Formation of domain structure of erythrocyte membrane in Wistar rat fed with CeCl3 per os

To explore the possibility of absorption of lanthanides via digestive duct and their effects on the membrane structure and permeability of erythrocytes, the fine structure of erythrocyte membrane from Wistar rats, fed for 70 days of daily administrationper os with 20 mg CeCl₃/kg weight, was imaged by means of atomic force microscopy and FT-IR deconvolution spectra. The results show that, although the erythrocytes maintain the intact shape, the change of secondary structure, aggregation and crosslinking of the protein particles of membrane surface and the enlarged lipid regions lead to the domain structure formation. This structure might be responsible for the increasing permeability of erythrocyte membrane.     

The asymmetric membrane structure of erythrocytes from Crucian carp studied by atomic force microscopy

Cell membranes play a key role in cellular activities. Fish erythrocytes, prototype of the nucleated erythrocytes of lower vertebrates, are predominantly oval, biconvex discs with elliptical nucleus and much larger in size than human erythrocytes. In attempts to disclose the correlation of membrane structure between fish and human erythrocytes, we used in situ atomic force microscope （AFM） combined with single-molecule force spectroscopy to study the membrane structure of Crucian carp erythrocytes under quasi-native conditions. Our results revealed the asymmetric distribution of proteins in Crucian carp erythrocyte membrane： the outer leaflet of membrane is rather smooth without any proteins, whereas the inner leaflet of membrane is very rough with dense proteins. The asymmetry of fish erythrocyte membrane structure fits well with the semi-mosaic model of human erythrocyte membrane structure. This similarity of membrane structure between human and fish erythrocytes extends the semimosaic model of erythrocytes membrane structure to a wider range of species.     

Partial deficiency of erythrocyte spectrin in hereditary spherocytosis

Hereditary spherocytosis (HS) is a common, clinically heterogeneous haemolytic anaemia in which the primary erythrocyte defect is believed to be some abnormality in the spectrin-actin membrane skeleton, leading to loss of surface membrane. Recessively inherited spectrin deficiency with extreme erythrocyte fragility and spherocytosis has been identified in certain mutant mice and two severely anaemic humans. Although suspected, deficiency of spectrin has not been demonstrated in less severe forms of human HS. We not report the quantitation of erythrocytes spectrin by radioimmunoassay. We found that normal erythrocytes contained 240,000 copies of spectrin heterodimer, whereas erythrocytes from 14 patients with a variety of types of HS were all partially deficient in spectrin (range 74,000-200,000 copies), the magnitude of the deficiency correlating with the severity of the disease. Spectrin deficiency of varying degrees is common in HS and probably represents the principal structural defect leading to loss of surface membrane.     

Abnormal display of PfEMP-1 on erythrocytes carrying haemoglobin C may protect against malaria

Haemoglobin C, which carries a glutamate-to-lysine mutation in the beta-globin chain, protects West African children against Plasmodium falciparum malaria. Mechanisms of protection are not established for the heterozygous (haemoglobin AC) or homozygous (haemoglobin CC) states. Here we report a marked effect of haemoglobin C on the cell-surface properties of P. falciparum-infected erythrocytes involved in pathogenesis. Relative to parasite-infected normal erythrocytes (haemoglobin AA), parasitized AC and CC erythrocytes show reduced adhesion to endothelial monolayers expressing CD36 and intercellular adhesion molecule-1 (ICAM-1). They also show impaired rosetting interactions with non-parasitized erythrocytes, and reduced agglutination in the presence of pooled sera from malaria-immune adults. Abnormal cell-surface display of the main variable cytoadherence ligand, PfEMP-1 (P. falciparum erythrocyte membrane protein-1), correlates with these findings. The abnormalities in PfEMP-1 display are associated with markers of erythrocyte senescence, and are greater in CC than in AC erythrocytes. Haemoglobin C might protect against malaria by reducing PfEMP-1-mediated adherence of parasitized erythrocytes, thereby mitigating the effects of their sequestration in the microvasculature.     

