新标准“注射用克痛宁”临床应用镇痛效果观察

目的　评估按国家药品生物制品检定所修订的国家标准生产的“注射用克痛宁” (电泳纯和免疫纯 )的临床应用效果及毒副作用。方法　各种中、重度急慢性疼痛患者 92例 ,肌肉注射给药 ,70 μg/次 ,70～ 1 40 μg/天。慢性疼痛病例一般按疗程用药 ,每疗程 5天 ,急性疼痛病例及部分慢性疼痛病例疼痛消失即停药。以病人曾用过的镇痛药疗效作为历史对照。结果　总有效率 96 .7% ,总显效率 82 .6 % ,首支总有效率 93.5 %。对癌痛组的首支有效率稍低 ,为 84 .2 %。一般在肌注后 30～ 6 0 min起效 ;对急性疼痛疗效可持续 6～ 1 0 h,对慢性疼痛大多可持续 1 2 h或 2 4 h以上。对阿片类药物成瘾患者也有良好效果。少数病人出现轻度口干、恶心、头晕等不良反应。结论　新标准“注射用克痛宁”对各种急慢性疼痛均有良好的镇痛效果 ,起效快、镇痛作用强而持久、不成瘾、毒副作用小 ,且具有潜在的戒毒作用。优于原地方标准的“克痛宁”。     

新克痛宁术后镇痛效果观察

目的了解眼镜蛇毒制剂新克痛宁对术后镇痛效果。方法对７２例胫腓骨折病人,在连续硬膜外阻滞麻醉下行切开复位内固定术,术后随机分为４组（每组１８例）向硬膜外腔注药：Ａ组新克痛宁０．２５ＩＵ／ｋｇ;Ｂ组新克痛宁０．１２５ＩＵ／ｋｇ加０．５％利多卡因液１０ｍｌ;Ｃ组吗啡２ｍｇ;Ｄ组吗啡１ｍｇ加０．５％利多卡因液１０ｍｌ。结果镇痛持续时间Ａ、Ｂ组明显延长,Ａ与Ｂ组分别为（４１２±２８）ｍｉｎ,与Ｃ、Ｄ组比较差异显著（Ｐ＜０．０５）。注药后３０～１２０ｍｉｎ,患肢足背皮肤温度,Ａ、Ｂ组亦高于Ｃ、Ｄ组（Ｐ＜０．０５）,且无不良反应。结论新克痛宁术后镇痛效果比吗啡好。     

新标准“注射用克痛宁”长期毒性试验

目的 评估新标准克痛宁的长期毒性。方法 以较高剂量新标准克痛宁连续４５天给Ｂｅａｇｌｅ犬注射,检测用药期及停药后和各项生理学指标,并对脏器进行病理学检验。结果 高低剂量组与对照组的各项检测指标均地明显差异。结论 新标准克育宁对狗无长期毒性。     

给兔的蛛网膜下腔注射蛇毒新克痛宁的实验观察

观察不同剂量新克痛宁注入兔蛛网膜下腔后反应及对神经组织作用.结果表明:新克痛宁0.3～1.2μkg~(-1)无不良反应.光镜下多极神经元胞体、核、核仁及尼氏体的分布、形态无异常,电镜下见细胞膜、细胞核、核仁及线粒体无异常改变.实验结果证实,新克痛宁对神经组织均未引起组织细胞学方面的变化.     

克痛宁配合中药治疗坐骨神经痛182例分析

克痛宁是从眼镜蛇毒中提纯的神经毒素制剂,我科１９９０～１９９６年应用克痛宁配合中药治疗坐骨神经痛１８２例,疗效满意,现报告如下。１临床资料１．１一般资料（１）年龄与性别：男８２例,女１００例,３０岁以下４２例,３１～５０岁１０２例,５１岁以上３８例。...     

眼镜蛇毒研究与临床应用近况

用眼镜蛇整体泡酒饮服,治疗腰腿痛,祖国医学已有记载［１］,已有悠久历史。３０年代国外应用眼镜蛇毒治疗关节痛、神经痛,临床应用数百例,后因起效慢等原因,而终止研究与应用。我院于６０年代初开始对眼镜蛇毒进行系统研究,如原料标准,注射剂质控标准、安全试验、半数致死量（ＬＤ５０）试验、药理学、药效学、长毒以及“三致”试验等。结果表明：蛇毒及其制剂,治疗风湿痛、神经痛等疗效高于毒副反应。同时我们又研究以眼镜蛇毒为君药与中药伍用,治疗肝癌及消化道癌,亦收到了较好的疗效。这对深入研究与开发蛇毒新药是大有前途的     

