红细胞膜弹性特性的AFM力曲线测量

人血红细胞生存环境的微小变化都会直接影响到膜骨架蛋白网络形变和细胞膜弹性力学性能的改变。利用原子力显微镜力曲线测量红细胞膜表面弹性,根据相关力。距离曲线图谱信息,分析活性红细胞在不同生理溶液环境中细胞膜弹性、黏附性与渗透脆性等变化特征。实验结果表明,经不同生理溶液制备的红细胞在维持其渗透压平衡条件下,细胞形态特征基本正常,直径约7-8μm。当力曲线图谱表征的膜弹性等力学特性,波动较大且差异明显时,形变恢复作用力变化范围为30-150pN,黏附力变化范围为3.9-35nN。尤其是在探针接近和撤离膜表面过程中,不同程度的多峰锯齿波、压痕深度、黏滞拖拽线程和链状伸展形式的相关力曲线图谱,表征细胞膜表面存在着复杂作用力。在无Ca^2＋／Mg^2＋的D-PBS缓冲液中,通过调节Na^＋,K^＋浓度,改变生理溶液渗透压,红细胞形变显著,出现棘轮状、球形和血影空囊等形态,对应的细胞膜弹性等力曲线图谱信息也相对更为复杂。研究表明红细胞膜弹性特性和膜骨架形变都与生理环境中的溶质（糖类与晶体电解质等）、浓度和渗透压等因素有直接关系。这为深入了解生物膜的结构和动力学提供了证据,也为原子力显微镜应用于红细胞膜病的临床诊断提供了依据。     

高HCY培养内皮细胞表面形态的AFM观察

目的：探讨一种新的损伤小、分辨率高的观察内皮细胞表面形态的方法。方法：取新生胎儿脐带静脉血管内皮细胞,培养２４～４８小时后,分为高ＨＣＹ实验组和正常对照组,采用ＡＦＭ对用戊二醛固定与未固定的内皮细胞进行观察,并与倒置显微镜、电子显微镜进行比较。结果：（１）固定后的内皮细胞ＡＦＭ扫描图象见细胞表面颗粒界限分明,排列规则,平均尺度为３００～４００ｎｍ,平整度略好于培养条件下观察的内皮细胞图象;（２）固定的高ＨＣＹ实验组内皮细胞的ＡＦＭ扫描图象见细胞表面有非常多的突起,颗粒规则性排列的特征消失,颗粒间界限模糊,表面粗糙,呈虫蚀状变化。（３）倒置显微镜观察未见两组间差异,电子显微镜观察也无法分辨差异。结论：ＡＦＭ是一种观察内皮细胞表面形态的较好方法,损伤小、分辨率高     

AFM对小鼠胚胎干细胞的形态学研究

目的：对已建系BALB/c小鼠胚胎干细胞膜表面进行纳米级超微结构的初步形态学研究,从而为从分子水平研究胚胎干细胞的增殖与分化调控机理奠定基础。方法：利用原子力显微镜（AFM）,在空气中对小鼠胚胎干细胞膜表面扫描成像。结果：AFM图像表现BALB/c小鼠胚胎干细胞呈圆盘状,直径约10-15μm,高约2-4μm,胚胎干细胞膜表面比较复杂,随着扫描范围的减小,切向分辨率逐渐增大,可达到纳米分辨,细胞表面有许多紧密堆积的椭球状颗粒。颗粒尺寸（x-y方向）为40-80nm。结果：利用AFM可以得到胚胎干细胞表面高分辨的,可重复的图像。     

