荷斯坦牛红细胞Na~+-K~+-ATP酶的研究进展

作者对荷斯坦牛红细胞Na+-K+-ATP酶的分子结构和基本特征、分布、作用机制、调控机制、生理功能、影响因素及其与热应激的关系进行了综述.     

氮磷钾配施对甘草育苗质量的影响

利用“3414”方案进行N、P、K营养液配方设计,在配方营养液浇灌条件下进行甘草育苗质量比较,旨在探寻适宜甘草育苗的施肥配方,为其测土配方施肥和无土栽培育苗提供依据。结果表明,以纯沙石作为育苗基质,在添加Hoagland营养液条件下,不同N、P、K配比营养液对经98%硫酸处理的甘草种子的出苗质量和幼苗生长发育均具有极显著影响。N₂K₂配比下出苗质量均优于空白对照,且随着施磷水平的提高出苗质量呈下降趋势,出苗质量依次为N₂P₀K₂＞N₂P₁K₂＞N₂P₂K₂＞N₂P₃K₂。而出苗活力依次为N₂P₁K₂＞N₂P₀K₂＞N₂P₂K₂＞N₂P₃K₂。N₂P₂K₂和N₂P₁K₂处理均可极显著提高甘草根系活力,为培育壮苗奠定良好基础。单纯缺N、缺K和过量N、P和K肥均不利于甘草出苗和幼苗的生长发育。以上说明在甘草出苗阶段需P量小,但出苗后开始显示其营养效应。从育苗效益和环境保护双重考虑,配方施肥宜采用N₂P₁K₂,即添加80 mg/L NH₄NO₃、89 mg/L KH₂PO₄和298 mg/L KNO₃作为N、P和K营养源,每隔20 d浇1次。需要注意的是浇灌后应及时喷灌适量水,使氨态氮下渗到底部基质,以避免氨的挥发而灼伤幼苗。     

MAP3K5基因在畜禽剩余采食量表型调控中的研究进展

随着分子遗传学的发展,已经鉴定出了影响剩余采食量(RFI)的大量数量性状位点和候选基因。有丝分裂原活化蛋白3激酶5(MAP3K5),也称凋亡信号调节激酶1(ASK1),属于MAPK超家族基因之一。目前,已有细胞外信号调节蛋白激酶(ERK)、c-Jun N-末端激酶(JNK)和p38丝裂原激活蛋白激酶(p38-MAPK)这3个MAPK家族成员在哺乳动物细胞中被克隆和鉴定,其主要作用机制是介导3条MAPKs信号通路,从而影响家畜的生长、体型及产奶性状等。课题组在前期对RFI的研究中筛选出与牛剩余采食量相关的MAP3K5基因,但其功能作用尚不明确,笔者在此基础上回顾了该基因的结构、生物学功能,概述了该基因在主要畜禽采食量变异及人类肥胖表型中的功能及作用,并从遗传学的角度重点分析了MAP3K5基因在畜禽RFI表型调控中的可能机制。通过对MAP3K5基因在畜禽RFI表型调控中的研究进展进行综述,期望为后期深入开展MAP3K5基因在畜禽采食量性状调控中的分子机制研究提供思路;对于其他可能通过MAP3K5基因影响畜禽表型的因素(如肠道菌群)有待进一步探讨。     

Cell-free extract from porcine induced pluripotent stem cells can affect porcine somatic cell nuclear reprogramming

