浅谈《中国药典》药材与饮片毒性的非临床研究与评价

通过对中药中毒原因的浅析及对《中国药典》药材及饮片毒性分类存在缺陷的分析，简要论述了《中国药典》药材与饮片毒性的非临床研究与评价的必要性，并对其非临床研究与评价的内容等进行了探讨，指出《中国药典》药材与饮片毒性的非临床研究与评价对科学制定毒性分级标准、规范管理使用药材及饮片具有重要意义。     

Changes in Growth,Photosynthesis Performance,Pigments, and Toxin Contents of Bloom-Forming Cyanobacteria after Exposure to Macroalgal Allelochemicals

Macroalgae can directly restrict the growth of various phytoplankton species by releasing allelopathic compounds; therefore, considerable attention should be paid to the allelopathic potential of these organisms against harmful and bloom-forming cyanobacteria. The main aim of this study was to demonstrate for the first time the allelopathic activity of Ulva intestinalis on the growth, the fluorescence parameters: the maximum PSII quantum efficiency (F_v/F_m) and the effective quantum yield of PSII photochemistry (ΦPSII), the chlorophyll a (Chl a) and carotenoid (Car) content, and the microcystin-LR (MC-LR) and phenol content of three bloom-forming cyanobacteria, Aphanizomenon sp., Nodularia spumigena, and Nostoc sp. We found both negative and positive allelopathic effects of U. intestinalis on tested cyanobacteria. The study clearly showed that the addition of the filtrate of U. intestinalis significantly inhibited growth, decreased pigment content and F_v/F_m and ΦPSII values of N. spumigena and Nostoc sp., and stimulated Aphanizomenon sp. The addition of different concentrations of aqueous extract also stimulated the cyanobacterial growth. It was also shown that the addition of extract obtained from U. intestinalis caused a significant decrease in the MC-LR content in Nostoc sp. cells. Moreover, it the phenol content in N. spumigena cells was increased. On the other hand, the cell-specific phenol content for Aphanizomenon sp. decreased due to the addition of the filtrate. In this work, we demonstrated that the allelopathic effect of U. intestinalis depends on the target species’ identity as well as the type of allelopathic method used. The study of the allelopathic Baltic macroalgae may help to identify their possible role as a significant biological factor influencing harmful cyanobacterial blooms in brackish ecosystems.     

间苯三酚与利多卡因联用对绝经期妇女取环术效果的评价

目的：评价间苯三酚与利多卡因联用对绝经后妇女宫内节育器取出术的使用效果。方法：将绝经后行取环术的87例妇女分为观察组和对照组,观察组予术前间苯三酚联合术中局部用利多卡因阻滞麻醉,对照组患者予术中常规使用利多卡因局部阻滞麻醉,比较2组患者术中宫颈口松弛情况和镇痛及取环效果。结果：观察组患者的镇痛和取环效果显著优于对照组（P〈0.05）;前者宫颈口松弛情况与后者比较,其差异无统计学意义（P〉0.05）。结论：间苯三酚与利多卡因用于绝经妇女取环术,可显著减轻患者疼痛,改善取环效果。     

三维重组眼角膜上皮模型法与兔眼刺激法对洗发水眼刺激性的比较与评价

目的 本文同时采用三维人重组角膜模型BioOcular?和现行的兔眼刺激Daize法,对市面常见的不同配方洗发水的眼刺激性进行评价与比较。方法 参考体外眼刺激实验(OECD指导原则492),采用人重组眼角膜上皮模型和兔眼刺激法对8种不同品牌洗发水的眼刺激性进行评价研究。结果 研究表明,与传统兔眼实验比较,采用皮肤模型的眼刺激法对8种洗发水有无刺激性的区分结果与兔眼刺激法基本一致。其中两种方法相关系数r达到0.955,具有很好的相关性。结论 与兔眼刺激法相比,人重组眼角膜上皮模型刺激实验所需的周期短、试验步骤相对简便、检测结果相对更加客观,人源重组组织与人眼刺激更为接近,是一种值得推荐的眼刺激替代检测方法。     

Toxicity of Lithium to Three Freshwater Organisms and the Antagonistic Effect of Sodium