神经节苷脂对细胞膜损伤的保护作用

观察了神经节苷脂对去垢剂Ｔｒｉｔｏｎ—Ｘ—１００损伤大鼠红细胞膜的影响。适量的红细胞与１６μｇ／ｍｌ至５×１０４μｇ／ｍｌ范围内不同浓度的神经节苷脂于３７℃孵育１小时后,加入ＴｒｉｔｏｎＸ－１００在室温下作用３分钟。离心,取上清液在５４０ｎｍ测溶液透光率,取沉淀部分的红细胞在扫描电镜下观察红细胞及其膜的形态变化。实验结果显示,在神经节苷脂浓度为１０４μ／ｍｌ时,对红细胞膜的保护作用最好,上清液透光率为正常值的８５．４％;细胞形态虽有改变,但仍的部分接近正常的细胞存在。而损伤对照组上清透光度仅为正常值的１３．４％,红细胞胞体缩小,成为棘形红细胞。结果表明,神经节苷脂对红细胞膜化学损伤有良好的保护作用。     

激光作用质粒DNA和小牛胸腺DNA的AFM研究

激光作用质粒ＤＮＡ和小牛胸腺ＤＮＡ产生损伤效应,导致ＤＮＡ结构变化,利用一种改进的试样制备过程和纳米显微镜--原子力显微镜（ＡＦＭ）能够获得可重现的激光作用质粒ＤＮＡ和小牛胸腺ＤＮＡ的ＡＦＭ图象,显示它们的特殊的表面结构。     

红芪、黄芪及配伍当归对环磷酰胺所致血虚模型小鼠的干预作用

目的：观察红芪、黄芪及配伍当归对环磷酰胺（CTX）所致血虚模型小鼠的干预作用。方法：昆明种小鼠随机分为7组,每组10只。采用CTX复制小鼠血虚模型,正常组与模型组小鼠用生理盐水灌胃,阳性对照组小鼠给予驴胶补血颗粒,四个给药组分别灌胃红芪、黄芪、红芪-当归组（5：1）及黄芪-当归组（5：1）,连续灌胃7 d后,采用血细胞分析仪测定各组小鼠外周红细胞（RBC）、淋巴细胞（LYM）、红细胞压积（HCT）、白细胞（WBC）、血小板（PLT）,取脾脏、胸腺、股骨,计算脾脏指数（SI）、胸腺指数（TI）的变化,进行网织红细胞计数（RC）、骨髓有核细胞计数,并对股骨进行病理切片观察。结果：与正常组比较,模型组小鼠RBC、WBC、HCT、PLT、LYM等含量均降低（P<0.05）,与模型组比较,红芪及不同配伍各组小鼠中SI、TI均显著升高（P<0.05）,红芪-当归组及黄芪-当归组可显著提高外周RBC、HCT、WBC、PLT、LYM的含量（P<0.05）,可升高RC及骨髓有核细胞数量（P<0.05）,股骨病理切片显示有所改善。结论：红芪-当归（5：1）与黄芪-当归（5：1）配伍对血虚模型小鼠的改善作用及补血作用优于红芪、黄芪的单独作用,黄芪-当归（5：1）效果更佳。     

型糖尿病患者红细胞膜中性糖、氨基糖及唾液酸含量的改变

 为阐明Ⅱ型糖尿病（ Ｎ Ｉ Ｄ Ｄ Ｍ ）患者红细胞膜的化学组成在非酶糖基化（ Ｎ Ｅ Ｇ）影响下发生的变化,采用毛细管气相色谱法和分光光度法测定了 ２０ 名正常人和 １９ 例Ⅱ型糖尿病患者红细胞膜的 ４ 种结合中性糖与 ２ 种氨基糖和唾液酸含量以及 ４ 种寡糖链上的末端中性糖和唾液酸含量．结果表明, Ｎ Ｉ Ｄ Ｄ Ｍ 患者红细胞膜几种结合单糖（ Ｇｌｃ, Ｆｕｃ, Ｇｌｃ Ａ, Ｇａｌ Ａ）与游离单糖（ Ｇｌｃ, Ｇａｌ, Ｍａｎ, Ｆｕｃ）含量以及唾液酸含量均较正常对照组明显降低（ Ｐ＜ ００１ 或 ００５）．据此推测,由于患者红细胞膜蛋白的重度糖基化导致某些膜结构蛋白的氧化损伤,细胞膜糖类含量的减少,可能是重度糖基化的继发后果．     

