马氏珠母贝(Pinctada martenssi Dunker)外套膜组织培养基的设计

分析了培养基中的小牛血清、贝血清、水解乳蛋白等几种天然成分和合成培养基ＴＣ１９９对中国马氏珠母贝外套膜组织细胞在体外的生长情况和珍珠质分泌功能的影响，并确定了它们在组成培养基时的适宜含量．其中，贝组织提取物、鸡胚汁对细胞的生长和珍珠质的分泌具有十分重要的作用；５％的小牛血清和１０％的贝血清共同使用对维持细胞形促进细胞生长有很好的作用，但单独使用则有一定的负作用．     

Purification, Characterization and 1H NMR Resonance Assignment of an a－Like Neurotoxin BmK 16 from the Venom of Chinese Scor－pion Buthus martensii Karsch

A natural scorpion toxin BmK 16 was purified for the first time from the venom of the Chinese scorpion Buthus martensii Karsch (BmK) by using combined gel-filtration, ion exchange and reversed phase chromatography. The sequence of the N-terminal 8 amino acid residues was determined by Edman degradation. Using the N-terminal sequence as a tag, the database searching revealed a hit in the scorpion cDNA Bank. The sequence for N-terminal 8 amino acid residues, molecular weight and amino acid compositions of BmK 16 were identical with the calculated values according to the first 64 residues‘ se-quence of the precursor peptide alpha-neurotoxin TX16 derived from the sequence of the cDNA AF156597 (EMBL). The se-quence-specific resonance assignment of BmK 16 was achieved and the intact sequence of BmK 16 was determined as follow-ings: VRDAY IAKPH NCVYE CARNE YCNDL CTKNG AKSGY CQWVG KYGNG CWCKE LPDNV PIRVP GKCH. Furthermore, the results from the sequence homology analysis and the toxicity assays indicated that BmK 16 was an α-likescorpion neurotoxin.     

东亚钳蝎短链K+通道毒素BmP05基因多态性分析

基于从东亚钳蝎中鉴定的低电导Ca^2＋激活K^＋通道毒素BmP05的蛋白质和前体核苷酸序列，采用5’和3’RACE（rapid amplification of cDNA ends）技术，成功地从东亚钳蝎毒腺组织中分离到了一系列与BmP05前体核皆酸序列高同源性的cDNA序列，它们含有完整开放阅读框（open reading frame，ORF），彼此同源性很高，有的突变导致编码不同氨基酸，有的只足沉默突变，基因组克降结果进一步显示，尽管来源不同个体蝎的BmP05基因组的结构相同，但它们的基因组序列片不完全一致，在第二个外显子和内含子区域发生许多突变。BmP05在cDNA水平的多态性与其在基因组水平的多态性一致。所有结果表明，东亚钳蝎的毒素基因存在个体多态性。     

Structure and composition of the nacre–prisms transition in the shell of <Emphasis Type="Italic">Pinctada margaritifera</Emphasis> (Mollusca,Bivalvia)

A microstructural, mineralogical, and chemical study of the nacre–prisms boundary in the shells of Pinctada margaritifera shows that this boundary is not an abrupt transition, but that there exists a distinct fibrous layer with clear topographic structures and evidence of growth lines. A three-step biomineralization process is proposed that involves changes in the chemical and biochemical composition of the last growth increments of the calcite prisms, formation of the fibrous layer, and development of regular tablets in the nacreous layer.     

Purification and characterization of an analgesic peptide from Buthus martensii Karsch

The scorpion Buthus martensii Karsch (BmK) has been one of the indispensable materials in Chinese traditional medicine for thousands of years. In this work, an analgesic peptide was purified through four continuous chromatographic steps. The mouse twisting test was used to identify the target peptides in every separation step. The molecular weight, isoelectric point, and N-terminal residues of the purified peptide were determined. Based on the N-terminal sequence, the cDNA was also cloned by rapid amplification of cDNA ends from the cDNA pool of scorpion glands. This peptide was identical to BmK AS, an agonist of rabbit skeletal muscle ryanodine receptors. Preliminary pharmacodynamics revealed the following: the dose-effect curve plotted by the mouse twisting test showed an ED(50) of 1.42 mg/kg; and the time-effect curves plotted by a hot plate procedure showed a similar effect to the painkiller morphine. We report a purification procedure that yields substantial amounts of natural BmK AS having high activity. BmK AS has the potential to become a new analgesic medicine.     

