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《中国兽医学报》2015,(8):1239-1243
为建立简便、高效、稳定的绵羊气管上皮细胞的体外培养及鉴定的方法,并在所培养的细胞上研究绵羊肺炎支原体侵染模型,本研究利用链蛋白酶冷消化法对绵羊气管上皮细胞进行体外分离,通过差速贴壁法纯化气管上皮细胞并进行传代后用液氮保存,细胞复苏后通过测定生长曲线、上皮细胞骨架蛋白角蛋白8和18以及对染色体与微生物的检测等对所培养细胞进行鉴定,并将鉴定纯化的气管上皮细胞用绵羊肺炎支原体进行侵染。结果显示,成功培养出绵羊气管上皮细胞,纯化的细胞在显微镜下排列紧密,形态均一,呈鹅卵石铺路样。细胞生长曲线呈"S"型,分子生物学手段和增菌试验未检测到气管上皮细胞中含有微生物污染,通过细胞免疫组化检测到了细胞角蛋白8、18,染色体核型数目为2n=54。绵羊肺炎支原体能黏附于分离纯化的绵羊支气管上皮细胞,为进一步研究支原体侵染细胞的机制等奠定了基础。 相似文献
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肺炎支原体感染大鼠实验模型的建立 总被引：1，自引：0，他引：1
本试验旨在建立肺炎支原体感染的Wistar大鼠模型 ,为研究肺炎支原体的发病机制及药物治疗理论提供基础。采用滴鼻法对实验大鼠进行肺炎支原体感染 ,利用PCR法进行咽拭子检测 ,并以透射电镜和光学显微镜进行肺部病理组织学检查。结果发现 ,实验大鼠在感染肺炎支原体 1 0d时 ,咽拭子PCR检测结果为阳性 ;透射电镜观察到肺脏细胞膜破裂 ,线粒体变性 ,嵴断裂 ;光镜下可见到支气管及肺血管周围有明显的淋巴细胞浸润 ,形成斑片状间质性支气管肺炎。结果表明了Wistar大鼠对肺炎支原体较易感 ,并产生以间质性肺炎为主要特征的肺部和呼吸道感染 ,也说明本次造模试验成功 相似文献
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用绵羊肺炎支原体（ＭＯ）抗原免疫的ＢＡＬＢ／Ｃ小鼠，取血清抗体阳性免疫鼠的脾细胞与ＳＰ２／Ｏ细胞在聚乙二醇作用下进行融合，产生杂交瘤细胞、用间接ＥＬＩＳＡ法对杂交瘤细胞生长孔上清液进行检测，筛选阳性杂交瘤细胞株．以有限稀释法克隆１～２次，得到６株分泌抗绵羊肺炎支原体抗原的单克隆抗体（ＭｃＡｂ）杂交瘤细胞株．选择抗体分泌较高的１４Ａ６、７Ａ２７、Ｂ３８进行了较详细的研究，结果表明，其核内染色体数为９４～９６．抗体属性是：１株为ＩｇＧ２ａ、２株为ＩｇＧ１．用间接ＥＬＩＳＡ法对３株ＭｃＡｂ作符异性分析、该ＭｃＡｂ只特异地与绵羊肺炎支原体抗原发生反应，而不与猪肺炎支原体、山羊无乳症支原体抗原发生反应．将杂交瘤细胞注射ＢＡＬＢ／Ｃ小鼠诱生腹水．其抗体效价提高１００倍．杂交瘤细胞在液氮中保存１～２年后复苏仍能稳定地分泌抗绵羊肺炎支原体ＭｃＡｂ。 相似文献
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本试验目的是检测无针皮下接种疫苗所诱导的猪对临床疾病的抗性,并比较肌肉和皮下接种猪肺炎支原体疫苗时的血清学反应。材料与方法:在每一组试验中,猪都曾接种过1次或2次猪肺炎支原体疫苗,并且检测其血清学反应都为阴性。通过DAKO猪肺炎支原体EL ISA反应试剂盒来检测肌肉和皮下接种疫苗时的血清学反应,皮下接种疫苗的效果通过猪肺炎支原体异源强毒菌株的试验性呼吸激发来检测。试验结果:所有疫苗注射点的副反应都是最小的。猪通过有针和无针皮下接种疫苗与肌内接种疫苗相比,前者具有较高的猪肺炎支原体抗体滴度。无针皮下接种与不接种疫苗的猪相比,前者更具有低的肺病变分数和在支气管肺泡灌洗液中有较高的IgA和IgG滴度。单一皮下接种疫苗21 d后机体建立了免疫保护。无针皮下接种猪肺炎支原体疫苗较安全且有效,不需要剃毛或者其他的皮肤准备工作。结论:如果配制小剂量的疫苗(如0.2 m l)对猪进行无针皮下注射是可行的。 相似文献
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唐彩琰 《国外畜牧学(猪与禽)》2017,37(6)
尽管目前对猪肺炎支原体的免疫机制尚未完全清楚,但接种疫苗确实能有助于减少肺部损伤的大小和持续时间。疫苗中的抗原成分和佐剂成分对产生免疫应答至关重要。非产毒素性脂多糖似乎可促进体液和细胞介导的对猪肺炎支原体疫苗的反应。 相似文献
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为建立绵羊肺炎支原体和精氨酸支原体的双重PCR检测方法,本试验分别设计了绵羊肺炎支原体和精氨酸支原体的特异性引物,优化反应条件后对其特异性和敏感性进行评价,并对40份鼻拭子进行了检测。结果显示,该方法能同时扩增出绵羊肺炎支原体545 bp和精氨酸支原体806 bp的特异性片段,而对其他病原的DNA扩增均为阴性。该双重PCR方法对绵羊肺炎支原体和精氨酸支原体的最低检测限分别为100和10 pg/μL。40份鼻拭子检测结果显示,双重PCR检测方法与分离培养法符合率高达92.5%,均能鉴定出绵羊肺炎支原体和精氨酸支原体。结果表明,本研究建立的双重PCR方法可用于绵羊肺炎支原体和精氨酸支原体的临床快速诊断。 相似文献
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芩百清肺浓缩丸对支原体肺炎模型鼠肺组织影响的研究 总被引：1，自引：0，他引：1
