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1.
实验性鸡马立克氏病羽髓病变研究   总被引:1,自引:0,他引:1  
通过光镜1电镜等方法对鸡马立克氏病羽髓组织进行了形态学研究。结果显示,羽髓远端坏死,羽髓中有多量肿瘤细胞浸润,肿瘤细胞呈弥漫性或局部性浸润,电镜观察,肿瘤细胞核异型性明显,核膜凹陷,核内出现假核内包含物,在一些羽髓中,巨噬细胞吞噬肿瘤细胞较多见,而且观察到一些肿瘤细胞发生调亡,结果表明,鸡马立克氏病羽髓病变十分明显,羽髓的形态学变化可用于鸡马立克氏病的诊断。  相似文献   

2.
为探究二烯丙基二硫化物(DADS)对马立克氏病肿瘤细胞系(MDCC-MSB-1细胞)的增殖抑制作用及其信号通路的调控机制,以MDCC-MSB-1细胞作为研究对象,采用CCK-8法检测不同浓度DADS分别作用MDCC-MSB-1细胞24、48、72 h后对细胞增殖抑制的影响。用0、80、160、240μmol/L DADS预处理MDCC-MSB-1细胞24 h后,光学显微镜观察细胞形态变化;Hoechst 33342核酸染色法检测细胞形态变化;caspase-8活性试剂盒检测细胞中caspase-8蛋白酶活性;并用荧光定量PCR法检测Fas/FasL信号通路关键因子mRNA的转录水平。结果显示,40μmol/L~240μmol/L的DADS抑制MDCC-MSB-1细胞的增殖呈时间剂量依赖性;随着浓度的升高,细胞形态逐渐出现细胞核核膜消失,染色体降解为多个片段并且边缘化,形成凋亡小体;240μmol/L的DADS可诱导MDCC-MSB-1细胞caspase-8蛋白酶活性及Fas、FADD、caspase-8、Bid和caspase-3 mRNA的转录水平极显著升高(P0.01)。结果表明,DADS可诱导MDCC-MSB-1细胞的凋亡,并活化其Fas/FasL死亡受体凋亡信号通路,可为鸡马立克氏病的治疗提供参考。  相似文献   

3.
马立克氏病发生在育成期到成鸡期,本病能使鸡肝、神经、卵巢、腺胃、肾等脏器发生肿瘤病变。有些养鸡户鸡群接种马立克氏疫苗后仍有10—50%鸡群中发生马立克氏病。笔者针对什么时间发病,表现什么样的临床症状以及对病死鸡病理变化进行二年观察。  相似文献   

4.
2019年5月盐城市某蛋鸡养殖场发生疑似马立克氏病病例:主要的临床表现为严重消瘦,肢体麻痹;剖检可见单侧坐骨神经肿大,肝脏和脾脏上出现弥漫性肿瘤;羽囊浸液琼脂扩散实验中,受检羽髓与阳性血清孔之间有沉淀线;PCR扩增获得马立克氏病毒特异性目的条带。确诊该病例为马立克氏病病毒感染。  相似文献   

5.
鸡马立克氏病毒为疱疹病毒科B亚群病毒。病毒存在于鸡体被感染细胞的细胞核中(其对外界抵抗力不强)及病鸡羽毛囊上皮细胞中,其能在体外环境中生存较长时间,是导致发病的重要病原。  相似文献   

6.
本试验应用光镜、电镜、流式细胞术、实时定量PCR技术研究了雌激素诱导小鼠胸腺萎缩过程中胸腺指数、胸腺显微和超微结构、胸腺细胞凋亡率及部分细胞因子和凋亡相关蛋白基因表达的变化。结果注射雌激素后,小鼠胸腺指数显著下降,胸腺中凋亡细胞数量显著上升,细胞因子TGF-β1表达量显著升高,IL-6表达量略为升高,而IL-7表达量显著降低;凋亡相关蛋白Caspase-3、Caspase-9、FADD表达量显著上升,FasL表达量略有上升,而Bcl-2表达量显著降低。表明雌激素可能一方面通过调节胸腺细胞因子抑制胸腺上皮细胞的增殖,另一方面通过激活Caspase级联程序的启动和执行阶段及Fas/FasL凋亡信号通路促进胸腺细胞的凋亡,从而诱导小鼠胸腺萎缩。  相似文献   

