蝮蛇抗栓酶治疗心房纤颤的体会

我院采用蝮蛇抗栓酶治疗心房纤颤6例,结果阵发性房颤3例痊愈,持续性房颤3例无效.应用蝮蛇抗栓酶治疗阵发性房颤获得良好效果,原因是蝮蛇抗栓酶有降低血浆纤维蛋白原和血液粘度,改善微循环及扩张冠脉等功能,从而改善心肌和心脏传导系统的血液、氧、能量的供应.恢复心脏的正常功能,消除折返激动而终止房颤.     

蝮蛇抗栓酶治疗阳痿2例报告（摘要）

蝮蛇抗栓酶治疗阳痿２例报告（摘要）李殿石,李岐东,尚云凤,段丽霞目前我院在应用蝮蛇抗栓酶（解放军二三八医院产品）治疗脑血管病及高粘滞血症同时,治愈２例阳痿患者,现摘要报告如下：靳××,年龄５７岁,患隙性脑梗塞半年,来院治疗。经血液流变检测结果指标均增...     

蝮蛇抗栓酶与常规法治疗缺血性脑血管病的对比观察

缺血性脑血管病７８例,应用蝮蛇抗栓酶治疗４６例,常规法治疗３２例,有效率分别为９１．３％和８４．３％。认为蝮蛇抗栓酶治疗缺血性脑血管病有较好疗效     

蝮蛇抗栓酶对肺心病急性加重期20例疗效观察（摘要）

蝮蛇抗栓酶对肺心病急性加重期２０例疗效观察（摘要）黑龙江省五常市人民医院林寿源在常规治疗的基础上,应用蝮蛇抗栓酶治疗肺心病急性加重期２０例并与对照组２０例进行疗效对比观察。结果表明,蝮蛇抗栓酶组控制呼吸道感染,心肺功能明显改善,平均需要５—７天,而对...     

蝮蛇抗栓酶治疗血栓性深静脉炎患者7例护理体会

蝮蛇抗栓酶治疗血栓性深静脉炎患者７例护理体会贵州省黔西南州医院神经内科黄晓清１、溶栓治疗的方法及观察本组对７例血栓性深静脉炎患者进行治疗,疗效满意,治愈６例。用蝮蛇抗栓酶０．５ｕ加入０．９％生理盐水１５０ｍｌ中,于患肢踝关节以下静脉滴入,３０～４０滴...     

蝮蛇抗栓酶配合高压氧治疗脑血栓形成的护理体会

我院从 1 989年开始应用蝮蛇抗栓酶配合高压氧治疗脑血栓形成患者 66例 ,效果满意 ,现将护理体会报告如下。1　临床资料　　本组 66例 ,男 36例 ,女 30例 ,年龄最大 76岁 ,最小 42岁 ,平均 58岁。病程最长 6年 ,最短4h。所有病例均用蝮蛇抗栓酶配合高压氧治疗。结果治愈 42例占 63.7% ,好转 2 1例 ,占 31 .8% ,效果不明显 3例 ,占 4.5%。2　护理体会2 .1　治疗操作中的护理问题　蝮蛇抗栓酶临床应用有效剂量 ,成人每天 0 .5～ 0 .75u加入 5%葡萄糖 2 50 ml中静滴 ,每天 1次 ,1 0～ 1 5天为 1疗程 ,一般 2～ 3个疗程后 ,即可收到满意疗效。应…     

蝮蛇抗栓酶治疗不稳定心绞痛疗效分析

蝮蛇抗栓酶已广泛用于治疗缺血性脑血管疾病 ,但用于治疗不稳定心绞痛的临床报道较少。为了观察蝮蛇抗栓酶治疗不稳定心绞痛疗效 ,作者用蝮蛇抗栓酶加常规药物治疗 3 4例不稳定心绞痛患者 ,并与用常规药物治疗的 3 4例不稳定心绞痛患者的疗效进行比较 ,并研究其对临床实验室指标的影响。1　临床资料1 .1　一般资料　按 1 979年 WHO推荐的及 1 994年南京召开的有关不稳定心绞痛诊断治疗研讨会诊断标准 ,并行心电图及心肌酶检查 ,除外急性心肌梗死 ,及使用蝮蛇抗栓酶禁忌症者 ,选出 6 8例不稳定心绞痛患者 ,随机分为两组。蝮蛇抗栓酶组3 4例…     

蝮蛇抗栓酶引起皮下出血2例报告（摘要）

蝮蛇抗栓酶引起皮下出血２例报告（摘要）泉州市人民医院林丽玉,伍淑瑛我院应用蝮蛇抗栓酶治疗脑血栓６４例中,发现有２例治疗过程中出现皮下出血斑,该２例为男性,均为急性脑血栓形成,入院时查血尿便常规及血小板,出凝血时间及纤维蛋白原均在正常范围,蝮蛇抗栓酶每...     

