Peripheral nerve injury causes severe neuroinflammation and has become a global medical challenge.Previous research has demonstrated that porcine decellularized nerve matrix hydrogel exhibits excellent biological prop...Peripheral nerve injury causes severe neuroinflammation and has become a global medical challenge.Previous research has demonstrated that porcine decellularized nerve matrix hydrogel exhibits excellent biological properties and tissue specificity,highlighting its potential as a biomedical material for the repair of severe peripheral nerve injury;however,its role in modulating neuroinflammation post-peripheral nerve injury remains unknown.Here,we aimed to characterize the anti-inflammatory properties of porcine decellularized nerve matrix hydrogel and their underlying molecular mechanisms.Using peripheral nerve injury model rats treated with porcine decellularized nerve matrix hydrogel,we evaluated structural and functional recovery,macrophage phenotype alteration,specific cytokine expression,and changes in related signaling molecules in vivo.Similar parameters were evaluated in vitro using monocyte/macrophage cell lines stimulated with lipopolysaccharide and cultured on porcine decellularized nerve matrix hydrogel-coated plates in complete medium.These comprehensive analyses revealed that porcine decellularized nerve matrix hydrogel attenuated the activation of excessive inflammation at the early stage of peripheral nerve injury and increased the proportion of the M2 subtype in monocytes/macrophages.Additionally,porcine decellularized nerve matrix hydrogel negatively regulated the Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-κB axis both in vivo and in vitro.Our findings suggest that the efficacious anti-inflammatory properties of porcine decellularized nerve matrix hydrogel induce M2 macrophage polarization via suppression of the Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-κB pathway,providing new insights into the therapeutic mechanism of porcine decellularized nerve matrix hydrogel in peripheral nerve injury.展开更多
Porcine circovirus type 3(PCV3)was initially identified in 2016 in pigs exhibiting unexplained cardiac and multi-organ inflammation in the USA(Palinski et al.2017).PCV3 has subsequently been identified in numerous cou...Porcine circovirus type 3(PCV3)was initially identified in 2016 in pigs exhibiting unexplained cardiac and multi-organ inflammation in the USA(Palinski et al.2017).PCV3 has subsequently been identified in numerous countries,including China,Brazil,Italy,and others,demonstrating widespread viral dissemination(Tan et al.2021).Notably,recent investigations have revealed PCV3 infection across multiple species,including pigs,cattle,dogs,wild boars,chamois,roe deer,and others(Tan et al.2021).This evidence suggests potential viral propagation beyond its primary host(pigs).展开更多
Background:The efficacy of balloon angioplasty for treating peripheral artery disease is influenced by various factors,some of them not yet totally understood.This study aimed to evaluate the role of elastin content i...Background:The efficacy of balloon angioplasty for treating peripheral artery disease is influenced by various factors,some of them not yet totally understood.This study aimed to evaluate the role of elastin content in vascular responses 28 days postangioplasty using uncoated and paclitaxel-coated balloons with the same platform in femoral arteries of a healthy porcine model.Methods:Eight animals underwent balloon angioplasty on the external and internal branches of femoral arteries.Histopathologic evaluation was conducted at follow-up to assess the elastin content,vascular damage,morphological features,and neointimal formation.Results:The elastin content was significantly higher in the external than in the internal femoral artery(p=0.0014).After balloon angioplasty,it was inversely correlated with vascular injury score(ρ=−0.4510,p=0.0096),neointimal inflammation(ρ=−0.3352,p=0.0607),transmural(ρ=−0.4474,p=0.0103)and circumferential(ρ=−0.4591,p=0.0082)smooth muscle cell loss,presence of proteoglycans(ρ=−0.5172,p=0.0024),fibrin deposition(ρ=−0.3496,p=0.0499),and adventitial fibrosis(ρ=−0.6229,p=0.0002).Neointimal formation inhibition with paclitaxel was evident only in arteries with disruption of the internal elastic lamina,with a significant smaller neointimal area in arteries treated with paclitaxel-coated balloons compared to uncoated balloons(median[Q1–Q3]:10.25[7.49–15.64]vs.24.44[18.96–30.52],p=0.0434).Conclusions:Elastin content varies between branches of the femoral artery and significantly influences the integrity of the internal elastic lamina,the vessel's adaptive response,and paclitaxel efficacy after balloon angioplasty.展开更多
Porcine deltacoronavirus(PDCoV)is a swine enteropathogenic CoV that causes severe vomiting,diarrhea and dehydration in suckling piglets,leading to economic losses in the swine industry.There is a great need for a conv...Porcine deltacoronavirus(PDCoV)is a swine enteropathogenic CoV that causes severe vomiting,diarrhea and dehydration in suckling piglets,leading to economic losses in the swine industry.There is a great need for a convenient method to detect circulating antibodies and help in accurate diagnosis and disease control.Previously,we demonstrated that a unique PDCoV accessory protein,NS6,is expressed during PDCoV infection in pigs and is incorporated into PDCoV virions;thus,we deduced that NS6 is likely an immunogenic target that can be used for the diagnosis of PDCoV infection.In this study,we first confirmed that NS6 is immunogenic in PDCoV-infected pigs by perform-ing a serum western blot.Furthermore,we developed a novel NS6-based indirect enzyme-linked immunosorbent assay(iELISA)method and compared it to an established S1-based iELISA for the survey of anti-PDCoV IgG or IgA in pigs of different ages in China.The NS6-iELISA has high specificity for the detection of IgG antibodies and no cross-reactivity with other porcine enteric CoVs(transmissible gastroenteritis coronavirus,porcine epidemic diarrhea virus,or swine acute diarrhea syndrome coronavirus).This NS6 serology-based method has great sensitivity and good repeatability,making it a new and cost-saving option for the rapid diagnosis and immunosurveillance of PDCoV,which may also be important for the prevention and control of deltacoronavirus-related infection in pigs and other animals.展开更多
HighlightsA novel conjugative plasmid pHJ90-cfr carrying the multiresistance gene cfr was characterized in Proteus vulgaris.A new IS5-family member,ISPmi4,was identified for the first time.Both plasmids and ICEs were ...HighlightsA novel conjugative plasmid pHJ90-cfr carrying the multiresistance gene cfr was characterized in Proteus vulgaris.A new IS5-family member,ISPmi4,was identified for the first time.Both plasmids and ICEs were vital mobile genetic elements for horizontal transmission of cfr gene in Proteus species.展开更多
