Establishment of proliferative cell nuclear antigen gene as an internal reference gene for polymerase chain reaction of a wide range of archival and fresh mammalian tissues.

Polymerase chain reaction (PCR) from paraffin-embedded tissues provides a powerful tool to amplify DNA from a variety of recent and archival material. Because DNA from paraffin-embedded samples is more degraded than from fresh material, the amplification of reference genes is essential to exclude false-negative results. This study describes the use of the proliferative cell nuclear antigen (PCNA) gene as a reference gene in a range of animal species and in humans. The PCNA-PCR to amplify a fragment extending from exon 5 through exon 6 and including the intervening intron 6 gave a reproducible pattern, with a 280-base pair (bp) band from canine, equine, bovine, ovine, and caprine samples showing high sequence homology. Porcine, guinea pig, tiger, and lion samples, however, gave an additional fragment of approximately 197 bp. The whole intron 6 from these fragments is missing, possibly representing a pseudogene. In feline samples only the 197-bp fragment could be detected. This study shows that the PCNA gene is highly conserved across a broad range of animal species and is well suited as an internal control for PCR analysis in veterinary medicine.     

遮阴对10种野生观赏植物生长及生理特性的影响

为了探讨野生观赏植物在遮阴条件下的生态适应性及寻求适度的遮阴比例,选取兰州地区10种野生观赏植物为试验材料,比较测定不同光强生境(全光照、25%、50%、70%遮光率)对10种野生观赏植物生长和生理生化的影响。结果表明,角茴香(Hypecoum erectum)和驴蹄草(Caltha palustris)含水率随着遮阴度的增加而增加,小花草玉梅(Anemone rivularis)在75%遮阴度时干重最大,显著高于其余遮阴处理及全光照(CK)(P0.05);藤本铁线莲(Clematis florida)可溶性糖含量随着遮阴度的增加而减少,鹅绒委陵菜(Potentilla anserina)、角茴香、水杨梅(Geum aleppicum)和驴蹄草的叶绿素含量随着遮阴度的增加而增加;东方草莓(Fragaria orientalis)、角茴香和驴蹄草在全光照下丙二醛的含量显著高于其它处理(P0.05),香青(Anaphalis sinica)和水杨梅在75%遮阴度下丙二醛含量较高。综上所述,香青、莓叶委陵菜(Potentilla chinensis)、藤本铁线莲和野棉花(A.vitifolia)比较适合全光照生长;鹅绒委陵菜、小花草玉梅、东方草莓、角茴香和水杨梅可以忍耐适当的遮阴环境;驴蹄草适合在遮阴条件下生长。     

鸭疫里氏杆菌免疫原性研究Ⅰ.荚膜的形态学观察及荚膜物质的提取

本研究应用电镜对Ⅰ型鸭疫里氏杆菌的荚膜进行了初步观察,在电镜下,负染的菌体周围呈一电子透明层;未经抗体处理的细菌超薄切片,荚膜在菌体周围呈松散的絮状结构,而应用同源抗体处理后,荚膜则呈一均匀层。同时在超薄切片中还看到细菌的空泡和外膜结构。采用十六烷基三甲基溴化铵（ＣＴＡＢ）成功地提取了培养物上清液中的荚膜成分,经过测定比较,加入５ｍＭＣＴＡＢ时效果最佳。     

内蒙古草原N2O和CH4排放通量及其季节变化特征研究

１９９８年５－１２月,作者在内蒙古半干旱原用静态箱法进行原位Ｎ２Ｏ和ＣＨ４排放通量观测,分析了Ｎ２Ｏ、ＣＨ４排放量及其季节变化和相关过程,定时估计草原Ｎ２Ｏ和ＣＨ４收支在人类活动下的变化。重点讨论了内蒙古古草原型群落羊草,大针茅和退化草原Ｎ２Ｏ和ＣＨ４的排放通量及其季节变化特征。     

Pharmacokinetics of sanguinarine,chelerythrine, and their metabolites in broiler chickens following oral and intravenous administration

Sanguinarine (SA) and chelerythrine (CHE) are the main active components of the phytogenic livestock feed additive, Sangrovit®. However, little information is available on the pharmacokinetics of Sangrovit® in poultry. The goal of this work was to study the pharmacokinetics of SA, CHE, and their metabolites, dihydrosanguinarine (DHSA) and dihydrochelerythrine (DHCHE), in 10 healthy female broiler chickens following oral (p.o.) administration of Sangrovit® and intravenous (i.v.) administration of a mixture of SA and CHE. The plasma samples were processed using two different simple protein precipitation methods because the parent drugs and metabolites are stable under different pH conditions. The absorption and metabolism of SA following p.o. administration were fast, with half‐life (t_1/2) values of 1.05 ± 0.18 hr and 0.83 ± 0.10 hr for SA and DHSA, respectively. The maximum concentration (C_max) of DHSA (2.49 ± 1.4 μg/L) was higher that of SA (1.89 ± 0.8 μg/L). The area under the concentration vs. time curve (AUC) values for SA and DHSA were 9.92 ± 5.4 and 6.08 ± 3.49 ng/ml hr, respectively. Following i.v. administration, the clearance (CL) of SA was 6.79 ± 0.63 (L·h^?1·kg^?1) with a t_1/2 of 0.34 ± 0.13 hr. The AUC values for DHSA and DHCHE were 7.48 ± 1.05 and 0.52 ± 0.09 (ng/ml hr), respectively. These data suggested that Sangrovit® had low absorption and bioavailability in broiler chickens. The work reported here provides useful information on the pharmacokinetic behavior of Sangrovit® after p.o. and i.v. administration in broiler chickens, which is important for the evaluation of its use in poultry.     

