成年小鼠雄性生殖细胞的冷冻保存

在含10％小牛血清（NBS）的DMEM培养基中，分别各添加5％，10％，15％，20％和25％的二甲基亚砜（DMSO）,丙二醇（PG），乙二醇（EG）和甘油（G），对成年小鼠睾丸生殖细胞冷冻保存；复苏后台盼蓝染色测定细胞复苏率。结果显示，5％－25％DMSO冻存液组的细胞复苏率分别为88.5%,88.0%,65.6%及51.3%;5%-25%PG冻存液组的细胞复苏率分别为87.2%,86.4%,79.0%,73.4%及40.1%.;5%-25%EG冻存液组的细胞复苏率分别为6.6%,80.9%,60.8%,51.3%及30.0%;5%-25%G冻存液组的细胞复苏率分别为86．5％，86．3％，65．3％，36．0％及31．4％。其中各抗冻剂5％和10％组的细胞复苏率最高，与15％组相比均存在显著或极显著差异。4种抗冻剂的最高细胞复苏率之间无显著差异。DMSO，PG，EG和G分别冷冻保存成年小鼠睾丸生殖细胞对的最小损失率分别为4．8％，6．1％，6．7％，6．8％。结果表明，采用慢速冷冻时，DMSO，PG，EG及G均适宜用作成年小鼠睾丸生殖细胞的抗冷冻剂，最佳使用含量均为5％－10％。成年小鼠睾丸生殖细胞分别在含5％－10％的DMSO，PG，EG和G的DMEM（含10％NBS）冻存液中，2步慢速降温，液氮储存，37℃水浴复苏，是一种具有较高复苏率的冷冻保存方法。     

家蚕血液、消化液中酸性核糖核酸酶和天冬酰胺酶活性的初步研究

测定了家蚕夏芳、长灰A和大造各品种血液、消化液中酸性核糖核酸酶及家蚕秋白、夏芳、长灰A和大造各品种天冬酰胺酶性变化。结果表明：各品种血液、消化液中酸性核糖核酸酶、天冬酰胺酶活性变化随发育呈现较强的规律性，血液中，4龄第3d、5龄第3d这两种酶均出现峰值，4龄眠前活性较低，消化液中，5龄第3d这两种酶活性均较高，不同品种这两种酶活性峰值出现时间有差异；各品种血液中酸性核糖核酸酶、天冬酰胺酶性均明显高于相应品种消化液中这两种酶活性；各品种之间血液、消化液中这两种酶活性有明显差异。     

羔羊轮状病毒的诊断与防治研究

用分离培养的羔羊轮状病毒进行了高免血清、荧光抗体、酶标记抗体、RNA核酸电泳等的制造和研究,对我省羔羊轮状病毒鉴定为亚群Ⅰ,长型.用高免血清治疗羔羊、犊牛轮状病毒性腹泻疗效显著.初步认定羔羊轮状病毒与犊牛轮状病毒之间有交互抗原.对分离到的病毒进行了长期的细胞培养继代,并经过较长时间的低温处理,经用初生羔羊口服免疫,试验结果表明,弱毒疫苗对初生羔羊具有较好的免疫力.     

梅花鹿铜缺乏综合征的复制

为了证明梅花鹿铜缺乏综合征的发生与饲料铜含量的关系，以含铜３．３３×１０－６的日粮饲喂１３１只幼龄圈养梅花鹿，于２８个月后开始发生铜缺乏综合征，继续饲喂到３９个月时，其显症率达９．１６％。人工复制的梅花鹿铜缺乏综合征的临床症状与自然发生者相一致，主要为患鹿后躯运动失调，渐进性瘫痪；意识清楚，无癫痫、抽搐症状发生；被毛颜色变淡，眼睛周围出现白眼圈。ＲＢＣ为（７．５６±０．８３）×１０１２／Ｌ，Ｈｂ（１６３．３０±１２．９０）ｇ／Ｌ，血清铜蓝蛋白氧化酶（２５．７３±６．４５）ＩＵ／Ｌ，均显著低于对照组。血清铜（５．６２±０．８０）μｍｏｌ／Ｌ，毛铜（４．５５±１．２８）ｍｇ／ｋｇ，均显著低于对照组；血清铁（２８．４８±２．４７）μｍｏｌ／Ｌ，显著高于对照组。主要病理变化为脑、脊髓神经脱髓鞘；部分神经元变性坏死；血管壁肥厚，内膜粗糙，管腔变窄，脆性增加；肝、脾、淋巴结含铁血黄素沉着及骨质疏松。日粮中铜含量为７．３３×１０－６时，梅花鹿发育良好，体质健壮，生产性能优越，无铜缺乏综合征发生。表明这一铜含量可作为梅花鹿铜元素饲养标准的参考值。     

