微量生色底物法测定肝素钠抗Ⅹa因子和抗Ⅱa因子活性

目的英国药典中收载了半微量生色底物法测定低分子肝素抗Ⅹa因子和抗Ⅱa因子活性,在此基础上,我们考虑是否可以将实验方法进行改进,用微量生色底物法测定肝素类药物活性。方法调整样品与试剂浓度,降低总反应体积,在96孔酶标板中进行反应,用酶标仪测定样品吸光度。结果对6个肝素样品进行抗Ⅹa因子和抗Ⅱa因子活性测定,实验结果误差小,重复性良好。结论微量生色底物法可以用于肝素类药物抗Ⅹa因子和抗Ⅱa因子活性测定,该法提高了实验效率,降低了检验成本,可以考虑用本法取代传统的凝血法测定肝素效价。     

D-聚甘酯的分子量与抗FⅩa/抗FⅡa活性比值的相关性研究

目的 观察不同分子量的D-聚甘酯(DPS)对大鼠抗FⅩa/抗FⅡa活性比值的影响.方法 将30只大鼠随机分为5组,分别给予生理盐水、低分子肝素(360 U·kg-1)及3种不同分子量的DPS(9 mg·kg-1).给药后1h,分别测定血浆的APTT、PT、FⅠB、ATⅢ及抗FⅩa、抗FⅡa活性.结果 高剂量DPS组的抗FⅡa活性高于低分子肝素及中、低剂量DPS组,抗FⅩa/抗FⅡa比值明显低于其余各组.结论不同分子量的D-聚甘酯,抗FⅩa活性无显著性差异.分子量增加,抗FⅡa活性增强,抗凝作用增强;分子量减小,抗FⅩa/抗FⅡa比值增大,抗栓作用增强.     

低分子肝素相对分子质量与抗凝活性关系研究*

目的:研究低分子肝素相对分子质量与抗凝活性之间的关系。方法:用生色底物法测定不同相对分子质量的低分子肝素6个分级和未分级样品的抗Ⅹa因子和抗Ⅱa因子效价,相对分子质量采用高效体积排阻色谱法(high performance size exclusion chromatography,HPSEC)测定。结果:抗Ⅹa因子与抗Ⅱa因子效价比随着相对分子质量的降低而增大。结论:在测定范围内,低分子肝素相对分子质量降低,抗凝活性降低,抗栓作用增强。     

亲和色谱分离纯化低分子肝素

目的探索亲和色谱纯化低分子肝素的方法。方法以壳聚糖为载体,以抗凝血酶Ⅲ及凝血酶为配基分别制备亲和吸附剂,采用亲和方法分离纯化低分子肝素,并对两者的纯化效果进行比较。结果前者制备的低分子肝素抗Ⅹa因子活性为173.9 IU/mg,比纯化前提高1.7倍;后者制备的低分子肝素抗Ⅹa因子活性为646.0 IU/mg,比纯化前提高6.3倍。结论经壳聚糖-抗凝血酶Ⅲ纯化的低分子肝素其抗Ⅹa因子活性显著提高。     

用分光光度法测定低分子肝素的效价

低分子肝素（ＬｏｗＭｏｌｅｃｕｌａｒＷｅｉｇｈｔＨｅｐａｒｉｎ,ＬＭＷＨ）比普通肝素有较强的抗活化Ⅹ因子（ＦⅩａ）作用和较弱的抗凝血酶（即活化Ⅱ因子,ＦⅡａ）的作用,抗ＦⅩａ与抗ＦⅡａ的活性比率大［１～５］,故抗血栓作用强,出血副作用小,且生物利用度...     

肝素衍生物的抗凝血活性评价

目的评估一系列化学修饰肝素衍生物的抗凝血活性。方法应用凝血仪测定肝素的部分活化凝血活酶时间(APTT)-肝素浓度的标准曲线,应用酶标仪测定肝素抗Ⅹa因子活性,制定其在405nm的吸光度-肝素浓度标准曲线,然后分别测定肝素衍生物的APTT和抗Ⅹa因子活性,根据标准曲线评价肝素衍生物的抗凝活性。结果化学修饰能够适当降低肝素衍生物的抗凝血活性,并且随着引入的有机基团的增多,肝素衍生物的抗凝血活性逐渐下降,但仍然保持一定的生物活性,且应用APTT评估肝素衍生物抗凝活性与应用抗Ⅹa因子活性评估肝素衍生物抗凝活性,测定结果基本一致。结论相比较肝素,这些化学修饰的肝素衍生物有较低的抗凝血活性,为安全应用这些肝素衍生物提供了更加广阔的前景。     