硫酸锌对肠系膜上动脉闭塞性休克后肺损伤的治疗作用

目的：探讨硫酸锌对肠系膜上动脉闭塞性（SMAO）休克后肺损伤的治疗作用及其机制。方法：行腹部手术分离出肠系膜上动脉并夹闭,2h后松夹,复制家兔SMAO休克模型。检测给与硫酸锌后SMAO休克家兔血浆及红细胞膜丙二醛（MDA）、红细胞超氧化物歧化酶（SOD）、血浆黄嘌呤氧化酶（XOD）含量;同时检测肺组织MDA、SOD和肺表面活性物质（PS）的含量,并观察小肠和肺的病理改变。结果：硫酸锌可使MDA及XOD含量降低,且可防止SOD和PS减少,避免了SMAO休克后肺损伤的进行性加剧。结论：SMAO休克并发急性肺损伤与组织氧化过程代谢紊乱有关,硫酸锌通过其稳膜作用能减轻或在一定程度上防止休克后肺损伤的发生和发展。     

超氧歧化酶(SOD)对人红细胞膜的保护作用

前文曾报道,红细胞膜经外源O₂^-_·自由基(碱性条件下,邻苯三酚自氧化产生)处理后有明显损伤作用。在此基础上,本文研究猪红细胞SOD对人红细胞膜氧化损伤的保护作用。结果表明,外源SOD对人红细胞膜有明显保护作用,表现为膜流动性和膜重封闭能力保持正常水平,膜蛋白交联作用有显著减小。     

超抗原刺激Jurkat T细胞的原子力显微镜研究

利用原子力显微镜分别对未加药人急性T淋巴细胞性白血病细胞（Jurkat细胞）和经超抗原刺激不同时间（6,12,48,72 h）后的Jurkat 细胞进行了细胞全貌和细胞膜表面纳米结构成像和探测研究,并通过比较不同状态下细胞表面的粘附力变化,探讨Jurkat细胞形态变化与粘附行为之间的关系,用CCK-8检测细胞的增殖,以期对Jurkat 细胞形态结构和细胞功能之间的关系有进一步的了解。结果发现：与未加药的Jurkat 细胞相比,随着超抗原刺激时间的延长,Jurkat 细胞的体积、高度、半宽度、粗糙度等参数发生明显的变化;活化48、72 h时,细胞与针尖间的相互作用力大约是活化6 h时的5倍,活化过程中细胞膜表面纳米结构的改变,引起其机械性能的变化。Jurkat 细胞的表面超微结构、细胞膜结构的改变和分化对于更深入地了解T细胞活化与免疫信号的传递,阐明其免疫过程的作用机制具有重要的意义。     

Tuning of electronic properties of one-dimensional cyano-bridged Cu-Ni, Cu-Pd, and Cu-Pt bimetallic assemblies by stereochemistry of ligands

Preparations, XPS and electronic spectroscopy, and magnetism of seven new one-dimensional cyano-bridged coordination polymers, chiral [Cu(RR-chxn)₂][Pd(CN)₄] · 2H₂O (1), [Cu(trans-chxn)₂][M(CN)₄] · 2H₂O (2, 4, and 6 for M = Pd, Ni, and Pt), and [Cu(cis-chxn)₂][M(CN)₄] · 2H₂O (3, 5, and 7 for M = Pd, Ni, and Pt) (RR-chxn = cyclohexane-(1R,2R)-diamine, trans-chxn = racemic trans-cyclohexane-(1,2)-diamine, and cis-chxn = racemic cis-cyclohexane-(1,2)-diamine) have been reported in view of tuning of their electronic properties by stereochemistry of chxn ligands and metal-substitution. Comparison of Cu 2p_1/2 and 2p_3/2 peaks of XPS and broad d-d bands around 18 000 cm⁻¹ of electronic spectra are described systematically for 1-7. Variable-temperature magnetic measurement shows that complexes 1-7 indicate weak antiferromagnetic interactions via cyano-bridges. Because of semi-coordination coupled with pseudo Jahn-Teller elongation and electrostatic interaction for 1, the axial Cu-N coordination bond distances of 2.330(7) and 3.092(8) Å are considerably longer than those of equatorial ones in the range from 2.016(6) to 2.030(6) Å. The former bond distances of 1 are intermediate values among the related Ni (2.324(6) and 3.120(8) Å) and Pt (2.34(1) and 3.09(1) Å) complexes.     

