羧甲基壳聚糖膜对皮肤成纤维细胞相容性研究

通过玻璃流延法制备不同分子质量的壳聚糖及羧甲基壳聚糖膜,以体外培养的人皮肤成纤维细胞作为对象,利用膜的浸渍液培养及膜表面直接培养法研究比较了两种多糖膜的细胞相容性.实验发现两种多糖膜的浸渍液对细胞均无毒性效应,生物安全性是小分子质量膜好于大分子质量膜,羧甲基壳聚糖膜好于壳聚糖膜.皮肤成纤维细胞在壳聚糖膜上的生长受到抑制,生长一段时间后细胞有聚集和脱落现象,而羧甲基壳聚糖膜上细胞能很好地贴附、生长,没有聚集和脱落现象.结果表明羧甲基壳聚糖膜具有比壳聚糖膜更优越的细胞相容性.     

壳聚糖膜在治疗宫颈疾病中的降解时间与影响因素的研究

目的:研究不同壳聚糖膜在体外及体内的生物降解时间。方法:用不同分子量壳聚糖为制膜材料,将4组不同组成成分的壳聚糖溶于1.5%乙酸中,配成1%溶液,加入辅料烘干成膜称其重量,剪成大小相等的小块,精称后放置于弱酸介质和雌鼠阴道中。定时取样,水洗,烘干后称重,按W-W'W×100%算得降解百分率。结果:1号较2号降解时间快,而加入PVA的3号与4号又较1号与2号更快。结论:壳聚糖是一种生物可降解材料,壳聚糖膜的降解时间与其分子量、PVA比例、介质酸性强度等因素相关。     

壳聚糖在给药系统中的应用

壳聚糖来源丰富,具有良好的生物相容性、生物可降解性、无毒性、成膜性和极强的可塑性,已经作为高分子材料,广泛用于给药系统中.将壳聚糖应用于给药系统中可以提高药物安全性、有效性及可靠性,可以调整药物释放速率,减少给药次数.此外,壳聚糖可塑性强,可制成膜、压成片、制成颗粒、微球或增粘剂等多种剂型.该文就壳聚糖的特性及其在给药系统中的应用予以综述.     

壳聚糖/ 魔芋葡甘露聚糖复合膜体内外促创面愈合研究

目的:制备壳聚糖/魔芋葡甘露聚糖复合膜,研究其促创面愈合作用。方法:壳聚糖溶液和魔芋葡甘露聚糖溶液混合后冷冻干燥制成复合膜。扫描电镜观察膜的形态和孔径,并研究比较膜的吸水率,水蒸气透过率,拉伸强度,断裂伸长率和体外降解率。建立大鼠皮肤损伤模型,敷以复合膜治疗,比较创面愈合率,观察创面组织染色结果,评价复合膜的促创面愈合作用。结果:壳聚糖/魔芋葡甘露聚糖复合膜具有三维网状结构,壳聚糖复合魔芋葡甘露聚糖后,膜的吸水率、拉伸强度和断裂伸长率提高,体外降解加速,水蒸气透过率改善。愈合实验表明壳聚糖/魔芋葡甘露聚糖膜具有促进创面愈合作用。结论:壳聚糖/魔芋葡甘露聚糖复合膜制备工艺简单,能有效促进创面愈合,具有成为创伤敷料的潜力。     

壳聚糖基角膜细胞载体的制备及其细胞相容性

为探讨羟丙基壳聚糖基共混膜作为组织工程技术中角膜细胞培养载体的可行性, 分别制备了羟丙基壳聚糖/硫酸软骨素、羟丙基壳聚糖/明胶/硫酸软骨素以及羟丙基壳聚糖/氧化透明质酸/硫酸软骨素三种共混膜。测定其透光率、含水量和蛋白吸附性能; 在共混膜上培养兔角膜上皮细胞, 通过观察角膜上皮细胞在不同载体膜上的生长状态、贴附情况, 测定细胞活性以及上清液中乳酸脱氢酶的活性, 研究三种壳聚糖基载体膜片与角膜上皮细胞的相容性。膜片理化性质测定结果表明三种共混膜片具有良好的透明度, 适宜的含水量和较强的蛋白吸附性能; 细胞相容性实验结果表明羟丙基壳聚糖/明胶/硫酸软骨素共混膜对细胞的损伤最小, 有利于细胞在膜上的贴附和生长, 表现出良好的细胞相容性, 有望作为角膜细胞载体体外构建组织工程化角膜。     