卡拉胶寡糖与bFGF的结合活性及其对乙酰肝素酶活性的抑制作用

为探讨卡拉胶寡糖作为硫酸肝素类似物的抗肿瘤和抗血管新生机制,以宫颈肿瘤细胞HeLa和人脐静脉内皮细胞HUVEC为研究对象,考察几种不同聚合范围的寡糖结合bFGF及抑制乙酰肝素酶活性的作用。发现卡拉胶寡糖对正常细胞和肿瘤细胞均表现出低毒特征。在低浓度下,具有结合bFGF并能抑制bFGF引起的细胞增殖能力,其中λ-卡拉胶寡糖(聚合度2～8)效果最明显,在质量浓度为20μg.mL-1时,可达30%的抑制率。几种寡糖对乙酰肝素酶活性有不同趋势的抑制作用,在HeLa细胞中,λ-卡拉胶寡糖的抑制活性最高;在HUVEC细胞中,聚合度在9～17的κ-卡拉胶寡糖活性最高。结果表明,卡拉胶寡糖的类硫酸肝素生物活性与其分子量大小、硫酸取代量有明显的关系,低分子量、高硫酸基取代是其高活性的关键。     

首批低分子肝素国家标准品的建立

低分子肝素 (ＬＭＷＨ )比普通肝素抗血栓作用强 ,出血副作用小 ,有较高生物利用度和较长的生物半衰期[1] ,其抗血栓作用与其抗活化Ⅹ因子(抗ＦⅩａ)活性相关 ,抗凝作用与其抗活化Ⅱ因子(抗ＦⅡａ即抗凝血酶 )活性相关[2 ,3] ,英国药典ＢＰ1998已收载该品种 ,我国已有多家单位研究生产并申报新药 ,ＬＭＷＨ质量标准拟上中国药典2 0 0 0年版增补版 ,而质量标准实施中必须要有标准品。ＷＨＯ已于 1987年制备并发行ＬＭＷＨ第一次国际标准品 ,批号 85/60 0 [4 ] ,以及 1993年发行用于测定分子量分布的ＬＭＷＨ第一次国际参考品(ＩＲＲ) ,批号 9…     

低分子量肝素治疗不稳定型心绞痛的实验室监测

目的对常规剂量低分子量肝素 (LMWH)治疗不稳定型心绞痛病人进行实验室监测。方法不稳定型心绞痛病人 2 0例 ,LMWH 5 0 0 0IU ,每日 2次皮下注射 ;测定用药前及用药后 4h病人血液中抗Ⅹa因子、活化部分凝血活酶时间 (APTT)及凝血酶时间 (TT) ,并进行统计分析。结果治疗前后病人血液中抗Ⅹa因子、APTT及TT均有显著性变化 ,常规剂量LMWH治疗不稳定型心绞痛病人时 ,不同体重的病人其抗Ⅹa因子有极显著性差异。结论常规剂量LMWH是一种安全的治疗不稳定型心绞痛的药物 ,为达到最佳的治疗效果 ,应根据不同个体 ,调整剂量 ,进行实验室监测     

低分子肝素钙治疗急性心肌梗死的安全性评价及临床疗效研究

临床流行病学调查显示,我国冠心病的发病率明显增加,急性心肌梗死(ectur emyocardial infraction,AMI)的发病年龄呈显著的下延 ,逐渐成为严重威胁人们生命的主要危险.急性心肌梗死是心肌缺血性坏死,为在冠状动脉病变的基础上,发生冠状动脉血供急剧减少或中断,使相应心肌严重而持久的急性缺血所致.低分子肝素是一种新型的抗凝血酶Ⅲ(AT-Ⅲ)依赖性抗血栓形成药,其药理作用与普通肝素基本相似.可分为抗血栓活性和抗血凝活性,血浆中凝血酶(即因子Ⅱa)活性与血凝关系密切,因子Ⅹa 活性与血栓形成关系较密切.由于本品对因子Ⅹa 活性的抑制作用强于抗凝血酶Ⅱa活性,且对血小板聚集功能等的影响较普通肝素小,较少发生出血合并症,对体内、外血栓,动、静脉血栓的形成有抑制作用,不被血小板的第4因子中和,对血小板功能无明显影响[1].其药代动力学参数优于普通肝素,皮下注射生物利用度高可达98%,而普通肝素只有30%,其血浆抗因子Ⅹa 活性3h达到血浆峰值,直至用药24h后仍可监测到,消除半衰期较长.由于低分子肝素抗因子Xa活性强,能刺激内皮细胞释放组织因子凝血途径抑制物和纤溶酶原活化物,对血栓溶解有间接协同作用,对凝血酶抑制作用较弱,对血小板功能的影响较小.临床研究显示,低分子肝素在防治血栓栓塞性疾病中效果良好[2],低分子肝素钙是低分子肝素的钙盐,我们应用低分子肝素钙作为抗凝剂治疗急性心肌梗死,未发现出血等不良事件,疗效确切,安全性高.报告如下.     

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.     

Nachweis einiger Heilmittel in der Kniegelenksynovialflüssigkeit

Zusammenfassung Mittels Gaschromatographie und Dünschichtchromatographie wiesen die Autoren 11 Substanzen nach, welche durch Injektion oder nach Verabreichung per os in die Kniegelenksynovialflüssigkeit eindrangen. In ihrer Aufstellung konnten sie eine direkte Beziehung zwischen Struktur sowie chemischphysikalischen Eigenschaften der Substanz und ihrer Fähigkeit, aus dem Blut in die Kniegelenksynovialflüssigkeit einzudringen, nicht nachweisen, außer der Tatsache, daß Substanzen mit starker Affinität zu Eiweißstoffen erst in höheren Dosen nachweisbar waren.     

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.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.