Orthogonal solid-phase synthesis of a monobiotinylated analog of Neuropeptide Y

Analogs of Neuropeptide Y (NPY) were synthesized with conventional Boc/benzyl protective group strategy. Instead of Asn⁷ in the native scquence, Boc-Lys(Alloc)-OH was incorporated. At the end of the synthesis the Alloc group was selectively removed by palladium-catalyzed hydrostannolysis and biotin coupled to the e-amino group of Lys⁷. After cleavage and characterization with plasma desorption mass spectrometry the N^e-7-biotinyl-[Lys⁷]-NPY and the nonbiotinylated analog [Lys⁷]-NPY were investigated as ligands to the NPY receptor from rat cerebral cortex. Both analogs were found to be high affinity ligands to the NPY receptor and bound with essentially the same affinity as unmodified NPY.     

树型金银花无公害优质加工技术的研究

目的探讨树型金银花无公害优质加工技术。方法采用树型金银花晾晒和烘烤以及"烤房3段变温烘烤技术"。结果烘干产品比晒干产品具有明显的质量和价格优势,价格高57.1%。烤房3段变温技术第1段烘干温度控制在35～40℃,烘8～9h;第2段将温度提高到50～55℃左右,烘烤6～8h;第3段温度升到60～70℃,烘烤5～6h左右;烘干总时间不超过24h。结论无公害优质加工技术利于规模化生产,获得无公害的绿色产品,增加金银花附加值。     

X刀治疗脑内病变283例长期随访研究

目的:总结X刀治疗脑内病变的长期随访结果.方法:对283例X刀治疗的脑瘤和脑血管畸形患者进行临床和影像学随访,随访时间为1～8年,平均(2.05±4.66)年.结果:82例转移瘤消失35例(42.7％),缩小28例(34.1％),无变化11例(13.4％),增大8例(9.8％);61例胶质瘤消失10例(16.4％),缩小19例(31.1％),无变化16例(26.2％),增大16例(26.2％);55例动静脉畸形消失29例(52.7％),缩小20例(36.4％),无变化6例(10.9％);17例海绵状血管瘤缩小4例(23.5％),无变化13例(76.5％);35例脑膜瘤消失5例(14.3％),缩小15例(42.9％),无变化13例(37.1％),增大2例(5.7％);12例神经鞘瘤消失3例(25.0％),缩小6例(50.0％),无变化2例(16.7％),增大1例(8.3％);10例生殖细胞瘤消失8例(80.0％),缩小2例(20.0％);11例其他肿瘤消失2例(18.2％),缩小4例(36.4％),无变化4例(36.4％),增大1例(9.0％).影像检查还发现病灶有坏死表现99例,囊性变19例,周边环状强化73例,胶质细胞增生20例,暂时性反应性肿大23例,放射性脑水肿28例.结论:X刀对小型脑肿瘤和血管畸形是一种安全有效的治疗方法,X刀治疗的主要并发症是放射性脑水肿.     

反义核酸技术及应用

反义核酸技术是用人工合成的或生物合成的特定互补的DNA或RNA片段阻断从基因到蛋白的信息流 ,以达到抑制或阻断基因表达的生物新技术。利用核酸分子杂交原理将反义寡核苷酸 (antisenseoligonucleotide,ASON)作为外源性基因表达抑制物结合到靶mRNA序列上阻止其翻译成蛋白[1 ] 。具有杂交特异性和选择性调节基因的表达的特点[2 ] ,这一技术不仅已成为常用的实验室研究方法 ,还广泛的应用于研究病毒、肿瘤、寄生虫和遗传性疾病的预防和治疗 ,具有重要的应用前景。对于人类基因的研究 ,人们采用分子克隆、分子杂交、PCR技术等一系列研究…     

吸收度线性组合法测定盐酸乙基吗啡滴眼液的含量

本文应用吸收度线性组合法,不经分离直接测定盐酸乙基吗啡滴眼液的含量。以258、282、283及284nm波长为测试点,分别在各点测定盐酸乙基吗啡和尼泊金乙酯混合液的吸收度,通过各点的线性组合,计算盐酸乙基吗啡的含量。其回收率为100.12%.CV%=0.21%.本方法是测定盐酸乙基吗啡滴眼液的新方法,具有快速、准确的特点。     

Hemoglobin constant spring defined by specific oligonucleotide hybridization and hemoglobin D Punjab (beta 121----Gln) in a Batak Indonesian family

