壳聚糖锌离子固定化亲和层析填料的制备与性质

目的:制备了壳聚糖Zn2+固定化亲和层析填料,并对其性能进行了研究。方法:采用反相悬浮法制备了交联壳聚糖;再以环氧氯丙烷为活化剂,乙二胺为螯合配基,制备了固定化亲和层析填料;表征了其有效粒径以及均匀系数、含水量、失重率、氨基含量、骨架密度、堆积密度以及孔度值。从时间、加入ZnCl2的浓度、温度、pH方面对Zn2+固定化条件进行了优选,并确定了Zn2+的固定化量。含组氨酸标签的乙醛脱氢酶粗酶液,经硫酸铵盐析后,考察了壳聚糖Zn2+固定化亲和层析填料的亲和性能。结果:制备的填料有效粒径为105μm;均匀系数为1.46;含水量为58.03%;失重率为85.43%;氨基含量为9.20mmol/g;骨架密度为1.217 8g/ml;堆积密度为0.843 2g/ml;孔度值为36.40%。固定化Zn2+的最佳条件是:时间3 h、加入ZnCl2溶液浓度0.1mol/L、温度28℃、pH 5.5;且此条件下,亲和层析填料中Zn2+固定化量为3.35mmol/g。壳聚糖Zn2+固定化亲和层析填料对乙醛脱氢酶的亲和性能为4.14IU/g(干重)。结论:制备了壳聚糖Zn2+固定化亲和层析填料,可用于带有组氨酸标签重组蛋白的快速分离与纯化。     

DNA complexes with polycations useful for delivery of DNA into cells

Generation and physicochemical properties of complexes formed by high-molecular thymus DNA and plasmid DNA with synthetic polymers of (dimethyl amino)ethyl methacrylate, (diethyl amino)ethyl methacrylate, and poly(vinyl amine) were studied in solutions of different ionic strength using low-gradient viscometry, electrophoresis, circular dichroism, spectrophotometry, and dynamic light scattering. The complexes were tested for toxicity with T98G cell cultures. Condensation of DNA was shown to occur when the ratio of charged groups in the polycations and DNA exceeded unity. This condensation manifested itself as an increase in the optical density of DNA solutions. Condensation-associated changes in the dimensions of DNA molecules were determined, and phase diagrams of DNA-polycation systems were analyzed in the presence of NaCl. MTT analysis revealed no toxicity of these complexes.     

Conformational change from antiparallel beta-sheet to alpha-helix in a series of depsipeptide, -(Leu-Leu-Lac)(n)-: syntheses, spectroscopic studies, and crystal structures of Boc-Leu-Lac-OEt and Boc-(Leu-Leu-Lac)(n)-OEt (n = 1, 2)

The depsipeptides Boc-Leu-Lac-OEt (1) and Boc-(Leu-Leu-Lac)(n)-OEt (n = 1, 2) (2 and 3, respectively) (Boc = tert-butyloxycarbonyl, Lac = L-lactic acid residue) has been synthesized and studied by crystallographic, CD spectroscopic, and ESI-MS analyses. In the packing cells, those three compounds adopt beta-strand conformations. Each molecule is linked into a dimer (1) or an infinite assembly (2 and 3) by tight hydrogen bonds of the type NH...O==C. Interestingly, the hexamer, 3 shows the first example of antiparallel pleated beta-sheet crystal structure for a depsipeptide molecule. In the packing cells, especially for 3, the ester groups O--C==O are perpendicularly oriented to the amide groups NH--C==O and beta-sheet planes to avoid the interaction between --O--(ester) and O==C. Therefore, when the chain length become longer, the O...O==C repulsion interaction works as a beta-sheet breaker and hence promotes an alpha-helical structure as observed for Boc-(Leu-Leu-Lac)(3)-Leu-Leu-OEt (4) (Oku et al. Biopolymers 2004, 75, 242-254) and Boc-(Leu-Leu-Lac)(n)-OEt (n = 4-6) (5-7) (Katakai et al., Biopolymers 1996, 38, 285-290), in which the O...O==C repulsion does not cause significant structural changes in alpha-helical main chains. Therefore from the structural and spectroscopic analyses, we have found governing factors for the specificity in the beta-sheet and alpha-helix decision in this series of depsipeptides, -(Leu-Leu-Lac)(n)-.     

