathogenic avian influenza viruses in many countries,driven by the belief that vaccination would not be successful against such viruses and fears that avian in...The stamping-out strategy has been used to control highly pathogenic avian influenza viruses in many countries,driven by the belief that vaccination would not be successful against such viruses and fears that avian influenza virus in vaccinated birds would evolve more rapidly and pose a greater risk to humans.In this review,we summarize the successes in controlling highly pathogenic avian influenza in China and make suggestions regarding the requirements for vaccine selection and effectiveness.In addition,we present evidence that vaccination of poultry not only eliminates human infection with avian influenza virus,but also significantly reduces and abolishes some harmful characteristics of avian influenza virus.展开更多
AIM:To investigate the types of bacteria in patients with eye infections in Suzhou and their drug resistance to commonly used antibacterial drugs.METHODS:The clinical data of 155 patients were retrospectively collecte...AIM:To investigate the types of bacteria in patients with eye infections in Suzhou and their drug resistance to commonly used antibacterial drugs.METHODS:The clinical data of 155 patients were retrospectively collected in this study,and the pathogenic bacteria species and drug resistance of each pathogenic bacteria were analyzed.RESULTS:Among the 155 patients(age from 12 to 87 years old,with an average age of 57,99 males and 56 females)with eye infections(160 eyes:74 in the left eye,76 in the right eye and 5 in both eyes,all of which were exogenous),71(45.81%)strains were gram-positive bacteria,23(14.84%)strains were gram-negative bacteria and 61(39.35%)strains were fungi.Gram-positive bacteria were highly resistant to penicillin and erythromycin(78.87%and 46.48%respectively),but least resistant to vancomycin at 0.Gram-negative bacteria were highly resistant to cefoxitin and compound sulfamethoxazole(100%and 95.65%respectively),but least resistant to meropenem at 0.Comparison of the resistance of gram-positive and gram-negative bacteria to some drugs revealed statistically significant differences(P<0.05)in the resistance of both to cefoxitin,cotrimoxazole,levofloxacin,cefuroxime,ceftriaxone and ceftazidime,and both had higher rates of resistance to gram-negative bacteria than to gram-positive bacteria.The distribution of bacterial infection strains showed that Staphylococcus epidermidis was the most common strain in the conjunctiva,cornea,aqueous humor or vitreous body and other eye parts.Besides,Fusarium and Pseudomonas aeruginosa were also among the most common strains of conjunctival and corneal infections.CONCLUSION:Gram-positive bacteria are the dominant bacteria in eye infections,followed by gram-negative bacteria and fungi.Considering the resistance of gramnegative bacteria to multiple drugs,monitoring of bacteria should be strengthened in eye bacterial infections for effective prevention and control to reduce complications caused by eye infections.展开更多
Bordetella bronchiseptica(Bb)is recognized as a leading cause of respiratory diseases in dogs and cats.However,epidemiological data on Bb in dogs and cats in China are still limited,and there is no commercially availa...Bordetella bronchiseptica(Bb)is recognized as a leading cause of respiratory diseases in dogs and cats.However,epidemiological data on Bb in dogs and cats in China are still limited,and there is no commercially available vaccine.Live vaccines containing Bb that are widely used abroad are generally efective but can establish latency and potentially reactivate to cause illness in some immunodefcient vaccinated recipients,raising safety concerns.In this study,34 canine-derived and two feline-derived Bb strains were isolated from 1809 canine and 113 feline nasopharyngeal swab samples collected from eight provinces in China from 2021 to 2023.The PCR results showed that the percentage of positive Bb was 22.94%(441/1922),and more than 90%of the Bb isolates had four virulence factor-encoding genes(VFGs),namely,fhaB,prn,betA and dnt.All the isolated strains displayed a multidrug-resistant phenotype.The virulence of 10 Bb strains isolated from dogs with respiratory symptoms was tested in mice,and we found that eight isolates were highly virulent.Furthermore,the eight Bb isolates with high virulence were inactivated and intramuscularly injected into mice,and three Bb strains(WH1218,WH1203 and WH1224)with the best protective efcacy were selected.Dogs immunized with these three strains exhibited strong protection against challenge with the Bb feld strain WH1218.Ultimately,the WH1218 strain with the greatest protection in dogs was selected as the vaccine candidate.Dogs and cats that received a vaccine containing 109 CFU of the inactivated WH1218 strain showed complete protection against challenge with the Bb feld strain WH1218.This study revealed that Bb is an important pathogen that causes respiratory diseases in domestic dogs and cats in China,and all the isolates exhibited multidrug resistance.The present work contributes to the current understanding of the prevalence,antimicrobial resistance,and virulence genes of Bb in domestic dogs and cats.Additionally,our results suggest that the WH1218 strain is a promising candidate safe and efcacious inactivated Bb vaccine.展开更多
