Background Porcine epidemic diarrhea virus (PEDV) primarily infects intestinal epithelial cells, causing severe diarrhea and even death in piglets, which results in substantial economic losses for the global swine industry. Recent studies have suggested that PEDV may exhibit new tissue tropism, including the spleen. Results This study aimed to investigate the infection of the Pandemic PEDV in the spleen based on animal experiments and perform a transcriptomic analysis of response of the piglets spleen infected PEDV using RNA sequencing (RNAseq). The result demonstrated that PEDV primarily colonized the jejunum, but spleen had been also detected high viral RNA titers. Furthermore, the PEDV-N antigen was detectable in the spleens of infected pigs, no significant pathological changes were observed in the regions where the antigen was present. Gene expression analysis identified a total of 21,695 genes in spleen tissue, with 392 differentially expressed genes (DEGs), including 162 upregulated and 230 downregulated. Gene Ontology (GO) analysis revealed significant enrichment in terms related to immune system processes; and, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that the DEGs were enriched in Complement and coagulation cascades, as well as Viral protein interactions with. To validate the RNAseq results, 20 randomly selected genes and nine representative DEGs associated with PEDV-infection were confirmed using real-time quantitative PCR (RT-qPCR), and the results were consistent with the trends observed in RNAseq. Conclusions In conclusion, the piglets spleen infected PEDV activated the innate immune response, and the spleen expression of genes associated with PEDV-infected had significant changes. These findings provide new insights into the extra-intestinal pathogenic mechanisms of PEDV infection.

Early childhood exposure to bisphenol A (BPA) is closely associated with autism spectrum disorder (ASD), though the precise molecular mechanisms remain unclear. To investigate this, we employed an integrated computational approach combining network toxicology, molecular docking, and molecular dynamics simulation. Potential targets of BPA and ASD-related genes were collected from multiple databases, identifying 57 overlapping targets. Protein-protein interaction network analysis highlighted 16 core targets among them. Gene Ontology and KEGG pathway enrichment analyses indicated these targets are primarily involved in synaptic transmission, GABAergic signaling, and neuroactive ligand-receptor interactions. Molecular docking demonstrated potential binding between BPA and several core targets, including estrogen receptor 1 (ESR1), gamma-aminobutyric acid type A receptor subunit beta-2 (GABRB2), and amyloid precursor protein (APP), with binding energies below -5 kcal/mol. The stability of the BPA-ESR1 complex was further supported through 100 ns molecular dynamics simulation. These results suggest that BPA may contribute to ASD by disrupting neuroendocrine pathways and synaptic function via interactions with key targets such as ESR1.

Background Pseudorabies virus (PRV), a member of the family Herpesviridae, is responsible for significant economic losses in the pig industry and has recently been associated with human viral encephalitis, leading to severe neurological symptoms post-recovery. Despite the widespread impact of PRV, there are currently no approved effective drugs for treating PRV-related diseases in humans or pigs. Therefore, the exploration and discovery of safe and effective drugs for the prevention and treatment of PRV infection is of paramount importance. Purpose The objective of this study is to screen and identify natural compounds with antiviral activity against PRV. Methods First, we used a strain of PRV with green fluorescent protein (PRV-GFP) to screen a natural product chemical library to identify potential antiviral drugs. Next, we assessed the antiviral abilities of salvianolic acid A (SAA) in vitro using virus titer assay, qPCR, and IFA. We investigated the mechanisms of SAA's antiviral activity through viral attachment, internalization, inactivation, and nuclease digestion assay. Finally, we evaluated the efficacy of SAA in inactivating PRV using mice as the experimental subjects. Results This study screened 206 natural compounds for anti-PRV activity in vitro, resulting in the identification of seven potential antiviral agents. Notably, SAA emerged as a promising candidate with significant anti-PRV activity. The mechanism of action may be that SAA can directly inactivate the virus by disrupting viral envelope. In vivo experiments have shown that pre-incubation of SAA and PRV can effectively inhibit the infectivity and pathogenicity of PRV in mice. Conclusion This study offers valuable insights into the antiviral properties of SAA, potentially informing strategies for controlling PRV epidemics and treating related diseases in both humans and animals.