复合酶对肉仔鸡生产性能及代谢的影响

选取１日龄ＡＡ肉仔鸡８０只，随机分成二组，研究日粮添加和不添加复合酶对肉仔鸡生产性能、代谢及免疫的影响。结果表明：复合酶使肉仔鸡日增重提高７％～７．７％，料肉比降低了４．５％～４．９％，但采食量增加不显著；血液中尿酸和血糖浓度显著下降，血浆胰岛素与血清Ｔ３浓度显著提高，提示复合酶改善了肉仔鸡营养物质的吸收与代谢；此外，血清蛋白和血浆ＩｇＧ水平高于对照组，表明复合酶促进了免疫功能     

A dominant antigenic epitope on SARS-CoV spike protein identified by an avian single-chain variable fragment (scFv)-expressing phage

Severe acute respiratory syndrome (SARS) is a newly emergent human disease, which requires rapid diagnosis and effective therapy. Among antibody sources, immunoglobulin Y (IgY) is the major antibody found in chicken eggs and can be used as an alternative to mammalian antibodies normally used in research and immunotherapy. In this study, phage-expressing chicken monoclonal scFv antibody was chosen and characterized with phage display antibody technology. Truncated fragments of SARS-CoV spike protein were cloned in pET-21 vector and expressed in BL-21 Escherichia coli (E. coli) cells. After purification, the purity of these recombinant spike proteins was examined on SDS-PAGE and their identity verified with Western blot analysis using anti-his antibodies and sera from convalescent stage SARS-CoV-infected patients. Using these bacteria-derived proteins to immunize chickens, it was found that polyclonal IgY antibodies in the egg yolk and sera were highly reactive to the immunogens, as shown by Western blot and immunocytochemical staining analysis. A phage displaying scFv library was also established from spleen B cells of immunized chicken with 5 x 10(7) clones. After four panning cycles, the eluted phage titer showed a 10-fold increase. In sequence analysis with chicken germline gene, five phage clones reacted, with large dissimilarities of between 31 and 62%, in the complementarity-determining regions, one dominant phage 4S1 had strong binding to fragment Se-e, located between amino acid residues 456-650 of the spike protein and this particular phage had significantly strong binding to SARS-CoV-infected Vero E6 cells. Based on the results, we conclude that generating specific scFv-expressing phage binders with the phage display system can be successfully achieved and that this knowledge can be applied in clinical or academic research.     

黑茶镳子扦插育苗技术研究

对3个品种黑茶镳子‘坠玉’‘寒丰’‘绥研’的扦插技术研究表明,以当年生枝条为穗材,‘坠玉’用ABT 500 mg/L速蘸,生根率达43.3%,生根数达8~10条;‘寒丰’用ABT 200 mg/L或IBA 200 mg/L浸泡3 h,扦插生根率达100%,生根数大于30条;‘绥研’用ABT 500 mg/L速蘸3~5 s,扦插生根率达83.3%,生根数达8~10条。     

油菜素内酯提高蔬菜作物抗逆性的研究进展

油菜素内酯化合物(Brassinosteroids,BRs)是国际上公认的一类高效、广谱、无毒的新型植物甾醇类生长激素。近年来研究发现,BRs的使用可显著提高植物对生物和非生物逆境的抗性,从而促进植物生长发育,大幅度提高作物产量。本文综述了BRs的类型、应用方式以及在提高蔬菜抵抗干旱、低氧、温度、盐离子、病害、重金属等环境胁迫方面的研究成果,并对未来的研究方向进行了展望。     

Expression of Jagged2/Notch3 signaling molecules in pulmonary hypertensive rats induced by monocrotaline

AIM: To study the expression of Jagged2/Notch3 signaling molecules in pulmonary vascular wall of pulmonary hypertensive rats induced by monocrotaline. METHODS: SD rats were randomly divided into normal control group (C group,n=15), solvent control group (S group,n=15) and monocrotaline model groups (M group,n=15). The model of pulmonary hypertension was established by a single subcutaneous injection of monocrotaline (50 mg/kg). The rats in S group were given a single subcutaneous injection of the same dose of solvent. After 4 weeks, the pulmonary vascular remodeling was assessed by HE staining, and the mean pulmonary artery pressure (mPAP) and right ventricular systolic pressure (RVSP) were determined by right heart catheterization. The expression of Jagged2/Notch3/Hes5 molecules in the pulmonary vascular wall was detected by immunohistochemical method and real-time PCR. RESULTS: Compared with S group and C group, the percentage of medial wall thickness of smaller arteries in model group increased significantly (P<0.01). The levels of mPAP and RVSP in M group were significantly higher than those in S group and C groups (P<0.01). The results of real-time PCR showed that the expression of Jagged2, Notch3 and Hes5 was significantly increased in M group compared with S group and C group. The data from immunohistochemical detection indicated that Jagged2 mainly expressed in the intima of small lung artery, Notch3 and Hes5 mainly expressed in the medial smooth muscle cells. Compared with S group and C group, the expression of Jagged2 and Notch3 was significantly increased in the lung small arteries of M group. CONCLUSION: The activation of Jagged2/Notch3 signaling pathway might play an important role in the formation of pulmonary hypertension.