紫花苜蓿的花蜜量和访花蜜蜂数量对种子产量的影响

对10个紫花苜蓿品种的花萼直径、花蜜量、访花蜜蜂数量和种子产量进行了研究,结果表明,紫花苜蓿的单位面积花蜜量与访花蜜蜂数量呈极显著正相关(r=0.93**),访花蜜蜂数量与种子产量呈显著正相关(r=0.87*);紫花苜蓿的花蜜量与花萼直径呈极显著正相关(r=0.99**);紫花苜蓿品种赛特(Sitel)和德宝(Derby)在兰州地区能获得高产、优质的种子。     

光合性能选种法研究

生物良种的选择,历来是人类最关心的事,本文讨论的是植物的选种方法,我们在品质基本相同的几个品种中选择最优品种.这里最优品种指的就是产量最高者.植物的产量是由光合作用所产生的生物量,所以,光合性能强的物种常为最优品种.本文所介绍的选种方法,就是根据不同品种的光合性能的强弱来研究品种的优劣,而植物光合性能强弱可以通过植物的光合总量来确定.本文计算了三个猕猴桃品种的光合总量,其中光合总量最高者为最优品种（产量最高）,这与生产实践完全一致,所以光合性能选种法得到了生产实践的验证.     

NaCl胁迫下AM真菌对棉花生长和叶片保护酶系统的影响

利用盆栽实验研究了 Na Cl胁迫条件下 AM真菌对棉花生长和叶片保护酶系统的影响。结果表明 :在土壤中加入 0、0 .1%、0 .2 %、0 .3%浓度 Na Cl条件下 ,Na Cl胁迫对 AM真菌的接种效果有显著影响。接种 AM真菌提高了棉花根系菌根侵染率 ,增加了棉株的生物产量 ,以 0～ 0 .2 % Na Cl浓度时 AM真菌接种效果最好。 AM真菌对棉株生理参数和保护酶活性的影响因生育期和 Na Cl浓度不同而异 ,现蕾期和低盐浓度 (0～ 0 .1% )下叶片叶绿素含量明显增加 ;中高盐水平 (0 .2 %～ 0 .3% )和生育后期叶片可溶性蛋白质含量和 SOD、POD、CAT等保护酶活性显著提高 ,MDA含量明显降低 ;棉株 K、Ca、Mg含量因植株部位和盐浓度不同而变化。 AM真菌增强宿主植物的耐盐性可能源于促进宿主根系对土壤矿质元素吸收的直接作用和改善植物体内离子平衡和生理代谢活动、提高保护酶活性的间接作用     

The Food Amount Rating Scale: Development,Reliability, and Validity

Objective: Implied in measures of binge eating is the assumption that individuals agree on what comprises a large amount of food. However, whether individuals estimate food amounts similarly or whether estimation of food amounts varies as a function of personal characteristics is unknown. The Food Amount Rating Scale (FARS) is a standardized set of stimuli for assessing individuals’ judgment of food amounts. Research Methods and Procedures: Two versions of the FARS were developed, and their psychometric properties were assessed. These versions are the same in all respects except that the rater is instructed to rate various food amounts for the average woman on Form W and for the average man on Form M. Results: Content validity was confirmed by 14 researchers and research assistants in the field of eating disorders. The FARS is a 24-item inventory with adequate test–retest reliability (Form W = 0.85; Form M = 0.87) and split–half reliability (Form W = 0.90; Form M = 0.89). Convergent validity is suggested by the finding that ratings for the average woman (Form W) were significantly higher than ratings for the average man (Form M). Discussion: The FARS is a psychometrically sound tool for use in basic research focused on identifying whether the subjective judgment of food amounts varies as a function of personal characteristics and in clinical research where it may be important to know how individuals judge food amounts.     

