野马追克喘散治疗猪呼吸道疾病效果试验

目的：应用野马追克喘散新兽药对呼吸道综合征进行临床疗效试验,为推广野马追克喘散提供科学依据。方法：将自然感染、症状典型、猪呼吸道综合征的140头病猪（杜长大三元杂交）,平均体重20～25kg,随机分为7个组,每组20头,其中第Ⅰ、Ⅱ、Ⅲ组用野马追克喘散治疗,分别以高、中、低剂量给药;第Ⅳ、Ⅴ组分别用麻杏石甘散、金花平喘散以常规剂量给药;Ⅵ、Ⅶ组以硫酸卡那霉素注射液（0.2mL/kg体重）和泰乐菌素（0.4mL/kg体重）以常规剂量给药。结果：野马追克喘散的治疗效果较麻杏石甘散和金花平喘散治疗效果好,治愈率也达93.3%。结论：野马追克喘散治疗猪气喘咳嗽疗效较高,收效较快,维持时间较长,是较理想的药物。     

用RAPD标记研究7个猪种间的亲缘关系

试验利用随机扩增多态ＤＮＡ技术对互助猪、荣昌猪、甘肃白猪、长白猪、约克夏猪、杜洛克猪、汉普猪共７个品种的ＤＮＡ池进行了多态型研究,分析了各品种间的亲缘关系。结果显示：甘肃白猪虽然是由国外品种和地方品种培育而成,但其亲缘关系更接近于中国猪品种;约克夏猪与其它欧洲猪种亲缘关系较远,这可能与约克夏猪在培养过程中亲本遗传背景较为复杂有关。     

卵母细胞去核及卵丘细胞移核方法对猪体细胞核移植的影响

以猪体外成熟卵母细胞周围分离得到的卵丘细胞作为猪体细胞核移植的供核细胞,研究了卵母细胞不同去核方法(盲吸法、活性荧光染色去核法、盲吸法和活性荧光染色法联合去核法)的去核率差异,并比较了卵丘细胞透明带下注核和胞质内注核两种重建胚构建方法的胚胎裂解率、4 8细胞发育率和桑椹胚率的差异。结果表明:(1)以荧光染料染色法的去核率(87.18%)最高,与其他试验组相比差异显著(P<0.05);盲吸法去核所用时间最少(1.2 min/个),与其他试验组相比差异显著(P<0.05)。(2)透明带下注核比卵母细胞质内直接注核的卵裂率(40.11%/29.75%)和4 8细胞率(11.86%/6.81%)高,差异显著(P<0.05);桑椹胚率(1.75%/1.79%)无明显差异(P>0.05)。     

猪圆环病毒2型PCR检测方法的建立及应用

根据Genbank中PCV2全基因组序列设计了扩增片段大小为262bp的引物P1/P2,建立了检测猪圆环病毒2型PCR方法。引物对P1/P2从PCV2阳性病毒感染基因组中扩增出特异性条带的最低DNA含量为10-9ng/mL(约100个PCV2病毒)。用该方法对陕西和甘肃的103份临床发病猪的肺脏和淋巴结样品进行检测,结果有68份样品检测出阳性,阳性率为66%。     

重组GP5AB蛋白间接ELISA检测PRRSV抗体方法的研究

利用GST-GP5AB重组蛋白作为包被抗原,通过反应条件优化,建立了用于检测猪繁殖与呼吸综合征病毒(PRRSV)抗体的间接ELISA方法。抗原最适包被浓度为2μg/mL,最佳封闭液为0.15%BSA,37℃封闭2 h后,再4℃封闭24 h,血清最适稀释度为1∶200,其作用时间为60m in,酶标抗体最适稀释度为1∶20 000,最适作用时间为90 m in,37℃显色10 m in,S/P≥0.284为阳性,S/P≤0.26为阴性,介于二者之间为可疑的判定标准。该抗原与猪其他4种临床症状类似的疾病的阳性血清反应呈阴性。批内和批间重复性试验结果,变异系数均小于7%,表明本方法具有较好的特异性和重复性。应用本方法初步检测了一些疫苗免疫仔猪血清样品,并与重组N蛋白和PRRSV抗原同时进行比较,结果显示3种抗原检测的结果基本一致。     

猪繁殖与呼吸障碍综合征病毒基因芯片检测技术的建立与优化

根据猪繁殖与呼吸综合征病毒（PRRSV）的核衣壳蛋白编码基因序列设计了一对特异性引物P1／P2。扩增出大小为294bp的目的片段；再针对这个基因片段。设计合成4条寡核苷酸探针。其中反向引物的5’端用荧光素Cy3标记。以荧光标记不对称PCR技术为基础。通过将单链PCR产物与芯片杂交实现对PRRSV的检测。建立PRRSV的基因芯片检测方法。利用该方法对39份猪组织样品进行检测。与RT—PCR检测方法相比。本方法具有良好的特异性和敏感性。试验结果表明用该方法快速检测病料组织中PRRSV是可行的，对该病的进行快速诊断和分子流行病学调查具有重要意义。     

猪圆环病毒复制与转录模式研究进展

猪圆环病毒(PCV)是一种单股、环状DNA病毒,有两种血清型,其中PCV-2是引起断奶仔猪多系统衰竭综合征的重要病原,给世界养猪业造成了巨大的经济损失.文章主要从PCV-1和PCV-2的基因组结构、复制起始区的结构与功能、复制相关蛋白的结构与功能、转录图谱、各转录物的剪接方式及主要转录物的转录效率等方面进行了综述,同时也对二者的复制与转录模式的异同点进行了比较.这些结果对PCV-1和PCV-2毒力差异的分子机制研究具有一定的借鉴意义,同时可为抗PCV药物的研发提供一定的理论依据.     

