Non-coding RNAs are crucial orchestrators in the intricate dance between viruses and host cells, among which the expression and function of enhancer RNAs (eRNAs) during influenza virus infection remain largely unexplored. This study utilized whole transcriptome high-throughput sequencing to investigate the molecular mechanisms underpinning the species-specific regulation of influenza virus replication by the miR-302 cluster-IRFs-IRF1AS axis both in vivo and in vitro. Mechanistically, the CTNNB1-induced miR-302 cluster targeted various interferon regulatory factors (mainly IRF1 and IRF2) with varying affinities and silencing efficiencies, except for miR-302e and miR-302f. Furthermore, miR-302 cluster-IRFs triggered the induction of interferon-induced hub genes and hub lncRNAs defined through weighted gene co-expression network analysis. Importantly, the intricate interplay between IRFs, direct targets of the miR-302 cluster, and IRF1AS, an indirect target, in terms of gene loci and transcriptional regulation reveals a crosstalk in the miR-302 cluster-IRFs-IRF1AS axis. That is, on the one hand, IRF1 and IRF7 bind to the promoter of IRF1AS to promote the transcription of eRNAs. On the other hand, IRF1AS functions as an enhancer cluster that orchestrates and cis-regulates the transcription of IRF1, thereby rapidly amplifying the antiviral immune response initiated by miR-302 cluster-IRFs. In conclusion, we have unveiled a novel regulatory network governed by the miR-302 cluster-IRFs-IRF1AS, offering fresh perspectives on immune regulatory mechanisms.IMPORTANCENon-coding RNAs play a crucial role in regulating the three-dimensional structure of chromatin. They influence gene expression through various mechanisms and thereby contribute to the onset and progression of influenza A virus pathogenicity. Our comprehensive whole transcriptome sequencing analysis reveals a novel finding: the species-specific regulation of influenza virus replication by the miR-302 cluster-IRFs-IRF1AS axis. Our findings indicate that the miR-302 cluster-IRFs axis facilitates the transcription of key hub genes and hub lncRNAs, most of which significantly inhibit influenza virus replication. Notably, the downstream IRF1AS assembles into an enhancer cluster, orchestrating and cis-regulating the transcription of IRF1 to activate the interferon system. This investigation enhances our understanding of the regulatory network underlying viral infections and offers novel insights into immune regulatory mechanisms.

Urbanization carries essential influences to ecosystem of soil bacteria in coastal cities. Comprehending the patterns and drivers of bacterial diversity are essential to understanding how soil ecosystems respond to environmental change. This study aimed to explore how soil bacterial community (SBC) response to distinct urbanization of coastal cities on composition, assembly process and potential function in Guangdong province, south China. 72 samples from 24 sample sites within 3 cities were included in the study. Soil chemical properties were analyzed, and the bacterial community were investigated by high-throughout sequencing. Proteobacteria and Acidobacteria were the main phyla. Assembly processes remained in stochastic processes and co-occurrence network of SBC kept stable, while urbanization altered SBC by influencing the dominant phyla. The indicators of communities in coastal city soils were the genera gamma_proteobacterium and beta_proteobacterium. Urbanized extent was the non-negligible factor which affected soil bacterial community, despite the total carbon was still the most vital. The impact of urbanization on bacterial communities might follow a non-linear pattern. Faprotax function prediction showed different urbanized coastal city soils share similar metabolic potential. Our study improved our understanding of the response of soil bacterial communities to urbanization in subtropical coastal cities and offered a useful strategy to monitor the ecology risk toward the soil under urbanization.

[现状]犊牛腹泻是以腹泻为临床症状的一类影响养牛业发展的常见重要疾病.[问题]该病严重时可影响犊牛的生长发育,甚至会造成死亡.该病病因复杂,且发病率和死亡率较高,对养牛业造成巨大经济损失.[对策]因此,在实际生产中,对犊牛腹泻病因做出准确判断并采取行之有效的防控措施尤为重要.[结论]本文从传染病学的角度分病毒、细菌、寄生虫三个方面介绍了导致犊牛腹泻的常见病原体,并阐述了几种常见病原的诊断方法和综合防控措施,为犊牛腹泻的科学防控提供借鉴参考.

