Chemical Constituents of Stellera chamaejasme

从瑞香狼毒（ＳｔｅｌｅｒａｃｈａｍａｅｊａｓｍｅＬ）的根中得到１２个化合物。经理化鉴别和光谱分析推定晶（８）为７甲氧基西瑞香素（７ｍｅｔｈｏｘｙｄａｐｈｎｏｒｅｔｉｎ）是新化合物，命名为异西瑞香素（ｉｓｏｄａｐｈｎｏｒｅｔｉｎ），晶（７）为西瑞香素（ｄａｐｈｎｏｒｅｔｉｎ），晶（２）为伞形花内酯（ｕｍｂｅｌｉｆｅｒｏｎｅ），晶（５）为胡萝卜甙，晶（３）为β谷甾醇，晶２，５和８是首次从该植物中分得。     

芫花根中新双香豆素的分离与鉴定

目的研究芫花根(Daphne genkwa)的化学成分。方法采用硅胶和Sephadex LH-20柱色谱技术进行分离纯化，运用多种波谱分析进行结构鉴定。结果分离得到一个新的双香豆素异西瑞香素，结构被确认为7-羟基-6-甲氧基-4［(2-氧杂-2H-1-苯并吡喃-7-基)-氧基］-2H-1-苯并吡喃-2-酮。结论异西瑞香素(isodaphnoretin)为一个新化合物。     

不同产地芫花根皮药材中西瑞香素含量比较

目的采用高效液相色谱（HPLC）法测定不同产地芫花根皮中西瑞香素含量,确定何种产地芫花根皮中西瑞香素含量最高。方法采用Prodigy 5u 100A C18柱（250 mm×4.60 mm,5μm）,流动相为甲醇-0.2%磷酸水溶液（55∶45）,流速为1.0 m L/min,柱温30℃,紫外检测波长346 nm,进样量20μL。结果西瑞香素质量浓度在0.5-20μg/m L范围内与峰面积具有良好的线性关系（r=0.998 4）,平均回收率为98.62%,RSD=4.08%（n=6）。不同产地芫花根皮药材中西瑞香素含量差异较大。结论该法可准确地对芫花根皮药材中有效成分西瑞香素进行定量定性测定,对于保证其内在质量和临床疗效具有重要的意义。     

西瑞香素纳米混悬剂的制备及其体外抗肿瘤作用研究

目的制备西瑞香素纳米混悬剂,并考察其对多种肿瘤细胞增殖的抑制作用。方法以粒径、Zeta电位、多分散性指数(PDI)为指标,考察稳定剂、药载比、超声温度、均质温度、均质次数对西瑞香素纳米混悬剂反溶剂沉淀法制备的影响,优化最佳处方和工艺条件。观察西瑞香素纳米混悬剂的透射电镜表征,并进行X射线衍射、差示扫描量热分析,考察其在血浆、PBS中的稳定性以及载药量、体外释放情况,采用MTT法比较其对BT474、SKBR-3、A549、He La、Hep G2细胞的体外细胞毒性。结果最佳处方和工艺条件:TPGS为稳定剂,药载比1∶1,共同溶解于DMSO中,在超声(25℃,250 W)条件下,缓慢滴注于去离子水中,透析除去有机溶剂,高压均质(25℃,150 MPa)20次,即得西瑞香素纳米混悬剂,其平均粒径为(163.1±5.4)nm,Zeta电位为(-11.4±0.7)m V,PDI为0.15±0.04。西瑞香素纳米混悬剂近乎为球形,大小较均匀;在血浆中稳定存在,不存在溶血现象,满足静脉注射需求,平均载药量为(39.16±1.09)%。西瑞香素纳米混悬剂对5种受试细胞的生长抑制作用显著提高,并呈现剂量相关性,尤其是对SKBR-3、A549、He La、Hep G2细胞,IC_(50)在2.1~3.4μg/m L。结论西瑞香素纳米混悬剂具有小粒径、高载药量、显著肿瘤细胞毒性等优点,在抗肿瘤研究方面具有良好的应用前景。     

西瑞香素对肺癌A549细胞侵袭及迁移能力的影响

目的探讨西瑞香素对肺癌A549细胞侵袭及迁移能力的影响。方法通过Transwell小室分析西瑞香素对肺癌A549细胞侵袭能力的影响;划痕实验检测西瑞香素对肺癌A549细胞迁移能力的影响;Western blotting检测西瑞香素作用于A549细胞后MMP-2、MMP-9侵袭相关蛋白的表达。结果 Transwell小室实验结果提示,西瑞香素对A549细胞侵袭能力有抑制作用;划痕实验结果提示,西瑞香素对A549细胞迁移能力有抑制作用;Western blotting结果提示,西瑞香素抑制MMP-2、MMP-9侵袭相关蛋白的表达。随着药物浓度增加,MMP-2、MMP-9表达减少,具有剂量依赖性。结论西瑞香素对A549细胞体外侵袭及迁移有抑制作用,这种作用与抑制MMP-2、MMP-9表达有关。     