The host intestinal microbiota has emerged as the third element in the interactions between hosts and enteric viruses,and potentially affects the infection processes of enteric viruses.However,the interaction of porci...The host intestinal microbiota has emerged as the third element in the interactions between hosts and enteric viruses,and potentially affects the infection processes of enteric viruses.However,the interaction of porcine enteric coronavirus with intestinal microorganisms during infection remains unclear.In this study,we used 16S-rRNA-based Illumina NovaSeq high-throughput sequencing to identify the changes in the intestinal microbiota of piglets mediated by porcine epidemic diarrhea virus(PEDV)infection and the effects of the alterations in intestinal bacteria on PEDV infection and its molecular mechanisms.The intestinal microbiota of PEDV-infected piglets had significantly less diversity than the healthy group and different bacterial community characteristics.Among the altered intestinal bacteria,the relative abundance of Clostridium perfringens was significantly increased in the PEDV-infected group.A strain of C.perfringens type A,named DQ21,was successfully isolated from the intestines of healthy piglets.The metabolites of swine C.perfringens type A strain DQ21 significantly enhanced PEDV replication in porcine intestinal epithelial cell clone J2(IPEC-J2)cells,and PEDV infection and pathogenicity in suckling piglets.Palmitic acid(PA)was identified as one of those metabolites with metabolomic technology,and significantly enhanced PEDV replication in IPEC-J2 cells and PEDV infection and pathogenicity in suckling piglets.PA also increased the neutralizing antibody titer in the immune sera of mice.Furthermore,PA mediated the palmitoylation of the PEDV S protein,which improved virion stability and membrane fusion,thereby enhancing viral infection.Overall,our study demonstrates a novel mechanism of PEDV infection,with implications for PEDV pathogenicity.展开更多
Dear Editor,Porcine astrovirus(PAstV)is a swine enteric virus which can cause diarrhea and vomiting in pigs,particularly in neonatal piglets(Fang et al.,2019;Su etal.,2020).PAstVwasfirstdetectedby electron microscopyi...Dear Editor,Porcine astrovirus(PAstV)is a swine enteric virus which can cause diarrhea and vomiting in pigs,particularly in neonatal piglets(Fang et al.,2019;Su etal.,2020).PAstVwasfirstdetectedby electron microscopyinthe fecal samples from diarrheal piglets in 1980(Bridger,1980).Based on viral genomes,PAstV is divided into five distinct genotypes(PAstV1-PAstV5)(Fang et al.,2019;Mi et al.,2020;Su et al.,2020).The PAstV5 was the main genotype prevalent in China.Xiao reported a positivity rate of 24.8%for PAstV5 as the predominant genotype in Hunan Province(Xiao et al.,2017).Cai reported a positive rate of 7.5%for PAstV5 in Sichuan Province(Cai et al.,2016).A study in Yunnan Province revealed a total infection rate of 39.9%for PAstV2 and PAstV5(Ren et al.,2022).The PAstV genome open read frame 2(ORF2)encodes for a~90 kDa structural polyprotein called capsid protein(Cap)(Arias and DuBois,2017).The region including amino acids(aa)1-415(N-terminus)of the Cap ishighly conserved,and the region starting at aa 416(C-terminus)is extremely variable(Ren et al.,2022).Moreover,the Cap can bind to host cell receptors and is the main target of neutralizing antibodies(Ricemeyer et al.,2022).展开更多
The cross-species infection of coronaviruses has resulted in several major epidemics since 2003.Porcine epidemic diarrhea virus(PEDV)is a devastating swine enteric coronavirus,which targets pigs as the only natural re...The cross-species infection of coronaviruses has resulted in several major epidemics since 2003.Porcine epidemic diarrhea virus(PEDV)is a devastating swine enteric coronavirus,which targets pigs as the only natural reservoir.In this study,the nucleic acid of PEDV was detected in rat fecal samples collected from pig farms.Further animal tests showed that PEDV can cause systemic infections in neonatal mice and rats via intracranial inoculation.The brain,lung,intestine and spleen were all targets for PEDV in rodents in contrast to the intestine being targeted in pigs.Morbidity and mortality vary via different infection routes.PEDV was also detectable in feces after infection,suggesting that the infected rodents were potential infectious sources.Moreover,the cerebral tropism of PEDV was verified in piglets via orally inoculation,which had not been identified before.In conclusion,our findings demonstrate that PEDV could cross the species barrier to infect mice and rats through different routes in experimental settings.Although it is highly devastating to piglets,PEDV changes the target organs and turns to be milder when meeting with new hosts.Based on these findings,more attention should be paid to the potential cross-species infection of PEDV.展开更多
Ischemic heart disease(IHD)remains a leading contributor to cardiovascular disease(CVD)worldwide.Despite advances in diagnostic and therapeutic approaches,translational research demands robust large animal models to b...Ischemic heart disease(IHD)remains a leading contributor to cardiovascular disease(CVD)worldwide.Despite advances in diagnostic and therapeutic approaches,translational research demands robust large animal models to bridge the gap between experimental interventions and clinical application.Among these,porcine models have gained prominence due to their anatomical,physiological,immunological,and genomic similarities to humans.This review provides a comprehensive overview of current methodologies for establishing porcine IHD models,critically assesses emerging rehabilitative strategies,and outlines innovative therapeutic technologies,with the goal of guiding model selection and fostering the development of novel treatment strategies.展开更多
Porcine reproductive and respiratory syndrome(PRRS),a highly infectious immunosuppressive disease caused by porcine reproductive and respiratory syndrome virus(PRRSV),has led to significant economic losses in the glob...Porcine reproductive and respiratory syndrome(PRRS),a highly infectious immunosuppressive disease caused by porcine reproductive and respiratory syndrome virus(PRRSV),has led to significant economic losses in the global swine industry.The complexity of preventing and controlling PRRS,compounded by the limited efficacy of current vaccines,underscores the urgent need to identify antiviral targets and develop effective therapeutics against PRRSV.From the perspective of virus-host interactions,the discovery of target molecules associated with PRRSV resistance offers a promising strategy for future disease management.In this study,we conduct a comprehensive proteomic analysis using data-independent acquisition(DIA)mode to investigate the host response throughout the acute phase of PRRSV infection.This approach provides critical insights into the regulation of host antiviral and immune pathways during acute infection,advancing our theoretical understanding of PRRSV-host interactions and host gene dynamics during this critical phase.Notably,we identified SCARB2,a major lysosomal membrane protein associated with cholesterol metabolism,as a potential regulator of PRRSV replication.These findings offer novel perspectives for the prevention and control of PRRSV,contributing to the development of targeted antiviral strategies.展开更多