蝮蛇抗栓酶鞘内注入治疗小儿脑病

蝮蛇抗栓酶鞘内注入治疗小儿脑病解放军总医院小儿内科钟炎皋,滕燕蝮蛇抗栓酶已广泛用于治疗心、脑血管等疾病,用于鞘内注入治疗小儿神经系统疾病报道甚少,近２年应用蝮蛇抗栓酶鞘内注入治疗小儿各种脑病１３例,取得较好的疗效,有效率为７６．９０％,现将结果报道如...     

蝮蛇抗栓酶胶囊治疗小卒中263例疗效观察

1994年以来 ,我院应用蝮蛇抗栓酶胶囊 (下称抗栓酶胶囊 )治疗小卒中 (下称 TIA) 2 6 3例 ,疗效显著 ,现总结报告如下。1　临床资料1.1　一般资料　本组 2 6 3例均符合解放军总后勤部卫生部制订的《临床疾病诊断依据 ,治愈好转标准》中 TIA发作的诊断标准。其中男 1 87例 ,女 76例。年龄 3 8～ 77岁 ,其中年龄 3 8～ 49岁 3 5例 ,50～ 6 9岁 1 98例 ,70岁以上 3 0例。病程最短 2 h,最长 2年左右 ,多为反复发作。原有脑动脉硬化 78例 ,有高血压病史 1 2 5例 ,有冠心病 2 5例。1 .2　治疗方法　应用蝮蛇抗栓酶 0 .5～ 0 .75u ( 2～ 3支 )改…     

蝮蛇抗栓酶治疗急性黄疸型肝炎36例临床疗效观察

病毒性肝炎是我国最常见多发的传染病之一,且无特效的药物治疗。本文以前瞻性的随机对照应用蝮蛇抗栓酶0.5单位加入5%葡萄糖250ml 静脉滴注,每日1次,共30天;强力宁80ml加入5%葡萄糖250ml 静脉滴注,每日1次,共30天治疗急性黄疸型肝炎各36例。结果治疗组和对照组总有效率分别为97.2%和94.4%(P>0.05,),以上结果说明,蝮蛇抗栓酶与强力宁治疗急性黄疸型肝炎具有同等疗效.在疗程中未出现出血、过敏和白细胞下降等副反应。     

蝮蛇抗栓酶与精制蝮蛇抗栓酶制剂的比较研究

本文作者对以东北长白山白眉蝮蛇毒为原料生产的两种酶制剂——蝮蛇抗栓酶与精制蝮蛇抗栓酶进行了比较研究。用 HPLC 柱层析粗毒得到15个蛋白峰,同时对两种酶制剂以 HPLC 层析用两根层析柱串联上样层析以提高其分辩率,得到两个图谱,蝮蛇抗栓酶有7个蛋白峰,而精制品有3～4个蛋白峰.同时以 SDS-PAGE 电泳图谱.蝮蛇抗栓酶呈现7～8条谱带,精制品有4～5条谱带,并与已知分子量的标准蛋白进行对比,结果表明两种酶制剂均非单一组分.蝮蛇抗栓酶谱带较多,精制品较纯.其分子量均在2～6万之间.蝮蛇抗栓酶几个组分有协同作用,每次剂量在0.25～0.50单位,即有明显疗效,而精制品临床用药剂量较大,每次0.75～1.0单位,多者达1.25单位(3～5支).     

Detection of Venom after Antivenom Is Not Associated with Persistent Coagulopathy in a Prospective Cohort of Russell's Viper (Daboia russelii) Envenomings

Background

Venom recurrence or persistence in the circulation after antivenom treatment has been documented many times in viper envenoming. However, it has not been associated with clinical recurrence for many snakes, including Russell''s viper (Daboia spp.). We compare the recovery of coagulopathy to the recurrence or persistence of venom in patients with Russell''s viper envenoming.

Methodology/Principal Findings

The study included patients with Russell''s viper (D. russelii) envenoming presenting over a 30 month period who had Russell''s viper venom detected by enzyme immunoassay. Demographics, information on the snake bite, and clinical effects were collected for all patients. All patients had serum collected for venom specific enzyme immunoassay and citrate plasma to measure fibrinogen levels and prothrombin time (international normalised ratio; INR). Patients with venom recurrence/persistence were compared to those with no detectable recurrence of venom. There were 55 patients with confirmed Russell''s viper envenoming and coagulopathy with low fibrinogen concentrations: 31 with venom recurrence/persistence, and 24 with no venom detected post-antivenom. Fibrinogen concentrations increased and INR decreased after antivenom in both the recurrence and non-recurrence patients. Clinical features, laboratory parameters, antivenom dose and length of hospital were similar for both groups. Pre-antivenom venom concentrations were higher in patients with venom recurrence/persistence with a median venom concentration of 385 ng/mL (16–1521 ng/mL) compared to 128 ng/mL (14–1492 ng/mL; p = 0.008).