Liver transplantation(LT)is the standard therapy for individuals afflicted with end-stage liver disease.Despite notable advancements in LT technology,the incidence of early allograft dysfunction(EAD)remains a critical...Liver transplantation(LT)is the standard therapy for individuals afflicted with end-stage liver disease.Despite notable advancements in LT technology,the incidence of early allograft dysfunction(EAD)remains a critical concern,exacerbating the current organ shortage and detrimentally affecting the prognosis of recipients.Unfortunately,the perplexing hepatic heterogeneity has impeded characterization of the cellular traits and molecular events that contribute to EAD.Herein,we constructed a pioneering single-cell transcriptomic landscape of human transplanted livers derived from non-EAD and EAD patients,with 12 liver samples collected from 7 donors during the cold perfusion and portal reperfusion stages.Comparison of the 75231 cells of non-EAD and EAD patients revealed an EAD-associated immune niche comprising mucosal-associated invariant T cells,granzyme B^(+)(GZMB^(+))granzyme K^(+)(GZMK^(+))natural killer cells,and S100 calcium binding protein A12^(+)(S100A12^(+))neutrophils.Moreover,we verified this immune niche and its association with EAD occurrence in two independent cohorts.Our findings elucidate the cellular characteristics of transplanted livers and the EAD-associated pathogenic immune niche at the single-cell level,thus,offering valuable insights into EAD onset.展开更多
AIM:To describe the clinical,electrophysiological,and genetic features of an unusual case with an RDH12 homozygous pathogenic variant and reviewed the characteristics of the patients reported with the same variant.MET...AIM:To describe the clinical,electrophysiological,and genetic features of an unusual case with an RDH12 homozygous pathogenic variant and reviewed the characteristics of the patients reported with the same variant.METHODS:The patient underwent a complete ophthalmologic examination including best-corrected visual acuity,anterior segment and dilated fundus,visual field,spectral-domain optical coherence tomography(OCT)and electroretinogram(ERG).The retinal disease panel genes were sequenced through chip capture high-throughput sequencing and Sanger sequencing was used to confirm the result.Then we reviewed the characteristics of the patients reported with the same variant.RESULTS:A 30-year male presented with severe early retinal degeneration who complained night blindness,decreased visual acuity,vitreous floaters and amaurosis fugax.The best corrected vision was 0.04 OD and 0.12 OS,respectively.The fundus photo and OCT showed bilateral macular atrophy but larger areas of macular atrophy in the left eye.Autofluorescence shows bilateral symmetrical hypo-autofluorescence.ERG revealed that the amplitudes of a-and b-wave were severely decreased.Multifocal ERG showed decreased amplitudes in the local macular area.A homozygous missense variant c.146C>T(chr14:68191267)was found.The clinical characteristics of a total of 13 patients reported with the same pathologic variant varied.CONCLUSION:An unusual patient with a homozygous pathogenic variant in the c.146C>T of RDH12 which causes late-onset and asymmetric retinal degeneration are reported.The clinical manifestations of the patient with multimodal retinal imaging and functional examinations have enriched our understanding of this disease.展开更多
基金supported by Zhejiang Province Science and Technology Cooperation Project of“Three Rural and Nine Parties”(grant number 2023SNJF059).
文摘Dear Editor,The highly pathogenic avian influenza viruses(HPAIVs)are important epizootic and zoonotic pathogens that cause significant economic losses to the poultry industry and pose a serious risk to veterinary and public health.Wild birds have been recognized as the primary reservoirs for influenza A virus,and some species show little sign of clinical disease or even can be asymptomatic during long distance carriers of the virus(Lycett et al.,2019).Since it was first discovered in 1959,the H5Nx HPAIVs have spread globally and cause outbreaks in wild birds,poultry and sporadic human and other mammalian infections(Lycett et al.,2019).Due to the reassortant events of diverse strains facilitated by migratory waterfowl,the clade 2.3.4.4 of H5Nx viruses acquiring neuraminidase(NA)gene from other low pathogenicity avian influenza viruses(LPAIVs)emerged in 2014 and gradually became the dominant sub-clade(Lee et al.,2017).
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 National Natural Science Foundation of China,No.81770379 and 81470521.
