青蒿素类药物专利技术分布研究

青蒿素是中国科学家自主创新的倍半萜内酯类抗疟化合物。经几十年的发展，该类药物已经成为
治疗疟疾的首选药物。然而，在巨大的国际市场中，中国企业却基本处于产业链的底端。该文力图从专利保
护的角度分析中国青蒿素产业缺乏市场竞争力的原因，并提出解决问题的建议。     

青蒿素的提取,分离和测定

应用薄层层析法对凤县不同产地青蒿中的青蒿素进行了测定。测定结果表明,凤县青蒿中青蒿素含量为０．１９６－０．２３０％。     

青蒿素及其衍生物的抗疟机制研究进展

青蒿素及其衍生物是一类重要的抗疟药物,其分子结构中含有特殊的过氧桥键.近几十年来,青蒿素及其衍生物的抗疟机制引起了国内外科学工作者浓厚的兴趣.文中结合最新的研究结果,综述了青蒿素及其衍生物的抗疟机制.研究进展表明,该类衍生物的抗疟机制基本分为两步：首先,青蒿素及其衍生物在疟原虫体内的Fe(Ⅱ)或其他还原性物质作用下形成一些活性中间体;然后,这些中间体通过过氧化膜脂质、干扰线粒体的功能、烷基化血红素或抑制蛋白质活性等途径表现出抗疟作用.作者对3种可能的活性中间体学说以及作用靶点进行了详细的讨论,并提出了下一步的研究方向.通过对青蒿素及其衍生物抗疟机制的论述,为今后抗疟药物的设计提供帮助.     

RH-HPLC法测定青蒿素中脱氢青蒿素及9-表-青蒿素的含量

建立了一种利用反相液相色谱法测定青蒿素原料药中脱氢青蒿素和9-表-青蒿素含量的分析方法。采用紫外检测器于210 nm处进行检测,以C-18柱(Agilent Zobax ODS,150 mm×4.6 mm,5μm)为分析柱,以乙腈和水的混合溶液为流动相,采取梯度洗脱方式(0~10 min,45%乙腈;10~20 min,45%~80%乙腈;20~25 min,80%~45%乙腈;25~35 min,45%乙腈)进行洗脱。在此条件下,青蒿素、脱氢青蒿素和9-表-青蒿素能彼此完全分离。脱氢青蒿素和9-表-青蒿素的浓度分别在4.11~28.76、9.42~65.93μg/m L范围内与峰面积的线性关系良好,相关系数R均大于0.998,检出限(3S/N)分别为2.668 ng和75.34 ng,样品加标回收率均在94%~108%之间。     

青蒿素传奇

疟疾,民间俗称“打摆子”,是经按蚊叮咬或输入带疟原虫的血液而感染的虫媒传染病。雌性按蚊是疟原虫的携带者和传播疟疾的罪魁祸首。根据寄生于人体的疟原虫种类,疟疾可分为间日疟、三日疟、恶性疟及卵围疟,主要表现为周期性规律发作。成人疟疾患者一般先全身发冷,并有明显的寒战,10分钟至24小时后体温迅速上升,持续高热2～6小时后,患者全身大汗淋漓,     

抗心律失常药物作用最佳靶点研究

目的:通过研究乌头碱、哇巴因致心律失常作用的离子靶点,揭示抗心律失常药物作用的最佳靶点。方法:采用全细胞膜片钳技术研究乌头碱、硅巴因对大鼠单个心室肌细胞动作电位时程(APD)、L-型钙电流(ICa)、钠电流(INa)、内向整流钾电流(IK1)以及瞬时外向钾电流(Ito)的作用。结果:乌头碱(1μM)使大鼠单个心室肌细胞90%复极化动作电位时程(APD90)从给药前的150.23±7.02ms延长至236.03±23.22ms(n=8,p<0.01)。ICa在0mV刺激电位下从-727.9±178.0pA增加至-1082.1±222.2pA(n=6,p<0.01);IK1在-110mV刺激电位下从-2122.0±511.1pA增加到-2536.3±386.5pA;乌头碱(1μM)抑制Ito,在+50mV刺激电压下,Ito从3203.6±617.1pA下降到2809.0±547.3pA。同时增加INa。哇巴因5μM使大鼠心肌细胞APD缩短(APD90从86.3±25.2ms缩短至58.9±20.8msn=5,p<0.01),ICa在+10mV刺激电位下从-1326.9±318.9pA减少到-782.3±395.3pA;使Ik1从-1868.3±187.8pA增加到-2392.8±366.7pA(刺激电压-120mV,n=10,p<0.01);使Ito从1272.7±317.6pA增加到1706.6±485.5pA(刺激电压+60mV,n=5,p<0.01)。结论:乌头碱、哇巴因诱发心律失常发生的活性点或最佳靶点是ICa,INa,IK1,Ikr和Iks。APD缩短或过度延长均可致心律失常,一个理想的抗心律失常药物应对心     