高效液相色谱-四级杆飞行时间串联质谱法分析瑞香素在大鼠肠壁的代谢产物

研究瑞香素在大鼠肠壁产生的代谢产物。采用大鼠在体肠灌流模型,分别收集瑞香素0～2 h内的十二指肠、空肠、回肠和结肠灌流液,以液相色谱-四级杆飞行时间串联质谱法分析肠道灌流液中瑞香素的代谢产物。在大鼠十二指肠、空肠、回肠灌流液中共发现瑞香素原形药物和4个代谢产物,分别推测为瑞香素-7-硫酸酯、瑞香素-8-硫酸酯、瑞香素-7-葡糖醛酸结合物和瑞香素-8-葡糖醛酸结合物;而在结肠灌流液中未发现代谢产物。在瑞香素的4个代谢产物中,瑞香素-7-硫酸酯和瑞香素-8-硫酸酯(m/z 257)为首次发现的瑞香素在大鼠体内的Ⅱ相代谢产物,揭示了其在大鼠体内代谢的新途径。     

西瑞香素通过NF-κB途径抑制RANKL诱导的破骨细胞分化

摘要：目的:研究西瑞香素作为一种具有抗炎抗氧化作用的中药成分对破骨细胞分化的影响及机制。方法:从BALB/c小鼠骨髓中分离获得骨髓单核细胞,使用GST-rRANKL诱导形成破骨细胞并分为3组:对照组、RANKL诱导组和RANKL+西瑞香素组,采用TRAcP染色评估西瑞香素对破骨细胞成熟与融合的影响,用Western Blot和qRT-PCR检测破骨细胞相关表型蛋白和核因子κB（NF-κB）信号通路蛋白,构建荧光素酶报告基因检测转录因子NF-κB活性。结果:西瑞香素能够显著抑制RANKL诱导的破骨细胞形成,抑制RANKL刺激后的Acp5、V-ATPase-d2和CTSK的转录水平,并抑制与破骨细胞分化密切相关的两个转录因子NFATc-1和c-Fos的表达量。西瑞香素能促进IκB-α的表达,从而抑制NF-κB活性。结论:西瑞香素通过促进IκB-α的表达,抑制NF-κB活性,使破骨细胞分化相关的关键转录因子NFATc-1和c-Fos的表达量下调,从而通过抑制NF-κB途径发挥抑制RANKL诱导的破骨细胞分化的作用。     

西瑞香素与大鼠血浆蛋白结合率的测定

目的采用HPLC法对西瑞香素与大鼠血浆蛋白结合率进行研究。方法采用平衡透析法,结合HPLC法测定西瑞香素的大鼠血浆蛋白结合率。结果西瑞香素在低(0.50 mg.L-1)、中(1.0 mg.L-1)、高(2.0 mg.L-1)3个质量浓度下,其血浆蛋白结合率分别为(91.7±1.8)%、(91.6±1.4)%和(90.7±0.81)%。结论西瑞香素具有较强的血浆蛋白结合率。     

RP-HPLC-UV法测定大鼠血浆中西瑞香素浓度(英文)

建立了测定大鼠血浆中西瑞香素浓度的高效液相色谱法,并对其进行方法学确证。以蛋白沉淀法和液液萃取法对大鼠血浆中西瑞香素进行提取。采用Diamonsil C_(18)柱(200mm×4.6mm,5μm),流动相为甲醇-20mmol/L醋酸铵(醋酸调pH至3.2,42:58,v/v),流速1.0mL/min。检测波长345nm,柱温40℃。西瑞香素在0.020-2.00μg/mL浓度范围内线性关系良好(r>0.9900),定量下限0.020μg/mL。日内和日间精密度(RSD)为5.0%-10.6%,准确度(RE)为±(1.2%-2.5%)。所建立的方法成功应用于腹腔注射西瑞香素在大鼠体内的药代动力学研究。     

瑞香素缓释片中瑞香素含量测定方法的建立与验证

目的建立一种测定瑞香素缓释片中瑞香素含量的高效液相色谱法（HPLC）并对其进行验证。方法色谱柱为Agela Venusil C18（4.6 mm×250 mm,5μm）,流动相为甲醇-0.05%磷酸溶液（45∶55）,流速1.0 ml/min,柱温30℃,检测波长323 nm,进样量10μl。结果瑞香素在5～45μg/ml范围内具有良好的线性关系,R2=0.9999,平均回收率为101.17%,RSD为0.3%。结论该法准确可靠,操作简便,可用于瑞香素缓释片的含量测定。     

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.