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Antihepatoma peptide,scolopentide,derived from the centipede scolopendra subspinipes mutilans 被引量:4
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作者 Yu-Xing Hu Zhuo Liu +11 位作者 Zhen Zhang Zhe Deng Zhen Huang Ting Feng Qing-Hong Zhou Si Mei Chun Yi Qing Zhou Pu-Hua Zeng Gang Pei Sha Tian Xue-Fei Tian 《World Journal of Gastroenterology》 SCIE CAS 2023年第12期1875-1898,共24页
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... 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. 展开更多
关键词 scolopendra centipede Antihepatom peptide Hepatocellular carcinoma Death receptor 4 Death receptor 5
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Scolopendra subspinipes mutilans protected the ceruleininduced acute pancreatitis by inhibiting high-mobility group box protein-1 被引量:7
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作者 Il-Joo Jo Gi-Sang Bae +7 位作者 Kyoung-Chel Park Sun Bok Choi Won-Seok Jung Su-Young Jung Jung-Hee Cho Mee-Ok Choi Ho-Joon Song Sung-Joo Park 《World Journal of Gastroenterology》 SCIE CAS 2013年第10期1551-1562,共12页
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 intraperitonea... 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. 展开更多
关键词 scolopendra subspinipes mutilans CYTOKINES Acute PANCREATITIS HIGH-MOBILITY GROUP box protein-1
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STAT3 Inhibition by Centipede Scolopendra Extract in Liver Cancer HepG2 Cells and Orthotopic Mouse Models of Hepatocellular Carcinoma 被引量:7
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作者 TENG Yong-Jie LIU Zhuo +3 位作者 LIAO Liu CHEN Yuan HUANG Xiao-Di TIAN Xue-Fei 《Digital Chinese Medicine》 2020年第2期67-79,共13页
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 mechan... 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. 展开更多
关键词 centipede scolopendra extract(CSE) Liver cancer Nude mice Protein tyrosine kinase(PTK) STAT3 Protein chip
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Clinical consequences of centipede bite:Is it neurotoxic?
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作者 Ioannis N Mavridis Maria Meliou Efstratios-Stylianos Pyrgelis 《World Journal of Neurology》 2016年第2期23-29,共7页
The primary purpose of this article was to review the current literature regarding the clinical consequences of centipede envenomation in humans,in order to determine whether the bite of these arthropods is neurotoxic... The primary purpose of this article was to review the current literature regarding the clinical consequences of centipede envenomation in humans,in order to determine whether the bite of these arthropods is neurotoxic to humans or not. A thorough search of the literature regarding the clinical consequences of centipede bites in humans