人Sema4C重组腺病毒载体的构建及其在小鼠成肌细胞系C2C12中的表达

利用Ad5腺病毒载体系统构建人Sema4C基因重组腺病毒表达载体并在成肌细胞系C2C12中表达,并初步探讨Sema4C基因在成肌发育过程中的可能作用。利用脂质体介导重组腺病毒载体转染HEK293细胞,包装出完整的腺病毒;将重组腺病毒载体感染C2C12成肌细胞后,利用激光共聚焦显微镜观察发现12h即有绿色荧光表达,24h后绿色荧光蛋白表达最强;流式细胞仪检测病毒的感染效率几乎达100%。WB检测结果表明感染重组腺病毒载体组C2C12细胞Sema4C蛋白的表达量明显高于空载体对照组（P<0.01）。为了进一步观察Sema4C基因对C2C12细胞增殖分化的影响,流式细胞仪检测了病毒感染48h后C2C12细胞的增殖指数,并对感染后诱导分化的C2C12细胞的分化情况进行了观察。我们的结果首次表明,过表达外源性人Sema4C基因不仅能使C2C12细胞的G0/G1期比例增加,细胞的增殖指数下降,同时在分化培养条件下还能促进C2C12细胞肌管的形成。     

Membranolysis by the ninth component of human complement

The lytic property of isolated C9 was shown by using heat (48°C) to induce polymerization of C9, which was then exposed to egg lecithin vesicles containing carboxyfluorescein. In this environment, C9 penetrated the vesicles and released the marker. C5b-9 and C5b-8 also released carboxyfluorescein at 48°C, as did C5b-7 to a lesser extent. However, neither isolated C5b-6, C7, C8 nor albumin caused such release. At 30°C, the C9 remained in the monomeric form and could not lyse the vesicles.C9 polymers formed at 48°C were ring-shaped with an internal diameter of approximately 100 Å and an outer diameter of approximately 180 Å. These rings of C9 polymers strikingly resembled the ultrastructures formed by the membrane attack complex of complement, viewed from the top.Our results indicate that polymeric C9 causes the membranolysis of phospholipid vesicles and, hence, that C9 alone is a cytotoxic molecule.     

绵羊Myostatin基因启动子的功能分析

相比于Myostatin基因的作用机制而言, 对Myostatin基因转录和表达的调控机制了解很少. 为了更好地了解Myostatin基因5’调控区的结构和功能, 深入研究Myostatin基因的转录调控机制, 在最近的研究中克隆了绵羊Myostatin基因的启动子(Pro)序列(GenBank 登录号为AY918121). 进一步以EGFP为报告基因, 构建了各种长度的野生型MSTNPro^W-EGFP载体和E-box元件突变型MSTNPro^EM-EGFP载体, 通过转染C2C12小鼠骨骼肌成肌细胞系或绵羊成纤维细胞后, 对各种情况下EGFP的瞬时或稳定表达进行荧光定量分析而比较不同条件下绵羊Myostatin基因启动子的转录调控活性. 结果表明, 0.3~1.2 kb的绵羊Myostatin基因启动子都能不同程度地驱动EGFP在C2C12细胞中的转录和表达, 其中1.2 kb片段的转录调控活性最高. 然 而, 将绵羊1.2 kb MSTNPro^W-EGFP载体转染绵羊成纤维细胞后并未观察到EGFP的表达, 说明Myostatin基因表达的肌肉特异性源于启动子的转录特异性. 对稳定转染绵羊1.2 kb MSTNPro^W-EGFP载体的C2C12细胞进行荧光分析, 结果表明细胞生长密度的增加可以反馈性抑制Myostatin基因的转录和表达. 在C2C12分化状态下, 1.2 kb野生型绵羊Myostatin基因启动子的活性比成肌状态时显著升高, 而1.2 kb E(3+5+7)突变型Myostatin基因启动子的活性在C2C12分化前后并未表现出差别. 这一结果暗示肌肉调控因子MyoD可能是通过与E-box结合而引起Myostatin基因在C2C12分化和生长状态时转录和表达差异的一个原因.     