Lung adenocarcinoma (LUAD) is the most common subtypes of NSCLC. However, the therapeutic effects for LUAD are unsatisfactory at current stage, so it is important to find new molecular targets and therapeutic strategies. circRNAs can regulate the expression of target genes by binding to microRNAs (miRNAs) to form competitive endogenous RNAs (ceRNAs). Therefore, we investigated the functions of circR-4225 in the tumor progression of LUAD and its molecular mechanism in this paper. circR-4225 is up-regulated in LUAD tissues. EIF4A3, a member of the eukaryotic translation initiation factor 4A (EIF4A) family, promotes the expression of circR-4225. circR-4225 acts as a molecular sponge to down-regulate miR-507, which promotes the up-regulation of the expression of its target gene-tumor necrosis factor superfamily member 11 (TNFSF11). Knockdown of circR-4225 in the LUAD cell lines can inhibit cell proliferation and viability, and promote apoptosis of the LUAD cell lines, which can be reverted by inhibiting miR-507 or overexpressing TNFSF11. To sum it up, this study demonstrated that circR-4225 was significantly up-regulated in LUAD tissues, and circR-4225 promoted LUAD progression by sponging miR-507 and up-regulating TNFSF11. This study can provide new molecular targets for early diagnosis and treatment of LUAD.

The emergence of new coronaviruses poses a significant threat to animal husbandry and human health. Porcine epidemic diarrhea virus (PEDV) is considered a re-emerging porcine enteric coronavirus, which causes fatal watery diarrhea in piglets. Currently, there are no effective drugs to combat PEDV. Drug repurposing screens have emerged as an attractive strategy to accelerate antiviral drug discovery and development. Here, we screened 206 natural products for antiviral activity using live PEDV infection in Vero cells and identified ten candidate antiviral agents. Among them, Tubercidin, a nucleoside analog derived from Streptomyces tubercidicus, showed promising antiviral activity against PEDV infection. Furthermore, we demonstrated that Tubercidin exhibited significant antiviral activity against both classical and variant PEDV. Time of addition assay showed that Tubercidin displayed a significant inhibitory effect on viral post-entry events but not during other periods. Molecular docking analysis indicated that Tubercidin had better docking efficiency and formed hydrophobic interactions with the active pocket of RNA-dependent RNA polymerase (RdRp) of PEDV and other nidoviruses. Additionally, Tubercidin can effectively suppress other porcine nidoviruses, such as SADS-CoV and PRRSV, demonstrating its broad-spectrum antiviral properties. In summary, our findings provide valuable evidence for the antiviral activity of Tubercidin and offer insights into the development of new strategies for the prevention and treatment of coronavirus infections.