was applied,with great respect to neurological symptoms potentially caused by such bites. Centipede bite commonly causes only local reactions,which usually resolve within a few days without sequelae. The patients in the majority of centipede envenomations describe a painful but benign syndrome. However,mild constitutional symptoms are relatively frequent. Remarkably,centipedes can rarely cause severe systematic reactions such as anaphylaxis or even hypotension and myocardial ischemia. Factors such as patient age,comorbidity,anatomic site of envenomation,and size/species of centipede should be considered when evaluating a centipede envenomation victim. According to the current literature,the centipede bite does not seem to be neurotoxic to humans. However,it commonly causes symptoms mediated by the nervous system. These include local and generalized symptoms,with the first dominated by sensory disturbances and the second by non-specific symptoms such as headache,anxiety and presyncope. Based on our results,the answer to our study's question is negative. The centipede bite is not neurotoxic to humans. However,it commonly causes symptoms mediated by the nervous system,which include primarily local pain and sensory disturbances,as well as generalized non-specific symptoms such as headache,anxiety and vagotonia. 展开更多
关键词 centipedeS Pain Sensory disturbances ENVENOMATION scolopendra
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中国少棘蜈蚣(Scolopendra subspinipes mutillans)的毒腺结构
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作者 华卫建 徐国华 +1 位作者 周祀乔 华子春 《中国细胞生物学学报》 CAS CSCD 北大核心 2012年第2期162-167,共6页
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... 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。 展开更多
关键词 少棘蜈蚣 毒腺 显微结构 超微结构 免疫组化 分泌溶酶体
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Quinoline alkaloids isolated from Scolopendra subspinipes mutilans 被引量:2
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作者 Yan-fang Li Wu-zhan Liu +4 位作者 Jian-wei Fan Chuan-liang Huang Li-hua Deng Hui-fang Zhuang Yong-xia Guan 《Chinese Herbal Medicines》 CAS 2019年第3期344-346,共3页
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 ... 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. 展开更多
关键词 3–hydroxy-4-methoxyquinolin-8-yl hydrogen SULFATE jineol jineol-8-sulfate QUINOLINE ALKALOIDS scolopendra subspinipes mutilans L. Koch
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黑头蜈蚣的化学成分 被引量:22
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作者 方红 邓芬 +1 位作者 严宜昌 王克勤 《中药材》 CAS CSCD 1999年第5期226-228,共3页
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... 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。 展开更多
关键词 黑头蜈蚣 少棘巨蜈蚣 化学成分 蜈蚣
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少棘蜈蚣纤溶活性蛋白的抗血栓作用 被引量:53
