Scolopendra subspinipes mutilans protected the ceruleininduced acute pancreatitis by inhibiting high-mobility group box protein-1

AIM:To evaluate the inhibitory effects of Scolopendra subspinipes mutilans(SSM) on cerulein-induced acute pancreatitis(AP) in a mouse model.METHODS:SSM water extract(0.1,0.5,or 1 g/kg) was administrated intraperitoneally 1 h prior to the first injection of cerulein.Once AP developed,the stable cholecystokinin analogue,cerulein was injected hourly,over a 6 h period.Blood samples were taken 6 h later to determine serum amylase,lipase,and cytokine levels.The pancreas and lungs were rapidly removed for morphological examination,myeloperoxidase assay,and real-time reverse transcription polymerase chain reaction.To specify the role of SSM in pancreatitis,the pancreatic acinar cells were isolated using collagenase method.Then the cells were pre-treated with SSM,then stimulated with cerulein.The cell viability,cytokine productions and high-mobility group box protein-1(HMGB-1) were measured.Furthermore,the regulating mechanisms of SSM action were evaluated.RESULTS:The administration of SSM significantly attenuated the severity of pancreatitis and pancreatitis associated lung injury,as was shown by the reduction in pancreatic edema,neutrophil infiltration,vacuolization and necrosis.SSM treatment also reduced pancreatic weight/body weight ratio,serum amylase,lipase and cytokine levels,and mRNA expression of multiple inflammatory mediators such as tumor necrosis factor-α and interleukin-1β.In addition,treatment with SSM inhibited HMGB-1 expression in the pancreas during AP.In accordance with in vivo data,SSM inhibited the cerulein-induced acinar cell death,cytokine,and HMGB-1 release.SSM also inhibited the activation of c-Jun NH2-terminal kinase,p38 and nuclear factor(NF)-κB.CONCLUSION:These results suggest that SSM plays a protective role during the development of AP and pancreatitis associated lung injury via deactivating c-Jun NH2-terminal kinase,p38 and NF-κB.

Antihepatoma peptide,scolopentide,derived from the centipede scolopendra subspinipes mutilans