Classical swine fever (CSF) is a World Organisation for Animal Health (formerly the Office International des Epizooties [OIE]) notifiable terrestrial and aquatic animal disease, causing great economic losses to the swine industry worldwide during the past decades. Due to the use of the most effective and safe attenuated live vaccine for CSF prevention, differentiation of infected and vaccinated pigs is vital work, as well as a bottleneck for eradication of CSF. As a notifiable terrestrial and aquatic animal disease listed by World Organisation for Animal Health (formerly the Office International des Epizooties [OIE]), classical swine fever (CSF) has caused great economic losses to the swine industry worldwide during recent decades. Differentiation of infected and vaccinated animals (DIVA) is urgent for eradication of CSF. In this study, a diagnostic platform based on CRISPR/Cas13a was established with the ability to differentiate between classical swine fever virus (CSFV) virulent and vaccine strains. In combination with reverse transcription recombinase-aided amplification (RT-RAA), the detection limit for CSFV synthetic RNA templates reached 3.0 x 10(2) copies/mu L. In addition, with boiling and chemical reduction, heating unextracted diagnostic samples to obliterate nucleases (HUDSON) treatment was introduced to inactivate nucleases and release viral genome, achieving robust pretreatment of tested sample before CRISPR/Cas13a detection without the need to extract viral nucleic acids. HUDSON-RT-RAA-CRISPR/Cas13a can directly detect cell cultures of virulent Shimen strain and vaccine hog cholera lapinized virus (HCLV) strain, with the detection limit of 3.5 x 10(2) copies/mu L and 1.8 x 10(2) copies/mu L, respectively, which was equally sensitive to nested PCR (nPCR) and 100 times more sensitive than antigen enzyme-linked immunosorbent assay (ELISA). Meanwhile, HUDSON-RT-RAA-CRISPR/Cas13a showed no cross-reactivity with bovine viral diarrhea virus (BVDV), atypical porcine pestivirus (APPV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV), African swine fever virus (ASFV), pseudorabies virus (PRV), and porcine circovirus 2 (PCV2), exhibiting good specificity. At last, a total of 50 pig spleen samples with suspected clinical signs were also assayed with HUDSON-RT-RAA-CRISPR/Cas13a, nPCR, and antigen ELISA in parallel. HUDSON-RT-RAA-CRISPR/Cas13a showed 100.0% with nPCR and 82.0% coincident rate with antigen ELISA, respectively. IMPORTANCE Classical swine fever (CSF) is a World Organisation for Animal Health (formerly the Office International des Epizooties [OIE]) notifiable terrestrial and aquatic animal disease, causing great economic losses to the swine industry worldwide during the past decades. Due to the use of the most effective and safe attenuated live vaccine for CSF prevention, differentiation of infected and vaccinated pigs is vital work, as well as a bottleneck for eradication of CSF. Methods with the ability to precisely differentiate classical swine fever virus (CSFV) virulent strains from vaccine strain hog cholera lapinized virus (HCLV) are urgently needed. Combining the high sensitivity of isothermal recombinase-aided amplification (RAA) with the accurate molecular sensing ability of Cas13a, we presented a novel method for CSFV detection without the need to extract viral nucleic acids, which showed great advantage to traditional detection methods for precise differentiation of CSFV virulent strains and vaccine strain, providing a novel powerful tool for CSF eradication.

Antigen proteins, assembled on nanoparticles, can be recognized by antigen-presenting cells effectively to enhance antigen immunogenicity. The ability to simultaneously display multiantigens on the same nanoparticle could have numerous applications but remained technical challenges. Here, we described a method for precise assembly of multiple antigens on nanoparticles with specially designed affinity peptides. First, we designed and screened affinity peptides with high affinity and specificity, which could respectively target the key amino acid residues of classical swine fever virus (CSFV) E2 protein or porcine circovirus type 2 capsid protein (PCV2 Cap) accurately. Then, we conjugated the antigen proteins to poly(lactic acid-glycolic acid) copolymer (PLGA) and Gram-positive enhancer matrix (GEM) nanoparticles through the peptides and perfectly assembled two kinds of multiantigen display nanoparticles with different particle sizes. Subsequently, the immunological properties of the assembled nanoparticles were tested. The results showed that the antigen display nanoparticles could promote the maturation, phagocytosis, and proinflammatory effects of antigen-presenting cells (APCs). Besides, compared with the antigen proteins, multiantigen display nanoparticles could induce much higher levels of antibodies and neutralizing antibodies in mice. This strategy may provide a technical support for the study of protein structure and the research and development of polyvalent vaccines.

金黄色葡萄球菌是一种常见的人兽共患病原菌,在自然界分布广泛,可引起人和动物感染,也可引起细菌性食物中毒或饲料中毒.在多种动物及各种食品中均检出了该菌,且检出率不断上升,并在全球扩散,已成为医院病原感染和牛羊乳腺炎感染的重要病原菌之一,食源性细菌与临床菌株可能通过食物链产生交叉污染,能够互相传播,还可能从动物传给人类,给人类和动物健康带来了一定的威胁.论文对金黄色葡萄球菌的流行、分布、感染、致病性与食品安全等方面进行综述.

