共查询到19条相似文献，搜索用时 312 毫秒
1.
旨在探讨多房棘球蚴感染早期对小鼠肝脂质代谢的影响。选取60只清洁级BALB/c小鼠(6周龄，雄性)随机分为试验组和对照组，每组30只。试验组小鼠每只腹腔接种600个原头蚴，对照组小鼠腹腔注射等体积的PBS缓冲液。采用原位两步循环灌流及多步低速差速离心分离纯化肝实质细胞。利用高通量测序分析感染3月小鼠肝实质细胞差异表达的mRNA,并对其进行KEGG、GO分析和qRT-PCR验证。制备多房棘球蚴感染2、3和6月小鼠的肝组织切片，分别进行油红O染色，并通过qRT-PCR检测多房棘球蚴感染不同感染时期肝实质细胞脂质代谢相关基因的表达水平。结果表明，与未感染对照组相比，小鼠感染多房棘球蚴后3、6月肝脂肪沉积明显增多；感染2月的肝实质细胞脂质合成相关基因Scd1、Fasn和Srebf1,以及脂质氧化分解相关基因Acox1和Cpt1α均明显上调；感染3月的肝实质细胞Scd1、Fasn、Srebf1及Cd36的表达水平均明显升高，而Acox1、Cpt1α、Pparγ和Pparα的表达水平明显降低。上述结果表明，多房棘球蚴感染早期可能通过促进肝实质细胞的脂质合成，抑制脂质氧化分解，从而导致肝脂质沉积。 相似文献
2.
《畜牧兽医学报》2018,(11)
旨在解析miRNA在细粒棘球绦虫原头蚴中的表达谱特性,为进一步揭示miRNA在细粒棘球绦虫原头蚴发育中的作用及调控机制提供理论依据。从患病绵羊肝无菌采集细粒棘球绦虫原头蚴,提取总RNA,构建原头蚴miRNA文库,利用RNA sequencing技术进行测序分析。结果表明,本研究从细粒棘球绦虫原头蚴中分离得到109个已知的miRNA分子,同时得到189个新的miRNA发夹前体分子。靶基因的预测结果显示,已知miRNA共有132个靶基因,而新发现的miRNA共有3 152个靶基因。进一步的研究发现这些miRNA分子广泛参与虫体生长发育相关的关键信号通路,诸如wnt、cAMP、Hedgehog、NF-κB、Notch以及胆酸盐等信号通路,研究又进一步采用实时荧光定量PCR以及原位杂交技术对虫体经典wnt信号通路相关的miRNA分子以及其靶标基因进行了差异表达以及组织定位研究。结果提示,HG328413.1_36258和β-catenin、HG328414.1_36364*和dis以及HG328399.1_25846和axin可能存在潜在的调控关系。综上所述,本研究在细粒棘球绦虫原头蚴中发现了大量的新的miRNA分子,丰富了原头蚴miRNA的数据,为进一步解析miRNA在细粒棘球绦虫原头蚴发育中的作用奠定了基础。 相似文献
3.
为观察多房棘球蚴不同地理株在蒙古沙鼠肝组织的发育状况及其对肝组织损伤情况的差异，本试验将欧洲株和中国宁夏株多房棘球蚴分别腹腔注射蒙古沙鼠，饲养并观察2个月，直至腹部出现明显肿大，然后对试验组和空白对照组蒙古沙鼠进行安乐死，剖检分离肝脏然后进行肝组织切片染色观察，并将不同地理株试验组和空白对照组进行比对分析。结果显示：2株不同地理株多房棘球蚴均能造成肝组织窦间隙增宽、新生血管增多并表现出不同程度的肝细胞损伤，但欧洲株多房棘球蚴肝组织窦间隙增宽较宁夏株明显，而宁夏株肝组织新生血管较欧洲株多；中国宁夏株棘球蚴生发层中有大量的原头蚴，欧洲株未见清晰的原头蚴。结果表明腹腔注射多房棘球蚴后，蒙古沙鼠肝组织受到破坏，不同地理株棘球蚴在感染后一定时期内对肝组织损害程度有差异，且中国宁夏株和欧洲株在蒙古沙鼠肝脏内的发育速度和原头蚴增殖能力有所不同。 相似文献
4.