7.
为确诊思南县某养殖专业合作社饲养的肉鸡发病死亡原因,采集病鸡羽髓、血样及病死鸡肝脏、脾脏进行细菌分离培养、相关病毒琼脂扩散试验和PCR检测。结果:肝脏组织中分离出沙门氏菌;羽髓琼脂扩散试验检测出马立克氏病抗原,血清琼脂扩散试验检测出马立克氏病抗体;肝脏、脾脏PCR检测出马立克氏病病毒核酸。结论:综合诊断为马立克氏病与沙门氏菌病混合感染。  相似文献   

8.
采用HE、Giemsa染色、透射电镜以及DNA琼脂糖凝胶电泳等研究了水泡性口炎病毒(VSV)诱导BHK-21细胞凋亡的过程。结果显示:VSV感染BHK-21细胞后,光镜下可见细胞圆缩,细胞器固缩、核仁消失、染色质凝聚和核碎裂、凋亡小体出现;电镜下观察到染色质聚集形成典型的新月形,胞浆中充满大量空泡,细胞核因染色质凝聚也发生了空泡化;1%的琼脂糖凝胶电泳出现180-200bp整倍数的DNA梯形条带。结果表明,VSV诱导BHK-21细胞凋亡是其致细胞病变的主要表现形式之一。  相似文献   

9.
运用透射电镜术、原位末端标记技术(TUNEL)和流式细胞术对11只自然感染兔脑炎原虫獭兔的肾脏进行了细胞凋亡研究,5只健康长耳白兔用作对照。结果表明.凋亡细胞多见于远曲小管和集合管的上皮细胞。用透射电镜观察,初期的凋亡细胞,其细胞核的染色质高度凝集,边缘化,粗面内质网增多、扩张呈泡状;后期的凋亡细胞,其细胞质浓缩.细胞膜起泡,细胞核分裂成块状,最后形成膜包裹的凋亡小体。TUNEL法检测,病变轻微的细胞,胞核微缩,胞膜增厚;病变严重的细胞,其胞核体积明显缩小,胞膜皱缩,最后形成膜包裹的凋亡小体。流式细胞术检测,病兔可呈现明显的亚“G1”峰,凋亡细胞百分比与对照兔相比差异极显著(P=0.01),且病兔。肾细胞的周期也有所改变。结果表明,3种方法检测的病兔肾上皮细胞凋亡数均高于对照免。  相似文献   

10.
对江苏仪征某鸡场饲养的200日龄种鸡发生的内脏型马立克氏病(MD)进行了系统的病理学观察。肉眼所见以心脏、肾脏、卵巢出现大小不等的白色结节,肺脏、肝脏和脾脏出现弥漫性白色肉变病变,腺胃肿大如球状为主要特征。显微病理组织学变化主要为出现肉眼病变的各脏器的肿瘤细胞弥散性浸润,由大小不一、着色不均、有多级核分裂相的多形态淋巴样细胞构成,未见有肉眼病变的肌胃也出现肿瘤细胞浸润,少数器官如腺胃、卵巢还出现嗜异性白细胞浸润,其他脏器均无明显显微病变。组织器官的正常组织结构均遭到不同程度的破坏,尤其以肝脏、脾脏和肾脏为严重。结合实验室病原分离和MD抗体检测结果,确诊本病为内脏型马立克氏病自然感染,为临床上MD的诊断提供了一定的借鉴。  相似文献   

