CtBP/BARS induces fission of Golgi membranes by acylating lysophosphatidic acid

Membrane fission is essential in intracellular transport. Acyl-coenzyme As (acyl-CoAs) are important in lipid remodelling and are required for fission of COPI-coated vesicles. Here we show that CtBP/BARS, a protein that functions in the dynamics of Golgi tubules, is an essential component of the fission machinery operating at Golgi tubular networks, including Golgi compartments involved in protein transport and sorting. CtBP/BARS-induced fission was preceded by the formation of constricted sites in Golgi tubules, whose extreme curvature is likely to involve local changes in the membrane lipid composition. We find that CtBP/BARS uses acyl-CoA to selectively catalyse the acylation of lysophosphatidic acid to phosphatidic acid both in pure lipidic systems and in Golgi membranes, and that this reaction is essential for fission. Our results indicate a key role for lipid metabolic pathways in membrane fission.     

An ARF-GEF acting at the Golgi and in selective endocytosis in polarized plant cells

Circumstantial evidence suggests that intracellular membrane trafficking pathways diversified independently in the plant kingdom, but documented examples are rare. ARF-GEFs (guanine-nucleotide exchange factors for ADP-ribosylation factor GTPases) are essential for vesicular trafficking in all eukaryotic kingdoms, but of the eight ARF-GEF families, only the ancestral BIG and GBF types are found in plants. Whereas fungal and animal GBF proteins perform conserved functions at the Golgi, the Arabidopsis thaliana GBF protein GNOM is thought to act in only the process of recycling from endosomes. We now show that the related Arabidopsis GBF protein GNOM-LIKE1 (GNL1) has an ancestral function at the Golgi but is also required for selective internalization from the plasma membrane in the presence of brefeldin A (BFA). We identified gnl1 mutants that accumulated biosynthetic and recycling endoplasmic reticulum markers in enlarged internal compartments. Notably, in the absence of functional GNL1, Golgi stacks were rendered sensitive to the selective ARF-GEF inhibitor BFA, which caused them to fuse with the endoplasmic reticulum. Furthermore, in BFA-treated gnl1 roots, the internalization of a polar plasma-membrane marker, the auxin efflux carrier PIN2, was selectively inhibited. Thus, GNL1 is a BFA-resistant GBF protein that functions with a BFA-sensitive ARF-GEF both at the Golgi and in selective endocytosis, but not in recycling from endosomes. We propose that the evolution of endocytic trafficking in plants was accompanied by neofunctionalization within the GBF family, whereas in other kingdoms it occurred independently by elaboration of additional ARF-GEF families.     

Genome-wide analysis of human kinases in clathrin- and caveolae/raft-mediated endocytosis

Endocytosis is a key cellular process, encompassing different entry routes and endocytic compartments. To what extent endocytosis is subjected to high-order regulation by the cellular signalling machinery remains unclear. Using high-throughput RNA interference and automated image analysis, we explored the function of human kinases in two principal types of endocytosis: clathrin- and caveolae/raft-mediated endocytosis. We monitored this through infection of vesicular stomatitis virus, simian virus 40 and transferrin trafficking, and also through cell proliferation and apoptosis assays. Here we show that a high number of kinases are involved in endocytosis, and that each endocytic route is regulated by a specific kinase subset. Notably, one group of kinases exerted opposite effects on the two endocytic routes, suggesting coordinate regulation. Our analysis demonstrates that signalling functions such as those controlling cell adhesion, growth and proliferation, are built into the machinery of endocytosis to a much higher degree than previously recognized.     

NF-L and NF-M are expressed in epithelial cells and coassemble with keratin or vimentin

Neurofilaments (NFs) and cytokeratins are both heteropolymers, which assemble into intermediate filaments (IFs) only when other proper IF subunit proteins are expressed simultaneously. To study the assembly property of NFs, we constructed two recombinant adenovirus which could express NF-L or NF-M, fused with green fluorescent protein (GFP) respectively. Then they were introduced into vero cells, and expressed fusion protein. Double labels of GFP fluorescence and immunofluorescence staining indicated that NF-L-GFP or GFP-NF-M not only coassembled with endogenous vimentins, but also coassembled with keratins into a cytoplasmic network of filaments.     

Small G proteins are expressed ubiquitously in lymphoid cells and do not correspond to Bcl-2

The bcl-2 gene is consistently associated with t(14; 18) chromosomal translocations observed in a large fraction of human B-cell lymphomas. The t(14; 18) translocation results in deregulated expression of the bcl-2 gene and synthesis of inappropriately high levels of the Bcl-2 protein. Gene transfer studies suggest a role for Bcl-2 in cell survival, growth enhancement and oncogenic transformation. To test the suggestion that GTP-binding by Bcl-2 may mediate its biological effects we characterized the GTP-binding proteins in lymphoid cells expressing Bcl-2. Expression of several small GTP-binding proteins was found to be ubiquitous and did not vary with levels of Bcl-2. By using immunological, electrophoretic and cell-fractionation techniques, we separated Bcl-2 from G proteins of small relative molecular mass (Mr) and showed that it is incapable of binding GTP. Our results show that small Mr G proteins are widely expressed in lymphoid cells and that Bcl-2 is not a novel member of this GTP-binding protein family.     

