温郁金二萜类化合物C对人结肠癌细胞SW620增殖的影响

目的观察温郁金醚提物中二萜类化合物C（以下简称化合物C）体外对人结肠癌细胞SW620生长、细胞凋亡和细胞周期的影响。方法以顺铂（DDP）为阳性对照药物,采用噻唑蓝（MTT）法检测化合物C对SW620细胞增殖的抑制作用; An nexin V FITC标记流式细胞术（FCM）检测化合物C对SW620细胞的凋亡抑制率,流式细胞术检测化合物C对SW620细胞周期的影响。结果MTT法显示化合物C对SW620的半数抑制浓度（IC50）为24.16 μg•mL 1,顺铂为45.80 μg•mL 1;化合物C较低浓度时的抑制率与阳性对照组(DDP组)相似,在30,50,70 μg•mL 1时的抑制率与对照组比较,差异有统计学意义（均P＜0.05）;FCM显示,化合物C能诱导SW620细胞凋亡,其诱导凋亡率呈现一定的浓度和时间依赖性,48 h后化合物C70 μg•mL 1组凋亡率为33.55%;化合物C能将SW620细胞阻滞于G1期,减少肿瘤细胞在S期和G2/M的比例。结论化合物C能诱导SW620细胞凋亡,阻滞其细胞周期,抑制细胞增殖,是温郁金发挥抗肿瘤作用的有效成分之一。     

丝裂霉素C联合原花青素对人膀胱癌BIU87细胞的生长抑制作用

［摘要］目的探讨丝裂霉素C（MMC）与葡萄籽提取物原花青素（grape seed procyanidin extract, GSPE）联合应用对人膀胱癌BIU87细胞的生长抑制作用。方法采用噻唑蓝（MTT）比色法测定MMC和GSPE单独和联合应用时对人膀胱癌BIU87细胞生长的抑制作用,并应用Chou Talalay联合指数法定量分析药物之间的相互作用效果。结果MMC、GSPE单独应用时,BIU87细胞生长的抑制作用随药物浓度的增加而增加,中效浓度分别为15.422,195.865 μg•mL 1。MMC与GSPE联合应用时,作用效果表现为在低浓度（0%＜fa＜37%）时呈协同作用（CI＜1）,高浓度（37%＜fa＜100%）时呈拮抗作用（CI＞1）,中效浓度为109.496 μg•mL 1 （其中MMC 8.111 μg•mL 1,GSPE 101.385 μg•mL 1）。结论MMC和GSPE相互作用的效果在低浓度时呈协同作用,高浓度时呈拮抗作用。     

丹参酮化合物对HeLa细胞抑制作用与构效关系探讨

目的 研究丹参酮化合物对HeLa细胞增殖的抑制作用,阐明其结构与细胞毒活性的关联性. 方法 MTT比色法检测不同浓度的丹参酮化合物,分别在24,48,72 h对HeLa细胞增殖的抑制作用. 结果 不同浓度（0.5～16.0 μg•mL^-1）丹参酮ⅡA、丹参酮I、二氢丹参酮、隐丹参酮对HeLa细胞毒性均有明显剂量和时间依赖性,其作用24 h的IC50值分别为：24.1,20.1,8.4,17.8 μg•mL^-1;作用48 h的IC₅₀值分别为：8.0,11.1,2.2,8.2 μg•mL^-1;作用72 h的IC₅₀值分别为：4.6,3.5,1.5,8.1 μg•mL^-1. 结论 丹参酮化合物对HeLa细胞均具有增殖抑制作用,其中二氢丹参酮对宫颈癌细胞系HeLa的细胞毒性最强. A环为芳香环与D环呋喃环的结构差别使其对HeLa细胞增殖的抑制作用有明显的差异.     

地锦草提取物抗真菌作用及对红色毛癣菌角鲨烯环氧化酶的影响

目的 对地锦草提取物（EHE）进行体外抗真菌实验,探讨其抗真菌作用机制。方法 采用美国临床实验室标准化委员会（NCCLS）推荐的《产孢丝状真菌的液基稀释法抗真菌药物敏感性试验方案》（M38 A）,测定地锦草提取物对82株临床常见真菌的最低抑菌浓度值（MIC）;酶联免疫吸附试验（ELISA）研究其对红色毛癣菌角鲨烯环氧化酶的影响。结果 地锦草提取物对红色毛癣菌的平均MIC为446 μg•mL 1,石膏样毛癣菌值的平均MIC为652 μg•mL 1,紫色毛癣菌的平均MIC为1 024 μg•mL 1,许兰毛癣菌的平均MIC为896 μg•mL 1,疣状毛癣菌平均MIC为853 μg•mL 1,犬小孢子菌及4种念珠菌的平均MIC均＞1 024 μg•mL 1;地锦草提取物256 μg•mL 1时,可显著降低红色毛癣菌中角鲨烯环氧化酶的活性（P＜0.01）。结论 地锦草提取物有显著的抗真菌活性,对皮肤癣菌的敏感性比念珠菌高;其抗真菌机制可能与抑制角鲨烯环氧化酶的活性有关。     

黄连提取物诱导HaCat细胞凋亡作用研究

目的探讨黄连提取物对HaCat细胞凋亡及核因子κB（NF κB）基因转录水平的影响。方法以41.6,20.8,10.4 μg•mL－1黄连提取物作用于HaCat细胞,采用噻唑蓝（MTT）法测定HaCat细胞增殖情况,DNA Ladder法检测黄连提取物对HaCat细胞凋亡的影响,半定量逆转录聚合酶链反应（RT-PCR）测定给药后吸光度值,计算各组NF κB /磷酸甘油醛脱氢酶（NF-κB/GAPDH）值,分析NF-κB基因转录水平变化。结果黄连提取物对HaCat细胞增殖具有抑制作用,抑制强度呈浓度依赖性,41.6 μg•mL－1黄连提取物抑制作用最强（抑制率66.17%）。不同剂量黄连提取物导致HaCat细胞呈阶梯状DNA,呈明显凋亡特征。41.6和20.8 μg•mL－1黄连提取物对NF-κB基因转录具有显著下调作用。结论黄连提取物可诱导HaCat细胞凋亡,并下调NF-κB基因转录水平。     