Porcine surfactant in treatment of LPS-induced early-stage acute lung injury in rats

AIM: To evaluate the therapeutic efficacy of intratracheal instillation of porcine pulmonary surfactant (PPS) in rats with lipopolysaccharides (LPS)-induced early-stage ALI in this study.METHODS: SD rats weighing 200 g-300 g were randomly divided into 4 groups: LPS (1.5 mg·kg-1)+saline,LPS+PPS 100 mg·kg^-1,LPS+PPS 150 mg·kg^-1,LPS+PPS 200 mg·kg^-1.The PaO₂ and PaCO₂,as well as survival rate of rats were examined for 6 h after the start of PPS-instillation.Then,rats were killed and lungs were immediately removed for lung index (LI) and histological analysis.The bronchoalveolar lavage fluid (BALF) was collected for measurement of total protein (TP) contents,TNF-α level and white blood cell(WBC) numbers.RESULTS: Significantly increased PaO₂,reduced mortality rate,decreased total protein and TNF-α contents in BAL,as well as lung index and meliorated histological appearance were observed in three PPS-treated groups compared with group given saline after LPS (P<0.05).The therapeutic effect in PPS150 and PPS200 groups was better than that in PPS100 group.CONCLUSION: Intratracheal PPS instillation provides protective effect on acute lung injury in rats induced by LPS.     

Report on the analyses of mAb reactive with porcine CD8 for the second international swine CD workshop

Based on an analysis of their reactivity with porcine peripheral blood lymphocytes (PBL), only three of the 57 mAbs assigned to the T cell/activation marker group were grouped into cluster T9 along with the two wCD8 workshop standard mAbs 76-2-11 (CD8a) and 11/295/33 (CD8b). Their placement was verified through the use of two-color cytofluorometry which established that all three mAbs (STH101, #090; UCP1H12-2, #139; and PG164A, #051) bind exclusively to CD8⁺ cells. Moreover, like the CD8 standard mAbs, these three mAbs reacted with two proteins with a MW of 33 and 35 kDa from lymphocyte lysates and were, thus, given the wCD8 designation. Because the mAb STH101 inhibited the binding of mAb 76-2-11 but not of 11/295/33, it was given the wCD8a designation. The reactivity of the other two new mAbs in the T9 cluster with the various subsets of CD8⁺ lymphocytes were distinct from that of the other members in this cluster including the standards. Although the characteristic porcine CD8 staining pattern consisting of CD8^low and CD8^high cells was obtained with the mAb UCP1H12-2, a wider gap between the fluorescence intensity of the CD8^low and CD8^high lymphocytes was observed. In contrast, the mAb PG164A, not only exclusively reacted with CD4⁻/CD8^high lymphocytes, but it also failed to recognize CD4/CD8 double positive lymphocytes. It was concluded that this mAb is specific for a previously unrecognized CD8 epitope, and was, thus, given the wCD8c designation. A very similar reactivity pattern to that of PG164A was observed for two other mAbs (STH106, #094; and SwNL554.1, #009). Although these two mAbs were not originally positioned in the T cell subgroup because of their reactivity and their ability to inhibit the binding of PG164A, they were given the wCD8c designation. Overall, five new wCD8 mAbs were identified. Although the molecular basis for the differences in PBL recognition by these mAbs is not yet understood, they will be important in defining the role of CD8⁺ lymphocyte subsets in health and disease.     

Analyses of monoclonal antibodies reacting with porcine CD3: results from the Second International Swine CD Workshop

Among the 57 monoclonal antibodies (mAb) analyzed within the T-cell group from the Second Swine CD Workshop, six mAb fell within clusters T10 and T11 (No. 088, STH164; No. 148, FY1A3; No. 149, FY2C1; No. 150, FY1H2; No. 151, FY2A11; No. 169, BB23-8E6). The mAb within these two groups gave a similar appearance on flow cytometry and stained all peripheral blood T-cells as defined by CD4 and wCD8 staining. All six mAb precipitated a 24 kDa protein. On the basis of inhibition analyses performed as part of the workshop and from published data, the mAb define at least three epitopes. There is only minimal stimulation of resting peripheral lymphocytes, but four of the mAb produce strong stimulation in the presence of PMA. With the exception of STH164, all have been shown to react with CD3-transfected COS cells. The new mAb, therefore, react with three epitopes on porcine CD3 designated CD3a (BB23-8E6, FY2A11), CD3b (FY1A3, FY2C1), and CD3c (FY1H2). mAb STH164 appears to be reactive with another epitope, however, since its reactivity with CD3 has not been confirmed it is designated as wCD3.