从猪胰制备甘油三酯测定用脂肪酶

从猪胰制取测定甘油三酯(TG)的脂肪酶,其活性大于2.7×10~4u/mg;比活为5.5×10~4u/mg 蛋白;酶作用的最佳 pH、最适反应时间及最适温度分别为7.5～10.0、15min 及37～50℃。聚丙烯酰胺梯度凝胶电泳测得分子量为34700及20900;以橄榄油为底物的米氏常数为0.77%。     

用提纯干扰素预防鼻病毒感染

本文报道一项用提纯干扰素(1.6×10~8U/mg)预防鼻病毒感染的双盲对照研究。用单克隆抗体亲和层析提纯的人白细胞干扰素给志愿者作鼻腔喷雾,每个鼻孔喷干扰素2×10~6U,共7次,每次间隔4～16小时,喷后5～60分钟取鼻洗液,测干扰素活性。喷2×10~6U 干扰素时,5分钟后 RIA 分     

从培养的香菇菌丝中提取抗病毒及诱导干扰素的新多肽甘露聚糖:KS-2

具有抗肿瘤活性的免疫增效剂KS-2是由香菇菌丝培养物中提取而得,含有α-甘露糖与小肽相连,分子量为6×10~4～9.5×10~4,小鼠口服急性毒性LD_(50)大于12,500mg/kg。小鼠或大鼠口服或腹腔注射KS-2,通过宿主机体免疫功能的提高可抑制各种转移瘤的生长。DDI小鼠口服或腹腔注射KS-2200mg/kg,16小     

氟伐他汀致肝损害1例报告

肝功能损害的定义为丙氨酸氨基转移(ALT)和天冬氨酸基转移(AST)2项指标超出正常值3倍。患者男,52岁,体形偏胖,既往无特殊病史。于去年8月在我院健康体检中血生化指标显示,总胆固醇(TC)17.8mmol/L(正常3.9~5.7mmol/L),三酰甘油(TG)11mmol/L(正常0.6~2.2mmol/L)。肝酶学检查,天门冬氨酸基转移酶(AST)30U/L(正常8~40U/L),丙氨酸氨基转移酶(ALT)35U/L(正常8~42U/L)。肝炎血清病毒学检查结果全部阴性(包括甲肝、乙肝、丙肝)。考虑到患者TC及TG均高,遂给予氟伐他汀40mg、连服10d后复查血生化:TC6.5mmol/L、TG3.1mmol/L、AST188U/…     

对硫磷抑制Ca~(2 )—ATPase的动力学分析

作者研究了对硫磷对人红细胞的Ca~(2 )—ATPase的抑制动力学。结果表明,在无对硫磷存在的情况下,正常对照 Ca~(2 )—ATPase的Km值为1.3×10~(-4)mol/L,Vmax为28.9nmol pi/mg蛋白/min,而当反应体系中有 2×10~(-4)～2×10(-3)mol/L对硫磷存在时,则Km增加为1.5～2.4×10~(-4)mol/L,Vmax却减小为27.3～18.46nmol pi/mg蛋白/min。用Hanes法作图结果提示,对硫磷对人红细胞Ca~(2 )—ATPase 的抑制是一种混合性的抑制作用。     

人尿胰蛋白酶抑制剂的快速纯化

目的以人尿浓缩物为原料 ,对人尿胰蛋白酶抑制剂的分离纯化技术进行研究。方法利用纯胰蛋白酶作为配体偶联于CNBr Sepharose 4B为亲和载体 ,在不同条件下洗脱亲和蛋白 ,可从粗品溶液中快速分离纯化具有胰蛋白酶抑制活性的两种纯产物。结果层析图谱与SDS PAGE分析表明 ,根据洗脱条件 ,可将有抑制活性的 2 0kD和 40kD蛋白质分离为两个单独组份或混合物。其中 2 0kD组份的抑制活性回收为 35 % ,抑制比活为 3 387U/mg;40kD组份的相应值分别为 5 3 %和 3 116U/mg。混合物的抑制活性回收和抑制比活分别为 89%和 3 0 76U/mg ,再经Su perose 12或Superdex 75凝胶渗透层析 ,分别获得两种组份的纯产物。 结论通过选择性洗脱亲和柱 ,可快速获得高收率、高纯度的人尿胰蛋白酶抑制剂     