Conclusion

Recurrence of Russell''s viper venom was not associated with a recurrence of coagulopathy and length of hospital stay. Further work is required to determine if the detection of venom recurrence is due to the venom specific enzyme immunoassay detecting both venom-antivenom complexes as well as free venom.     

Coagulation of plasma from the chicken (Gallus domesticus): phospholipids influence clotting rates induced by components from Russell's viper venom

Added phospholipid failed to accelerate chicken-plasma coagulation, induced by high concentrations of crude Russell's viper venom; however, similarly induced coagulation of canine and human plasma proceeded more rapidly when phospholipid was added. Phospholipid reduced clotting times of canine, human and also chicken plasma when partially purified factor X-activating enzyme from Russell's viper venom was the inducing agent. In the absence of added phospholipid, preincubation of chicken plasma with factor V-activating enzyme from Russell's viper venom accelerated factor X-activating-enzyme-induced coagulation. Preincubation of chicken plasma with the factor V-activating enzyme slowed factor X-activating-enzyme-induced coagulation in the presence of added phospholipid.     

Actin-inhibition and folding of vertebrate deoxyribonuclease I are affected by mutations at residues 67 and 114

Amino acid (aa) residues (Val-67 and Ala-114) have been suggested as being mainly responsible for actin-binding in human and bovine deoxyribonucleases I (DNase I). This study presents evidence of these two aa mutational mechanisms, not only for actin-binding but also for folding of DNase I in mammals, reptiles and amphibians. Human and viper snake (Agkistrodon blomhoffii) enzymes are inhibited by actin, whereas porcine, rat snake (Elaphe quadrivirgata), and African clawed frog (Xenopus laevis) enzymes are not. To investigate the role of aa at 67, mutants of rat snake (Ile67Val) and viper snake (Val67Ile) enzymes were constructed. After substitution, the rat snake was inhibited by actin, while the viper snake was not. For the role of aa at 114, mutants of viper snake (Phe114Ala), rat snake (Phe114Ala), African clawed frog (Phe114Ala), and porcine (Ser114Ala/Ser114Phe) enzymes were constructed. Strikingly, the substitute mutants for viper snake, rat snake and African clawed frog expressed no protein. The porcine (Ser114Ala) enzyme was inhibited by actin, but not the porcine (Ser114Phe) enzyme. These results suggest that Val-67 may be essential for actin-binding, that Phe-114 may be related to the folding of DNase I in reptiles and amphibians, and that Ala-114 may be indispensable for actin-binding in mammals.     

Purification and Functional Characterisation of Rhinocerase,a Novel Serine Protease from the Venom of Bitis gabonica rhinoceros

Background

Serine proteases are a major component of viper venoms and are thought to disrupt several distinct elements of the blood coagulation system of envenomed victims. A detailed understanding of the functions of these enzymes is important both for acquiring a fuller understanding of the pathology of envenoming and because these venom proteins have shown potential in treating blood coagulation disorders.

Methodology/Principal Findings

In this study a novel, highly abundant serine protease, which we have named rhinocerase, has been isolated and characterised from the venom of Bitis gabonica rhinoceros using liquid phase isoelectric focusing and gel filtration. Like many viper venom serine proteases, this enzyme is glycosylated; the estimated molecular mass of the native enzyme is approximately 36kDa, which reduces to 31kDa after deglycosylation. The partial amino acid sequence shows similarity to other viper venom serine proteases, but is clearly distinct from the sequence of the only other sequenced serine protease from Bitis gabonica. Other viper venom serine proteases have been shown to exert distinct biological effects, and our preliminary functional characterization of rhinocerase suggest it to be multifunctional. It is capable of degrading α and β chains of fibrinogen, dissolving plasma clots and of hydrolysing a kallikrein substrate.

Conclusions/Significance

A novel multifunctional viper venom serine protease has been isolated and characterised. The activities of the enzyme are consistent with the known in vivo effects of Bitis gabonica envenoming, including bleeding disorders, clotting disorders and hypotension. This study will form the basis for future research to understand the mechanisms of serine protease action, and examine the potential for rhinocerase to be used clinically to reduce the risk of human haemostatic disorders such as heart attacks and strokes.     