鲤鱼竖鳞病的血液病理学研究

从红细胞的形态，脆性和白细胞的比率以及血清电解质离子浓度和血清谷丙转氨酶等多方面对鲤鱼竖鳞病的血液病理改变进行了观察和研究。发现鲤鱼竖鳞病病鱼的红细胞膜出现皱缩，红细胞脆性增加，嗜中性粒细胞增多，血清谷丙转氨酶活性增高，血清中Ｎａ＾＝和Ｃａ＾２＋浓度极度降低，出现水肿和鳞片竖起。     

Reversible shape change of Triton-treated erythrocyte ghosts induced by Ca2+ and Mg-ATP

The shape of the human erythrocyte depends on intracellular ATP content and echinocytic erythrocyte ghosts obtained in the presence of Mg-ATP acquire a diskocytic shape after incubation at 37 degrees C; however, agreement is lacking about the molecular basis of the shape changes. The suggestion that phosphorylation of cytoskeletal structures underlying the membrane is involved has been disputed and alternative explanations based on lipid bilayer theory and metabolism of phospholipid have been proposed. Recently, we re-examined the effect of ATP on the shape of ghosts and found that it consists of two distinct steps. We have, therefore, now examined the effect of physiological concentrations of Ca2+ and ATP on the cytoskeleton of the erythrocyte membrane directly using Triton-treated ghosts which had lost this permeability barrier. Our findings suggest that a noncovalent effect of ATP on the cytoskeleton is a prerequisite for shape change.     

Structural basis for Duffy recognition by the malaria parasite Duffy-binding-like domain

Molecular processes that govern pathogenic features of erythrocyte invasion and cytoadherence in malaria are reliant on Plasmodium-specific Duffy-binding-like domains (DBLs). These cysteine-rich modules recognize diverse host cell-surface receptors during pathogenesis. DBLs of parasite erythrocyte-binding proteins mediate invasion, and those from the antigenically variant P. falciparum erythrocyte membrane protein 1 (PfEMP1) have been implicated in cytoadherence. The simian and human malarial parasites, P. knowlesi and P. vivax, invade human erythrocytes exclusively through the host DARC receptor (Duffy antigen receptor for chemokines). Here we present the crystal structure of the P. knowlesi DBL domain (Pkalpha-DBL), which binds to DARC during invasion of human erythrocytes. Pkalpha-DBL retains the overall fold observed in DBLs from P. falciparum erythrocyte-binding antigen (EBA)-175 (ref. 4). Mapping the residues that have previously been implicated in binding highlights a fairly flat but exposed site for DARC recognition in subdomain 2 of Pkalpha-DBL; this is in sharp contrast to receptor recognition by EBA-175 (ref. 4). In Pkalpha-DBL, the residues that contact DARC and the clusters of residues under immune pressure map to opposite surfaces of the DBL, and suggest a possible mechanism for immune evasion by P. vivax. Our comparative structural analysis of Pkalpha-DBL and P. falciparum EBA-175 provides a framework for the understanding of malaria parasite DBLs, and may affect the development of new prophylactic and therapeutic strategies.     

Modulation of spectrin-actin assembly by erythrocyte adducin

The spectrin-based membrane skeleton, an assembly of proteins tightly associated with the plasma membrane, determines the shape and mechanical properties of erythrocytes. Spectrin, the most abundant component of this assembly, is an elongated and flexible molecule that, with potentiation by protein 4.1, is cross-linked at its ends by short actin filaments to form a lattice beneath the membrane. These and other proteins stabilize the plasma membrane, organize integral membrane proteins and maintain specialized regions of the cell surface. A membrane-skeleton-associated calmodulin-binding protein of erythrocytes is a major substrate for Ca2+- and phospholipid-dependent protein kinase C (ref. 5), and thus is a target for Ca2+ by two regulatory pathways. Here we demonstrate that this protein, called adducin: (1) binds tightly in vitro to spectrin-actin complexes but with much less affinity either to spectrin or to actin alone; (2) promotes assembly of additional spectrin molecules onto actin filaments; and (3) is inhibited in its ability to induce the binding of additional spectrin molecules to actin by micromolar concentrations of calmodulin and Ca2+. Adducin may be involved in the action of Ca2+ on erythrocyte membrane skeleton and in the assembly of spectrin-actin complexes.     

红细胞形态特征的计算机图像处理研究

比较传统人工细胞形态观察和定量的操作方式，介绍了基于计算机图像处理和分析技术的有关红细胞形态特征定量的研究成果．并且展现了一套适用范围广、可作为红细胞图像的形态描述或细胞分类的图形特征参数——圆周率．同时，基于圆周率提供了一些较为适用的细胞形态特征值的算法(比如，多定位的图像获取、红细胞胞体形态的自动识别，以及对其进行的分类算法)．红细胞形态的算法或程序都基于VC ，不过该程序也可运用于其他用途．红细胞的染色采用瑞氏法(亚甲基蓝咿红)．介绍的红细胞形态计算机处理方法可提供有效的半自动红细胞形态特征的统计分析，可描述出群体细胞形态特征的分布曲线或不同细胞形状比率的数据．     