[目的]新疆某牛场的犊牛发生严重腹泻,出现排血便、黄色水样稀便、精神萎靡、食欲不振等症状.为了确定此次发病的病原做了此实验.[方法]无菌采集了20份犊牛粪便样品,从粪便中分离纯化出1株细菌,并对其进行培养特性观察、PCR鉴定、黏附性基因检测、药敏试验和动物致病性试验.[结果]结果显示分离到了一株革兰氏阴性菌,根据形状和大小判断疑似大肠杆菌;PCR鉴定结果显示为大肠杆菌,分离菌株具有F41黏附性基因.该分离菌株对多黏菌素B、阿米卡星、头孢他啶、头孢哌酮等敏感;在致病性试验中,小鼠在15 h内陆续死亡,说明该菌株具有一定致病力.[结论]结果表明,此次牛场犊牛腹泻分离的菌株为产肠毒素性大肠杆菌,携带F41毒力基因,对多黏菌素B、阿米卡星等抗生素敏感,具有一定致病力.

为查明新疆喀什地区某牛场发生犊牛腹泻的原因,采集了8头病死犊牛及2头健康犊牛的粪便进行病原学诊断.对采集到的粪便采用细菌分离培养鉴定、药物敏感性试验,并对引起犊牛腹泻的常见病毒开展RT-PCR检测.结果显示分离出的致病菌为革兰氏阴性杆菌,经过16SrDNA测序表明该菌为致病性大肠埃希氏杆菌,其阳性率100％.药物敏感性试验显示,该致病菌对美罗培南、阿米卡星、阿奇霉素3种抗生素敏感,对其它10种抗生素耐药.牛冠状病毒(Bovine coronavirus,BCoV)RT-PCR检测阳性率为75％,牛病毒性腹泻/粘膜病(Bovine viral diarrhea/mucosal disease,BVD/MD)、牛轮状病毒(Bovine Rotaviruses,BRV)均为阴性.结论为该牛场犊牛腹泻主要由牛冠状病毒和致病性大肠埃希氏杆菌混合感染引起.

Epinephelus coioides is a fish species with high economic value due to its delicious meat, high protein content, and rich fatty acid nutrition. It has become a high-economic fish in southern parts of China and some other Southeast Asian countries. In this study, the myostatin nucleic acid vaccine was constructed and used to immunize E. coioides. The results from body length and weight measurements indicated the myostatin nucleic acid vaccine promoted E. coioides growth performance by increasing muscle fiber size. The results from RT-qPCR analysis showed that myostatin nucleic acid vaccine upregulated the expression of myod, myog and p21 mRNA, downregulated the expression of smad3 and mrf4 mRNA. This preliminary study is the first report that explored the role of myostatin in E. coioides and showed positive effects of autologous nucleic acid vaccine on the muscle growth of E. coioides. Further experiments with increased numbers of animals and different doses are needed for its application to E. coiodes aquaculture production.

[目的]构建多拷贝整合表达禽流感病毒样颗粒的重组巴斯德毕赤酵母,为H5 N1亚型禽流感基因工程疫苗提供基础.[方法]以巴斯德毕赤酵母GS115株18S rRNA基因(rDNA)部分序列,插入pPIC9K载体上XbaⅠ和Bsp1407Ⅰ位点间,构建多拷贝整合表达质粒p8K.再以禽流感病毒H5N1毒株基因组RNA为模板,RT-PCR扩增HA、M和NA基因,分别插入多拷贝载体p8K,构建表达质粒p8K-HA/M/NA.表达质粒分别扩增后线性化,按比例混合,电转化GS115感受态细胞.增菌后,经遗传霉素G418抗性平板筛选、PCR鉴定携带HA、M和NA基因序列的多拷贝整合菌株.阳性菌株增菌、诱导表达、高压均质破碎后,Western-blot检测表达产物.[结果]PCR鉴定阳性的重组菌株,其细胞裂解蛋白,免疫印迹试验可检测到与HA、M1和NA分子量一致的蛋白条带;电子显微镜可观察到大小约为80～120 nm的病毒样颗粒,免疫鸡能产生抗禽流感病毒中和抗体.[结论]重组毕赤酵母能够多拷贝整合、表达、组装禽流感病毒样颗粒.

