HBV─C基因探针的制备及应用

应用ＰＣＲ技术扩增出ＨＢＶＤＮＡＣ基因片段并与ｐＡＴ１５３质粒重组,转化到Ｅ．ｃｏｌｉＲＲＩ中,经体内扩增,提纯,用光生物素标记,制备了Ｃ基因的重组质粒探针。该探针检测灵敏度在Ｓｏｕｔｈｅｒｎ印迹中达１ｐｇ,在点印迹中为５ｐｇ。用此探针以Ｓｏｕｔｈｅｒｎ印迹方式配合ＰＣＲ技术检测乙肝病人血清中的ＨＢＶＤＮＡ,在５３例ＰＣＲ产物电泳检测阴性的样品中,Ｓｏｕｔｈｅｒｎ杂交又检出１８例阳性。     

PCR法快速检测临床标本中结核杆菌DNA

应用聚合酶链反应（ＰＣＲ）快速检测临床标本（脑脊液、胸水、腹水、血、痰液）中的结核杆菌ＤＮＡ,特异性扩增片段１２３ｂｐ,为结核杆菌的特异性重复序列ＩＳ６１１０部分基因。ＰＣＲ检测人型结核杆菌的敏感性达１０ｆｇＤＮＡ。临床标本的ＰＣＲ检测阳性率（２３．３％）明显高于抗酸染色涂片（２．９％）和细菌培养（５．７％）的阳性率（Ｐ〈０．０５）。通过设立对照系统及对扩增产物酶切分析,表明该法无假阴性结果（特异     

几种细胞分裂素诱导石竹外植体直接再分化成花芽的机理

用ＭＳ＋２,４－Ｄ０．１ｍｇ／Ｌ为基本培养基,分别附加不同浓度的ＺＴ、ＢＡ和ＫＴ,对石竹的叶片、茎和花等外植体直接再分化成花芽的试验表明：叶片的外植体在有ＢＡ时可再分化出只有红色花瓣的不正常花芽;在含ＺＴ的培养基中再分化出具有１～２片叶或无叶的花芽,其花有两性花,雌花和仅单个雌蕊的不正常花,两性花可发育成结籽的果实;ＫＴ未能诱导花芽分化。茎外植体,３种ＣＴＫ都不能诱导出花芽。花等外植体仅在ＫＴ３ｍｇ／Ｌ的培养中再分化出无叶的不正常花芽,ＺＴ和ＢＡ的处理均不能诱导出花芽。     

滇南及滇东南胶孔菌复合群的分类地理研究

报道了我国滇南及滇东南热带、亚热带胶孔菌复合群众路线属６种,其中瘤丝牛肝菌Ｆｉｌｏｂｏｌｅｔｕｓ ｖｅｒｒｕｃｕｌｏｓｕｓ Ｐ．Ｇ．Ｌｉｕ和滇丝牛肝菌Ｆ．ｙｕｎｎａｎｅｎｓｉｓ Ｐ．Ｇ．Ｌｉｕ是新种,紫兰小菇Ｍｙｃｅｎａ ｖｉｏｌａｃｅｌｌａ（Ｓｐｅｑ．）Ｓｉｎｇ。是我国新纪录种;新种附有拉丁文描术和插图,新纪录和附有形态解剖图。本文所引证标本均存放于中国科学院昆明植物研究所隐花植物标本馆（ＨＫＡ     

乌奴龙胆中五个新的环烯醚萜甙

从藏药乌奴龙胆（ＧｅｎｔｉａｎａｕｒｎｕｌａＳｍｉｔｈ）（龙胆科）的全草中分离到５个新的环烯醚萜甙,命名为乌奴龙胆甙（ｇｅｎｔｉｏｕｍｏｓｉｄｅ）Ａ－Ｅ;它们的结构主要通过光谱分析得以确定。其中,乌奴龙胆Ａ－Ｃ是二聚环烯醚萜甙,而乌奴龙胆甙Ｄ和Ｅ为马钱素型的环烯醚萜甙,所有这些化合物的分子中都具有一个２,３－二羟基苯甲酰基或其衍生物的取代基。     

中药玄参的化学成分

从中药玄参（ＳｃｒｏｐｈｕｌａｒｉａｎｉｎｇｐｏｅｎｓｉｓＨｅｍｓｌ．）中分离得到１个新化合物：４－Ｏ－（对甲氧基肉桂酰基）－α－Ｌ－鼠李糖［４－Ｏ－（ｐ－ｍｅｔｈｏｘｙｃｉｎｎａｍｏｙｌ）－α－Ｌ－ｒｈａｍｎｏｐｙｒａｎｏｓｅ］;同时还分离到４个已知的环烯醚萜甙：爪钧草甙（ｈａｒｐａｇｉｄｅ）,爪钩草酯甙（ｈａｒｐａｇｏｓｉｄｅ）,Ｏ－甲基梓醇（Ｏ－ｍｅｔｈｙｌｃａｔａｌｐｏｌ）,士可玄参甙Ａ（ｓｃｒｏｐｏｌｉｏｓｉｄｅＡ）和１个已知的苯丙素甙：安格洛甙Ｃ（ａｎｇｏｒｏｓｉｄｅＣ）。新化合物的结构通过化学降解和光谱分析得到证明。     