Lithium (Li) is the lightest metal and occurs primarily in stable minerals and salts. Concentrations of Li in surface water are typically <0.04mg l^–1 but can be elevated in contaminated streams. Because of the general lack of information concerning the toxicity of Li to common toxicity test organisms, we evaluated the toxicity of Li to Pimephales promelas (fathead minnow), Ceriodaphnia dubia, and a freshwater snail (Elimia clavaeformis). In the laboratory, the concentration of Li that inhibited P. promelas growth or C. dubia reproduction by 25% (IC25) was dependant upon the dilution water. In laboratory control water containing little sodium (2.8mg l^–1), the IC25s were 0.38 and 0.32mg Li l^–1 and in ambient stream water containing 17mg Na l^–1, the IC25s were 1.99 and 3.33, respectively. A Li concentration of 0.15mgl^–1 inhibited the feeding of E. clavaeformis in laboratory tests. Toxicity tests conducted to evaluate the effect of sodium on the toxicity of Li were conducted with fathead minnows and C. dubia. The presence of sodium greatly affected the toxicity of Li. Fathead minnows and Ceriodaphnia, for example, tolerated concentrations of Li as great as 6mg l^–1 when sufficient Na was present. The interaction of Li and Na on the reproduction of Ceriodaphnia was investigated in depth and can be described using an exponential model. The model predicts that C. dubia reproduction would not be affected when animals are exposed to combinations of lithium and sodium with a log ratio of mmol Na to mmol Li equal to at least 1.63. The results of this study indicate that for most natural waters, the presence of sodium is sufficient to prevent Li toxicity. However, in areas of historical disposal or heavy processing or use, an evaluation of Li from a water quality perspective would be warranted.     

两种方法测定比沙可啶纯度及不确定度评定

目的:采用两种不同原理的测定方法对比沙可啶纯度进行联合定值,并建立其不确定度评定方法。方法:采用高效液相色谱法(HPLC)与非水溶液电位滴定法测定比沙可啶纯度,并依据相关规范要求,对两种方法测定过程的不确定度进行了系统分析。结果:比沙可啶HPLC法、非水溶液电位滴定法纯度测定值分别为(99.81±0.29)%、(99.82±0.51)%,两种方法联合测定比沙可啶纯度的标准值及其不确定度分别为99.82%、(±)0.59%(k=2,P=0.95)。结论:采用HPLC与非水溶液电位滴定法联合测定比沙可啶纯度及不确定度评定结果准确可靠,避免了采用一种技术带来的分析方法缺陷,有利于提高比沙可啶的质量评价与控制水平,同时为比沙可啶纯度标准物质的研制提供了科学依据。     

生物技术药物的发育和生殖毒性评价

由于生物技术药物与小分子药物本身存在差异,发育和生殖毒性(DART)评价应考虑受试物的种属特异性、免疫原性、生物活性和暴露情况,并采取灵活、个案处理和基于科学的方法进行研究.当传统种属(如啮齿类、兔)不是药理学相关物种时,DART评价首选非人灵长类动物模型.若生物技术药物只与人或黑猩猩产生交叉反应,DART评价可使用针对传统种属开发的同源蛋白(替代分子)作为受试物.此外,还可使用转基因动物进行DART评价.     

基于斑马鱼整合评价补骨脂粗多糖对补骨脂中香豆素组分的代谢及毒效关系的影响

目的 用斑马鱼整合评价探讨补骨脂粗多糖(psoraleae crude polysaccharide,PPS)对补骨脂中香豆素组分(coumarin components of Psoraleae Fructus,CCPF)的代谢及毒效关系影响。方法 用受精后天数(days post fertilization,dpf)1~6 dpf的斑马鱼评价CCPF、PPS及二者组合物的安全性,观察斑马鱼脏器形态,记录死亡数并计算斑马鱼半数死亡浓度(LC₅₀);用1~6 dpf斑马鱼暴露于CCPF及其与PPS组合物,分析补骨脂苷和异补骨脂苷及其代谢物补骨脂素和异补骨脂素的动态变化;用25 μmol·L^-1泼尼松龙诱导斑马鱼骨质疏松模型,采用茜素红对培养至8 dpf的各给药组斑马鱼幼鱼骨骼染色,进行显微观察、数码成像,并用图像软件定量分析骨骼染色区域来评价上述样品的抗骨质疏松活性。结果 CCPF与PPS组合物对斑马鱼的毒性大于二者单独给药,且组合物PPS的比例增加,毒性增大：PPS能够降低斑马鱼中毒浓度,致斑马鱼脏器形态畸变严重,缩短斑马鱼死亡时间,增加斑马鱼死亡率;PPS能明显加速CCPF中补骨脂苷和异补骨脂苷脱糖基转化为潜在肝损伤代谢产物补骨脂素和异补骨脂素;CCPF及其与PPS组合物可增加斑马鱼头骨累积光密度值,且组合物具有一定增效作用。结论 用斑马鱼整合评价高效揭示了PPS对CCPF的代谢与毒效关系的影响,为研究PPS对其他结构类型成分的毒效关系提供高效的方法与思路。     