BACKGROUND Centipedes have been used to treat tumors for hundreds of years in China.However,current studies focus on antimicrobial and anticoagulation agents rather than tumors.The molecular identities of antihepatoma bioactive components in centipedes have not yet been extensively investigated.It is a challenge to isolate and characterize the effective components of centipedes due to limited peptide purification technologies for animal-derived medicines.AIM To purify,characterize,and synthesize the bioactive components with the strongest antihepatoma activity from centipedes and determine the antihepatoma mechanism.METHODS An antihepatoma peptide(scolopentide)was isolated and identified from the centipede scolopendra subspinipes mutilans using a combination of enzymatic hydrolysis,a Sephadex G-25 column,and two steps of high-performance liquid chromatography(HPLC).Additionally,the CCK8 assay was used to select the extracted fraction with the strongest antihepatoma activity.The molecular weight of the extracted scolopentide was characterized by quadrupole time of flight mass spectrometry(QTOF MS),and the sequence was matched by using the Mascot search engine.Based on the sequence and molecular weight,scolopentide was synthesized using solid-phase peptide synthesis methods.The synthetic scolopentide was confirmed by MS and HPLC.The antineoplastic effect of extracted scolopentide was confirmed by CCK8 assay and morphological changes again in vitro.The antihepatoma effect of synthetic scolopentide was assessed by the CCK8 assay and Hoechst staining in vitro and tumor volume and tumor weight in vivo.In the tumor xenograft experiments,qualified model mice(male 5-week-old BALB/c nude mice)were randomly divided into 2 groups(n=6):The scolopentide group(0.15 mL/d,via intraperitoneal injection of synthetic scolopentide,500 mg/kg/d)and the vehicle group(0.15 mL/d,via intraperitoneal injection of normal saline).The mice were euthanized by cervical dislocation after 14 d of continuous treatment.Mechanistically,flow cytometry was conducted to evaluate the apoptosis rate of HepG2 cells after treatment with extracted scolopentide in vitro.A Hoechst staining assay was also used to observe apoptosis in HepG2 cells after treatment with synthetic scolopentide in vitro.CCK8 assays and morphological changes were used to compare the cytotoxicity of synthetic scolopentide to liver cancer cells and normal liver cells in vitro.Molecular docking was performed to clarify whether scolopentide tightly bound to death receptor 4(DR4)and DR5.qRT-PCR was used to measure the mRNA expression of DR4,DR5,fas-associated death domain protein(FADD),Caspase-8,Caspase-3,cytochrome c(Cyto-C),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),x-chromosome linked inhibitor-of-apoptosis protein and Cellular fas-associated death domain-like interleukin-1βconverting enzyme inhibitory protein in hepatocarcinoma subcutaneous xenograft tumors from mice.Western blot assays were used to measure the protein expression of DR4,DR5,FADD,Caspase-8,Caspase-3,and Cyto-C in the tumor tissues.The reactive oxygen species(ROS)of tumor tissues were tested.RESULTS In the process of purification,characterization and synthesis of scolopentide,the optimal enzymatic hydrolysis conditions(extract ratio:5.86%,IC_(50):0.310 mg/mL)were as follows:Trypsin at 0.1 g(300 U/g,centipede-trypsin ratio of 20:1),enzymolysis temperature of 46°C,and enzymolysis time of 4 h,which was superior to freeze-thawing with liquid nitrogen(IC_(50):3.07 mg/mL).A peptide with the strongest antihepatoma activity(scolopentide)was further purified through a Sephadex G-25 column(obtained A2)and two steps of HPLC(obtained B5 and C3).The molecular weight of the extracted scolopentide was 1018.997 Da,and the peptide sequence was RAQNHYCK,as characterized by QTOF MS and Mascot.Scolopentide was synthesized in vitro with a qualified molecular weight(1018.8 Da)and purity(98.014%),which was characterized by MS and HPLC.Extracted scolopentide still had an antineoplastic effect in vitro,which inhibited the proliferation of Eca-109(IC_(50):76.27μg/mL),HepG2(IC_(50):22.06μg/mL),and A549(IC_(50):35.13μg/mL)cells,especially HepG2 cells.Synthetic scolopentide inhibited the proliferation of HepG2 cells(treated 6,12,and 24 h)in a concentration-dependent manner in vitro,and the inhibitory effects were the strongest at 12 h(IC_(50):208.11μg/mL).Synthetic scolopentide also inhibited the tumor volume(Vehicle vs Scolopentide,P=0.0003)and weight(Vehicle vs Scolopentide,P=0.0022)in the tumor xenograft experiment.Mechanistically,flow cytometry suggested that the apoptosis ratios of HepG2 cells after treatment with extracted scolopentide were 5.01%(0μg/mL),12.13%(10μg/mL),16.52%(20μg/mL),and 23.20%(40μg/mL).Hoechst staining revealed apoptosis in HepG2 cells after treatment with synthetic scolopentide in vitro.The CCK8 assay and morphological changes indicated that synthetic scolopentide was cytotoxic and was significantly stronger in HepG2 cells than in L02 cells.Molecular docking suggested that scolopentide tightly bound to DR4 and DR5,and the binding free energies were-10.4 kcal/mol and-7.1 kcal/mol,respectively.In subcutaneous xenograft tumors from mice,quantitative real-time polymerase chain reaction and western blotting suggested that scolopentide activated DR4 and DR5 and induced apoptosis in SMMC-7721 Liver cancer cells by promoting the expression of FADD,caspase-8 and caspase-3 through a mitochondria-independent pathway.CONCLUSION Scolopentide,an antihepatoma peptide purified from centipedes,may inspire new antihepatoma agents.Scolopentide activates DR4 and DR5 and induces apoptosis in liver cancer cells through a mitochondria-independent pathway.

