High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

五种鬼伞过氧化物酶和酯酶的同工酶研究

应用垂直板聚丙烯酰胺凝胶电泳对五种野生鬼伞 (Coprinus)真菌进行了过氧化物酶 (POD)和酯酶(EST)的同工酶分析 ,结果表明 :五种鬼伞的POD和EST同工酶酶谱比较稳定清晰 ,且分别有一条共同的酶带 ,可能是鬼伞属的POD和EST同工酶特征酶带 ;POD和EST同工酶酶谱均表明 ,家园鬼伞 (C .domesticus)和瓦鳞鬼伞 (C .clavatus)间有较近的亲缘关系 ;不同种鬼伞的POD和EST同工酶之间既有共同的特征 ,又各自有本物种的特有特征 ,POD和EST同工酶酶谱可以作为鬼伞属种类鉴定、亲缘关系比较的重要依据。     

Roles of the Kv1.2, Kv1.5, Kv2.1 potassium channels in hypoxia pulmonary vasoconstriction

AIM: To determine the role of Kv1.2, Kv1.5, Kv2.1 in the hypoxia pulmonary vasoconstriction (HPV). METHODS: Male Wistar rats were divided into two groups: normoxic group and hypoxic group. The single smooth muscle cell was obtained from pulmonary artery of Wistar rats with acute enzymatic digestion method. The conventional whole-cell patch clamp technique was used to record the resting membrane potential (Em) and the potassium currents of voltage-gated potassium channel (IKv) in rat pulmonary arterial smooth muscle cells (PASMC). Intracellular application of Kv1.2/Kv1.5/Kv2.1 antibodies (1∶125) was conducted through the whole-cell patch clamp system. RESULTS: ① Em of PASMC was depolarized after 24 h hypoxia compared with that of control cells . IKv of PASMC was decreased after 24 h hypoxia, . ② The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies depolarized Em and inhibited IKv in PASMC from normoxic rat, whereas the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on them. ③ The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies and the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on IKv and Em from rats hypoxic for 24 h. CONCLUSION: Kv1.2, Kv1.5, Kv2.1 might be oxygen sensitive potassium channels which mediated HPV.     

In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network

Development of new semen cryopreservation techniques improving sperm survival and ensuring availability of viable spermatozoa for a prolonged time‐period after AI is promising tools to reduce sensitivity of timing of AI and enhance overall fertility. The SpermVital^® technology utilizes immobilization of bull spermatozoa in a solid network of alginate gel prior to freezing, which will provide a gradual release of spermatozoa after AI. The objective of this study was to compare post‐thaw sperm quality and in vitro sperm survival over time of Norwegian Red bull semen processed by the SpermVital^® (SV) technology, the first commercialized production line of SpermVital^® (C) and by conventional procedure applying Biladyl^® extender (B). Post‐thaw sperm motility was not significantly different between SV, C and B semen (p > .05). However, sperm viability and acrosome intactness were higher for SV than C and B semen (p < .05). Small differences in DNA quality were observed (p < .05). Sperm viability after storage in uterus ex vivo was higher for SV than for C semen (p < .05). Furthermore, sperm survival in vitro over time at physiological temperature was significantly higher for SV semen than C semen as well as B semen during the incubation period of 48 hr (p < .05). In conclusion, the SpermVital^® technology is improved and is more efficient in conserving post‐thaw sperm quality and results in higher sperm viability over time in vitro for SV than for C and B semen.     

池塘和稻田养殖模式下克氏原螯虾肌肉和肝脏营养成分比较

通过对池塘和稻田2种养殖模式下克氏原螯虾(Procambarus clarkii)的肌肉常规营养成分进行测定,同时,对肌肉和肝胰腺的氨基酸和脂肪酸组成进行测定,从而对其营养品质进行分析评价.结果显示,2种养殖模式下克氏原螯虾粗蛋白和粗脂肪无显著性差异(P＞0.05),池塘养殖模式下水分显著高于稻田模式(P＜0.05),...     