中药能遏制寄生虫抗药性

据《自然》杂志报道 ,经过十多年的研究 ,一种基于中国传统中药——青蒿素研制出的药物能有效遏制疟疾寄生虫的抗药性 ,他们认为这种新药也许能革命性地改变疟疾的治疗方法。　　世界上每年有 3亿人患疟疾 ,其中 1 0 0万人因此死亡。从 2 0世纪 60年代开始 ,氯喹成为治疗疟疾的特效药之一 ,但在非洲和亚洲 ,疟疾寄生虫恶性疟原虫对氯喹等药物越来越具有抗药性。　　科学家一直在研究青蒿素杀死疟原虫的机理。部分研究人员认为 ,青蒿素中有一种名为氧化氢桥的化学结构 ,它在被铁离子分解后会形成活跃的自由基 ,能够对一系列蛋白质及其它的生…     

青蒿素与辅酶A作用的研究

首次提出了青蒿素分子以疟原虫体内的生物活性物质辅酶A分子作为靶标的抗疟机制的新模型,并从生化原理和实验上对青蒿素类药通过辅酶A对疟原虫的致死作用给出了合理的解释,获得了预期的结果.     

Global landscape of protein complexes in the yeast Saccharomyces cerevisiae

Identification of protein-protein interactions often provides insight into protein function, and many cellular processes are performed by stable protein complexes. We used tandem affinity purification to process 4,562 different tagged proteins of the yeast Saccharomyces cerevisiae. Each preparation was analysed by both matrix-assisted laser desorption/ionization-time of flight mass spectrometry and liquid chromatography tandem mass spectrometry to increase coverage and accuracy. Machine learning was used to integrate the mass spectrometry scores and assign probabilities to the protein-protein interactions. Among 4,087 different proteins identified with high confidence by mass spectrometry from 2,357 successful purifications, our core data set (median precision of 0.69) comprises 7,123 protein-protein interactions involving 2,708 proteins. A Markov clustering algorithm organized these interactions into 547 protein complexes averaging 4.9 subunits per complex, about half of them absent from the MIPS database, as well as 429 additional interactions between pairs of complexes. The data (all of which are available online) will help future studies on individual proteins as well as functional genomics and systems biology.     

Disruption of the actin cytoskeleton in yeast capping protein mutants

Capping protein controls the addition of actin subunits to the barbed end of actin filaments and nucleates actin polymerization in vitro. Capping protein has been identified in all eukaryotic cells examined so far; it is a heterodimer with subunits of relative molecular masses 32,000-36,000 (alpha-subunit) and 28,000-32,000 (beta-subunit). In skeletal muscle, capping protein (CapZ) probably binds the barbed ends of actin filaments at the Z line. The in vivo role of this protein in non-muscle cells is not known. We report here the characterization of CAP2, the single gene encoding the beta-subunit of capping protein in Saccharomyces cerevisiae. Yeast cells in which the CAP2 gene was disrupted by an insertion or a deletion had an abnormal actin distribution, including the loss of actin cables. The mutant cells were round and large, with a heterogeneous size distribution, and, although viable, grew more slowly than congenic wild-type cells. Chitin, a cell wall component restricted to the mother-bud junction in wild-type budding yeast, was found on the entire mother cell surface in the mutants. The phenotype of CAP2 disruption resembled that of temperature-sensitive mutations in the yeast actin gene ACT1, indicating that capping protein regulates actin-filament distribution in vivo.     

利用毕赤酵母表达植物甜蛋白（brazzein）的初步研究

实验将含有Brazzein基因的pPIC9K穿梭质粒通过电导入巴斯德比赤酵母（Pichia pastori）GS115中，并从22个阳性克隆中筛选出His^＋Mut^s高拷贝转化子2个，经诱导表达，产物进行Tricine-SDS-PAGE电泳鉴定，目标蛋白的相对分子量与理论值一致，又经小试（5～10L罐）蛋白产量可达385mg/L。利用大孔树脂D152初步分离，得到有微甜味的产物。进一步纯化后获得了¹D-NMR图谱。     

A yeast mitochondrial outer membrane protein essential for protein import and cell viability

The gene encoding ISP42, an integral outermembrane protein located at the yeast mitochondrial protein import site was cloned, sequenced and modified. Yeast cells depleted of ISP42 accumulate uncleaved mitochondrial precursor proteins and then die. ISP42 is the first mitochondrial membrane protein shown to be indispensable for protein import and cell viability.     

应用酵母双杂交系统筛选与NAP1相互作用的蛋白质

用NAP1作为"诱饵"蛋白,通过酵母双杂交系统筛选人淋巴细胞cDNA文库,鉴定阳性克隆;再利用酵母双杂交和免疫共沉淀验证相互作用. 结果表明,通过酵母双杂交系统筛选人淋巴细胞cDNA文库,阳性克隆的鉴定,发现了与NAP1的相互作用蛋白质―蛋白酶体α亚基3(PSMA3). 免疫共沉淀(Co-IP)结果证实外源表达的NAP1蛋白和PSMA3蛋白在293T细胞中有特异性的相互作用. NAP1作为FDC分泌的一种多肽,与PSMA3有特异性的相互作用,这种相互作用如何实现对蛋白酶体降解靶蛋白的活性调节,其作用机理有待深入研究.