为了解新疆野生北山羊小反刍兽疫病毒(peste des petits ruminants virus,PPRV)的分子特征,根据GenBank公布的PPRV基因组序列设计并合成引物,采用RT-PCR方法和基因测序技术获得病毒全基因序列,应用分子生物学分析软件,对分离的PPRV毒株进行序列分析.结果显示,本次分离的PPRV毒株(China/XJAKS/2017)属于基因IV型,基因组全长15954 nt,编码6种结构蛋白和2种非结构蛋白;在系统进化上,与国内新疆分离株China/XJBZ/2015、China/XJYL/2013同源率分别高达99.0％、99.6％,与国外的巴基斯坦和塔吉克斯坦分离株亲缘关系最近.结果表明,PPRV已经在家养动物与野生动物之间传播.结果提示,PPRV传入野生动物群为我国消除PPR带来极大困难,需加强我国边疆地区PPR免疫隔离带建设,并对野生动物PPRV分子流行病学特点进行追踪监测.

In this study, we analyzed the complete mitochondrial genome (mitogenome) of Speiredonia retorta, which is a pest and a member of the Lepidoptera order. In total, the S. retorta mitogenome was found to contain 15,652 base pairs encoding 13 protein-coding genes (PCGs), 22 tRNAs, 2 rRNAs, as well as an adenine (A)+thymine (T)-rich region. These findings were consistent with the mitogenome composition of other lepidopterans, as we identified all 13 PCGs beginning at ATN codons. We also found that 11 PCGs terminated with canonical stop codons, whereas cox2 and nad4 exhibited incomplete termination codons. By analyzing the mitogenome of S. retorta using Bayesian inference (BI) and maximum likelihood (ML) models, we were able to further confirm that this species is a member of the Erebidae family.

为了制备针对小反刍兽疫病毒(Peste des peties ruminants virus,PPRV)F基因的核酸疫苗,并评价其对小鼠的免疫效果,试验采用DNA重组技术将F基因、优化密码子后的F基因以及连接乙型脑炎病毒信号肽(JEVSP)的优化密码子后的F基因3个目的片段分别克隆到真核表达载体pcD?NA3.1(+),构建重组质粒pcDNA3.1(+)-F、pcDNA3.1(+)-F-opt和pcDNA3.1(+)-JEVSP-F-opt,大量培养重组质粒菌后,以碱裂解法将菌体悬浮裂解,经中空纤维切向流超滤和微滤浓缩处理,将制备的核酸疫苗以100μg/只的剂量给Balb/c小鼠肌肉注射免疫,共分为4组,pcDNA3.1(+)-F组、pcDNA3.1(+)-F-opt组、pcDNA3.1(+)-JEVSP-F-opt组和pcDNA3.1(+)空载体组,每隔14 d免疫1次,共免疫3次,每次免疫14 d后利用中和试验检测抗体水平.结果表明:成功制备核酸疫苗pcD?NA3.1(+)-F、pcDNA3.1(+)-F-opt和pcDNA3.1(+)-JEVSP-F-opt,免疫Balb/c小鼠14 d后pcD?NA3.1(+)空载体免疫组中和抗体阳性率为0,pcDNA3.1(+)-F组、pcDNA3.1(+)-F-opt组、pcD?NA3.1(+)-JEVSP-F-opt组抗体转阳性,且pcDNA3.1(+)-F-opt组、pcDNA3.1(+)-JEVSP-F-opt组抗体水平呈逐渐增加的趋势.说明成功制备小反刍兽疫核酸疫苗,并且能在小鼠上产生抗体.

In the present study, the complete sequence of the mitochondrial genome (mitogenome) of Papilio paris (Lepidoptera: Papilionidae) is described. The mitogenome (15,347 bp) of P. paris encodes 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and an adenine (A) + thymine (T)-rich region. Its gene complement and order is similar to that of other sequenced Lepidopterans. Phylogenetic analyses based on 13 PCGs using maximum-likelihood (ML) revealed that P. paris resides in the Papilionoidea family. This study provided the valuable evidence on phylogenetic relationship of the P. paris at the molecular level and essential resource for further research on this species.

