知柏地黄丸的UPLC指纹图谱研究

目的 建立知柏地黄丸UPLC指纹图谱的研究方法。方法 采用Waters Acquity BEH C₁₈色谱柱（2.1 mm×100 mm,1.7 μm）,流动相为乙腈-0.1%磷酸,梯度洗脱,检测波长236 nm,流速0.4 mL·min^-1,柱温40℃,以丹皮酚作为参照物。结果 在12 min内完成指纹图谱分析,标出12个共有峰,对6个色谱峰进行了归属,并进行了相似度评价。结论 UPLC简便可行,建立的指纹图谱可用于知柏地黄丸的质量控制。     

鼻渊净胶囊的HPLC指纹图谱研究

目的 建立鼻渊净胶囊的高效液相色谱（HPLC）指纹图谱。方法 采用Agilent SB-C₁₈（4.6 mm×250 mm,5 μm）色谱柱,乙腈-水为流动相、以1.0 ml/min流速行梯度洗脱,检测波长210 nm,柱温30 ℃,洗脱时间为80 min。采用中药色谱指纹图谱相似度评价系统（2004A版）对检测出色谱进行指纹图谱相似度评价。结果 建立了鼻渊净胶囊的HPLC指纹图谱,确定了20个共有峰,15个峰归属到各药材,其中5个峰确认了化学成分;10批样品的指纹图谱的整体相似度与对照图谱比较,均在90%以上。结论 所建立的鼻渊净胶囊指纹图谱有助于从整体上控制该制剂的质量。     

注射用五味子提取物指纹图谱研究

目的 采用高效液相色谱法建立注射用五味子提取物指纹图谱,并对2015—2016年制备的40批注射用五味子提取物进行指纹图谱相似度分析。方法 试验考察了不同流动相组成和浓度、不同检测波长、不同洗脱方法、不同品牌C₁₈色谱柱（250 mm×4.6 mm,5 μm）、不同流速、不同柱温等条件下供试品溶液的色谱行为,利用中药色谱指纹图谱相似度评价系统对试验所得色谱图进行相似度分析。结果 确定了高效液相色谱检测条件,建立了20批注射用五味子提取物的对照指纹图谱（相似度不低于0.991）,确定了12个共有峰,并对5个共有峰进行了指认,其余20批供试品与对照图谱的相似度均值为0.994。结论 本试验建立的注射用五味子提取物指纹图谱可以为其质量控制提供科学的依据,为完善注射用益气复脉（冻干）质量标准提供数据支持。     

经典名方泽泻汤UPLC指纹图谱的建立

目的 建立泽泻汤的UPLC指纹图谱。方法 采用ACQUITYUPLC^ÒBEH C₁₈色谱柱（2.1 mm×50 mm，1.7 μm），以乙腈（A）-水（B）作为流动相进行梯度洗脱，柱温35℃，流速0.3 mL·min^-1，全波长扫描，以泽泻醇B为参照峰，分析15批次泽泻汤的UPLC指纹图谱，并使用中药色谱指纹图谱相似度评价系统结合主成分分析、正交偏最小二乘法判别分析（PLS-DA）评价泽泻汤的指纹图谱。结果 在泽泻汤指纹图谱中共标定23个共有峰，其中15个化合物峰来自泽泻，8个化合物峰来自白术，指认了白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ、泽泻醇A、泽泻醇B和23-乙酰泽泻醇B 6个共有峰，其含量波动范围分别为0.028 7~0.033 1，0.029 5~0.036 6，0.012 0~0.019 4，0.102 2~0.143 9，0.469 3~0.701 2，0.425 5~0.730 8 mg·mL^-1，15批样品指纹图谱相似度为0.979~0.996。主成分分析和PLS-DA将15批样品按照泽泻的产地不同分为3类。结论 该方法快速简单、精密度高、稳定性强、重复性好，基本体现了泽泻汤的整体化学成分特征，可为泽泻汤开发和应用的质量控制提供参考。     