Discovery of Guanidine Derivatives from Buthus martensii Karsch with Metal-Binding and Cholinesterase Inhibition Properties

Two rare guanidine-type alkaloids, Buthutin A (1) and Buthutin B (2), along with two other compounds (3, 4), were isolated from Buthus martensii Karsch, and determined using extensive spectroscopic data analysis and high resolution-mass spectrometry. Compound 1 showed the most potent inhibition on AChE and BChE with IC₅₀ values of 7.83 ± 0.06 and 47.44 ± 0.95 μM, respectively. Kinetic characterization of compound 1 confirmed a mixed-type of AChE inhibition mechanism in accordance with the docking results, which shows its interaction with both catalytic active (CAS) and peripheral anionic (PAS) sites. The specific binding of compound 1 to PAS domain of AChE was also confirmed experimentally. Moreover, compounds 1 and 3 exhibited satisfactory biometal binding abilities toward Cu²⁺, Fe²⁺, Zn²⁺ and Al³⁺ ions. These results provide a new evidence for further development and utilization of B. martensii in health and pharmaceutical products.     

马氏珠母贝遗传多样性的RAPD分析

采用 OPM组 2 0个碱基随机引物对海南三亚 (SY) ,海南洋浦港 (YP) ,广西涠洲岛 (WZ)的 3个天然群体和广东雷州人工养殖群体 (L Z)的马氏珠母贝进行了 RAPD分析 .其群体内的平均遗传相似度 (similarity,S)分别是 0 .5 86 6 ,0 .6 5 14,0 .6 6 6 6 ,0 .715 8,由低到高依次为 SYYP>WZ>L Z.结果表示 :天然群体的遗传多样性大于养殖群体的遗传多样性 .     

两个东亚钳蝎神经毒素cDNA的克隆和序列分析

构建了东亚钳蝎毒腺组织 c DNA文库 ,用 PCR筛选到两个新的神经毒素全长 c DNA,其开放阅读框(ORF)包括 :信号肽、成熟毒素和 /或额外氨基酸序列 ,它们编码的成熟毒素分别命名为 BmαTX9和 Bm IT( cp) 2 .序列分析表明 ,BmαTX9为抗哺乳动物神经毒素 ;Bm IT( cp) 2为痉挛型昆虫毒素 .文章还对东亚钳蝎神经毒素的命名 ,分类以及进化等问题进行了初步的探讨 .     

Purification, characterization and functional expression of a new peptide with an analgesic effect from Chinese scorpion Buthus martensii Karsch (BmK AGP-SYPU1)

In this study, a new peptide named BmK AGP‐SYPU1 with an analgesic effect was purified from the venom of Chinese scorpion Buthus martensi Karsch (BmK) through a four‐step chromatographic process. The mouse twisting test was used to identify the target peptides in every separation step. The purified BmK AGP‐SYPU1 was further qualified by RP‐HPLC and HPCE. The molecular mass determined by the MALDI‐4800‐TOF/TOF MS for BmK AGP‐SYPU1 was 7544 Da. Its primary structure of the N‐terminal was obtained using Edman degradation. The gene sequence of BmK AGP‐SYPU1 was cloned from the cDNA pool and genomic of scorpion glands, respectively, and then expressed in Escherichia coli. The sequence determination showed that BmK AGP‐SYPU1 was composed of 66 amino acid residues with a new primary structure. The metal chelating affinity column and cation exchange chromatography were used to purify the recombinant BmK AGP‐SYPU1. Consequently, the native and recombinant BmK AGP‐SYPU1 showed similar analgesic effects on mice as assayed using a mouse twisting model. These results suggested that BmK AGP‐SYPU1 is a new analgesic component found in the Chinese scorpion Buthus martensi Karsch. Copyright © 2010 John Wiley & Sons, Ltd.     

大珠母贝精子发生超微结构变化的研究

利用透射电子显微镜观察大珠母贝（ＰｉｎｃｔａｄａｍａｘｉｍａＪａｍｅｓｏｎ）的精子发生，揭示了从精原细胞逐渐发育成精子的超微结构的变化．精原细胞附着在基膜上，切面为扁平多边形；细胞核内开始形成板层小体．在初级精母细胞质中出现了前顶体粒（直径约０，２μｍ）和中心粒；核内板层小体的大小和数量在次级精母细胞时达到高峰．在精子细胞时期，一些核内板层小体进入细胞质中与前顶体粒融合形成顶体，一些保留在精核中，另一些则随残余细胞质被丢弃．精子结构为典型的原生型，由头部（长约２．０μｍ）、中段（长约０．８μｍ，宽约１．４μｍ）和尾部（长６５～７０μｍ，直径０．２μｍ）构成．顶体位于精子头部前端，呈中空的圆锥体．精核呈桶状；中段包括２个中心粒和４个紧密排列的线粒体；尾部鞭毛断面为典型的９＋２结构．