HPLC／DAD／MS法同时测定苗药头花蓼中3种有效成分的含量

目的：建立HPLC法测定头花蓼中没食子酸、davidiin和槲皮苷的含量。方法：采用Alltima C18色谱柱（150mm×4．6mm，5μm），以A：2．5％醋酸水溶液，B：水-四氢呋喃-甲醇（40：10：50）为流动相进行梯度洗脱，梯度条件如下：0min，80：20；15min，10：90；20min，10：90；25min，80：20；30min，80：20。流速为0．6mL·min^-1，紫外检测波长为270nm。离子源APCI，扫描模式负离子。结果：没食子酸、davidiin和槲皮苷的线性范围（n=6）分别为0．017～2．28μg（r=0．9999），0．05～2．50μg（r=0．9999），0．16～5．10μg（r=0．9999）；平均回收率（n=9）分别为98．0％，99．6％，98．5％。提取方法为80％乙醇冷浸12h后超声30min。结论：本方法简便、准确，可为评价不同产地的头花蓼质量提供依据。     

头花蓼咀嚼片的制备工艺研究

目的研究头花蓼咀嚼片的制备工艺。方法以没食子酸的含量、出膏率为指标,在热淋清颗粒制备的基础上,对水用量及辅料进行考察,以确定头花蓼咀嚼片的最佳制备工艺。结果头花蓼咀嚼片的处方及最佳工艺条件为,8倍量水提2次,每次1.5 h;80℃减压干燥;每个处方10 g甜菊素。结论头花蓼咀嚼片制备工艺稳定、可靠,且性状、片重差异及微生物限度考察均符合《中国药典》要求。     

头花蓼有效组分中没食子酸、原儿茶酸在模式生物斑马鱼中的代谢研究

目的采用模式生物斑马鱼研究头花蓼有效组分中没食子酸、原儿茶酸的代谢产物及其代谢转化形式,探讨模式生物斑马鱼用于研究药物代谢的可行性及合理性。方法采用UHPLC-Q-TOF/MS联用技术,结合Metabolite Tools~(TM)、质量亏损过滤(MDF)等代谢产物分析技术,对各成分暴露于模式生物斑马鱼24 h后的药液以及模式生物斑马鱼体内的代谢产物进行筛查分析。结果没食子酸、原儿茶酸经斑马鱼作用后,主要以甲基化、硫酸化反应为主。在斑马鱼体内外药液检测到没食子酸原形成分和2个甲基硫酸化产物,以及原儿茶酸原形成分、2个甲基硫酸化产物、2个硫酸化产物。结论头花蓼有效组分中没食子酸、原儿茶酸经斑马鱼代谢后存在Ⅱ相代谢产物,这与大鼠体内的代谢机制高度一致,提示斑马鱼对头花蓼有效组分的代谢具有合理性,为阐明该药的药效物质基础提供了实验依据。     

苗药热淋清颗粒原料头花蓼指纹图谱成功建立

贵州威门药业股份有限公司(简称威门药业)委托中国医学科学院北京药物研究所完成了头花蓼指纹图谱的研究工作。该研究采用国际公认的控制中药或天然药物最有效的手段———指纹图谱,对同产地和不同产地的头花蓼进行指纹图谱研究,对其共有峰和非共有峰面积进行了标定和确定,探索出了一套简便、快速、稳定、可靠的头花蓼指纹图谱测定方法。利用高效液相色谱(HPLC)建立的“头花蓼”指纹图谱,真正做到了从源头控制产品质量,保证了头花蓼单方制剂热淋清颗粒质量的稳定、可控,从而为头花蓼持续发展提供了技术保障,为热淋清颗粒走中药现代化之路…     

不同产地铁苋菜和绒毛龙芽草中没食子酸的含量测定

目的:测定不同产地铁苋菜和绒毛龙芽草中没食子酸的含量,并考察不同提取溶剂对没食子酸含量测定结果的影响.方法:采用HPLC法,Attima C18柱(5μm,4.6 mm×250 mm),流动相为乙腈-0.1%三乙胺(稀磷酸调pH值至3.5)(1.5:98.5),流速1.0 ml·min-1,检测波长212 nm.结果:线性回归方程为y=5.09×104 1.77×106X,r=0.999 8,线性范围为0.830 4～4.152 0μg.6个不同产地药材,以江西产铁苋菜和绒毛龙芽草中没食子酸含量最高,分别为0.545%和0.062%.不同提取溶剂比较,以水为提取溶剂所测铁苋菜和绒毛龙芽草中没食子酸含量最高,分别为0.465%和0.036%.结论:不同产地铁苋菜和绒毛龙芽草中没食子酸含量存在差异.     

RP-HPLC测定不同产地和药用部位景天三七中的没食子酸

目的 采用RP-HPLC法测定景天三七不同产地和药用部位中没食子酸的含量.方法 采用Kromasil C18色谱柱(250mm×4.6 mm,5μm),流动相为甲醇-5%醋酸溶液(5;95),流速1 ml·min-1,柱温30℃,检测波长274 nm.结果 没食子酸0.2026～1.2056μg与峰面积的线性关系良好;同归方程为;Y=3.627 × 106X+7.089×105(r=0.9998,n=6);平均回收率为99.5%,RSD=0.73%(n=9).结论 所建方法 简便、准确、重复性好,可用于控制民族药景天三七药材的质量.     