Porcine epidemic diarrhea virus(PEDV)infection causes acute watery diarrhea in neonatal piglets,leading to substantial economic losses within the pig farming industry.This study demonstrates that clofazimine(CFZ)signi...Porcine epidemic diarrhea virus(PEDV)infection causes acute watery diarrhea in neonatal piglets,leading to substantial economic losses within the pig farming industry.This study demonstrates that clofazimine(CFZ)significantly inhibits PEDV replication in a dose-dependent manner in vitro,with negligible cytotoxicity.Findings from our time-of-addition assays indicate that CFZ effectively disrupts multiple stages of the viral infection cycle.Using a CoV-RdRp-Gluc reporter system,we evaluated the potency of CFZ against PEDV RNA-dependent RNA polymerase(RdRp),and determined a low IC50 value of 0.1364μM.Molecular docking studies further confirmed that CFZ has high binding affinity at the active sites of the spike protein and RdRp protein in PEDV.Transcriptome analysis of Vero E6 cells,with and without CFZ treatment,revealed a significant change in transcriptional activity at 8 h postinfection(hpi).Moreover,the simultaneous application of CFZ and nucleoside analogs showed enhanced the anti-PEDV effect of CFZ in vitro.Our study underscores the potential of CFZ as a viable therapeutic agent against PEDV.展开更多
Background:In view of the ever-increasing representation of Staphylococcus spp.strains resistant to various antibiotics,the development of in vivo models for evaluation of novel antimicrobials is of utmost importance....Background:In view of the ever-increasing representation of Staphylococcus spp.strains resistant to various antibiotics,the development of in vivo models for evaluation of novel antimicrobials is of utmost importance.Methods:In this article,we describe the development of a fully immunocompetent porcine model of extensive skin and soft tissue damage suitable for testing topical anti-microbial agents that matches the real clinical situation.The model was developed in three consecutive stages with protocols for each stage amended based on the results of the previous one.Results:In the final model,10 excisions of the skin and underlying soft tissue were created in each pig under general anesthesia,with additional incisions to the fascia performed at the base of the defects and immediately inoculated with Staphylococcus aureus suspension.One pig was not inoculated and used as the negative control.Subsequently,the bandages were changed on Days 4,8,11,and 15.At these time points,a filter paper imprint technique(FPIT)was made from each wound for semi-quantitative microbiological evaluation.Tissue samples from the base of the wound together with the adjacent intact tissue of three randomly selected defects of each pig were taken for microbiological,histopathological,and molecular-biological examination.The infection with the inoculated S.aureus strains was sufficient during the whole experiment as confirmed by both FPIT and from tissue samples.The dynamics of the inflammatory markers and clinical signs of infection are also described.Conclusions:A successfully developed porcine model is suitable for in vivo testing of novel short-acting topical antimicrobial agents.展开更多
Porcine epidemic diarrhea(PED),caused by porcine epidemic diarrhea virus(PEDV),can induce 80–100%mortality in newborn piglets;therefore,specific and rapid detection methods are important for the prevention of this vi...Porcine epidemic diarrhea(PED),caused by porcine epidemic diarrhea virus(PEDV),can induce 80–100%mortality in newborn piglets;therefore,specific and rapid detection methods are important for the prevention of this viral infection.In particular,methods for detecting neutralizing antibodies(nAbs)can be used to evaluate the immunization effect of PEDV vaccines.The spike protein of PEDV(PEDV-S)has been universally used as an antigen to develop immunoassays to detect nAbs.Nanobodies(Nbs)offer advantages such as ease of genetic engineering and low production costs,making them promising for diagnostic applications.In this study,PEDV-S was expressed via the baculovirus system and was used as an antigen to immunize Bactrian camels.A total of 10 Nbs against PEDV-S were first screened and expressed as fusion proteins with horseradish peroxidase(HRP)in HEK293T cells.A Nb-HRP fusion protein named PEDV-S-Nb13-HRP was subsequently selected and used as a probe for developing a competitive enzyme-linked immunosorbent assay(cELISA)to detect anti-PEDV nAbs.Optimization assays identified 80 ng/well of PEDV-S as the optimal coating antigen concentration.The optimal dilution of PEDV-S-Nb13-HRP was 1:200,and the optimal serum dilution was 1:10.The cutoff value of cELISA was determined as 28.1%,demonstrating high specificity,repeatability,stability,and good agreement rates with two commercial ELISA kits(93.6%)and a serum neutralization test(96.34%).Additionally,the results of the detection of IgA antibodies in oral and milk samples from sows were in good agreement with those of the IDEXX PEDV IgA kit.These results demonstrate that the cELISA is a reliable and cost-effective method for detecting anti-PEDV nAbs.展开更多
[Objectives]This study was conducted to find a convenient and reliable method for detecting the virus titer of porcine circovirus type 2(PCV2).[Methods]The reaction conditions of the immunoperoxidase monolayer assay(I...[Objectives]This study was conducted to find a convenient and reliable method for detecting the virus titer of porcine circovirus type 2(PCV2).[Methods]The reaction conditions of the immunoperoxidase monolayer assay(IPMA)method for detecting the viral titer of PCV2 were optimized,and the results were compared with those obtained by indirect immunofluorescence assay(IFA).[Results]PCV2-infected cells exhibited brownish-red staining in either the nucleus or cytoplasm when detected by IPMA,and the detection results were largely consistent with IFA detection.[Conclusions]Both IPMA and IFA methods can be effectively used for determination of PCV2 viral titer,providing reliable support for assessing viral content during PCV2 vaccine development and validating virus inactivation efficacy.展开更多
Background Regulating the regional deposition of fat is crucial for improving the carcass characteristics of pigs.The intestine,as an important organ for lipid absorption and homeostasis maintenance,secretes various b...Background Regulating the regional deposition of fat is crucial for improving the carcass characteristics of pigs.The intestine,as an important organ for lipid absorption and homeostasis maintenance,secretes various biological signals that participate in the crosstalk between the intestine and adipose tissue.Extracellular vesicles,as novel extracellular genetic factors that mediate metabolic signal exchange among multiple tissues,have emerged as a hotspot and breakthrough in revealing the mechanisms of physiological homeostasis.However,how extracellular vesicles regulate the intestinal-adipose signaling axis,especially in relation lipid metabolism and deposition is still unclear.Thus,in the current study,intestinal extracellular vesicles from Chinese fat-type piglets of Lantang and typical leantype piglets of Landrace were isolated and identified,and to reveal the regulatory mechanisms of lipid metabolism via intestinal extracellular vesicles in mediating intestinal-adipose crosstalk.Results We isolated and identified intestinal extracellular vesicles from the jejunum of 3-day-old Lantang and Landrace piglets(LT-EVs and LD-EVs)and further investigated their effects on lipid accumulation in porcine primary adipocytes.Compared to LD-EVs,LT-EVs promoted lipid deposition in porcine primary adipocytes,with intestinal-derived miRNAs playing a critical role in the crosstalk between the intestine and adipose tissue.Further analysis of extracellular vesicles-derived miRNA sequencing revealed that miR-30b-5p,enriched in LD-EVs,is involved in the regulation of lipid metabolism.Notably,the enrichment of miR-30b-5p in extracellular vesicles derived from IPEC-J2 cells also influenced lipid metabolism.Mechanistically,the targeted binding of miR-30b-5p and FMO3 may be critical for the extracellular vesicle-mediated regulation of lipid metabolism.Conclusions Our findings suggest that jejunal-derived extracellular vesicles play a critical role in regulating lipid metabolism,and the regulatory effect of extracellular vesicles from obese piglets was higher than that of lean piglets.Furthermore,the different expression of miRNAs,such as miR-30b-5p,in intestinal extracellular vesicles may be the key to determining lipid deposition phenotypes across the two pig breeds.展开更多