旨在研究羊传染性脓疱病毒（orf virus，orfv）感染山羊皮肤成纤维细胞（goat skin fibroblasts cell，GSF）对GSF细胞microRNA（miRNA）表达谱影响，探究miRNA在orfv感染过程中的作用及调控机制。分别提取感染和未感染orfv的GSF细胞总RNA，构建miRNA文库，利用高通量测序技术进行miRNA差异表达分析，对差异表达miRNA靶基因进行预测，并进行GO和KEGG分析，随机选取10个差异miRNA进行RT-qPCR验证。结果显示，orfv感染组和未感染GSF细胞组相比共有678个显著差异表达的miRNA（fold change≥1.5），其中，上调表达miRNA有509个，下调表达miRNA有169个，uniq_miRNA的Venn图分析显示，感染组和对照组共有的miRNA仅占8.21%；GO和KEGG分析显示，差异表达miRNA主要参与脂质代谢、受体及细胞因子信号转导等细胞生物学过程，RT-qPCR验证结果与高通量测序结果一致。本研究结果表明，orfv感染GSF细胞对其编码的miRNA有显著影响，获得大量GSF细胞编码的与orfv感染相关的差异miRNA，为进一步从宿主miRNA层面揭示orfv感染和致病机制提供了参考依据。 相似文献
5.
《蚕业科学》2020,(2)
二分浓核病毒(bidensovirus,BDV)是家蚕病毒病的主要病原之一。为了筛选和鉴定家蚕体内与BDV感染相关的差异表达microRNA(miRNA),对家蚕品种JS的3龄起蚕添食BDV,36 h后收集家蚕并提取总RNA,采用高通量测序技术分析家蚕感染BDV后应答的miRNA,从中筛选出24个显著差异表达的miRNA,其中14个为已知miRNA,10个为新发现的miRNA。进一步利用反转录PCR(RT-PCR)技术对上述24个miRNA进行验证,发现这些miRNA均在家蚕幼虫体内表达;利用实时荧光定量PCR(qPCR)技术检测家蚕感染BDV后这些miRNA的表达情况,发现其中9个miRNA表达上调,15个miRNA表达下调,其中表达上调倍数最高的为bmo-miR-1923,相对表达上调了8.87倍,表达下调倍数最高的为bmo-miR-13b-3p,相对表达下调了6.97倍。此外,根据测序结果进行生物信息学预测和分析差异表达miRNA的靶基因,并利用GO分析对靶基因进行功能分类,发现这些靶基因主要参与了细胞过程、催化、结合和代谢过程。研究结果将为阐明家蚕抗BDV的分子机制提供数据参考和技术支持。 相似文献
6.
藏鸡不同发育阶段腿部肌肉组织转录组及microRNA联合分析 总被引：1，自引：0，他引：1
旨在从转录组和miRNA角度探讨藏鸡肌肉发育的调控机制,了解藏鸡肌肉发育的特殊性。本研究对120和150日龄藏鸡的肌肉组织进行转录组和small RNA测序,筛选出两个日龄阶段差异表达的基因和miRNA,并利用qRT-PCR技术对测序结果进行验证。结果,共筛选出1 691个差异表达基因,其中上调基因330个,下调基因1 361个。差异表达miRNA共有22个,其中9个上调,13个下调。随机选择的5个基因和miRNAs的qPCR验证结果表明表达趋势与测序结果一致。GO富集分析显示,在富集前10的条目中,与免疫相关的条目占较大比例。KEGG分析结果显示,在19个显著富集通路中,与免疫相关的通路占较大比例,且83个miRNA的靶基因出现在显著富集通路上。转录组和small RNA测序数据联合分析表明,343个miRNA-mRNA对为负相关调控模式。本研究从多组学层面为进一步理解藏鸡的肌肉发育提供了理论基础。 相似文献
7.