为探明新疆某羊场疑似羊口疮和羊胸膜肺炎感染死亡羊只的病原,本试验在无菌条件下对送检肺脏病料进行细菌培养,并用支原体引物进行PCR扩增,后利用羔羊睾丸细胞对处理后的唇部痂皮病料进行毒株分离,分离到的毒株用羊口疮特异性引物进行PCR扩增和动物回归试验.结果显示,根据细菌培养和PCR扩增判断死亡羊只的传染性胸膜肺炎阳性,用羊睾丸细胞成功分离到的毒株经PCR验证显示羊口疮阳性,通过动物回归试验羊口疮攻毒组羊只出现口唇部脓疱痂皮等羊口疮典型病变,综合说明死亡羊只为传染性胸膜肺炎和羊口疮病原混合感染.本试验成功分离到一株羊口疮毒株且该毒株有较强的致病性,为今后进一步研制羊口疮疫苗打下了良好的基础.

为明确从新疆某绵羊养殖场送检的病死羊的致病菌,对病死羊的心脏、肝和肺组织采用细菌分离培养、形态观察、生化试验,纯化菌落进行种特异性和荚膜血清型PCR鉴定以及药敏试验、致病性试验和动物回归试验.为了解分离菌株的生物学特性及免疫原性,笔者以BALB/c小鼠为动物模型对分离株进行同源性和异源性的免疫攻毒保护试验.结果 显示,在BHI琼脂培养基上长出灰色、光滑小菌落,革兰氏染色为红色的小短杆菌;分离株能发酵葡萄糖等,符合多杀性巴氏杆菌(Pm)的生化特性;PCR扩增出大小为460bp的种特异性条带和1 045 bp的荚膜血清A型特异性条带,Blast检索结果与Pm的同源性为99.7％.结果 表明,分离菌为多杀性巴氏杆菌,属于荚膜血清A型.药敏试验显示,该菌株对头孢拉定、环丙沙星、替米考星等7种药物具有敏感性;该菌株能够引起实验小鼠和兔的发病死亡,根据羔羊的发病情况和临床表现,本菌为强致病性菌株;将分离株培养物研制成不同佐剂的灭活疫苗探究其免疫效果,氢氧化铝佐剂灭活疫苗可以抵抗同源菌的攻击,保护率能够达到100％;对异源血清型Pm免疫攻毒保护效果差,对D型、F型和未知型的保护率分别为20％、20％、0.本研究成功分离出1株绵羊A型多杀性巴氏杆菌,对羊巴氏杆菌病的流行监测和防控具有重要意义.

羊链球菌病是由链球菌引起的一种羊的急性、热性、败血性传染病.本病例根据其发病情况、临床症状和剖检变化做出初步诊断,然后依据血常规检查(WBC、Neu/％升高,Lym％下降)、病菌分离培养(镜检革兰染色阳性、呈长短链排列的球菌)、生化试验(过氧化氢酶试验阴性)及细菌PCR鉴定,最终确诊为绵羊链球菌病.通过及时的诊断与治疗,羊群的病情得到有效控制,合理化的药物选择和积极的防控措施是治愈本病的关键.

犊牛腹泻是常由多种病原微生物引起的一种急性传染病,是危害犊牛健康的最大疾病之一.为了降低规模养殖场腹泻犊牛的发病率和死亡率,本文简要分析规模化养殖场犊牛腹泻的致病原因,并提出了防治措施,以期为犊牛腹泻的防治提供参考.

[目的]为探明一起牛运输热的病原及生物学特性,[方法]研究采集病死牛心血、肺脏,对其进行细菌分离、生化试验和PCR鉴定,并对分离株进行毒力基因检测、致病性研究.[结果]结果显示,该分离菌为革兰氏阴性菌,呈球状或短杆状、两端钝圆、两极浓染.生化试验结果显示,分离菌能发酵葡萄糖、麦芽糖、阿拉伯糖、甘露醇、甘露糖、乳糖、木糖等碳水化合物,不发酵脲酶和吲哚,产生少量酸而不产气,符合溶血性曼氏杆菌的生化特性.PCR鉴定为荚膜血清A1型溶血性曼氏杆菌,并完成了毒力基因的检测.[结论]结果表明,引起该批牛运输热的病原为携带毒力基因的荚膜血清A1型溶血性曼氏杆菌,本研究结果为进一步研究溶血性曼氏杆菌的致病机制、防控措施等提供参考.

