大鼠急性冷应激模型的建立

通过分析冷刺激对大鼠血清中细胞因子、皮质酮(CORT)、促肾上腺皮质激素(ACTH)的含量以及外周血淋巴细胞中热休克蛋白70(Hsp70)表达量的影响,构建大鼠急性冷应激模型,旨在为其相关研究提供研究基础。本试验急性冷刺激时间为12h。利用ELISA方法检测各组大鼠血清中IL-2、IL-4、IL-6、IL-10、TNF-α、CORT、ACTH的含量;采用Western blot和qRT-PCR方法检测各组大鼠外周血淋巴细胞中HSP70及mRNA表达量。结果表明,与对照组相比,急性冷刺激组大鼠血清中IL-2和ACTH的含量显著升高(P0.05),IL-4、IL-6、TNF-α和CORT的含量极显著升高(P0.01),IL-10无显著变化(P0.05);外周血淋巴细胞内HSP70及其mRNA表达量呈显著升高水平(P0.05)。成功构建大鼠急性冷应激模型。     

在免疫反应中LI-10相关生物学活性研究进展

<正>细胞因子是指由免疫细胞和某些非免疫细胞经刺激而合成分泌的一类生物活性物质,多属多肽或糖蛋白。细胞因子与内分泌激素不同,多数细胞因子不出现在血清中,而是以自分泌和(或)旁分泌形式发挥作用。1细胞因子产生的两个重要途径(1)Th1分泌的细胞因子有IL-1、IL-2、IL-6、IL-8、IFN-γ、CM-CSF、TNF-α等,参与细胞介导的免疫过程及组织炎症反应,即巨噬细胞活化,淋巴毒素作用,     

皂树皂苷QS21增强猪支原体肺炎活疫苗滴鼻免疫小鼠效果评价

为提高猪支原体肺炎活疫苗的滴鼻免疫效果,以猪支原体肺炎(168株)活疫苗为抗原,以QS21为免疫佐剂混合后滴鼻免疫小鼠,ELISA 法检测免疫效果,包括血清和支气管肺泡灌洗液中抗原特异性抗体水平,并测定了Th1型细胞因子IFN-γ、Th2型细胞因子IL-4和Th17型细胞因子IL-17的分泌情况。结果表明,QS21能激活黏膜和全身性的体液免疫,显著升高血清和支气管肺泡灌洗液中IgG,IgG1、IgG2a抗体含量,并能提高支气管肺泡灌洗液中黏膜免疫抗体sIgA的含量。QS21还能显著增加支气管肺泡灌洗液中IL-4、IFN-γ和IL-17的分泌,激活Th1、Th2和Th17型细胞免疫应答。以上结果表明QS21能全面激活免疫系统,具有开发为猪支原体肺炎活疫苗黏膜免疫佐剂的应用前景。     

3类CpG ODNs正调节猪IP-10基因的表达研究

CpG ODN诱导不同物种先天性免疫反应的分析结果表明,诱导细胞因子(CK)分布的种类和水平存在明显的异同。试验对3类CpG ODNs(A、B、C类)如何激活猪先天性免疫生物标记及猪免疫细胞在体内外培养(含CpGODNs)中的先天性免疫反应动力学进行了研究。利用实时定量PCR(SYBR Green)分析了细胞因子和趋化因子基因的mRNA表达水平。结果发现,A-类CpG ODN可显著诱发IFN-γ、IL-12(P40)、IL-6、IL-4、TNF-αmRNA的瞬间表达;C-类CpG ODN可显著诱发IFN-γ、IFN-α、IL-12 mRNA的表达,并使IL-4(Th-2型)mRNA降至最低表达水平,GCODNs可刺激使一些细胞因子的表达水平降至最低;B-classGpC ODN7909能显著影响IL-12(P35)mRNA表达水平。值得注意的是,12 h时3类CpG ODNs刺激均可显著诱导IP-10的mRNA表达。体内外试验结果表明,IP-10可作为CpG ODNs诱发猪免疫活性的生物标记。     

IP-10作为牛结核病诊断标志物的初步探究

为评价细胞因子IL-12 p40、IP-10和TNF-α转录水平与牛分枝杆菌感染之间的关系,及其在牛结核病诊断中的应用潜力。采集田间筛选的结核病阳性牛、结核病阴性牛以及牛分枝杆菌68002人工感染牛的外周血淋巴细胞,经牛结核菌素(PPDB)、重组蛋白CFP-10-ESAT-6(CE)、MPT63、PET和PBS分别刺激6 h,提取细胞总RNA,用荧光定量PCR检测IL-12 p40、IP-10、IFN-γ和TNF-α的转录水平。结果显示结核病阳性牛的外周血淋巴细胞经PPDB和CE刺激后,其IP-10的m RNA转录水平显著高于结核病阴性牛,且与IFN-γ的m RNA转录水平具有良好的相关性;初步建立牛结核病IFN-γ和IP-10的Real-time PCR检测方法,其对临床阳性样本的检出率分别为71.45%和78.57%。因此,IP-10的m RNA转录水平与牛分枝杆菌的感染相关,有作为牛结核病诊断标志物的潜力。     

