牛和羊胎盘肽的制备及其生物活性的研究

目的探索一种新型的免疫调节剂,充分利用动物胎盘。方法利用超滤法从牛和羊胎盘中提取胎盘肽,采用改良的加脲Tricine-SDS-PAGE法测定其相对分子质量;利用紫外光扫描其最大吸收峰;并通过建立体外抑制兔淋巴细胞模型,利用四甲基偶氮唑盐(MTT)比色法测定所提取胎盘肽的免疫调节作用。结果从牛和羊胎盘中成功提取出了胎盘肽,其相对分子质量分别为4 785和4 386;其最大吸收峰分别为210.5和225.4 nm;牛和羊胎盘肽均可使顺铂抑制的兔外周血T淋巴细胞的转化率显著提高,其中牛胎盘肽以1∶1×103、1∶1×1042组吸收度明显高于抑制组(P<0.05);羊胎盘肽以1∶1×102、1∶1×103和1∶1×1043组效果最为明显(P<0.01)。结论牛和羊胎盘中含有的胎盘肽可成功提取出来,其可明显提高淋巴细胞的转化率,是一种较为理想的免疫调节剂。     

致倦库蚊不同发育期体内可溶性蛋白的分析研究

应用SDS—聚丙烯酰胺凝胶电泳技术,分析测定致倦库蚊不同发育体内的可溶性蛋白,结果显示致倦库蚊成蚊显27条蛋白区带,其主要蛋白带的分子量分別为81KDa、64KDa、50KDa;蛹:显现28条蛋白区带,主要蛋白带的分子量分別为93KDa、45KDa和10KDa;Ⅳ龄幼虫显现31条蛋白区带,主要蛋白带的分子量分别是85KDa、76KDa和47KDa.证实这些区带即为各自特异性蛋白区带.     

羊胚胎盘肽功能测试

目的探讨羊胚胎盘肽的制备及生物活性。方法应用膜技术制备羊胚胎盘肽 ,对羊胚胎盘肽的含量、肽谱、氨基酸、活性、紫外吸收等进行测试 ,利用动物实验观察羊胚胎盘肽对调节免疫、延缓衰老的作用。结果羊胚胎盘肽是一组小分子多肽 ,分子量小于 10 0 0 0D的占 37.3% ,紫外吸收峰在 194nm。结论羊胚胎盘肽具有调节免疫和延缓衰老的生物学活性     

羊红细胞铜锌超氧化物歧化酶的纯化及部分性质研究

目的从羊红细胞中分离纯化铜锌超氧化物歧化酶 (Cu ,Zn SOD) ,并对其部分理化性质进行研究。方法采用有机溶剂去除血红蛋白、热变性、超滤浓缩、丙酮沉淀、DE FF柱色谱的方法 ,对羊红细胞Cu ,Zn SOD进行分离纯化。结果羊红细胞 5 2 0g得Cu ,Zn SOD总活力为 4 6 70 0 0u ,比活为 812 6u/mg·pro,纯化倍数为 2 6 .2 ,活性回收率为 6 1%。理化性质分析表明 :该酶的最大紫外吸收波长为 2 5 8nm ,亚基相对分子量为 16 .1kD ,每个亚基含 1个铜原子和 1个锌原子。pH在 6～ 11范围内该酶稳定性很好 ,在 35℃～ 75℃范围内 ,保温 2 0min ,酶活基本没有损失。 2mmol/L的SDS对Cu ,Zn SOD没有明显的抑制作用 ,而 2mmol/L的H2 O2 对该酶表现出较强的抑制作用。结论采用此方法分离纯化的Cu ,Zn SOD达到电泳纯 ,其理化性质与其它动物血液来源的Cu ,Zn SOD一致。     

纤维素酶辅助提取黄花蒿中青蒿素的工艺条件研究

目的 研究纤维素酶辅助提取青蒿素反应中各条件对提取率的影响.方法 采用紫外分光光度法测定青蒿素含量,研究酶反应过程中时间、加酶量、温度和pH值对提取率的影响,并使用正交实验方法对提取工艺进行优化.结果 纤维素酶辅助提取青蒿素的最优工艺条件分别是:酶反应时间2h,纤维素酶用量0.30 g,酶反应温度45℃,酶反应pH值4.5.结论 纤维素酶辅助提取青蒿素的方法能够有效提高提取率.     