Land,livestock, and livelihoods: Changing dynamics of gender,caste, and ethnicity in a Nepalese Village

Over the past 10 years, Ghusel VDC, Lalitpur District has moved from primarily subsistence agriculture into the wider cash economy aided by the Small Farmers' Development Program (SFDP), which provides credit to farmers mainly for the purchase of buffalo for milk production, and by the National Dairy Corporation, which supports local dairy cooperatives. Analysis reveals that buffalo-keeping and milk sales are increasing the well-being of many households, while at the same time creating new inequalities in gender roles and responsibilities, greater inequities between Brahmin and Tamang residents in Ghusel, and placing pressures on the ecosystem for increased supplies of fodder and fuelwood. Evidence suggests that there is critical, need for attention to the social, and particularly gender-based, implications of maintaining livestock for milk sales and to the ecological underpinnings of this livelihood system.     

Structure and function of S-adenosylhomocysteine hydrolase

In mammals, S-adenosylhomocysteine hydrolase (AdoHcyase) is the only known enzyme to catalyze the breakdown of S-adenosylhomocysteine (AdoHcy) to homocysteine and adenosine. AdoHcy is the product of all adenosylmethionine (AdoMet)-dependent biological transmethylations. These reactions have a wide range of products, and are common in all facets of biometabolism. As a product inhibitor, elevated levels of AdoHcy suppress AdoMet-dependent transmethylations. Thus, AdoHcyase is a regulator of biological transmethylation in general. The three-dimensional structure of AdoHcyase complexed with reduced nicotinamide adenine dinucleotide phosphate (NADH) and the inhibitor (1′R, 2′S, 3′R)-9-(2′,3′-dihyroxycyclopenten-1-yl)adenine (DHCeA) was solved by a combination of the crystallographic direct methods program, SnB, to determine the selenium atom substructure and by treating the multiwavelength anomalous diffraction data as a special case of multiple isomorphous replacement. The enzyme architecture resembles that observed for NAD-dependent dehydrogenases, with the catalytic domain and the cofactor binding domain each containing a modified Rossmann fold. The two domains form a deep active site cleft containing the cofactor and bound inhibitor molecule. A comparison of the inhibitor complex of the human enzyme and the structure of the rat enzyme, solved without inhibitor, suggests that a 17° rigid body movement of the catalytic domain occurs upon inhibitor/substrate binding.     

Chiral Separation of G‐type Chemical Warfare Nerve Agents via Analytical Supercritical Fluid Chromatography