目的:对近年来发表的国内医学本科教学中混合式教学效果的研究进行meta分析,评价混合式教学模式在医学本科课程理论教学和实践教学中的效果.方法:检索中国知网、万方数据库、中文科技期刊数据库、中国生物医学文献数据库,筛选符合条件的文献,对文献进行质量评价,提取数据,采用revman 5.3对理论成绩和实验实践成绩进行meta分析.结果:共纳入符合标准的文献22篇,混合式教学组的理论、实验成绩与传统教学组的差异均有统计学意义(P＜0.05),理论及实验成绩的SMD及其95％CI分别为0.89(0.51,1.28)、1.05(0.77,1.34).结论:混合式教学的理论成绩和实验成绩均优于传统教学,混合式教学可以提高医学本科教学的教学效果.

当前,人们对健康养生的认识与需求越来越强,回顾中华五千年历史,茶与中医药有着较为紧密的联系.中药代茶饮即以药代茶,兼具防病、养生之功效.而中药复方白头翁汤,是东汉名医张仲景著名的千年药方,现代药理学研究发现白头翁汤中的主要药物成分包含香豆素类、生物碱类、柠檬苦素类和皂甙类等,这些药物成分能够发挥一定的抗菌、抗炎和修复溃疡、免疫调节和抗腹泻等药理作用.在临床应用上,该方在治疗各种肠炎、细菌性痢疾和泌尿系统感染等疾病中发挥了重要的作用.为此,基于药茶理念对中药复方白头翁汤研究进展进行综述,以供参考.

本文以疫情防控期间医学基础课程《医学微生物学》的网上教学为例,对河南中医药大学基础医学院开展网上教学的实践与教学反思进行总结,旨在为提升医学基础课程《医学微生物学》的教学质量.

Background Porcine circovirus type 2 (PCV2) is the pathogen of porcine circovirus associated diseases (PCVAD) and one of the main pathogens in the global pig industry, which has brought huge economic losses to the pig industry. In recent years, there has been limited research on the prevalence of PCV2 in Henan Province. This study investigated the genotype and evolution of PCV2 in this area. Results We collected 117 clinical samples from different regions of Henan Province from 2015 to 2018. Here, we found that the PCV2 infection rate of PCV2 was 62.4%. Thirty-seven positive clinical samples were selected to amplify the complete genome of PCV2 and were sequenced. Based on the phylogenetic analysis of PCV2 ORF2 and complete genome, it was found that the 37 newly detected strains belonged to PCV2a (3 of 37), PCV2b (21 of 37) and PCV2d (13 of 37), indicating the predominant prevalence of PCV2b and PCV2d strains. In addition, we compared the amino acid sequences and found several amino acid mutation sites among different genotypes. Furthermore, the results of selective pressure analysis showed that there were 5 positive selection sites. Conclusions This study indicated the genetic diversity, molecular epidemiology and evolution of PCV2 genotypes in Henan Province during 2015-2018.

In this study, we describe the use of a M13 phage-displayed random peptide library for screening novel peptide motifs that specifically recognize recombinant PCV2 Cap protein for the first time. The peptide had a sequence of YHDCFSAGFCIG, and the binding affinity was observed in the nanomolar range. In the serum blocking test, the ELISA signal of the peptide was rapidly decreased when the concentration of the PCV2 positive serum was increased.

In order to study the pathogenic mechanism of pseudorabies virus(PRV),PK-15 cells were infected with PRV QBA strain at 0.5 MOI,and cells were harvested and microRNA(miRNA) were extracted at 0,1,2,4,6,8,12,24 hours post infection(hpi).Then the temporal expression of prv-miR-LLT11a was detected by stem-loop real-time quantitative PCR(RT-qPCR).The amplification results of RT-qPCR showed that the melting curve had a single peak and a clear inflection point.RT-qPCR data showed that the expression of prv-miR-LLT11a remarkably significant increased in PK-15 cells after 1 hour infection with PRV QBA strain,then decreased.Its expression level decreased to the lowest level at 6 hpi,and increased significantly after 8 hpi.

