体外棕榈酸诱导自噬调节奶牛淋巴细胞炎症通路

为探讨体外棕榈酸(Palmitic acid,PA)是否可以通过自噬调节奶牛淋巴细胞炎症信号通路的激活,分离健康奶牛淋巴细胞,采用3-MA(3-Methyladenin,细胞自噬抑制剂)和不同质量浓度PA作用于淋巴细胞,收集细胞及上清,利用qRT-PCR检测淋巴细胞LC3B、Beclin1、mTOR、UKL1、SQSTM1、IL-1β、IL-6和TNF-αmRNA表达情况;利用CCK-8和酶联免疫吸附法分别测定细胞活性和促炎细胞因子TNF-α、IL-6和IL-1β的释放量。CCK-8结果表明:PA显著抑制淋巴细胞活性(P0.01),添加自噬抑制剂3-MA后细胞活性显著增强(P0.01);qRT-PCR结果表明:与对照组(PA=0μg/mL)相比,PA处理组(不含3-MA)ULK1和LC3B mRNA表达极显著增加(P0.01),mTOR、Beclin1和SQSTM1 mRNA表达极显著降低(P0.01);IL-1β和IL-6 mRNA表达显著增强(P0.01),而TNF-α未表达;添加3-MA后,与1μg/mL PA处理组相比较,SQSTM1和Beclin1极显著增加,而LC3B mRNA表达极显著降低(P0.01);IL-1β和IL-6 mRNA表达显著下调(P0.01);PA增加促炎因子IL-6和IL-1β的释放(P0.01),显著抑制TNF-α释放(P0.01),而含有3-MA的PA混合处理组则显著降低(P0.01)。综上,PA可通过自噬调节奶牛淋巴细胞炎症信号通路的激活。     

Effects of supplementation with green tea by‐products on growth performance,meat quality,blood metabolites and immune cell proliferation in goats

Forty‐eight castrated male goats were used to determine the effects of feeding green tea by‐products (GTB) on growth performance, meat quality, blood metabolites and immune cell proliferation. Experimental treatments consisted of basal diets supplemented with four levels of GTB (0%, 0.5%, 1.0% or 2.0%). Four replicate pens were assigned to each treatment with three goats per replicate. Increasing dietary GTB tended to linearly increase the overall average weight gain and feed intake (p = 0.09). Water holding capacity, pH and sensory attributes of meat were not affected by GTB supplementation, while cooking loss was reduced both linearly and quadratically (p < 0.01). The redness (linear; p = 0.02, quadratic; p < 0.01) and yellowness (quadratic; p < 0.01) values of goat meat were improved by GTB supplementation. Increasing dietary GTB quadratically increased protein and decreased crude fat (p < 0.05), while linearly decreased cholesterol (p = 0.03) content of goat meat. The proportions of monounsaturated fatty acid, polyunsaturated fatty acid (PUFA) and n‐6 PUFA increased linearly (p < 0.01) and n‐3 PUFA increased quadratically (p < 0.05) as GTB increased in diets. Increasing dietary GTB linearly increased the PUFA/SFA (saturated fatty acid) and tended to linearly and quadratically increase (p ≤ 0.10) the n‐6/n‐3 ratio. The thiobarbituric acid‐reactive substances values of meat were lower in the 2.0% GTB‐supplemented group in all storage periods (p < 0.05). Dietary GTB linearly decreased plasma glucose and cholesterol (p < 0.01) and quadratically decreased urea nitrogen concentrations (p = 0.001). The growth of spleen cells incubated in concanavalin A and lipopolysaccharides medium increased significantly (p < 0.05) in response to GTB supplementation. Our results suggest that GTB may positively affect the growth performance, meat quality, blood metabolites and immune cell proliferation when supplemented as a feed additive in goat diet.     

稳定表达山羊SLAM受体的BHK-21细胞系的建立及应用

信号淋巴激活分子(SLAM)又称为CD150,是小反刍兽疫病毒(PPRV)和犬瘟热病毒(CDV)等麻疹病毒属病毒感染淋巴细胞的主要受体,在病毒侵入细胞中发挥重要作用。为建立稳定表达山羊SLAM(g SLAM)的真核细胞系,本研究将人工合成的g SLAM基因克隆至真核表达质粒p IRESpuro3中,并在该基因的3'端引入Flag标签序列作为分子标记,构建了重组质粒p IRES3-g SLAM。将该重组质粒转染BHK-21细胞,经嘌呤霉素加压筛选及采用表达绿色荧光蛋白(GFP)的重组PPRV病毒(r PPRV/GFP)感染鉴定后,筛选到稳定表达g SLAM基因的细胞系。r PPRV/GFP感染和western blot鉴定表明,无论是否有嘌呤霉素压力的存在,该细胞系在传代至第20代,仍能稳定表达g SLAM蛋白。由于g SLAM氨基酸序列与犬的同源性较高,以表达GFP重组CDV强毒株(r CDV/GFP)感染该细胞系,病毒可以感染且能形成明显的细胞病变,表明该细胞系可用于CDV强毒分离和致弱机制等相关研究。     

