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1.
自1975年从动物体内提取肿瘤坏死因子(TNF)以来,许多科学家从哺乳动物及禽类体内提取TNF并对其理化性质、生物学活性进行了不同程度的研究和探讨。而用鸡包涵体肝炎病毒诱生TNF与该病之间相关性及产生的消长规律的研究尚属空白。本研究应用鸡包涵体肝炎病毒(FAV-HA毒株)接种SPF雏鸡,在接种后的不同日龄采血,提取TNF,并对其免疫活性(对L929细胞的杀伤率)进行了检测,从而进一步研究其产生TNF的消长规律及与该病毒的相关性。研究结果表明,鸡包涵体肝炎病毒经口腔感染SPF雏鸡可刺激机体产生TNF,在感染的不同日龄鸡的TNF活性出现一定的消长规律。这进一步说明鸡包涵体肝炎(IBH)与TNF的产生及其活性具有密切的相关性。  相似文献   

2.
应用卡介苗(BCG)和大肠杆菌内毒素(脂多糖,Lipopolysuccharide,LPS)鸡体内诱导产生肿瘤坏死因子(Tumor Necrosis Factor,TNF),然后从血清、脾细胞、外周血白细胞提纯TNF,并对其免疫活性(对L929细胞的杀伤作用)进行探讨,将其结果绘出图像,并用方差分析法进行了数据处理。研究结果表明,从鸡体内可以提取TNF。血清TNF活性最强,其次为脾细胞TNF,外周血白细胞TNF活性最弱。血清TNF经用Sephadex G-200分离组分,第一组分活性为最强,但低于全组总活性。  相似文献   

3.
为了制备非洲猪瘟病毒(African swine fever virus,ASFV)p30蛋白卵黄抗体,研究卵黄抗体的生物活性。试验构建了p ET-32a-p30原核表达载体,制定了科学的免疫程序和样本采集方法。提取卵黄液,进行二次盐析,得到对应的卵黄抗体。用酶联免疫吸附试验(enzyme-linked immunosorbentassay,ELISA)测定卵黄抗体效价,用免疫印迹(Westernblotting)技术对其生物活性进行鉴定。ELISA试验表明:首次免疫p30蛋白7d后,能够在蛋鸡的血清中检测出抗体(Ig Y),14d后在卵黄中能够提取到Ig Y,35d后Ig Y效价达到最高值。IgY具备和哺乳动物产生的IgY不同的生物活性,并可具有较高的特异性和稳定性,在免疫学研究中具有重要的价值。利用Ig Y进行实验检测,具有突出优势。通过抗原即ASFV p30蛋白免疫蛋鸡,分离提纯免疫蛋白,检测其生物活性,可用于鉴定非洲猪瘟。  相似文献   

4.
抗菌肽是生物体内诱导产生的具有生物活性的小分子多肽,在哺乳动物体内主要有Defensions和Cathelicidins两大类,抗菌肽PG属于Cathelicidins家族,具有较强抗菌及快速杀菌活性。作者主要综述了抗菌肽PG的结构、基因表达及其机理,以及影响因子和应用前景。  相似文献   

5.
抗菌肽是生物体内天然免疫的重要组成部分,具有广谱抗菌,抗病毒,抗肿瘤等重要生物活性,同时抗菌肽耐酸、耐碱、耐高温、不易产生耐药性等特点使其广泛应用于畜禽生产中,本文从抗菌肽的来源、分类、生物活性、作用机制和在畜禽生产中的应用及展望5个部分进行综述。  相似文献   

6.
抗菌肽是生物体内诱导产生的一类具有抗菌作用的生物活性肽,是哺乳动物防御体系的一个重要组成部分,具有分子量小、热稳定性好、无免疫原性、抗菌谱广等特点,在机体抵抗病原入侵方面起着重要作用。本文就抗菌肽的结构特点、作用机理、应用研究、研究展望及存在问题等方面进行了综述。  相似文献   

7.
目前,绿原酸的诸多功效已被深入研究,其相关产品也相继问世,其在饲料添加剂行业中的应用也逐渐被认可,但其结构类似物——异绿原酸的功能研究与产品开发依旧较少。异绿原酸类化合物亦普遍存在于天然植物体内,其生物活性与绿原酸也大多类似,比如抗氧化、抗菌、抗病毒等。现将近年来异绿原酸的生物活性及提取工艺研究进展进行总结,以期为异绿原酸相关产品的研发提供帮助。  相似文献   