In Vitro and In Vivo Evaluation of Amorphous Solid Dispersions Generated by Different Bench-Scale Processes, Using Griseofulvin as a Model Compound

Drug polymer-based amorphous solid dispersions (ASD) are widely used in the pharmaceutical industry to improve bioavailability for poorly water-soluble compounds. Spray-drying is the most common process involved in the manufacturing of ASD material. However, spray-drying involves a high investment of material quantity and time. Lower investment manufacturing processes such as fast evaporation and freeze-drying (lyophilization) have been developed to manufacture ASD at the bench level. The general belief is that the overall performance of ASD material is thermodynamically driven and should be independent of the manufacturing process. However, no formal comparison has been made to assess the in vivo performance of material generated by different processes. This study compares the in vitro and in vivo properties of ASD material generated by fast evaporation, lyophilization, and spray-drying methods using griseofulvin as a model compound and hydroxypropyl methylcellulose acetate succinate as the polymer matrix. Our data suggest that despite minor differences in the formulation release properties and stability of the ASD materials, the overall exposure is comparable between the three manufacturing processes under the conditions examined. These results suggest that fast evaporation and lyophilization may be suitable to generate ASD material for oral evaluation. However, caution should be exercised since the general applicability of the present findings will need to be further evaluated.     

应用Markov模型对甘精胰岛素和利拉鲁肽不良事件的药物经济学评价

摘 要 目的：针对甘精胰岛素（Glar）和利拉鲁肽（Lira）的主要不良事件进行长期经济学评价,为2型糖尿病患者药物选择提供参考依据,以期在可承受的经济水平内提高糖尿病患者的生命质量。方法：根据研究药物的主要不良事件建立Markov模型,收集状态间的转移概率、各状态成本及效用值等数据,借助TreeAge Pro 2011软件对该模型进行回乘分析、半循环矫正及敏感性分析,并计算增量成本效果比（ICER）。结果：Glar、Lira 1.2mg及Lira 1.8 mg治疗20年的人均成本分别为70 948.70,93 939.31,138 108.92元,分别获得5.14,6.84,7.16个质量调整生命年（QALYs）。与Glar相比,Lira 1.2mg及Lira 1.8mg的增量成本效果比（ICER）分别为13 523.89,33 247.63。结论：与Glar相比,Lira两种干预措施均具有成本 效果优势,且Lira 1.2mg的经济性更好。     

Evaluation of Disintegration Testing of Different Fast Dissolving Tablets Using the Texture Analyzer

The in vitro disintegration behavior of fast dissolving systems manufactured by the main commercialized technologies was studied using the texture analyzer (TA) instrument. Quantitative parameters were employed to characterize the effect of the major test variables on the disintegration profiles. The average disintegration profiles of the products were compared using the test conditions that minimized these effects and at the same time mimicked the in vivo situation in the patient's mouth. The differences in the disintegration mechanisms of the fast dissolving systems were reflected in the shape of their disintegration profiles and in the parameters derived from the profiles. The differences were explained in relation to the technology and/or formulation characteristics involved in the manufacture of each product. The in vitro disintegration times obtained under the simulated in vivo conditions were correlated with the reported in vivo disintegration times.     

Phytotoxicity Evaluation of Type B Trichothecenes Using a Chlamydomonas reinhardtii Model System

Type B trichothecenes, which consist of deoxynivalenol (DON) and nivalenol (NIV) as the major end products, are produced by phytotoxic fungi, such as the Fusarium species, and pollute arable fields across the world. The DON toxicity has been investigated using various types of cell systems or animal bioassays. The evaluation of NIV toxicity, however, has been relatively restricted because of its lower level compared with DON. In this study, the Chlamydomonas reinhardtii testing system, which has been reported to have adequate NIV sensitivity, was reinvestigated under different mycotoxin concentrations and light conditions. The best concentration of DON and NIV, and their derivatives, for test conditions was found to be 25 ppm (2.5 × 10⁻² mg/mL). In all light test conditions, DON, NIV, and fusarenon-X (FusX) indicated significant growth inhibition regardless of whether a light source existed, or under differential wavelength conditions. FusX growth was also influenced by changes in photon flux density. These results suggest that C. reinhardtii is an appropriate evaluation system for type B trichothecenes.     