基于网络药理学的头花蓼中槲皮素、没食子酸和槲皮苷对幽门螺杆菌胃炎以及胃癌治疗机制研究

目的 基于网络药理学的方法研究头花蓼对幽门螺杆菌胃炎以及胃癌的药理学机制，其中槲皮素、没食子酸和槲皮苷是头花蓼的主要活性成分。方法 通过PubChem、STITCH、SEA等8个数据库获取相关数据，并使用Cytoscape软件绘制蛋白-蛋白相互作用及头花蓼-化合物-预测靶点网络图。利用DAVID数据库对相关靶点进行GO和KEGG的富集分析，并应用AutoDock软件进行分子对接，以分析hub基因与其对应小分子的结合能。结果 网络药理学分析结果表明头花蓼对幽门螺杆菌胃炎以及胃癌的作用靶点共有27个，其中degree值前6的核心靶点和其相对应的化合物分子对接的结合能均小于0，且80%的最低结合能结果≤-5.0 kJ/mol。GO和KEGG的富集结果表明头花蓼可以调控炎症反应、细胞凋亡信号通路、TNF信号通路等生物学过程，能通过影响受体复合物、蛋白质异二聚体活性发挥作用。结论 头花蓼多靶点、多途径地作用于幽门螺杆菌胃炎以及胃癌，为头花蓼老药新用提供了新的依据。     

头花蓼对胃黏膜上皮细胞GES-1细胞毒性的研究

目的：探讨头花蓼对胃黏膜上皮细胞株GES‐1增殖的影响。方法体外培养人胃黏膜上皮细胞株GES‐1,采用MTT法连续监测7d,以存活细胞数OD值对培养时间（h）作图,即得生长曲线。不同浓度头花蓼（1024μg／mL、512μg／mL、256μg／mL、128μg／mL、64μg／mL、32μg／mL、16μg／mL、8μg／mL,）与对数期GES‐1细胞共同培养48h,采用MTT法检测细胞增殖。结果（1）体外培养胃黏膜上皮细胞株GES‐1,生长曲线显示：细胞于培养的48h进入对数生长期,120h进入平台期。本实验选96h加药进行后续实验;（2）MTT实验表明：低于128μg／mL时,活细胞数量没有明显变化,细胞增殖无影响。结论头花蓼浓度高于128μg／mL时,对胃黏膜上皮细胞株GES‐1的生长有明显的抑制作用,头花蓼因细胞毒性较低具有潜在的临床应用前景,为头花蓼对H．pylori相关性胃炎的抗菌、抗炎机制奠定了实验基础。     

头花蓼对幽门螺旋杆菌相关性胃炎的细胞间隙连接通讯功能的影响

目的探讨头花蓼对幽门螺旋杆菌相关性胃炎的胃黏膜上皮细胞株-GES-1细胞间隙连接通讯功能的影响。方法采用划痕标记荧光染料示踪技术检测不同比例(幽门螺旋杆菌与细胞之比为25∶1,50∶1,100∶1)的幽门螺旋杆菌数对胃黏膜上皮细胞株-GES-1细胞作用24h后间隙连接通讯功能的影响。构建幽门螺旋杆菌相关性胃炎(Helicobacter pylori-associated gastritis,HPAG)细胞模型基础上,采用划痕标记荧光染料示踪技术检测不同浓度的头花蓼(8μg/mL、16μg/mL、32μg/mL、64μg/mL)作用24h后对幽门螺旋杆菌相关性胃炎的细胞间隙连接通讯功能的影响。结果 (1)通过划痕标记荧光染料示踪技术检测结果表明细菌与细胞之比为50∶1作用24h时,减弱了胃黏膜上皮细胞株-GES-1细胞间隙连接通讯功能(P<0.05);(2)通过划痕标记荧光染料示踪技术检测结果表明头花蓼(64μg/mL)作用24h改善了幽门螺旋杆菌相关性胃炎细胞模型的细胞连接通讯功能(P<0.05)。结论头花蓼(64μg/mL)作用24h改善了幽门螺旋杆菌相关性胃炎细胞模型的细胞连接通讯功能,为头花蓼对H.pylori相关性胃炎的抗菌、抗炎机制奠定了实验基础。     

毛果算盘子提取工艺及没食子酸含量测定

目的建立测定广西不同产地毛果算盘子中没食子酸含量的高效液相色谱(HPLC)法。方法采用正交实验优化毛果算盘子的提取工艺,用HPLC法测定毛果算盘子中没食子酸的含量。色谱柱为Hypersil C18柱(250 mm×4.6 mm,5μm),流动相为甲醇-0.2%磷酸溶液(10∶90),检测波长为270 nm,流速1.0 m L·min-1,柱温30℃。结果没食子酸在0.031~0.186μg呈良好的线性关系(r=0.999 9),平均加样回收率为102.81%,RSD为2.05%(n=9)。结论所建立的方法简单、稳定、重复性好,可用于毛果算盘子药材的质量控制。     

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.