四个与小鼠摄食、摄水量及蛋白质代谢相关基因的功能研究

目的　通过对小鼠摄食、摄水量及蛋白质代谢相关基因的功能研究 ,为今后的营养学和营养疾病学的分子生物学水平研究提供一定的依据。方法　利用反义核酸技术与动物行为学实验方法相结合 ,从动物整体水平上研究与动物膳食和营养代谢相关的基因功能。从课题组以往研究得到的摄食量与对照组有显著差异的基因中选出 4个 ,用BALB c小鼠进行实验。结果　实验结果表明 :D1、Pe和Tr实验组小鼠的摄食量、摄水量与对照组有明显的差异 ,D1和Tr实验组的蛋白质代谢与对照组有明显的差异。D1实验组小鼠的摄食量和摄水量明显高于对照组并呈下降趋势 ,但其表观消化率小于对照组 ,说明其蛋白质代谢功能有所下降 ;Tr实验组小鼠的摄食量和摄水量也明显高于对照组但呈上升趋势 ,其表观消化率大于对照组 ,说明其蛋白质代谢功能有所上升。结论　预测D1基因可能与促进营养代谢功能有关 ,Tr基因可能与抑制营养代谢功能有关。D1、Pe和Tr与移动、痛觉、记忆等其他行为学有相关性 ,P3与其他行为学无相关性。     

Direct anabolic metabolism of three-carbon propionate to a six-carbon metabolite occurs in vivo across tissues and species

Anabolic metabolism of carbon in mammals is mediated via the one- and two-carbon carriers S-adenosyl methionine and acetyl-coenzyme A. In contrast, anabolic metabolism of three-carbon units via propionate has not been shown to extensively occur. Mammals are primarily thought to oxidize the three-carbon short chain fatty acid propionate by shunting propionyl-CoA to succinyl-CoA for entry into the TCA cycle. Here, we found that this may not be absolute as, in mammals, one nonoxidative fate of propionyl-CoA is to condense to two three-carbon units into a six-carbon trans-2-methyl-2-pentenoyl-CoA (2M2PE-CoA). We confirmed this reaction pathway using purified protein extracts provided limited substrates and verified the product via LC-MS using a synthetic standard. In whole-body in vivo stable isotope tracing following infusion of ¹³C-labeled valine at steady state, 2M2PE-CoA was found to form via propionyl-CoA in multiple murine tissues, including heart, kidney, and to a lesser degree, in brown adipose tissue, liver, and tibialis anterior muscle. Using ex vivo isotope tracing, we found that 2M2PE-CoA also formed in human myocardial tissue incubated with propionate to a limited extent. While the complete enzymology of this pathway remains to be elucidated, these results confirm the in vivo existence of at least one anabolic three- to six-carbon reaction conserved in humans and mice that utilizes propionate.     

Field Hypothesis on the Self-regulation of Gene Expression

The mechanism of the self-regulation of gene expression in living cells is generally explained by considering complicated networks of key-lock relationships, and in fact there is a large body of evidence on a hugenumber of key-lock relationships. However, in the present article we stress that with the network hypothesis alone it is impossible to fully explain the mechanism of self-regulation in life. Recently, it has been established that individual giant DNA molecules, larger than several tens of kilo base pairs, undergo a large discrete transition in their higher-order structure. It has become clear that nonspecific weak interactions with various chemicals, suchas polyamines, small salts, ATP and RNA, cause on/off switching in the higher-order structure of DNA. Thus, the field parameters of the cellular environment should play important roles in the mechanism of self-regulation, in addition to networks of key and locks. This conformational transition induced by field parameters may be related to rigid on/off regulation, whereas key-lock relationships may be involved in a more flexible control of gene expression.