耐卡铂人卵巢癌细胞相关蛋白的筛选研究

目的：筛选耐卡铂人卵巢癌细胞相关蛋白质,从蛋白质水平探讨耐药的分子机制。方法：提取人卵巢癌亲代细胞SKOV3和对卡铂耐药的细胞SKOV3/CB的总蛋白,用二维液相色谱技术分离、筛选二者之间的差异蛋白,用电喷雾离子阱质谱技术分析鉴定蛋白质。结果：共分离筛选出差异蛋白54个,其中SKOV3细胞系表达上调者20个,SKOV3/CB细胞系表达上调者34个。初步鉴定出SKOV3/CB细胞系4个表达上调的耐药相关差异蛋白,即超氧化物歧化酶1、周期蛋白依赖激酶6抑制因子、转录适配蛋白1和垂体腺苷酸环化酶促多肽。这些蛋白涉及氧化还原、细胞周期调控及凋亡等多方面。结论：人卵巢癌对卡铂耐药的机制可能与多种蛋白质有关;有关实验数据对进一步研究人卵巢癌对铂类的耐药有一定价值。     

竹叶提取物的抗血栓作用

目的从抗血小板聚集,抗凝血,及纤溶3个方面研究竹叶提取物的抗血栓作用。方法采用小鼠肺血栓模型、家兔颈总动脉血栓模型,测定家兔体外血浆凝血酶时间、凝血酶原时间、部分凝血活酶时间、血浆复钙时间和小鼠优球蛋白溶解时间及小鼠全血凝块质量等指标,考察竹叶提取物的抗血栓作用。结果竹叶提取物静脉给药可显著提高胶原蛋白肾上腺素混合诱导剂所致肺血栓小鼠的存活率;明显抑制家兔颈总动脉血栓形成;显著降低小鼠优球蛋白溶解时间,减轻小鼠全血凝块重量;其体外给药能显著延长家兔凝血酶、凝血酶原、部分凝血活酶时间,延长血浆复钙时间。结论竹叶提取物具有明显的抗血栓形成作用。     

Studies on exposure status of inhabitants to water-arsenic valence states in areas with endemic arsenism in the Datong basin in Shanxi

This study aimed to describe the distribution of water-arsenic (As) valence states and its relationship to areas with endemic arsenism in the Datong basin. Drinking water samples of patients with endemic arsenism and a control group were examined using hydride generation atomic fluorescence spectrometry (HG-AFS). We analyzed the data using SPSS10.0 for Windows. The As(III)/As ratio was 52.1% in the water sample, exceeding the national standard of 0.05 mg/L. The As(III)/As ratio significantly varied among the different stages in the disease-state groups, and with the control group (χ ² = 22.4, P<0.01). The As(III)/As(V) ratio significantly varied in the four groups (χ ² = 26.19, P<0.01), with a tendency to increase along with the seriousness of the disease state. The most common type of drinking water arsenic valence state was As(III) in the endemic disease-areas. Endemic arsenism was positively correlated with As(III). This led us to conclude that the fraction of each water-arsenic valence state should be studied when determining the arsenic content of drinking water. Translated from Chinese Journal of Endemiology, 2006, 25(1): 64–66 [译自: 中国地方病学杂志]     

Development and validation of a LC–MS/MS method for determination of bivalirudin in human plasma: Application to a clinical pharmacokinetic study

A simple, sensitive and rapid LC–MS/MS method has been developed and validated for the identification and quantification of bivalirudin in human plasma using nafarelin as the internal standard. Following protein precipitation with methanol, the analytes were separated on a C₁₈ column interfaced with a triple-quadrupole tandem mass spectrometer using positive electrospray ionization. Quantification of bivalirudin was conducted by multiple reaction monitoring (MRM) of the transitions of m/z 1091.4 → (356.4 + 227.4) for bivalirudin and m/z 662.4 → 328.5 for IS. The lower limit of quantification was 1.25 ng/ml, and the assay exhibited a linear range of 1.25–500 ng/ml. The developed assay method was successfully applied to a pharmacokinetic (PK) study in healthy volunteers after intravenous administration of bivalirudin.     

基于数学模型的非洛地平缓释片检验指标综合评价的探讨

目的为客观评价合格药品的生产质量状况,有效区分其存在的差异程度,通过建立一定的数学模型,对特定药品每一项可定量的检验指标进行综合评价。方法本数学模型将药品的检验指标参数以理论最佳点做理想值,以药品合格标准做下限值,转化成各指标的评价指数,并使各评价指数同量齐次;各检验指标的评价指数经过带权重的理想点法计算,得到综合评价结果;其中,运用专家经验评定法确定各检验指标的权重。结果数学模型确立后,尝试以非洛地平缓释片各项检验数据为例进行试评价,获得了较为符合客观实际的结果,说明了用数学模型评价药品检验的各项指标的可行性和可操作性,该方法也可以推广至其他药品检验指标的综合结果判断。结论利用数学模型对药品检验结果进行评价的方法,可以成为药品质量监督管理的新思路,也可以为药品按质定价提供参考性依据。     

Dry powder inhalations containing thymopentin and its immunomodulating effects in Wistar rats

Thymopentin (TP5), a synthetic pentapeptide, has been used in clinic as a modulator for immunodeficiences through intramuscular administration. The purpose of this study was to design and evaluate dry powder inhalations (DPIs) for pulmonary delivery of TP5. Dry powder inhalations containing leucine (a dispersibility enhancer), mannitol, and lactose (bulking agents) were prepared by spray-drying from aqueous formulations. The formulation components on the aerosolisation characteristics of spray-dried powders were investigated through the use of various amount of leucine, lactose and mannitol. Following spray-drying, resultant powders were characterized using scanning electron microscopy, laser diffraction and tapped density measurements, and the aerosolisation performance was determined using Twin Stage Impinger. The immunosuppression Wistar rats model was constructed to evaluate the immunomodulating effects of TP5 DPIs in vivo. The results of T-lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+ ratio) analyses suggest that TP5 DPIs have modulating effects. On an overall evaluation, TP5 pulmonary delivery DPIs may be feasible for the future clinical application.     