Pseudorabies virus (PRV), an alpha herpesvirus can enter the mammalian nervous system, causing Aujezsky's disease. Previous studies have reported an alteration of microRNA (miRNA) expression levels during PRV infections. However, knowledge regarding miRNA response in nervous cells to PRV infection is still unknown. To address this issue, small RNA libraries from infected and uninfected mouse neuroblastoma cells were assessed after Illumina deep sequencing. A total of eight viral miRNA were identified, and ten host miRNAs showed significantly different expression upon PRV infection. Among these, five were analyzed by stem-loop RT-qPCR, which confirmed the above data. Interestingly, these viral miRNAs were mainly found in the large latency transcript region of PRV, and predicted to target a variety of genes, forming a complicated regulatory network. Moreover, ten cellular miRNAs were expressed differently upon PRV infection, including nine upregulated and one downregulated miRNAs. Host targets of these miRNAs obtained by bioinformatics analysis belonged to large signaling networks, mainly encompassing calcium signaling pathway, cAMP signaling pathway, MAPK signaling pathway, and other nervous-associated pathways. These findings further highlighted miRNA features in nervous cells after PRV infection and contributed to unveil the underlying mechanisms of neurotropism as well as the neuropathogenesis of PRV.

本研究确定伐昔洛韦的体外和体内抗猪伪狂犬病病毒作用,确定伐昔洛韦对Vero细胞的最大无毒质量浓度为4 g/L,抑制PRV野毒在Vero细胞中复制的最低有效质量浓度为3 g/L.用1个LD50的PRV对小鼠进行攻毒,在攻毒后48 h给服不同剂量的伐昔洛韦,确定了伐昔洛韦能够为小鼠提供完全保护的最低剂量为3 mg/只.PRV攻毒后,未给服伐昔洛韦组小鼠表现典型的神经症状并全部死亡,而给药组小鼠临床健康并全部存活.与给药组小鼠相比,未给药组小鼠脑、肺组织中的PRV病毒载量明显高于给药组小鼠,且随时间延长一直增加.脑、肺组织呈现PRV感染的特征性病理变化.伐昔洛韦给药组小鼠相应组织中的病毒载量和脾脏T淋巴细胞分泌PRV特异性Th1型细胞因子水平在攻毒后72 h逐渐降低.试验结果显示,伐昔洛韦无论在体内、体外均能够有效抑制PRV的复制.

口蹄疫是由口蹄疫病毒引起的偶蹄动物的一种急性热性高度接触性传染病.本病发病急、传播迅速、发病率高、宿主谱系广、危害大,曾多次在世界发生大流行.世界动物卫生组织(OIE)将其列为为A类传染病之首,我国将其列为一类动物传染病.我国目前主要流行0型和A型口蹄疫.2000年以来欧洲一些国家和我国的近邻蒙古、韩国、日本、俄罗斯、中国香港和台湾又相继暴发口蹄疫,疫情不断扩大,同时,东南亚国家口蹄疫长期流行,这些进一步说明我国控制口蹄疫的艰巨性和复杂性.

To research the role of DNA methylation in PRV latent infection,this study firstly predicted methylation status of the PRV genome,then bisulfite PCR sequencing was used to detect methylation status of IE180 and EP0promoter and the adjacent transcription start site region during acute and latent infection.The result shows that IE180 and EP0promoter and the adjacent transcription start site region during acute and latent infection are methylated in scattered,and the wide methylated region was not detected,consistent with the prediction.Preliminary study shows PRV is less likely to egtab lish latent infection by DNA methylation,which indicates that DNA methylation may not play a key role during PRV latent infection.

The animal model was established by adding different doses of molybdate(Mo)into drinking water as the control group(Mo:0mg/L),Mo Ⅰ group(Mo:12.5mg/L),Mo Ⅱgroup(Mo:25.0mg/L),MoⅢ group(Mo:50.0mg/L)and Mo Ⅳ group(Mo:100.0mg/L).On the2 th week and 5th week,all chickens were immunized by Newcastle disease virus attenuated vaccine.On the 7th week and 10 th week,serum was collected to identify HI antibody titer and antioxidative index.The results showed that the body weight and average weight gain were significantly increased(P0.05)in adding 50mg/L Mo.Mo Addition significantly increased the content of serum TPr and Alb,and the activity of XOD and ACP in chickens.The activity of SOD,GSH-Px,GSH,NO and T-NOS were also increased significantly by adding Mo in a dose-dependent manner.In conclusion,Mo addition promotes the growth of chicken,significantly increased the immune function and antioxidant ability.