高效液相色谱结合Box-Behnken响应面法优选新疆鼠尾草根萜类成分提取纯化工艺

目的:优化新疆鼠尾草根萜类成分的提取纯化工艺。方法:建立能同时测定新疆鼠尾草根中6,7-去氢罗列酮、齐墩果酸和熊果酸3个萜类成分质量分数的HPLC方法，以3个成分的质量分数为指标，在单因素试验基础上，用Box-Behnken响应面法设计分析分别考察提取温度、提取时间和料液比3个因素对萜类成分提取工艺的影响；考察萃取溶剂比例、萃取次数和振摇时间3个因素对萜类成分纯化工艺的影响。结果:最佳提取工艺为加20倍95%乙醇，85℃回流提取3次，每次2.5 h,在此条件下测得3个成分质量分数之和为4.13%;最佳纯化工艺为向提取物的水混悬液中加等比例的乙酸乙酯萃取5次，每次振摇24 h,在此条件下测得3个成分质量分数之和为21.53%,是提取物的5.19倍。结论:响应面法优选新疆鼠尾草根中萜类成分的提取纯化工艺合理，可为制备新疆鼠尾草根萜类成分提供参考，也为进一步开发利用新疆鼠尾草提供科学依据。     

新二萜醌去氢丹参新酮

从鼠尾草属植物红根草(Salvia prionitis Hance)的根中分到四个二萜醌类化合物,经光谱分析推定为去氢丹参新酮(Ⅰ),丹参新酮(Ⅱ),丹参新醌乙(Ⅲ)和丹参新醌丁(Ⅳ)。Ⅰ为新化合物,所有化合物均系从该植物中首次分到。     

HPLC法同时测定不同产地甘西鼠尾草及丹参中8种主要成分的含量

目的建立同时测定甘西鼠尾草及丹参中丹参素钠、迷迭香酸、紫草酸、丹酚酸B、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA含量的HPLC法,对比不同产地甘西鼠尾草及丹参中8种成分的含量。方法采用SunFire C_(18)(250 mm×4.6 mm,5μm)色谱柱。流动相为乙腈(A)-0.8 mL·L~(-1)甲酸超纯水溶液(B),梯度洗脱(0～19 min,20%A～60%A;19～49 min,60%A～81%A;49～53 min,81%A～91%A;53～57 min,95%A)。流速:1.0 mL·min~(-1);检测波长:270 nm;柱温:26℃;进样量:2μL。结果质量浓度与峰面积线性关系良好(r>0.971 3),精密度、重复性及回收率均符合要求。在50 min内甘西鼠尾草及丹参中8种成分均可实现完全分离。结论青海不同产地甘西鼠尾草较其他产地甘西鼠尾草在脂溶性成分和水溶性成分上均有较好的优势,与丹参相比主要的差别在于丹酚酸B和迷迭香酸的含量,丹酚酸B的含量较丹参样品低,其余4种脂溶性成分均优于丹参样品。     

一种新的HeLa细胞细胞毒剂

一种新的异二氢吲哚酮衍生物的结构为4-羟基-6-甲氧基-1-苯乙基-异二氢吲哚酮,是将猴头菌Hericium erinaceum的子实体用含水有机溶媒(如80％～85％甲醇、乙醇,85％丙酮)室温回流提取1～72h。在40～45C减压蒸除溶媒。得到的水相在有机溶媒(氯仿、乙酸乙酯、乙醚)和水问分配,浓缩有机相得到粗品,再经色谱法和重结晶法得到纯品。该化合物用作HeLa细胞的细胞毒剂。     

去氢丹参新酮对神经炎症的抑制作用及机制研究

目的研究去氢丹参新酮对脂多糖(lipopolysaccharide,LPS)诱导小胶质细胞系BV2细胞产生炎症反应的抑制作用及其作用机制。方法不同浓度去氢丹参新酮预孵育BV2细胞后,用LPS刺激引起神经炎症相关反应。Griess试剂法检测去氢丹参新酮对活化的BV2细胞产生一氧化氮(nitric oxide,NO)的影响,ELISA检测细胞上清液中TNF-α和IL-6的释放量,Confocal观察小胶质细胞表面活化标志物MAC-1表达量的变化,Western blot检测炎症相关信号通路蛋白表达的变化。结果去氢丹参新酮可明显抑制LPS刺激BV2细胞产生的炎症因子包括NO、TNF-α和IL-6的水平,同时抑制一氧化氮合酶、环氧合酶-2等炎症相关蛋白的表达和小胶质细胞表面活化标志物MAC-1的表达。机制研究发现,去氢丹参新酮对PI3K/Akt的过度磷酸化以及NF-κB的过度活化都有明显的抑制作用。结论去氢丹参新酮具有很好的抑制神经炎症活性,其作用机制可能是通过PI3K/Akt信号通路抑制NF-κB的活化而实现的。     