Pluripotent stem cells(PSCs)are useful for developmental and translational research because they have the potential to differentiate into all cell types of an adult individual.Pigs are one of the most important domest...Pluripotent stem cells(PSCs)are useful for developmental and translational research because they have the potential to differentiate into all cell types of an adult individual.Pigs are one of the most important domestic ungulates,commonly used for food and as bioreactors.Generating stable pluripotent porcine PSC lines remains challenging.So far,the pluripotency gene network of porcine PSCs is poorly understood.Here we found that TBX3-derived induced pluripotent stem cells(iPSCs)closely resemble porcine 4-cell embryos with the capacity of totipotent-like stem cells(TLSCs).Interestingly,our data suggest that TBX3 facilitates the activation of H3K4me3 methyltransferase,specifically MLL1.Subsequent investigations revealed that the porcine 4-cell specific gene,MCL1,is a key downstream effector of the TBX3-MLL1 axis.Together,our study of the TBX3 regulatory network is helpful in the understanding of the totipotency characteristics of pigs.展开更多
Porcine deltacoronavirus(PDCoV)is an emerging swine enteropathogenic coronavirus that can cause acute diarrhea and vomiting in newborn piglets and poses a potential risk for cross-species transmission.It is necessary ...Porcine deltacoronavirus(PDCoV)is an emerging swine enteropathogenic coronavirus that can cause acute diarrhea and vomiting in newborn piglets and poses a potential risk for cross-species transmission.It is necessary to develop an effective serological diagnostic tool for the surveillance of PDCoV infection and vaccine immunity effects.In this study,we developed a monoclonal antibody-based competitive ELISA(cELISA)that selected the purified recombinant PDCoV nucleocapsid(N)protein as the coating antigen to detect PDCoV antibodies.To evaluate the diagnostic performance of the cELISA,122 swine serum samples(39 positive and 83 negative)were tested and the results were compared with an indirect immunofluorescence assay(IFA)as the reference method.By receiver operating characteristic(ROC)curve analysis,the optimum cutoff value of percent inhibition(PI)was determined to be 26.8%,which showed excellent diagnostic performance,with an area under the curve(AUC)of 0.9919,a diagnostic sensitivity of 97.44%and a diagnostic specificity of 96.34%.Furthermore,there was good agreement between the cELISA and virus neutralization test(VNT)for the detection of PDCoV antibodies,with a coincidence rate of 92.7%,and theκanalysis showed almost perfect agreement(κ=0.851).Overall,the established cELISA showed good diagnostic performance,including sensitivity,specificity and repeatability,and can be used for diagnostic assistance,evaluating the response to vaccination and assessing swine herd immunity.展开更多
[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated f...[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.展开更多
[Objective] The purification and immunocompetence of GPS protein in porcine reproductive and respiratory syndrome (PRRS) were analyzed in this study, which provided basis for establishing the corresponding serological...[Objective] The purification and immunocompetence of GPS protein in porcine reproductive and respiratory syndrome (PRRS) were analyzed in this study, which provided basis for establishing the corresponding serological method. [Method] The recombinant expression plasmid pGEX-6P-5 was transformed into BL21 and expressed after being induced with IPTG. The solubility analysis of expression products was carried out, and then the recombinant protein was purified for SDS-PAGE identification and Western-blot analysis. Finally, the recombinant antigen was used in the immune experiment of guinea pigs. [Result] The target protein content accounted for 30% of the total cells protein content according to the chromatography scanning, and the purity of target protein after purification reached 80%. The purified protein was analyzed by Western-blot and immune experiment of guinea pigs, and the results showed that the expressed protein had good reactionogenicity and immunogenicity. [Conclusion] This study provides materials for further studies on the function between PRRSV ORF5 gene and its editing protein, which also lays a foundation for porcine reproductive and respiratory syndrome virus genetic engineering products.展开更多
[ Objective] The aim was to analyze the reason and epidemic trend of PRRSV, and provide theoretical basis for preventing and controlling PRRS. [Methed]According to the sequence of ATCC VR-2332 strain porcine reproduct...[ Objective] The aim was to analyze the reason and epidemic trend of PRRSV, and provide theoretical basis for preventing and controlling PRRS. [Methed]According to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published by the GenBank, the primers were designed and synthesized. ORF5 gene sequences of seven prevalence strains were amplified by RT-PCR. The sequences of ORF5 genes were analyzed by DNAStar and compared with those of ATCC VR-2332, CH-1 a, B J-4, LV-M96262 and MLV vaccine strains, phylogenetic tree among isolates was analyzed. [Result] Analysis of nucleotide sequence showed that the homology was 88.1% - 98.8%, 89.9% -95.2%, 85.6% -98.7% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, BJ-4, and the homology was 54.7% -56.9% between ORF5 genes and LV. Analysis of amino acid sequence showed that the homology was 88.1% -96.8%, 88.1% - 94.5%, 86.1% -96.5% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, bBJ-4, the homology was 54.7% -56.2% between the ORF5 genes and LV.[ Conclusion] The variation of prevalence strains was great in the ORF5 gene region, the homology of ORF5 gene sequence was higher and genetic relationship was nearer during prevalence strains in the same region, or was far in different regions.展开更多
基金supported by the Shenzhen Hong Kong Joint Funding Project,No.SGDX20230116093645007(to LY)the Shenzhen Science and Technology Innovation Committee International Cooperation Project,No.GJHZ20200731095608025(to LY)+7 种基金Shenzhen Development and Reform Commission’s Intelligent Diagnosis,Treatment and Prevention of Adolescent Spinal Health Public Service Platform,No.S2002Q84500835(to LY)Shenzhen Medical Research Fund,No.B2303005(to LY)Team-based Medical Science Research Program,No.2024YZZ02(to LY)Zhejiang Provincial Natural Science Foundation of China,No.LWQ20H170001(to RL)Basic Research Project of Shenzhen Science and Technology from Shenzhen Science and Technology Innovation Commission,No.JCYJ20210324103010029(to BY)Shenzhen Second People’s Hospital Clinical Research Fund of Guangdong Province High-level Hospital Construction Project,Nos.2023yjlcyj029(to BY),2023yjlcyj021(to LL)Guangdong Basic and Applied Basic Research Foundation,No.2022A1515110679(to LL)China Postdoctoral Science Foundation,No.2022M722203(to GL).
