C端序列的删除对非出血重组蛇毒纤溶酶的影响

[目的]研究C端序列对非出血重组纤溶酶(rFⅡ)特异性、活性和热敏感性的影响.[方法]通过SOEing PCR方法构建删除C端序列的突变体,突变体和rFⅡ分别在P.pastoris中诱导表达.表达和纯化的蛋白用SDS-PAGE和Westeren blot进行鉴定.之后分析其生化特性.[结果]rFⅡ及突变体通过SDS-PAGE和Westeren blot得到证实.生化分析揭示:①突变体对显色底物N-(p-Tosyl)-Gly-Pro-Lys-pNA的催化效率(Kcat/Km)是rFⅡ的1.9倍;②突变体和rFⅡ对氧化的胰岛素B链拥有共同的优先裂解位点,可是在随后的裂解中开始展现差异;③突变体显示了更高的纤(原)活性;④热处理表明突变体相较r FⅡ对温度的增加更敏感;⑤通过圆二色谱测量,突变体的螺旋比例有所增加.[结论]删除的序列参与rFⅡ的特异性、活性和热敏感性,该序列可作为下一步优化的候选序列.     

一种新的五步蛇蛇毒类凝血酶Cdna的克隆和序列分析

目的：克隆出五步蛇的类凝血酶cDNA，为研究其结构和功能的关系。并为下一步基因表达开发抗血栓药物提供依据和基础。方法：从五步蛇蛇腺中提取了总RNA，通过反转录合成第一链cDNA，经PCR扩增出五步蛇毒腺中类凝血酶TLE1的cDNA片段，并将其连接到质粒载体扩增，然后进行DNA测序。结果：成功克隆出一种新的五步蛇类凝血酶的cDNA片段。此片段全长794bp，包括编码部分的全长为777bp。推导其编码的蛋白质序列为258个氨基酸，其中包括18个氨基酸的信号肽，6个氨基权的前肽和234个氨基酸的成熟氨基酸顺序。TLE1成熟肽与其它蛇毒类凝血酶相比，蛋白质一级结构具有较高的同源性。结论：从五步蛇中成功克隆出一 种新的类凝血酶cDNA，并确定了它的DNA顺序。     

带蒂胸腹轴型皮瓣早期修复上肢深度蛇伤溃疡

总结胸腹轴型皮瓣早期修复上肢不同部位的深度蛇伤溃疡的经验，评价其应用价值。本组１６例，胸脐皮瓣２例，侧胸腹皮瓣１例，髂腰皮瓣５例，下腹皮瓣６例，下腹分叶皮瓣１例，髂腰加下腹Ｙ形皮瓣１例。术后１例皮瓣远端坏死，３例轻度感染，１６例断蒂后全部成活。随访３个月～７年，皮瓣质地和弹性均好，手功能基本恢复。认为，带蒂胸腹轴型皮瓣是修复上肢深度蛇伤溃疡理想的方法。     

蝮蛇抗栓酶-3治疗糖尿病并发症94例

应用腹蛇抗栓酶-3治疗94例糖尿病患者并发的脑血栓形成、高脂血症、动脉硬化、神经病变和心绞痛取得较好的疗效。脑血栓形成急性期采用冲击疗法,瘫痪肢体功能恢复迅速、合并神经病变者,治疗后症状消失、神经功能改善。TG、TC、β-脂蛋白水平和TC/HDL比值均降低(P<0.01),HDL水平上升(P<0.01),对比治疗前后血6-Keto-PGF_(1α)、TXB_2和纤维结合素FN的水平改善,具有统计学意义(P<0.01)。     

Cross-neutralisation of Australian brown and tiger snake venoms with commercial antivenoms: Cross-reactivity or antivenom mixtures?

Recently it has been suggested that the Australian snake antivenoms made by CSL Ltd. are in fact not truly monovalent and may contain antibodies to other snake venoms because the horses are injected with multiple snake venoms. It is unclear to what extent various monovalent antivenoms can neutralise the effect of other venoms, whether this is due to a mixture of antibodies or true cross-reactivity, and whether this has any clinical significance. We aimed to study the immunological and functional properties of brown snake (Pseudonaja spp.) antivenom (BSAV) and tiger snake (Notechis spp.) antivenom (TSAV) against their respective venoms using enzyme immunoassays (EIA) and in vitro clotting studies. There was significant overlap between the two antivenoms with both TSAV and BSAV being detected by EIA on brown snake venom (BSV)-coated and tiger snake venom (TSV)-coated wells, respectively. In a competition EIA, increasing amounts of immunoaffinity-purified hen anti-brown antibodies (IgYp) mixed with TSAV reduced TSAV measured on TSV-coated wells. Both BSAV and TSAV prevented the clotting activity of both venoms. IgYp also prevented the clotting activity of TSV, suggesting true cross-reactivity. The cross-reactivity of TSAV and BSAV with BSV and TSV, respectively, was likely due to each being a mixture of anti-brown and anti-tiger antibodies, but there was partial cross-reactivity demonstrated by the effect of IgYp. Single-polyvalent antivenom for brown snake and tiger snake may be feasible in the future.     

7种蛇类药材的等电点鉴别研究

用改进的蛋白质等电点法对7种蛇类药材进行了鉴别研究。结果表明不同种类的蛇,等电点有所不同。重现性好(CV<0.5),其等电点均值的差别(P<0.01)有非常显著的意义,可以作为去皮或粉末状蛇类药材鉴别的依据之一。     

Two Sri Lankan cases of identified sea snake bites, without envenoming.

Sea snakes are among the most venomous creatures encountered around coasts and reefs, in estuaries, rivers and at sea. Their venoms are more toxic than those of land snakes. However, they are rarely aggressive or menacing. Bites have become unusual with the advent of modern fishing methods but the two encounters we report, in the Indian Ocean off the shores of Sri Lanka, emphasise that sea snake bites may not result in envenoming.     

蛇毒抗栓酶对氧自由基致血小板变化的影响

次黄嘌呤／黄嘌呤氧化酶体系产生的氧自由基在体外能明显增加血中板膜微粘度，脂质过氧化代射产物丙二醛含量，血小板胞内游离钙浓度和血小板５－羟色胺的释放量。０．０３ｕ／ｍｌ的蛇毒抗栓酶明显抑制前二种效应，但促进血小板释放５－羟色胺，增加血小板胞内游离钙浓度，结果表明蛇毒抗栓酶对血小板膜有保护作用。     

混编或空载货车通过中等跨度桥梁横向振动机理分析

通过对现场实测桥梁(中等跨度上承式及半穿式钢桁梁)振动波形的分析,探讨了混编列车通过桥梁时易发生脱轨事故的原因。提出当车速为60～70km/h时,带有磨耗型踏面的空载货车横向蛇行振动频率为1．5～2．9Hz,与桥梁带载横向自振频率相符,引起共振。