正交试验法优选狼毒大戟中活性成分的提取工艺

摘 要 目的：优选狼毒大戟中活性成分的最佳提取工艺。方法: 选用L9(3⁴ )正交试验设计,以提取物中岩大戟內酯B和狼毒乙素的含量为指标,考察乙醇浓度、溶剂用量、提取温度和提取时间对提取效果的影响。结果:最佳提取工艺为料液比1∶〖KG-*2〗10,加入浓度为70%的乙醇溶液,以70 ℃为提取温度,加热回流提取1 h。结论:优选得到的提取工艺稳定、合理、可行,操作简单,能耗低,大大提高了岩大戟内酯B和狼毒乙素的提取率。     

狼毒大戟配伍大枣合煎液与单煎液主要成分含量的比较研究

目的：建立狼毒大戟与大枣合煎前后提取液中有效成分含量的分析方法,比较2种提取液中岩大戟内酯B和狼毒乙素的含量差异。方法：采用UPLC-DAD方法,对狼毒大戟配伍大枣合煎液与单煎液中的上述2个主要成分进行含量测定。应用Waters ACQUITY BEH C₁₈柱（2.1 mm×100 mm,1.8 μm）,0.1%甲酸水与色谱乙腈进行梯度洗脱,流速为0.8 mL·min^-1,检测波长210 nm,柱温为30℃,进样量为5 μL。结果：这两个化合物在测定的范围内表现出良好的线性（r>0.998 4）,方法的回收率在92.6%~103.4%之间。狼毒大戟配伍大枣合煎液中岩大戟内酯B和狼毒乙素的含量均显著低于单煎液。结论：狼毒大戟与大枣合煎可降低有毒成分的溶出。     

天山假狼毒和瑞香狼毒根中挥发性成分研究

目的用GC-MS法分析比较天山假狼毒和瑞香狼毒根中挥发性成分的异同。方法采用索氏提取法提取天山假狼毒和瑞香狼毒根中的挥发性成分,经GC-MS联用仪对挥发性成分进行分析鉴定,采用色谱峰面积归一化法计算各挥发性成分的含量。结果共鉴定出2种植物根中的113个挥发性成分,其中天山假狼毒81个,瑞香狼毒69个,二者有37个共有成分。相对百分含量较高的成分有:十六酸甲酯(hexadecanoic acid-methyl ester)、9,12-顺式十八碳二烯酸[(Z,Z)-9,12-octadecadienoic acid]、角鲨烯(squalene)、1-庚三醇(1-heptatriacotanol)和异柠檬酸乙酯(ethyl iso-allocholate)。结论天山假狼毒和瑞香狼毒根中的挥发性成分有部分相近,但在含量与其他单一组分组成上存在一定的差异,本研究为天山假狼毒和瑞香狼毒药材的质量控制和进一步研究提供了重要理论依据。     

冬凌草抗炎有效部位筛选及初步表征

目的:筛选冬凌草的抗炎有效部位并对有效部位进行初步表征.方法:冬凌草采用梯度极性溶剂水、25%乙醇、50%乙醇、75%乙醇及100%乙醇分别提取,提取物通过蛋清致大鼠足跖肿胀试验以确定抗炎有效部位;并对有效部位采用MS分析,HPLC测定主成分冬凌草甲素的含量.结果:冬凌草100%乙醇提取物对大鼠足跖肿胀有明显的抑制作用,MS分析提取物中明显含有冬凌草甲素及其他贝壳杉烷类二萜,HPLC测定无水乙醇提取冬凌草甲素的提取率为0.174 6%.结论:冬凌草100%乙醇提取物组分具有明显的抗炎作用,为冬凌草的抗炎有效部位,为其临床抗炎提供依据.     

不同极性溶媒瑞香狼毒提取物的抗肿瘤活性比较

目的：通过比较不同极性溶媒瑞香狼毒提取物的体外抗肿瘤活性，确定瑞香狼毒抗肿瘤活性成分所在的极性区间。方法：采用索氏回流提取法，依次用石油醚、乙酸乙酯、丙酮和乙醇对狼毒进行回流提取，并用不同浓度的各溶剂提取物分别处理Eca109、K562、HepG_2细胞株，通过MTT检测法，比较各溶媒提取物对肿瘤细胞的抑制活性。结果：石油醚提取物和乙酸乙酯提取物对体外培养肿瘤细胞表现出较强的抑制作用，在测定浓度范围内呈现出良好剂量依赖性，石油醚提取物最大抑制率分别为78％(Eca-109)、93％(K562)、95％(HepG_2)，乙酸乙酯提取物最大抑制率分别为53％(Eca-109)、91％(K562)、87％(HepG_2)；而丙酮和乙醇提取部位在测定浓度范围内则显示出很弱的抑制作用；在各提取物中石油醚提取物的体外抗肿瘤作用最强。结论：石油醚提取成分是瑞香狼毒中体外抗肿瘤活性最强的部位。     

白狼毒化学成分和生物活性研究进展

白狼毒作为有毒中药,具有抗结核、抗癌、抗白血病、细胞毒、免疫调节等活性.本文综述了近10年来狼毒大戟和月腺大戟中80多个成分的化学研究,为白狼毒临床应用的安全有效和进一步深入研究提供参考.     