Chemical warfare nerve agents (CWNAs) are extremely toxic organophosphorus compounds that contain a chiral phosphorus center. Undirected synthesis of G‐type CWNAs produces stereoisomers of tabun, sarin, soman, and cyclosarin (GA, GB, GD, and GF, respectively). Analytical‐scale methods were developed using a supercritical fluid chromatography (SFC) system in tandem with a mass spectrometer for the separation, quantitation, and isolation of individual stereoisomers of GA, GB, GD, and GF. Screening various chiral stationary phases (CSPs) for the capacity to provide full baseline separation of the CWNAs revealed that a Regis WhelkO1 (SS) column was capable of separating the enantiomers of GA, GB, and GF, with elution of the P(+) enantiomer preceding elution of the corresponding P(–) enantiomer; two WhelkO1 (SS) columns had to be connected in series to achieve complete baseline resolution. The four diastereomers of GD were also resolved using two tandem WhelkO1 (SS) columns, with complete baseline separation of the two P(+) epimers. A single WhelkO1 (RR) column with inverse stereochemistry resulted in baseline separation of the GD P(–) epimers. The analytical methods described can be scaled to allow isolation of individual stereoisomers to assist in screening and development of countermeasures to organophosphorus nerve agents. Chirality 26:817–824, 2014. © 2014 The Authors. Chirality published by John Wiley Periodicals, Inc.     

Influence of Fluoride on Rat Kidney Antioxidant System: Effects of Methionine and Vitamin E

The aim of the study has been to determine and compare the influence upon the kidney antioxidative system, exercised by administration of vitamin E, and vitamin E in combination with methionine, under conditions of oxidative stress induced by sodium fluoride. The experiment was carried out on Wistar FL rats (adult males) that, for 35 days, were administered water, NaF, NaF with vitamin E, or vitamin E with methionine (doses: 10 mg NaF/kg of body mass/24 h, 3 mg vitamin E per 10 μl per rat for 24 h, 2 mg methionine per rat for 24 h). The influence of administered sodium fluoride and antioxidants upon the antioxidative system in kidney was examined by analyzing the concentration of malondialdehyde (MDA) and the activity of the most important antioxidative enzymes (SOD, total and both its isoenzymes, GPX, GST, GR, and CAT). The studies carried out confirmed the disadvantageous effect of the administered dose of NaF upon the antixodiative system in rats (increase in the concentration MDA, decrease activity of all antioxidative enzymes). The administration of vitamin E increased the activity of studied enzymes with the exception of glutathione reductase GR; it also reduced the procesess of lipid peroxidation. It has been found that combined doses of vitamin E and methionine were most effective in inhibiting lipid peroxidation processes. The results confirmed the antioxidative properties of methionine.     

Reorientation of Microfibrils and Microtubules at the Outer Epidermal Wall of Maize Coleoptiles During Auxin-Mediated Growth

Auxin-mediated elongation growth of isolated subapical coleoptile segments of maize (Zea mays L.) is controlled by the extensibility of the outer cell wall of the outer epidermis (Kutschera et al., 1987). Here we investigate the hypothesis that auxin controls the extensibility of this wall by changing the orientation of newly deposited microfibrils through a corresponding change in the orientation of cortical microtubules. On the basis of electron micrographs it is shown that cessation of growth after removal of the endogenous source of auxin is correlated with a relative increase of longitudinally orientated microfibrils and microtubules at the inner wall surface. Conversely, reinduction of growth by exogenous auxin is correlated with a relative increase of transversely orientated microfibrils and microtubules at the inner wall surface. These changes can be detected 30–60 min after the removal and addition of auxin, respectively. The functional significance of directional changes of newly desposited wall microfibrils for the control of elongation growth is discussed.     

Gamma-glutamylcysteine protects ergothioneine-deficient Mycobacterium tuberculosis mutants against oxidative and nitrosative stress

