Association of dystrophin-related protein with dystrophin-associated proteins in mdx mouse muscle.

Dystrophin is associated with a complex of muscle membrane (sarcolemmal) glycoproteins that provide a linkage to the extracellular matrix protein, laminin. The absence of dystrophin leads to a dramatic reduction of the dystrophin-associated proteins (156DAG, 59DAP, 50DAG, 43DAG and 35DAG) in the sarcolemma of patients with Duchenne muscular dystrophy and mdx mice. Here we demonstrate that dystrophin-related protein (DRP, utrophin), an autosomal homologue of dystrophin, is associated with an identical or antigenically similar complex of sarcolemmal proteins and that DRP and the dystrophin/DRP-associated proteins colocalize to the neuromuscular junction in Duchenne muscular dystrophy and mdx muscle. The DRP and dystrophin/DRP-associated proteins are found throughout the sarcolemma in small-calibre skeletal muscles and cardiac muscle of adult mdx mice. Because these muscles show minimal pathological changes, our results could provide a basis for the upregulation of DRP as a potential therapeutic approach.     

Deficiency of the 50K dystrophin-associated glycoprotein in severe childhood autosomal recessive muscular dystrophy.

X-linked recessive Duchenne muscular dystrophy (DMD) is caused by the absence of dystrophin, a membrane cytoskeletal protein. Dystrophin is associated with a large oligomeric complex of sarcolemmal glycoprotein. The dystrophin-glycoprotein complex has been proposed to span the sarcolemma to provide a link between the subsarcolemmal cytoskeleton and the extracellular matrix component, laminin. In DMD, the absence of dystrophin leads to a large reduction in all of the dystrophin-associated protein. We have investigated the possibility that a deficiency of a dystrophin-associated protein could be the cause of severe childhood autosomal recessive muscular dystrophy (SCARMD) with a DMD-like phenotype. Here we report the specific deficiency of the 50K dystrophin-associated glycoprotein (M(r) 50,000) in sarcolemma of SCARMD patients. Therefore, the loss of this glycoprotein is a common denominator of the pathological process leading to muscle cell necrosis in two forms of muscular dystrophy, DMD and SCARMD.     

Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage

Many observations suggest the presence of transmembrane linkages between the cytoskeleton and the extracellular matrix. In fibroblasts both light and electron microscopic observations reveal a co-alignment between actin filaments at the cell surface and extracellular fibronectin. These associations are seen at sites of cell matrix interaction, frequently along stress fibres and sometimes where these bundles of microfilaments terminate at adhesion plaques (focal contacts). Non-morphological evidence also indicates a functional linkage between the cytoskeleton and extracellular matrix. Addition of fibronectin to transformed cells induces flattening of the cells and a reorganization of the actin cytoskeleton, with the concomitant appearance of arrays of stress fibres. Conversely, disruption of the actin cytoskeleton by treatment with cytochalasin B leads to release of fibronectin from the cell surface. As yet, there is no detailed knowledge of the molecules involved in this transmembrane linkage, although several proteins have been suggested as candidates in the chain of attachment between bundles of actin filaments and the cytoplasmic face of the plasma membrane: these include vinculin, alpha-actinin and talin, each one having been identified at regions where bundles of actin filaments interact with the plasma membrane and underlying cell-surface fibronectin. Recently, the cell-substrate attachment (CSAT) antigen has been identified as a plasma membrane receptor for fibronectin, raising the possibility that this glycoprotein complex may serve as a bridge between fibronectin and one or more of the underlying cytoskeletal components mentioned. Here we have investigated the interaction of the purified CSAT antigen with these cytoskeletal components, and we demonstrate an interaction specifically between the CSAT antigen and talin.     