维生素E及其与维生素C,维生素B2联用对小鼠抗 …

研究了维生素Ｅ（ＶＥ）及其与维生素Ｃ（ＶＣ）,维生素Ｂ２（ＶＢ２）联用对小鼠机体抗氧化机能的影响。结果表明,对小鼠单独饲喂ＶＣ或ＶＥ,ＶＣ,ＶＢ２联合饲喂,均能显著提高小鼠心、肝组织超氧化歧化酶（ＳＯＤ）的活性作用;单独饲喂ＶＥ,ＶＣ或ＶＥ,ＶＣ,ＶＢ２联合饲喂,其肝脏丙二醛（ＭＤＡ）含量均低于对照组,差异非常显著,肝脏ＳＯＤ／ＭＤＡ和ＧＳＨ－Ｐｘ／ＭＤＡ比值升高;单独饲喂ＶＣ组ＧＳＨ－Ｐｘ／ＭＤ     

数据仓库动态设计与维护中与／或有向图方法

数据仓库（DW）可以抽象地看成基本关系上的一组具体的视图，是一个随时间不断推移变化的动态实体，当有新的查询出现时，可以将它化成具体的视图表示并加入到数据仓库中。将数据仓库表示成与／或有向图能够较好地描述数据仓库中各种查询（视图）之间的关系，这样数据仓库动态设计问题就可以模拟成状态空间搜索问题，相应的转换规则就可以定义成状态转换规则，通过状态空间搜索的转换和化简来实现数据仓库的动态设计。讨论了将视图加入到数据仓库中的动态设计方法，并通过一个实际的例子介绍了这种动态数据仓库的建立过程。     

高效液相色谱法测量蔬菜水果中稀禾定的残留

采用反相高效液相色谱法研究了水果蔬菜中稀禾定残留的检测方法。样品用不同溶剂提取后,经分离净化,采用紫外检测器进行定性定量分析。在最佳实验参数下,稀禾定浓度在1.0~80.0μg/mL范围内与色谱峰面积成良好的线性关系,回收率为86.8%~90.5%,检出限为0.1μg/mL,测定结果的标准偏差为0.64,变异系数为1.66%。     

父母年龄和先天性唇（腭）裂患儿之间的关系

研究表明唇腭裂疾病的发生和其父母高年龄有关,但父母年龄在唇腭裂病因中的作用报道较少，且观点不一。本文旨在分析父母生育年龄和先天性唇腭裂患儿之间的关系。通过采用Logistic回归分析研究就诊于我院的675例非综合征性唇腭裂患者，包括404例唇/腭裂(cleft lip with/without cleft palate, CL±P)，271例腭裂(isolated cleft palate CP)，并记录父母生育年龄。根据裂隙的严重程度分为两组：唇/腭裂（CL±P）及腭裂（CP）。单侧裂分为左侧裂及右侧裂。研究发现在唇/腭裂（CL±P）中，父母双方年龄都大时，后代患唇/腭裂（CL±P）的风险要比父母双方都年轻的高很多。在腭裂（CP）中，父母年龄大比父母年轻的，后代更易患腭裂，但是无统计学意义。同时分析父母年龄和左右侧裂之间的关系。发现父母年龄和左右侧裂统计学无意义。     

维生素E及其与维生素C,维生素B2联用对小鼠抗氧化体系机能的影响

研究了维生素 E(VE)及其与维生素 C(VC) ,维生素 B2 (VB2 )联用对小鼠机体抗氧化机能的影响。结果表明 ,对小鼠单独饲喂 VC或 VE,VC,VB2 联合饲喂 ,均能显著提高小鼠心、肝组织超氧化歧化酶 (SOD)的活性作用 ;单独饲喂 VE,VC或 VE,VC,VB2 联合饲喂 ,其肝脏丙二醛 (MDA)含量均低于对照组 ,差异非常显著 ,肝脏 SOD/ MDA和 GSH-Px/ MDA比值升高 ;单独饲喂 VC组 GSH-Px/ MDA比值升高尤为明显 ;饲喂VE或 VC,VE,VB2 联合饲喂时 ,小鼠肝组织中谷胱甘肽过氧化物酶 (GSH-Px)活力有增强趋势。这表明 3种维生素联合使用时 ,可产生协同抗氧化、调血脂作用 ,这种作用有利于预防动脉粥样硬化的发生与发展。     