STAT3 Inhibition by Centipede Scolopendra Extract in Liver Cancer HepG2 Cells and Orthotopic Mouse Models of Hepatocellular Carcinoma

Objective To observe the effects of Centipede Scolopendra extraction(CSE)on human liver cancer HepG2 cells and the nude mouse tumor model of liver orthotopic transplantation,and to explore the anti-liver cancer mechanism of the extract.Methods HepG2 cells were respectively treated with CSE250(250μg/mL),CSE500(500μg/mL)and 5-FU,and control group was established.An enzymatic hydrolysis and acetone precipitation method was used to separate and purify CSE,which was then used to treat HepG2 cells.The CCK8 assay was used to detect the inhibition of cell proliferation and the half maximal inhibitory concentration(IC50)was calculated.Flow cytometry was used to analyze the cell cycle,and western blot was used to detect the expression of signal transduction and activator of transcription 3(STAT3)pathway-related proteins in HepG2 cells treated with CSE.A nude mouse model with an orthotopic liver tumor was prepared.The mice were randomly divided into four groups,each containing 12 animals:the model group,the 5-FU group,the CSE10 group[10 mg/(kg·d)]and the CSE50 group[50 mg/(kg·d)].The volume and mass changes in the nude mice with orthotopic transplanted tumors were observed.Western blot method was used to test the protein expression levels of p-STAT3 and p38 mitogen-activated protein kinase(p38MAPK).Tissues from the liver of mice in the model group and the CSE50 group were analyzed by using a protein tyrosine kinase(PTK)chip,and the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)function enrichment analysis of the differentially expressed proteins was performed.Results This study showed that CSE significantly inhibited the proliferation of HepG2 cells(P<0.05).After 48 h of CSE treatment,the cell cycle of HepG2 cells manifested as S phase and G2/M phase;p-STAT3 protein levels in the CSE groups were significantly lower than that in the control group(P<0.05).Analysis of the tumor inhibition in the mice showed that the tumor masses and volume in CSE groups were lower(P<0.05).The protein levels of p-STAT3 and p38MAPK in CSE50 group and 5-FU group decreased significantly(P<0.05).PTK antibody chip screening results showed that CSE groups had a bidirectional regulation trend,and there were 23 up-regulated PTKs and six down-regulated PTKs.The GO and KEGG analyses showed that CSE exerted its anticancer effects through regulation of biological processes,including mitogen-activated protein kinase(MAPK)cascade,chemotaxis,cell invasion,cell adhesion,angiogenesis and other biological processes,and through signaling pathways,including the MAPK,phosphatidylinositol-3-kinase/serine threonine protein kinase(PI3K/AKT),and RAS signaling pathways.Conclusions CSE can effectively inhibite the proliferation of HepG2 cells and effectively inhibite the growth of liver cancer orthotopic transplantation tumor.Its mechanism may be closely related to the regulation of STAT3,MAPK and PI3K/AKT signaling pathways.

Scorpiones,Scolopendra and Gekko Inhibit Lung Cancer Growth and Metastasis by Ameliorating Hypoxic Tumor Microenvironment via PI3K/AKT/mTOR/HIF-1αSignaling Pathway

Objective:To investigate whether Buthus martensii karsch(Scorpiones),Scolopendra subspinipes mutilans L.Koch(Scolopendra)and Gekko gecko Linnaeus(Gekko)could ameliorate the hypoxic tumor microenvironment and inhibit lung cancer growth and metastasis by regulating phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin/hypoxia-inducible factor-1α(PI3K/AKT/mTOR/HIF-1α)signaling pathway.Methods:Male C57BL/6J mice were inoculated with luciferase labeled LL/2-luc-M38 cell suspension to develop lung cancer models,with rapamycin and cyclophosphamide as positive controls.Carboxy methyl cellulose solutions of Scorpiones,Scolopendra and Gekko were administered intragastrically as 0.33,0.33,and 0.83 g/kg,respectively once daily for 21 days.Fluorescent expression were detected every 7 days after inoculation,and tumor growth curves were plotted.Immunohistochemistry was performed to determine CD31 and HIF-1αexpressions in tumor tissue and microvessel density(MVD)was analyzed.Western blot was performed to detect the expression of PI3K/AKT/mTOR/HIF-1αsignaling pathway-related proteins.Enzyme-linked immunosorbent assay was performed to detect serum basic fibroblast growth factor(bFGF),transforming growth factor-β1(TGF-β1)and vascular endothelial growth factor(VEGF)in mice.Results:Scorpiones,Scolopendra and Gekko prolonged the survival time and inhibited lung cancer metastasis and expression of HIF-1α(all P<0.01).Moreover,Scorpiones,Scolopendra and Gekko inhibited the phosphorylation of AKT and ribosomal protein S6 kinase(p70S6K)(P<0.05 or P<0.01).In addition,they also decreased the expression of CD31,MVD,bFGF,TGF-β1 and VEGF compared with the model group(P<0.05 or P<0.01).Conclusion:Scorpiones,Scolopendra and Gekko all showed beneficial effects on lung cancer by ameliorating the hypoxic tumor microenvironment via PI3K/AKT/mTOR/HIF-1αsignaling pathway.