硒葡聚精对小鼠红细胞膜的保护作用

以外源性Ｈ２Ｏ２和羟自由基（·ＯＨ）为氧化诱导剂，观察了硒葡聚糖（Ｓｅｌｅｎｉｕｍ—ＧｌｕｃｏｓａｎＳｅ—Ｇｎ）对小鼠红细胞膜氧化损伤的保护作用。结果表明，硒葡聚糖可明显降低Ｈ２Ｏ２诱导的氧化溶血作用，下降率为１１％～２６％。红细胞内脂质过氧化物含量在Ｓｅ—Ｇｎ作用下明显降低。红细胞膜的荧光偏振度（Ｐ）也呈下降趋势，表明红细胞膜流动性增加。实验表明Ｓｅ—Ｇｎ对红细胞的保护作用可能与其参与抗氧化过程有关。     

红细胞沉降速率Ⅲ—红细胞可变形性的实验研究

介绍了一种利用红细胞沉降过程测量评价红细胞可变形性的新方法，并用它考查了猪红细胞在生理盐水溶液及ＳＦ６０溶液中的可变形性，实验得到的可变形性参数分别为０．８６２和０．９７１。     

Effect on membrane transport in the erythrocytes by band 3 cross-linking

Band 3 and glucose transport protein (GluT1) are two kinds of important proteins in the human erythrocyte membranes. Bis(sulfosuccinimidyl)suberate (BS³), an impermeable cross-linker of band 3, inhibited NO₂ ⁻ transport, showing that anion exchange is affected by the association state of band 3 in the intact erythrocyte membranes. At the same time, the rates of glucose transport of both exit and entry declined. The amount of monomers of band 3 was decreased after treatment of the erythrocytes with BS³, but there was no change in GluT1 according to the SDS-PAGE patterns. This demonstrates that band 3 and GluT1 would be linkaged together in the erythrocyte membranes for the requirement of rapid and cooperative performance of physiological functions of the membrane proteins.     

过度训练对红细胞膜结构和功能影响的实验研究

为进一步探讨过度训练的发生机制,采用建立一般训练和过度训练动物模型,应用荧光法和比色法分别测定了过度训练后红细胞膜MDA含量、红细胞变形性、Na~+-K~+-ATPase、Ca~(2+)-ATPase活性以及红细胞膜总磷脂、胆固醇含量及其比值.结果显示:与对照组相比,MDA含量在运动后即刻显著升高(P<0.01);红细胞变形性显著下降(<0.05;P<0.01),Na~+-K~+-ATPase和Ca~(2+)-ATPase活性显著下降(P<0.05),红细胞膜总磷脂显著下降(P<0.05),红细胞膜上胆固醇在运动后即刻显著低于对照组(P<0.01),红细胞膜上胆固醇/磷脂比值在运动后即刻显著低于对照组(P<0.05),相关分析表明,红细胞的变形性与红细胞膜上MDA含量呈显著负相关(P<0.05).结果揭示:过度训练运动引发的自由基产生的脂质过氧化对红细胞膜结构有损害,使Na~+-K~+-ATPase和Ca~(2+)-ATPase活性下降,红细胞膜上的磷脂和胆固醇含量以及其比值降低,致使红细胞变形性下降,可能是造成过度训练后红细胞功能下降的重要原因之一.     

Direct access to serum macromolecules by intraerythrocytic malaria parasites.

Trafficking pathways in malaria-infected erythrocytes are complex because the internal parasite is separated from the serum by the erythrocyte and parasitophorous vacuolar membranes. Intraerythrocytic Plasmodium falciparum parasites can endocytose dextrans, protein A and an IgG2a antibody. Here we show that these macromolecules do not cross the erythrocyte or parasitophorous vacuolar membranes, but rather gain direct access to the aqueous space surrounding the parasite through a parasitophorous duct. Evidence for this structure includes visualization of membranes that are continuous between the parasitophorous vacuolar and erythrocyte membranes, and surface labelling of the parasite with fluorescent macromolecules under conditions that block endocytosis. The parasite can internalize by fluid-phase endocytosis macromolecules from the aqueous compartment surrounding it. Thus, surface antigens on trophozoites and schizonts should be considered as targets for antibody-directed parasiticidal agents.     