SGK and 14-3-3 protein are involved in the serine/ threonine phosphorylation mechanism for TPO/MPL signal transduction

Thrombopioetin (TPO), the critical regulator of platelet production, acts by binding to its cell surface receptor, c-Mpl. Yeast two-hybrid screening was performed to isolate the proteins interacting with the cytoplasmic domain of c-Mpl. 48 positive clones were isolated from 5 × 10⁶ independent transformants. The results of sequence analysis demonstrate that they represent 13 different protein encoding sequences. Among them there are a partial coding sequence of serine/threonine protein kinase SGK (serum and glucocorticoid-inducible kinase) and 14-3-3 theta protein partial coding sequence. GST-pull-down assay and co-immunoprecipitation in mammal cells have confirmed the interaction between these two proteins and c-Mpl. By constructing a series of deleted c-Mpl cytoplasmic domain, the interaction region in c-Mpl cytoplasmic tail was localized in amino acids 523–554. At the same time, the directed interaction between SGK and 14-3-3 proteins also has been verified by yeast two-hybrid assay. The present note is the first time to report that two proteins act with c-Mpl at the same time and put forward that SGK and 14-3-3 protein may be involved in the serine/threonine phosphorylation mechanism for signal transduction.     

Chiari畸形或/和脊髓空洞病人的SEPs变化模式和临床意义

以南京大学医学院1997年7月至2003年3月间收治的70例Chiari畸形／脊髓空洞症患的体感诱发电位的变化，探讨体感诱发电位在该病早期的诊断价值，对70例患进行双侧上下肢正中神经和胫后神经体感诱发电位检测．70例患中胫后神经SEPs检查有24例(34．3％)异常，其中Chiari畸形组2例，脊髓空洞组7例，Chiari畸形伴脊髓空洞组15例,70例患中正中神经SEPs检查有15例(21．4％)异常，其中Chiari畸形组1例，脊髓空洞组4例，Chiari畸形伴脊髓空洞组10例，胫后神经SEPs检查较正中神经SEPs更为敏感．常规SEPs能反应深感觉障碍，提示疾病的病损程度，但对Chiari畸形／脊髓空洞的早期诊断帮助不大。     

中药复方制剂骨科敷药对尺桡骨干骨折患者骨代谢的影响

目的探讨中药复方制剂骨科敷药对尺桡骨干骨折患者骨代谢的影响.方法选择46例闭合性尺桡骨干骨折住院患者为研究对象,分为治疗组(23例)和对照组(23例),观察骨科敷药对尺桡骨干骨折后患者骨代谢的改善情况.结果骨科敷药治疗组能够有效缓解骨折患者的肿胀、疼痛症状,在治疗周期内能够显著提高患者血清骨钙素和碱性磷酸酶水平,降低D-二聚体水平(P0.05),但对血清钙水平的影响无统计学意义.结论骨科敷药可以通过调节骨折患者的骨代谢而发挥促进骨折愈合的作用,同时通过降低血清D-二聚体水平而发挥抗凝血的活性.     

铁素体珠光体球铁强度和延伸率的计算细观力学分析

本文基于计算细观力学方法,用有限元网格模拟球铁细观结构,将搞拉强度判据和延伸率判据引入轴对称弹塑性大应变有限元程序,计算了在球墨体积百分数为12.566％、球化良好、分布均匀条件下。不同细观结构球铁的抗拉强度和延伸率。结果表明:(1)铁素体含量是影响铁素体球光体球铁抗拉强度的主要因素,其关系可用线性或双线性公式表示:(2)当球铁中铁素体含量在50％以上时,延伸率随铁素体含量增加而迅速增大;(3)当铁素体含量在50％以下时,延伸率主要由球光体组织性能控制,并受到铁素体分布形态的较大影响,以铁素体呈破碎状分布为优;(4)在一定尺寸范围内,球墨直径增大,将使铁素体球铁抗拉强度和延伸率提高;对球光体球铁,则使延伸率略为下降(小于1％),而抗拉强度提高。     

脉冲激光亦或电子束辐照对SUS316 L奥氏体不锈钢中空位扩散的影响

利用激光-超高压电子显微镜系统在500℃对SUS316L奥氏体不锈钢进行了电子束辐照和脉冲激光-电子束双束同时辐照(以下简称:同时辐照),通过观察和分析辐照后自由晶界处无空洞区域以及元素偏析,以电子束辐照结果为标准,对比研究了同时辐照对空位扩散的影响.结果表明:同时辐照后的无空洞区域宽度为48±16 nm,小于电子辐照的71±27 nm;不论在偏析程度还是偏析宽度上,同时辐照条件下Cr和Ni的偏析都小于对应的电子束辐照;同时辐照下空位的扩散通量仅为电子束辐照的45.7%.通过分析得出,和电子辐照相比,同时辐照促进了空位与间隙原子的再结合,限制空位向尾闾扩散,进而造成流入尾闾的空位数量减少,极大地抑制了辐照偏析与肿胀.脉冲激光-电子束双束同时辐照可以为探索抑制肿胀方法提供新的思路.     