文摘Peripheral nerve injury causes severe neuroinflammation and has become a global medical challenge.Previous research has demonstrated that porcine decellularized nerve matrix hydrogel exhibits excellent biological properties and tissue specificity,highlighting its potential as a biomedical material for the repair of severe peripheral nerve injury;however,its role in modulating neuroinflammation post-peripheral nerve injury remains unknown.Here,we aimed to characterize the anti-inflammatory properties of porcine decellularized nerve matrix hydrogel and their underlying molecular mechanisms.Using peripheral nerve injury model rats treated with porcine decellularized nerve matrix hydrogel,we evaluated structural and functional recovery,macrophage phenotype alteration,specific cytokine expression,and changes in related signaling molecules in vivo.Similar parameters were evaluated in vitro using monocyte/macrophage cell lines stimulated with lipopolysaccharide and cultured on porcine decellularized nerve matrix hydrogel-coated plates in complete medium.These comprehensive analyses revealed that porcine decellularized nerve matrix hydrogel attenuated the activation of excessive inflammation at the early stage of peripheral nerve injury and increased the proportion of the M2 subtype in monocytes/macrophages.Additionally,porcine decellularized nerve matrix hydrogel negatively regulated the Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-κB axis both in vivo and in vitro.Our findings suggest that the efficacious anti-inflammatory properties of porcine decellularized nerve matrix hydrogel induce M2 macrophage polarization via suppression of the Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-κB pathway,providing new insights into the therapeutic mechanism of porcine decellularized nerve matrix hydrogel in peripheral nerve injury.
基金supported by the National Key Research and Development Program of China (2023YFD1800500)the Jiangsu Innovative and Entrepreneurial Talent Team Project,China (JSSCTD202224)+1 种基金the 111 Project D18007the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD),China
文摘Porcine circovirus type 3(PCV3)was initially identified in 2016 in pigs exhibiting unexplained cardiac and multi-organ inflammation in the USA(Palinski et al.2017).PCV3 has subsequently been identified in numerous countries,including China,Brazil,Italy,and others,demonstrating widespread viral dissemination(Tan et al.2021).Notably,recent investigations have revealed PCV3 infection across multiple species,including pigs,cattle,dogs,wild boars,chamois,roe deer,and others(Tan et al.2021).This evidence suggests potential viral propagation beyond its primary host(pigs).
基金iVascular,S.L.U.,Camíde Can Ubach,11–Nave 3,08620 Sant Vicençdels Horts,Barcelona,Spain。
文摘Background:The efficacy of balloon angioplasty for treating peripheral artery disease is influenced by various factors,some of them not yet totally understood.This study aimed to evaluate the role of elastin content in vascular responses 28 days postangioplasty using uncoated and paclitaxel-coated balloons with the same platform in femoral arteries of a healthy porcine model.Methods:Eight animals underwent balloon angioplasty on the external and internal branches of femoral arteries.Histopathologic evaluation was conducted at follow-up to assess the elastin content,vascular damage,morphological features,and neointimal formation.Results:The elastin content was significantly higher in the external than in the internal femoral artery(p=0.0014).After balloon angioplasty,it was inversely correlated with vascular injury score(ρ=−0.4510,p=0.0096),neointimal inflammation(ρ=−0.3352,p=0.0607),transmural(ρ=−0.4474,p=0.0103)and circumferential(ρ=−0.4591,p=0.0082)smooth muscle cell loss,presence of proteoglycans(ρ=−0.5172,p=0.0024),fibrin deposition(ρ=−0.3496,p=0.0499),and adventitial fibrosis(ρ=−0.6229,p=0.0002).Neointimal formation inhibition with paclitaxel was evident only in arteries with disruption of the internal elastic lamina,with a significant smaller neointimal area in arteries treated with paclitaxel-coated balloons compared to uncoated balloons(median[Q1–Q3]:10.25[7.49–15.64]vs.24.44[18.96–30.52],p=0.0434).Conclusions:Elastin content varies between branches of the femoral artery and significantly influences the integrity of the internal elastic lamina,the vessel's adaptive response,and paclitaxel efficacy after balloon angioplasty.
基金supported by the Zhejiang Provincial Natural Science Foundation(LZ22C180002)the Laboratory of Lingnan Modern Agriculture Project(NG2022001)the Zhejiang Provincial Key R&D Program of China(2021C02049)。
文摘Porcine deltacoronavirus(PDCoV)is a swine enteropathogenic CoV that causes severe vomiting,diarrhea and dehydration in suckling piglets,leading to economic losses in the swine industry.There is a great need for a convenient method to detect circulating antibodies and help in accurate diagnosis and disease control.Previously,we demonstrated that a unique PDCoV accessory protein,NS6,is expressed during PDCoV infection in pigs and is incorporated into PDCoV virions;thus,we deduced that NS6 is likely an immunogenic target that can be used for the diagnosis of PDCoV infection.In this study,we first confirmed that NS6 is immunogenic in PDCoV-infected pigs by perform-ing a serum western blot.Furthermore,we developed a novel NS6-based indirect enzyme-linked immunosorbent assay(iELISA)method and compared it to an established S1-based iELISA for the survey of anti-PDCoV IgG or IgA in pigs of different ages in China.The NS6-iELISA has high specificity for the detection of IgG antibodies and no cross-reactivity with other porcine enteric CoVs(transmissible gastroenteritis coronavirus,porcine epidemic diarrhea virus,or swine acute diarrhea syndrome coronavirus).This NS6 serology-based method has great sensitivity and good repeatability,making it a new and cost-saving option for the rapid diagnosis and immunosurveillance of PDCoV,which may also be important for the prevention and control of deltacoronavirus-related infection in pigs and other animals.
基金supported by the National Key Research and Development Program of China(2022YFF0710505)the Central Public-interest Scientific Institution Basal Research Fund,China(1610302022001)。
文摘HighlightsA novel conjugative plasmid pHJ90-cfr carrying the multiresistance gene cfr was characterized in Proteus vulgaris.A new IS5-family member,ISPmi4,was identified for the first time.Both plasmids and ICEs were vital mobile genetic elements for horizontal transmission of cfr gene in Proteus species.
基金supported by the National Natural Science Foundation of China(U23A20236)the Key Research and Development Program,Guidance Projects of Heilongjiang Province,China(GZ20220029)。
文摘The host intestinal microbiota has emerged as the third element in the interactions between hosts and enteric viruses,and potentially affects the infection processes of enteric viruses.However,the interaction of porcine enteric coronavirus with intestinal microorganisms during infection remains unclear.In this study,we used 16S-rRNA-based Illumina NovaSeq high-throughput sequencing to identify the changes in the intestinal microbiota of piglets mediated by porcine epidemic diarrhea virus(PEDV)infection and the effects of the alterations in intestinal bacteria on PEDV infection and its molecular mechanisms.The intestinal microbiota of PEDV-infected piglets had significantly less diversity than the healthy group and different bacterial community characteristics.Among the altered intestinal bacteria,the relative abundance of Clostridium perfringens was significantly increased in the PEDV-infected group.A strain of C.perfringens type A,named DQ21,was successfully isolated from the intestines of healthy piglets.The metabolites of swine C.perfringens type A strain DQ21 significantly enhanced PEDV replication in porcine intestinal epithelial cell clone J2(IPEC-J2)cells,and PEDV infection and pathogenicity in suckling piglets.Palmitic acid(PA)was identified as one of those metabolites with metabolomic technology,and significantly enhanced PEDV replication in IPEC-J2 cells and PEDV infection and pathogenicity in suckling piglets.PA also increased the neutralizing antibody titer in the immune sera of mice.Furthermore,PA mediated the palmitoylation of the PEDV S protein,which improved virion stability and membrane fusion,thereby enhancing viral infection.Overall,our study demonstrates a novel mechanism of PEDV infection,with implications for PEDV pathogenicity.