丙烯酰胺均相改性甲壳素及共混膜研究

低温下将甲壳素直接溶于8%氢氧化钠/4%尿素水溶液,以丙烯酰胺为醚化剂,均相合成丙烯酰胺改性甲壳素(AMC)。产物用~~1H NMR和FT-IR进行了表征,并初步研究了其稀溶液的粘度行为。然后采用相转化法制备了AMC/壳聚糖共混膜,研究了膜的力学性能、吸水率、透光率及对大肠杆菌和金黄色葡萄球菌的抑菌性能。结果表明:Na OH/尿素水体系下成功均相制备了AMC,其取代度为0.49,水溶性好,AMC稀溶液表现出典型的聚电解质行为;AMC和壳聚糖两种高分子具有较好的相容性,AMC的加入明显改善了壳聚糖膜的拉伸强度和抗水性,且所有的共混膜均有显著的抑菌效果。     

壳聚糖絮凝纯化香菇多糖的研究

研究了壳聚糖对香菇多糖提取液的沉降作用,考察了壳聚糖用量、沉降时间、溶液酸度对吸附色素、沉降蛋白质和多糖损失的影响,同时考察了壳聚糖对超滤膜分离初始膜通量的影响。实验结果表明壳聚糖对香菇多糖提取液中色素和蛋白质具有较好的絮凝沉降效果,当壳聚糖的用量为5.0mg/mL,絮凝沉降时间为40min,溶液pH为5.0时,60r/min的转速搅拌条件下,色素含量降低了约28%,蛋白质含量降低了42%左右,而多糖含量只损失8.9%左右,溶液粘度降低了11.8%,采用超滤分离时,经壳聚糖沉降的多糖溶液初始膜通量增加了14%左右。     

壳聚糖被膜对鲜切荸荠褐变的抑制

壳聚糖被膜抑制鲜切荸荠切片的苯丙氨酸解氨酶、多酚氧化酶、过氧化物酶的活性 ,延缓切片的褐变 ,保持荸荠的食用品质 ,减少腐烂。在 0 .5 %～ 2 %范围内 ,壳聚糖被膜对荸荠褐变的抑制效应随使用浓度的增加而增强。     

壳聚糖反渗透膜的制备初探

本文介绍了从虾、蟹甲壳制取壳聚糖用于制备反渗透膜的方法。运用正交试验设计分析制膜条件对膜性能的影响。     

通过研究改性壳聚糖与细胞的相互作用评价其生物相容性

利用细胞生物学的方法, 研究了四种不同的细胞在经过改性的壳聚糖（ＣＨＩＴＯＳＡＮ） 膜上的生长,测定了细胞相对黏附力、细胞初始黏附率, 并利用ＦＤＡ 实验测定了细胞活力,从而从多个方面评价了这几种不同材料的生物相容性。实验结果表明,与明胶交联的壳聚糖膜明显比其它两种膜有利于细胞的黏附和生长,为进一步对材料进行筛选奠定了基础。     

Probing membrane potential with nonlinear optics.

The nonlinear optical phenomenon of second harmonic generation is shown to have intrinsic sensitivity to the voltage across a biological membrane. Our results demonstrate that this second order nonlinear optical process can be used to monitor membrane voltage with excellent signal to noise and other crucial advantages. These advantages suggest extensive use of this novel approach as an important new tool in elucidating membrane potential changes in biological systems. For this first demonstration of the effect we use a chiral styryl dye which exhibits gigantic second harmonic signals. Possible mechanisms of the voltage dependence of the second harmonic signal are discussed.     

Studying the Nucleated Mammalian Cell Membrane by Single Molecule Approaches

The cell membrane plays a key role in compartmentalization, nutrient transportation and signal transduction, while the pattern of protein distribution at both cytoplasmic and ectoplasmic sides of the cell membrane remains elusive. Using a combination of single-molecule techniques, including atomic force microscopy (AFM), single molecule force spectroscopy (SMFS) and stochastic optical reconstruction microscopy (STORM), to study the structure of nucleated cell membranes, we found that (1) proteins at the ectoplasmic side of the cell membrane form a dense protein layer (4 nm) on top of a lipid bilayer; (2) proteins aggregate to form islands evenly dispersed at the cytoplasmic side of the cell membrane with a height of about 10–12 nm; (3) cholesterol-enriched domains exist within the cell membrane; (4) carbohydrates stay in microdomains at the ectoplasmic side; and (5) exposed amino groups are asymmetrically distributed on both sides. Based on these observations, we proposed a Protein Layer-Lipid-Protein Island (PLLPI) model, to provide a better understanding of cell membrane structure, membrane trafficking and viral fusion mechanisms.     