眼镜蛇毒因子的临床应用

眼镜蛇毒因子（cobra venom factor,CVF）系眼镜蛇毒中的一种蛋白质。近年来,CVF的理化性质、分子生物学特性、免疫学作用以及基因序列等已逐渐被揭示[1,2]。随着对CVF的研究越来越深入,其研究价值越来越广泛,特别是在临床应用方面,正日益引起广大学者的重视。本文对CVF的临床应用现状作一简要综述。     

蛇毒抗凝剂的临床应用

利用蛇毒制成抗凝剂国外已有27年历史,国内对此项研究开始于1981年,并已广泛应用于临床。1963年以来,国外已有红口蝮蛇毒、矛头蝮蛇毒、东部菱斑响尾蛇毒、尖吻蝮蛇毒等商品供应于临床,国内已有蛇岛蝮蛇抗栓酶、长白山蝮蛇抗栓酶、江淅蝮蛇抗栓酶、去纤维蛋白酶等产品已相继在临床上用来治疗血栓、栓塞等闭塞性血管疾病。经基础研究和上千例次临床实践证明,在我国,蛇毒抗凝剂具有药源广;疗效佳、毒副作用较少的特点,部分产品已居国内外领先地位,是一种得推广应用的药物。本文简要介绍该药的临床应用,供用药时参考。     

临床应用分娩镇痛的研究进展

分娩镇痛有多种方法,一个半世纪以来学者们就不断地探讨着如何使产妇在清醒状态下分娩,无痛苦地诞生新的生命,但至今尚无一种绝对安全、简单、能普及的分娩镇痛方法。1992年美国妇产学院分娩镇痛委员会指出,理想的分娩镇痛必须具下列特征：对母婴影响小;易于给药,起效快,作用可靠,能满足整个产程的镇痛需要;避免运动阻滞,不影响宫缩及产妇运动;产妇清醒,可参与分娩过程;必要时可满足手术的需要。鉴于以上情况,分娩镇痛方法较难选择,本文就分娩镇痛相关理论及近年来国内外的一些研究进展作一综述。     

抗蛇毒血清临床应用体会

自1984年以来,我们应用抗蛇毒血清治疗各类毒蛇咬伤600余例,取得了满意的疗效.现就我们在临床应用中的一些体会介绍如下.1 运用抗蛇毒血清应及时足量抗蛇毒血清治疗毒蛇咬伤具有见效快,疗效确切等优点.但由于起初我们缺乏经验,病人就诊后,无论病情轻重,一律予先试敏后用药,结果一些如银环蛇咬伤等病情较危重者,试敏时间未到,或试敏阴性后在注药过程中,患者已出现呼吸麻痹等严重中毒症状,而被迫采取气管插管等急救措施,既给治疗与护理带来了许多困难,又影响疗效,更增加了病人的痛苦.另外,在及时用药的同时,把握好用药剂量,也极为重要.若药物用量不足,则全身中毒症状难以控制,或病情继续发展、恶化,甚者死亡.笔者曾遇1例尖吻蝮蛇咬伤严重中毒的患者,入院后即予注射精制抗五步蛇毒血清80001U,中毒症状未能得到控制,并进一步发展、加重,准备再予追加剂量时,患者拒绝而自动出院,结果死于多脏器功能损害.通过临床实践,吸取教训,我们总结出了“以缓治轻,以快治重”的给药方法和“酌情用量,中病方止”的用药原则,既病情较轻者,采用常规试敏阴性后用     

科博肽肠溶胶囊治疗晚期中、重度癌症疼痛疗效观察

目的 探讨科博肽肠溶胶囊对晚期癌症患者的止痛效果。方法 每6h口服科博肽肠溶胶囊280μg/次,常规剂量每天560μg,在用药过程中根据疼痛缓解程度调整剂量由280μg～420μg～560μg递增。结果 68例中、重度癌痛患者,显效41例,有效21例,无效6例,总有效率91．2％,药物最小日剂量280μg,最大日剂量为840μg,所有患者均可耐受,未见有药物依赖性。结论 科博肽肠溶胶囊作为控制中、重度癌痛的药物,其镇痛效果满意,副反应少,服用方便,安全,值得临床推广应用。     

Screening of plants containing Naja naja siamensis cobra venom inhibitory activity using modified ELISA technique

Enzyme-linked immunosorbent assay (ELISA) has been modified for screening plants with antagonistic activity to Naja naja siamensis cobra venom. Aqueous extracts from plants were investigated for their inhibitory effects on the binding of anti-cobra venom antibody to antigen, cobra venom, fixed onto 96-well microtiter plates. Ingredients in extracts were allowed to react with immobilized venom before the subsequent addition of antivenom antibody. Venom components affected by exposure to the extracts, unable to interact with their specific antibody, were predicted to be unable to bind to their native destinations or natural receptors. Curcuma cf. zedoaria, an old Thai medicinal plant, showed clear inhibitory activity in the ELISA test. Neurotoxin and protein degradative enzymes, major components in venom, were identified as targets of this extract in Western immunoblotting analysis. Ingredients in the extract showed high affinity to the toxin in competition assay by immunoprecipitation. The extract attenuated toxin activity by extending contraction time of diaphragm muscle after envenomation and had a potency to protect cellular proteins from venom degradative enzymes. Curcuma parviflora, with less activity in ELISA, exhibited acceptable results in two experiments but negative results in two experiments, whereas Curcuma longa, having low activity in the ELISA test, never showed any favorable results. Screening of 36 samples could classify plants into an inhibition range of 0 to 86%. This modified ELISA is recommended as a preliminary screening method for inhibitors with a large number of samples.     