文摘Hypertrophic cardiomyopathy(HCM)is an autosomal dominant inherited cardiomyopathy characterized by left ventricular hypertrophy.It is one of the chief causes of sudden cardiac death in younger people and athletes.Molecular-genetic studies have confirmed that the vast majority of HCM is caused by mutations in genes encoding sarcomere proteins.HCM has a relatively wide phenotypic heterogeneity,varying from asymptomatic to sudden cardiac death,because of the many different mutations and pathogenic genes underlying it.Many studies have explored the clinical symptoms and prognosis of HCM,emphasizing the importance of genotype in evaluating patient prognosis and guiding the clinical management of HCM.To elaborate the main pathogenic genes and phenotypic prognosis in HCM to promote a better understanding of this genetic disease.Retrospective analysis of literature to evaluate the association between underlying gene mutations and clinical phenotypes in HCM patients.As sequencing technology advances,the pathogenic gene mutation spectrum and phenotypic characteristics of HCM are gradually becoming clearer.HCM is a widespread inherited disease with a highly variable clinical phenotype.The precise mechanisms linking known pathogenic gene mutations and the clinical course of this heterogeneous condition remain elusive.
基金funded by the Deanship of Scientific Research,Vice Presidency for Graduate Studies and Scientific Research,King Faisal University,Saudi Arabia,for supporting this work for work through grant number KFU242134.
文摘Coffee wilt represents one of the most devastating diseases of Arabica coffee(Coffea arabica L.)plantations in the primary coffee-producing regions.In this study,coffee trees manifesting wilt symptoms accompanied by the defoliation and drying of the whole tree were observed in the Jazan,El Baha,Najran,and Asir regions.The purpose of this investigation was to isolate and identify the Fusarium species recovered from symptomatic coffee trees.The developed fungi were initially characterized based on their morphological features followed by molecular phylogenetic multi-locus analysis of the combined sequences of ITS,TEF1-α,RPB2,and CaM.Twenty-five isolates were recovered from 28 samples.All fungal isolates were categorized morphologically under the genus Fusarium.Phylogenetic analysis positioned all the representative 15 isolates into one cluster grouping together with Neocosmospora falciformis(formerly F.falciforme)confirming their taxonomic position.Pathogenicity tests of the N.falciformis isolates were subsequently conducted on coffee seedlings,and the results revealed that all isolates induced wilt symptoms resembling those recorded in the field,and the incidence was 100%.The fungicide sensitivity test of seven investigated fungicides revealed that Maxim XL^(®) followed by Moncut^(®) exhibited the highest inhibitory effect against N.falciformis KSA 24-14,reaching 93.33%and 91.67%,respectively.To our knowledge,N.falciformis is a new causal pathogen of coffee wilt in Saudi Arabia.Remarkably,these results offer important insights for devising effective approaches to monitor and control such diseases.
基金supported by the Shandong Provincial Natural Science Foundation,China(ZR2021QC131)the Shandong Province Key Research and Development Plan,China(2022TZXD001102)+1 种基金the Shandong Province Demonstration Project for Model Construction in Rural Revitalization Service,China(2022DXAL0226)the Innovation Project of Shandong Academy of Agricultural Sciences,China(CXGC2023F15,CXGC2023A41,and CXGC2023A47)。
文摘Grape white rot caused by Coniella vitis is a global concern in the grape industry.pH regulation is essential for cell growth,reproductive processes and pathogenicity in phytopathogenic fungi.In this study,we observed that the growth rate,spore production and virulence of C.vitis significantly declined in alkaline pH,as well as the suppressive effect on secretion of hydrolytic enzymes.Transcriptomic and metabolomic analyses were used to investigate the responses of C.vitis to acidic(pH 5),neutral(pH 7)and alkaline environments(pH 9).We identified 728,1,780 and 3,386 differentially expressed genes(DEGs)at pH 5,pH 7 and pH 9,when compared with the host pH(pH 3),and 2,122 differently expressed metabolites(DEMs)in negative and positive ion mode.Most DEGs were involved in carbohydrate metabolic process,transmembrane transport,tricarboxylic acid cycle,peptide metabolic process,amide biosynthetic process,and organic acid metabolic process.In addition,metabolomic analysis revealed ABC transporters,indole alkaloid biosynthesis,diterpenoid biosynthesis,and carotenoid biosynthesis pathways in response to the pH change.Furthermore,we found that the aspartate synthesis metabolic route associated with the TCA cycle is a key limiting factor for the growth and development of C.vitis in alkaline environments,and aspartate supplementation enables C.vitis to grow in alkaline environments.Plant cell wall-degrading enzymes(PCWDEs)could contribute to the pathogenicity,when C.vitis infected at pH 3.Importantly,aflatrem biosynthesis in acidic environment might contribute to the virulence of C.vitis and has a risk of causing human health problems due to its acute neurotoxic effects.