利用耳组织、血液、牛奶样品检测牛病毒性腹泻病毒的比较

牛病毒性腹泻病以发热、口腔及消化道粘膜糜烂、腹泻为主要症状,可引起怀孕母牛流产或产畸型胎儿,给养牛业造成巨大的经济损失,该病已成为当前严重危害我国养牛业的传染病之一。由于大部分感染BVDV的牛不表现特征性的临床症状和病理变化,而且引起腹泻和消化道黏膜糜烂或溃疡的疾病很多,所以确诊需进行实验室检查。目前实验室检测BVDV的主要方法是利用美国IDEXX公司ELISA诊断试剂盒,通过BVDV抗原和抗体的检测区别牛群持续感染牛。用于检测BVDV的样品种类有许多种,绝大多数是采集血液(血清)或牛奶样品进行检测。本试验对奶牛进行耳组织采样,同时采集血液和牛奶,用三种样品在相同条件下同时检测BVDV抗原,用血清和牛奶样品检测抗体,比较样品的检测结果。结果表明,这三种样品检测结果具有一致性,且耳组织采样更宜于大面积筛查。     

传染性喉气管炎病毒WG株糖蛋白gJ基因的克隆与表达

根据GenBank中已发表的传染性喉气管炎病毒全基因组序列（NC006233）设计引物,以传染性喉气管炎病毒WG株基因组为模板,PCR扩增糖蛋白gJ基因,并对其进行序列分析。用DNAStar软件分析酣蛋白的抗原性,选择抗原性强的140aa-483aa片段,克隆并连接到原核表达载体pET-32a（＋）中,转化BL21（DE3）菌株。经IPTG诱导后,获得大小为65．2ku的重组蛋白,命名为卜出,表达量占菌体蛋白总量的60．8％;Westernblot分析表明r-gJ具有较强的抗原性。     

不同生境下陌上菜的形态解剖学比较

本研究首次采用GMA切片等方法对不同生境下陌上菜的形态结构进行了深入的研究。结果显示,陌上菜种苗发育具有水生植物的特征:种子及种苗需浸水萌发及发育,具下胚轴毛,胚根发育迟缓。成株的营养器官兼具水生及陆生植物的结构特征:完全陆生的植株和湿生植株的气生部位具有较厚角质层,发达的机械组织及较发达的通气组织。完全水生的植株和湿生植株浸水部位的角质层和机械组织不发达,但通气组织非常发达。生殖器官:完全陆生和湿生的植株具有闭锁和开放2种类型的花,而完全水生的植株仅具闭锁花,从而保证了这一物种在不同生境下均能正常结实。此外,陌上菜具有特立中央胎座。本研究首次揭示了陌上菜在不同生境下种苗发育、营养器官及生殖器官结构的变化,完善了陌上菜的形态学,为进一步研究母草科的系统学提供了形态学依据。     

规模猪场母猪繁殖障碍综合征的病因调查

作者采用9套试剂盒，检测了7种能引起猪繁殖障碍综合征的传染病，对来自有繁殖障碍症状的305场次进行猪瘟(HC)抗原检测，并对包括这305场次在内的978个猪场(次)病例的6346份血清样品及52698份田间血清样品进行HC、猪繁殖与呼吸综合征(PRRS)、伪狂犬病(PR)、日本乙型脑炎(JE)、细小病毒感染(DPV)、猪衣原体病(Chla)、布鲁氏菌病(Bruc)等7种传染病的抗体检测。结果表明，在有繁殖障碍症状的猪场，HC抗原检出率高达61．97％，并且在不使用疫苗的情况下，PR3RS、PR、JE、Chla和PPV的抗体阳性场分别为49．42％、34．29％、12．72％、31．71％、和48．08％。HC和：PR交叉感染率达23．81％；HC和：PRRS交叉感染率为9．52％，PR和PRRS交叉感染率高达59．65％；另外JE、PPV也同HC、PR、PRRS存在部分交叉感染。不使用疫苗的田间血清样品的PRRS、PR的抗体阳性率也高。HC与PRRS．PR、Chla、JE和PPV中的一种或几种混合感染可能是引起繁殖障碍造成严重损失的主要原因。加强综合防制，优化免疫程序、把握引种关、加强生物安全措施是防制猪繁殖障碍综合征的关键。