8.
《中国兽医学报》2014,(12):1926-1930
以秀丽隐杆线虫N2野生型为宿主,将鼠伤寒沙门菌ATCC13311定植于线虫体内,然后使用梯度浓度递增法提高培养液中的环丙沙星浓度,诱导沙门菌在线虫体内产生耐药性,同时进行体外诱导耐药试验。对体内、外诱导耐药菌的gyrA、gyrB、parC和parE基因的氟喹诺酮耐药决定区(Quinolone resistance determining regions,QRDR)和外排泵抑制子基因(acrR、marR、ramR和soxR)进行PCR扩增、测序和分析。结果表明,鼠伤寒沙门菌ATCC13311定植于线虫体内,经过诱导后得到环丙沙星MIC为4 mg/L的耐药菌TN4,体外诱导试验获得环丙沙星MIC为4μg/mL的耐药菌TW4。TN4的gyrA基因发生了Asp87Asn突变。体外诱导耐药菌TW4的gyrA基因发生了Ser83Phe和Asp87Val突变,ramR基因出现了20bp的缺失。本研究建立了鼠伤寒沙门菌-秀丽隐杆线虫体内耐药性诱导模型,并比较了体内、外诱导耐药菌的基因突变差异,为进一步研究细菌在动物体内的耐药机理奠定了基础。  相似文献   

9.
应用卡介苗(BCG)和内毒素(LPS)在鸡体内诱导产生肿瘤坏死因子(TNF),检测TNF在pH值5.3、7.0、8.0和在60℃、65℃、70℃各处理30min,胰蛋白酶处理30min,对L929细胞的杀伤活性。结果表明,鸡TNF在pH7.0时活性最强,其次为pH5.3,pH8.0时活性最弱。60℃、65℃各处理30 min活性不受影响,70℃和胰蛋白酶处理30 min活性丧失。  相似文献   

10.
为进一步研究猪IL-10及其生物活性,本研究将猪IL-10基因克隆至原核表达载体p GEX-4T-1,并成功获得其重组蛋白。用IPTG诱导重组蛋白表达,通过GST-亲和柱层析对重组蛋白进行纯化,将纯化的重组蛋白处理猪肺泡巨噬细胞,显著抑制PRRSV感染诱导的IL-12表达,将重组蛋白免疫家兔以制备抗血清,然后利用双向琼脂扩散方法、Western Blot试验对抗血清效价、特异性等生物活性进行了检测。结果表明,该抗血清具有较高的抗体效价和很好的特异性,可以作为IL-10特异性抗体用于今后的试验研究。  相似文献   

11.
Serum tumor necrosis factor (TNF) activity was quantitated in 8 horses given an IV infusion of endotoxin (0.03 micrograms of lipopolysaccharide/kg of body weight, from Escherichia coli 055:B5) in 0.9% NaCl solution over 1 hour. Serum TNF activity was likewise measured in 6 horses given only 0.9% sterile NaCl solution at the same rate. The duration of serum TNF activity was determined, and serum TNF activity was correlated with clinical and laboratory changes during the induced endotoxemia. Horses had no serum TNF activity prior to endotoxin administration, but geometric mean serum TNF activity was significantly higher from 1 to 4 hours after the start of the infusion. In response to endotoxin, horses seemed depressed, had signs of mild to moderate abdominal pain, developed tachycardia and fever, and had leukopenia followed by leukocytosis. Association between serum TNF activity and temperature, heart rate, attitude abnormality score, and WBC count of horses given endotoxin was significant. Serum TNF activity had a significant positive linear correlation with attitude abnormality and heart rate and a negative linear correlation with the WBC count during endotoxemia. Geometric mean serum TNF activity peaked approximately 1.5 hours prior to mean peak fever, and these data were significantly correlated. Results of this study suggest that TNF is an important mediator of endotoxemia in horses.  相似文献   

12.
Tumor necrosis factor (TNF) activity in the circulation of several animal species was determined by a bioassay, using the murine cell line L929. In healthy adult cattle, horses, pigs and dogs, species specific differences of systemic TNF activity were visible. In cattle, TNF activity in the circulation increased during growing up from calf to adult animal. In cattle suffering from various diseases, unchanged, elevated, but also reduced systemic TNF activity have proved to possess clinical relevance. Low systemic TNF activity frequently occurs during lethal inflammatory diseases and may be an indicator of generalized monozyte paralysis.  相似文献   

