定量检测细菌内毒素的鲎试验

目的：建立一种简便,可靠的内毒素定量检测方法。方法：以组氨酸包被酶标反应板,利用固化的组氨酸吸附分离样品中的内毒素,最后以显色鲎试剂定量检测被吸附的内毒素。结果：预包被的组氨酸能有效地吸附样品中的内毒素,但不减弱内毒素的鲎试剂反应性,本法能较好地克服β-1.3-D-glucan、氨基酸、抗生素及血浆蛋白等对鲎试验（LAL）的干扰,并具有较高的准确性和重现性。结论：本法可用于各种含LAL干扰物的样品中内毒素检测。     

应用LAL动态浊度法测定复方苦参注射液中细菌内毒素含量

目的：建立应用美洲鲎试剂LAL的动态浊度法定量检测复方苦参注射液中细菌内毒素试验方法。方法：采用《中国药典》2015年版(四部)中的细菌内毒素检查法。结果：复方苦参注射液用细菌内毒素定量法检测无干扰因素影响,内毒素回收率在50%～200%范围内。结论：使用LAL进行动态浊度法定量检测复方苦参注射液的细菌内毒素是可行的,可用细菌内毒素方法代替兔热原检查法,同时又避免了限量法内毒素检测时的干扰。     

固化组氨酸吸附鲎试验定量检测内毒素

建立一种简便、可靠的内毒素定量检测方法。方法：在含干扰物的样品中,利用固化组氨酸吸附样品中的内毒素,再以基质偶氮显色法鲎试验定量检测被吸附的内毒素。结果：该法能较好地克服氨基酸、抗生素等可致鲎试验假阴性物质的干扰,并具有较高的准确性和精密度。结论：此法是一种简便、特异的内毒素定量检测方法。     

注射用盐酸赖氨酸细菌内毒素检查方法学研究

目的:研究注射用盐酸赖氨酸对细菌内毒素检查试验的干扰情况,建立其细菌内毒素限量检查的方法.方法:采用细菌内毒素凝胶法,用两个厂家的鲎试剂对提供的注射用盐酸赖氨酸连续3批样品进行干扰试验.结果:注射用盐酸赖氨酸最大非干扰浓度为100 mg/mL.结论:注射用盐酸赖氨酸用细菌内毒素检查法进行细菌内毒素检测是可行的,其内毒素限值为每1 mg盐酸赖氨酸含内毒素量应小于0.01 EU.     

动态浊度法检测关节腔内注射用交联透明质酸钠中细菌内毒素

目的:建立定量检测关节腔内注射用2%交联透明质酸钠中细菌内毒素的试验方法。方法:根据2010年版《中国药典》附录细菌内毒素检查法中的动态比浊法及其指导原则进行干扰试验,确定最大有效稀释倍数;并将检测结果与凝胶法比较。结果:样品在原液8倍稀释的浓度下用定量鲎试剂检测无干扰,内毒素回收率在50%²00%内;两种方法检测样品内毒素结果均在规定限值下。结论:建立的动态比浊法定量检测关节腔内注射用2%交联透明质酸钠中的内毒素方法可行。     

3种中药注射剂细菌内毒素检查方法研究

目的：建立3个品种中药注射剂的细菌内毒素检查方法,比较基因重组鲎试剂（rLAL）与传统鲎试剂（TAL/LAL）检查结果的差异,确定rLAL的适用性。方法：依据《中国药典》2020年版四部<1143 细菌内毒素检查法>、<9251细菌内毒素检查法应用指导原则>,采用rLAL、LAL、TAL3种鲎试剂对丹红注射液、痰热清注射液、血必净注射液3种中药注射剂进行细菌内毒素检查。结果：3种鲎试剂均可用于3种中药注射剂细菌内毒素检查,检测结果均符合规定。其中rLAL检测灵敏度更高,可达0.001 EU·mL-1,且具有更大的MVD值。结论：基因重组鲎试剂可取代传统鲎试剂用于3种中药注射剂细菌内毒素检查。     

动态浊度法定量检测醒脑静注射液中的细菌内毒素

目的：建立动态浊度法测定醒脑静注射液中细菌内毒素含量的方法。方法：依据《中国药典》2020年版四部通则（1143细菌内毒素检查法）中的动态浊度法及其指导原则,分别应用美洲鲎试剂和安度斯鲎试剂,在Pyros Kinetix? Flex（PKF）细菌内毒素定量检测系统上,对醒脑静注射液中的细菌内毒素进行方法开发。通过细菌内毒素标准曲线可靠性试验和干扰预实验确定有效稀释倍数,测定干扰试验的回收率,完成醒脑静注射液细菌内毒素的定量检测。结果：可靠性试验中,LAL标准曲线浓度线性范围为0.001～1.0 EU·mL－1（r=-0.987 7）,标准曲线的回归方程为：lgT=-0.312 3lgC+2.740。TAL标准曲线浓度线性范围为0.01～1.0 EU·mL－1（r=-0.999）,标准曲线的回归方程为：lgT=-0.270lgC+2.830。供试品在稀释比例1：20及以下浓度时对鲎试剂反应无干扰作用;3批供试品的内毒素定量检测结果均低于供试品规定限值。结论：应用Pyros Kinetix? Flex（PKF）细菌内毒素定量检测系统,动态浊度法可用于醒脑静注射液细菌内毒素含量的定量检测。     

含葡萄糖甘露醇注射液细菌内毒素检查方法的研究

本文对用细菌内毒素检查法检测含葡萄糖甘露醇注射液中细菌内毒素的可行性进行了研究。结果表明,含葡萄糖甘露醇注射液稀释４倍后对鲎试剂没有干扰作用,可选用灵敏度标示值为０．１２５Ｅｕ／ｍｌ的鲎试剂检测样品的细菌内毒素。     

含葡萄糖甘露醇注射液细菌内毒素检查方法的研究

本文对用细菌内毒素检查法检测含葡萄糖甘露醇注射液中细菌内毒素的可行性进行了研究。结果表明 ,含葡萄糖甘露醇注射液稀释 4倍后对鲎试剂没有干扰作用 ,可选用灵敏度标示值为 0 .12 5Eu/ ml的鲎试剂检测样品的细菌内毒素     

注射用盐酸头孢吡肟细菌内毒素检查方法探讨

目的：探讨注射用盐酸头孢吡肟细菌内毒素检查方法的可行性。方法：通过干扰初筛试验及干扰试验确定可选用灵敏度高于0．06EU／mL鲎试剂检测注射用盐酸头孢吡肟的细菌内毒素；再通过鲎试剂灵敏度的复核、样品热原法及细菌内毒素检查法进一步确定该药细菌内毒素检查方法的可行性。结果：本品采用λ=0．06EU／mL的鲎试剂，在1.0mg／mL稀释浓度下无干扰作用，L=0.06EU／mg。结论：选用灵敏度=0．06EU／mL的鲎试剂能准确检测注射用盐酸头孢吡肟细菌内毒素。     

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.     

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.     

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.