内毒素对大鼠小肠黏膜组织的影响及多价阳离子A的保护效应

旨在揭示大肠埃希菌内毒素(ET)对大鼠小肠黏膜的结构、绒毛长度、上皮内淋巴细胞(IEL)的数量和分布的影响,并探讨多价阳离子A(CA)对上述指标的保护效应。选用72只140g~150g SPF级SD大鼠随机分为3组,即对照组、ET组和CA保护组,经相应处理后分别在3、4、8、12h采集十二指肠、空肠组织作为检测样本,制备病理组织切片,HE染色并利用图像分析系统进行分析。ET组十二指肠和空肠绒毛长度在3、4、8、12h均显著低于对照组和CA保护组(P0.01),ET组十二指肠、空肠IEL数量在3、4、8、12h均显著低于对照组(P0.01);CA保护组十二指肠和空肠IEL数量在4、8、12h均显著高于ET组(P0.01)。结果显示,ET在不同程度上能够破坏小肠黏膜的正常组织结构,降低小肠绒毛长度,减少IEL的数量,从而影响小肠正常的吸收和免疫功能,而CA则能明显降低ET所导致的毒性作用,发挥其保护效应。     

Anatomical variation of arterial supply to the rabbit spleen

The rabbit, which is widely used as an experimental animal and is also popular as a companion animal, has a flat and elongated spleen with the longitudinal hilus running along its visceral surface. The spleen receives via the hilus an arterial supply that is essential for splenic nutrition and normal functioning. However, the distribution and variation of the arteries to the spleen have not been studied in detail. This study investigated anatomical variations of splenic arterial supply in 33 New Zealand White rabbits with a colored latex injection into arteries. We also examined whether the length of the spleen correlated with the number of the splenic branches of the splenic artery. The splenic artery always arose as the first independent branch of the celiac artery and ran along the splenic hilus to usually provide 6 (range, 3 to 10) splenic branches to the spleen. There was a moderate correlation (R=0.6) between the number of splenic branches and the longitudinal length of the spleen. The splenic branches often arose as a trunk or trunks in common with short gastric arteries. The number of common trunk(s) was usually 1 (range, 0 to 4). The data showed that the pattern and number of arterial branches to the spleen varied according to the individual animal, suggesting that such variations should be considered when performing experimental and veterinary surgical treatments in rabbits.     

雏鸡中枢免疫器官内B淋巴细胞变化规律

为了研究雏鸡在正常发育过程中免疫器官内B淋巴细胞的变化规律,以不同日龄的SPF鸡为研究对象,用流式细胞术检测鸡脾脏、胸腺及法氏囊内IgA+、IgG+及IgM+B淋巴细胞含量的变化情况。结果显示,在脾脏中,4日龄时B淋巴细胞含量比较多,7日龄时B淋巴细胞含量下降,随后,7至21日龄细胞含量逐渐升高;胸腺内则是在4日龄时IgM+B淋巴细胞含量相对较高,而IgA+、IgG+B淋巴细胞几乎没有;在法氏囊中,IgA+、IgG+及IgM+B淋巴细胞含量在4至14日龄时逐渐升高,并且IgA+和IgG+细胞含量在21日龄时基本趋于稳定,但是IgM+B细胞含量则在21日龄时有所下降。试验结果表明,雏鸡出壳后各免疫器官B淋巴细胞始终以IgM+细胞含量最多。     

Ikaros基因研究进展

Ikaros基因为淋巴细胞发育过程中不可缺少的转录因子,可调控许多免疫基因的表达,由于其重要的功能而在免疫学领域具有独特的地位。文章介绍了Ikaros基因的结构、功能、调控机制以及鱼类中近年来的研究进展,指出了Ikaros基因在淋巴系统发育过程中的重要作用。     

替米考星和甲硝唑对猪外周血淋巴细胞增殖的影响

采用细胞形态学观察、淋巴细胞标记和噻唑兰(MTT)法,探究替米考星和甲硝唑单独用药和联合用药对猪外周血淋巴细胞转化率、α-醋酸萘酯酶法阳性率和淋巴细胞增殖的影响。试验结果显示甲硝唑具有提高细胞免疫的功能。在体外试验时,终浓度为0.25~100μg/mL的甲硝唑可以促进淋巴细胞的增殖和转化;在体内试验时,浓度为2.5~10 mg/kg的甲硝唑可以增加成熟T淋巴细胞的数量。但是替米考星在体内和体外试验的结果却与阴性对照差异不显著,表明它不具有促进或抑制淋巴细胞增殖的能力,对机体的细胞免疫无影响。     

Cytopathological and histopathological diagnosis of canine splenic disorders

Objectives To determine (1) the common types of canine splenic disorders, and the breeds affected, that are diagnosed by cytopathological and histopathological examination in Sydney, Australia and (2) the accuracy of cytopathological examination compared with histopathological examination for the diagnosis of canine splenic disorders. Design 69 cytopathological and 51 histopathological diagnoses of canine splenic disorders presented to the Veterinary Pathology Diagnostic Services, The University of Sydney during 2006 and 2007 were tabulated and analysed; 17 cases examined both cytopathologically and histopathologically during 2001–07 were also analysed. Results The most common cytopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (29%), followed by no abnormalities detectable (28%), malignant neoplasms (20%), equivocal diagnoses (20%) and inflammatory disorders (3%). The most common breeds were Kelpie crosses and mixed breeds. The most common histopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (49%), followed by malignant neoplasms (43%) and inflammatory disorders (8%). The most common breeds were German Shepherd Dogs, Boxers and Maltese Terriers. Cytopathological and histopathological diagnoses were in complete agreement in 59% of cases, partial agreement in 29% and disagreement in 12%. Conclusion Benign disorders of growth, vascular disturbances and necrosis were the most commonly diagnosed canine splenic disorders, both cytopathologically and histopathologically. Kelpie crosses presented most frequently for cytopathological examination. German Shepherd Dogs were the most common breed diagnosed histopathologically with haemangiosarcoma. Although cytopathological and histopathological splenic examinations are complementary for diagnosis, this study has shown a high correlation for complete and partial agreement between the two.     

Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas

To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6 kDa band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demostration of CD3+ cells in reptiles using specific antibodies.