疏血通注射液中游离氨基酸的UPLC指纹图谱研究

目的 建立疏血通注射液中游离氨基酸的UPLC指纹图谱分析方法。方法 采用异硫氰酸苯酯（PITC）衍生化方法,使用Waters Acquity超高效液相色谱仪,以CORTECS^® C₁₈柱（150 mm×2.1 mm,1.6 μm）,乙腈-0.1%甲酸水进行梯度洗脱,体积流量为0.1 mL/min,柱温为25℃,进样量为2 μL,检测波长为254 nm。结果 33批不同批次疏血通注射液中有28个共有峰,其中有15个已知峰,分别为组氨酸（His）、精氨酸（Arg）、丝氨酸（Ser）、甘氨酸（Gly）、谷氨酸（Glu）、天冬氨酸（Asp）、苏氨酸（Thr）、脯氨酸（Pro）、丙氨酸（Ala）、酪氨酸（Tyr）、甲硫氨酸（Met）、缬氨酸（Val）、亮氨酸（Leu）、赖氨酸（Lys）、苯丙氨酸（Phe）,不同批次的样品与标准指纹图谱相似度均在0.994以上。结论 本实验建立的UPLC指纹图谱分析方法稳定、精密度高,可应用于疏血通注射液的质量控制和评价。     

泰山紫草与新疆紫草的HPLC指纹图谱对比研究

目的 对比研究泰山紫草和新疆紫草指纹图谱及其主要成分含量,评价泰山紫草的药用价值。方法 采用HPLC,色谱柱为Symmetry C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为乙腈-水（70∶30）,流速为1.0 mL·min^-1,柱温为30℃,检测波长为516 nm。应用中药色谱指纹图谱相似度评价系统（2012年版）软件,建立泰山紫草、新疆紫草指纹图谱。结果 建立了泰山紫草、新疆紫草指纹图谱的共有模式,以新疆紫草对照图谱作为参照图谱,进行峰匹配,对比计算泰山紫草指纹图谱的相似度为0.798,泰山紫草中含有萘醌类有效成分,但含量与新疆紫草中存在差异。结论 泰山紫草具有一定的药用价值,但不可完全替代新疆紫草使用。     

党参生品及不同米炒党参UPLC指纹图谱比较研究

目的 比较小米和大米炒制党参Codonopsis Radix前后的指纹图谱差异。方法 采用超高效液相色谱（UPLC）法,色谱柱为Agilent Poroshell SB-C₁₈,流动相为乙腈-0.15%冰醋酸水（梯度洗脱）,体积流量为0.3 mL/min,检测波长为267 nm,柱温为30℃,进样量为1 μL。以党参炔苷为参照,建立生党参、小米炒党参、大米炒党参（各9批）的UPLC指纹图谱;运用《中药指纹图谱相似度评价系统（2012版）》进行相似度评价及共有峰指认;运用SPSS 20.0软件进行聚类分析;使用数据统计软件SSPS 20.0与SIMCA 14.1进行主成分分析。UPLC法检测大米炒党参、小米炒党参、生党参各9批饮片中5-羟甲基糠醛（5-HMF）和党参炔苷含量。结果 9批党参生品的UPLC指纹图谱有6个共有峰,小米炒党参和大米炒党参分别均有10个共有峰,相似度均大于0.90;指认了5-HMF、党参炔苷2个共有峰,党参“米”炒后UPLC指纹图谱共有峰特征明显,小米炒党参和大米炒党参的UPLC指纹图谱未见明显差异。聚类分析和主成分分析结果一致,9批党参生品聚为一类,而小米炒党参和大米炒党参聚为另一类。与生党参比较,大米炒党参和小米炒党参中5-HMF含量均显著升高（P<0.01）,3种饮片党参炔苷含量无明显差异。结论 成功建立了党参生品及小米炒党参、大米炒党参的UPLC指纹图谱,不同炮制辅料对米炒党参中化学成分的影响趋势相同,但程度不同。     