基金supported by the National Natural Science Foundation of China (32202253)the Natural Science Foundation of Anhui Higher Education Institutions, China (KJ2020A0102)the Talent Research Project of Anhui Agricultural University, China (rc342001)。
文摘Magnaporthe oryzae is the causal agent of rice blast. Glycosylation plays key roles in vegetative growth,development, and infection of M. oryzae. However, several glycosylation-related genes have not been characterized.In this study, we identified a Glyco_transf_22 domain-containing protein, MoAlg9, and found that MoAlg9 islocalized to the endoplasmic reticulum(ER). Deletion of MoALG9 significantly affected conidial production, normalappressorium formation, responses to stressors, and pathogenicity of M. oryzae. We also found that the ΔMoalg9mutant was defective in glycogen utilization, appressorial penetration, and invasive growth in host cells. Moreover,we further demonstrated that MoALG9 regulates the transcription of several target genes involved in conidiation,appressorium formation, and cell wall integrity. In addition, we found that the Glyco_transf_22 domain is essentialfor normal MoAlg9 function and localization. We also provide evidence that MoAlg9 is involved in N-glycosylationpathway in M. oryzae. Taken together, these results show that MoAlg9 is important for conidiation, appressoriumformation, maintenance of cell wall integrity, and the pathogenesis of M. oryzae.
基金supported by the Project of Sanya Yazhou Bay Science and Technology City,China(SCKJ-JYRC-2022-75)the Natural Science Foundation of Hainan Province,China(322QN398).
文摘Plant pathogens secrete various cell wall-degrading enzymes that compromise host cell wall integrity and facilitate pathogen invasion.This study identified VdGH7a,a glycoside hydrolase family 7(GH7)cellobiohydrolase from Verticillium dahliae,which demonstrated hydrolytic activity against 1,4-β-glucan.Notably,VdGH7a induced cell death in Nicotiana benthamiana when signal peptides were present,though this effect was inhibited by the carbohydrate-binding type-1(CBM1)protein domain.The deletion of VdGH7a substantially reduced V.dahliae pathogenicity in cotton plants,as demonstrated by the mutants’inability to penetrate cellophane membrane.These knockout mutants also exhibited reduced carbon source utilization capacity and increased sensitivity to osmotic and cell wall stresses.Through yeast two-hybrid screening,bi-molecular fluorescence complementation(BiFC),and luciferase complementation imaging(LCI),we identified that VdGH7a interacts with an osmotin-like protein(GhOLP1)in cotton.Virus-induced gene silencing of GhOLP1 resulted in decreased salicylic acid(SA)content and reduced resistance to V.dahliae in cotton,while heterologous overexpression of GhOLP1 in Arabidopsis enhanced both resistance and SA signaling pathway gene expression.These results reveal a virulence mechanism wherein the secreted protein VdGH7a from V.dahliae interacts with GhOLP1 to activate host immunity and contribute significantly to plant resistance against V.dahliae.
基金National Science and Technology Infrastructure of China,Grant/Award Number:National Pathogen Resource Center-NPRC-32National Key Research and Development Program of China,Grant/Award Number:2023YFF0724800CAMS Innovation Fund for Medical Sciences,Grant/Award Number:2021-I2M-1-035。
文摘Background:New variants of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)continue to drive global epidemics and pose significant health risks.The pathogenicity of these variants evolves under immune pressure and host factors.Understanding these changes is crucial for epidemic control and variant research.Methods:Human angiotensin-converting enzyme 2(hACE2)transgenic mice were in-tranasally challenged with the original strain WH-09 and the variants Delta,Beta,and Omicron BA.1,while BALB/c mice were challenged with Omicron subvariants BA.5,BF.7,and XBB.1.To compare the pathogenicity differences among variants,we con-ducted a comprehensive analysis that included clinical symptom observation,meas-urement of viral loads in the trachea and lungs,evaluation of pulmonary pathology,analysis of immune cell infiltration,and quantification of cytokine levels.Results:In hACE2 mice,the Beta variant caused significant weight loss,severe lung inflammation,increased inflammatory and chemotactic factor secretion,greater mac-rophage and neutrophil infiltration in the lungs,and higher viral loads with prolonged shedding duration.In contrast,BA.1 showed a significant reduction in pathogenicity.The BA.5,BF.7,and XBB.1 variants were less pathogenic than the WH-09,Beta,and Delta variants when infected in BALB/c mice.This was evidenced by reduced weight loss,diminished pulmonary pathology,decreased secretion of inflammatory factors and chemokines,reduced macrophage and neutrophil infiltration,as well as lower viral loads in both the trachea and lungs.Conclusion:In hACE2 mice,the Omicron variant demonstrated the lowest pathogenic-ity,while the Beta variant exhibited the highest.Pathogenicity of the Delta variant was comparable to the original WH-09 strain.Among BALB/c mice,Omicron subvari-ants BA.5,BF.7,and XBB.1 showed no statistically significant differences in virulence.