Vacuolar ethylene formation does not depend on membrane potential

Enzymatic synthesis of ethylene in the vacuole is assumed to require membrane integrity. The possibility that this reflects dependence on the vacuolar membrane potential was investigated. Vacuoles were released from protoplasts isolated from leaves of Vicia faba L. cv. Cyprus. The dependence of the ethylene-forming activity on tonoplast integrity was re-examined by immobilization of the vacuoles in a cross-linked polymeric matrix and subsequent permeabilization of the tonoplast with toluene, a pore-forming reagent. The relationship between the vacuolar ethylene formation and the membrane potential of free vacuoles was investigated by following the uptake of thiocyanate using permeabilized, depolarized and hyperpolarized vacuoles. Toluene and the proton conductor carbonyl cyanide m -chlorophenylhydrazone (CCCP) caused loss of ethylene-forming activity and depolarized the vacuolar membrane potential. However, depolarization of the membrane potential with choline chloride and hyperpolarization by ATP did not affect ethylene biosynthesis. These conflicting results lead to the conclusion that vacuolar ethylene biosynthesis is not dependent on the vacuolar membrane potential. The possibility that the inhibition of ethylene biosynthesis by toluene and CCCP may result from direct hydrophobic interactions between these compounds and hydrophobic components of the ethylene-forming enzyme is discussed.     

Calcium-binding protein of the chick chorioallantoic membrane. I. Immunohistochemical localization

The preparation of a specific antiserum (anti-CaBP) against the calcium-binding protein (CaBP) of the chorioallantoic membrane (CAM) is described. The anti-CaBP appeared to be specific for the CaBP by immunodiffusion and immunoelectrophoresis. Application of the anti-CaBP in immunofluorescence histochemistry revealed that the CaBP is present in the CAM only at developmental ages corresponding with the expression of the calcium transport function of the membrane. Furthermore, the CaBP is localized to the ectoderm of the CAM, appears to be exposed to the entire external surface of the ectoderm, and can be shown to be associated with cells enzymatically dissociated from the CAM. These results are consistent with a functional role of the CaBP in the CAM calcium transport process.     

KB-R7943, a plasma membrane Na/Ca exchanger inhibitor,blocks opening of the mitochondrial permeability transition pore

The isothiourea derivative, KB-R7943, inhibits the reverse-mode of the plasma membrane sodium/calcium exchanger and protects against ischemia/reperfusion injury. The mechanism through which KB-R7943 confers protection, however, remains controversial. Recently, KB-R7943 has been shown to inhibit mitochondrial calcium uptake and matrix overload, which may contribute to its protective effects. While using KB-R7943 for this purpose, we find here no evidence that KB-R7943 directly blocks mitochondrial calcium uptake. Rather, we find that KB-R7943 inhibits opening of the mitochondrial permeability transition pore in permeabilized cells and isolated liver mitochondria. Furthermore, we find that this observation correlates with protection against calcium ionophore-induced mitochondrial membrane potential depolarization and cell death, without detrimental effects to basal mitochondrial membrane potential or complex I-dependent mitochondrial respiration. Our data reveal another mechanism through which KB-R7943 may protect against calcium-induced injury, as well as a novel means to inhibit the mitochondrial permeability transition pore.     

The novel C-terminal KCNQ1 mutation M520R alters protein trafficking

The long QT-syndrome is characterized by a prolongation of the QT-interval and tachyarrhythmias causing syncopes and sudden death. We identified the missense mutation M520R in the calmodulin binding domain of the Kv7.1 channel from a German family with long QT-syndrome. Heterologous expression of the mutant did not reveal any whole-cell currents independent of the auxiliary subunit KCNE1. Co-expression of the wild-type Kv7.1 channels and the mutant showed that the mutant did not have a dominant negative effect. In immunocytochemical assays of transfected COS-1 cells wild-type Kv7.1 showed an immunopositive labeling of the plasma membrane. For M520R no plasma membrane staining was visible, instead a strong signal in the ER was observed. These results indicate that the LQT1 mutation M520R leads to ER-retention and dysfunctional trafficking of the mutant channel resulting in haploinsufficiency.     