《畜牧兽医学报》2019,(12)
旨在从转录组和miRNA角度探讨藏鸡肌肉发育的调控机制,了解藏鸡肌肉发育的特殊性。本研究对120和150日龄藏鸡的肌肉组织进行转录组和small RNA测序,筛选出两个日龄阶段差异表达的基因和miRNA,并利用qRT-PCR技术对测序结果进行验证。结果,共筛选出1 691个差异表达基因,其中上调基因330个,下调基因1 361个。差异表达miRNA共有22个,其中9个上调,13个下调。随机选择的5个基因和miRNAs的qPCR验证结果表明表达趋势与测序结果一致。GO富集分析显示,在富集前10的条目中,与免疫相关的条目占较大比例。KEGG分析结果显示,在19个显著富集通路中,与免疫相关的通路占较大比例,且83个miRNA的靶基因出现在显著富集通路上。转录组和small RNA测序数据联合分析表明,343个miRNA-mRNA对为负相关调控模式。本研究从多组学层面为进一步理解藏鸡的肌肉发育提供了理论基础。 相似文献
8.
《畜牧兽医学报》2017,(5)
本研究旨在分析山羊副流感病毒3型(CPIV3)感染MDBK细胞和正常MDBK细胞差异表达的miRNA,以探索miRNA在CPIV3感染过程中的作用及其调控机制。将CPIV3感染MDBK细胞,利用高通量测序技术检测分析MDBK细胞中miRNA表达谱,并筛选差异表达的miRNA。对靶基因进行GO注释和KEGG富集。结果显示,有37个miRNA显著差异表达(P0.001),其中18个miRNA在病毒感染中上调,19个miRNA在病毒感染中下调。经RT-qPCR方法验证5个差异表达的候选miRNA,结果与高通量测序分析有很好的一致性。进一步GO功能注释表明:差异表达的miRNA靶基因生物学功能相对集中在免疫调节、细胞生物调控及细胞新陈代谢等生物过程。这些靶基因参与多种细胞的信号通路,包括细胞黏附分子、B细胞受体信号通路、MAPK信号通路、代谢通路、黏合斑、钙离子信号通路和趋化因子信号通路等。本研究为进一步探讨CPIV3致病机制奠定了基础。 相似文献
9.
MicroRNA（miRNA）是一类约22个核苷酸组成的单链非编码RNA,在分化、发育、肿瘤形成等方面起着重要作用。本研究对弓形虫RH株感染小鼠脾细胞miRNA的表达进行了特异性基因芯片检测分析,并应用荧光定量RT-PCR方法进行验证。结果表明,感染鼠脾细胞中与免疫应答及细胞增殖和肿瘤发生相关的三大类miRNA中,有39种表达下调,同时有36种表达上调。上述结果揭示,弓形虫感染机体后伴随着靶细胞功能性miRNA表达谱的显著改变,这为进一步研究弓形虫感染致病的分子机制开辟了新的方向。 相似文献
10.
MicroRNA(miRNA)是在真核生物中发现的一类内源性的具有调控功能的非编码小RNA,在鸡的许多生物学功能中发挥重要作用,但在鸡的应激性免疫抑制的分子调控机制中尚不完全清楚。本试验通过在日粮中添加外源皮质酮激素,构建雏鸡应激性免疫抑制模型,高通量Solexa测序技术建立雏鸡法氏囊差异miRNA表达谱,共鉴定出77个差异表达的miRNAs,34个上调,43个下调,生物学分析预测差异表达miRNA的靶基因有282个,对这些预测的靶基因进行GO和KEGG通路富集分析,结果显示这些通路主要富集到心肌收缩、精氨酸和脯氨酸代谢、RNA聚合酶、肾素-血管紧张素系统。试验结果为揭示家禽应激性免疫抑制转录后分子调控机制提供了依据。 相似文献
11.
Vervaeke M van der Giessen J Brochier B Losson B Jordaens K Verhagen R Coulander Cde L Teunis P 《Preventive veterinary medicine》2006,76(3-4):137-150
The occurrence of the fox tapeworm Echinococcus multilocularis in Red foxes was studied in Belgium and a neighbouring region in The Netherlands. A total number of 1202 foxes were analysed (1018 in Belgium and 184 in The Netherlands) of which 179 were infected with E. multilocularis (164 in Belgium and 15 in The Netherlands). Further, the spatial distribution of infection among sampled foxes was analysed with an ellipsoidal gradient, demonstrating a decreasing prevalence in northwestern direction. Using this gradient, we showed that the spatial patterns of infection in Belgium and the neighbouring region in The Netherlands correspond, indicating a continuous distribution of E. multilocularis across the nation borders. Part of the Belgian data allowed investigating temporal changes in the spatial distribution of E. multilocularis. This revealed a northwestern spread of E. multilocularis. 相似文献
12.