Colocalization of NADPH-diaphorase activity and certain neuropeptides in the esophagus of opossum (Didelphis virginiana)

Nitric oxide and various neuropeptides in the myenteric plexus regulate esophageal motility. We sought colocalization of nitric oxide synthase and neuropeptides in frozen sections of mid-portion of smoothmuscled opossum esophagus using NADPH-diaphorase activity to mark the synthase and immunoreactivity to detect peptides. The peptides, all with demonstrated physiological activity in this organ, were calcitonin generelated peptide, galanin, neuropeptide Y, substance P, and vasoactive intestinal polypeptide. The ExtrAvidin Peroxidase immunostain for each peptide was carried up to the final peroxidase reaction with 3-amino-9-ethylcarbazole. The NADPH-diaphorase reaction was applied with short incubation to provide light staining just before the peroxidase reaction was performed. We examined sections for the proportions of singly and dually labeled nerve cells in the myenteric plexus. NADPH-diaphorase activity was highly colocalized with calcitonin gene-related peptide (59%), galanin (54%), and vasoactive intestinal polypeptide (53%). It showed little colocalization with neuropeptide Y (10%) and substance P (8%). The proportions of all nerve cells containing each of the substances were: NADPH-diaphorase-33%, calcitonin gene-related peptide-30%, galanin-55%, neuropeptide Y-16%, substance P-35%, and vasoactive intestinal polypeptide-58%. We conclude that the nerves responsible for peristalsis in the esophagus may act by releasing nitric oxide along with other inhibitory substances, calcitonin gene-related peptide, galanin, and vasoactive intestinal polypeptide, but not excitatory substances, neuropeptide Y and substance P.     

日本血吸虫尾蚴发育的超微结构观察:Ⅰ.体被局部剖析

本文根据日本血吸虫尾蚴发育分期,除胚细胞（ＳＩ）已有报道（胡敏等,１９９１）外,对胚球期（Ｓ２）、尾蚴雏体期（Ｓ３）、成熟前期（Ｓ４）及成熟期（Ｓ５）在透射电镜对体被及其要素（ｔｅｇｕｍｅｎｔａｌ ｅｌｅｍｅｎｔｓ）进行剖析。结果原（始）体被最早出现于Ｓ２,表性分布。随后（Ｓ３—Ｓ５）消失而为真正体被所取代。体被皱褶亦随高隆。基膜及体棘为各期基本结构。感觉乳突于Ｓ３、Ｓ４出现。到了Ｓ５渐趋复杂而完善并对其结构作了较精细的描述。糖膜直至成熟期（Ｓ５）才出现,为尾期成熟的标志。近期对糖膜组份有较深入的研究并对其生理功能进行讨论。     

复合四倍体异育银鲫两种不同生殖方式的细胞学观察

在复合四倍体异育银鲫（）×银鲫（）的受精过程中,精子入卵后经过解凝、核化,最终形成雄性原核,并可与卵子的雌性原核融合,证明了复合四倍体异育银鲫卵子具有与两性融合生殖极为相似的拟两性融合生殖的能力;而在复合四倍体异育银鲫（）×兴国红鲤（）的组合中,精子入卵后以固缩状态存在,又表现出典型的雌核发育型生殖行为。因此我们认为复合四倍体异育银鲫具有两种不同的生殖发育机制。此外,我们还观察到在第一次有丝分裂中期有核物质被排斥到纺锤体之外的现象。本文就复合四倍体异育银鲫生殖发育的机制进行了初步的探讨。     

Agents that Promote Protein Phosphorylation Inhibit the Activity of the Na+/Ca2+ Exchanger and Prolong Ca2+ Transients in Bovine Chromaffin Cells

Abstract: The Na⁺/Ca²⁺ exchanger is an important element in the maintenance of calcium homeostasis in bovine chromaffin cells. The Na⁺/Ca²⁺ exchanger from other cell types has been extensively studied, but little is known about its regulation in the cell. We have investigated the role of reversible protein phosphorylation in the activity of the Na⁺/Ca²⁺ exchanger of these cells. Cells treated with 1 m M dibutyryl cyclic AMP (dbcAMP), 1 µ M phorbol 12,13-dibutyrate, 1 µ M okadaic acid, or 100 n M calyculin A showed lowered Na⁺/Ca²⁺ exchange activity and prolonged cytosolic Ca²⁺ transients caused by depolarization. A combination of 10 n M okadaic acid and 1 µ M dbcAMP synergistically inhibited Na⁺/Ca²⁺ exchange activity. Conversely, 50 µ M 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine, a protein kinase inhibitor, enhanced Na⁺/Ca²⁺ exchange activity. Moreover, we used cyclic AMP-dependent protein kinase and calcium phospholipid-dependent protein kinase catalytic subunits to phosphorylate isolated membrane vesicles and found that the Na⁺/Ca²⁺ exchange activity was inhibited by this treatment. These results indicate that reversible protein phosphorylation modulates the activity of the Na⁺/Ca²⁺ exchanger and suggest that modulation of the exchanger may play a role in the regulation of secretion.