A comparative in silico characterization of functional and physicochemical properties of 3FTx (three finger toxin) proteins from four venomous snakes

Snake venom is an abundant resource of diverse pharmacologically bioactive proteins and peptides and a good natural source of drug lead compounds and used as important research tools in the field of toxicology, pharmacology and neuroscience. Three finger toxins (3FTx) is an important super-family of snake venom proteins which has a conserved three finger like appearance in three dimensional structures. Members of 3FTx family show a wide array of pharmacological effects by targeting different receptors and ion channels with high specificity and many of them are being investigated as potential drug target. Therefore, with a vision to verdict a new edge and attempt we determined the amino acid compositional (%) profile, physiochemical properties, secondary structural and functional analysis and phylogenetic relationship of three finger toxins present in four different elapid snake species namely, Naja naja, Astrotia stokesii, Hydrophis cyanocintus and Pelamis platura using different bioinformatics tools. From the outcome of the current studies, it will be possible to know about a range of biological functions which are responsible mainly for the glowing amino acid composition profile of these proteins. Amino acid composition (%) profile although represents differential amount of different amino acid residues which encompasses a family precise model but all the protein sequence have a conserved amount of cysteine. The analysis of physicochemical properties can be used as a basic approach to contribute in developing rational drug through protein engineering and understanding different physiological function which will be beneficial for the welfare of human being.     

虎纹捕鸟蛛毒的生物学活性鉴定

本文报道了采集广西产虎纹捕鸟蛛（Selenocosmia huwena)毒液的方法,并对粗毒进行了部分生物学活性的测定,该粗毒对小鼠和蜚蠊的LD50分钟为1．16mg/kg和300ug/g,该粗毒具有透明质酸酶,碱性磷酸酶,蛋白水解酶和脱氧核糖核酸酶活性,未测到磷脂酶A和胆碱脂酶性,通过电生理实验发现该粗毒含有阻断蟾蜍神经肌肉接头传递的毒素,并观察到当小鼠接受腹腔注射致死剂量（5mg/kg)粗毒后便迅速出现呼吸麻痹。     

Classification of elapid snake neurotoxins and cytotoxins according to chain length: Evolutionary implications

Summary The amino acid sequences of the 139 homologous short neurotoxins, long neurotoxins and cytotoxins so far characterised from elapid snake venoms were compared on the basis of the amino acid deletion/insertion events that have occurred during evolution. Systematic grouping of the toxins according to similarity suggests that the short neurotoxins resemble the cytotoxins more closely than they do the long neurotoxins. The significance of this finding is discussed in relation to the methodology, the conformations of the toxins (as represented by circular dichroism spectra) and the outcome of the study that would have been obtained had more traditional methods been used. It appears probable that the cytotoxins evolved relatively recently from neurotoxic ancestors.     

平颏海蛇毒的毒理学研究

用CM-sephadex C-50柱层析,醋酸铵缓冲液洗脱,从平颏海蛇(Lapemis hardwickii)毒腺提取物中分得15个蛋白组分。其中2个有磷脂酶A_2活性,用免疫学方法证实为肌肉毒组分。其中10个是能作用于突触后膜的神经毒组分。肌肉毒的蛋白质含量约占粗毒蛋白质总量的32.2％。神经毒的蛋白质含量约占粗毒蛋白质总量的52.3％。神经毒的毒性比肌肉毒的毒性强。这些结果提示,平颏海蛇毒有肌肉毒和神经毒两种。神经毒是平颏海蛇毒的主要毒性成分。     

Hainantoxin-Ⅱ的分离纯化及其结构与功能分析(英文)