Novel purification strategy for human PON1 and inhibition of the activity by cephalosporin and aminoglikozide derived antibiotics

Human serum paraoxonase (PON1, EC 3.1.8.1.) is a high-density lipid (HDL)-associated, calcium-dependent enzyme; its physiological substrates are not known. In this study, a new purification strategy for human PON1 enzyme was developed using two-step procedures, namely ammonium sulfate precipitation and sepharose-4B-l-tyrosine-1-napthylamine hydrophobic interaction chromatography. SDS-polyacrylamide gel electrophoresis of the enzyme indicates a single band with an apparent MW of 43 kDa. Overall purification rate of our method was found 227-fold. The V(max) and K(m) of the purified enzyme were determined 227.27 EU and 4.16 mM, respectively. The in vitro effects of commonly used antibiotics, namely gentamycin sulfate and cefazolin sodium was also investigated on the purified human serum PON1 enzyme and human liver PON1 enzyme from human hepatoma cell (HepG2). Gentamycin sulfate and cefazolin sodium caused a dose- and time-dependent decrease on PON1 activity in HepG2 cells. Moreover, gentamycin sulfate and cefazolin sodium were effective inhibitors on purified human serum PON1 activity with IC(50) of 0.887 and 0.0084 values, respectively. The kinetics of interaction of gentamycin sulfate and cefazolin sodium with the purified human serum PON1 indicated a different inhibition pattern. Cefazolin sodium showed a competitive inhibition with K(i) of 0.012+/-0.00065 mM. However, Gentamycin sulfate was inhibited in non-competitive manner with K(i) of 0.026+/-0.015. In order to determine the inhibition statue of these drugs on a living system, the effects of same antibiotics on PON1 enzyme activity of mouse serum PON1 and liver PON1 were investigated in vivo. Gentamycin sulfate (3.2 mg/kg) and cefazolin sodium (106.25 mg/kg) leads to the significant decrease in mouse serum PON1 after 2, 4, 6 h and 2, 4 h drug administration, respectively. Cefazolin sodium did not exhibit any inhibition effect for the liver PON1, in vivo.     

DNA Polymerase in Virions of a Reptilian Type C Virus

A study was made of the DNA polymerase of reptilian type C virus isolated from Russell's viper spleen cells. Simultaneous detection experiments demonstrated the presence of 70S RNA and RNA-dependent DNA polymerase activity in reptilian type C virions. The endogenous activity was dependent on the addition of all four deoxynucleotide triphosphates and demonstrated an absolute requirement for a divalent cation. The reptilian viral DNA polymerase elutes from phosphocellulose at 0.22 M salt. In this respect, it is similar to the avian (avian myeloblastosis virus; AMV) viral enzyme but is different from the mammalian (Rauscher leukemia virus; RLV) viral enzyme which elutes at 0.4 M salt. The molecular weight of the viper DNA polymerase as estimated from glycerol gradient centrifugation is 109,000. It is a smaller enzyme than the AMV DNA polymerase (180,000 daltons) and somewhat larger than the RLV enzyme (70,000 daltons). A comparison of other properties of the type C reptilian DNA polymerase with the enzyme found in other type C oncogenic viruses is made.     

Modeling studies on phospholipase A2-inhibitor complexes

Phospholipase A2 (PLA2) is a ubiquitous enzyme that specifically catalyzes hydrolysis of membrane phospholipids to produce lysophospholipids and free fatty acid, namely arachidonic acid, which provides substrate for eicosanoids biosynthesis. Thus, the compounds inhibiting PLA2 have been implicated as potential therapeutic agents in treatment of inflammation related diseases. Plant and marine organisms serve as sources of compounds that act as potential therapeutic agents for treatment of various diseases. The present study reveals the relationship between the structure and function of the medicinally important herbal compounds (acalyphin, chlorogenic acid, stigmasterol, curcumin and tectoridin) and marine compounds (gracilin A and aplysulphurin A). To understand the binding mechanisms of these compounds, molecular modeling studies has been performed with Russell's viper and bovine pancreatic PLA2 as target molecules using molecular operating environment (MOE) software. These compounds show favorable interactions with the amino acid residues at the active site of Russell's viper and bovine pancreatic PLA2, thereby substantiating their proven efficacy as anti-inflammatory compounds and antidotes.     

A possible approach to the analysis of protease mixtures

A problem of quantitative assay of proteases in complex mixtures is solved by using a set of peptide substrates with detectable (chromogenic or fluorogenic) groups (DGs). Quantitation of separate DGs released in reaction of enzyme mixture with a set of substrates is carried out in chromatographic analysis of reaction products. Reaction of peptide derivatives of aminonaphthalene sulfamides with a mixture of thrombin and proteases from viper venom shows the amounts of produced DGs to be proportional to the amounts of both thrombin and venom proteases, confirming the validity of proposed approach. There are cases of mutual influence of some components in proteases mixtures as illustrated by inhibition of trypsin activity in presence of viper venom; one determines enzyme activities in this specific mixture rather than their amounts.