采用滴鼻法建立大鼠肺炎支原体感染模型,并给予芩百清肺浓缩丸以观察其对鼠肺部组织的影响.光镜观察结果显示,感染鼠肺部病理变化为支气管及肺血管周围有明显的淋巴细胞浸润,形成斑片状间质性支气管肺炎.病变程度以模型组最重,芩百高剂量组与正常对照组相近;电镜扫描显示,模型组大鼠气管上皮表面被粘液和纤维蛋白等覆盖,纤毛粘连、倒伏,部分脱落,表现为明显的支气管慢性炎症,粘膜水肿,细支气管变厚而管腔变窄.严迪组和芩百高、中剂量组细胞界限较清晰,粘膜处于上皮修复和纤毛再生状态;咽拭子PCR检测结果为模型组阳性,芩百组和严迪组为阴性.以上结果表明,芩百清肺浓缩丸能有效地减轻支原体对肺部组织的炎性病变,促进气管上皮细胞的恢复. 相似文献
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为了确定宁夏某羊场发生一起以体温升高、咳嗽、呼吸困难为临床特征且具有传染性的疾病的病原，采集濒死病羊的病理组织，应用细菌分离培养、分子生物学鉴定、病理组织学观察及免疫组织化学分析等方法对采集的病料进行病原检测和诊断。结果显示，细菌分离培养获得3株支原体分离株，分子进化分析发现其均属于绵羊肺炎支原体，与国际标准株Y-98的序列一致性为100%。病理组织学观察显示，肺泡上皮细胞坏死、增生，支气管管腔内和肺泡腔内大量炎性细胞浸润，支气管和气管黏膜上皮细胞坏死、脱落、固有层水肿、炎性细胞浸润。对不同脏器的免疫组织化学分析显示，绵羊肺炎支原体主要集中于肺脏和气管内，且肺脏的阳性组织百分比最高，其次是支气管和气管。结合疫病的流行病学调查，该羊场所发疾病为运输应激后感染绵羊肺炎支原体导致，该病原侵染并定植于肺脏、气管及支气管上皮细胞，引起羊严重的呼吸道病症。 相似文献
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抗绵羊肺炎支原体单克隆抗体杂交瘤细胞株的建立 总被引：1，自引：0，他引：1
用绵羊肺炎支原体抗原免疫的ＢＡＬＢ／Ｃ小鼠，取血清抗体阳性免疫鼠的脾细胞与ＳＰ２／Ｏ细胞在聚乙二醇作用下进行融合，产生杂交瘤细胞、用间接ＥＬＩＳＡ法对杂交瘤一长孔上清液进行检测，筛阳性交瘤细胞株。 相似文献
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《Journal of Equine Veterinary Science》2014,34(11-12):1338-1341
Tracheal leiomyoma is a rare neoplasm of the respiratory tract in animals. This primary benign tumor arises from smooth muscle cells of the trachea. We report a 13-year-old female horse with clinical signs of severe airways obstruction. It had dyspnea, exercise intolerance, chronic progressive respiratory noises, and cyanosis in mucosa membranes. Radiography revealed a foreign body obliterating the luminal trachea. Endoscopic biopsy showed a 3 × 3-cm ovoid mass attached to the dorsal aspect of the trachea. Hematoxylin and eosin and Masson's trichrome stains showed dense spindle cells forming irregular bundles disposed in short interlacing fascicles. Nuclei were elongated with blunt ends, eosinophilic cytoplasm with indistinguishable cell borders, and low mitotic activity. This study is considered the first reported case of an obstructive intratracheal leiomyoma in the horse. 相似文献
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本试验旨在研究紫菀乙醇提物对豚鼠离体气管平滑肌收缩功能的影响。采用离体气管恒温灌流的方法,将豚鼠气管制成气管螺旋条,通过 BL-420F 生物机能实验系统测定其张力的变化,观察紫菀乙醇提物对离体气管平滑肌静息状态下的舒张作用,以及在氯化乙酰胆碱(Ach)、磷酸组胺(His)、CaCl2条件下和无钙下Ach诱导细胞内钙释放和外钙内流所致两种收缩条件下对离体气管平滑肌张力变化的影响。试验结果显示,低浓度紫菀乙醇提取物(0.002~0.008 g/mL)对静息状态下豚鼠离体气管平滑肌具有一定的收缩作用,高浓度(0.008~0.196 g/mL)时具有舒张作用;低、中、高3个剂量组均可抑制Ach、His和CaCl2引起的气管平滑肌收缩强度,使各致痉剂的量效曲线非平行右移并降低最大效应。紫菀乙醇提取物对His的抑制作用强度最大,其次为Ach,最后是CaCl2,且均呈剂量依赖性。以上结果表明紫菀乙醇提物具有舒张气管平滑肌的作用,其机制可能与抑制豚鼠气管平滑肌M受体、H1受体和阻断Ca2+通道从而抑制细胞Ca2+内流有关。 相似文献
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PENG Wen-jing XIN Rui-hua LIU Yan LUO Yong-jiang WANG Gui-bo LUO Chao-ying XIE Jia-sheng LI Jin-yu ZHENG Ji-fang 《中国畜牧兽医》2016,43(6):1572-1578