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作者 陈少鹏 韩雅莉 +1 位作者 郭桅 李兴暖 《中国药理学通报》 CAS CSCD 北大核心 2007年第8期1088-1092,共5页
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... 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。 展开更多
关键词 蜈蚣 纤溶活性蛋白 抗血栓作用
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少棘蜈蚣水提取物的抗菌活性 被引量:19
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作者 任文华 张双全 +1 位作者 宋大祥 周开亚 《中药材》 CAS CSCD 北大核心 2007年第1期10-14,共5页
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... 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。 展开更多
关键词 少棘蜈蚣 抗菌活性 抑菌谱 溶血 凝集
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蜈蚣药材薄层鉴别及抗凝活性定量的研究 被引量:19
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作者 李桃 谭晓梅 +1 位作者 龙群 陈飞龙 《中药材》 CAS CSCD 北大核心 2012年第5期686-689,共4页
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... 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。 展开更多
关键词 蜈蚣 薄层鉴别 凝血酶滴定 质量
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少棘蜈蚣毒液溶血肽的分离纯化 被引量:5
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作者 任文华 张双全 +1 位作者 宋大祥 周开亚 《动物学报》 SCIE CAS CSCD 北大核心 2007年第3期519-523,共5页
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。
关键词 少棘蜈蚣 毒液 溶血肽 分离纯化
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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 被引量:4
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作者 任文华 张双全 +1 位作者 宋大祥 周开亚 《动物学杂志》 CAS CSCD 北大核心 2005年第3期1-5,共5页
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(Rapidamp... 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。 展开更多
关键词 cDNA文库 RACE 少棘蜈蚣 n基因 序列分析 毒腺 构建 快速扩增CDNA末端 SMART技术 BLAST软件 全长cDNA cDNA克隆 系统发生关系 开放阅读框 公共数据库 氨基酸序列 基因同源性 双参数模型 电泳检测 mRNA 肌动蛋白 基因片段
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墨江蜈蚣与少棘蜈蚣的比较研究──Ⅰ.化学组成 被引量:4
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作者 冉永禄 吴刚 +2 位作者 叶文娟 迟程 罗天诰 《Zoological Research》 CAS CSCD 1995年第4期379-383,共5页
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... 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。 展开更多
关键词 墨江蜈蚣 少棘蜈蚣 化学组成
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中药蜈蚣多糖的分离纯化及性质研究 被引量:8
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作者 李兴暖 肖湘 王云 《中药材》 CAS CSCD 北大核心 2009年第6期846-848,共3页
目的:从中药蜈蚣体内提取多糖组分,并对其部分性质进行分析。方法:从中药蜈蚣体内提取粗多糖,经DEAE-52离子交换柱层析分离获得3个糖组分;凝胶过滤法测组分Ⅰ的分子量;MTT法检测多糖组分Ⅰ对两种肿瘤细胞的抑制作用。结果:组分Ⅰ的分子... 目的:从中药蜈蚣体内提取多糖组分,并对其部分性质进行分析。方法:从中药蜈蚣体内提取粗多糖,经DEAE-52离子交换柱层析分离获得3个糖组分;凝胶过滤法测组分Ⅰ的分子量;MTT法检测多糖组分Ⅰ对两种肿瘤细胞的抑制作用。结果:组分Ⅰ的分子量为33.1kD;该多糖组分对Hela细胞的抑制作用明显,当糖浓度为3.13μg/mL时,抑制率达60.8%,但对Eca109细胞无明显抑制作用。结论:蜈蚣多糖组分Ⅰ对不同的肿瘤细胞具有特异性。 展开更多
关键词 蜈蚣 多糖 分离纯化 肿瘤细胞
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蜈蚣多糖对Hela细胞的增殖抑制作用研究 被引量:16
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作者 李兴暖 韩雅莉 余卫国 《时珍国医国药》 CAS CSCD 北大核心 2009年第7期1571-1573,共3页