基于DAG方法的我国股票市场的联动分析

根据我国股票市场的关键性政策,将2001年2月19日至2009年7月30日分为4个样本时期,通过有向非循环图（DAG）和动态因果检验分析我国5个主要股票指数的联动效应．实证结果表明上证A股市场是导致深证A股市场变化的短期和长期原因,在B股市场刚对国内投资者开放时,上证和深讧B股的联动效应特别显著．近年来香港股票市场与国内其它市场的联动性越来越显著．     

糖芥总甙对大鼠心肌和大脑Na~ ,K~ -ATP酶活力的影响

用测定无机磷含量法研究了糖芥总甙对ｗｉｓｔａｒ大鼠心肌和大脑Ｎａ＋,Ｋ＋－ＡＴＰ酶活力的影响。实验结果表明,糖芥总甙能明显抑制大鼠心肌大脑Ｎａ＋,Ｋ＋－ＡＴＰ酶活力,且呈量效关系。实验证实糖芥总甙的强心机制与西地兰类强心甙相同     

Detection of K+ and Cl-channels from calf cardiac sarcolemma in planar lipid bilayer membranes

The ionic currents underlying the cardiac action potential are believed to be much more complex than those in nerve. During the cardiac action potential, various membrane channels control the flow of K+, Na+, Ca2+ and Cl- across the sarcolemma of cardiac muscle cells. Thus, it has become increasingly clear that a detailed knowledge of the mechanisms that activate (or inactivate) heart channels is required to understand cardiac excitability. We report here the use of planar lipid bilayer techniques to detect and characterize K+ and Cl- channels in purified heart sarcolemma membrane vesicles. We have identified four different types of channel on the basis of their selectivity, conductance and gating kinetics. We present in some detail the properties of a K+ channel and a Cl- channel. We have tentatively identified the K+ channel with the ix type of current found in Purkinje, myocardial ventricular and atrial fibres. The chloride channel might be related to the transient chloride current found in Purkinje fibres.     

The Duchenne muscular dystrophy gene product is localized in sarcolemma of human skeletal muscle

Duchenne muscular dystrophy (DMD) and its milder form, Becker muscular dystrophy (BMD), are allelic X-linked muscle disorders in man. The gene responsible for the disease has been cloned from knowledge of its map location at band Xp21 on the short arm of the X chromosome. The product of the DMD gene, a protein of relative molecular mass 400,000 (Mr 400K) recently named dystrophin, has been reported to co-purify with triads of mouse and rabbit skeletal muscle when assayed using polyclonal antibodies raised against fusion proteins encoded by regions of mouse DMD complementary DNA. Here we show that antibodies directed against synthetic peptides and fusion proteins derived from the N-terminal region of human DMD cDNA strongly react with an antigen present in skeletal muscle sarcolemma on cryostat sections of normal human muscle biopsies. This immunoreactivity is reduced or absent in muscle fibres from DMD patients but appears normal in muscle fibres from patients with other myopathic diseases. The same antibodies specifically react with a 400K protein in sodium dodecyl sulphate (SDS) extracts of normal human muscle subjected to Western blot analysis. We conclude that the product of the DMD gene is associated with the sarcolemma rather than with the triads and speculate that it strengthens the sarcolemma by anchoring elements of the internal cytoskeleton to the surface membrane.     

Y-152和K-156在果蝇ADH催化中的作用

果蝇醇脱氢酶酪氨酸－１５２（Ｙ－１５２）和赖氨酸－１５６（Ｋ－１５６）处于同类脱氢酶的保守位点。经人工定点突变和酶动力学分析,前者的苯丙氨酸（Ｙ１５２Ｆ）、组氨酸（Ｙ１５２Ｈ）和谷氨酸突变体（Ｙ１５２Ｅ）及后者的异亮氨酸突变体（Ｋ１５６Ｉ）均丧失催化活性。而半胱氨酸－１５２（Ｙ１５２Ｃ）及精氨酸－１５６（Ｋ１５６Ｒ）突变体活性分别是对应野生型的０．２５％和２．２％。此外,Ｙ１５２Ｃ和Ｋ１５６Ｒ的Ｋ     

Natural inhibitor of transforming growth factor-beta protects against scarring in experimental kidney disease.

The central pathological feature of human kidney disease that leads to kidney failure is the accumulation of extracellular matrix in glomeruli. Overexpression of transforming growth factor-beta (TGF-beta) underlies the accumulation of pathological matrix in experimental glomerulonephritis. Administration of an antibody raised against TGF-beta to glomerulonephritic rats suppresses glomerular matrix production and prevents matrix accumulation in the injured glomeruli. One of the matrix components induced by TGF-beta, the proteoglycan decorin, can bind TGF-beta and neutralize its biological activity, so decorin may be a natural regulator of TGF-beta (refs 3, 4). We tested whether decorin could antagonize the action of TGF-beta in vivo using the experimental glomerulonephritis model. We report here that administration of decorin inhibits the increased production of extracellular matrix and attenuates manifestations of disease, confirming our hypothesis. On the basis of our results, decorin may eventually prove to be clinically useful in diseases associated with overproduction of TGF-beta.     