Quinoline alkaloids isolated from Scolopendra subspinipes mutilans

Objective: To study the quinoline alkaloids from the ethanol extract of Scolopendra subspinipes mutilans(SSM).Methods: The chemical constituents were isolated and purified by macroporous resin column, medium pressure preparation chromatography, and semi-preparative HPLC. Their structures were elucidated by IR,MS, and NMR experiments.Results: Three quinolone alkaloids were obtained and identified as 3-hydroxy-4-methoxyquinolin-8-yl hydrogen sulfate(1), jineol-8-sulfate(2), and jineol(3), respectively.Conclusion: Compound 1 is a new compound from SSM.

中国少棘蜈蚣(Scolopendra subspinipes mutillans)的毒腺结构

用免疫组化和光镜、透射电镜等观察了中国少棘蜈蚣毒腺的结构。结果显示,纵贯颚肢的弯月形毒腺为单管泡状腺,主要由柱状分泌细胞和介于其间的纤细表皮细胞组成。被肌肉束环绕的分泌细胞辐射状排列于几丁质的毒液导管外,其纤细的颈部由环状括约肌控制,分泌端以折叠回转的单向瓣膜经导管壁上的孔道直接伸入管腔,膨大的盲端直达毒腺底膜。高电子密度的分泌溶酶体向分泌口汇集时电子密度逐渐降低并降解为分泌小泡,其中的杆状结晶样毒蛋白也经无定型状态逐渐分散,经胞吐作用进入管腔并进一步疏散和均质化。免疫组化显示,分泌细胞颈部密集的分泌颗粒的主要成分为毒蛋白,毒蛋白在分泌细胞中呈明显的向心式梯度增强型分布。根据上述观察,提出了蜈蚣毒液以分泌溶酶体介导的非经典途径分泌的观点。

Synthesis of Novel Scolopendra-type Polydodecyloxybenzoyl[1,5]-diazocine as New Material for Optical Sensor

A new scolopendra-type polymer of polydodecyloxybenzoyl[1,5]-diazocine（PDBD） was designed and prepared using 2,5-bis（4-（dodecyloxy）-benzoyl）terephthaloyl azide with trifluoroacetic acid（TFA） via one-pot reaction in good yields. The structure of polymer was characterized using ~1 H-NMR, ^（13） C-NMR and MALDI-TOF spectra. The polymer PDBD exhibits good thermal stability as measured by TGA and DSC, and can be dissolved well in common organic solvents such as chloroform and tetrahydrofuran. In addition, UV-Vis spectral studies indicate that the polymer PDBD shows unique optical property changes（protonation/deprotonation） in the different trifluoroacetic acid environments. The new polymer is expected to be utilized as an optical functional material for fabricating optical sensors in environmental and biological fields.