目的研究蜈蚣多糖对人宫颈癌细胞Hela细胞的抑制作用及作用机制。方法用MTT法检测蜈蚣多糖对Hela细胞生长的抑制作用;流式细胞仪和AnnexinV-FITC/PI双荧光染色分别检测蜈蚣多糖对Hela细胞周期及凋亡情况的影响。结果在一定浓度范围内蜈... 目的研究蜈蚣多糖对人宫颈癌细胞Hela细胞的抑制作用及作用机制。方法用MTT法检测蜈蚣多糖对Hela细胞生长的抑制作用;流式细胞仪和AnnexinV-FITC/PI双荧光染色分别检测蜈蚣多糖对Hela细胞周期及凋亡情况的影响。结果在一定浓度范围内蜈蚣多糖可显著抑制Hela细胞的增殖,改变Hela细胞的细胞周期,使细胞阻滞于G2/M期,并能诱导细胞凋亡。结论蜈蚣多糖可能通过改变Hela细胞的细胞周期,诱导细胞凋亡,从而对Hela细胞的增殖有一定的抑制作用。 展开更多
关键词 蜈蚣多糖 细胞周期 细胞凋亡
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蜈蚣醇提取物和水提取物部分药理作用比较 被引量:6
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作者 周莉莉 黄迎春 任超 《时珍国医国药》 CAS CSCD 北大核心 2008年第11期2697-2698,共2页
目的比较鲜品少棘蜈蚣的醇提物及水提物在体外抑菌和抗惊厥两方面的作用。方法采用平板空穴扩散法,以大肠杆菌、金黄色葡萄球菌、枯草杆菌黑色变种芽孢、白色念珠球菌、黑曲霉菌5种常见菌为供试菌,对蜈蚣醇提物及水提物的抑菌活性进行研... 目的比较鲜品少棘蜈蚣的醇提物及水提物在体外抑菌和抗惊厥两方面的作用。方法采用平板空穴扩散法,以大肠杆菌、金黄色葡萄球菌、枯草杆菌黑色变种芽孢、白色念珠球菌、黑曲霉菌5种常见菌为供试菌,对蜈蚣醇提物及水提物的抑菌活性进行研究;采用硝酸士的宁致痫模型,观察蜈蚣醇提物及水提物对小鼠抗惊率、死亡率及惊厥潜伏期的影响。结果蜈蚣醇提物水提物对各供试菌均有抑制作用,醇提物抑菌效果明显强于水提物。蜈蚣水提物能够明显提高抗惊率,降低死亡率,延长小鼠惊厥潜伏期,潜伏期达到(372±146.9)s,与对照组比较有显著性差异(P<0.01)。蜈蚣醇提物没有明显抗惊厥作用。结论蜈蚣活性成分及作用效果与提取部位密切相关。 展开更多
关键词 少棘蜈蚣 提取物 抑菌活性 抗惊厥作用
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蜈蚣不同部位抗凝活性的研究 被引量:28
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作者 刘瑞连 廖建萍 刘绍贵 《中医药导报》 2015年第2期32-33,共2页
目的:比较蜈蚣不同部位提取方法抗凝活性的强弱。方法:用凝血酶时间为指标测定冷浸法和水提醇沉法对蜈蚣不同部位的抗凝血活性。结果:水提醇沉法蜈蚣凝血酶时间长于冷浸法,两种提取方法中,各部位的凝血时间比较,顺序是蜈蚣身>蜈蚣全... 目的:比较蜈蚣不同部位提取方法抗凝活性的强弱。方法:用凝血酶时间为指标测定冷浸法和水提醇沉法对蜈蚣不同部位的抗凝血活性。结果:水提醇沉法蜈蚣凝血酶时间长于冷浸法,两种提取方法中,各部位的凝血时间比较,顺序是蜈蚣身>蜈蚣全体>蜈蚣尾。结论:蜈蚣抗凝活性成分提取方法中水提醇沉优于冷浸法,抗凝活性成分主要集中于蜈蚣身。 展开更多
关键词 蜈蚣 提取方法 凝血酶时间
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蜈蚣及其混淆品的鉴别研究 被引量:6
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作者 杨成俊 周伟庆 《安徽农业科学》 CAS 北大核心 2011年第25期15402-15403,共2页
[目的]对蜈蚣及其混淆品的性状与凝胶电泳图谱进行研究。[方法]采用SDS-聚丙烯酰胺凝胶电泳法,测定并比较它们的电泳图的谱带数目和种类。[结果]蜈蚣及其混淆品的电泳谱带数目和种类均不相同。[结论]SDS-PAGE图谱可以作为蜈蚣及其混淆... [目的]对蜈蚣及其混淆品的性状与凝胶电泳图谱进行研究。[方法]采用SDS-聚丙烯酰胺凝胶电泳法,测定并比较它们的电泳图的谱带数目和种类。[结果]蜈蚣及其混淆品的电泳谱带数目和种类均不相同。[结论]SDS-PAGE图谱可以作为蜈蚣及其混淆品的有效鉴别依据。 展开更多
关键词 少棘巨蜈蚣 多棘蜈蚣 墨江蜈蚣 燕山蛩 十二烷基硫酸钠聚丙烯酰胺凝胶电泳
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墨江蜈蚣与少棘蜈蚣对致病真菌和细菌体外生长的影响 被引量:5
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作者 吴刚 王金焕 +3 位作者 冉永禄 迟程 叶文娟 唐向辉 《皮肤病与性病》 北大核心 1992年第2期12-14,共3页
本文分析了少棘和墨江蜈蚣对8种致病真菌和7种细菌体外生长的影响。两种蜈蚣的2%NaHCO_3和蒸馏水提取液以及油脂对真菌影响极小;3%CH_3COOH 提取液强烈抑制真菌生长,但无杀菌作用,测定了相应的最小抑菌浓度(MIC)。各提取物对细菌均无... 本文分析了少棘和墨江蜈蚣对8种致病真菌和7种细菌体外生长的影响。两种蜈蚣的2%NaHCO_3和蒸馏水提取液以及油脂对真菌影响极小;3%CH_3COOH 提取液强烈抑制真菌生长,但无杀菌作用,测定了相应的最小抑菌浓度(MIC)。各提取物对细菌均无抑制作用。 展开更多
关键词 少棘蜈蚣 墨江蜈蚣 真菌 细菌
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中药蜈蚣毒溶血活性试验 被引量:15
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作者 邓芬 方红 王克勤 《中药材》 CAS CSCD 1997年第1期36-37,共2页
通过溶血试验对活体少棘蜈蚣,药材少棘蜈蚣和多棘蜈蚣的粗毒进行溶血活性比较。结果表明均有溶血活性,且药材蜈蚣毒活性较活体蜈蚣大大降低,陈药材蜈蚣毒活性较新鲜药材降低一倍,多棘蜈蚣毒活性明显高于少棘蜈蚣。
关键词 溶血活性 毒素 少棘蜈蚣 多棘蜈蚣
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