东亚钳蝎毒生物提取物对人肝癌细胞体外增殖及凋亡的影响

目的：探讨东亚钳蝎毒生物提取物（PBMK）对人肝癌细胞的体外增殖及凋亡影响。方法：用MTT法测定PBMK对人肝癌细胞的体外增殖，用Tunel法测定其对人肝癌细胞的凋亡影响。结果：PBMK对人肝癌细胞体外生长有明显抑制作用，可高度诱导肝癌细胞的凋亡。经PBMK作用的癌细胞在相差显微镜下表现为细胞稀疏、收缩，胞质拉长。倒置显微镜下可见明显的细胞内空泡和细胞出泡现象。结论：PBMK可作为一种有效的癌细胞毒药物，具有开发应用前景。     

Reduction and reoxidation of the neurotoxin II from the scorpion Androctonus australis Hector

Reoxidation of the totally reduced scorpion neurotoxin II from Androctonus australis Hector (four disulfide bridges) has been investigated. The totally reduced toxin was highly insoluble in neutral and alkaline conditions, which prevented the use of the usual air oxidation process for renaturation. We tested a new method in which the reduced molecules were first solubilized in 10% (v/v) acetic acid and then oxidized by air through dialysis against a series of buffers with a slow pH gradient from 2.2 to 7.0 or 8.0. In this system, up to 95% of the protein was recovered in solution. Addition of reduced and oxidized glutathione accelerated refolding and also permitted a better recovery of fully active peptide as measured by both toxicity to mice and ability to displace ¹²⁵I radiolabeled toxin II from its binding site on rat brain synaptosomal fractions. The reoxidation reaction could also be monitored directly by high pressure liquid chromatography. A strong effect of guanidine hydrochloride concentration as well as of temperature was observed both on the solubility of the reoxidation intermediates and on the refolding pathway. Finally, the method used, i.e. dialysis reoxidation with a pH gradient from 2.2 to 8.0 in 0.1 M sodium phosphate, 0.1 M sodium chloride, 20 mM guanidine hydrochloride, 1 mM oxidized and reduced glutathione allowed regeneration in high yield (70%) of a reoxidized toxin form indistinguishable from the native toxin. A minor stable and inactive molecular species (about 30%) showing a difference in mobility by electrophoresis was also detected.     

Anaphylaxis to scorpion antivenin and its management following envenomation by Indian red scorpion,Mesobuthus tamulus

Mesobuthus tamulus is an Indian red scorpion that is responsible for numerous cases of scorpion stings in the Indian subcontinent. Antivenin, vasodilators, and benzodiazepines are medications of choice in the treatment of scorpion bites. Adverse reactions such as anaphylaxis to antivenin have been infrequently described in the literature. We, herein, present a case of a 42-year-old man stung by Indian red scorpion while gardening at home in India, who presented with extreme pain at the sting site and signs of cardio-toxicity. He was treated with scorpion antivenin and vasodilators but developed anaphylaxis to antivenin. We discuss management strategies. Anaphylaxis to antivenin should be on the differential during management of scorpion bites because classical signs of anaphylaxis may be absent.     

Human T-cell Kv1.3 potassium channel blockers: new strategies for immunosuppression

Blockers of the Kv1.3 channel, a voltage-gated K⁺ channel found in human T-cells, have recently been recognised as a general strategy for immunosuppression. These blockers are divided into two classes – peptidyl and non-peptidyl Kv1.3 channel blockers. This article covers the patents filed for Kv1.3 channel blockers dating from 1996 to the first quarter of 2000.     

羚蝎胶囊对大鼠局部脑缺血再灌注模型脑组织ATP酶的影响

目的了解羚蝎胶囊对脑缺血再灌注大鼠脑组织能量代谢的影响.方法将大鼠分为假手术组、模型组、治疗组3组,用MCAO法复制大鼠局部脑缺血再灌注模型,观察各组大鼠神经功能缺损积分和脑组织ATP酶的变化.结果治疗组再灌注后3 h神经功能缺损积分显著低于模型组,治疗组Na -K -ATPcase、Ca2-ATPcase、Mg2 -ATPcase活性均较模型组高(P＜0.01或P＜0.05).结论羚蝎胶囊能提高大鼠局部脑缺血再灌注模型脑组织ATP酶的活性.     

Revealing the Function and the Structural Model of Ts4: Insights into the “Non-Toxic” Toxin from Tityus serrulatus Venom

The toxin, previously described as a “non-toxic” toxin, was isolated from the scorpion venom of Tityus serrulatus (Ts), responsible for the most severe and the highest number of accidents in Brazil. In this study, the subtype specificity and selectivity of Ts4 was investigated using six mammalian Nav channels (Nav1.2→Nav1.6 and Nav1.8) and two insect Nav channels (DmNav1 and BgNav). The electrophysiological assays showed that Ts4 specifically inhibited the fast inactivation of Nav1.6 channels, the most abundant sodium channel expressed in the adult central nervous system, and can no longer be classified as a “non-toxic peptide”. Based on the results, we could classify the Ts4 as a classical α-toxin. The Ts4 3D-structural model was built based on the solved X-ray Ts1 3D-structure, the major toxin from Ts venom with which it shares high sequence identity (65.57%). The Ts4 model revealed a flattened triangular shape constituted by three-stranded antiparallel β-sheet and one α-helix stabilized by four disulfide bonds. The absence of a Lys in the first amino acid residue of the N-terminal of Ts4 is probably the main responsible for its low toxicity. Other key amino acid residues important to the toxicity of α- and β-toxins are discussed here.     