狼毒的有效成分及其药理活性研究进展

狼毒是毒属植物瑞香科瑞香狼毒 (Stellera chamaejasmeL.)或大戟科狼毒大戟 (Euphorbia fischeriana Stued.)、月腺大戟 (Euphorbia ebracteolata Hayata.)的根 ,有逐水祛痰、散结杀虫之功效 ,为常用中药。近年的研究发现 ,狼毒提取物对肿瘤细胞有较强的抑制活性 ,此外还有抗菌和抗病毒作用 ,是一个理想的天然药物开发品种。1 　 化学成分研究1 .1　瑞香狼毒　六十年代初 ,我国的研究者从瑞香狼毒的根中分离出一种酸性物质 ,定为狼毒素 (chamaejamine) ,后来确定了其双氢双黄酮的结构。目前瑞香狼毒中已知的成分主要有倍半萜内酯、二萜生…     

红花中酪氨酸酶抑制成分的快速筛选和分析

目的 通过红花中酪氨酸酶抑制剂的筛选,明确红花中酪氨酸酶抑制剂成分,为美白祛斑剂的研发提供实验依据.方法 采用索氏提取法依次提取红花石油醚、氯仿、醋酸乙酯、正丁醇和水5种提取物,分别检测其对酪氨酸酶的抑制作用;针对活性组分通过UPLC-Q/TOF鉴定活性成分,并结合分子对接和酶学检测验证药效成分.结果 初步筛选结果显示红花水层提取物具有较强的抑制作用,并进一步通过液相色谱分离,质谱检测,以及分子对接预测技术,推测其中9种成分可能具有酪氨酸酶抑制作用,选取代表性单体羟基红花黄色素A进行验证,确认其具有较强的抑制作用.结论 谱效结合构效的筛选模式为天然产物中美白祛斑成分的筛选提供了便捷方法;红花中羟基红花黄色素A对酪氨酸酶具有明显的抑制作用.     

狼毒大戟中松香烷型二萜内酯的抗肿瘤活性研究

摘 要 目的： 对狼毒大戟中松香烷型二萜内酯类化学成分进行分离和结构鉴定,将得到的单体化合物与6种不同的人肿瘤细胞株相互作用探讨其抗肿瘤活性。方法: 通过硅胶、ODS等柱色谱与1D、2D-NMR波谱学测试技术结合,从狼毒大戟中分离并确定4个松香烷型二萜内酯类化合物：jolkinolide A (1)、jolkinolide B (2)、17 hydroxyjolkinolide A (3)、17-hydroxyjolkinolide B (4)。采用噻唑蓝比色法（MTT法）分别测定其与6种人肿瘤细胞株：肝癌HepG-2、乳腺癌MCF-7、胃癌SGC 7901、胃癌BGC-823、胃癌MGC-803、宫颈癌Hela相互作用结果,并计算抑制率与半抑制率(IC₅₀)。结果: 化合物2对肝癌HepG-2、乳腺癌MCF-7、宫颈癌Hela细胞的增殖有显著得抑制作用IC₅₀均＜25 μmol·L^-1,化合物4对乳腺癌MCF 7细胞的增殖有显著抑制作用IC₅₀＜30 μmol·L^-1。结论: 狼毒大戟中松香烷型二萜内酯类化合物具有显著的抗肿瘤活性。     

液-质联用鉴别中药狼毒药材的来源

目的建立液-质联用鉴别中药狼毒药材来源的方法。方法瑞香狼毒、狼毒大戟及月腺大戟药材饮片分别粉碎后用70%乙醇超声提取30 min,提取液经微孔滤膜过滤后,利用液-质联用法进行分析。结果从3种狼毒药材提取物的液相色谱及质谱图中选择18个紫外吸收及质谱响应较好的色谱峰,并对其定性,可作为不同种属狼毒药材鉴别的特征峰。结论该方法简便快速,能够用于狼毒药材饮片来源的鉴别分析。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.