中国海产刚毛藻科(Cladophoraceae)八个新记录种

报道中国海产刚毛藻科(Cladophoraceae),沙生刚毛藻Cladophora arenaria,蝾螺刚毛藻C. conchopheria,暗色刚毛藻C. opaca,透明刚毛藻C. pellucida,微小刚毛藻C. pusilla,棉形刚毛藻C. rudolphiana,光毛刚毛藻C. sericea,美丽刚毛藻C. speciosa。     

On the Genus Ceterachopsis (J. Sm.) Ching

eterachopsis was proposed by the senior author as an independent genus of the family Aspleniaceae in 1940 with 2 species. Since then much study on its morphology, anatomy, gametophyte and palynology has been carried out by Nayar, Bir, Chandra & Nayar and Chang et al., and they are of the opinion that the genus like Ceterach, is a comparatively primitive element in the family Aspleniaceae. The queer zigzag patern of cutting of lanceolate fronds appears extraordinary in the family Aspleniaceae, and also suggests its antiquity in evolution. So far only 3 species (C. dalhousiae, C. paucivenosa and C. magnifica) are recognized, the former from W. Himalayas, also known from Africa, while the latter two from W. S. China (Yunnan). However, in the past forty years the Chinese botanists have discovered 2 more species as new in N. W. Yunnan, thus bringing the total known species of the genus up to 5. The present paper is a brief summary on the genus Ceterachopsis, which will be published in detail in the Flora Sinica vol. 4. Pteridologists are also divided in their views regarding the generic status of Ceterachopsis with 5 well-defined species in Yunnan and the East Himalayas. We prefer to mintain it as a genus separate from Asplenium on account of its distinct morphological features. It is to be hoped that more species may come to light in the mountainsin N. W. Yunnan through further exploration now under way.     

siRNA介导的MKK4表达抑制研究

采用脂质体转染方法,将3对(siRNA ID#64447、siRNA ID#64539和siRNA ID#186583)人工合成的MKK4基因特异的小分子干扰RNA(siRNA)分别导入小鼠成肌细胞C2C12中.实时荧光定量PCR结果表明,转染了siRNA ID#64447的细胞,其MKK4 mRNA的表达水平下调得最多,约为85%.试验获得了能够高效下调MKK4基因表达的siRNA.     

Activation of protein kinase C by a tumor-promoting phorbol ester in pancreatic islets

Rat pancreatic islet homogenates display protein kinase C activity. This phospholipid-dependent and calcium-sensitive enzyme is activated by diacylglycerol or the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). In the presence of TPA, the Ka for Ca2+ is close to 5 microM. TPA does not affect phosphoinositide turnover but stimulates [32P]- and [3H]choline-labelling of phosphatidylcholine in intact islets. Exogenous phospholipase C stimulates insulin release, in a sustained and glucose-independent fashion. The secretory response to phospholipase C persists in media deprived of CaCl2. It is proposed that protein kinase C participates in the coupling of stimulus recognition to insulin release evoked by TPA, phospholipase C and, possibly, those secretatogues causing phosphoinositide breakdown in pancreatic islets.     

武夷山和张家界自然保护区的一些虫生真菌 I. 虫草属

从武夷山和张家界自然保护区发现三虫草新种,武夷山虫草Cordyceps wuyishanensis, 张家界虫草Cordyceps zhangjiajiensis和拟茂兰虫草Cordyceps maolanoides。武夷山虫草和其近缘种的主要区别是,可孕部分柱状、非多年生、子囊孢子不断裂和间细胞长达6～10祄。张家界虫草与其近缘种相比较的鉴别特征为非木质化的较小子座、子囊壳表生和具有较长的(15～23祄)次生子囊孢子。拟茂兰虫草和近缘种茂兰虫草C.maolanoides的形态特征相近,其主要差别是前者的子座和子囊壳都小得多。报道了玫烟色拟青霉Paecilomyces fumosoroseus与蜣螂虫草Cordyceps geotrupis 有密切关系。研究标本保存于贵州大学真菌资源研究室(LFRGU)。     

