Immunomodulatory and cytotoxic effects of Nigella sativa and thymoquinone on rat splenocytes

Three different concentrations of Nigella sativa (N. sativa) ethanolic extract, thymoquinone (TQ), dexamethasone, and saline were examined to see whether they had any effects on cell viability, proliferation, and interleukin 4 (IL-4) and interferon-γ (IFN-γ) secretion in non-stimulated, phytohemagglutinin (PHA) and concavaline A (Con A)-stimulated splenocytes. In PHA and Con A-stimulated splenocytes, cell viability and proliferation were increased and Con A shifted cytokine profile towards Th2 balance. Dexamethasone treatment showed a suppression in viability, IFNγ and IL-4 secretion in non-stimulated and stimulated splenocytes. Extract and TQ reduced the viability and inhibited the proliferation of stimulated and non-stimulated splenocytes concentration-dependently. Higher concentrations of N. sativa (1000 mg/ml) and TQ (5 and 10 mg/ml) reduced the secretion of IL-4 in stimulated cells. Two higher concentrations of N. sativa had decreased IFNγ secretion in both stimulated and non-stimulated cells. In non-stimulated cells, only the highest and in Con A-stimulated cells, all TQ concentrations had inhibited IFNγ secretion. The highest concentration of N. sativa increased IFNγ/IL-4 ratio in both stimulated and non-stimulated cells while higher concentrations of TQ only had the same effect on stimulated cells. N. sativa and TQ showed cytotoxic inhibitory effect on rat splenocytes and on Th1/Th2 cytokines concentration-dependently. Higher concentrations of extract and TQ increased cytokines balance in Th1/Th2.     

拳参的DNA裂解活性成分研究

目的：研究蓼科植物拳参的水溶性成分，并测试其DNA裂解活性。方法：利用反相层析的方法进行分离纯化，根据化合物的化学性质与光谱数据鉴定其结构。结果：自拳参根茎的60％丙酮提取物中分得10个化合物，确定其结构分别为没食子酸（I），色氨酸（Ⅱ），2，6－二羟基苯甲酸（Ⅲ），（＋）－儿茶素（Ⅳ），绿原酸（Ⅴ），（－）-表儿茶素5－O－β－D－吡喃葡萄糖苷（Ⅵ），（＋）－儿茶素－7－O－β－D－吡喃葡萄糖苷（Ⅶ），1－（3－O－β－D－吡喃葡萄糖革－4，5－二羟基－苯基）－乙酮（Ⅷ），（＋）-儿茶素－5－O－β－D－吡喃葡萄糖苷（Ⅸ）和（－）-表儿茶素（Ⅹ）。结论：化合物Ⅱ，Ⅲ，Ⅴ-Ⅹ为首次从该植物中分得，化合物Ⅰ，Ⅳ，Ⅵ，Ⅶ，Ⅸ，Ⅹ具有很强的DNA裂解活性。     

近红外漫反射法测定茯苓中水溶性多糖及碱溶性多糖

目的:建立茯苓中水溶性多糖和碱溶性多糖的基于近红外光谱的含量预测模型。方法:建立并优化茯苓中水溶性多糖和碱溶性多糖的分光光度含量测定方法,并用建立的分光光度法对90个批次样品中的碱溶性多糖和水溶性多糖进行含量测定。采用积分球漫反射对样品进行红外光谱扫描,参考分光光度法获得的2种多糖的含量结果,经光谱预处理方法优选,并以Matlab R2012a软件,采用组合区间偏最小二乘法(synergy interval partial least squares regression SIPLS)对特征波长进行了筛选,建立了茯苓中水溶性多糖和碱溶性多糖的近红外含量测定方法。结果:建立的近红外预测模型水溶性多糖交叉验证均方根误差(RMSEC)为0.270,相关系数R_c~2=0.905 9,预测均方根误差(RMSEP)为0.119;预测集相关系数R_p~2=0.913 8。碱溶性多糖RMSEC=0.242,Rc2=0.999 6;RMSEP=0.243,Rp2=0.998 9。结论:所建立的近红外漫反射含量测定方法操作简便,快速,准确。适于茯苓中水溶性多糖和碱溶性多糖的质量控制。     