蛹虫草子实体中多肽的提取分离及组成初探

目的获取蛹虫草子实体中的肽并对其分离纯化。方法采用水提醇沉法提取,葡聚糖凝胶色谱法分离纯化,继而用氨基酸自动分析仪分析组成。结果由单因素考察和正交试验法确定了最佳提取条件,经分离纯化后得到两个小分子量的肽组分,分别测定出了两组分的氨基酸组成及含量。结论分离出的肽类组分氨基酸种类丰富齐全,具有一定的营养和药用价值。     

正交设计法优选骨肽注射液纯化参数的研究

目的:探讨优选骨肽注射液提取的工艺条件。方法:采用正交设计法,骨肽注射液多肽含量及操作可行性为考察指标,对骨肽注射液提取过程中乙醇沉淀浓度、酸沉淀(pH 2.0～4.0)、碱沉淀(pH 6.8～7.2)3个因素进行筛选。结果:对于选定的三因素三水平,骨肽注射液最佳纯化参数组合为乙醇沉淀浓度70%、酸沉淀pH 4.0,碱沉淀pH 8.0,结果具有较显著的统计学意义。结论:根据骨肽注射液最佳提取条件对猪骨进行骨多肽提取,提取效率高,成本低。     

羊胎盘肽对小鼠非特异性免疫功能的影响

目的探讨羊胎盘肽对小鼠非特异性免疫功能的影响。方法采用60Coγ射线照射的方法建立免疫抑制小鼠动物模型,利用巨噬细胞吞噬鸡红细胞和小鼠碳粒廓清实验,评价羊胎盘肽对免疫抑制状态小鼠非特异性免疫功能的影响。结果羊胎盘肽有促进巨噬细胞吞噬鸡红细胞功能的作用(P<0.05),并能显著提高机体碳粒廓清的能力(P<0.01)。结论羊胎盘肽对小鼠具有一定非特异性免疫增强作用。     

尿激酶溶液及其冷冻干燥制剂的稳定性研究

<正> 尿激酶在临床上主要用于溶血栓。本文对应用组织培养技术,从人肾细胞中分离得到的低分子量尿激酶(分子量为33,000)在溶液中及在冷冻干燥后的稳定性进行了研究。研究表明,尿激酶在55℃,pH5～9的溶液中的失活符合伪一级动力学过程。在pH6.5时最稳定。以前曾有报道,从尿中提取的低分子量尿激酶在pH9左右时最稳定,而高分子量尿激酶在pH3～9时均相当稳定。因此,作者     

淫羊藿苷及其制剂含量测定研究概况

淫羊藿为常用中药,系小檗科植物淫羊藿的干燥地上部分。有补肾壮阳、祛风除湿的作用。淫羊藿苷是淫羊藿的主要成份,其含量测定对于研究药物的药理,临床疗效,控制制剂质量等均有重要意义。笔者查阅了近几年国内的含量测定方法,综述如下。1.紫外分光光度法(UV)紫外分光光度法测定淫羊藿苷基本采用有机溶剂提取分离再行测定。为了考查淫羊藿甙的提取工艺王昌利等[1]用紫外分光光度测定了淫羊藿提取液乙酸乙酯甙的含量,水提液浓缩,用乙酸乙酯萃取4次,回收乙酸乙酯,用甲醇溶解在267nm测定。骨质增止痛液由淫羊藿等7味中药组成,赵春香等[2]应用大…     

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.