Immunohistochemical analysis of pseudorabies virus spread through neurons innervating the eyeball

Pseudorabies virus (PRV) was inoculated intraocularly into BALB/c mice, and the distribution pattern of cells positive for several neurotransmitters and viral antigens in the eyeball, trigeminal nerve ganglia, and superior cervical ganglia was examined immunohistochemically to clarify the neural route of the virus spread. In the eyeball, substance P (SP)- and calcitonin gene-related peptide (CGRP)-positive cells were detected in the ipsilateral iris and ciliary body, neuropeptide tyrosine (NPY)-positive cells were detected in the choloid membrane, and tyrosine hydroxylase (TH)-positive cells were detected in the ipsilateral inner nuclear layer of the retina; all these cells contained viral antigens. In the superior cervical ganglia, viral antigen-positive cells containing TH or NPY were found at bilateral sites. In the trigeminal nerve ganglia, viral antigen-positive cells containing SP or CGRP were found at bilateral sites. These findings indicated that the SP- and CGRP-positive ganglion cells of the trigeminal nerve ganglia innervating the iris or ciliary body, and the NPY-positive ganglion cells of the superior cervical ganglia innervating the iris, ciliary body, and choroid membrane served as the route for the virus spread. These findings also suggested that dopaminergic neurons, such as the TH-positive retinal cells and TH-positive ganglion cells of the superior cervical ganglia, served as the route for virus spread.     

用麦蛾卵大量繁殖甘蓝夜蛾赤眼蜂(Trichogramma brassicae)

在2000年建成的麦蛾生产线基础上,经多次试验后完成了具有日生产赤眼蜂200万头生产能力的繁蜂设备,2001年最高日繁蜂量120万头。通过改进生产工艺和设备,扩大规模,提高繁蜂效率,日繁甘蓝夜蛾赤眼蜂1100万头,生产能力提高了5.5倍。生产成本比原来降低了47.3％。     

Concentrations and specific antibodies to staphylococcal enterotoxin-C and toxic shock syndrome toxin-1 in bovine mammary gland secretions,and inflammatory response to the intramammary inoculation of these toxins

To investigate the pathological role of staphylococcal enterotoxins (SEs) and toxic shock syndrome toxin-1 (TSST-1) in bovine mastitis, the production of SEs and TSST-1 was investigated in staphylococci isolated from 120 mammary gland secretions (MGS, 51 from no clinical sign-mammary glands and 69 from staphylococcal mastitic-mammary glands) collected from dairy farms where staphylococcal mastitis frequently occurred in Miyagi and Yamagata prefectures from 1997 to 1998. Concentrations of these toxins and specific antibody titers in each MGS were also measured. Furthermore, SEC and TSST-1 were inoculated into lactating mammary glands and inflammatory responses were analyzed. A high percentage of staphylococci including Staphylococcus aureus and coagulase-negative staphylococci isolated from both no clinical sign- and mastitic-MGS produced both SEC and/or TSST-1. The concentration of SEC increased with the severity of the mastitis, and was significantly higher (P<0.05) in acute mastitic-than in no clinical signs-MGS. Titers of specific antibodies to TSST-1 in MGS were significantly higher (P<0.05) than those to SEC, regardless of whether or not the cows were lactating or mastitic. Specific antibodies purified from MGS neutralized each toxin in vitro. A significant increase (P < 0.05) in somatic cell counts was induced by the intramammary inoculation of SEC but not TSST-1. These findings indicated that SEC rather than TSST-1 plays an important role in the pathology of staphylococcal bovine mastitis. The inflammatory activity of TSST-1 was probably neutralized by specific antibodies in MGS.     

Experimental infection of recent field isolates of feline herpesvirus type 1

Two field isolates of feline herpesvirus type 1 (FHV-1) designated as 00-015 and 00-035, were obtained from cats diagnosed as feline viral rhinotracheitis (FVR) in Japan. To analyze the character of recent FHV-1, these two isolates and our laboratory strain C7301 were inoculated experimentally to specific-pathogen-free cats. Although all cats showed typical FVR symptoms, more severe clinical symptoms were observed on cats infected with the isolates 00-015 and 00-035 compared with those of C7301-infected cats. Severe ocular lesions including conjunctivitis were found in the cats infected with the isolates, indicating that the recent FHV-1 has a potential to induce severe FVR symptoms including ocular lesions.     

Sequence analysis of the genes encoding the phosphoprotein of recent isolates of canine distemper virus in Japan

The nucleotide sequences of the phosphoprotein (P) of canine distemper virus (CDV) strains isolated between 1992 and 1996 in Japan were determined. This is the first report of the complete sequences of the P genes of recently prevalent CDV strains. The deduced amino acid sequences of the P, C and V proteins showed that in the new Japanese isolates, these proteins have approximately 93%, 90-91% and 92% identities with those of the Onderstepoort vaccine strain, respectively. The predicted functional regions were conserved. RNA editing resulting in a shift to the open reading frame (ORF) of the V protein was shown to occur with the same efficiency in both the field isolates and vaccine strain.     