A σW-dependent stress response in Bacillus subtilis that reduces membrane fluidity

Bacteria respond to physical and chemical stresses that affect the integrity of the cell wall and membrane by activating an intricate cell envelope stress response. The ability of cells to regulate the biophysical properties of the membrane by adjusting fatty acid composition is known as homeoviscous adaptation. Here, we identify a homeoviscous adaptation mechanism in Bacillus subtilis regulated by the extracytoplasmic function σ factor σ(W). Cell envelope active compounds, including detergents, activate a sense-oriented, σ(W)-dependent promoter within the first gene of the fabHa fabF operon. Activation leads to a decrease in the amount of FabHa coupled with an increase in FabF, the initiation and elongation condensing enzymes of fatty acid biosynthesis respectively. Downregulation of FabHa results in an increased reliance on the FabHb paralogue leading to a greater proportion of straight chain fatty acids in the membrane, and the upregulation of FabF increases the average fatty acid chain length. The net effect is to reduce membrane fluidity. The inactivation of the σ(W)-dependent promoter within fabHa increased sensitivity to detergents and to antimicrobial compounds produced by other Bacillus spp. Thus, the σ(W) stress response provides a mechanism to conditionally decrease membrane fluidity through the opposed regulation of FabHa and FabF.     

NaCl、Na2 SO4 和Na2GO3 胁迫对小麦叶片自由基含量及质膜透性的比较研究

 分别用浓度为25mmol／L、50mmol／L、100 mmol／L和200 MMOL／l的Nacl、Na2SO4和Na2CO3的营养液培养小麦4d,较之不含盐的营养液,其自由基含量上升,产生速率增加,叶片质膜透性增加。不同盐的影响也不同,在低浓度时,NaCl的影响大于Na2SO4,高浓度时,NaCl影响小于Na2CO3,Na2CO3的影响最为显著。实验结果也表现出小麦叶片自由基含量和质膜透性呈现较好的相关性。因此可认为,盐胁迫促使自由基含量增加,自由基通过过氧化作用影响质膜透性,从而影响植物的生长。     

Structural and immunochemical homogeneity of Aeromonas salmonicida lipopolysaccharide.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyze the lipopolysaccharides of typical and atypical strains of the fish pathogen Aeromonas salmonicida. 32P intrinsically radiolabeled lipopolysaccharide in sarcosinate-extracted outer membrane preparations, lipopolysaccharide stained by silver in proteinase K-digested outer membrane preparations and whole cell lysates, as well as purified lipopolysaccharide, displayed O-polysaccharide chains which were unusually homogeneous with respect to chain length. Chemical analysis further revealed that the sugar composition of the smooth lipopolysaccharide purified from three typical strains was very similar. Immunoblotting and immunofluorescent staining with both polyclonal and monoclonal antibody showed that the O-polysaccharide chains were strongly immunogenic and were antigenically cross-reactive on typical and atypical strains from diverse origins. Immunofluorescence analysis and phage binding studies demonstrated that a number of these O-polysaccharide chains traversed the surface protein array of virulent strains of A. salmonicida and were exposed on the cell surface.     

Effect of 3,5,3'-triiodo-L-thyronine on amino acid accumulation and membrane potential in Sertoli cells of the rat testis

The purpose of this study was to investigate the effect of T3 on amino acid accumulation and on the membrane potential of Sertoli cells of immature rat testes. Testes of pre-pubertal and pubertal rats were pre-incubated (30 min) in Krebs-Ringer bicarbonate buffer and incubated in the presence of [14C]methylaminoisobutyric acid with and without T3 for 15, 45 and 60 min. The hormone (10(-6) M and 10(-7) M) significantly stimulated amino acid accumulation in 6 and 13-day old rat testes but did not have any effect in neonatal and pubertal animals. T3 produced a dose-dependent hyperpolarizing effect at concentrations of 10(-6) M, 10(-5) M, 2 x 10(-5) M and 10(-4) M. We conclude that T3 induces a membrane hyperpolarization in Sertoli cells and stimulates amino acid accumulation in immature rat testes, demonstrating that the hormone has a rapid plasma membrane action.