文摘In Candida species,the endoplasmic reticulum(ER)stress response—regulated by the unfolded protein response(UPR)—serves as a critical adaptive mechanism affecting both pathogenicity and antifungal resistance.This review aims to synthesize current knowledge on ER stress pathways in Candida glabrata and Candida albicans,highlighting their species-specific adaptations and therapeutic implications.We systematically analyzed peer-reviewed literature on ER stress mechanisms in Candida,focusing on comparative studies of UPR signaling.Emphasis was placed on C.glabrata’s inositol-requiring enzyme 1(IRE1)-dependent Regulated IRE1-Dependent Decay(RIDD)pathway and C.albicans’IRE1/HAC1 and calcium-mediated pathways.Connections to virulence and drug resistance were evaluated through genetic,transcriptomic,and phenotypic evidence.Candida species employ divergent UPR strategies:C.glabrata mitigates ER stress primarily via RIDD-mediated mRNA decay to reduce protein load,while C.albicans enhances folding capacity through HAC1 splicing and calcium homeostasis.These adaptations promote survival in hostile host environments(e.g.,oxidative stress,immune attacks)and are linked to resistance against azoles and echinocandins.Pharmacological disruption of UPR components(e.g.,IRE1 inhibitors)sensitizes Candida to antifungals in experimental models.ER stress response pathways are promising targets for antifungal drug development.Understanding species-specific UPR mechanisms in Candida could guide novel therapies to overcome resistance and improve treatment outcomes.
基金funded by the National Key Research and Development Program of China(Grant No.2021YFD1800201).
文摘The H4 subtype of avian influenza virus(AIV)is prevalent worldwide,but only receives little attention due to its low pathogenicity in poultry.Consequently,it remains largely unclear whether H4 AIVs pose a potential threat to the poultry industry and public health.During the period from 2011 to 2022,we conducted an active surveillance programm.A total of 154,762 swab samples were collected across various provinces,and 427 H4 viruses were detected,resulting in a positivity rate of 0.28%.All H4 viruses were isolated from poultry,primarily from ducks in live poultry markets.We further investigated the genetic evolutionary characteristics and pathogenicity of 20 H4Nx viruses isolated in our program.Phylogenetic analysis revealed that the 20 H4Nx viruses belonged to the Eurasian lineage and exhibited significant genetic diversity,with 19 distinct genotypes identified.Molecular characterization indicated that these viruses were low-pathogenicity AIVs with limited binding affinity to human receptors,yet they contained mutations associated with enhanced viral replication and pathogenicity in mammals.Pathogenicity tests conducted in ducks demonstrated that H4 viruses were weakly pathogenic,exhibiting limited replication and transmission capabilities.However,some viruses were able to replicate effectively in mice and induce weight loss.For instance,DK/AH/AG61/11(H4N6)can replicate efficiently in MDCK cells,indicating a potential threat to mammals.These findings underscore the importance of ongoing surveillance of H4 AIVs to better understand their evolution and transmission dynamics and to prevent potential public health risks.