Physical–chemical properties of furosemide nanocrystals developed using rotation revolution mixer

Recently, several approaches have been reported to improve the dissolution rate and bioavailability of furosemide, a class IV drug. However, to the best of our knowledge, none of them proposed nanocrystals. In the last decade, nanocrystals successfully addressed solubility issues by increasing surface area and saturation solubility, both leading to an increase in the dissolution rate of poor water soluble drugs. The preparation of furosemide nanocrystals was by a rotation revolution mixer method. Size distribution and morphology were performed using laser diffraction and scanning electron microscopy, respectively. In addition, differential scanning calorimetry, thermogravimetry, X-ray powder diffraction (XRD) and low frequency shift-Raman spectroscopy allowed investigating the thermal properties and crystalline state. Solubility saturation and intrinsic dissolution rate (IDR) studies were conducted. The thermal analysis revealed lower melting range for the nanocrystals comparing to furosemide. Moreover, a slight crystalline structure change to the amorphous state was observed by XRD and confirmed by low frequency shift Raman. The particle size was reduced to 231?nm with a polydispersity index of 0.232, a 30-fold reduction from the original powder. Finally, the saturation solubility and IDR showed a significant increase. Furosemide nanocrystals showed potential for development of innovative formulations as an alternative to the commercial products.     

Enzymatic Method for Assaying Uric Acid in Serum with a New Tetrazolium Salt Produces Water-Soluble Formazan Dye

Objectives: To apply an enzymatic method for assaying uric acid in serum based on the uricase (EC 1.7.3.3)-catalase (EC 1.11.1.6)-formaldehyde dehydrogenase (FADH, EC 1.2.1.46) coupled with the new tetrazolium salt producing a water-soluble formazan dye as an indicator system. Unlike the traditional tetrazolium salts, e.g., iodonitrotetrazolium (INT) and nitrotetrazolium blue (NTB), the corresponding formazan dye produced did not absorb to the test tube and the reaction cells of the analyzer. Moreover, this formazan dye had a higher water solubility and more sensitivity.

Design and Methods: A two electron reduction of tetrazolium salt with NADH, produced by the uricase-catalase-FADH reaction, was mediated by an electron carrier, 1-methoxy PMS. The generation of formazan, monitored at 440 nm, was proportional to the concentration of uric acid in serum. The sensitivity of this method was about 1.5 times that of the peroxidase-TOOS [N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine] method. The assay was evaluated with TBA-80FR · NEO biochemical analyzer. The average within-run and between-day imprecision (CV) were 0.75–2.44% and 3.20–3.33%, respectively.

Results: Results of the proposed method (y) correlated well with those determined by the conventional chromogen method (x): y = 0.970 x − 0.018 mmol/L (Sy | x = 0.019 mmol/L, r = 0.993, n = 67).

Conclusions: We also present data showing that the method is rapid, relatively free of interference, and amenable to automation.     

羧基水溶性量子点在鼻咽癌标志物EBNA1标记中的应用

目的探讨605nm羧基水溶性量子点在鼻咽癌标志物EB病毒核抗原1(EBNA1)中的标记方法。方法将605nm羧基水溶性量子点和鼻咽癌EBNA1在1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐的作用下共价交联,生成量子点标记的抗原,对标记后的抗原采用紫外可见吸收谱、荧光光谱测定,琼脂糖凝胶电泳分析。结果羧基水溶性量子点与鼻咽癌EBNA1抗原通过表面氨基与羧基缩合形成的稳定共价键,羧基水溶性量子点与鼻咽癌EBNA1抗原之间实现了成功连接,标记后荧光发射光谱检测结果表明,标记后的量子点EBNA1溶液的发射峰位置在380nm附近,保持良好的荧光特性。结论 605nm羧基水溶性量子点采用共价交联法可稳定标记鼻咽癌标志物EBNA1,为后续研究提供可靠依据。     

