蝮蛇毒抗凝血活酶组份及几种蛇毒粗毒对人红细胞和血小板的作用

蝮蛇毒抗凝血活酶组分及蝗蛇毒、圆斑蝰蛇毒和眼镜蛇毒粗毒,不仅能够水解血浆中的磷脂,而且还能水解完整人红细胞膜和完整人血小板膜上的磷脂。但是五步蛇毒和金环蛇毒粗毒却不能水解完整人血小板膜上的磷脂。 扫描电镜观察表明,由于抗凝血活酶组份和几种蛇毒粗毒的作用,人红细胞和人血小板的形态发生了巨大的变化;人红细胞由正常的双圆盘形变成带刺的小球,人血小板的外形变成蜂窝状。     

蝮蛇抗栓酶治疗小儿脑炎致急性偏瘫血中6—K—PGF1α,TXB2的变化

应用蝮蛇抗栓酶治疗小儿脑炎致急性偏瘫共25例。沿疗前后检测血中的6-K-PGF_1x及 TXB_2.结果表明:治疗后血中 6-酮-前列腺素 F_1x 明显增高,血栓素 B_2明显降低。推测蝮蛇抗栓酶之所以具有去纤、溶栓、扩血管等作用,可能与血中前列腺素、血栓素 B_2水平变化有关。     

Crystal structure determination of basic phospholipase A2 from venom of Agkistrodon halys pallas by molecular replacement method

Basic phospholipase A₂ (BPLA₂) from the venom of Agkistrodon halys pallas has a strong ability to hemolyze erythrocytes. The asymmetrical unit of P2₁2₁2₁ crystal of BPLA₂ contains two molecules. Self-rotation function was used to study the orientation relationship of these two molecules. Cross-rotation and translation functions were then used to determine the orientations and positions of the two molecules in the unit cell. The model building and preliminary structure refinement were carried out. The result shows that the two molecules in the asymmetrical unit of orthorhombic crystal are related by a non-crystallographic 2-fold symmetry axis.     

蝮蛇毒碱性磷脂酶A_2基因的克隆

从蝮蛇毒腺中抽提总ＲＮＡ．利用人工合成寡核苷酸引物作逆转录,以ｃＤＮＡ为模板进行体外扩增,获得磷脂酶Ａ２（简称ＰＬＡ２）基因,克隆至ｐＢＳ－ｋｓ载体中。通过对３个碱性ＰＬＡ２（简称ＢＰＬＡ２）基因单独克隆分别作ＤＮＡ全序列分析,推导ｐｒｏ－ＢＰＬＡ２由１３８个氨基酸残基构成,与已测定的部分氨基酸序列比较,基本相符。该基因成功的克隆,不仅推导出ＢＰＬＡ２的蛋白质全序列,也为进一步开展蛇毒功能肽蛋白质工程的研究工作打下了良好的基础。     

皖南尖吻蝮蛇毒出血毒素Ⅳ的纯化与初步鉴定

从皖南尖吻蝮蛇（Ａｇｋｉｓｔｒｏｄｏｎａｃｕｔｕｓ）毒液中经ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ和ＳｅｐｈａｃｒｙｌＳ－２００两步凝胶柱层析首次纯化出一种中分子量出血毒素（简称ＡａＨⅣ）．经ＳＤＳ－ＰＡＧＥ和等电聚焦凝胶电泳测定其分子量为４４ｋＤ,等电点为ｐＨ５．０．从５００ｍｇ粗毒中可获得２０ｍｇＡａＨⅣ纯品．ＡａＨⅣ有较强的出血活性,最小出血剂量（ＭＨＤ）为０．４μｇ．     

五步蛇主要数量性状与排毒量的相关分析

本文首次报道了五步蛇身体主要部位的数量性状与其排毒量的关系。其中我们发现与排毒量有关的部位是头积、体粗、体重、佛指长。     

尖吻蝮咬伤与心脏损害：附20例分析

本文报告尖吻蝮咬伤并心脏受累20例。经中西医结合治愈19例(95％),死亡1例(5％),并对其发病机理及治疗进行了讨论。     

濒危植物秦岭石蝴蝶的SCoT遗传多样性分析

利用筛选到的24条SCoT引物,分析秦岭石蝴蝶人工繁育和野生种群60份材料的遗传多样性和遗传结构,为秦岭石蝴蝶的濒危机制和制定种群保护与修复策略提供理论依据。结果显示:(1)60份秦岭石蝴蝶供试样品的观察等位基因数平均为1.51,有效等位基因数平均为1.31,Nei's基因多样性平均为0.2305,Shannon指数平均为0.3703,表明供试材料种群的遗传多样性较低。(2)种群间的遗传距离与遗传相似系数分析结果表明,3个种群60份供试材料间的遗传相似系数为0.9551~0.9705,平均相似系数0.9634,进一步表明供试材料之间的遗传相似性极高,遗传背景较为狭窄。(3)秦岭石蝴蝶种群分子方差分析结果显示15%的变异来自于种群间,而85%的变异来自于种群内部。综合分析表明,秦岭石蝴蝶各种群内和种群之间的遗传多样性较低,遗传背景狭窄,这可能是秦岭石蝴蝶适应环境能力差,从而导致其濒危的重要原因之一。     

长白蝮蛇类凝血酶基因的克隆及分析

从长白蝮蛇（Agkistrodon halys Ussuriensis)毒腺中抽提总RNA,采用RT－PCR扩增其类凝血酶基因,经全序列测定,类凝血酶基因Ussurin全长为708个核苷酸,即编码236个氨基酸;根据同源性,推测它的活性中心为His^43,Asp^88和Ser^182;二硫键为Cys^7-Cys^141,Cys^28-Cys^44,Cys^76-Cys^234,Cys^120-Cys^188,Cys^152-Cys^167和Cys^178-Cys^203。该蛇毒类凝血酶cDNA序列及推导的氨基酸序列均为首次报道。     

Terbium(III) fluorescence probe studies on metal ion-binding sites in anticoagulation factor I from Agkistrodon acutus venom

Anticoagulation factor I (ACF I) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X-binding protein with marked anticoagulant activity. Present studies show that holo-ACF I assumes a compactly folded structure in the range of pH 5–6, in which the most interior Trp residues and quenchers are adjacent. Tb³⁺ ions can completely replace both Ca²⁺ ions in holo-ACF I, as determined by equilibrium dialysis. Although the two Tb³⁺ ions in Tb³⁺-ACF I have slightly different luminescence efficiencies, both have similar quenching effects on the intrinsic fluorescence, suggesting that probably there are same numbers of Trp residues close to both Tb³⁺-binding sites. Two Tb³⁺-binding sites with similar apparent Tb³⁺ association constant values, (1.69 ± 0.02) × 10⁷ M^–1 and (1.42 ± 0.01) × 10⁷ M^–1, respectively, were further identified through Tb³⁺ fluorescence titration. In addition, it has been confirmed from the titration of holo-ACF I and Tb³⁺-ACF I with NBS that only interior Trp residues are involved in the energy transfer to Tb³⁺ ions and that all accessible Trp residues located in the surface of holo-ACF I have similar affinity to NBS, while those located in the surface of Tb³⁺-ACF I have two different kinds of affinity to NBS, which strongly suggests a conformational change of holo-ACF I upon substitution of Tb³⁺ for Ca²⁺. The results show that although the Tb³⁺-altered conformation of ACF I cannot support the binding of Tb³⁺-ACF I with FXa, determined by nondenaturing PAGE, Tb³⁺ ions are effective and useful fluorescence probes to analyze the structures and properties of Ca²⁺-binding sites in ACF I.