蜈蚣药材、饮片、标准汤剂与配方颗粒的特征图谱相关性研究及量值传递分析

目的建立蜈蚣药材、饮片、标准汤剂与配方颗粒的高效液相色谱(HPLC)特征图谱,并测定其中次黄嘌呤、苯丙氨酸成分的含量,考察四者之间化学成分的传递与差异。方法采用Dikma Platisil ODS-C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇和水为流动相梯度洗脱,流速为1 mL·min^(-1),柱温为25℃。以含量、特征图谱共有峰传递数及相似度为主要评价指标,分析其量值传递规律。结果18批蜈蚣药材、饮片、标准汤剂及3批配方颗粒特征图谱均有8个共有特征峰,与各自对照特征图谱相似度均大于0.90,指认出尿嘧啶、酪氨酸、次黄嘌呤、黄嘌呤、苯丙氨酸、肌苷、鸟苷、色氨酸8种成分,并测定其含量,未出现离散数据。结论蜈蚣药材、饮片、标准汤剂与配方颗粒的主要化学成分基本相同,HPLC特征图谱相关性良好,8种成分在测定范围内与峰面积线性关系良好。所建立质量评价模式能够反映蜈蚣4种形态多成分的整体面貌,为蜈蚣配方颗粒质量控制提供数据支撑。

黑头蜈蚣的化学成分

本文报道了黑头蜈蚣Scolopendra negrocapitis化学成分,并与中国药典收载的少棘巨蜈蚣S.subspinipes mutilans进行了比较。黑头蜈蚣含总脂18.7％,蛋白质63.4％,游离氨基酸11.9％;含有与少棘巨蜈蚣相同的12种脂肪酸(其中不饱和脂肪酸占64％),21种氨基酸和12种微量元素;蛋白质聚丙酰胺凝胶电泳分离出14条色带。表明黑头蜈蚣与少棘巨蜈蚣主要化学成分类同,是一种值得开发利用的新药源。

全蝎蜈蚣对CIA大鼠外周血CD4^+CD25^+FoxP3^+Treg细胞的调节

目的:观察全蝎蜈蚣对胶原免疫性关节炎(Collagen-Induced Arthritis,CIA)大鼠外周血CD4+CD25+FoxP3+Treg细胞的调节作用。方法:Wistar大鼠60只,随机分成6组,分别为正常对照组、模型对照组、全蝎蜈蚣高(0.4 g/kg)、中(0.2 g/kg)、低剂量(0.1 g/kg)组和Ⅱ型胶原蛋白(60μg/kg)治疗组(CⅡ组),二型胶原蛋白诱导大鼠关节炎,并采用流式细胞术检测大鼠外周血CD4+CD25+FoxP3+Treg细胞及酶联免疫吸附试验检测血清中IL-2表达水平。结果:与模型组比较,全蝎蜈蚣高、低剂量组大鼠外周血CD4+CD25+T细胞及CD4+CD25+FoxP3+Treg细胞水平显著升高,且中、低剂量组血清IL-2水平显著下降。结论:全蝎蜈蚣治疗关节炎大鼠可能是通过上调外周CD4+CD25+FoxP3+Treg细胞表达水平以恢复或重建免疫耐受而实现的。

少棘蜈蚣纤溶活性蛋白的抗血栓作用

目的研究少棘蜈蚣(Scolopendra subspinipes mutilans L.Koch)纤溶活性蛋白的抗血栓作用。方法采用离子交换层析和分子筛等制备法从少棘蜈蚣中纯化到蜈蚣纤溶酶(Scolopendra subspinipes mutilans L.Koch fibrinolyti cenzyme,SSFE),用纤维蛋白平板检测酶活力,常规方法检测了SSFE溶血性和出血性,进行了SSFE体外溶栓和体内溶栓实验,并检测了凝血酶原时间(PT)、活化部分凝血酶时间(APTT)和凝血酶时间(TT)3个标准化指标。结果PAGE显示SSFE为单一组分,具有纤溶活性;证明SSFE无致出血性且无致溶血活性,低、中、高剂量SSFE使小鼠APTT和TT均明显延长,中剂量SSFE对PT有延长作用,而高剂量作用不明显。结论蜈蚣纤溶酶具有抗血栓作用。