胞二磷胆碱抑制贮存红细胞溶血作用的研究

贮存红细胞溶血的主要原因,现认为与能量代谢障碍和膜结构功能有密切关系.前文曾报导胞二磷胆碱(简称CDP胆碱)能抑制贮存红细胞的溶血,而且其作用主要不是通过能量代谢.本文试从CDP胆碱与红细胞膜磷脂的关系等方面作一些探索. 将去除血浆的红细胞贮存于代血浆中,看到CDP胆碱仍能抑制溶血.经测定血浆磷脂的含量,也没有发现CDP胆碱血和对照之间有明显差别.此结果提示CDP胆碱抑制红细胞溶血的作用与血浆并无联系,排除了通过与血浆进行磷脂交换来更新红细胞磷脂,从而抑制溶血的可能性.此外,还测定了CDP胆碱血和对照的红细胞膜磷脂含量,并用薄板层析法测定了卵磷脂和神经鞘磷脂的百分含量.结果显示,实验组并不比对照有所增高,这也表明CDP胆碱对红细胞膜磷脂含量没有显著增进作用. 除CDP胆碱以外,CMP也具有抑制贮存红细胞溶血的作用,其意义值得作进一步研究.     

胞二磷胆碱抑制贮存红细胞溶血作用的研究

贮存红细胞溶血的主要原因,现认为与能量代谢障碍和膜结构功能有密切关系。前文曾报导胞二磷胆碱(简称CDP胆碱)能抑制贮存红细胞的溶血,而且其作用主要不是通过能量代谢。本文试从CDP胆碱与红细胞膜磷脂的关系等方面作一些探索。将去除血浆的红细胞贮存于代血浆中,看到CDP胆碱仍能抑制溶血。经测定血浆磷脂的含量,也没有发现CDP胆碱血和对照之间有明显差别。此结果提示CDP胆碱抑制红细胞溶血的作用与血浆并无联系,排除了通过与血浆进行磷脂交换来更新红细胞磷脂,从而抑制溶血的可能性。此外,还测定了CDP胆碱血和对照的红细胞膜磷脂含量,并用薄板层析法测定了卵磷脂和神经鞘磷脂的百分含量。结果显示,实验组并不比对照有所增高,这也表明CDP胆碱对红细胞膜磷脂含量没有显著增进作用。除CDP胆碱以外,CMP也具有抑制贮存红细胞溶血的作用,其意义值得作进一步研究。     

Folding patterns of chromatin in chicken erythrocyte studied by using AFM and TEM

Four folding patterns (～15, ～30-40, ～60 and ～90-110 nm) of the higher_order structure of chromatin in chicken erythrocytes have been observed with an atomic force microscope (AFM) under near natural conditions. Results showed that the chromatin in the interphase nucleus of chicken erythrocyte was not in a disorganized form but represented a higher_order conformation. The thin sections of the chromatin in the nucleus of chicken erythrocyte were examined with a transmission electron microscope (TEM). Compared with TEM, AFM had the advantages that the samples did not need the preparation procedures such as fixing, sectioning and staining, and thus were closer to their native condition.     

Sodium-dependent uptake of inorganic phosphate by the intracellular malaria parasite

As the malaria parasite, Plasmodium falciparum, grows within its host erythrocyte it induces an increase in the permeability of the erythrocyte membrane to a range of low-molecular-mass solutes, including Na+ and K+ (ref. 1). This results in a progressive increase in the concentration of Na+ in the erythrocyte cytosol. The parasite cytosol has a relatively low Na+ concentration and there is therefore a large inward Na+ gradient across the parasite plasma membrane. Here we show that the parasite exploits the Na+ electrochemical gradient to energize the uptake of inorganic phosphate (P(i)), an essential nutrient. P(i) was taken up into the intracellular parasite by a Na+-dependent transporter, with a stoichiometry of 2Na+:1P(i) and with an apparent preference for the monovalent over the divalent form of P(i). A P(i) transporter (PfPiT) belonging to the PiT family was cloned from the parasite and localized to the parasite surface. Expression of PfPiT in Xenopus oocytes resulted in Na+-dependent P(i) uptake with characteristics similar to those observed for P(i) uptake in the parasite. This study provides new insight into the significance of the malaria-parasite-induced alteration of the ionic composition of its host cell.     

高压互感器在线故障监测透气装置的实验研究

实现了适用于１１０ｋＶ高压互感器的油气分离装置。用高分子膜进行了高压互感器油中溶解气体透气特性的大量实验研究。结果表明，高分子膜能将油中溶解气体分离。由于互感器结构的限制，安装部位是油不流动的区域，因此安装在互感器的气敏传感器要比安装在变压器的传感器灵敏度提高１０倍。监测装置的成本将相应提高。