Liver lesions were found in 6/6 pigs 7 months after oral inoculation with 5000 or 35,000 Echinococcus multilocularis eggs. However, lesion morphology differed considerably among the animals. The largest lesions (3–8 mm in diameter) were found in a single pig and smaller lesions (1.5–3 mm) in three pigs. These lesions were clearly circumscribed and had pronounced central necroses and dystrophic calcifications. In contrast, most of the smallest (usually <1.5 mm in diameter) found in two other pigs, had small compact fibrotic areas and blurred borders with obvious fibrous infiltrations into the interlobular tissues. E. multilocularis specific DNA was detected by PCR in all lesion types, but metacestode viability, as assessed by in vivo intraperitoneal inoculations in jirds, could not be demonstrated. Within 1 month post inoculation, all pigs developed specific IgG antibody responses against a battery of different antigens (metacestode, cyst fluid, and protoscoleces-derived native E. multilocularis and E. granulosus antigens, affinity purified Em2G11 antigen, antigen B, recombinant Em II/3–10 antigen). Two different reaction patterns were recorded. In the two pigs with the small lesions, pronounced reactions against all crude antigens with peaks 3–5 months p.i. and clearly elevated levels until the end of the experiment were noted. In all other pigs, antibody reactions remained low in all cases. In conclusion, we demonstrated two types of E. multilocularis metacestode development in pigs with distinct immunological response patterns. 相似文献
13.
为探究撒坝猪源大肠杆菌（E.coli）高致病性毒力岛（HPI）诱导小鼠病理损伤的超微结构特征,本研究将实验室保存的E.coli HPI阳性株（HPI+）和E.coli HPI基因缺失株（ΔHPI）进行复苏和培养,分别用E.coli HPI+、E.coli ΔHPI菌株以腹腔接种的方式感染昆明小鼠,检测菌株的半数致死量（50% lethal dose,LD50）,通过HE染色和透射电镜观察并分析菌株对小鼠病理损伤的超微结构特征,利用免疫组织化学标记白细胞介素-1β（interleukin-1β,IL-1β）阳性细胞在被感染小鼠肝脏和肾脏组织中的分布,以反映E.coli HPI+及E.coli ΔHPI菌株所引起炎症水平的差异。结果显示,E.coli HPI+和E.coli ΔHPI菌株的半数致死量分别为1×107.39和1×108.62 CFU/mL;HE染色显示,E.coli感染小鼠后,可见肝脏细胞肿胀、变性,肝窦淤血,肾脏间质淤血,肾小管上皮细胞变性脱落等病理变化;超微结构变化显示,肝脏细胞的完整形态消失,胞核呈不规则形态,线粒体畸形,粗面内质网上核糖体脱落,滑面内质网增生;多数肾小管上皮细胞出现胞核固缩,部分细胞核核仁边移、体积增大,足突融合,系膜细胞间隙变宽。此外,E.coli HPI+感染组小鼠于肝脏、肾脏的水肿现象较E.coli ΔHPI菌株感染的小鼠更为明显。免疫组化结果显示,大肠杆菌感染小鼠后,IL-1β蛋白主要表达于肝细胞、中央静脉周围、肾间质细胞和肾小管上皮细胞,且E.coli HPI+感染组的IL-1β表达量高于E.coliΔHPI感染组。综上所述,撒坝猪源E.coli HPI能够调控E.coli对小鼠的致病性,HPI的调控作用可使E.coli对小鼠肝脏、肾脏造成的病变及超微结构变化更明显,并且能够增加小鼠的炎症反应。 相似文献
14.