紫锥菊提取物对小鼠血清细胞因子的影响

探索紫锥菊提取物对小鼠血清细胞因子的影响。试验选取健康成年小鼠80只,随机分为8组(每组10只),分别为空白对照组(给予生理盐水灌胃)、免疫抑制模型组(腹腔注射环磷酰胺溶液100 mg/kg)、免疫抑制试验组和正常试验组。免疫抑制试验组和正常试验组分别以低、中、高不同剂量紫锥菊提取物(25 mg/kg·d、50 mg/kg·d和100 mg/kg·d)进行灌胃,连续灌胃4周后,测定其血清细胞免疫因子含量。试验结果表明,与空白组相比,免疫抑制模型组的IL-2、IL-6和TNF-α含量均明显降低(P0.01),而不同剂量正常试验组的IL-2、IL-6和TNF-α含量也存在显著差异(P0.05);与免疫抑制模型组相比,紫锥菊低、中、高剂量免疫抑制试验组IL-2、IL-6和TNF-α水平显著升高(P0.05),高剂量免疫抑制试验组小鼠的IL-2、IL-6、TNF-α水平分别升高25.21%、29.24%、67.38%。     

复方中药“猪康散”对断奶仔猪血清中细胞因子水平的影响

选取20日龄健康杜长大三元杂交仔猪40头,随机分为4组,每组10头。27³1日龄时分别在基础日粮中添加0.0%,0.5%,1.0%,2.0%"猪康散"。于35,45,55,65日龄每组随机选取4头仔猪,前腔静脉采血测定IL-2、IL-6、IFN-γ、TNF-α等细胞因子的水平。结果表明,1.0%添加组在各个时间点均可明显提高血清中IL-2、IL-6、IFN-γ和TNF-α的水平,且显著优于其他添加组;这表明在断奶仔猪日粮中添加1.0%"猪康散"能通过促进血清中IL-2、IL-6、IFN-γ、TNF-α的表达量而增强仔猪细胞免疫功能。     

热应激后猪血清中IL-2、IFN-γ及TNF-α水平的动态变化

检测热应激后猪血清中的IL-2、IFN-γ及TNF-α水平,观察其变化规律.将12头体重基本一致的3月龄中国试验用小型猪随机分成高温试验组和常温对照组两组.高温试验组在人工气候仓内饲养,模拟夏季炎热气候,气温从26～39℃24 h循环变温,39℃时维持4 h;常温对照组在22℃下饲养.试验持续10 d,在热应激的第1、3、5、7、9天,39℃高温持续期结束的时刻采血,分离血清.用ELISA法检测各血清样品中IL-2、IFN-γ及TNF-α水平.结果显示：IL-2水平有所上升,且在热应激的第1天和第3天与常温对照组差异显著（P＜0.05）;IFN-γ水平有所下降,且在热应激的第3天和第5天与常温对照组差异显著（P＜0.05）;TNF-α水平有所上升,但与常温组比较差异不显著（P＞0.05）;热应激试验进行的10 d中,IL-2、IFN-γ及TNF-α水平变化有其共同特点,即随着试验的进行各细胞因子浓度都向常温对照组的正常值发展.本研究从细胞因子的角度验证了前人热应激抑制机体免疫机能的结论.     

抗应激复合添加剂对模拟运输应激大鼠血液相关生化指标的影响

将30只清洁级Wistar大鼠随机分为试验组、普通对照组及应激对照组。试验组用自拟抗应激复合添加剂(维生素C+维生素E+碳酸氢钾+γ-氨基丁酸,羧甲基纤维素钠助悬)进行灌胃,2对照组灌胃0.5%羧甲基纤维素钠(CMC)溶液。1周后利用摇床模拟运输应激,检测大鼠血清肌酸激酶(CK)、乳酸脱氢酶(LDH)、血糖(GLU)、促肾上腺皮质激素(ACTH)、糖皮质激素(GC)以及部分细胞因子(IL-2、IL-4、IL-6和TNF-α)等应激血清标志物水平。结果显示,大鼠经2 h模拟运输应激后,各项指标均有不同程度升高,而自拟抗应激添加剂对运输应激引起的CK、ACTH、GC、IL-2、IL-4和IL-6水平升高有明显的抑制作用;表明自拟的抗应激复合添加剂对抵抗大鼠运输应激具有较好的效果。     

Th1/Th2型细胞因子的动态变化对山羊妊娠的影响

为了研究Th1/Th2型细胞因子的动态变化对山羊妊娠的影响,探讨其生理学意义,试验通过EUSA方法对山羊妊娠早期及流产前后外周血液中Th1(TNF-α、IL-2)与Th2(IL-10)型细胞因子的含量进行了检测.结果表明:妊娠早期外周血液中TNF-α和IL-2含量先升高后降低,IL-10含量持续上升;流产后TNF-α和IL-2含量高于流产前含量(P＜0.01或P＜0.05),IL-10则低于流产前含量(P＜0.05或P＜0.01).说明山羊正常妊娠时Th1/Th2型细胞因子动态平衡应当是以Th2型免疫及其相关因子占优势,平衡失调则导致妊娠失败.     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.     

Mechanism of Action of Probiotic Function of Lactobacilli

乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.     

Evaluation of serum concentrations of biomarkers of skeletal metabolism and results of radiography as indicators of severity of osteochondrosis in foals

OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.