11.
The involvement of apoptosis was evaluated in lesions of endotoxemic piglets. A single injection with E. coli O111:B4 lipopolysaccharide (LPS) induced foci of coagulative necrosis in the liver and kidneys. No significant change was observed in these organs at 1.5 hr after LPS injection, but at 6 hr, epithelial cells with chromatin condensation or fragmentation and apoptotic bodies were visible. Foci of coagulative necrosis were formed within 24 hr after LPS inoculation. In and adjacent to the necrotic foci, dead hepatocytes with nuclear condensation or fragmentation were scattered. These dead cells were positively stained by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) methods. Electronmicroscopy revealed apoptotic cells with condensed or fragmented homogeneous nuclear chromatin, and necrotic cells with irregularly destroyed nuclei and cytoplasmic membranes. Apoptotic cell death were also observed in parietal cells of the stomach and lymphocytes in the lymphatic system. DNA ladders with approximately 200-bp multimers were observed in hepatic, renal and thymic samples prepared after 6 and 24 hr of LPS injection by agarose gel electrophoresis. These results suggest that apoptosis is involved in the pathology of swine endotoxemia.  相似文献   

12.
选择5月龄胎儿水牛的卵巢,采用石蜡切片、HE染色的光镜观察和DNA末端原位标记技术(TUNEL)进行了研究。结果表明:5月龄胎儿卵巢上分布着大量的凋亡细胞;在皮质部存在的大量腔前卵泡中,均匀分布有单个凋亡的卵泡细胞和卵母细胞,凋亡细胞的主要组织学变化是细胞缩小,核浓缩;DNA末端标记变化主要显示出TUNEL阳性。上述结果提示:水牛卵泡细胞和卵母细胞凋亡在胎儿时期已经存在,细胞凋亡是启动水牛腔前卵泡闭锁的潜在机理。  相似文献   

13.
为探讨圆弧青霉菌毒素—青霉酸对小鼠脾脏组织的毒性作用,采用TUNEL法和RT-PCR对染毒后脾脏细胞凋亡和Bcl-2、Bax及Fas/FasL等凋亡相关基因mRNA表达的影响进行研究。结果表明,随着青霉酸染毒剂量的增加,脾脏组织中细胞凋亡数量逐渐增多。染青霉酸低剂量组脾脏组织中Bcl-2、Fas和FasL的表达增加,Bax表达下降,高剂量组和中剂量组中Bax、Fas和FasL表达增加,Bcl-2表达下降,说明青霉酸可能通过促使Bax、Fas/FasL表达增加和Bcl-2表达降低来诱导小鼠脾脏细胞的凋亡。  相似文献   

14.
Dogs are the main domestic reservoirs of L. (L.) chagasi. Once in the vertebrate host, the parasite may cause visceral leishmaniasis, which can also be transmitted to humans. Infected symptomatic dogs show disorganization in the white pulp in spleen tissue and a reduction in T lymphocytes in peripheral blood. To investigate whether apoptosis is involved in white pulp disorganization and diminished T cell counts in peripheral blood, apoptotic T cells from the spleen and peripheral blood of dogs naturally infected with L. (L.) chagasi and presenting clinical manifestations were quantified and compared with healthy dogs. Thirteen symptomatic adult dogs infected by L. (L.) chagasi and six healthy dogs from a nonendemic area (controls) were included in the study. Samples from spleen and peripheral blood were used to quantify apoptosis in CD3 lymphocytes by flow cytometry using Anexin V and Multicaspase kits; the results were compared using the Mann Whitney test. The percentage of total T cells was lower in Leishmania infected dogs compared to healthy controls (P<0.05). Apoptosis levels in T cells from PBMC and spleen were higher in infected dogs than in controls (P<0.05). The least squares method test was used to determine the effect between the degree of structural organization of spleen white pulp and the percentage of apoptosis in the spleen. A significant effect on the level of white pulp morphological disorganization and percentage of apoptosis in spleen T cells was observed (F=20.45; P=0.0014). These data suggest that apoptosis is an important for the immunopathogenesis of canine visceral leishmaniasis.  相似文献   