山西轮藻属新植物

本文报道采自山西省的轮藻属植物2个新种,运城轮藻Chara yunchengensis,拟灰色轮藻C.pseudocanescens和3个中国新记录,阿尔泰轮藻C.altaica,豪威轮藻C.howeana,味美轮藻簇毛变种C.delicatula var.barbata     

草鱼Annexin A4基因克隆、表达特性及进化

草鱼(Ctenopharyngodon idellus)Annexin A4基因克隆于肝肾cDNA文库,该cDNA全长为1307bp,其中,5′非编码区为104bp,3′非编码区为237bp,开放阅读框为966bp,编码321个氨基酸。草鱼Annexin A4编码的蛋白质N端由15个氨基酸残基组成,C端具有4个重复序列,每个重复序列都有一个Ⅱ型钙结合位点,在第4个钙结合位点中包含一个"KGD"序列。Annexin A4在草鱼肝、肾、脾、心、鳃、肠中的表达量均较高,而肌肉和脑中不表达;PolyⅠ:C诱导后,其表达均有所下调。系统发育树显示,草鱼Annexin A4与斑马鱼(Danio rerio)的进化距离最近。适应性进化分析没有检测到正选择位点,表明Annexin A4非常保守,受到强烈的功能约束。     

Phosphocalyculin C as a pyrophosphate protoxin of calyculin C in the marine sponge Discodermia calyx

Calyculin C, a minor derivative of the calyculins, has an additional methyl group on C32 of calyculin A. A recent biosynthetic study of calyculins revealed that an end product of calyculin biosynthesis is the pyrophosphate form, phosphocalyculin A. However, the pyrophosphate counterpart derived from calyculin C had not been reported. We isolated phosphocalyculin C as a minor pyrophosphate derivative, by a detailed investigation of an extract from the sponge Discodermia calyx. The treatment of phosphocalyculin C with the D. calyx cell-free extract significantly enhanced its cytotoxicity, providing molecular evidence for its role as the protoxin of calyculin C.     

人类锌指蛋白ZNF382对肌生成的调控作用

人类基因组中有大量哺乳类特有的KRAB锌指基因,这些基因的功能大多尚不清楚,ZNF382就是其中一个.本文研究了人类锌指蛋白ZNF382对肌生成的调控作用.人类ZNF382基因在肌肉组织中高表达,ZNF382的mRNA水平在C2C12细胞的肌生成过程中上调,ZNF382调节肌肉特异性基因的表达,在NIH3T3细胞中,ZNF382与E-box蛋白E12和成肌调节因子MyoD共转增强肌肉肌酸激酶MCK启动子的活性;ZNF382的启动子预测分析也发现ZNF382的启动子上含有MyoD和血清反应因子SRF的结合位点,这些结果提示人类ZNF382蛋白在肌生成中起着重要的作用.     

FR3D: finding local and composite recurrent structural motifs in RNA 3D structures