Effects of simulated microgravity on the cell growth and extracellular matrix of cultured chondrocytes

The effects of simulated microgravity on cell growth and extracellular matrix of chondrocytes have been examined by a light microscope and a scanning electron microscope using cultured chicken embryonic chondrocytes as the model. No notable difference of the cell density between the rotation group and the control group has been found. But during the same period, the growth spots of the rotation group were sparser than those of the control group. These results indicated that the lower level of cell development and differentiation happened in the rotation group. Observation by the scanning electron microscope showed that the extracellualr matrix decreased after rotating, and the fibres in the extracellular matrix were slighter and blurrier than those of the control group. It is concluded that the simulated microgravity can affect the secreting and assembly of the extracellular matrix. The possible mechanism for them is discussed. Li Xiaobing made the same contribution to this work as the first author     

Interactions between the nuclear matrix proteins and the 5'-flanking cis-acting elements of the human ε-globin gene

An erythroid-specific nuclear matrix protein (termed ε-NMPk) in K562 cells, which can specifically bind to the positive stage-specific regulatory element (ε-PRE Ⅱ , - 446- - 419 bp) upstream of the human ε-globin gene, has been identified by using gel mobility shift assay. Meanwhile, Southwestern blotting assay showed that the nuclear matrix protein ε-NMPk in K562, cells may be composed of two polypeptides ( ～ 40 ku). In addition, it is observed in the gel mobility shift assay that the nuclear matrix proteins from K562, HEL and Raji cells can bind to the silencer DNA ( - 392- - 177 bp) in the 5'-flanking sequence of human ε-globin gene respectively. However, the shift band K detected in K562 cells is different from shift band H/R in HEL and Raji cells, suggesting that a common nuclear matrix protein may exist in HEL and Raji cells. Results show that the nuclear matrix protein may play an important role in the regulation of the human ε-globin gene expression.     

关于"基质"概念的研究进展及应用

基质存在于细胞内和细胞外(细胞间).细胞内基质也叫细胞液,由细胞膜包被;细胞外基质是细胞间质的重要组成部分,也叫细胞外液.两者在形态学特征上均为无定形胶胨状,结构组成上有相似之处.     

Fibronectin receptor structures in the VLA family of heterodimers

Multiple cell surface proteins of relative molecular mass 115,000-155,000 (Mr 115K-155K) have been implicated as receptors mediating adhesion to extracellular matrix proteins. But the organization and relatedness of these peptides has remained unclear. In separate studies, the 'very late antigens' VLA-1 (Mr 210K/130K) and VLA-2 (Mr 160K/130K) were initially characterized as surface heterodimers appearing 2-4 weeks after in vitro stimulation of human T cells. Three more VLA heterodimers have since been discovered, which, like VLA-1 and VLA-2, are each composed of unique alpha-subunits in association with a common 130K beta subunit. This paper shows that the common VLA beta-subunit is equivalent to subunits found in structures with known fibronectin and laminin receptor activity, and that VLA-3 and VLA-5 are similar or identical to these previously defined receptors for adhesion molecules. Antibody blocking studies confirmed that at least some of the widely distributed VLA proteins of previously unknown function are involved in cell adhesion to fibronectin and laminin. We suggest that the VLA family of receptors may provide cells with multiple independent substrate adhesion capabilities.     

Interactions between the nuclear matrix proteins and the 5′-flankingcis-acting elements of the human ε-globin gene

An erythroid-specific nuclear matrix protein (termed ε-NMP_k) in K562 cells, which can specifically bind to the positive stage-specific regulatory element (ε-PRE II, - 446 - 419 bp) upstream of the human ε-globin gene, has been identified by using gel mobility shift assay. Meanwhile, Southwestern blotting assay showed that the nuclear matrix protein ε-NMP_k in K562, cells may be composed of two polypeptides (∼ 40 ku). In addition, it is observed in the gel mobility shift assay that the nuclear matrix proteins from K562, HEL and Raji cells can bind to the silencer DNA (- 392 - 177 bp) in the 5′-flanking sequence of human ε-globin gene respectively. However, the shift band K detected in K562 cells is different from shift band H/R in HEL and Raji cells, suggesting that a common nuclear matrix protein may exist in HEL and Raji cells. Results show that the nuclear matrix protein may play an important role in the regulation of the human ε-globin gene expression.     

