鱿鱼皮胶原蛋白医用止血材料的研究

目的 从鱿鱼皮中提取胶原蛋白并研究经化学和物理方法改性的鱿鱼皮胶原海绵的止血效果。方法 从新鲜鱿鱼皮中提取得到酸溶性胶原蛋白 (ASC)和酶溶性胶原蛋白 (PSC),冷冻干燥制备胶原海绵,分别采用EDC交联、干热交联 (DHT)、EDC/DHT结合交联这3种方法对其进行改性处理。采用兔耳创伤止血和肝脏止血试验来评价改性后胶原海绵的止血性能。结果 鱿鱼皮胶原蛋白ASC和PSC为典型的I型胶原蛋白。动物止血实验结果表明ASC-E、PSC-E、ASC-E/D和PSC-E/D等4种改性鱿鱼皮胶原海绵组均能对创面起到一定止血作用,其中尤以PSC-E组的止血效果最好,优于市售明胶海绵。 结论ASC-E、PSC-E、ASC-E/D和PSC-E/D等4种改性鱿鱼皮胶原海绵组均能有效的缩短出血时间,减少出血量,达到快速止血的效果,其中PSC-E止血效果最佳。     

大马哈鱼皮胶原蛋白制备和表征

用稀碱处理大马哈鱼皮,然后进行限制性酶解,制备得到胶原蛋白,检验制备样品的纯度,测定样品的热变性温度和细胞相容性。制备的胶原蛋白α链的含量≥75%,β链的含量≤25%,有微量的γ链;变性温度是12.5℃;细胞培养3天的相对活力是80.0±3.6%;大马哈鱼皮胶原蛋白为基质的细胞活力仅次于对照组(空白细胞培养板),优于聚乳酸、聚羟基丁酸和聚-3-羟基丁酸戊酸。海鱼皮胶原蛋白可进一步应用于生物材料研究。     

不同鲨鱼皮胶原蛋白的分离及其特性研究

目的 探讨不同鲨鱼皮胶原蛋白的分离方法,并对分离胶原蛋白的部分生化特性进行研究.方法 用稀碱溶胀酶解法争缓冲液溶胀分离法分别从水鲨头皮和马鲛鲨皮中分离胶原蛋白,利用聚丙烯酰胺凝胶电泳法(SDS-PAGE)、差示扫描量热仪法(DSC)、园二色谱仪法(CD)对分离胶原蛋白进行纯度及分子大小、变性温度和二级结构的测定.结果 鲨鱼皮胶原蛋白回收率为鲜重的2.9%～4.1%;分离胶原蛋白在SDS-PAGE上呈现清晰的3条谱带,相对分子质量分别为205,134,118 KDa水鲨头皮和马鲛鲨皮胶原蛋白的热变性温度分别为69℃、47℃;两种分离胶原蛋白的二级结构均主要是β-折叠和无规卷曲,不存在α-螺旋.结论 采用稀碱溶胀酶解法和缓冲液溶胀分离法均能制备高纯度的胶原蛋白,从水鲨头皮和马鲛鲨皮分离的胶原蛋白相对分子质量争分子构成一致,但提取条件的不同能影响胶原蛋白的热变性温度和二级结构.     

毛尖蘑粗多糖提取方法的研究

目的：优化毛尖蘑子实体粗多糖提取方法,探讨毛尖蘑粗多糖提取的最佳工艺。方法：采用热水浸提法对毛尖蘑中的粗多糖进行提取,比较在不同浸泡温度下毛尖蘑粗多糖的提取率。结果：温度在80℃、90℃、100℃时,提取物经过漂洗,毛尖蘑粗多糖提取率分别为3.712%、2.794%、2.040%;温度在80℃、90℃时,提取物未经过漂洗,毛尖蘑粗多糖提取率分别为1.530%、0.965%。结论：提取毛尖蘑子实体粗多糖的最佳温度是80℃,同时,提取物经漂洗后提取率提高。     

酵母发酵鲜党参中多糖的提取及免疫活性研究

目的 优化酵母发酵鲜党参中多糖的提取工艺,并评价其体外免疫活性。方法 以酵母发酵鲜党参为原料,通过木瓜蛋白酶辅助提取,乙醇沉淀得到酵母发酵鲜党参中多糖,以提取温度、时间、酶添加量为影响因素,多糖得率为指标,以水提法为对照,进行单因素实验结合响应面设计开展工艺优化,通过高效凝胶渗透色谱法测定多糖的相对分子量,通过MTT法研究多糖对体外脾淋巴细胞和RAW264.7细胞的增殖活性。结果 酵母发酵鲜党参中多糖的最佳提取工艺为提取温度63 ℃,提取时间6 h,酶添加量4%,多糖的提取率17.46%,比水提法(12.69%)提高。且糖醛酸含量提高。酶提法和水提法得到的多糖相对分子量无明显变化。当多糖浓度为0.5,1.0 mg·mL^-1时,给予酶提多糖的脾淋巴细胞增殖活力和RAW264.7细胞增殖活力均比水提法增加。结论 本研究优选出的酵母发酵鲜党参中多糖的提取工艺切实可行,多糖提取率高且免疫活性显著增强。     