13.
Local and systemic activation of coagulation is frequently associated with bacterial sepsis. The coagulopathy is due, at least in part, to expression of tissue factor (TF) by monocytes and macrophages. The purpose of this study was to evaluate the expression of procoagulant activity by bovine alveolar macrophages, leukocytes and platelets, and to determine the relative potency of three chemical inhibitors of TF expression (pentoxifylline, retinoic acid, and cyclosporin A). Bovine alveolar macrophages were stimulated with lipopolysaccharide (LPS) derived from Pasteurella haemolytica or recombinant bovine tumour nervous factor (TNF) and dose- and time-dependent effects on TF expression were studied. LPS and TNF induced TF expression in alveolar macrophages and LPS treatment of whole blood induced TF expression in mononuclear cells. Neutrophils and platelets also expressed procoagulant activity, but this activity was not inhibited by anti-bovine TF monoclonal antibody. Pentoxifylline (40 mol/L), retinoic acid (0.01 mmol/L) and cyclosporin A (0.08 mol/L) inhibited TF expression when added concurrently with LPS or TNF, but not when added 4 h after stimulation. TF mRNA was not detected in unstimulated alveolar macrophages by Northern blot analysis. In contrast, exposure to LPS or TNF for 6 h induced marked expression of TF mRNA, which was inhibited by treatment with pentoxifylline, retinoic acid and cyclosporin A. Expression of TNF by alveolar macrophages stimulated with LPS was also inhibited by these compounds. Our results indicate that procoagulant activity expressed by alveolar macrophages and monocytes is associated with expression of TF, whereas procoagulant activity expressed by neutrophils and platelets is not. The concentrations of pentoxifylline and retinoic acid necessary for inhibition of TF expression in vitro may not be achievable in vivo owing to their toxic effects. However, the in vitro concentration of cyclosporin A that inhibited TF expression did not exceed the plasma concentration observed in humans, and therefore may be useful for inhibition of TF expression in vivo.Abbreviations BAL bronchoalveolar lavage - LPS lipopolysaccharide - cDNA cloned deoxyribonucleic acid - cAMP cyclic adenosine monophosphate - GAPDH glyceraldehyde phosphate dehydrogenase - mRNA messenger ribonucleic acid - TF tissue factor - TNF tumour necrosis factor - DPBS Dulbecco's phosphate-buffered saline  相似文献   

14.
Tumor necrosis factor-alpha (TNF) is a pleiotropic cytokine with profound and broad ranging effects on many cell types. There have been few publications investigating the role of TNF in spontaneous disease processes of dogs, particularly the role of this cytokine during endotoxaemia, shock and multiple organ dysfunction syndromes. Plasma samples taken at presentation from 60 dogs with spontaneous acute pancreatitis of varying severity levels (scored 0-4 in ascending severity) were assessed for TNF activity by bioassay and total TNF protein levels through a dot-blot immunoassay. TNF activity by bioassay was detected in 31% (4/13) of dogs presenting with severe disease (>50% expected mortality) as defined using a scoring system for organ compromise, and was not detectable in the remaining animals or healthy controls. TNF activity was detected in 66% (4/6) animals in the highest severity group (Score 4), these animals were showing severe multiple organ dysfunction. Total TNF protein levels, measured by dot-blot immunoassay, exhibited a wide range in all severity groups and healthy dogs. Dogs with detectable TNF activity were not distinguished from the other severity or healthy groups by immunoassay. The absence of detectable differences in total TNF protein levels between the various severity groups suggests that other factors may be crucial in determining the role of TNF in spontaneous canine acute pancreatitis and subsequent endotoxaemia and shock.  相似文献   

15.
本试验旨在建立急性应激致禽类肝损伤的试验动物模型.对麒麟鸡(卷羽鸡)进行持续高温应激后,检测血清中的肝损伤酶谷丙转氨酶(ALT)、天冬氨酸转氨酶(AST)、胆碱脂酶(CHE)和乳酸脱氢酶(LDH)活性的改变,观察肝脏病理组织学变化.结果显示,高温应激前期,麒麟鸡血清肝损伤酶AST和ALT活性极显著升高(P <0.01),高温应激后期,AST活性显著低于正常水平(P <0.05),而ALT活性极显著低于正常水平(P <0.01);持续高温应激期间,CHE的活性一直低于正常水平,高温应激后期,CHE活性极显著低于正常水平(P <0.01);在高温应激下LDH活性呈先下降后上升再下降的变化趋势,高温应激后期,LDH活性极显著高于正常水平(P <0.01).高温应激引起麒麟鸡肝细胞出现水泡变性、肝细胞核浓缩现象.结果表明,采用急性热应激可诱导麒麟鸡肝损伤试验模型,试验模型具备肝脏水泡变性和核浓缩病理特征.  相似文献   