水溶性柠檬苦素苯甲酰腙衍生物的合成及其抗菌活性

柠檬苦素是一种在柑橘属植物中含量较丰富的化合物,具有较广泛的生物活性,但因其水溶性较差,限制了其实际应用。本研究以柠檬苦素为起始原料,在其7位羰基处引入苯甲酰腙,设计、合成了8个新化合物7a～7h。实验结果表明,所有化合物的水溶性较柠檬苦素均有所提升,其中化合物7a、7d、7e和7f较为明显;此外,在苯环间位和对位上有取代羟基的化合物7d和7e对大肠埃希菌（E. coli）和金黄色葡萄球菌（S. aureus）具有较强的抗菌活性,化合物7e活性最好,其最低抑菌浓度分别为0.31 mg/mL和1.25 mg/mL,且化合物7e的抗E. coli效果优于苯甲酸钠。     

水溶性脂质体运载基因药物复合物治疗大鼠神经病理性痛

背景：有研究报道病毒载体运载 N-甲基-D 天冬氨酸受体1小干扰 RNA 可有效缓解大鼠炎性疼痛,但病毒载体存在安全隐患.目的：探讨水溶性脂质体运载 N-甲基-D 天冬氨酸受体1小干扰 RNA 在体内外沉默 N-甲基-D 天冬氨酸受体1的效应和治疗神经病理性痛的可行性.方法：将 PC12随机分为阴性转染组、对照转染组和水溶性脂质体转染组,分别以 N-甲基-D-天冬氨酸受体1小干扰 RNA、聚乙烯亚胺与 N-甲基-D-天冬氨酸受体1小干扰 RNA 的复合物及水溶性脂质体与 N-甲基-D-天冬氨酸受体1小干扰 RNA 的复合物转染 PC12细胞,检测各组 N-甲基-D-天冬氨酸受体1基因 mRNA 及蛋白水平表达的变化.将48只 SD 大鼠随机分为假手术组、模型组、聚乙烯亚胺组及水溶性脂质体组,后3组建立大鼠神经病理性疼痛模型,并分别鞘内注射生理盐水、聚乙烯亚胺与N-甲基-D-天冬氨酸受体1小干扰 RNA 的复合物和水溶性脂质体与 N-甲基-D-天冬氨酸受体1小干扰 RNA 的复合物;假手术组只暴露坐骨神经.结果与结论：水溶性脂质体转染组 N-甲基-D-天冬氨酸受体1的 mRNA 与蛋白表达水平明显低于其他两组（P 〈0.01）.与假手术组比较,模型组、聚乙烯亚胺组及水溶性脂质体组 N-甲基-D-天冬氨酸受体1的 mRNA 和蛋白表达上调,累积疼痛评分升高（P 〈0.01）;与模型组比较,水溶性脂质体转染组脊髓背角 N-甲基-D-天冬氨酸受体1 mRNA 与蛋白表达及累积疼痛评分下降（P 〈0.01）,聚乙烯亚胺组上述指标无明显变化（P 〉0.05）.表明在体内条件下水溶性脂质体可有效运载 N-甲基-D-天冬氨酸受体1小干扰 RNA,抑制 N-甲基-D-天冬氨酸受体1的过度表达,还可减轻大鼠神经病理性痛.     

四种不同类型甲壳胺抑瘤作用的初步比较

目的制备并表征纳米级甲壳胺,初步比较其与O-羧甲基甲壳胺、水溶性低分子量甲壳胺、非水溶性高分子量甲壳胺对人结肠癌LoVo细胞的抑瘤作用。方法采用改进的离子凝胶法制备纳米级甲壳胺,通过透射电子显微镜、马尔文激光纳米粒度测定仪对其形态、粒径、表面电位进行表征。采用MTT法初步比较上述四种类型甲壳胺对人结肠癌LoVo细胞的抑瘤活性。结果制备得到平均粒径160nm的纳米级甲壳胺,高、中浓度下均对LoVo细胞生长有一定的抑制作用,高浓度的抑制率达54.0%,其它三种类型甲壳胺抑制率分别是O-羧甲基甲壳胺4.5%、非水溶性甲壳胺35.8%和低分子量甲壳胺38.1%。结论对人结肠癌LoVo细胞,四种类型甲壳胺中纳米级甲壳胺的体外抑瘤作用最好,O-羧甲基甲壳胺几乎无抑瘤作用。