丝裂素活化蛋白激酶参与内皮素刺激的兔主动脉平滑肌细胞增生

内皮素（ｅｎｄｏｔｈｅｌｉｎ,ＥＴ）是已知的体内活性最强的缩血管物质,其缩血管作用由Ｇ蛋白偶联受体所介导。但ＥＴ强大的促血管平滑肌细胞（ＶＳＭＣ）增生效应的机理尚未完全阐明。本研究选用培养的兔胸主动脉ＶＳＭＣ,探讨丝裂素活化蛋白激酶（ＭＡＰＫ）在ＥＴ促细胞增生中的作用。结果表明：ＥＴ－１呈时间和浓度依赖性地促进细胞摄取￣３Ｈ－ＴｄＲ和激活ＭＡＰＫ,此作用可被蛋白激酶Ｃ（ｐｒｏｔｅｉｎｋｉｎａｓｅＣ,ＰＫＣ）抑制剂Ｓｔａｕｒｏｓｐｏｒｉｎｅ（ＳＴＰ）,Ｈ－７和ＥＴ＿Ａ受体拮抗剂ＢＱ１２３所抑制,但不被酪氨酸激酶抑制剂ＨｅｒｂｉｍｙｃｉｎＡ（Ｈｅｒｂ）所抑制,用ＰＫＣ激动剂ＰＭＡ（Ｐｈｏｒｂｏｌｍｙｒｉｓｔａｔｅａｃｅｔａｔｅ）预处理ＶＳＭＣ,使其ＰＫＣ活性下调,可显著减弱ＥＴ－１对ＭＡＰＫ的激活能力。本结果提示：（１）ＭＡＰＫ参与ＥＴ－１所致的ＶＳＭＣ增生;（２）ＥＴ－１促细胞增生与激活ＭＡＰＫ的作用是由ＥＴ＿Ａ受体和ＰＫＣ介导的。     

Protoplast culture and plant regeneration ofPinellia ternata

A study was undertaken to develop a protoplast regeneration system for pinellia. A yield of 19 29 x 10⁵ protoplasts/g F. W. could be obtained from cell suspension cultures incubated in a digestion enzyme solution with 2% cellulase Onzuka R-10, 10% pectinase (Sigma), 0.01% pectolyase Y23. K8P and modified MS media were used to culture protoplasts in: a) liquid, b) liquid-solid double layer, or c) agarose embedded protoplast culture. The former two were conducive to colony formation from protoplast-derived cells. The frequency of cell division was about 8% after 3 days in culture. Gradually adding fresh medium of lower osmotic pressure into the medium for protoplast culture favored cell division. Calli (1–2 mm in diameter) formed after 30–40 days in culture. The calli transferred onto medium supplemented with KT (0.5 mg 1^–1) and NAA (0.2 mg 1)^–1) could regenerate plants after 40–50 days. Of 47 plantlets transplanted into plots, 29 flowered and were fertile.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid - KT kinetin - CH casein hydrolysate     

小檗科鬼臼亚科植物的核型研究

本文首次报道了中华山荷叶与川八角莲的核型,分别为Ｋ（２ｎ）＝１２＝８ｍ（４ＳＡＴ）＋２ｓｔ＋２ｔ及Ｋ（２ｎ）＝１２＝４ｍ（２ＳＡＴ）十４ｓｍ＋２ｓｔ（２ＳＡＴ）＋２ｔ,核型类型均为ＺＡ型。本文报道的桃儿七及八角莲的核型与前人的结果有一定差异,前者为：Ｋ（２ｎ）＝１２＝６ｍ（４ＳＡＴ）＋２ｓｍ＋２ｓｔ＋２ｔ,２Ｂ型,后者为Ｋ（２ｎ）＝１２＝８ｍ（２ＳＡＴ）＋２ｓｔ（２ＳＡＴ）＋２ｔ,为２Ａ型。本文分析了小檗科鬼臼亚科４个属共７种植物的核型,结果是该类植物的核型极为相似,染色体数目均为２ｎ＝１２,由８条ｍ或ｓｍ,２条ｓｔ以及２条ｔ染色体组成。核型的相似性反映了这类植物的亲缘关系,这４个属的植物是一个自然类群。但随着系统发育,核型的不对称性有所增加,其中以山荷叶属最为对称,八角莲属居中,桃儿七属与足叶草属最不对称。笔者认为,核型上的高度相似是该类植物在系统发育上不发达,属内种类稀少,通常为寡种属或单种属的重要原因。     

Affinity chromatography—dependent selection （ACDS） of genomic DNA fragments bound specifically to bacterial synthesized Myc／Myn proteins

This paper describes an approach to seek for mouse c-Myc/Myn proteins-bound specific sequences among genomic DNA.cDNA fragment of myn gene was obtained through RT-PCR technique from RNA of NIH3T3 cells.DNA fragments encoding BR/HLH/LZ structure of Myc and Myn proteins were cloned in frame into pGEX-2T vector respectively.Fusion GST-Myc and GST-Myn synthesized in E.coli hosts showed affinity to CACGTG E-box DNA and subsequently interacted with genomic fragments prepared through whole-genome-PCR.A PCR-assisted procedure which combines protein-DNA interaction and affinity chromatography was designed to enrich Myc/Myn bound DNA.At least two genomic DNA fragments obtained exhibit specifical binding capacity to Myc/Myn complex but not to GST alone.Significance of the work and of the technique itself as well asidentification of the DNAs are discussed.     