本研究旨在探讨猪m6A甲基化酶WTAP表达水平与大肠杆菌（E. coli）感染抗性的关系。选取35日龄苏太断奶仔猪（Sus scrofa）大肠杆菌抗性型和敏感型个体各4头,采集十二指肠和空肠组织,利用RT-qPCR检测WTAP在E. coli抗性型和敏感型个体十二指肠、空肠的表达差异,并分别利用产肠毒素大肠杆菌（F18ab、F18ac）刺激和内毒素（LPS）诱导猪小肠上皮细胞（IPEC-J2）,检测WTAP基因的表达变化。同时构建WTAP基因干扰载体并转染IPEC-J2细胞,通过菌毛定量、菌落计数以及间接免疫荧光试验检测该基因沉默对大肠杆菌黏附能力的影响。结果显示：在十二指肠和空肠组织中,WTAP基因在E. coli抗性型个体中的表达量显著高于敏感型个体（P<0.01）;并且在F18ab和F18ac刺激后表达量显著下降,与LPS诱导6 h后结果相一致（P<0.01）。沉默WTAP基因后,大肠杆菌黏附能力极显著上升（P<0.01）。本研究在细胞和个体水平上验证发现,m6A甲基转移酶WTAP的高表达可能有助于仔猪抗大肠杆菌感染,为进一步揭示仔猪抗大肠杆菌感染的RNA甲基化调控机制奠定基础。 相似文献
15.
Gram-positive cocci were isolated from half a month diseased lambs with a fatal infection in a sheep farm in Minhe county, Qinghai province.The clinical symptoms observed were drooping, reduced feed intake, emaciated and fever.The infected lambs died several hours after the disease been observed.Necropsy of two dying lambs revealed that for one lamb, the liver was enlarged and capsule of the liver dropped, and other organs had no obvious changes;For the other one, the organs had no obvious changes.After gram staining, gram-positive cocci were observed from all the samples.Partial Tuf gene sequences of these isolates were amplified and sequenced for species identification, BLASTn conparison result revealed that they had the highest sequence similarity with that of Enterococcus faecium (E.faecium) AUS0085 (99.8%), we thus primarily confirmed that these strains belonged to E.faecium.Antimicrobial susceptibility test indicated that these strains could be divided into two classes.Class 1 was resistant to β-lactams, aminoglycosides, macrolides, tetracyclines, quinolones, rifamycins, lincomycin and clindamycin;Susceptible to chloromycetin and vancomycin.Class 2 was resistant to β-lactams, aminoglycosides, tetracyclines, quinolones, rifamycins;Intermediate resistance to erythromycin and roxithromycin;But susceptible to clarithromycin, azithromycin, clindamycin, lincomycin, chloromycetin and vancomycin.Multilocus sequence typing (MLST) based on seven housekeeping genes for E.faecium revealed that E.faecium lby1 strain in class 1 and E.faecium lbg3 strain in class 2 were defined as a new sequence type ST989.Phylogenetic tree analysis revealed the representative strain of these isolates, E.faecium lbg3, was clustered together with E.faecium ST468.The pathogenicity test revealed that E.faecium lby1 and lbg3 strains both had pathogenicity. 相似文献
16.
试验从青海民和县某羊场致病性感染半月龄羔羊中分离出革兰氏阳性菌,感染羔羊临床症状有精神萎靡,采食减少,发病1~2 h后死亡。两只濒死羔羊剖检结果发现,其中一只出现肝脏肿大及肝包膜脱落,其余脏器无明显病理变化;另一只各脏器均未发现病理变化。分离病料,革兰氏染色发现,从病料中分离到的细菌均为革兰氏阳性球菌。对分离菌株进行Tuf基因PCR扩增,BLASTn比对结果显示与屎肠球菌AUS0085核苷酸序列相似性为99.8%,据此初步确定这些菌株为屎肠球菌。药敏试验结果表明,这些菌株可分为两类,第1类对β-内酰胺类、氨基糖苷类、大环内酯类、四环素类、喹诺酮类、利福霉素类抗生素及林可霉素和克林霉素耐药,对氯霉素和万古霉素敏感;第2类对β-内酰胺类、氨基糖苷类、四环素类、喹诺酮类、利福霉素类抗生素耐药,对红霉素和罗红霉素中度耐药,但对克拉霉素、阿奇霉素、克林霉素、林可霉素、氯霉素和万古霉素敏感。对第1类耐药性代表菌株lby1和第2类耐药性代表菌株lbg3进行多位点序列分析(MLST),结果发现这些菌株为一个屎肠球菌的新序列型,为ST989。系统进化树结果显示屎肠球菌lbg3与屎肠球菌ST468亲缘关系最近,致病力试验结果表明屎肠球菌lby1和lbg3均具有致病性。 相似文献
17.