New methods are described for finding recurrent three-dimensional (3D) motifs in RNA atomic-resolution structures. Recurrent RNA 3D motifs are sets of RNA nucleotides with similar spatial arrangements. They can be local or composite. Local motifs comprise nucleotides that occur in the same hairpin or internal loop. Composite motifs comprise nucleotides belonging to three or more different RNA strand segments or molecules. We use a base-centered approach to construct efficient, yet exhaustive search procedures using geometric, symbolic, or mixed representations of RNA structure that we implement in a suite of MATLAB programs, “Find RNA 3D” (FR3D). The first modules of FR3D preprocess structure files to classify base-pair and -stacking interactions. Each base is represented geometrically by the position of its glycosidic nitrogen in 3D space and by the rotation matrix that describes its orientation with respect to a common frame. Base-pairing and base-stacking interactions are calculated from the base geometries and are represented symbolically according to the Leontis/Westhof basepairing classification, extended to include base-stacking. These data are stored and used to organize motif searches. For geometric searches, the user supplies the 3D structure of a query motif which FR3D uses to find and score geometrically similar candidate motifs, without regard to the sequential position of their nucleotides in the RNA chain or the identity of their bases. To score and rank candidate motifs, FR3D calculates a geometric discrepancy by rigidly rotating candidates to align optimally with the query motif and then comparing the relative orientations of the corresponding bases in the query and candidate motifs. Given the growing size of the RNA structure database, it is impossible to explicitly compute the discrepancy for all conceivable candidate motifs, even for motifs with less than ten nucleotides. The screening algorithm that we describe finds all candidate motifs whose geometric discrepancy with respect to the query motif falls below a user-specified cutoff discrepancy. This technique can be applied to RMSD searches. Candidate motifs identified geometrically may be further screened symbolically to identify those that contain particular basepair types or base-stacking arrangements or that conform to sequence continuity or nucleotide identity constraints. Purely symbolic searches for motifs containing user-defined sequence, continuity and interaction constraints have also been implemented. We demonstrate that FR3D finds all occurrences, both local and composite and with nucleotide substitutions, of sarcin/ricin and kink-turn motifs in the 23S and 5S ribosomal RNA 3D structures of the H. marismortui 50S ribosomal subunit and assigns the lowest discrepancy scores to bona fide examples of these motifs. The search algorithms have been optimized for speed to allow users to search the non-redundant RNA 3D structure database on a personal computer in a matter of minutes.     

浙江桂及其相近种的亲缘关系及分类等级探讨

浙江桂、普陀桂和日本桂(天竺桂)是樟属植物中的三个相近种。六十多年以来,围绕着它们的亲缘关系及其分类等级,植物学界展开了反复的讨论,至今仍然存在着较大的分歧。在文献考证的基础上,笔者通过赴模式产地考察与采集鉴定,从比较形态学、扫描电镜观察、种子油脂成份的化学分析和地理分布区等方面,进一步研究了这三个种的异同,认为浙江桂是中国大陆的特有种;普陀桂应当归并入日本桂,作为一个变种处理;日本桂的原变种在中国仅见产于台湾省,而在大陆并无自然分布。     

武夷山和张家界自然保护区的一些虫生真菌 I. 虫草属

从武夷山和张家界自然保护区发现三虫草新种,武夷山虫草Cordyceps wuyishanensis, 张家界虫草Cordyceps zhangjiajiensis 和拟茂兰虫草Cordyceps maolanoides.武夷山虫草和其近缘种的主要区别是,可孕部分柱状、非多年生、子囊孢子不断裂和间细胞长达6～10μm.张家界虫草与其近缘种相比较的鉴别特征为非木质化的较小子座、子囊壳表生和具有较长的(15～23μm)次生子囊孢子.拟茂兰虫草和近缘种茂兰虫草C.maolanoides的形态特征相近,其主要差别是前者的子座和子囊壳都小得多.报道了玫烟色拟青霉Paecilomyces fumosoroseus与蜣螂虫草Cordyceps geotrupis 有密切关系.研究标本保存于贵州大学真菌资源研究室(LFRGU).     

海北高寒草甸土壤有机碳同位素组成及C3/C4碳源的变化

通过对高寒嵩草草甸土壤剖面不同深度(0～5cm,5～15cm,15～25cm,25～35cm,35～50cm,50～65cm)有机碳稳定性碳同位素的测定发现,土壤有机碳稳定性同位素(δ^13C)随土壤深度的增加而变大。表层土壤(0～5cm,定义为现代土壤)的δ^13C值最小,基本上接近现代植被的碳同位素特征。在土层5～10cm深度以下(粗略地定为古土壤),土壤有机碳稳定性同位素骤然上升,与表层土壤的同位素特征明显不同。考虑到影响土壤碳同位素的诸多因素,通过稳定性碳同位素的质量平衡模型计算,得出初步结果：来自C4(或CAM)植物的碳源随土壤深度的增加而增大。进一步推测,该地区植被可能经历由C4植物占优势的群落向C3植物占优势的群溶演化的过程。在这个过程中,大气碳同位素的变化和土壤有机质的形成过程(有机质淋溶过程)等也会引起土壤碳同位素的升高,因此质量平衡模型可能会过多地估算C4组分,而低估C3组分。