从分布于我国海南省的海南捕鸟蛛(Haplopelma hainanum)毒素中,利用阳离子交换色谱和反相高效液相色谱分离得到一种多肽神经毒素,命名为Hainantoxin-Ⅱ(HnTx-Ⅱ)。MALDI-TOF质谱分析表明该多肽毒素相对分子质量为4253.135,其氨基酸序列经Edman降解测序为LFECS VSCEI EKEGN KDCKK KKCKG GWKCK FNMCV KV,其中的6个Cys形成3对二硫键。同源性搜索表明,HnTx-Ⅱ与Huwentoxin-Ⅱ(HwTx-Ⅱ)的同源性高达91%,仅有3个氨基酸残基不同。HwTx-Ⅱ为从虎纹捕鸟蛛中提取的杀虫肽,该多肽具有独特的结构模体。活性分析表明HnTx-Ⅱ与HwTx-Ⅱ具有相似的生物学活性。经腹腔注射HnTx-Ⅱ,美洲蜚蠊可被麻痹,其ED50为16μg/g;而当剂量增加到60μg/g时,可立即杀死美洲蜚蠊,其杀死昆虫活性明显强于HwTx-Ⅱ。此外,HnTx-Ⅱ经脑室注射可杀死小鼠,其LD50为1.41μg/g,该活性却明显低于HwTx-Ⅱ。这两种多肽毒素的结构与功能的差异为进一步阐明多肽毒素的结构与功能之间的关系提供良好的研究模型。     

The X-ray structure of a snake venom Gln48 phospholipase A2 at 1.9A resolution reveals anion-binding sites

Phospholipase A2 is an "interfacial" enzyme and its binding to negatively charged surfaces is an important step during catalysis. The Gln48 phospholipase A2 from the venom of Vipera ammodytes meridionalis plays the role of chaperone and directs a toxic His48 PLA2 onto its acceptor. In the venom the two phospholipases A2 exist as a postsynaptic neurotoxic complex, Vipoxin. The X-ray structure of Gln48 PLA2, complexed to sulphate ions, which mimic the negatively charged groups of anionic membranes, has been determined by the molecular replacement method and refined to 1.9A resolution. The protein forms a homodimer stabilized by ionic, hydrophobic, and hydrogen-bond interactions. The structure reveals two anion-binding sites per subunit. These sites are probably involved in interactions with the negatively charged membrane surface and, in this way, in the "targeting" of the toxic component to the receptors of the postsynaptic membranes. In the absence of the chaperone subunit the toxin changes the target of the physiological attack. A comparison of the homodimeric Gln48 PLA2 structure with that of the heterodimeric Vipoxin reveals differences in regions involved in the pharmacological activity of the toxin. This fact, except the active site histidine substitution, can explain the absence of toxicity in the Gln48 protein in comparison to the His48 phospholipase A2.     

Analysis of mobility of protein side chains by spin label technique

Five spin labeled derivatives of a neurotoxin from cobra venom were analyzed by the earlier suggested method. The procedure was adjusted to the complex motional behaviour of the label. Each protein derivative carried covalently bound spin label on different lysine residues. In two derivatives, at positions Lys44 and Lys46, the labels were strongly mobile, whereas for other three derivatives modified at Lys15, Lys25 and Lys26 the label was less mobile with respect to the protein molecule, which made possible determination of the rotational correlation time of the protein molecule (2.8±0.3 ns). The rotational correlation time was in good agreement with the calculated value for the rigid sphere of the corresponding molecular weight. On the basis of the estimate of the anisotropic motion degree, it was found from the order parameter S that the label mobility increases in the following series of lysine residues: Lys26, Lys25, Lys15, Lys46, and Lys44. From the analysis of positions of outer wide peaks in ESR spectra obtained by varying temperature and viscosity of the medium, we determined the parameters for computer simulation. The theoretical and experimental spectra were found to be in good agreement.Nomenclature rotational correlation time of the protein molecule - _l rotational correlation time of the spin label - 2A _Z , 2A the rigid limit distance between OWP and IWP, respectively, for ESR spectra of spin labeled proteins - 2, 2 the averaged limit distance between the OWP and IWP correspondingly mobile spin label to respect of protein moiety with; = - 2A',2A distance between OWP and IWP in the ESR spectra of spin labeled proteins for any T and media Abbreviations SL spin label - NT neurotoxin II from cobra venom - NT-SL-Lys44 neurotoxin spin labeled at Lys44 residue - OWP outer wide peaks in the immobilized ESR spectra - IWP inner wide peaks in the immobilized ESR spectra - WL the residual linewidth     

中华眼镜蛇短链神经毒素cDNA的克隆及在大肠杆菌中的表达

利用RT－PCR技术从中华眼镜蛇毒腺组织中成功地克隆了短链神经毒素CDNA。测序结果表明,该基因开放阅读框架编码83个氨基酸残基,其中对个为信号肽,成熟肽为62个氨基酸残基。该基因与GenBank报道的相同物种的神经毒素基因有相当的同源性,不同物种之间的信号肽序列十分保守。将短链神经毒素CDNA再经PCR扩增除去信号肽序列,克隆到pT7ZZ表达质粒中,转化E.coliBL21（DE3）后,经IPTG诱导可高效表达分子量为23kDa②左右的融合蛋白。表达产物占菌体总蛋白的25％左右。