确定空间连杆机构运动误差的微分算子矩阵法

根据矩阵闭环方程和误差分析理论,提出了计算空间连杆机构运动误差的微分算子矩阵法。灵活应用该文导出的微分算子矩阵和有关公式,可以得到简洁的矩阵形式的误差计算公式。该文方法特别适用于只含Ｒ、Ｐ、Ｃ、Ｈ运动副的空间连杆机构。更有意义的是所有公式的数值计算均可由计算机自动完成,从而为编制这类机构运动误差分析的通用程序奠定了理论基础     

基于灰度DAG熵最大化量化分辨率医学图像增强

为提高医学图像增强的清晰度和对比度,并提高计算效率,提出一种基于灰度有向无环图(DAG)熵最大化量化分辨率医学图像色调增强算法.首先,采用简单的分段自回归(PAR)模型进行图像目标恢复,并考虑到模数转换的误差利用全最小二乘算法进行PAR模型参数估计,获得高分辨率图像恢复直方图模型;其次,针对可能存在的对比度过低问题,将上述获得的最小二乘算法约束优化问题,建模为DAG中的最大权重路径问题,构建了色调保持最大熵图像增强过程约束优化模型,并通过DAG图Monge定理特性实现计算复杂度的降低;通过上述两个步骤,实现了医学图像增强过程中图像细节和对比度的同步增强,仿真实验显示所提算法可提供更为有效的医学图像增强效果.     

Ankyrin binding to (Na+ + K+)ATPase and implications for the organization of membrane domains in polarized cells

The interaction between membrane proteins and cytoplasmic structural proteins is thought to be one mechanism for maintaining the spatial order of proteins within functional domains on the plasma membrane. Such interactions have been characterized extensively in the human erythrocyte, where a dense, cytoplasmic matrix of proteins comprised mainly of spectrin and actin, is attached through a linker protein, ankyrin, to the anion transporter (Band 3). In several nonerythroid cell types, including neurons, exocrine cells and polarized epithelial cells homologues of ankyrin and spectrin (fodrin) are localized in specific membrane domains. Although these results suggest a functional linkage between ankyrin and fodrin and integral membrane proteins in the maintenance of membrane domains in nonerythroid cells, there has been little direct evidence of specific molecular interactions. Using a direct biological and chemical approach, we show here that ankyrin binds to the ubiquitous (Na+ + K+)ATPase, which has an asymmetrical distribution in polarized cells.     

Regional specialization of retinal glial cell membrane

Neural activity generates increases in extracellular K+ concentration, [K+]0, which must be regulated in order to maintain normal brain function. Glial cells are thought to play an important part in this regulation through the process of K+ spatial buffering: K+-mediated current flow through glial cells redistributes extracellular K+ following localized [K+]0 increases. As is the case in other glia, the retinal Müller cell is permeable almost exclusively to K+ . Recent experiments have suggested that this K+ conductance may not be distributed uniformly over the cell surface. In the present study, two novel techniques have been used to assess the Müller cell K+ conductance distribution. The results demonstrate that 94% of all membrane conductance lies in the endfoot process of the cell. This strikingly asymmetric distribution has important consequences for theories concerning K+ buffering and should help to explain the generation of the electroretinogram.     

Regulation of vertebrate left-right asymmetries by extracellular matrix.

The vertebrate body is organized along three geometric axes: anterior-posterior, dorsal-ventral and left-right. Left-right axis formation, displayed in heart and gut development, is the least understood, even though it has been studied for many years. In Xenopus laevis gastrulae, a fibronectin-rich extracellular matrix is deposited on the basal surface of ectoderm cells over which cardiac and visceral primordia move during development. Here I report experiments in which localized perturbation of a small patch of extracellular matrix by microsurgery was correlated with localized randomization of left-right asymmetries. Global perturbation of the extracellular matrix by microinjection of Arg-Gly-Asp peptides or heparinase into the blastocoel resulted in global randomization of left-right asymmetries. From these observations, I suggest that left-right axial information is contained in the extracellular matrix early in development and is independently transmitted to cardiac and visceral primordia.