前列地尔冻干乳的溶血性和急性毒性评价

本课题对前列地尔冻干乳剂进行了溶血性和急性毒性的安全性考察。通过对兔的溶血性试验说明在所采用的试验条件下注射用前列地尔冻干乳静注不会引起溶血反应;同时试验表明本品具备足够的安全性。以上试验说明本工艺处方所得前列地尔冻干乳生物相容性较好,供静脉注射给药安全。     

甘遂不同炮制品及提取物对斑马鱼的急性毒性研究

目的以模式生物斑马鱼为实验对象,评价甘遂不同炮制品及提取方法的急性毒性。方法采用回流提取方法制备甘遂不同炮制品的水提液和醇提液;将它们的提取液按几何级数设置浓度梯度,添加到鱼生活的水中,观察给药后96 h鱼只的死亡情况,以此为判断待测药物毒性大小的依据,采用SPSS Statistics 17软件计算不同炮制品水提液和醇提液对斑马鱼的半数致死浓度(LC50)。结果斑马鱼对甘遂不同炮制品的水提液和醇提液均表现出急性毒性反应,且毒性作用呈现出明显的量-毒关系;不同炮制品水提液LC50明显高于相应醇提液;同一提取方法不同炮制品的急性毒性大小顺序为甘遂生品>清炒品>醋润品>醋炙品。结论以斑马鱼作为实验动物,甘遂生品的醇提液急性毒性最强、甘遂醋炙品的水提液急性毒性最低。本实验为进一步认识与评价甘遂毒性及醋炙减毒机制提供了依据。     

中药毒性评价的技术方法与应用

中药的安全性近年来受到了广泛的关注和重视,但是相应的理论和实验研究还不够充分。中药的毒性是中药安全性的关键问题之一,参照国际惯例和西方植物药研究的标准规范,依据中医药理论,应用现代新技术新方法,结合基础研究的成果,制定相应的评价体系和研究规范,是解决这个关键问题的可行思路。对近年来课题组按照上述研究思路开展的有关中药毒性评价的技术方法与应用研究的结果进行了概述,希望能为中药毒性的评价方法和技术指导原则提供理论基础,也可为雷公藤等典型中毒中药的毒性研究提供研究示范,推动中药毒性研究的发展。     

基于斑马鱼毒/效同步评价的壮骨关节丸促骨骼发育作用与初步安全性研究

目的 用斑马鱼毒/效同步法评价壮骨关节丸的促骨骼发育作用及初步安全性。方法 将受精后3 d（3 day post fertilization,3 dpf）的斑马鱼幼鱼暴露于25 μmol·L^-1泼尼松龙（prednisolone,PN）,壮骨关节丸溶液及含PN的壮骨关节丸溶液（12.5,25,50,100,200,300和400 μg·mL^-1）中,0.5% DMSO胚胎培养基为空白对照,于28℃培养至8 dpf,隔天换一半药液。采用茜素红对8 dpf的斑马鱼幼鱼头部骨骼染色,定量分析骨骼染色区域;给药期间每天同步观察鱼死亡及脏器中毒情况。结果 在8 dpf时,100 μg·mL^-1及以上浓度的壮骨关节丸健康鱼组和含PN的壮骨关节丸模型鱼组斑马鱼死亡率>80%,未进行骨骼染色。壮骨关节丸健康鱼组（12.5,25,50 μg·mL^-1）的斑马鱼头骨矿化面积和累积光密度值IOD与PN组比较显著增加（P<0.01或<0.05）,而含PN的壮骨关节丸模型鱼组（12.5,25,50 μg·mL^-1）斑马鱼的头骨矿化面积或IOD结果显示阻止PN致骨丢失的趋势,但均低于健康鱼组。给予壮骨关节丸后,PN模型鱼的死亡速度和死亡率较健康鱼高,鱼死亡率与给药浓度和时间呈依赖性;PN模型鱼的6 dpf和8 dpf半数致死浓度值（63.92,63.92 μg·mL^-1）均低于健康鱼（94.66,87.84 μg·mL^-1）。结论 斑马鱼毒/效同步法快速评价了壮骨关节丸对健康鱼和模型鱼的骨骼发育作用及初步安全性。壮骨关节丸对健康鱼和模型鱼的作用及毒性有差别。     