基金supported by the National Natural Science Foundation of China Young Scholars Project(31902242)the Agricultural Science and Technology Innovation Program(ASTIP)of Chinese Academy of Agricultural Sciences(2017–2020)。
文摘Avian pathogenic Escherichia coli(APEC)belonging to extraintestinal pathogenic E.coli(ExPEC)can cause severe infections in extraintestinal tissues in birds and humans,such as the lungs and blood.MprA(microcin production regulation,locus A,herein renamed AbsR,a blood survival regulator),a member of the MarR(multiple antibiotic resistance regulator)transcriptional regulator family,governs the expression of capsule biosynthetic genes in human ExPEC and represents a promising druggable target for antimicrobials.However,a deep understanding of the AbsR regulatory mechanism as well as its regulon is lacking.In this study,we present a systems-level analysis of the APEC AbsR regulon using ChIP-Seq(chromatin immunoprecipitation sequencing)and RNA-Seq(RNA sequencing)methods.We found that AbsR directly regulates 99 genes and indirectly regulates 667 genes.Furthermore,we showed that:1)AbsR contributes to antiphagocytotic effects by macrophages and virulence in a mouse model for systemic infection by directly activating the capsular gene cluster;2)AbsR positively impacts biofilm formation via direct regulation of the T2SS(type II secretion system)but plays a marginal role in virulence;and 3)AbsR directly upregulates the acid tolerance signaling system EvgAS to withstand acid stress but is dispensable in ExPEC virulence.Finally,our data indicate that the role of AbsR in virulence gene regulation is relatively conserved in ExPEC strains.Altogether,this study provides a comprehensive analysis of the AbsR regulon and regulatory mechanism,and our data suggest that AbsR likely influences virulence primarily through the control of capsule production.Interestingly,we found that AbsR severely represses the expression of the type I-F CRISPR(clustered regularly interspaced short palindromic repeats)-Cas(CRISPR associated)systems,which could have implications in CRISPR biology and application.
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funded by the National Science Centre,Poland(Project No.:2017/26/D/NZ6/00136).
文摘Pathogenic microorganisms produce numerous metabolites,including volatile organic compounds(VOCs).Monitoring these metabolites in biological matrices(e.g.,urine,blood,or breath)can reveal the presence of specific microorganisms,enabling the early diagnosis of infections and the timely implementation of tar-geted therapy.However,complex matrices only contain trace levels of VOCs,and their constituent com-ponents can hinder determination of these compounds.Therefore,modern analytical techniques enabling the non-invasive identification and precise quantification of microbial VOCs are needed.In this paper,we discuss bacterial VOC analysis under in vitro conditions,in animal models and disease diagnosis in humans,including techniques for offline and online analysis in clinical settings.We also consider the advantages and limitations of novel microextraction techniques used to prepare biological samples for VOC analysis,in addition to reviewing current clinical studies on bacterial volatilomes that address inter-species in-teractions,the kinetics of VOC metabolism,and species-and drug-resistance specificity.
基金financially supported by the National Nature Science Foundation of China (32372077)the Project of Chongqing Science and Technology Commission (CSTB2023NSCQ-MSX0355)the Fundamental Research Funds for the Central Universities (SWU120075)。
文摘The necrotrophic fungus, Sclerotinia sclerotiorum, employs an array of cell wall-degrading enzymes(CWDEs), including cellulase, to dismantle host cell walls. However, the molecular mechanisms through which S. sclerotiorum degrades cellulose remain elusive. Here, we unveil a novel secretory cellobiohydrolase, SsdchA, characterized by a signal peptide and a Glyco_hydro_7(GH7) domain. SsdchA exhibits a robust expression of during early infection stages. Interestingly, colony morphology and growth rates remain unaffected across the wild-type, SsdchA deletion strains and SsdchA overexpression strains on potato dextrose agar(PDA) medium. Nevertheless, the pathogenicity and cellobiohydrolase activity decreased in the SsdchA deletion strains, but enhanced in the SsdchA overexpression strains. Moreover,the heterologous expression of SsdchA in Arabidopsis thaliana leads to reduced cellulose content and heightened susceptibility to S. sclerotiorum. Collectively, our data underscore the pivotal role of the novel cellobiohydrolase SsdchA in the pathogenicity of S. sclerotiorum.