基金supported by grants from the Henan Province Key Research and Development Special Project(231111113100)the Henan Province Natural Science Foundation Interdisciplinary Innovation Research Group Project(232300421001)the Postgraduate Education Reform and Quality Improvement Project of Henan Province(YJS2023SZ13).
文摘Dear Editor,Porcine astrovirus(PAstV)is a swine enteric virus which can cause diarrhea and vomiting in pigs,particularly in neonatal piglets(Fang et al.,2019;Su etal.,2020).PAstVwasfirstdetectedby electron microscopyinthe fecal samples from diarrheal piglets in 1980(Bridger,1980).Based on viral genomes,PAstV is divided into five distinct genotypes(PAstV1-PAstV5)(Fang et al.,2019;Mi et al.,2020;Su et al.,2020).The PAstV5 was the main genotype prevalent in China.Xiao reported a positivity rate of 24.8%for PAstV5 as the predominant genotype in Hunan Province(Xiao et al.,2017).Cai reported a positive rate of 7.5%for PAstV5 in Sichuan Province(Cai et al.,2016).A study in Yunnan Province revealed a total infection rate of 39.9%for PAstV2 and PAstV5(Ren et al.,2022).The PAstV genome open read frame 2(ORF2)encodes for a~90 kDa structural polyprotein called capsid protein(Cap)(Arias and DuBois,2017).The region including amino acids(aa)1-415(N-terminus)of the Cap ishighly conserved,and the region starting at aa 416(C-terminus)is extremely variable(Ren et al.,2022).Moreover,the Cap can bind to host cell receptors and is the main target of neutralizing antibodies(Ricemeyer et al.,2022).
基金supported by the Basic Research Innovation Group Project of Gansu Province(No.25JRRA434)the National Natural Science Foundation of China(U22A20522).
文摘The cross-species infection of coronaviruses has resulted in several major epidemics since 2003.Porcine epidemic diarrhea virus(PEDV)is a devastating swine enteric coronavirus,which targets pigs as the only natural reservoir.In this study,the nucleic acid of PEDV was detected in rat fecal samples collected from pig farms.Further animal tests showed that PEDV can cause systemic infections in neonatal mice and rats via intracranial inoculation.The brain,lung,intestine and spleen were all targets for PEDV in rodents in contrast to the intestine being targeted in pigs.Morbidity and mortality vary via different infection routes.PEDV was also detectable in feces after infection,suggesting that the infected rodents were potential infectious sources.Moreover,the cerebral tropism of PEDV was verified in piglets via orally inoculation,which had not been identified before.In conclusion,our findings demonstrate that PEDV could cross the species barrier to infect mice and rats through different routes in experimental settings.Although it is highly devastating to piglets,PEDV changes the target organs and turns to be milder when meeting with new hosts.Based on these findings,more attention should be paid to the potential cross-species infection of PEDV.
基金supported by the National Key R&D Program of China(2023YFC3603800,2023YFC3603801)National Natural Science Foundation of China(82202793,82172534,82372574)+3 种基金Natural Science Foundation of Sichuan Province(2023NSFSC1999,2023NSFSC1495)Young and Middle-Aged Leading Talent Cultivation Project of Sichuan University(JH20231160)Noncommunicable Chronic Diseases-National Science and Technology Major Project(2024ZD0526000)1·3·5 Project for Disciplines of Excellence,West China Hospital,Sichuan University(ZYJC21038)。
文摘Ischemic heart disease(IHD)remains a leading contributor to cardiovascular disease(CVD)worldwide.Despite advances in diagnostic and therapeutic approaches,translational research demands robust large animal models to bridge the gap between experimental interventions and clinical application.Among these,porcine models have gained prominence due to their anatomical,physiological,immunological,and genomic similarities to humans.This review provides a comprehensive overview of current methodologies for establishing porcine IHD models,critically assesses emerging rehabilitative strategies,and outlines innovative therapeutic technologies,with the goal of guiding model selection and fostering the development of novel treatment strategies.
基金supported by the National Natural Science Foundation of China(grant no.:3217190296,82102755 and 32302887)Guangdong Basic and Applied Basic Research Foundation(grant no.:2023A1515012623 and 2019B1515210030)China Postdoctoral Science Foundation(grant no.:2021M703739)。
文摘Porcine reproductive and respiratory syndrome(PRRS),a highly infectious immunosuppressive disease caused by porcine reproductive and respiratory syndrome virus(PRRSV),has led to significant economic losses in the global swine industry.The complexity of preventing and controlling PRRS,compounded by the limited efficacy of current vaccines,underscores the urgent need to identify antiviral targets and develop effective therapeutics against PRRSV.From the perspective of virus-host interactions,the discovery of target molecules associated with PRRSV resistance offers a promising strategy for future disease management.In this study,we conduct a comprehensive proteomic analysis using data-independent acquisition(DIA)mode to investigate the host response throughout the acute phase of PRRSV infection.This approach provides critical insights into the regulation of host antiviral and immune pathways during acute infection,advancing our theoretical understanding of PRRSV-host interactions and host gene dynamics during this critical phase.Notably,we identified SCARB2,a major lysosomal membrane protein associated with cholesterol metabolism,as a potential regulator of PRRSV replication.These findings offer novel perspectives for the prevention and control of PRRSV,contributing to the development of targeted antiviral strategies.
基金supported by a grant from the National Key Research and Development Plans of China(No.2021YFD1800401)to Zhe Wangthe National Natural Science Foundation of China(No.32070128)to Zhe Wang+5 种基金the Shanghai Biomedical Science and Technology Support Special Project(No.21S11900200)to Zhe Wangthe National Natural Science Foundation of China(No.31472211)to ZhiBiao Yangthe Natural Science Foundation of Shanghai(No.21ZR1433900)to Zhibiao Yangthe National Key Research and Development Program of China(No.2022YFD1800805)to Yanjun Zhouthe Pinduoduo-China Agricultural University Research Fund(PC2024A01003)to Xiangfeng Wangthe Biomedicine and Technology Supporting Project of the Shanghai Science and Technology Innovation Plan(Grant No.22S11902200,China)to Tao Sun.