Liver glycogen levels of broilers infected with Eimeria acervulina, E. brunetti, or E. tenella fell during the acute phase of the infection with the maximum effect at 5–6 days post-inoculation (DPI). During the early recovery phase (6–8 DPI), liver glycogen levels in the E. acervulina-infected birds increased to levels up to 3 times greater than those found in uninoculated control birds. A lesser increase was occasionally seen in E. tenella-infected birds. Pair feeding studies showed that the decrease in liver glycogen was not related to the amount of feed consumed. The magnitude of the glycogen overshoot at 7 DPI was not related to the depression of weight gain at 5 and 6 DPI. When feed was withheld from birds, liver glycogen levels of uninoculated control birds fell rapidly within 1 h after feed withdrawal. In birds infected with E. acervulina, liver glycogen levels remained high even after 3 h starvation. Injection of glucagon indicated that glycogen could be mobilized in both infected and uninfected birds. 相似文献
18.
An avian pathogenic Escherichia coli strain M1000 showed a clear zone of erythrocyte lysis on sheep blood agar plates. The hemolytic activity was not detected in the culture supernatant nor was any DNA sequence homologous to the E. coli -hemolysin gene detected in the chromosome or plasmid DNA of the strain, indicating that the observed hemolysis was different from -type. To identify the genetic determinant responsible for the hemolysis, we performed random Tn5 insertional mutagenesis and obtained one mutant, named M5005, that totally lacked the hemolytic activity. Cloning and nucleotide sequencing of the region flanking the transposon insertion site in the M5005 chromosome revealed that the transposon was inserted within an open reading frame of the cyclic AMP receptor protein (CRP) gene, which is involved in one of the global regulatory networks of gene expression in E. coli. Nucleotide sequence analysis of the intact crp gene of the strain M1000 showed that the CRP protein of M1000 is 99% identical to that of K-12. Introduction of the intact crp gene on a low copy plasmid into the mutant M5005 restored the hemolytic phenotype, confirming that the mutation site in M5005 was in the crp gene. CRP plays a central role in catabolite repression, the phenomenon by which the synthesis of many enzymes required to metabolize various sugars is repressed in the presence of glucose. When the hemolytic activity of E. coli M1000 grown in the presence of glucose was examined, the hemolysis was totally impaired. These results indicate that the avian pathogenic E. coli strain M1000 produces a hemolysin the expression of which is dependent on crp gene function. 相似文献
19.
为建立牛源大肠杆菌-秀丽隐杆线虫致病模型，本研究从广西南宁、桂林、柳州等地区收集的牛病料中分离出13株大肠杆菌，继而对这些大肠杆菌进行血清学鉴定及小鼠、秀丽隐杆线虫的致病性试验。采用玻片凝集法鉴定大肠杆菌的O血清型，并将13株大肠杆菌培养液以3.0×109 CFU/mL、0.2 mL/10 g体重给小鼠腹腔注射，同时分别喂食N2野生型秀丽隐杆线虫。结果显示，大肠杆菌的O血清型为O127、O126和O44，其中O126为优势血清型（4/13）。致病性结果显示，有11株大肠杆菌能使小鼠致死（致死率为40%~100%），2株大肠杆菌对小鼠没有致死性（致死率为0）。对小鼠有强致病性的大肠杆菌，对线虫的致死率也较高，死亡率在第3~6天最为显著，半数致死时间为3~4.5 d，最长存活时间为9 d；对小鼠不致死的两株大肠杆菌对线虫的致死率也较低，线虫死亡率下降趋势缓慢，半数致死时间为5~6 d，最长存活时间为10~11 d。肠道细菌计数结果显示，大肠杆菌在线虫体内的数量与时间呈线性关系，大肠杆菌不断破坏线虫的免疫系统，从而导致了线虫的死亡。本研究结果表明，大肠杆菌对线虫和小鼠的致病力试验结果一致，说明成功建立了牛源大肠杆菌-秀丽隐杆线虫致病模型，为牛病防治与临床用药提供了依据。 相似文献