短程小剂量口服激素在婴幼儿喘息急性期联合降阶梯治疗方案中的作用研究

目的：前期研究表明,与目前常用的静脉激素及抗生素治疗方案相比,由泼尼松＋阿奇霉素＋妥洛特罗贴剂＋氯雷他定＋孟鲁司特钠组成的联合降阶梯治疗方案对婴幼儿喘息具有更好的疗效。在此基础上,本研究将进一步探讨作为联合降阶梯治疗方案组分之一的泼尼松在其中的作用。方法：选取2011年10—12月来我院哮喘门诊就诊的婴幼儿喘息患儿,共计100例,随机分为泼尼松组和对照组各50例。泼尼松组用药方案：泼尼松0．5mg／（kg·d）×3d,每天1次晨顿服;阿奇霉素10mg／（kg·d）×3d,每天1次口服;妥洛特罗贴剂0．5mg／d（1贴）×7d;氯雷他定糖浆3mL／d×14d,每天1次口服;孟鲁司特钠4mg／d×14d,每天1次口服。对照组除无泼尼松外,其他药物用法用量同泼尼松组。结果：（1）两组在治疗第3天和第7天咳嗽、喘息、哮呜音症状均较治疗前好转。（2）泼尼松组在治疗第3天咳嗽、哮呜音症状评分以及临床疗效均优于对照组（P〈0．05）。结论：短程小剂量口服激素是婴幼儿喘息急性期联合降阶梯治疗方案的有效组分之一。     

基于OSMAC策略对深海来源真菌Acrostalagmus luteoalbus SCSIO F457化学多样性的初步研究

摘要：目的 采用单菌多次级代谢产物（OSMAC）策略对1株采自南海深海2 801 m沉积环境的白黄笋顶孢霉属真菌Acrostalagmus luteoalbus SCSIO F457进行化学多样性的初步研究。方法 通过在不同培养基、不同pH与不同盐度条件下对菌株进行培养调控并筛选2种适宜发酵条件进行小规模发酵。采用硅胶柱层析、葡聚糖凝胶层析、半制备高效液相等色谱学方法对发酵产物进行化学分离,利用NMR、MS等波谱学技术并结合文献鉴定化合物结构,并对化合物进行初步抗氧化和抗菌活性测试。结果 从菌株SCSIO F457的发酵产物中共新增分离鉴定11个单体化合物,包括paulownin（1）、cyclo(L-Phe-L-Pro)（2）、cyclo(L-Tyr-L-Pro)（3）、cyclo(L-Val-L-Pro)（4）、cyclo(D-Ile-L-Pro)（5）、cyclo(D-Leu-L-Pro)（6）、1-methyoxy-4-(2-hydroxy)ethylbenzene（7）、2-(4-hydroxyphenyl)-ethanol（8）、1-phenylbutane-2,3-diol（9）、3-methoxy-2-methyl-4H-pyran-4-one（10）及3-(hydroxy-acetyl)-1H-indole（11）,化合物7表现出较弱的1,1-二苯基-2-苦基肼（1,1-Diphenyl-2-picryl-hydrazyl, DPPH）自由基清除活性。     

转运体在药物经肝脏清除过程中的作用

肝脏在药物的体内清除过程中具有重要作用,它不仅是药物代谢的主要场所,还控制着药物及其代谢物的胆汁排泄过程。转运体是控制细胞内外物质传输的一类功能性膜蛋白,其在肝脏有广泛表达,并能对药物进入肝细胞以及排泄至胆汁的过程进行调控,因而,对于肝脏清除过程具有重要作用。本文从肝脏中重要转运体的分布、功能以及底物选择性出发,对其在药物的肝脏清除中的作用、由其引起的药物药物相互作用以及重要转运体的基因多态性研究进行了综述。     

Tanshinone IIA triggers p53 responses and apoptosis by RNA polymerase II upon DNA minor groove binding

Our previous work has shown that tanshinone IIA (Tan IIA) is a DNA minor groove binder instead of an intercalator as previously thought. In this study, we have further demonstrated that the molecular antitumor pharmacology of Tan IIA is dependent on its groove-binding capability. First, we investigated the structure damage to duplex DNA upon Tan IIA binding using circular dichroism spectra. Subsequently, we performed western blot, flow cytometry analysis, chromatin immunoprecipitation, and quantitative real-time PCR to illustrate the RNAPII degradation, phosphorylation, and distribution along the transcribed gene in H22 cells exposed to Tan IIA. In addition, p53 activation and apoptosis induction in both cultured H22 cells and in mice bearing the ascitic-type H22 were measured following Tan IIA treatment. It was revealed that Tan IIA decreases the level of RNAPII by altering DNA structure. At the low dose range (0.2-4 μM) of Tan IIA exposure, the DNA structure damage results in the inhibition of RNAPII binding to DNA and the initiation of RNAPII phosphorylation, while higher concentrations of Tan IIA (4-20 μM) cause complete phosphorylation and degradation of RNAPII followed by p53 activation and apoptosis. A similar apoptosis induction by RNAPII was observed in animals. Apoptosis of tumor cells from ascitic fluid was not detected until RNAPII levels were downregulated by Tan IIA, which requires 40 mg/kg body weight of Tan IIA. It was concluded that DNA-conformational-damage-dependent RNAPII response upon groove binding is the molecular basis of the antitumor property of Tan IIA, in vivo and in vitro.