文摘Porcine epidemic diarrhea virus(PEDV)infection causes acute watery diarrhea in neonatal piglets,leading to substantial economic losses within the pig farming industry.This study demonstrates that clofazimine(CFZ)significantly inhibits PEDV replication in a dose-dependent manner in vitro,with negligible cytotoxicity.Findings from our time-of-addition assays indicate that CFZ effectively disrupts multiple stages of the viral infection cycle.Using a CoV-RdRp-Gluc reporter system,we evaluated the potency of CFZ against PEDV RNA-dependent RNA polymerase(RdRp),and determined a low IC50 value of 0.1364μM.Molecular docking studies further confirmed that CFZ has high binding affinity at the active sites of the spike protein and RdRp protein in PEDV.Transcriptome analysis of Vero E6 cells,with and without CFZ treatment,revealed a significant change in transcriptional activity at 8 h postinfection(hpi).Moreover,the simultaneous application of CFZ and nucleoside analogs showed enhanced the anti-PEDV effect of CFZ in vitro.Our study underscores the potential of CFZ as a viable therapeutic agent against PEDV.
基金Supported by the Ministry of Health of the Czech Republic,Grant/Award Number:NU22-05-00475 and NV19-05-00214。
文摘Background:In view of the ever-increasing representation of Staphylococcus spp.strains resistant to various antibiotics,the development of in vivo models for evaluation of novel antimicrobials is of utmost importance.Methods:In this article,we describe the development of a fully immunocompetent porcine model of extensive skin and soft tissue damage suitable for testing topical anti-microbial agents that matches the real clinical situation.The model was developed in three consecutive stages with protocols for each stage amended based on the results of the previous one.Results:In the final model,10 excisions of the skin and underlying soft tissue were created in each pig under general anesthesia,with additional incisions to the fascia performed at the base of the defects and immediately inoculated with Staphylococcus aureus suspension.One pig was not inoculated and used as the negative control.Subsequently,the bandages were changed on Days 4,8,11,and 15.At these time points,a filter paper imprint technique(FPIT)was made from each wound for semi-quantitative microbiological evaluation.Tissue samples from the base of the wound together with the adjacent intact tissue of three randomly selected defects of each pig were taken for microbiological,histopathological,and molecular-biological examination.The infection with the inoculated S.aureus strains was sufficient during the whole experiment as confirmed by both FPIT and from tissue samples.The dynamics of the inflammatory markers and clinical signs of infection are also described.Conclusions:A successfully developed porcine model is suitable for in vivo testing of novel short-acting topical antimicrobial agents.
基金funded by grants from the National Key R&D Program of China(2023YFD1800304)the National Natural Science Foundation of China to QZ(32273041)+1 种基金the Natural Science Foundation of Shaanxi Province of China(2022JC-12)the Central Public interest Scientific Institution Basal Research Fund,National Data Center of Animal Health.
文摘Porcine epidemic diarrhea(PED),caused by porcine epidemic diarrhea virus(PEDV),can induce 80–100%mortality in newborn piglets;therefore,specific and rapid detection methods are important for the prevention of this viral infection.In particular,methods for detecting neutralizing antibodies(nAbs)can be used to evaluate the immunization effect of PEDV vaccines.The spike protein of PEDV(PEDV-S)has been universally used as an antigen to develop immunoassays to detect nAbs.Nanobodies(Nbs)offer advantages such as ease of genetic engineering and low production costs,making them promising for diagnostic applications.In this study,PEDV-S was expressed via the baculovirus system and was used as an antigen to immunize Bactrian camels.A total of 10 Nbs against PEDV-S were first screened and expressed as fusion proteins with horseradish peroxidase(HRP)in HEK293T cells.A Nb-HRP fusion protein named PEDV-S-Nb13-HRP was subsequently selected and used as a probe for developing a competitive enzyme-linked immunosorbent assay(cELISA)to detect anti-PEDV nAbs.Optimization assays identified 80 ng/well of PEDV-S as the optimal coating antigen concentration.The optimal dilution of PEDV-S-Nb13-HRP was 1:200,and the optimal serum dilution was 1:10.The cutoff value of cELISA was determined as 28.1%,demonstrating high specificity,repeatability,stability,and good agreement rates with two commercial ELISA kits(93.6%)and a serum neutralization test(96.34%).Additionally,the results of the detection of IgA antibodies in oral and milk samples from sows were in good agreement with those of the IDEXX PEDV IgA kit.These results demonstrate that the cELISA is a reliable and cost-effective method for detecting anti-PEDV nAbs.
基金Supported by General Project of Shandong Provincial Natural Science Foundation(ZR2022MC173)Binzhou Young Science and Technology Rising Star Program(QMX2023004)Binzhou Comprehensive Experimental Station of Shandong Swine Industry Technology System Project(SDAIT-08-13).
文摘[Objectives]This study was conducted to find a convenient and reliable method for detecting the virus titer of porcine circovirus type 2(PCV2).[Methods]The reaction conditions of the immunoperoxidase monolayer assay(IPMA)method for detecting the viral titer of PCV2 were optimized,and the results were compared with those obtained by indirect immunofluorescence assay(IFA).[Results]PCV2-infected cells exhibited brownish-red staining in either the nucleus or cytoplasm when detected by IPMA,and the detection results were largely consistent with IFA detection.[Conclusions]Both IPMA and IFA methods can be effectively used for determination of PCV2 viral titer,providing reliable support for assessing viral content during PCV2 vaccine development and validating virus inactivation efficacy.
基金supported by the Project of Guangdong Provincial Nature Science Foundation(2023A151502511)Biological Breeding-National Science and Technology Major Project(2023ZD04068)the National Natural Science Foundation of China(32072814).
文摘Background Regulating the regional deposition of fat is crucial for improving the carcass characteristics of pigs.The intestine,as an important organ for lipid absorption and homeostasis maintenance,secretes various biological signals that participate in the crosstalk between the intestine and adipose tissue.Extracellular vesicles,as novel extracellular genetic factors that mediate metabolic signal exchange among multiple tissues,have emerged as a hotspot and breakthrough in revealing the mechanisms of physiological homeostasis.However,how extracellular vesicles regulate the intestinal-adipose signaling axis,especially in relation lipid metabolism and deposition is still unclear.Thus,in the current study,intestinal extracellular vesicles from Chinese fat-type piglets of Lantang and typical leantype piglets of Landrace were isolated and identified,and to reveal the regulatory mechanisms of lipid metabolism via intestinal extracellular vesicles in mediating intestinal-adipose crosstalk.Results We isolated and identified intestinal extracellular vesicles from the jejunum of 3-day-old Lantang and Landrace piglets(LT-EVs and LD-EVs)and further investigated their effects on lipid accumulation in porcine primary adipocytes.Compared to LD-EVs,LT-EVs promoted lipid deposition in porcine primary adipocytes,with intestinal-derived miRNAs playing a critical role in the crosstalk between the intestine and adipose tissue.Further analysis of extracellular vesicles-derived miRNA sequencing revealed that miR-30b-5p,enriched in LD-EVs,is involved in the regulation of lipid metabolism.Notably,the enrichment of miR-30b-5p in extracellular vesicles derived from IPEC-J2 cells also influenced lipid metabolism.Mechanistically,the targeted binding of miR-30b-5p and FMO3 may be critical for the extracellular vesicle-mediated regulation of lipid metabolism.Conclusions Our findings suggest that jejunal-derived extracellular vesicles play a critical role in regulating lipid metabolism,and the regulatory effect of extracellular vesicles from obese piglets was higher than that of lean piglets.Furthermore,the different expression of miRNAs,such as miR-30b-5p,in intestinal extracellular vesicles may be the key to determining lipid deposition phenotypes across the two pig breeds.