Pretreatment of somatic cells with undifferentiated cell extracts, such as embryonic stem cells and mammalian oocytes, is an attractive alternative method for reprogramming control. The properties of induced pluripotent stem cells (iPSCs) are similar to those of embryonic stem cells; however, no studies have reported somatic cell nuclear reprogramming using iPSC extracts. Therefore, this study aimed to evaluate the effects of porcine iPSC extracts treatment on porcine ear fibroblasts and early development of porcine cloned embryos produced from porcine ear skin fibroblasts pretreated with the porcine iPSC extracts. The Chariot^TM reagent system was used to deliver the iPSC extracts into cultured porcine ear skin fibroblasts. The iPSC extracts-treated cells (iPSC-treated cells) were cultured for 3 days and used for analyzing histone modification and somatic cell nuclear transfer. Compared to the results for nontreated cells, the trimethylation status of histone H3 lysine residue 9 (H3K9) in the iPSC-treated cells significantly decreased. The expression of Jmjd2b, the H3K9 trimethylation-specific demethylase gene, significantly increased in the iPSC-treated cells; conversely, the expression of the proapoptotic genes, Bax and p53, significantly decreased. When the iPSC-treated cells were transferred into enucleated porcine oocytes, no differences were observed in blastocyst development and total cell number in blastocysts compared with the results for control cells. However, H3K9 trimethylation of pronuclear-stage-cloned embryos significantly decreased in the iPSC-treated cells. Additionally, Bax and p53 gene expression in the blastocysts was significantly lower in iPSC-treated cells than in control cells. To our knowledge, this study is the first to show that an extracts of porcine iPSCs can affect histone modification and gene expression in porcine ear skin fibroblasts and cloned embryos.     

仔猪黄痢K88-K99-987p-F41多价菌毛疫苗的制备及小鼠免疫试验

为了更好地预防仔猪黄痢,选取野生分离菌株HN2001（K88ab）、HN2002（K88ac）、HN2003（K88ad）、HN2004（K99）、HN2005（987p）和HN2006（F41）培养后用低温磁力搅拌法提取菌毛,制成5批多价菌毛混合油乳剂灭活苗。试验结果表明,5批多价灭活疫苗对小鼠的平均保护率达95．0％,为进一步进行本体动物试验提供了有价值的参考资料。     

盐胁迫对碱茅幼苗叶片内源激素、NAD激酶及Ca2+-ATPase的效应

为了研究盐渍生境对盐生植物碱茅Puccinellia chinampoensis体内部分信号分子的影响,采用不同浓度NaCl溶液处理碱茅植株,对幼苗叶片中部分信号分子及相关蛋白测定和分析.结果发现,低盐下碱茅无胁迫反应,高盐胁迫下,碱茅脱落酸(ABA)含量升高,生长素(IAA)、赤霉素(GA)含量均下降,细胞分裂素(ZR)含量趋于稳定;碱茅NAD激酶(NADKase)活性升高,提高代谢调节能力;同时Ca2 -ATPase活性增加,是对盐胁迫引起Ca2 浓度升高而做出的胁迫反应.表明了碱茅通过胞间信号相对含量的变化,调节NADKase活性、Ca2 -ATPase活性,提高耐盐能力.     

施钾对草地早熟禾根部可溶性碳水化合物及叶片氮，磷和钾含量的影响

以草地早熟禾（Poapratensis）品种男爵（Baronie）为试验材料,研究了不同施钾处理对根部0～10cm可溶性碳水化合物和叶片氮,磷和钾含量的影响。结果表明：在单施钾肥的情况下,随着施钾量的增加,越冬前草地早熟禾根系可溶性碳水化合物的含量也显著提高（P〈0．05）,这可能有助于提高植株越冬率,促进返青生长,而在同时施有氮、磷的情况下,碳水化合物含量间的差异并不显著,草地早熟禾叶片氮、钾含量也随着施钾量的增加呈显著增加的趋势（P〈0．05）。通过碳水化合物及叶片氮,磷和钾含量的测定,以N5g／m^2＋P20s3．33g／m^2＋K2O 6．68g／m^2处理对草坪返青及分蘖最佳。     

