中国禽(番鸭)呼肠孤病毒YB株S4基因序列分析

应用非免疫番鸭胚增殖中国禽(番鸭)呼肠孤病毒YB分离株,用LSTRIAZOL提取病毒RNA,反转录-聚合酶链反应(RT-PCR)扩增中国禽(番鸭)呼肠孤病毒YB分离株S4基因节段cDNA.将S4cDNA克隆到PMDl8-T载体上,并进行了鉴定和核苷酸序列测定.序列分析结果,克隆的S4cDNA共1 124bp,包括非编码区和完整的阅读框架.分子进化系统分析表明该毒株与禽呼肠孤病毒(鸡呼肠孤病毒)的亲缘关系较远,DRV-YB与DRV-89330同缘率为93.3%.     

Metabolism of [ring-2,6-14]parathion in plant cell suspension cultures of carrot (Daucus carota), purple foxglove (Digitalis purpurea), soybean,thorn apple (Datura stramonium) and wheat (Triticum aestivum)

By means of standardized procedures, the metabolism of [ring-2,6-¹⁴C]-parathion was investigated in carrot (Daucus carota L.), purple foxglove (Digitalis purpurea L.), soybean (Glycine max Merrill cv. ?Mandarin’?, and Glycine max Merrill cv. ?Harosoy 63’? cultivated on B5 and Miller media, respectively), thorn apple (Datura stramonium L.), and wheat (Triticum aestivum L.) cell suspension cultures. In the wheat and soybean (Mandarin) cells only 2.9 and 8.9%, respectively, of the applied parthion remained unmetabolized after 48 h of incubation, while 51.2, 57.9, 60.3, and 62.4% of the unchanged parent were detected in the D. purpurea, D. Stramonium, carrot and soybean (Harosoy) cultures, respectively. In all suspensions, paraoxon and 4-nitrophenol were found as phase I metabolites, thus demonstrating that plant tissues can catalyse oxidative desulfuration and dearylation of parathion. 4-Nitrophenol was also glycosylated with glucose and possibly galactose. Further, as yet unidentified, metabolites indicated that bio-transformations had also occurred at the aromatic moiety. Large amounts of non-extractable residues were detected in the wheat suspension (38.3%), while the other cultures showed a lower incorporation of ¹⁴C into insoluble cell material (0.9-9.4%). For a prospective ecotoxicological evaluation of the metabolic fate of pesticides and xenobiotics in plants in general, the differential metabolic capacity of plant cell cultures and plants should be taken into account.     

Development of novel pesticides based on phytoalexins: Part 2. Quantitative structure-activity relationships of 2-heteroaryl-4-chromanone derivatives

Phytoalexins are low-molecular-weight chemicals that immune systems of plants produce and accumulate in response to infections, especially those of fungal origin. Although their content is not high in plants, yet they have shown unique fungicidal activity and played an important role in the defence system of plants. In searching for novel environmentally benign fungicides with high activity, the structures of flavanone derivatives, one of the most important phytoalexins groups, have been modified via bioisosteric substitution and a series of 2-heteroaryl-4-chromanones were designed and synthesized. They showed good fungicidal activities against rice blast disease, Pyricularia grisea (Sacc). Their IC50 values were tested in vitro and the relationship between structure and fungicidal activity was analyzed quantitatively using a Hansch-Fujita approach. The results showed that hydrophobicity was very important for fungicidal activity and there is apparently an optimum hydrophobic property for the molecules at a log Pow value of about 2.7. In addition, the results indicated that electronic effects played an important role in binding with the receptor and that the C=O group was probably a electron-accepting site. The quantitative structure-retention correlative equation of the title compounds was also established.     

三江源区高寒草甸草场放牧牦牛采食量的测定

试验选用约2．5岁公牦牛8头，在三江源地区玉树县的高寒草甸草场，应用4N盐酸不溶灰分法（4N—AIA）和木质素标记物法测定了不同物候期（青草期和枯黄期）放牧牦牛日采食量及牧草营养成分的消化率。结果表明：2．5岁牦牛在青草期、枯黄期的100kg体重日采食量分别为3．69kg和2．96kg，差异极显著（P〈0.01）；其不同物候期的千物质消化率分别为65．70％、61．72％（P〉0．05），粗蛋白的消化率为62．25％、39．00％（P〈0．01），粗脂肪为49．36％、58．18％（P〈0．05），ADF的消化率为57．88％、62．67％（P〉0．05），NDF的消化率为69．09％、79．38％（P＜0．01）。     

Characterization of polymorphisms of transferrin receptor and their association with susceptibility to ETEC F4ab/ac in pigs

Diarrhoea caused by enterotoxigenic Escherichia coli (ETEC) expressing F4 (F4ab, F4ac and F4ad) fimbriae is a significant cause of mortality and morbidity in newborn and weaned pigs. The locus controlling susceptibility towards ETEC F4ab/ac has been mapped to SSC13q41, in which TFRC (transferrin receptor) was localized and considered as a positional candidate gene for ETEC F4ab/ac receptor. In this study, we determined susceptibility/resistance to ETEC F4ab/ac in a total of 755 F2 animals from a White Duroc x Erhualian intercross using a microscopic enterocyte adhesion assay. We identified two TFRC polymorphisms (SNPs 591 A>G and 632 A>G) in a single exon after comparative sequencing analysis of 2371-bp amplicons containing the complete coding region of TFRC using RNA of eight full-sib F2 animals with susceptible and resistant phenotypes. The intron sequences flanking the two exon polymorphisms were obtained, revealing an intron polymorphism (SNP 291 C>T). We genotyped the 19 founder animals of the White Duroc x Erhualian intercross for the identified polymorphisms, showing that only the 291 C>T polymorphism is a highly informative marker. We further genotyped all 59 F1 and 755 F2 animals for the 291 C>T polymorphism, and the association of this polymorphism with susceptibility/resistance to ETEC F4ab/ac in these F2 animals was evaluated by the transmission disequilibrium test. The result showed that the 291 C>T polymorphism is not a causal mutation, however, has a significant linkage disequilibrium with the ETEC F4ab/ac, especially F4ac receptor locus.     