多棘蜈蚣化学成分的研究

目的：分析多棘蜈蚣（ＳｃｏｌｏｐｅｎｄｒａｍｕｌｔｉｄｅｎｓＮｅｗｐｏｒｔ）的化学成分，并与药典收载品少棘蜈蚣（Ｓ．ｍｕｔｉｌａｎｓＬ．Ｋｏｃｈ）比较。方法：采用溶媒提取法测定总脂，气相色谱质谱计算机检测脂肪酸，微量凯氏定氮法测定蛋白质，离子交换色谱测定游离氨基酸，电感耦合离子发射光谱测定微量元素。结果：多棘蜈蚣含总脂７．２％，蛋白质６４．６％，游离氨基酸６．３％；含有与少棘蜈蚣相同的１５种脂肪酸（其中不饱和脂肪酸占７２％）、２１种氨基酸和１２种微量元素。结论：多棘蜈蚣与少棘蜈蚣主要化学成分类同，某些成分含量存在差异，是一种值得开发利用的药用蜈蚣资源。

少棘蜈蚣水提取物的抗菌活性

用含有大肠杆菌Escherichia coli K12D31／poly IC和昆虫生理盐水注射液诱导3～4d的少棘蜈蚣Scolo—pendra subspinipes mutilans水提取物表现出较高的抗菌活性，经测定发现在设定的不同温度、pH值及离子强度条件下表现出良好的抗菌活性；对不同的蛋白酶表现出不同的敏感程度；除了对Proteus mirobilis未表现出抑菌外，对其它9种革兰氏阳性菌和阴性菌、真菌都有抗菌作用，表明有广谱抗菌活性；在600μg/ml以下，没有表现出溶血和凝集活性；电镜观察表明可损伤大肠杆菌的外壁，内容物外泄，菌体死亡。

蜈蚣药材薄层鉴别及抗凝活性定量的研究

目的:对蜈蚣药材的薄层鉴别及抗凝活性定量方法进行研究,为该药材质量控制标准的完善提供依据。方法:对蜈蚣药材中游离精、丝氨酸进行薄层层析,筛选供试品制备方法及展开剂系统;以凝血酶滴定法测定蜈蚣药材抗凝活性,筛选药材前处理方法。结果:蜈蚣药材经甲酸及95%乙醇(1∶1)超声处理后,以正丁醇-乙酸-水(12∶5∶4)体系于硅胶G板上进行薄层层析,经茚三酮显色后,斑点清晰,杂质及拖尾影响较小,目标成分分离度较好;以凝血酶滴定法测定蜈蚣药材抗凝活性,结果重现性较好,超声法提取药材中的抗凝成分,操作简单,耗时较短,所制供试品抗凝血酶活性为(14.00±1.53)U/g;另测3批不同批次蜈蚣药材抗凝活性分别为:(13.00±0.58)U/g、(17.00±1.15)U/g、(15.67±1.53)U/g。结论:所选择的薄层鉴别及抗凝活性定量方法可作为完善蜈蚣药材质量标准的依据。

蜈蚣有效成分抗心肌缺血作用的研究

目的 :观察蜈蚣有效成分对心肌缺血鼠自由基的变化 ,阐明蜈蚣治疗冠心病的机制。方法 :将4 0只小鼠随机分为 4组 ,A组为空白对照组 ,B组为模型组 ,C组为蜈蚣小剂量 ( 2 5g/kg)治疗组 ,D组为蜈蚣大剂量 ( 5g/kg)治疗组。造模前治疗组用不同剂量的蜈蚣提取物 ,分别按每天 0 2mL/ 1 0g灌胃 ,连续 7天 ,其余两组用等量蒸馏水灌胃。后采用脑垂体后叶素 2 0u/kg腹腔注射 ,诱发心肌缺血。缺血 2 0min后 ,观察各组心肌酶、自由基的变化 ,以及心肌透射电镜。结果 :模型组LDH、MDA明显升高 ,SOD、NO水平显著降低 ,治疗组SOD、NO含量显著提高 ,LDH、MDA明显降低 ,超微结构显示心肌细胞损害明显减轻。且蜈蚣大剂量较小剂量治疗组作用更加明显。结论 :蜈蚣具有抗心肌缺血的作用。