Mycobacterium tuberculosis (M.tb.), the causative agent of tuberculosis (TB), cannot synthesize GSH, but synthesizes two major low molecular weight thiols namely mycothiol (MSH) and ergothioneine (ERG). Gamma-glutamylcysteine (GGC), an intermediate in GSH synthesis, has been implicated in the protection of lactic acid bacteria from oxidative stress in the absence of GSH. In mycobacteria, GGC is an intermediate in ERG biosynthesis, and its formation is catalysed by EgtA (GshA). GGC is subsequently used by EgtB in the formation of hercynine-sulphoxide-GGC. In this study, M.tb. mutants harbouring unmarked, in-frame deletions in each of the fives genes involved in ERG biosynthesis (egtA, egtB, egtC, egtD and egtE) or a marked deletion of the mshA gene (required for MSH biosynthesis) were generated. Liquid chromatography tandem mass spectrometry analyses (LC-MS) revealed that the production of GGC was elevated in the MSH-deficient and the ERG-deficient mutants. The ERG-deficient ΔegtB mutant which accumulated GGC was more resistant to oxidative and nitrosative stress than the ERG-deficient, GGC-deficient ΔegtA mutant. This implicates GGC in the detoxification of reactive oxygen and nitrogen species in M.tb.     

In vitro studies of the rabbit immune system. IV. Differential mitogen responses of isolated T and B cells.

The mitogenic responses of separated rabbit lymphocyte populations functionally analogous to mouse T and B cells have been tested in vitro. Purified T cells were prepared by passage over nylon wool (NW) and purified B cells prepared by treatment with antithymocyte serum and complement (ATS + C). ATS + C kills 70% of peripheral blood lymphocytes (PBL's) and 50% of the spleen cells while passage over NW yields 40% of the applied PBL's and 5–23% of the applied spleen cells. NW-purified T cells from the spleen or PBL's respond fully to concanavalin A (Con A) but have a reduced response to phytohemaglutinin (PHA) and little or no response to goat anti-rabbit immunoglobulin (anti-Ig). PBL's that survive ATS + C (B cells) are stimulated by anti-Ig but not by Con A or PHA. B cells purified from spleen do not respond to Con A or PHA but will respond to anti-Ig under appropriate conditions. A full spleen B-cell response to anti-Ig required removal of Ig produced by the cultures that blocked anti-Ig stimulation. It is concluded that, for rabbit lymphocytes, Con A and PHA are primarily T-cell mitogens and that anti-Ig is primarily a B-cell mitogen. However, the mitogen response of unfractionated PBL or spleen cell populations indicates an overlap in reactivity. This could be due to cells sharing T and B properties, alteration of cell populations by the fractionation procedures used, or recruitment of one population in the presence of a mitogenic response of the other population.     

八种脑-肠肽侧脑室内注射对大鼠基础胃酸分泌的影响

用乌拉坦麻醉大鼠作急性实验,采用连续灌流胃并收集流出液的方法,观察向侧脑室内注射微量脑-肠肽对大鼠基础胃酸分泌的影响。实验结果如下:(1)雨蛙肽、八肽胆囊收缩素、促甲状腺素释放激素及四肽胃泌素均使总酸排出量增加;(2)生长抑素、胰多肽、P 物质、胰高血糖素则使总酸排出量减少;(3)上述肽类用侧脑室注射的剂量作肌肉注射,除四肽胃泌素也产生明显的刺激胃酸分泌作用外,对胃酸分泌均无明显影响。以上结果提示,脑内的一些肽类可能以神经递质或调制物的方式,参与中枢对胃酸分泌的调节。     

A chromium-tolerant plant growing in Cr-contaminated land

In this paper, a kind of Typhaceae plant species, Typha angustifolia L, with a high tolerance to Cr is described. Experiments were carried out to examine its ability to tolerant Cr and its physiological response. The results showed that there was no difference in growth, plant height, and biomass response to external Cr (VI) between the plants exposed to 100 microM Cr (VI) and control (0 microM), while increasing Cr levels to 200-800 microM induced a significant decrease in plant height and biomass, but no significant injury was detected, even for the plants exposed to 800 microM Cr. Chromium induced significant increases in superoxide dismutase (SOD) and peroxidase (POD) activities. Meanwhile, a significantly positive correlation was found between Cr and Mn or Cu in leaves and roots, respectively. The Cr tolerance of the plant appeared to be associated with the enhancement of SOD and POD activities and the improvement in uptake and translocation of the essential microelements.