HPLC测定活血止痛丹中芍药苷的含量

活血止痛丹为医院制剂 ,主要由赤芍、丹参、元胡、当归、三七等十余味中药组成 ,具有活血祛瘀、通经活络、消肿止痛的作用 ,主要用于跌打损伤、闪腰岔气等症。处方中干扰成分较多 ,本试验采用将样品置于索氏提取器中 ,加乙醚回流除去干扰成分 ,再以甲醇回流提取 ,用ＨＰＬＣ法测定芍药苷的含量。本法简便 ,重复性好 ,空白无干扰 ,可有效地控制活血止痛丹的质量。1 仪器与试剂高效液相色谱仪 :ＳＰ880 0 (美国 ) ;ＳＰ4 2 90数据处理机 (美国 ) ;ＳＰ84 5 0紫外可见检测器 (美国 )。芍药苷对照品 (中国药品生物制品检定所提供 )。甲醇为色谱…     

新疆鼠尾草中酚酸类化学成分的研究

目的 研究新疆鼠尾草的酚酸类化学成分.方法 采用多种柱色谱技术对化合物进行分离纯化,根据理化数据及波谱数据鉴定化合物的结构.结果 从新疆鼠尾草中分离得到7个化合物,分别鉴定为:丹参素(Ⅰ)、丹参素甲酯(Ⅱ)、原儿茶醛(Ⅲ)、原儿茶酸(Ⅳ)、咖啡酸(Ⅴ)、迷迭香酸(Ⅵ)、丹酚酸K(Ⅶ).结论 化合物Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ为首次从新疆鼠尾草中分离得到.     

丹参中水溶性酚酸类成分的薄层扫描测定法

应用高效薄层扫描法对丹参根中的原儿茶醛、咖啡酸、迷迭香酸及其甲酯、丹酚酸A,B和C7种有效成分进行了含量测定。选择了最佳提取分离方法,即生药用热水提取并酸化后用乙酸乙酯萃取,萃取液浓缩后进行薄层色谱分离。展开剂A:氯仿乙酸乙酯——苯—甲酸(2.4:2:1:0.6)分离原儿茶醛,展开剂B:氯仿-乙酸乙酯-苯-甲酸甲醇(1.5:2:1;1:0.1)分离其余6种成分。并用所建立的分析方法分析比较了4种丹参根中7种有效成分的含量。     

小花清风藤提取齐墩果酸工艺研究

目的优选药材中齐墩果酸成分的最佳提取条件。方法采用正交设计安排试验，选择适当的因素和水平，根据所测含量数据进行统计分析，确定最佳提取条件。结果优选出齐墩果酸的最佳提取工艺条件为小花清风藤药材粉末2g，加入70％的乙醇溶液80mL，水浴回流提取2次，每次1h，抽滤，合并滤液，离心后取上清液浓缩至40mL，加入15％盐酸溶液40mL，加热回流提取4h，再加1倍量水，挥去部分乙醇，用氯仿萃取4次，每次10mL，合并滤液，浓缩至干。结论该提取工艺设计合理，可作为齐墩果酸成分的生产工艺。     

超临界CO2萃取川东獐牙菜总员(口山)酮的工艺优化

目的 优化超临界CO2萃取川东獐牙菜总(口山)酮的工艺.方法 以去甲基雏菊叶龙胆酮和总(口山)酮含量为指标,用正交试验法优选.结果 川东獐牙菜总山酮萃取的最佳工艺条件为:萃取温度40 ℃,萃取压力30 MPa,萃取时间80 min,夹带剂为95%乙醇.结论 采用超临界CO2萃取法提取脂溶性成分具有速度快、效率高和无污染的特点,工艺简便易行.     

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.     

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

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.