基金funded by the National Basic Research Program of China(2022YFD1302200,2023YFF1000904,2023ZD0404303 and 2021YFD1200301)the National Natural Science Foundation of China(32072806,32372970 and 32002246)+2 种基金the Program of Shaanxi Province Science and Technology Innovation Team,China(2019TD-036)the Key Technologies Demonstration of Animal Husbandry in Shaanxi Province,China(20221086 and 20230978)the Inner Mongolia Autonomous Region Open Competition Project,China(2022JBGS0025).
文摘Pluripotent stem cells(PSCs)are useful for developmental and translational research because they have the potential to differentiate into all cell types of an adult individual.Pigs are one of the most important domestic ungulates,commonly used for food and as bioreactors.Generating stable pluripotent porcine PSC lines remains challenging.So far,the pluripotency gene network of porcine PSCs is poorly understood.Here we found that TBX3-derived induced pluripotent stem cells(iPSCs)closely resemble porcine 4-cell embryos with the capacity of totipotent-like stem cells(TLSCs).Interestingly,our data suggest that TBX3 facilitates the activation of H3K4me3 methyltransferase,specifically MLL1.Subsequent investigations revealed that the porcine 4-cell specific gene,MCL1,is a key downstream effector of the TBX3-MLL1 axis.Together,our study of the TBX3 regulatory network is helpful in the understanding of the totipotency characteristics of pigs.
基金supported by the National Key Research and Development Program(2023YFD1800501)the National Natural Science Foundation of China(32373030,32202787)+5 种基金the S&T Program of Hebei(21322401D)the Jiangsu Province Natural Sciences Foundation(BK20221432,BK20210158)the Jiangsu Agricultural Science and Technology Innovation Fund(CX(22)3028)the Special Project of Northern Jiangsu(SZ-LYG202109)the Open Fund of Shaoxing Academy of Biomedicine of Zhejiang Sci-Tech University(SXAB202215)the Open Fund of Key Laboratory for Prevention and Control of Avian Influenza and Other Major Poultry Diseases,Ministry of Agriculture and Rural Affairs(YDWS202213).
文摘Porcine deltacoronavirus(PDCoV)is an emerging swine enteropathogenic coronavirus that can cause acute diarrhea and vomiting in newborn piglets and poses a potential risk for cross-species transmission.It is necessary to develop an effective serological diagnostic tool for the surveillance of PDCoV infection and vaccine immunity effects.In this study,we developed a monoclonal antibody-based competitive ELISA(cELISA)that selected the purified recombinant PDCoV nucleocapsid(N)protein as the coating antigen to detect PDCoV antibodies.To evaluate the diagnostic performance of the cELISA,122 swine serum samples(39 positive and 83 negative)were tested and the results were compared with an indirect immunofluorescence assay(IFA)as the reference method.By receiver operating characteristic(ROC)curve analysis,the optimum cutoff value of percent inhibition(PI)was determined to be 26.8%,which showed excellent diagnostic performance,with an area under the curve(AUC)of 0.9919,a diagnostic sensitivity of 97.44%and a diagnostic specificity of 96.34%.Furthermore,there was good agreement between the cELISA and virus neutralization test(VNT)for the detection of PDCoV antibodies,with a coincidence rate of 92.7%,and theκanalysis showed almost perfect agreement(κ=0.851).Overall,the established cELISA showed good diagnostic performance,including sensitivity,specificity and repeatability,and can be used for diagnostic assistance,evaluating the response to vaccination and assessing swine herd immunity.
基金Supported by Fundamental and Advanced Research Projects of Henan Province(152300410076,2015-2017)Key Science and Technology Program of Henan Province(152102110048,2015-2017)~~
文摘[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.
基金Supported by National High-tech Research and Development Plan (863 Project) in the Eleventh Five-Year Plan Period (2006AA10A207)~~
文摘[Objective] The purification and immunocompetence of GPS protein in porcine reproductive and respiratory syndrome (PRRS) were analyzed in this study, which provided basis for establishing the corresponding serological method. [Method] The recombinant expression plasmid pGEX-6P-5 was transformed into BL21 and expressed after being induced with IPTG. The solubility analysis of expression products was carried out, and then the recombinant protein was purified for SDS-PAGE identification and Western-blot analysis. Finally, the recombinant antigen was used in the immune experiment of guinea pigs. [Result] The target protein content accounted for 30% of the total cells protein content according to the chromatography scanning, and the purity of target protein after purification reached 80%. The purified protein was analyzed by Western-blot and immune experiment of guinea pigs, and the results showed that the expressed protein had good reactionogenicity and immunogenicity. [Conclusion] This study provides materials for further studies on the function between PRRSV ORF5 gene and its editing protein, which also lays a foundation for porcine reproductive and respiratory syndrome virus genetic engineering products.
文摘[ Objective] The aim was to analyze the reason and epidemic trend of PRRSV, and provide theoretical basis for preventing and controlling PRRS. [Methed]According to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published by the GenBank, the primers were designed and synthesized. ORF5 gene sequences of seven prevalence strains were amplified by RT-PCR. The sequences of ORF5 genes were analyzed by DNAStar and compared with those of ATCC VR-2332, CH-1 a, B J-4, LV-M96262 and MLV vaccine strains, phylogenetic tree among isolates was analyzed. [Result] Analysis of nucleotide sequence showed that the homology was 88.1% - 98.8%, 89.9% -95.2%, 85.6% -98.7% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, BJ-4, and the homology was 54.7% -56.9% between ORF5 genes and LV. Analysis of amino acid sequence showed that the homology was 88.1% -96.8%, 88.1% - 94.5%, 86.1% -96.5% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, bBJ-4, the homology was 54.7% -56.2% between the ORF5 genes and LV.[ Conclusion] The variation of prevalence strains was great in the ORF5 gene region, the homology of ORF5 gene sequence was higher and genetic relationship was nearer during prevalence strains in the same region, or was far in different regions.