大黄鱼消化道菌群结构、消化酶和非特异性免疫酶活力分析

为研究大黄鱼(Larimichthys crocea)消化道的菌群结构、消化酶和非特异性免疫酶活力特征,本研究采用高通量测序技术系统分析大黄鱼胃、幽门盲囊和肠道中菌群组成及分布,并对比研究工厂化养殖和网箱养殖模式下的消化道菌群;同时,结合生化分析方法解析2种模式下消化道消化酶和非特异性免疫酶活力特征。结果显示,2种养殖模式下,菌群多样性随消化道延伸呈下降趋势;乳杆菌科(Lactobacillaceae(f))、Fructobacillus、黄杆菌属(Flavobacterium)等代表的菌属为共有优势菌群。其中,拟杆菌属(Bacteroides)和Anaerostipes等的丰度随消化道延伸呈下降趋势,而乳杆菌科、E01_9C_26_marine_group(o)所代表的菌属及黄杆菌属等则相反;普氏菌属(Prevotella_9)、乳杆菌科代表的菌属为2种模式养殖大黄鱼的主要差异菌属。工厂化养殖条件下,幽门盲囊和肠道中的菌群组成及其参与营养和免疫相关代谢通路的基因数目差异不显著(P>0.05),但与胃部的菌群组成和相关代谢通路基因数目存在明显差异;而网箱养殖大黄鱼胃部与幽门盲囊和肠道的菌群结构及相关代谢通路基因数目差异相对较小。2种养殖模式下的大黄鱼消化道菌群与饲料菌群相近。另外,胃和幽门盲囊也具有非特异性免疫酶活性,说明,整个消化道还具有一定的化学免疫屏障作用。本研究结果将为大黄鱼健康养殖提供基础参考,并为消化道菌群生理功能探讨提供理论依据。     

次生幼龄林封山育林后乔木层物种结构变化研究

1999年在浙江省仙居县俞坑村的封山育林区设置20 m×20 m的固定样地2块,对连续5年的调查数据进行统计分析,以其对次生幼林乔木物种结构变化进行研究.分析结果,乔木层物种丰富度指数增加了7～11,乔木物种增加了2～5科,3～8属.自然恢复为以壳斗科、杉科、山茶科、樟科、胡桃科、安息香科等物种为建群种或共建种的次生常绿阔叶林、针阔混交林.     

干草防腐剂在不同规格苜蓿干草捆中的应用效果研究

为了研究干草防腐剂的防霉效果,本文在调制苜蓿小方捆时添加巴斯夫防腐剂BLP,并在苜蓿大方捆中比较使用了自行研发的防腐剂LZP和防腐剂BLP。结果表明：在苜蓿小方捆中,25%水分打捆时,添加防腐剂BLP显著抑制了苜蓿草捆中霉菌的生长,但对营养品质没有显著影响;30%水分打捆时,防腐剂BLP显著降低了苜蓿干草中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)的含量,当添加量为6 L·t^-1时,防腐剂BLP的防霉效果较为显著且增加了粗蛋白(CP)的含量。在苜蓿大方捆中,16%～19%水分打捆添加防腐剂BLP和LZP均能很好抑制霉菌的生长;而22%～25%水分打捆时时,两种防腐剂的对霉菌的抑制效果较差。上述结果表明调制苜蓿干草小方捆时添加6 L·t^-1防腐剂BLP能有效抑制霉菌的生长;自行研发的防腐剂LZP与BLP的防霉效果类似,具有一定的应用前景,但仍需进一步证明。     

绿鳍马面鲀雌雄性腺转录组比较分析

为探明绿鳍马面鲀(Thamnaconus septentrionalis)性腺发育相关基因表达特征,优化亲本生殖调控技术,本研究对绿鳍马面鲀雌雄亲体的精巢和卵巢进行了转录组测序分析,经de novo拼装,最终获得119 219个单基因(unigene),N50长度为1 255 bp。在NR、NT、KO、SwissProt、PFAM、GO及KOG数据库分别注释到24 009、35 057、18 453、26 971、30 294、11 420和21 613个unigene。绿鳍马面鲀精巢和卵巢转录组中存在18 954个差异表达基因(DEG),相对于精巢,在卵巢中上调表达的基因有11 265个,下调表达的有7 689个。选取bmp2、sox3、figla、hsd17b1、cyp19a、cyp17、foxl2、star和amh 9个DEGs进行实时荧光定量PCR (qRT-PCR)验证,结果显示,qRT-PCR结果与RNA-Seq分析相一致。GO和KEGG富集结果分析发现,amh、cyp17和star可能在绿鳍马面鲀雄性精子发生过程中起关键作用;bmp2、foxl2、cyp19a、figla和hsd17b1在雌性卵子发生和卵巢类固醇生成过程中发挥重要作用。本研究通过比较绿鳍马面鲀精巢和卵巢的转录组表达差异,初步阐明了精巢和卵巢的基因表达特征,为进一步研究绿鳍马面鲀的性腺发育机制奠定了基础。     

农业科研机构专业技术人员绩效考核体系的研究

针对绩效考核的理论研究和发展趋势,借鉴同行经验,分析了上海市农业科学院实施绩效考核实践的经验和存在问题。对绩效考核体系提出了改进意见,便于更准确的甄别优劣,激励先进、鞭策落后,帮助科技人员确切了解工作的不足,及时调整方向、改进工作。