新法提取脲酶的研究

介绍了从大豆中提取脲酶的新方法，摒弃了传统的柱色谱分离、浓缩、透析等繁琐步骤。制得的脲酶活性大于５０Ｕ／ｍｇ，比活为６１．２Ｕ／ｍｇ蛋白；酶作用最佳ｐＨ为６．７，最适反应温度为３７℃；以尿素为底物的米氏常数Ｋｍ为３．６７×１０－３ｍｏｌ／Ｌ，固体在冰箱中贮存一年活性基本不变     

肝素诱导血小板减少症2例

例1为24岁男性患者,因急性心肌梗死,给予阿司匹林300mg、氯吡格雷300mg口服和普通肝素10000U静脉推注,并行冠状动脉造影、经皮冠状动脉腔内成形术及支架植入术。患者术前血小板计数为228.0×10⁹/L,术后1h降至36.2×10⁹/L。考虑为肝素诱导的血小板减少,遂停用肝素,给予阿加曲班0.5～2μg·kg^-1·min^-1静脉滴注。第3天患者血小板升至101×10⁹/L,第4天恢复正常。例2为69岁男性冠状动脉粥样硬化性心脏病患者,在冠状动脉造影术中静脉推注普通肝素3000U。之后,行冠状动脉搭桥术,术前连续7d给予低分子肝素(1mg·kg^-1·12h^-1)皮下注射,术中给予普通肝素7000U静脉滴注。术前患者血小板计数197.0×10⁹/L,术后降至19.2×10⁹/L。停用肝素,给予阿加曲班0.5～1.5μg·kg^-1·min^-1静脉滴注,血小板计数升至146.0×10⁹/L。     

酰胺菌素与头孢西丁,庆大霉素,甲硝唑或妥布霉素静脉混合注射液的稳定性

取酰胺菌素(Aztreonam,A)的粉针,庆大霉素(G)注射液(40mg/ml,10mg/ml),妥布霉素(T)注射繁(40mg/ml,10mg/ml),头孢西丁(Cefoxitin,C)注射液(95mg/ml),以0.9%氯化钠或5%葡萄糖注射液为稀释剂,分别配制成4种不同浓度的混合液。另取A粉针及甲硝唑(灭滴灵)(M)注射液(5mg/ml)以磷酸-枸橼酸缓冲液配制成2种浓度的混合液,所有样品均置4℃(冰箱)7天或25℃(室温)48h,于不同贮存时间,取     

磺苄西林对粘液型和非粘液型绿脓杆菌的体外抗菌活性

用试管双倍稀释法测定磺苄西林(Sulbenicil-lin)、羧苄青霉素、氨苄青霉素、哌拉西林(Piperacil-lin)和头孢噻肟对20株粘液型和50株非粘液型绿脓杆菌的体外抗菌活性。结果表明,细菌接种量为10~(-4)CF U/ml 时,磺苄西林的抗菌活性较羧苄青霉素强,但较哌拉西林和头孢噻肟弱。显然,这些细菌对受试的β-内酰胺抗生素具高度耐药性,尤当细菌接种量为10~7CF U/ml 时更为明显。哌拉西林和头孢噻肟在接种量为10~7 CFμ/ml 时的抗菌活性较10~4时为小,这在杀菌浓度时更为明显。磺苄西林、羧苄青霉素和头孢噻肟的MBC/MIC 比率范围为1～2,而哌拉西林则为3～     

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.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

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.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.