Effects of three methionine sources in diets on temporal B0AT1 and ASCT2 mRNA expression in different intestinal segments of cobia,Rachycentron canadum

A 4‐week feeding trial was conducted using five groups of hybrid grouper (61.15 ± 0.15 g) to explore the potential effects of three methionine (Met) sources. Five isonitrogenous and isolipidic diets were formulated that included a fishmeal (FM) diet; nonsupplemented (NS) diet; or NS diets with the addition of the L‐methionine (L‐Met), DL‐methionine (DL‐Met), or coated Met (Co‐Met) to obtain the same Met level as the FM diet. Fish were randomly distributed into sea cages (30 fish per cage). Weight gain and specific growth rate in the DL‐Met group and FM group were significantly higher than those in all other groups (p < .05). In the proximal and distal intestines, of the 20 gene and time combinations (10 per gene), there were 14 combinations (70%) in which there were no significant differences in gene expression levels between the FM and DL‐Met groups (p > .05). The main reason for the same growth effect between the DL‐Met and FM groups may be attributed to the synchronized absorption at most time points after feeding, which had similar expression patterns of B⁰AT1 and ASCT2 of the proximal and distal intestines between two groups.     

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.     

菱传粉生物学研究

以6 个菱品种为试材，对花器形态特征、群体花期及单花开花动态、开花时间、柱头可授性、花粉活力、杂交指数 及传粉形式等进行观察测定。结果表明：菱花单生，两性，单花花期15 h，群体花期在5～10 月，盛花期在7～8 月；不同 品种在一天中的开花高峰期存在差异；柱头可授期主要在萼片开裂后2 h 内，结实率在50% 以上；花粉在自然授粉条件下活 力可保持7.5 h；以自花授粉为主，同时存在一定程度的无融合生殖和少量异花授粉。     

蓖麻环核苷酸门控离子通道RcCNGC2克隆与盐胁迫下表达分析

为探讨RcCNGC2在蓖麻耐盐中的作用,以通蓖5号叶片为材料,克隆获得环核苷酸门控离子通道RcCNGC2完整编码区序列(CDS),并对该序列进行序列比对、蛋白结构分析,构建多元表达载体,分析了在盐胁迫下蓖麻RcCNGC2基因根、茎、叶组织中6个时间点的表达量变化。结果显示,RcCNGC2 CDS长度为2 148 bp,编码715个氨基酸,蛋白分子量为34.15 ku,等电点为9.96,存在5个跨膜结构域。氨基酸序列一致性分析表明,RcCNGC2与橡胶树一致性最高,达89.15%。qRT-PCR分析结果显示,NaCl处理后RcCNGC2表达量在根中上调且差异显著,在茎、叶中随NaCl处理时间延长而显著减少。综上,RcCNGC2在蓖麻受到盐胁迫时起重要信号传导作用。     

雪山草鸡感染传染性法氏囊病病毒新型变异株的鉴定

传染性法氏囊病病毒(infectious bursal disease virus,IBDV)新型变异株正在我国广泛蔓延,给商品肉鸡群和蛋鸡群带来了新的威胁。本研究对一个疑似发生非典型传染性法氏囊病的地方品种雪山草鸡群进行了RT-PCR检测,并对分离的3株毒株进行了基因测序及序列分析。结果显示,引起该鸡群发病的病原是IBDV新型变异株,属于A2dB1基因型;这3个毒株的VP2高变区编码蛋白上含有IBDV变异株的特征性氨基酸,也具有IBDV新型变异株的独特氨基酸。结果提示,我国地方品种鸡群中也有IBDV新型变异株的流行,现有的部分商品化IBDV疫苗已不能有效预防IBDV新型变异株的流行。     

一种高质量天门冬基因组DNA提取方法

针对天门冬[Asparagus cochinchinensis(Lour.)Merr.]成熟叶片富含多糖、多酚、蛋白质及其他次生代谢物质的特点,采用改良CTAB法对其基因组总DNA进行提取,并对DNA进行质量检测。结果表明,改良CTAB法可从天门冬成熟叶片中获得纯度高、完整性好的总DNA,其OD260 nm/OD280 nm为1.86,OD260 nm/OD230 nm为2.36,浓度为390μg/mL,产率达11.7μg/g,多糖、多酚和RNA杂质被去除干净,无降解现象,说明该方法提取的天门冬基因组总DNA质量较高。     

水生动物肌醇营养研究进展

肌醇是大多数水生动物保持健康、促进生长发育必需的营养素,它在细胞膜磷脂合成、神经递质传递和肝脏脂质转运等方面发挥重要作用。饲料中肌醇缺乏导致水生动物生长不良、体表充血、鳍条糜烂、体色发黑、脂肪肝等症状。本文综述了肌醇的理化性质,肌醇在水生动物营养中营养生理作用、需要量及影响因素等,同时还介绍了肌醇的来源和测定方法,并展望了肌醇在水生动物营养中进一步的研究和应用方向。