慢性阻塞性肺疾病患者小剂量茶碱的血药浓度监测

［摘要］目的探讨慢性阻塞性肺疾病（COPD）患者小剂量茶碱治疗的合理血药谷浓度,并分析影响茶碱血药浓度的因素。方法对60例接受小剂量茶碱治疗的患者的血药浓度监测结果进行了回顾性分析。结果茶碱血清浓度＜3 μg•mL 1的 2例,3～6 μg•mL 1的53例,肺心病患者茶碱血药浓度偏高; 嗜烟患者茶碱血药浓度偏低。结论小剂量茶碱治疗COPD患者时,建议血清谷浓度调控在3～6 μg•mL 1。吸烟对茶碱血药浓度有明显影响。     

固相萃取离子色谱法测定反义寡核苷酸药物杂质

目的建立同时测定寡核苷酸原料药中杂质醋酸（MCAA）、二氯乙酸（DCAA）、氯离子（Cl－）含量的固相萃取离子色谱（IC）法。方法Ionpac AS 23（4 mm×250 mm）分析柱;Ionpac AG 23（4 mm×50 mm）保护柱;流动相：0.8 mmol•L－1 碳酸氢钠/4.5 mmol•L－1碳酸钠;抑制型电导检测器。结果MCAA、DCAA和Cl－的标准曲线方程分别为：Y＝ 5.230 7X + 0.014 （r＝0.999 7）,Y＝ 19.762X－0.007 2 （r＝0.999 9）和Y＝507.4X－0.746 2 （r＝0.999 8）;MCAA在0.002～0.400 μg•mL－1浓度范围内呈良好的线性关系,DCAA在0.000 624～0.156 μg•mL－1浓度范围内呈良好的线性关系,Cl－在2.4×10－3～1.5 mg•mL－1浓度范围内呈良好的线性关系,其检测限分别为0.02,0.312 ng•mL－1和0.01 μg•mL－1。结论该方法操作简便、快速,结果准确,可用于反义寡核苷酸原料药中杂质的控制。     

反相高效液相色谱法测定人血浆洛拉曲克浓度

目的建立测定人血浆洛拉曲克浓度的反相高效液相色谱法。方法以DiamonsilTM C18反相柱（150 mm×4.6 mm, 5 μm）为色谱柱,流动相为0.03 mol•L 1醋酸铵 甲醇（40：60）;流速：0.8 mL•min 1;柱温：40 ℃;检测波长： 233 nm。以乙酸乙酯与二氯甲烷（80：20）为提取剂。结果洛拉曲克的高、中、低（50.0, 10.0,1.0 μg•mL 1）3种浓度平均回收率分别为98.72%,97.86%,101.27%,日内、日间差RSD均＜7%（n＝5）;分析方法的检测限为1.0 μg•mL 1;线性范围为1.0～50.0 μg•mL 1。标准曲线方程： Y＝1.73X－5.29,r＝0.999 8（n＝8）。结论该方法灵敏、准确、简单、快速,可用于临床血药浓度监测和药动学研究。     

高效液相色谱法测定 妇炎洁栓中大黄素与大黄酚的含量

［摘要］目的建立妇炎洁栓中大黄素和大黄酚的含量测定方法。方法采用高效液相色谱法，流动相：甲醇 0.1%磷酸（78：22）；流速：0.9 mL•min 1；检测波长：254 nm；柱温：40 ℃。结果大黄素和大黄酚分别在1.015～8.120 μg•mL 1和1.325～10.600 μg•mL 1范围内线性关系良好；大黄素的平均回收率为99.3%（RSD=1.20%），大黄酚的平均回收率为99.3%（RSD=1.35%）。结论该方法简便快速，结果准确、可靠，可用于该制剂的质量控制。     

透皮促渗剂对醋酸地塞米松壳聚糖凝胶透皮特性的影响

目的观察透皮促渗剂对醋酸地塞米松壳聚糖凝胶透皮特性的影响。方法实验分为无促渗剂组和促渗剂组。无促渗剂组为0.75%药物5%壳聚糖凝胶剂;促渗剂组根据含促渗剂不同又分为月桂氮酮+丙二醇、月桂氮酮、丙二醇+二甲亚砜、二甲亚砜+月桂氮酮组。以无毛大鼠皮肤为渗透屏障,进行体外渗透实验,分析该凝胶稳态透皮速率（Js）和Js提高率。结果无促渗剂组①Js为（3.75±0.56） μg•（cm2） 1•h 1。促渗剂组效果明显,其中二甲亚砜+月桂氮酮组②Js为（8.12±0.58） μg•（cm2） 1•h 1,月桂氮酮+丙二醇组③Js为（5.41±0.74） μg•（cm2） 1•h 1,丙二醇+二甲亚砜组④Js为（4.31±0.42） μg•（cm2） 1•h 1,月桂氮酮组⑤Js为（4.35±0.36） μg•（cm2） 1•h 1。与①比较,②的Js提高率为2.17%（P＜0.01）,与③④⑤比较,②的Js分别为1.51,1.89,1.87倍（P＜0.05或P＜0.01）。结论混合促渗剂具有比单一促渗剂更好的促渗效果。     

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.     

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.     

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.     

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.