15.
Apoptosis has been shown to be an important regulator of endometrium function. To clarify the regulation of apoptosis in the cat endometrium during the normal oestrus cycle, the expressions of the apoptosis‐related proteins (Bcl‐2 and Bax) and their correlation to the inhibitor of apoptosis protein Survivin were analysed using immunohistochemistry. The TUNEL technique (TdT‐mediated dUTP nick end labelling) was also used to detect DNA fragmentation characteristic of apoptotic cells. The results demonstrated that TUNEL labelling is not effective for the detection of apoptosis in cat endometrium. Survivin was expressed in the luminal and glandular epithelial cells of cat endometrium during all phases of the oestrus cycle. Survivin was localized in both the cytoplasm and nuclei of superficial and deep uterine gland cells during the luteal phase, while only cytoplasmic staining was observed during the follicular and anoestrus phases. Bax immunoreactivity in the cytoplasm of luminal and glandular epithelial cells as well as the smooth muscle cells of blood vessels was weak in the anoestrus phase. Compared with anoestrus, the intensity of Bax immunostaining was moderate in the follicular phase and increased dramatically in the luteal phase. Bcl‐2 immunostaining in the cytoplasm of luminal and glandular epithelial cells was moderate in the anoestrus phase. During the early follicular phase, cytoplasmic Bcl‐2 immunostaining was detected mostly in glandular epithelial cells. In the mid‐follicular phase, in glands, the amount of Bcl‐2 protein increased progressively from the superficial to the deep layer. In contrast, the expression of Bcl‐2 decreased in the secretory phase, being very low or absent in the mid‐ and late luteal phases. The overall results suggest that Survivin, Bax and Bcl‐2 proteins may cooperatively contribute to cell apoptosis and cell proliferation in the cat uterus during the oestrus cycle.  相似文献   

16.
The frequency and the distribution of apoptotic cells were investigated in formalin-fixed paraffin-embedded lymphoid tissues from healthy conventional pigs at four different ages (6 days, 2 months, 3.5 months and 5 months). Samples of tonsil, mesenteric lymph node, spleen, thymus and Peyer's patches were histologically processed and apoptosis evaluated with the TUNEL reaction and cleaved caspase-3 immunohistochemistry. In each technique, quantification of positive labelling was done for each particular lymphoid tissue area. The labelling pattern and distribution were similar for TUNEL and cleaved caspase-3. TUNEL stained mainly apoptotic bodies inside macrophages, but signal was also seen in free apoptotic bodies and in the nuclei of lymphocyte-like cells. The anti-cleaved caspase-3 antibody labelled mainly nuclei of lymphocyte-like cells. All tissues presented a similar distribution pattern of apoptosis, except for the 6-day-old group. In this group, a scattered distribution of positive cells was detected in tonsil, lymph node and spleen. In the tonsil and mesenteric lymph nodes from the older pigs, follicular areas presented higher amounts of positive cells than interfollicular areas. Moreover, the splenic white pulp showed more positive reaction than the red pulp, especially when they included germinal centres. In all groups, the follicular areas of ileal Peyer's patches presented more labelled cells than the dome and the lamina propria. In the thymus, the higher apoptotic rates were found in the cortex. In general, TUNEL yielded higher rates of positive cells than cleaved caspase-3 immunolabelling. A good correlation between the two techniques was found for thymus, tonsil and mesenteric lymph node, but not for Peyer's patches and spleen. This study describes a detailed histochemical characterization of apoptosis in pig lymphoid tissues using TUNEL and a cleaved caspase-3 immunolabelling at different ages. Moreover, our results indicate that TUNEL and cleaved caspase-3 techniques can be equivalent only when tissues have a high or low levels of apoptosis, since a considerable discrepancy was found in intermediate situations. Data from this study should be useful for future comparative studies under disease conditions.  相似文献   