16.
Weaning is characterized by intestinal inflammation, which is a big challenge in pig industry. Control of intestinal inflammation is important for improvement of growth performance and health. Therefore, the study was focused on the anti‐inflammatory activity of low‐molecular‐weight chitosan oligosaccharide (LCOS) in a porcine small intestinal epithelial cell line (IPEC‐J2). The results showed that TNF‐α, as inflammation inducer, significantly upregulated the mRNA expression of IL‐8 and MCP‐1. Afterwards, LCOS significantly attenuated mRNA expression of IL‐8 and MCP‐1 induced by TNF‐α in the cells. Mannose (MAN), as ligand of mannose receptor, had no effect on the anti‐inflammatory activity of LCOS, which suggested that mannose receptor may not involve in the anti‐inflammatory activity of LCOS in IPEC‐J2 cells. Interestingly, N‐[2‐(p‐bromocinnamylamino)ethyl]‐5‐isoquinolinesulfonamide 2HCl hydrate (H89), as PKA (protein kinase A)‐specific inhibitor, reversed the mRNA expression of IL‐8 when co‐cultured with LCOS. Furthermore, LCOS concentration dependent downregulated the mRNA expression of claudin‐1 compared with TNF‐α treatment. However, the trans‐epithelial electric resistance (TEER) was not affected by LCOS when co‐cultured with TNF‐α in 3 hr. In conclusion, LCOS have a potent anti‐inflammatory activity, and as a feed additives, may be useful for the inhibition of inflammatory process in weaning period of pigs with intestinal inflammation occurring.  相似文献   

17.
采取皮下注射硫酸锌,诱导家兔组织产生锌-金属硫蛋白(Zn-MT)。结果:诱导期间肝组织中Zn-MT逐步升高,后期增幅减慢,最后高达5.38mg/g湿组织重;诱导后肾、血、肺和肠中Zn-MT的含量也有较大升高,兔肉中Zn-MT升高幅度最小且含量最低,为0.14mg/g湿组织重。实验表明:除了兔肝以外,肾、血、肺和小肠等组织也可用作Zn-MT提取和纯化的原料。  相似文献   

18.
采取硫酸锌皮下注射家兔,诱导家兔组织产生锌-金属硫蛋白(Zn-MT),诱导期间肝组织中Zn-MT逐步升高,后期增幅减慢,最后高达5.38 mg/g湿组织重;诱导后肾、血、肺和肠中Zn-MT的含量也有较大升高,兔肉中Zn-MT升高幅度最小且含量最低,为0.14 mg/g湿组织重。实践表明:除了兔肝以外,肾、血、肺和小肠等组织也可用于Zn-MT提取和纯化的原料。  相似文献   

19.
A study was performed to determine prevalence of tumor necrosis factor (TNF) activity in serum of equine neonates with presumed sepsis and to determine correlation between serum TNF activity and severity and outcome of disease. Twenty foals less than 21 days old were considered suitable for inclusion in this study by satisfying clinical and laboratory criteria suggestive of septicemia. At admission, blood samples were collected from all foals for determination of serum TNF activity, then clinical course and outcome of disease were recorded. Thirty-one clinically normal foals less than 21 days old served as controls for serum TNF activity. Serum TNF activity was estimated by use of an in vitro cytotoxicity bioassay and WEHI 164 clone-13 murine fibrosarcoma cells. Of the 20 foals with presumed sepsis, 5 had high serum TNF activity. Mean heart rate (P less than 0.005), mucosal petechial hemorrhages (P = 0.06), and death rate (P = 0.06) were greater in the group of foals with high serum TNF activity. These foals also had a lower mean neutrophil count (P less than 0.001), greater band-to-segmented neutrophil ratio (P less than 0.0001), and more prevalent neutrophil toxic changes (P = 0.07) than did foals without serum TNF activity (P = 0.02). Joint swelling was more prevalent in foals without serum TNF activity. Results of the study indicate that serum TNF activity is correlated with clinical criteria of sepsis in equine neonates. An association was apparent between disease severity and serum TNF activity in this group of foals with presumed septicemia.  相似文献   

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