Cardiovascular Responses to Glibenclamide During Endotoxaemia in the Pig

Vanelli, G., Hussain, S.N.A., Dimori, M. and Aguggini, G., 1996. Cardiovascular responses to glibenclamide during endotoxaemia in the pig. Veterinary Research Communications, 21(3), 187-200The effects of blockading the ATP-sensitive potassium channel (K+ATP channels) on endotoxin-induced vascular derangements was studied. Escherichia coli endotoxin was infused (20 µg/kg per h) intravenously for 180 min into anaesthetized, mechanically ventilated, indomethacin-treated pigs. After 150 min of endotoxaemia, glibenclamide (a K+ATP channel blocker) was infused intravenously at 2 mg/kg per min for 5 min. The cardiovascular parameters were recorded before (control), every 30 min up to 150 min during endotoxaemia, and then at 5, 15 and 30 min after administration of glibenclamide. Infusion of endotoxin reduced the systemic arterial pressure to 60.6% ± 3.7% (p < 0.01) and increased the pulmonary arterial pressure by 75.9% ± 11.0% (p < 0.01) of the control values. Within 5 min, infusion of glibenclamide transiently but significantly reversed the systemic hypotension by raising the systemic vascular resistance, whereas the increased pulmonary arterial pressure was further augmented. Glibenclamide infusion did not influence the cardiac output. Within 30 min, the cardiovascular parameters had returned to the values induced by endotoxin, except for the systemic vascular resistance. Infusion of glibenclamide into normal pigs did not change the systemic pressure or resistance, but the pulmonary pressure and resistance were augmented transiently. These data suggest that, in pigs, the ATP-sensitive K+ channels may be one factor playing a role in the vascular changes due to endotoxaemia, especially in the systemic circulation.     

Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves

Escherichia coli (E. coli) strains were collected from young diarrheic calves in farms and field. Strains that expressed the K99 (F5) antigen were identified by agglutination tests using reference antibodies to K99 antigen and electron microscopy. The K99 antigen from a selected field strain (SAR-14) was heat-extracted and fractionated on a Sepharose CL-4B column. Further purification was carried out by sodium deoxycholate treatment and/or ion-exchange chromatography. Monoclonal antibodies to purified K99 antigen were produced by the hybridoma technique, and a specific clone, NEK99-5.6.12, was selected for propagation in tissue culture. The antibodies, thus obtained, were affinity-purified, characterized and coated onto Giemsa-stained Cowan-I strain of Staphylococcus aureus (S. aureus). The antibody-coated S. aureus were used in a co-agglutination test to detect K99+ E. coli isolated from feces of diarrheic calves. The specificity of the test was validated against reference monoclonal antibodies used in co-agglutination tests, as well as in ELISA. Specificity of the monoclonal antibodies was also tested against various Gram negative bacteria. The developed antibodies specifically detected purified K99 antigen in immunoblots, as well as K99+ E. coli in ELISA and co-agglutination tests. The co-agglutination test was specific and convenient for large-scale screening of K99+ E. coli isolates.     

A free-choice intake of mineral blocks in beef cows during the grazing season and in winter

A herd of 25 beef cows were offered four types of mineral blocks (A, B, C, D) with a different Ca to P ratio (A 0.6:1, B 0.8:1, C 2:1 and D 3.6:1). The Mg content was identical in all blocks whereas the Na content was equal in B, C and D, while the Na content in A block was double. The mineral block intake was monitored for 1 year, which was broken down to the grazing period (Pa) and the winter period (Wi). The intake of pasture grass and winter forage was assessed and the feed chemical composition was analysed; based on the obtained values, a macroelement balance (for Ca, P, Mg, Na and K) was calculated.The results showed the forage diet met requirements for all the observed macroelements (except in Na). The macroelement intake from forage was considerably higher than from mineral blocks. There were significant seasonal (Pa versus Wi) differences in B and C blocks intake. The consumption of D block was relatively invariable all year round; block A was refused in both seasons though it contained the highest amount of highly deficient Na.It can be concluded that mineral content in forage meets beef cattle requirements for Ca, Mg and K, P content is just satisfactory, and Na content is insufficient. It appears that cows control, to some extent, the Ca to P ratio in their diet by choosing the appropriate type of mineral block. However, the preference or refusal of some types of mineral blocks regardless of the season suggests the influence of other factors affecting mineral block intake which were not the subject of our observation.