流行性出血热循环免疫复合物组分分析

应用ＳＤＳ－聚丙烯酰胺凝胶电泳及免疫转印技术对流行性出血热患才血清中免疫合物组分进行了分析。流行性出血热循环免疫复合物经ＳＤＳ－ＰＡＧＥ分离,考马斯亮兰染色,显色主要有７条带,分子量分别为２３ｋＤ,５０ｋＤ,５２ｋＤ,６５ｋＤ,７２ｋＤ,８０ｋＤ及１００ｋＤ。采用该病毒特异性抗血清、单克隆抗体以及人免疫球蛋白、补体成分抗血清识别,在其特环免疫复合物中可检出特异性病毒抗在及相应的免疫球状蛋白和补体成     

基因工程链激酶(r—SK)纯化及其单克隆抗体制备和鉴定

采用Rotofor等电聚焦和分子筛技术纯化大肠杆菌表达的重组链激酶(r—SK),其纯度为97%。比活性1×10⁶IU／mg,回收率41％。分析所纯化的r—SK　N端氨基酸顺序证实其1—15个氨基酸顺序与SK基因的核苷酸顺序所示3联密码子一致。以纯化r—SK为抗原,通过杂交瘤技术构建了分泌抗r—SK单克隆抗体(McAb)杂交瘤细胞(4D11),该McAb属IgG1亚型．特异地作用于r-SK和C组口溶血性链球菌分泌的SK。用底物显色法测定该McAb可抑制SK对纤溶酶原的激活。Western blot法证实该McAb能识别分子量为16 250Da的r—SK的CNBr裂解片段。推测SK蛋白中第71残基至第237残基间的氨基酸顺序参与SK与纤溶酶原的结合。     

抗人PAI—1单克隆抗体制备,鉴定和应用

以重组PAI-(rPAI-1)为抗原,通过杂交瘤技术获得6株阳性杂交瘤细胞(AP1、AP2、AP3、AP4、AP5和AP6),并用SPA亲和层析纯化了抗PAI-1单克隆抗体(McAb)。所有McAb均能识别rPAI和天然PAI-1,腹水滴度均为10~6以上。6种McAb对PAI-1亲和常数在3.45×10~7—1.05×10~9mol/L之间。AP2、AP3 McAb能完全抑制PAI-1活性,AP4、AP和AP6只能部分抑制PAI-1活性,AP1则不抑制PAI-1活性。6种McAb中只有AP1、AP4和AP5能识别PAI-1/t-PA复合物。利用AP1、AP3、和AP4 McAb制备免疫亲和柱一步纯化HepG_2细胞分泌的PAI-1,纯度大于98％,回收率92％,纯化倍数51倍。利用抗PAI-1 McAb建立了夹心法ELISA,测定了人血浆PAI-1水平,正常人血浆PAI-1平均含量(±D)为24.7±7.75ng/ml。     

云南阿昌族的红细胞血型分布

调查了１０２名云南阿昌族的ＡＢＯ,ＮＮＳｓ,Ｒｈ和Ｐ系统的红细胞血型,结果表明,阿昌族的基因频率ｐ（０．３８７４）是迄今国内调查过人群中的最高值,Ｅ（０．２４５９）和ＣＤｅ（０．６９３６）基因或染色体频率较高,而Ｓ（０．０６８６）和Ｐ１（０．１０８９）频率较低;Ｍｓ（０．５９５０）连锁率高于Ｎｓ（０．３３５３）;未发现ＳＳ和Ｒｈ（－Ｄ）阴性表现型。     

Ca^2＋敏感性微电极在心肌胞浆Ca^2＋活度检测中的应用

改进的中性载体Ｃａ２＋敏感性微电极（Ｃａ－ＩＳＥ）在尖端直径为０．４—０．８μｍ时仍具有良好特性,可用于心肌胞浆Ｃａ２＋活度（αＣａｉ）的检测。测得豚鼠右心乳头肌、狗心室肌和浦肯野纤维静息时分别为０．１９±０．０１（ｎ＝２２）,０．２０±０．０２（ｎ＝１１）和０．４６±０．０７（ｎ＝１３）μｍｏｌ·Ｌ－１。毒毛旋花苷Ｇ３μｍｏｌ·Ｌ－１使静态及动态心肌分别增高０．１８±０．０２和６．６９±２．０９μｍｏｌ·Ｌ－１（ｎ＝２２）,并出现触发活动（ＴＡ）。１００μｍｏｌ·Ｌ－１蝙蝠葛碱可抑制毒毛旋花苷Ｇ引起的增高,同时使其ＴＡ消失。表明Ｃａ－ＩＳＥ技术可用于心肌组织ＴＡ与的同步检测。     

Distribution of zinc metallothionein I mRNA in rat brain using in situ hybridization

Metallothionein (MT) isoforms I and II were first identified and characterized in our laboratories in several regions of brain, in hippocampal neurons in primary culture, and in retinoblastoma and neuroblastoma cell lines. In this study, by having employed the MT-I cDNA as a probe, we sought to gain additional insight about the function of MT by discerning the regional distribution of its mRNA. Northern blot analyses of brain mRNA revealed that the administration of zinc enhanced dramatically MT-I mRNA (570 bp). The in situ hybridization study revealed that MT-I mRNA was located in several areas of brain, with the highest concentrations found in the cerebellum, hippocampus, and ventricles. The results of these studies are interpreted to suggest that zinc enhances the synthesis of MT mRNA and MT in turn may participate in zinc associated functions in neurons.Abbreviations MT-I Metallothionein I isoform - mRNA Messenger ribonucleic acid - ³⁵S dCTP ³⁵S Deoxycytidine triphosphate - ³²P dCTP ³²P Deoxycytidine triphosphate - icv Intracerebroventricularly - IP Intraperitoneally - PBS Paraformaldehyde phosphate buffered saline solution - Tris 2 amino-2-hydroxymethylpropane-1,3 diol - EDTA Ethylenediaminetetraacetic acid - cDNA Complimentary deoxyribonucleic acid - bp Base pair