蝎毒多肽有效组分对辐射后M-NFS-60细胞的促增殖作用

目的：观察蝎毒多肽（SVP）及IL-3对M—CSF依赖细胞株M—NFS-60促增殖作用以及对辐射后M—NFS-60细胞增殖的影响。方法：采用Alamar Blue摄入法检测细胞的增殖情况。①选择10个细胞浓度．分别于0h、2h、4h、5．5h和20h,检测细胞的增殖情况,确定M—NFS-60细胞对Alamar Blue反应所需的最佳细胞密度和孵育时间;②采用最佳细胞密度和孵育时间,观察10个SVP组分促进M—NFS-60细胞的增殖作用,筛选出SVP的有效组分;⑧M—NFS-60细胞经60Coγ-射线照射后分别给予生理盐水、SVP、IL-3、SVP＋IL-3处理48h,测定细胞增殖情况。结果：①M—NFS-60细胞的接种密度为5×10^4cells／mL时,对AlamarBlue的还原能力最强,孵育时间8h后,M—NFS-60细胞对Alamar Blue的还原率开始增强,72h达最高;②从10个SVP组分中筛选出Ⅱ3、Ⅳ能明显促进M—NFS-60细胞的增殖;③SVPⅡ3、Ⅲ3、Ⅳ或IL-3处理辐射后的细胞48h,细胞的增殖率（％）分别为121．58±2．62（113）、123．39±4．45（Ⅲ3）、123．51±5．04（Ⅳ）、140．12±1．68（IL-3）,与空白对照组（100．00±0．01）相比差异有显著性（P〈0．01）;113、Ⅲ3、Ⅳ分别与IL-3联用处理细胞48h后的增殖率分别为163．98±9．20（113±IL3）、159．89±8．31（Ⅳ-IL3）、148．92±9．74（Ⅲ3±IL3）,与SVP处理组比较差异有显著性（P〈0．05）。结论：蝎毒中存在促进M—NFS-60增殖的有效多肽组分,这些多肽组分能明显促进辐射后M—NFS-60细胞的增殖,并与IL-3对辐射后M—NFS-60细胞的增殖具有协同作用。     

Management of scorpion sting in Morocco

Scorpion stings are the first cause of poisoning, and represent between 30 and 50% of all cases reported to the Moroccan Poison Control Centre. Concerned by the size of the problem, we have paid special attention to this pathology. Through retrospective and prospective studies, it has been possible to determine the nature and the chronology of clinical stages, as well as the epidemiological, clinical and therapeutic factors of gravity. On this basis, we worked out a new management to provide support for patients. This management will standardize support provided at the national level, and will reduce the number of lethal case and rationalize spending by reviewing medication, transfer of patients and hospital care. This standardization is an essential component of the national strategy against scorpion poisoning. Other components include training of medical staff, awareness campaigns, and information systems to monitor lethal cases. A survey over five years shows a reduction in the number of lethal cases and rationalization of costs. Medical care provided rests upon the distinction between patients stung by scorpions and patients actually poisoned. The first category of patients will be monitored up to four hours after the scorpion sting, while poisoned patients will be transferred to an intensive care unit.     

蝎毒多肽提取物对白血病NOD/SCID小鼠MMP2、MMP9表达的影响

目的:观察蝎毒多肽提取物(PESV)对白血病NOD/SCID小鼠基质金属蛋白酶(MMP)2、MMP9表达的影响,探讨PESV对白血病细胞外基质降解和髓外浸润的干预机制.方法:首先选取急性白血病患者骨髓单个核细胞注入经过铯-137源照射的NOD/SCID小鼠体内,建立人白血病NOD/SCID小鼠髓外浸润模型;再将实验小鼠随机分组,Ⅰ组、Ⅱ组、Ⅲ组分别注射不同浓度的PESV,Ⅳ组为模型组注射生理盐水,另设Ⅴ组为空白对照组,观察各组小鼠外周白细胞计数、血涂片及生存状态;观察4周后处死小鼠,用Real time PCR方法检测小鼠体内MMP2、MMP9表达水平.结果:给药各组小鼠MMP2、MMP9表达水平均明显低于模型组(P < 0.05),且各组间表达水平随给药浓度增加而减低.给药各组小鼠外周血白细胞计数、血涂片及生存状态也均优于模型组.结论:PESV可以抑制白血病NOD/SCID小鼠体内MMP2、MMP9过度表达,对白血病细胞外基质降解和髓外浸润具有明显阻抑作用.