基金supported by the grants from the Hainan Provincial Natural Science Foundation,China(321QN190 and 321CXTD437)the National Natural Science Foundation of China(32202246 and 32160614)+1 种基金the Open Project Program of Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests,China(MIMCP-202102)the Scientific Research Foundation of Hainan University,China(KYQD(ZR)-21042 and KYQD(ZR)-20080)。
文摘Litchi downy blight,caused by the plant pathogenic oomycete Peronophythora litchii,is one of the most devastating diseases on litchi and resulted in huge economic losses.Autophagy plays an essential role in the development and pathogenicity of the filamentous fungi.However,the function of autophagy in oomycetes remain elusive.Here,an autophagy-related protein Atg3 homolog PlAtg3 was identified and characterized in P.litchii.The absence of PlATG3 through the CRISPR/Cas9 gene replacement strategy compromised vegetative growth and sexual/asexual development.Cytological analyses revealed that the deletion of PlATG3 impaired autophagosome formation with monodansylcadaverine(MDC)staining and significantly disrupted zoospore release due to defects of sporangial cleavage with FM4-64 staining.Atg8 is considered to be an autophagy marker protein in various species.Western blot analysis indicated that PlAtg3 is involved in degradation of PlAtg8-PE.Interestingly,PlAtg3 was unable to interact with PlAtg8 in yeast two hybrid(Y2H)assays,possibly due to the absence of the Atg8 family interacting motif(AIM)in PlAtg3.Furthermore,pathogenicity assays revealed that the deletion of PlATG3 considerably reduced the virulence of P.litchii.Taken together,our data reveal that PlAtg3 plays an important role in radial growth,asexual/sexual development,sporangial cleavage and zoospore release,autophagosome formation,and pathogenicity in P.litchii.This study contributes to a better understanding of the pathogenicity mechanisms of P.litchii and provides insights for the development of more effective strategies to control oomycete diseases.
基金This work was supported by the National Key Research andDevelopment Programof China(2021YFD1800200 and2021YFC2301700).
文摘The stamping-out strategy has been used to control highly pathogenic avian influenza viruses in many countries,driven by the belief that vaccination would not be successful against such viruses and fears that avian influenza virus in vaccinated birds would evolve more rapidly and pose a greater risk to humans.In this review,we summarize the successes in controlling highly pathogenic avian influenza in China and make suggestions regarding the requirements for vaccine selection and effectiveness.In addition,we present evidence that vaccination of poultry not only eliminates human infection with avian influenza virus,but also significantly reduces and abolishes some harmful characteristics of avian influenza virus.
文摘AIM:To investigate the types of bacteria in patients with eye infections in Suzhou and their drug resistance to commonly used antibacterial drugs.METHODS:The clinical data of 155 patients were retrospectively collected in this study,and the pathogenic bacteria species and drug resistance of each pathogenic bacteria were analyzed.RESULTS:Among the 155 patients(age from 12 to 87 years old,with an average age of 57,99 males and 56 females)with eye infections(160 eyes:74 in the left eye,76 in the right eye and 5 in both eyes,all of which were exogenous),71(45.81%)strains were gram-positive bacteria,23(14.84%)strains were gram-negative bacteria and 61(39.35%)strains were fungi.Gram-positive bacteria were highly resistant to penicillin and erythromycin(78.87%and 46.48%respectively),but least resistant to vancomycin at 0.Gram-negative bacteria were highly resistant to cefoxitin and compound sulfamethoxazole(100%and 95.65%respectively),but least resistant to meropenem at 0.Comparison of the resistance of gram-positive and gram-negative bacteria to some drugs revealed statistically significant differences(P<0.05)in the resistance of both to cefoxitin,cotrimoxazole,levofloxacin,cefuroxime,ceftriaxone and ceftazidime,and both had higher rates of resistance to gram-negative bacteria than to gram-positive bacteria.The distribution of bacterial infection strains showed that Staphylococcus epidermidis was the most common strain in the conjunctiva,cornea,aqueous humor or vitreous body and other eye parts.Besides,Fusarium and Pseudomonas aeruginosa were also among the most common strains of conjunctival and corneal infections.CONCLUSION:Gram-positive bacteria are the dominant bacteria in eye infections,followed by gram-negative bacteria and fungi.Considering the resistance of gramnegative bacteria to multiple drugs,monitoring of bacteria should be strengthened in eye bacterial infections for effective prevention and control to reduce complications caused by eye infections.
基金the Guangdong Major Project of Basic and Applied Basic Research(2020B0301030007).