目的 探讨小反刍兽疫病毒(Peste des Petits Ruminants Virus,PPRV)N蛋白原核表达、条件优化及反应活性.方法 参照GenBank中PPRV (Nigeria/75/1)N基因序列,人工合成该基因,构建重组表达质粒pET-30a(+)-N,转化至BL21 (DE3)感受态细胞,IPTG诱导表达,进行SDS-PAGE电泳、Western-blot分析.在不同温度、时间、IPTG浓度条件下诱导表达N蛋白,确定最佳表达条件.结果 PPRV N蛋白(64.4 kD)成功表达,能被羊PPRV免疫血清所识别.N蛋白主要以包涵体存在,其最佳诱导条件:诱导温度28℃、IPTG终浓度2.0 mmol/L、诱导时间16 h.结论 原核表达的PPRVN蛋白具有良好的特异性和反应活性,为单克隆抗体的制备及快速诊断方法的建立提供了技术资料.

In the present study, the complete sequence of the mitochondrial genome (mitogenome) of Parasa tessellata (Lepidoptera: Limacodidae) is described. The mitogenome (15,308 bp) of P. tessellata encodes 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and an adenine (A) + thymine (T)-rich region. Its gene complement and order are similar to that of other sequenced lepidopterans. Phylogenetic analyses based on 13 PCGs using maximum likelihood (ML) and Bayesian inference (BI) revealed that P. tessellata resides in the Sphingidae family. This study provided the valuable evidence on phylogenetic relationship of the P. tessellata at the molecular level and essential resource for further research on this species.

The three croakers (Nibea coibor, Protonibea diacanthus and Argyrosomus amoyensis, Perciformes, Sciaenidae) are important commercial species inhabiting the Eastern Indian Ocean and Western Pacific. Molecular data employed in previous research on phylogenetic reconstruction have not been adequate and complete, and systematic and comprehensive phylogenetic relationships for these fish are unresolved. We sequenced the complete mitochondrial genomes of the three croakers using next-generation sequencing for the first time. We analyzed the composition and phylogenies between 19 species in the family Sciaenidae using the mitochondrial protein coding sequences of 204 species in the Series Eupercaria. We present the characterization of the complete mitochondrial genome sequences of the three croakers. Gene arrangement and distribution of the three croakers are canonically identical and consistent with other vertebrates. We found that the family Sciaenidae is an independent branch that is isolated from the order Perciformes and does not belong to any extant classification. Therefore, this family is expected to belong to a new classification at the order level and needs further analysis. The evolution of Sciaenidae has lagged far behind the Perciformes differentiation. This study presents a novel insight into the phylogenetics of the family Sciaenidae from the order Perciformes and facilitates additional studies on the evolution and phylogeny of Series Eupercaria.

The apple snails Pomacea canaliculata, Pomacea diffusa and Pomacea maculate (Gastropoda: Caenogastropoda: Ampullariidae) are invasive pests causing massive economic losses and ecological damage. We sequenced and characterized the complete mitochondrial genomes of these snails to conduct phylogenetic analyses based on comparisons with the mitochondrial protein coding sequences of 47 Caenogastropoda species. The gene arrangements, distribution and content were canonically identical and consistent with typical Mollusca except for the tRNA-Gln absent in P. diffusa. An identifiable control region (d-loop) was absent. Bayesian phylogenetic analysis indicated that all the Ampullariidae species clustered on the same branch. The genus Pomacea clustered together and then with the genus Marisa. The orders Architaenioglossa and Sorbeoconcha clustered together and then with the order Hypsogastropoda. Furthermore, the intergenic and interspecific taxonomic positions were defined. Unexpectedly, Ceraesignum maximum, Dendropoma gregarium, Eualetes tulipa and Thylacodes squamigerus, traditionally classified in order Hypsogastropoda, were isolated from the order Hypsogastropoda in the most external branch of the Bayesian inference tree. The divergence times of the Caenogastropoda indicated that their evolutionary process covered four geological epochs that included the Quaternary, Neogene, Paleogene and Cretaceous periods. This study will facilitate further investigation of species identification to aid in the implementation of effective management and control strategies of these invasive species.