The aim of the experiment was to study the effect of alcohol extract of Aster tataricus L.f.on the contraction of isolated guinea pig tracheal smooth muscle.The guinea pig tracheal smooth muscle was prepared by isolated tracheal thermostatic perfusion method.The effects of Aster tataricus L.f.on tracheal smooth muscle under the resting state and contraction stimulated by acetylcholine (Ach),histamine (His),CaCl2,Ca2+ release and influx in cells without calcium were measured by BL-420F biological function experiment systems.The results showed that Aster tataricus L.f.could induce the contraction of isolated trachea at resting state at a low concentrations of 0.002 to 0.008 g/mL and exerted a relaxation on the isolated trachea when at the higher concentrations of 0.008 to 0.196 g/mL.All of the three concentrations of Aster tataricus L.f.could inhibit the tension stimulated by Ach,His and CaCl2 which were antispasmodic agents,led to the non-parallelled rightward moving of cumulative concentration-response curve and the decline of maximal responses.The suppressive effect on His was the strongest,followed by Ach and CaCl2,and the suppression could counteract the contraction induced by extracellular Ca2+influx significantly,and they were all expressed concentration-dependent manner.These results indicated the alcohol extract of Aster tataricus L.f.could relax tracheal smooth muscle by acting on M-receptor and histamine receptor and blocking Ca2+ passage,thus inhibiting extracellular Ca2+influx. 相似文献
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以butylidenephthalide（BP）为关键词,检索SciFinder文摘索引数据库与中国知网（CNKI）中有关当归Angelica sineisis（Oliv.）Diels的研究文献,对其含有的同系化合物丁烯苯酞类n-butylidenephthalide（BP）的研究进展加以综述,并对相关的参考文献进行追踪检索,归纳了BP的提取方法、药理作用与含量测定方法。BP主要存在于当归的挥发油中,通过液相制备和分离,测定的主要方法为HPLC-MS法,主要药理作用有抑制血小板聚集、抑制平滑肌细胞增生、抗心绞痛、松弛子宫与气管平滑肌、抗肿瘤生长、抗动脉粥样硬化等。 相似文献
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An ovarian mucinous cystadenoma was found in a 5-year-old female cynomolgus monkey (Macaca fascicularis). The tumor was composed of various sizes of multilocular cystic glands lined by a single layer of mucin-filled epithelium. Each of these cystic glands was surrounded by a large amount of solid fibrous stroma resembling smooth muscle. The ovarian surface epithelium showed partial invagination into the ovarian cortex, and a transition was observed between the surface epithelium and the mucinous cyst-forming epithelium. Immunohistochemically, the stromal cells were positive for alpha-smooth