HPLC分析不同来源藏药当佐中4种指标成分的含量

目的:建立HPLC同时测定藏药当佐中没食子酸、羟基红花黄色素-A、桂皮醛、胡椒碱含量的方法,对不同来源样品进行测定,以建立当佐的质量控制标准.方法:采用Waters XTerra RP-C18(4.6 mm×250 mm,5 μm)色谱柱,以甲醇-水(含0.1%冰乙酸)为流动相,梯度洗脱(0-22.5 min,流动相比例5:95-50:50;22.5-40 min,流动相比例50:50-80:20),流速为1.0 mL·min-1,检测波长为270 nm.结果:在此色谱条件下,没食子酸、羟基红花黄色素-A、桂皮醛、胡椒碱达到基线分离,且线性良好,线性范围分别为0.040-0.640μg(r=0.999 8),0.090-1.440 μg(r=0.999 9),0.031-0.500μg(r=0.999 9),0.092-1.477μg(r=0.998 9),平均回收率(n=6)分别为97.42%(RSD 1.9%),97.55%(RSD 2.9%).98.69%(RSD 0.96%),96.72%(RSD4.0%);不同来源当佐中该4种成分的含量变化分别为0.113-1.69,0.889-1.51.0.000-0.606,1.96-2.75 mg·g-1.结论:本研究所建立的方法,能够准确快速地测定当佐中指标成分的含量,可为该药的全面质量评价提供参考.     

Chronic ethanol consumption impairs cellular immune responses against HCV NS5 protein due to dendritic cell dysfunction

BACKGROUND & AIMS: Alcoholic patients with and without chronic liver disease have a high incidence of infection with hepatitis C virus (HCV). Long-term ethanol consumption in mice has been associated with a strikingly reduced CD8(+) cytotoxic T-lymphocyte (CTL) response to HCV nonstructural proteins following DNA-based immunization. This study evaluated the effect of ethanol on dendritic cells (DCs) as a mechanism(s) for reduced CTL activity. METHODS: Mice were fed an ethanol-containing or isocaloric pair-fed control diet for 8 weeks, followed by DC isolation from the spleen. DCs were evaluated with respect to endocytosis properties, cell surface markers, allostimulatory activity, and cytokine production following stimulation. Immune responses to HCV NS5 protein were generated by genetic immunization. Syngeneic transfer was used to determine if DC dysfunction contributed to abnormal cellular immune responses. RESULTS: Long-term ethanol exposure resulted in a reduced number of splenic DCs but did not alter endocytosis capacity. There was an increase in the myeloid and a reduction in the lymphoid DC population. Ethanol reduced expression of CD40 and CD86 costimulatory molecules on resting DCs, which was corrected following stimulation with lipopolysaccharide or poly I:C. There was impaired allostimulatory activity. Cytokine profiles of DCs isolated from ethanol-fed mice were characterized by enhanced interleukin (IL)-1beta and IL-10 and decreased tumor necrosis factor alpha, IL-12, interferon gamma, and IL-6 secretion. Impaired CTL responses to NS5 were corrected by syngeneic transfer of control DCs. CONCLUSIONS: Altered DC function is one of the major changes induced by long-term ethanol consumption, which subsequently impairs the cellular immune response necessary for viral clearance.     

应用四黄降糖丸对糖尿病高危人群预防的研究

目的：回顾性研究中药组方四黄降糖丸对糖尿病高危人群的预防效果。方法：2007年1月-2009年12月住院及查体中心确认属于糖尿病高危人群者接受生活方式干预,生活方式干预和中药组方四黄降糖丸的药物干预各100人。监测血糖、体重指数、血脂以及其并发症的发生情况,对两组的预防效果差异进行对比分析。结果：应用生活方式干预和中药处方干预的高危人群的血糖、体重指数、血脂、及其并发症的发生情况的预防效果优于单纯生活干预的预防效果。结论：应用中药组方四黄降糖丸能有效预防糖尿病高危人群的病情进展。     

益气养阴活血中药治疗慢性心力衰竭急性加重36例临床疗效观察

[目的]评价参麦注射液合注射用红花黄色素治疗慢性心力衰竭急性加重期气阴两虚证的临床疗效及安全性。[方法]将72例慢性心力衰竭急性加重期气阴两虚证患者随机分为治疗组和对照组,两组均给予西药规范治疗,治疗组在西药规范治疗的基础上联合参麦注射液合注射用红花黄色素静脉滴注,疗程10 d。[结果]治疗组在中医证候方面显著改善,且明显优于对照组(P0.05或P0.01);心功能改善情况与对照组比较差异有统计学意义(P0.05);脑钠肽(BNP)显著降低,与对照组比较差异有统计学意义(P0.05)。[结论]在西药规范治疗基础上,联合参麦注射液合注射用红花黄色素治疗慢性心力衰竭急性加重期临床疗效确切,且具有较好的安全性。     