30种药材两种提取方法提取液微量元素测定和分析

本文采用原子吸收分光光度法测定30种药材、两种提取方法提取液中6种微量元素含量，结果表明6种微量元素在提取过程中WE法有不同程度的丧失、WAE法极大部分丧失。     

Comparison of Cell Proliferation and Toxicity Assays Using Two Cationic Liposomes

The present study compares different cytotoxicity and cell proliferation assays including cell morphology, mitochondrial activity, DNA synthesis, and cell viability and toxicity assays. CaSki cells were exposed to two cationic liposomal preparations containing dimethyldioctadecyl-ammonium bromide (DDAB), dioleoylphosphatidylethanolamine (DOPE) and a commercial transfection-reagent DOTAP(N[l-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium-methylsulfate). The results provided by these assays were similar. However, the lactate dehydrogenase assay was more sensitive in measuring early damages of cell membranes than the Trypan blue assay. Also, cell morphology showed early toxic changes, such as cytoplasmic vacuolization and cell shrinking, and it should be included with such toxicity evaluations. DDAB:DOPE was more toxic than DOTAP. The cells treated with DOTAP at 10 µM were surviving as well as the control cells, while DOTAP at 40 µM and DDAB: DOPE at 10 µM had slight toxic effects on CaSki cells. The most toxic effects were seen in CaSki cells after treatment with DDAB: DOPE at 40 µM.     

清醒大鼠重度失血性休克模型指标建立及有效性评价

目的:制作清醒大鼠重度失血性休克模型,评价模型的有效性,观察不同复苏液对重度失血性休克的疗效。方法:将37只SD大鼠在清醒状态下经股动脉释放其全身总血量的65%,制作重度失血性休克模型,随机分为3组,NaCl组(n=12)、全血组(n=12)和万汶组(羟乙基淀粉130/0.4氯化钠注射液,n=13),分别于放血前、放血末及输液后测定大鼠的平均动脉压(MBP)、呼吸频率(R)、体温(T)、动脉血气以及血乳酸盐(LD)。记录输液后24h与48h的存活率。结果:与放血前比较,各组大鼠放血末MBP、T、pH、二氧化碳分压[p(CO2)]、红细胞比容(HCT)及剩余碱(BE)明显降低,血氧分压[p(O2)]、血乳酸盐水平及氧饱和度升高。输液后2h与NaCl组相比,全血组和万汶组可以明显地改善大鼠的MBP、pH、p(CO2)及BE,降低动脉血乳酸,改善代谢性酸中毒,但全血组恢复效果更明显。结论:清醒大鼠重度失血性休克模型具有满意的重现性和有效性,适于不同复苏液抗休克性能的评价。     

Establishment and Effective Evaluation of Different Indexes of Severe Hemorrhagic Shock Model in Unrestrained Conscious Rats

目的:制作清醒大鼠重度失血性休克模型,评价模型的有效性,观察不同复苏液对重度失血性休克的疗效.方法:将37只SD大鼠在清醒状态下经股动脉释放其全身总血量的65％,制作重度失血性休克模型,随机分为3组,NaCl组(n=12)、全血组(n=12)和万汶组(羟乙基淀粉130/0.4氯化钠注射液,n=13),分别于放血前、放血末及输液后测定大鼠的平均动脉压(MBP)、呼吸频率(R)、体温(T)、动脉血气以及血乳酸盐(LD).记录输液后24h与48h的存活率.结果:与放血前比较,各组大鼠放血末MBP、T、pH、二氧化碳分压[P(CO2)]、红细胞比容(HCT)及剩余碱(BE)明显降低,血氧分压[p(O2)]、血乳酸盐水平及氧饱和度升高.输液后2h与NaCl组相比,全血组和万汶组可以明显地改善大鼠的MBP、pH、p(CO2)及BE,降低动脉血乳酸,改善代谢性酸中毒,但全血组恢复效果更明显.结论:清醒大鼠重度失血性休克模型具有满意的重现性和有效性,适于不同复苏液抗休克性能的评价.