日粮色氨酸水平对生长猪的氮利用效率、血浆类胰岛素生长因子-I、生长激素及胰岛素的影响

本试验旨在研究日粮色氨酸水平对生长猪的氮利用率以及对血浆类胰岛素生长因子-I(IGF-I)、生长激素(GH)、胰岛素浓度的影响。试验以4头大白去势公猪作为试验动物,以色氨酸含量分别为1.2、1.7、2.2和2.7g/kg的4种日粮为试验处理,按照4×4拉丁方进行试验设计。结果表明,色氨酸水平对干物质消化率、有机物消化率和氮消化率没有显著性影响(P>0.05)。氮沉积和氮的生物学利用率随着日粮色氨酸水平由1.2提高到1.7、2.2和2.7g/kg而显著提高(P<0.05),但是试验处理1.7、2.2和2.7g/kg之间的氮沉积和氮的生物学价值没有显著差异(P>0.05)。不同处理之间的血浆IGF-I、GH和胰岛素浓度没有显著差异(P>0.05)。日粮色氨酸水平和血浆IGF-I、GH或胰岛素浓度之间以及氮沉积和血浆IGF-I、GH或胰岛素浓度之间均无显著的相关关系(P>0.05)。综合以上结果,日粮色氨酸水平对生长猪的氮沉积具有显著影响,而对血浆IGF-I、GH和胰岛素没有显著影响。     

鱼精蛋白的研究现状及应用

鱼精蛋白(protam ine)又称精蛋白,是一种主要存在于各类雄性动物成熟精巢组织中的多聚阳离子肽,它与DNA紧密结合在一起,以核精蛋白的形式存在。鱼精蛋白是一种小而简单的球形碱性蛋白,一般由30~50个氨基酸组成,其中以精氨酸为主。目前,已从多种鱼类(如鲑鱼、红鳟鱼、鲱鱼等)和哺乳动物(如老鼠、人、野猪等)的成熟精巢组织中提取到了鱼精蛋白。根据氨基酸的种类和数量,鱼精蛋白可分为3种:①单鱼精蛋白,它的碱性氨基酸仅为精氨酸;②双鱼精蛋白,碱性氨基酸除含有精氨酸外,还含有组氨酸或赖氨酸;③三精蛋白,碱性氨基酸有精氨酸、组氨酸和赖氨…     

高盐对肉鸡肺小动脉中膜平滑肌细胞表型转化的影响

观察了高盐诱发肉鸡腹水综合征（AS）发生发展过程中肺小动脉中膜平滑肌细胞（SMC）表型转化的规律，旨在探讨肺小动脉中膜SMC表型转化在AS发生过程中的可能作用。100只1日龄商品代AA雄性肉仔鸡常规育雏，8日龄起随机分为对照组和高盐组（饮水中添加0．30％氯化钠）。分别在15、22、29、36、43、50日龄时每组随机取5只鸡的肺组织并制备石蜡切片，以免疫组织化学方法结合图像分析法测定SMC表型标志蛋白α-SM-actin的表达。结果表明：（1）对照组肉鸡的生长发育过程中存在肺小动脉中膜SMC表型的动态转化，22日龄时发生了由收缩表型向增殖表型的转化，随后逐步向收缩表型回转，且各级肺小动脉SMC转化时间不一致。（2）高盐组SMC表型转化时间（15日龄）较对照组提前了1周，高盐促进了SMC由收缩表型向增殖表型转化。且直径20～50μm的肺小动脉SMC表型转化受高盐影响最大，推测其在高盐诱发肉鸡AS的过程中可能起更重要的作用。     

造血和神经相关标志物在人胎盘的表达

为探讨人胎盘组织在造血中的作用以及研究造血和神经标志物在胎盘中的表达和在胚胎发育中的相关性，用免疫组织化学法对人胎盘组织和体外培养的胎盘组织中的贴壁细胞进行染色，观察其造血因子和神经标志物的表达。结果发现人7月龄胎盘组织血管内血细胞表达多种造血细胞因子，如SCF、VEGF、BMP-4和FGF-1和KDR，血管内皮细胞上也有表达；胎盘组织同时表达造血干细胞标记CD34、CD133和神经细胞标记物Nestin和MAP2等。而足月龄胎盘CD34、CD133、KDR和造血相关因子SCF、VEGF、BMP-4、FGF-1等弱表达，不表达Nestin和MAP2。两组胎盘切片都不表达GFAP和MBP。胎盘贴壁细胞（hPDACs）碱性磷酸酶、波形蛋白、CD133、Nestin和MAP2染色呈阳性，CD34、GFAP和MBP阴性表达。人胎盘组织和胎盘贴壁细胞表达多种造血相关因子和神经细胞标志物，提示胎盘具有造血功能，造血形成和神经发生之间可能存在基因表达叠加现象。     

缺氧与肉鸡肺动脉平滑肌细胞c-fos和 c-myc基因表达的关系

利用细胞培养、原位杂交、免疫组化、图像分析等技术研究缺氧与肉鸡肺动脉平滑肌细胞癌基因c-fos和c-myc表达的关系,进一步阐明缺氧与以肺动脉增殖重塑为特征的肉鸡腹水综合征的关系。结果显示,缺氧显著引起肺动脉平滑肌细胞内癌基因c-fos和c-mycmRNA的表达(c-fosmRNA:常氧组为144.6±20.2,缺氧组为198.1±32.8,P<0.01;c-mycmRNA:常氧组为125.4±18.8,缺氧组为167.1±22.4,P<0.01)。缺氧显著引起肺动脉平滑肌细胞内癌基因c-fos和c-myc蛋白的表达(c-fos蛋白:常氧组为150.9±33.2,缺氧组为225.9±37.0,P<0.01;c-myc蛋白:常氧组为162.1±28.5,缺氧组为228.8±33.4,P<0.01)。结果表明缺氧能够明显诱发肉鸡肺动脉平滑肌细胞内癌基因c-fos、c-myc的转录和表达,是启动肺动脉平滑肌细胞增殖的重要原因。本研究阐明缺氧是以肺动脉重塑为特征的肉鸡腹水综合征的主要诱因。