Characterization of a line of pigs previously selected for increased litter size for RBP4 and follistatin

The objective of this study was to determine if selection response for increased litter size in pigs could be partially attributed to three type 1 marker loci coding for genes known to affect litter size: oestrogen receptor (ESR), retinol‐binding protein 4 (RBP4) and follistatin (FS). In the high litter size line (LS), pigs from the largest litters, based on number of pigs born alive (NBA), were retained to parent the next generation. A randomly selected control line (LC) was maintained. Gilts were reared in litters of 10 pigs or less to minimize maternal effects. Pigs were measured at generations 10–12. Additional traits scored were number of fully formed pigs (NFF) and number of mummified fetuses (MUM). Breeding values for NFF and NBA were greater (p < 0.05) in LS than LC in generations 11 and 12, but no significant line differences were found for MUM. The A allele of the ESR locus was fixed in both lines. After adjustment for effects of genetic drift, frequency of the two alleles segregating for the FS and RBP4 loci did not differ significantly between lines. No significant additive or dominance effects of the FS markers were detected for NFF, NBA and MUM in either LS or LC. Response to selection for increased litter size could not be attributed to effects at the ESR, RBP4 or FS loci.     

新城疫La Sota、V4疫苗免疫鸡和免疫攻毒鸡的特异性IgA抗体动态变化规律比较

应用间接ELISA对新城疫LaSota、V4疫苗免疫鸡及免疫攻毒鸡的IgA抗体动态变化进行了测定和比较。试验表明，LaSota和V4免疫鸡泪液中特异性IgA均在免疫后第5天出现，8天开始明显升高，与血清中IgG出现时间相似，免疫后21天达到高峰。LaSota免疫鸡的泪液IgA抗体水平略高于V4免疫鸡；而两免疫组哈德氏腺(HG)中的特异性IgA高峰出现较迟。免疫攻毒鸡泪液中的特异性IgA抗体水平均先呈短暂的升高，之后下降的趋势，而HG中的IgA则首先表现降低，之后很快升高，5天后趋于下降，两攻毒组差异不显著，对IgA回忆应答均不明显。     

中国大耳白肉兔IFN-γ、IL-2和IL-4基因的克隆测序与进化分析

实验应用反转录-聚合酶链式反应(RT-PCR)技术,在国内首次从ConA诱导培养的中国大耳白肉兔外周血淋巴细胞总RNA中扩增得到IFN-γ、IL-2和IL-4基因,并将其克隆到PGEM-T载体中,经菌落PCR鉴定、序列测定及序列分析,结果表明:①经克隆得到的IFN-γ基因(序列号:DQ852341),与Genbank中已登录的欧洲兔IFN-γ基因(序列号:AB010386)的核苷酸序列和推知氨基酸序列的同源性均为100%;与其他哺乳动物如犬、猫、人、猪、牛、羊、马、大熊猫和鼠等的核苷酸同源性在63.0%(鼠)～77.6%(人)之间;编码氨基酸同源性在41.7%(鼠)～65.7%(人)之间;②扩增得到的IL-2基因(序列号:DQ852342),与Genbank中已登录的欧洲兔IL-2基因(序列号:AF068057)的核苷酸序列和推知氨基酸序列的同源性分别为99.6%和100%;与其他哺乳动物如犬、猫、人、猪、牛、羊、马、大熊猫和鼠等的核苷酸同源性在64.8%(鼠)～86.0%(人)之间;编码氨基酸同源性在55.7%(鼠)～80.1%(人)之间;③扩增得到的IL-4基因(序列号:DQ852343),与Genbank中已登录的欧洲兔IL-4基因(序列号:AF169169)的核苷酸序列和推知氨基酸序列同源性均为100%;与其他哺乳动物如犬、猫、人、猪、牛、羊、马、大熊猫和鼠等的核苷酸同源性在57.0%(大熊猫)～69.8%(人)之间;编码氨基酸同源性在43.0%(鼠)～53.6%(人)之间。用这3个基因分别构建的进化树结果都表明,兔与人的亲缘关系相对较近,与鼠的亲缘关系最远,这与传统的分类地位基本吻合,即兔与鼠应分别归为兔形目和啮齿目动物。     

Oct4在山羊睾丸组织中的表达与定位

探讨不同时期山羊睾丸组织中干细胞的分布和特点,以45 d和75 d山羊胎儿及出生后4月龄山羊睾丸为试验材料,进行了HE染色和Oct4抗体的免疫组化染色。结果显示,在45 d山羊胎儿睾丸组织中睾丸实质细胞聚集呈团状,形成岛屿状组织结构;发育到75 d山羊胎儿睾丸组织中睾丸的间质与实质组织结构差异明显,睾丸生精小管已经形成,生精小管管腔闭锁;出生后4月龄山羊睾丸组织结构明显,生精小管形态特征清晰,但管腔仍处于闭锁状态;Oct4免疫组织化学显示在45 d山羊胎儿睾丸组织中Oct4阳性细胞呈岛屿状分布,多见于生精小管前期的组成细胞;对于75 d山羊胎儿睾丸组织Oct4阳性细胞呈环状分布于生精小管中,睾丸间质组成细胞中则较少;在出生后4月龄山羊睾丸中阳性细胞主要分布于睾丸组织生精小管基底膜,沿基底膜环状分布。试验证实了在早期山羊睾丸组织中Oct4主要表达在生精前体细胞。