酶法提取杜仲中降压活性成分

目的采用酶法提取杜仲中降压活性成分。方法选用纤维素酶,通过L9(34)正交试验确定京尼平苷酸和京尼平苷最佳提取工艺条件。结果提取京尼平苷酸的最佳工艺参数:温度45℃,介质pH=5,酶解时间1h,每100g杜仲需纤维素酶0.3g,提取率达到1.58%;提取京尼平苷的最佳工艺参数为:温度45℃,介质pH=7,酶解时间0.5h,每100g杜仲需纤维素酶0.2g,提取率达到0.26%。结论与传统水浸提取工艺相比,提取率提高9.09%~14.45%。     

响应面法优化生姜多糖的提取工艺研究

目的：采用响应面法优化法优化生姜多糖的提取工艺〔1〕。方法对醇沉水提法提取生姜多糖的工艺条件是运用 Box-Behnken中心组合响应面分析法优化,主要探讨影响生姜多糖提取率的3个主要因素即生姜多糖提取过程的提取温度、提取时间及提取料液比对生姜多糖提取率的作用。结果在此次实验条件下,采用醇沉水提法提取生姜多糖的最佳工艺条件应该是：提取时间118min,提取温度89℃,提取料液比为1∶15,生姜多糖的提取率可以达到4.62％。结论此工艺方便简单可行,是一条提取生姜多糖的的有效方法。     

酶法提取苦豆草生物碱研究

目的在实验室酶法提取的基础上,优化苦豆草生物碱酶法提取小试工艺条件。方法以苦豆草生物碱的提取率为指标,在小试条件下优化提取次数、料液比、提取时间,并与传统提取法比较。结果酶法提取最佳的小试工艺条件为:酶解时间4h,溶剂pH=6,温度为50℃,加酶量为1∶1 000,酶解后,加入盐酸至浓度为4mL·L-1,料液比1∶16,提取2次,第1次2h,第2次1h。苦豆草生物碱的提取率达到2.17%结论实验室提取工艺与小试生产存在差异,经条件优化基本达到实验室提取生物碱的提取率。     

Ⅱ型胶原蛋白的热稳定性、圆二色性和红外光谱研究

目的以蓝鲨Ⅱ型胶原蛋白为材料,分析Ⅱ型胶原蛋白的热稳定性、圆二色性和红外光谱特性。方法采用限制性酶解法分离蓝鲨软骨Ⅱ型胶原蛋白,通过SDS-PAGE、DSC、CD、IR分析Ⅱ型胶原蛋白的相对分子质量、热变性温度、二级结构特征和红外光谱特征。结果蓝鲨Ⅱ型胶原蛋白是由相对分子质量为130kDa的α1链构成;蓝鲨Ⅱ型胶原蛋白的热变性温度为40.5℃,[Na+]和[H+]改变了胶原蛋白的热稳定性;蓝鲨Ⅱ型胶原蛋白在3411、3008、1648、1549cm-1有红外特征性吸收峰;蓝鲨Ⅱ型胶原蛋白以β-折叠为主,占主链的36.9%,接近于全β型三级结构,[H+]将改变Ⅱ型胶原蛋白的二级结构元件构成,使其β-折叠消失,出现少量的α-螺旋,Ⅱ型胶原蛋白肽链转变成无规卷曲。结论蓝鲨Ⅱ型胶原蛋白是典型的纤维状蛋白质,具有作为生物功能物质的优良生物化学特性。     

响应面法优化酶法提取款冬花多糖工艺研究

目的 优化款冬花多糖酶法辅助提取工艺条件。方法 通过响应面法,以多糖提取率为评价指标,考察提取时间、酶药质量比和温度等因素对款冬花多糖提取的影响。结果 最优工艺条件为：55℃温度下,用20倍量的水,选择酶药质量比1.30%,提取123.2min,pH为7,水料比为20(mL/mg)。在此工艺条件下,款冬花多糖的提取率为3.25%。结论 酶法提取款冬花多糖具有提取温度低,提取率高的优点。     

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.

     

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.     

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.     

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.