muscle actin and proliferating cell nuclear antigen. Ultrastructurally, the glandular epithelium had numerous mucinous secretory granules and microvilli. The stromal cells had numerous parallel microfibrils with focal density. It is rare to encounter evidence of a transition from the surface epithelium to the mucinous tumor epithelium and to show stromal smooth muscle proliferation in a mucinous cystadenoma. 相似文献
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鸡传染性支气管炎病理形态学及发病机理的研究 总被引：5，自引：1，他引：4
１５０只１８日龄雏鸡随机分为两组，试验组９０只接种传染性支气管炎病毒（ＩＢＶ－Ｍ４１）后３５天内分３０批依次于不同时间扑杀，作组织病理学、超微结构及病毒抗原位检查，对照组６０只作相同的检查。结果表明，ＩＢＶ攻击的靶器官是气管，气管的病变表现为粘膜上皮细胞的损伤和脱、残留的在皮细胞增殖形成复层上皮、粘膜固有层及粘膜下层大量淋巴细胞浸润及粘膜逐渐恢复的相互连续的病理过程。肺脏初级、次级支气管也有类似的 相似文献
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Tissues were obtained at necropsy from the nasal vestibule, turbinates, nasopharynx, trachea, tracheobronchial bifurcation, and lung from each of 10 clinically healthy calves persistently infected (PI) with bovine viral diarrhea virus (BVDV) serotype 2a. Tissues from the nasal vestibule were obtained by biopsy from five additional PI calves. Formalin-fixed tissues were processed for immunohistochemistry to localize the distribution of BVDV throughout the respiratory tract. Antigen distribution and intensity were subjectively evaluated. Throughout the respiratory tract, mononuclear leukocytes, vascular smooth muscle, and endoneural and perineural cells had BVDV immunoreactivity (BVDV-IR). Multifocally, squamous and ciliated columnar epithelium throughout the respiratory tract contained weak to moderate BVDV antigen. Viral antigen was not seen in goblet cells. BVDV-IR in mixed tubuloalveolar glands of the nasal cavity was weak to strong in serous secretory cells and ductular epithelium. Chondrocytes of the concha often contained BVDV antigen diffusely. Nasal mucus-secreting and tracheobronchial glands multifocally contained weak viral signal. In all cases, alveolar macrophages had moderate to strong BVDV-IR, whereas BVDV-IR in alveolar epithelial cells was weak to moderate. BVDV was present in interalveolar leukocytes and mesenchymal cells. Results indicate that serous secretions of the nasal cavity, productive viral infection of epithelium, and infected leukocytes in respiratory secretions are likely major sources of infectious BVDV from PI calves. The presence of BVDV antigen in respiratory epithelium is, at least, indirect support for the notion that this virus predisposes PI cattle to secondary microbial infections. 相似文献