HPLC-Q-TOF-MS技术在预知子配方颗粒指纹图谱建立和化学成分归属中的应用

目的 建立预知子配方颗粒的HPLC指纹图谱，并使用Q-TOF-MS技术鉴定共有峰成分。方法 采用Agilent Poroshell 120 EC-C₁₈色谱柱（4.6 mm×50 mm，2.7 mm），以乙腈-0.05%甲酸水溶液为流动相进行梯度洗脱，检测波长210 nm，柱温30℃，流速0.5 mL·min^-1。采用相似度分析软件对指纹图谱进行分析。以标准物质和文献数据为参照，使用Q-TOF-MS负离子检测模式对特征峰进行质谱分析并指认。结果 建立了预知子配方颗粒的HPLC指纹图谱，以4，5-O-二咖啡酰基奎宁酸峰为参照峰，标定了14个共有峰，样品间相似度范围为0.943~0.994，样品与对照指纹图谱相似度范围为0.976~0.988。使用Q-TOF-MS对共有峰进行了指认，确定了8个峰成分，分别为新绿原酸、绿原酸、木通苯乙醇苷B、3，4-O-二咖啡酰基奎宁酸、3，5-O-二咖啡酰基奎宁酸、4，5-O-二咖啡酰基奎宁酸、川续断皂苷Ⅵ、α-常春藤皂苷。结论 该方法灵敏、准确、可靠，可用于预知子配方颗粒的质量评价。     

恤彤注射液中间体丹参提取液的指纹图谱分析及多指标含量测定

目的 建立恤彤注射液中间体丹参提取液的HPLC-UV指纹图谱,并对其中的5-羟甲基糠醛、丹参素、原儿茶醛、迷迭香酸、紫草酸、丹酚酸A及丹酚酸B共7种指标成分进行定量分析。方法 采用Waters Cortecs C₁₈色谱柱（100 mm×4.6 mm,2.7 μm）,流动相0.4%甲酸水溶液（A）-0.4%甲酸乙腈（B）,流速0.8 mL·min^-1,梯度线性程序洗脱,进样量3 μL,柱温25℃,检测波长280 nm。结果 在建立的色谱条件下,指纹图谱峰型对称,分离度较好,能较全面地反映丹参提取液的化学信息。共确定了24个共有峰,10批丹参提取液的指纹图谱相似度均>0.960;在一定质量范围内,7种指标成分的峰面积与质量浓度均呈良好的线性关系,加样回收率为96.49%~100.18%,RSD为1.24%~2.27%（n=9）,所测定的7种指标成分总量为8 202.2~12 078.7 μg·g^-1。结论 建立的分析方法准确、简单、重复性好,HPLC-UV指纹图谱结合定量测定能更全面地反映丹参提取液的质量,可用于恤彤注射液中间体丹参提取液的质量评价。     

岭南特色饮片熟党参HPLC指纹图谱及其炮制前后对比研究

目的 建立同时适用于岭南特色饮片熟党参及其生品的HPLC指纹图谱，分析对比党参蒸制前后成分的变化，为进一步建立熟党参质量控制标准奠定基础。方法 Waters Symmtry C₁₈反相色谱柱（250 mm×4.6 mm，5 μm）；流动相为甲醇-0.1%甲酸水溶液，梯度洗脱（柱温30℃，流速1.0 mL·min^-1，检测波长283 nm，记录时间55 min）。使用中药色谱指纹图谱相似度评价系统（2012版）进行数据处理。结果 不同批次熟党参样品的指纹图谱相似度为0.750~0.979，存在一定的差异性，结合生、熟品色谱峰差异特点选定10个共有特征峰。结论 所建立的方法稳定、重现性好，熟党参与生品之间指纹图谱的差异对比结果为建立熟党参质控标准提供了参考依据。     

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.     

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.