纳升级反相液相色谱-串联质谱法分析蜈蚣提取蛋白质

结合聚丙烯酰胺凝胶电泳和纳升级反相液相色谱-串联质谱(nanoRPLC-MS/MS)法系统分析了蜈蚣提取蛋白质胶内酶解和直接酶解产物,并进行数据库检索鉴定蛋白质。建立的nanoLC分析条件为:上样8μL,经Chrom XP-C18毛细管柱(350μm×0.5 mm,3μm),2%乙腈(0.1%甲酸)-98%乙腈(0.1%甲酸)梯度洗脱90 min;采用ESI-Q-TOF MS,并在IDA采集模式下分析鉴定蜈蚣提取蛋白质。胶内酶解鉴定到72种蛋白质,直接酶解鉴定到97种蛋白质,二者含26种相同的蛋白质。通过数据库比对,分析了蜈蚣提取蛋白质的生物进程、细胞组分以及功能活性。得到蜈蚣蛋白质具有结合、酶催化、肌动等功能活性,其中结合活性的蛋白质占的比例较大,且多为能量代谢进程中的相关酶类。

刘伟胜教授运用全蝎、蜈蚣治疗恶性肿瘤经验

介绍名中医刘伟胜教授运用全蝎、蜈蚣治疗恶性肿瘤的经验。指出全蝎、蜈蚣为解毒散结、熄风止痉药,临证治癌宜灵活辨证,配合温阳祛寒,或理气活血,或补气益气,或祛痰散结;注重辨病,结合全蝎、蜈蚣药理,以增强抗癌抑癌之力;审时度势,使用时应把握其用量、用法及用药时间,适可而止。

少棘蜈蚣毒腺RACE cDNA文库的构建及β-actin基因的克隆与序列分析

运用SMART技术首次构建了少棘蜈蚣(Scolopendrasubspinipes)毒腺cDNA文库。经琼脂糖电泳检测,文库所含全长cDNA主要分布在5 0 0bp以上,大于2 0 0 0bp的区域尚有很长拖尾,中间有几条高丰度mRNA亮带。以此文库为模板,通过5′RACE(RapidamplificationofcDNAends,快速扩增cDNA末端)方法获得了细胞质肌动蛋白βactin基因5′末端5 98bp片段,其中包括开放阅读框5 46bp ,编码1 82个氨基酸。将该基因片段推导的氨基酸序列通过BLAST软件与蛋白质公共数据库Swissprot比对,发现与蜜蜂(Apismellifera)的βactin基因同源性高达96% ,说明本实验所构文库质量完全可以满足用RACE方法进行功能基因的cDNA克隆。通过基于双参数模型的NJ法对部分动物βactin基因进行系统重建,较好地反映了这些动物的系统发生关系。

少棘蜈蚣毒液溶血肽的分离纯化

采用超滤和HPLC(包括凝胶过滤层析、阳离子交换层析和反相HPLC)等技术对少棘蜈蚣毒液进行分离纯化,最终获得一个多肽;运用基质辅助激光解吸附飞行时间质谱测定其分子量为4484Da,测得其对小鼠红细胞半数溶血剂量HD50=14.69μg/ml。

墨江蜈蚣与少棘蜈蚣的比较研究──Ⅰ．化学组成

本文对墨江蜈蚣（ＳｃｏｌｏｐｅｎｄｒａｍｏｊａｎｇｉｃａＺｈａｎｇｅｔＣｈｉ）与少棘蜈蚣（ＳｃｏｔｏｐｅｎｄｒａｓｕｂｓｐｉｎｉｐｅｓｍｕｔｉｌａｎｓＬ．Ｋｏｃｈ）的水分、灼烧残渣、蛋白质、游离氨基酸、微量元素和挥发性脂肪酸的含量进行了系统的测定并比较了两者间的异同性。实验结果表明：墨江蜈蚣的蛋白质、灼烧残渣、水分的含量分别为６７．２％、３．９５％和３．５９％，而少棘蜈蚣分别为６８．８％、４．８０％和３．６５％，它们的含量基本相同。游离氨基酸、挥发性脂肪酸和微量元素的相对含量也很接近，但脂类和总糖的含量差异较大。