表面活性剂及大分子作为纳米粒子制备模板的研究新进展

分析了表面活性剂、大分子以及表面活性剂-大分子复合体系在纳米粒子制备中的应用.研究表明,表面活性剂形成的胶束结构、大分子或表面活性剂和大分子形成的有序体均可以作为软模板,在制备纳米粒子中发挥模板作用和稳定保护作用.     

线性代数法在极弱酸、碱和混合酸碱滴定分析中的应用

提出了能测定任何酸、碱溶液各组分浓度的通用计算方程,线性方程组的解直接给出了各组分浓度。     

A thermodynamic model of calculating mass action concentrations for structural units or ion couples in NaClO4-H2O and NaF-H2O binary solutions and NaClO4-NaF-H2O ternary solution

A thermodynamic model of calculating mass action concentrations for structural units or ion couples in NaClO₄-H₂O and NaF-H₂O binary solutions and NaClO₄-NaF-H₂O ternary strong electrolyte aqueous solutions was developed based on the ion and molecule coexistence theory (IMCT). A transformation coefficient was needed to compare the calculated mass action concentration and the reported activity, because they were usually obtained at different standard states and concentration units. The results show that transformation coefficients between the calculated mass action concentrations and the reported activities of the same components change in a very narrow range. The transformed mass action concentrations of structural units or ion couples in NaClO₄-H₂O and NaF-H₂O binary solutions agree well with the reported activities. The transformed mass action concentrations of structural units or ion couples in NaClO₄-NaF-H₂O ternary solution are also in good agreement with the reported activities in a total ionic strength range from 0.1 to 0.9 mol/kg H₂O by the 0.1 mol/kg step with different ionic strength fractions of 0, 0.2, 0.4, 0.5, 0.6, 0.8, and 1, respectively. The results indicate that the developed thermodynamic model can reveal the structural characteristics of binary and ternary strong electrolyte aqueous solutions, and the calculated mass action concentrations of structural units or ion couples also strictly follow the mass action law.     

氮、磷限制对球形棕囊藻不同形态细胞生长和多糖产量的影响

为探究营养盐胁迫对广西北部湾不同倍性球形棕囊藻(Phaeocystis globosa)细胞生长和多糖产量的影响,本研究采用单因子控制试验,设置4.0∶1.0、24.5∶1.0和64.0∶1.0 3个氮、磷限制比例,对球形棕囊藻的单倍体和二倍体细胞进行人工海水培养,比较二者的生长变化及各生长期内藻多糖含量和透明胞外聚合颗粒物(Transparent Exopolymer Particles, TEP)的释放差异。结果表明,氮、磷营养对二倍体细胞前期生长的促进作用强于单倍体细胞,二倍体细胞生长率显著高于单倍体细胞(P<0.05)。球形棕囊藻细胞分泌的多糖主要以溶解性胞外多糖(Soluble Exopolysaccharide, sEPS)为主,其次是胞内多糖(Intracellular Polysaccharide, IPS),固着性胞外多糖(Bound Exopolysaccharide, bEPS)占总糖比例最低。单倍体细胞和二倍体细胞的bEPS分泌趋势相反。氮、磷限制组的单倍体单细胞胞外多糖(Extracellular Polysaccharide, EPS)、IPS和TEP...     

生长后期喷施石灰水或KH2PO4对甘蔗糖分积累、产量和某些生理特性的影响

用桂糖１１号和台糖１３４作为供试甘蔗品种，在生长后期（１０月１１日）叶面喷施石灰水和ＫＨ２ＰＯ４。结果提高了甘蔗叶片的Ｍｇ２＋－ＡＴＰ酶、Ｃａ２＋－ＡＴＰ酶、ＮＡＤＰ－苹果酸酶和中性转化酶活性以及蔗糖和水分含量，促进了甘蔗蔗糖分提高而使蔗汁还原糖分降低。石灰水处理甘蔗品质提高较快，而ＫＨ２ＰＯ４处理甘蔗品质提高效应维持时间相对较长。两种处理都有一定的增产作用，其中石灰水处理的增产幅度较大，与对照的差异达到了统计显著水平。ＫＨ２ＰＯ４处理还明显提高了甘蔗叶片的过氧化物酶、多酚氧化酶活性、呼吸强度和叶绿素含量。