文摘Bordetella bronchiseptica(Bb)is recognized as a leading cause of respiratory diseases in dogs and cats.However,epidemiological data on Bb in dogs and cats in China are still limited,and there is no commercially available vaccine.Live vaccines containing Bb that are widely used abroad are generally efective but can establish latency and potentially reactivate to cause illness in some immunodefcient vaccinated recipients,raising safety concerns.In this study,34 canine-derived and two feline-derived Bb strains were isolated from 1809 canine and 113 feline nasopharyngeal swab samples collected from eight provinces in China from 2021 to 2023.The PCR results showed that the percentage of positive Bb was 22.94%(441/1922),and more than 90%of the Bb isolates had four virulence factor-encoding genes(VFGs),namely,fhaB,prn,betA and dnt.All the isolated strains displayed a multidrug-resistant phenotype.The virulence of 10 Bb strains isolated from dogs with respiratory symptoms was tested in mice,and we found that eight isolates were highly virulent.Furthermore,the eight Bb isolates with high virulence were inactivated and intramuscularly injected into mice,and three Bb strains(WH1218,WH1203 and WH1224)with the best protective efcacy were selected.Dogs immunized with these three strains exhibited strong protection against challenge with the Bb feld strain WH1218.Ultimately,the WH1218 strain with the greatest protection in dogs was selected as the vaccine candidate.Dogs and cats that received a vaccine containing 109 CFU of the inactivated WH1218 strain showed complete protection against challenge with the Bb feld strain WH1218.This study revealed that Bb is an important pathogen that causes respiratory diseases in domestic dogs and cats in China,and all the isolates exhibited multidrug resistance.The present work contributes to the current understanding of the prevalence,antimicrobial resistance,and virulence genes of Bb in domestic dogs and cats.Additionally,our results suggest that the WH1218 strain is a promising candidate safe and efcacious inactivated Bb vaccine.
基金supported by the National Natural Science Foundation of China(82200725)the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202002)+4 种基金the Fundamental Research Funds for the Central Universities(226-2023-00114,226-2022-00226,and 226-2023-00059)the Key Program of National Natural Science Foundation of China(81930016)the Key Research and Development Program of China(2021YFA1100500)the Major Research Plan of the National Natural Science Foundation of China(92159202)the Ningbo Top Medical and Health Research Program(2022030309).
文摘Liver transplantation(LT)is the standard therapy for individuals afflicted with end-stage liver disease.Despite notable advancements in LT technology,the incidence of early allograft dysfunction(EAD)remains a critical concern,exacerbating the current organ shortage and detrimentally affecting the prognosis of recipients.Unfortunately,the perplexing hepatic heterogeneity has impeded characterization of the cellular traits and molecular events that contribute to EAD.Herein,we constructed a pioneering single-cell transcriptomic landscape of human transplanted livers derived from non-EAD and EAD patients,with 12 liver samples collected from 7 donors during the cold perfusion and portal reperfusion stages.Comparison of the 75231 cells of non-EAD and EAD patients revealed an EAD-associated immune niche comprising mucosal-associated invariant T cells,granzyme B^(+)(GZMB^(+))granzyme K^(+)(GZMK^(+))natural killer cells,and S100 calcium binding protein A12^(+)(S100A12^(+))neutrophils.Moreover,we verified this immune niche and its association with EAD occurrence in two independent cohorts.Our findings elucidate the cellular characteristics of transplanted livers and the EAD-associated pathogenic immune niche at the single-cell level,thus,offering valuable insights into EAD onset.
基金Supported by Shenzhen Science and Technology Program,Shenzhen,China(No.JCYJ20200109145001814,No.SGDX20211123120001001)the National Natural Science Foundation of China(No.81970790)Sanming Project of Medicine in Shenzhen(No.SZSM202011015).
文摘AIM:To describe the clinical,electrophysiological,and genetic features of an unusual case with an RDH12 homozygous pathogenic variant and reviewed the characteristics of the patients reported with the same variant.METHODS:The patient underwent a complete ophthalmologic examination including best-corrected visual acuity,anterior segment and dilated fundus,visual field,spectral-domain optical coherence tomography(OCT)and electroretinogram(ERG).The retinal disease panel genes were sequenced through chip capture high-throughput sequencing and Sanger sequencing was used to confirm the result.Then we reviewed the characteristics of the patients reported with the same variant.RESULTS:A 30-year male presented with severe early retinal degeneration who complained night blindness,decreased visual acuity,vitreous floaters and amaurosis fugax.The best corrected vision was 0.04 OD and 0.12 OS,respectively.The fundus photo and OCT showed bilateral macular atrophy but larger areas of macular atrophy in the left eye.Autofluorescence shows bilateral symmetrical hypo-autofluorescence.ERG revealed that the amplitudes of a-and b-wave were severely decreased.Multifocal ERG showed decreased amplitudes in the local macular area.A homozygous missense variant c.146C>T(chr14:68191267)was found.The clinical characteristics of a total of 13 patients reported with the same pathologic variant varied.CONCLUSION:An unusual patient with a homozygous pathogenic variant in the c.146C>T of RDH12 which causes late-onset and asymmetric retinal degeneration are reported.The clinical manifestations of the patient with multimodal retinal imaging and functional examinations have enriched our understanding of this disease.