七味红花殊胜丸质量标准研究

目的：建立七味红花殊胜丸质量标准。方法：采用薄层色谱法（TLC）对处方中红花进行定性鉴别;用高效液相色谱法（HPLC）测定羟基红花黄色素A含量。结果：TLC定性鉴别分离效果好,斑点显色清晰。羟基红花黄色素A的进样浓度在13.18~79.10μg·ml-1范围内与峰面积积分值呈良好线性关系（r=0.9998）;平均回收率为98.2%,RSD=1.27%（n=6）;每丸含红花以羟基红花黄色素A计应不少于1.0mg。结论：标准可用于该制剂的质量控制。     

红花转录因子CtMYB1基因的克隆及原核表达

目的 从红花Carthamus tinctorius 花瓣中克隆转录因子基因CtMYB1,并进行序列分析和原核表达载体的构建。方法 根据红花转录组测序结果中得到的高表达的Unigene123933序列,利用RT-PCR和RACE技术从红花花瓣中扩增得到CtMYB1基因的全长cDNA序列,并进行生物信息学分析;以CtMYB1基因的全长cDNA序列为模板,PCR扩增得到CtMYB1基因的开放阅读框(ORF)序列,构建原核表达载体pEASY-E1-CtMYB1。转化大肠杆菌BL21(DE3)进行诱导表达。结果 成功从红花中克隆1个MYB基因,命名为CtMYB1,GenBank登录号为KJ524853。CtMYB1基因全长893 bp,ORF为750 bp,编码249个氨基酸。同时,成功构建了该基因的原核表达载体。SDS-PAGE结果显示该蛋白的大小约30 000,与预测的蛋白相对分子质量一致。结论 成功克隆了CtMYB1基因,构建了原核表达载体,初步证明该基因在大肠杆菌中成功表达。     

栀子黄色素的分离纯化

目的研究栀子黄色素的大孔吸附树脂富集纯化工艺。方法以栀子黄色素的纯度和得率为指标,考察大孔吸附树脂FU02对栀子提取液动态富集纯化最适工艺。结果大孔吸附树脂FU02对栀子黄色素的最适动态吸附条件：料液中栀子黄色素浓度5g/L、栀子苷浓度30~32g/L,料液流速1Bv/h,此条件下大孔吸附树脂FU02对栀子黄色素的吸附率为82.5%,栀子苷去除率可达69.7%。吸附在树脂上的栀子黄色素的最适动态解吸条件：以80%乙醇溶液为解吸液、解吸流速2Bv/h,此条件下栀子黄色素洗脱率可达96.0%,制备的栀子黄色素A238/A440可降至0.35~0.38。结论采用大孔吸附树脂FU02富集纯化,制备高纯度的栀子黄色素,是方便可行的。     

高效液相色谱法测定蛋糕中的栀子黄

栀子甙是栀子黄的主要成分,因此以栀子甙为对照品,蛋糕中栀子黄经提取净化后,作为供试品,用高效液相色谱法测定,以保留时间定性,峰面积定量进行含量测定。     

Perchlorate transport and inhibition of the sodium iodide symporter measured with the yellow fluorescent protein variant YFP-H148Q/I152L

Perchlorate is an environmental contaminant that impairs thyroid function by interacting with the sodium iodide symporter (NIS), the transporter responsible for iodide uptake in the thyroid gland. Perchlorate is well known as a competitive inhibitor of iodide transport by NIS, and recent evidence demonstrates that NIS can also transport perchlorate. In this study, we evaluated the yellow fluorescent protein (YFP) variant YFP-H148Q/I152L, as a genetically encodable biosensor of intracellular perchlorate concentration monitored by real-time fluorescence microscopy. Fluorescence of recombinant YFP-H148Q/I152L was suppressed by perchlorate and iodide with similar affinities of 1.2 mM and 1.6 mM, respectively. Perchlorate suppressed YFP-H148Q/I152L fluorescence in FRTL-5 thyroid cells and NIS-expressing COS-7 cells, but had no effect on COS-7 cells lacking NIS. Fluorescence changes in FRTL-5 cells were Na⁺-dependent, consistent with the Na⁺-dependence of NIS activity. Perchlorate uptake in FRTL-5 cells resulted in 10-fold lower intracellular concentrations than iodide uptake, and was characterized by a higher affinity (K_m 4.6 μM for perchlorate and 34.8 μM for iodide) and lower maximal velocity (V_max 6.8 μM/s for perchlorate and 39.5 μM/s for iodide). Perchlorate also prevented iodide-induced changes in YFP-H148Q/I152L fluorescence in FRTL-5 cells, with half-maximal inhibition occurring at 1.1-1.6 μM. In conclusion, YFP-H148Q/I152L detects perchlorate accumulation by thyroid and other NIS-expressing cells, and reveals differences in the kinetics of perchlorate versus iodide transport by NIS.