17.
A total of 114 male chickens from three sire families of a commercial cross of White Leghorn chickens were infected with RB-1B Marek's disease (MD) virus at 21 days of age by exposing them to chickens previously inoculated with MD virus. The presence of virus in feather tips, feather pulp, and MD viral antibodies indicated all chickens became infected. The first virus-positive chickens were observed at 12 days postexposure (dpe). The frequency reached a maximum at 27 dpe and then decreased. At 80 dpe, when the experiment was terminated, no viral DNA was detected in the feather pulp of the surviving chickens (82%). Death from MD was first observed at 38 dpe and reached 18% by the end of the experiment, with spleen lesions being the major MD lesion. The viral genome titers in spleen extracts of chickens with MD lesions was negatively correlated with the time of death, and, similar to feather pulp, none of the surviving chickens was virus positive at the end of the experiment. Quantization of the viral genome titers in feather tip extracts at 27 and 38 dpe revealed a positive correlation with the presence of MD lesions, but only in the declining phase (38 dpe) and not at the peak (27 dpe) of the viral titer. Sire effects were significant, indicating the presence of genetic factors that affect viral proliferation. Again, significance was only observed at 38 dpe and not at 27 dpe. The results indicate that, in this commercial line, 1) all chickens were susceptible to infection via contact exposure, 2) all surviving chickens recovered from the viral infection, and 3) it is not sufficient to measure viral titers at a single time point when using viral titers as an endpoint for MD susceptibility.  相似文献   

18.
将雏鹅新型病毒性肠炎病毒(NGVEV)强毒CH株经尿囊腔途径人工感染10日龄鸭胚,应用透射电镜和超薄切片技术研究病毒在宿主细胞内的形态发生及各组织器官的超微结构变化。结果表明:感染后不同时间剖杀及死亡鸭胚的尿囊膜、肠、心、肝、脑和肌胃组织中,均观察到60~70nm的病毒粒子。病毒粒子主要通过与细胞膜融合而进入细胞质内,然后在细胞核内进行复制和装配。最后病毒粒子通过核膜和细胞膜破裂的方式被释放。病毒侵害的主要靶细胞包括鸭胚尿囊膜上皮细胞、肠上皮细胞、肠道平滑肌细胞、成纤维细胞、肝细胞、肌胃黏膜上皮细胞和心肌细胞等,表现为细胞核内外膜间隙严重扩张,细胞质整体结构严重空化。病毒侵害的主要靶细胞器包括粗面内质网和线粒体,表现为粗面内质网扩张呈囊状;尿囊膜上皮细胞的线粒体出现固缩和异常聚集变化,而其他组织细胞的线粒体均表现为肿胀和嵴断裂、消失。本试验还发现NGVEV可诱导宿主细胞发生严重的细胞凋亡现象,表现为细胞皱缩,胞核内染色质密度增高,核固缩成一个或数个团块凝聚在核膜周边,胞质浓缩深染并形成凋亡小体。  相似文献   

19.
It has been reported that apoptosis plays an essential role in controlling the physiological cell kinetics in the human and rodent endometrium but this type of death has never been studied in the porcine endometrium. The aim of this study was to investigate the apoptotic cell death in the porcine endometrium during the middle (Days 9-11) and late (Day 13) luteal phase, during the luteolysis (Day 15) and early follicular phase (Days 17-19) of the oestrous cycle. Apoptotic cells were identified by in situ DNA 3'-end labelling method. It was revealed that the greatest number of apoptotic cells in the luminal and glandular epithelium was found on Days 17-19 and on Day 15 of the oestrous cycle, respectively. In the stroma, the greatest number of these cells was found on Days 9-11. Our data have shown that in the porcine endometrium, both epithelial and stromal cells undergo apoptosis and that the number of apoptotic cells varies depending on the phase of the oestrous cycle.  相似文献   

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