A Batak Indonesian from North Sumatra with hemoglobin (Hb) D Punjab (alpha 2 beta 2 121----Gln) and hemoglobin Constant Spring (Hb CoSp) is described. The 24-year-old man did not have clinical symptoms, and his hematological indices were normal. However, he had a persistent slight elevation of fetal hemoglobin level. His mother and his brother were heterozygous for Hb D Punjab; his father had Hb CoSp trait. A sister did not have any abnormal hemoglobin. To show the exact molecular defect leading to the synthesis of Hb CoSp in this family, genomic DNA from the father was analyzed by hybridization with synthetic oligonucleotides. Genomic DNA was digested with Sst I and Hind III producing a 1.05-kb fragment from the 3' end segment of the alpha 2-globin gene, including the termination codon. Two nonadecamers were synthesized to serve as probes: one, entirely homologous to the normal 3' end of alpha 2A-globin gene sequence, including the termination codon TAA, the other different from it by a replacement of the T in the termination codon TAA with C, changing it to CAA, the codon for the amino acid glutamine. DNA from normal controls gave a positive signal with the normal alpha 2TAA oligonucleotide probe but negative with the alpha 2 CAA probe. The father of propositus who had Hb CoSp trait gave a positive signal with the normal alpha 2TAA oligonucleotide probe as well as with the alpha 2CAA oligonucleotide probe, showing him to be heterozygous for the alpha 2CAA-globin gene. This result shows that the Hb CoSp in the Batak family is indeed due to a replacement of T by C in the TAA termination codon of the alpha 2-globin gene changing it to CAA the condon for glutamine. This explains the resulting readthrough of the untranslated sequence of the mRNA.     

核酸杂交反应中影响因素分析

以光敏生物素标记的R3 cDNA克隆为模型,采用正交设计的方法,对影响核酸杂交反应的主要因素:杂交反应温度,反应时间,探针浓度及甲酰胺浓度作了综合性分析。当探针浓度为800μg/L,甲酰胺浓度为10mol/L,50℃杂交反应6h,可获得最高灵敏度(10pg)。     

DNA探针的高效标记和哺乳动物细胞转化子的检测

本实验采用随机引物法标记探针DNA.探针DNA加热变性成单链,以单链DNA为模板,六聚脱氧核苷酸为引物,在DNA多聚酶I大片段的作用下进行放射性标记.标记探针的比放射性达10~8cpm/μg DNA以上.放射性底物的掺入率为60%.用该方法标记的人高度重复顺序探针,可以检出微微克水平的阳性分子.与人HeLa S_3细胞DNA转化的小鼠LTK~+细胞DNA打点杂交呈阳性,与未经转化的小鼠LTK~-细胞DNA杂交为阴性,证明小鼠转化细胞中存在HeLa细胞DNA,DNA转化是成功的.     

消渴降糖胶囊制备工艺的研究

目的 研究消渴降糖胶囊制备工艺.方法 采用高效液相色谱仪,以甲醇-0.4%磷酸溶液(60∶40)为流动相,检测波长为367 nm进行测定,记录色谱图,计算峰面积.结果 提取3 h、提取2 h与提取1 h相比,槲皮素提取率有显著增高(P<0.05),但提取3 h与提取2 h槲皮素的含量无显著性变化,浸膏得率3 h比2 h显著提高(P<0.05);加醇量4.0倍与3.0倍相比较,浸膏得率显著降低(P<0.05),槲皮素含量相差不大.结论 在制备消渴降糖胶囊工艺中,应减少能源消耗,降低无效成分的提取率,减少浸膏量.在尽量减少杂质,保留有效成分,降低服用量的综合考虑下,采用4.0倍量乙醇进行醇沉有效成分损失少,浸膏得率低,符合工业化要求.     

应用蛋白质组学技术在人肾癌组织中发现2个特异性生物标志物

目的探讨人肾肿瘤组织和正常肾组织之间蛋白质的差异表达，寻找特异性生物标记物。方法应用蛋白质组学（SELDI）技术，检测12例人肾肿瘤和正常肾组织蛋白质谱图表达。结果与正常肾组织比较，在IMAC3蛋白质芯片上人肾肿瘤组织谱图中显示2个特异性蛋白质峰明显增高，相对分子质量依次为5750.4和5861.7，其特异性和敏感性高。结论人肾肿瘤组织中特异性蛋白质的发现，对进一步研究肿瘤发生机理及其临床特异性生物标志物的检测等具有重要意义。