Enhanced surface plasmon resonance immunosensing using a streptavidin-biotinylated protein complex

In this paper, we present a novel strategy for improving the sensitivity of surface plasmon resonance immunosensing using a streptavidin-biotinylated protein complex. This amplification strategy is based on the construction of a molecular complex between streptavidin and biotin labeled protein. The complex can be formed in a cross-linking network of molecules so that the amplification of the response signal will be realized due to the big molecular size of the complex. The results show that the amplification strategy causes a dramatic improvement of the detection sensitivity. hIgG protein could be detected in the range of 0.005-10 mug ml(-1).

Determination of the impurities elements in high-purity cadmium by inductively coupled plasma mass spectrometry

A method for the determination of impurity elements in high purity cadmium by inductively coupled plasma mass spectrometry was developed. The spectral interference arising from Cd was discussed and the magnitude of interference by the polyatomic ion of Cd was compared. The effect of the high concentration Cd on the analyte signal was studied. The results showed that Cd possessed both significant suppression effects on the light mass elements signal and enhancement effects on the heavy mass elements signal,and the effects can be corrected by the use of internal standard elements. In this paper, Y and Bi was employed for the light mass elements and the heavy mass elements,respectively. As, Be, Co, Cu, Ga, Ge, Mn, Mo, Pb, Ni, Sr, Au, Tl, Th, V and U in high purity cadmium were determined. Detection limits are 0.005-0.052 mu g L-1,and the recoveries of standard addition are 82%-108%.

Oxidation of NADH by dopamine incorporated in lipid film cast onto a glassy carbon electrode

Stable lipid film was made by casting lipid in chloroform onto a glassy carbon electrode. This model of a biological membrane was used to investigate the oxidation of dihydronicotinamide adenine dinucleotide (NADH) by dopamine. After this electrode had been immersed in dopamine solution for 10 h, it was found that some dopamine had been incorporated in the film. The cyclic voltammogram was obtained for the oxidation of 2.0 X 10(-3) mol 1(-1) NADH with dopamine incorporated in the films. All electrochemical experiments were performed in 0.005 mol 1(-1) phosphate buffer (pH 7.0) containing 0.1 mol 1(-1) NaCl without oxygen. The oxidation current increased gradually with successive sweeps and reached steady state. It was a different phenomenon from previous results. The anodic overpotential was reduced by about 130 mV compared with that obtained at a bare glassy carbon electrode. The diffusion coefficient for 2.0 X 10(-3) mol 1(-1) NADH was 6.7 X 10(-6) cm(2) s(-1). (C) 1999 Elsevier Science S.A. All rights reserved.

Analysis of mussel standard reference material by inductively coupled plasma atomic emission spectrometry and inductively coupled plasma mass spectrometry

A method for the analysis of mussel standard reference material by inductively coupled plasma atomic emission spectrometry( ICP-AES) and inductively coupled plasma mass spectrometry(ICP-MS) was developed. K, Na, Ca, Mg, P, Al, Fe, Zn, Mn and Sr were determined by ICP-AES and As, B, Cd, Co, Cr, Cu, Ga, Ge, Mn, Mo, Ni, Pb, Se, Sr, U and V by ICP-MS, The interference coefficients at the Mn-55, Se-78, Cu-63, Co-59, Ni-58, Ni-60, As-75, Se-77, V-51, Cr-53 and Cr-52 originating from polyatomic ion of the matrix elements (KO)-K-39-O-16, K-39(2), (ArNa)-Ar-40-Na-23, (CaO)-Ca-43-O-16, (CaO)-Ca-42-O-16, (CaO)-Ca-44-O-16, (PO2)-P-31-O-16, (ArCl)-Ar-40-Cl-35, (ArCl)-Ar-40-Cl-37, (ClO)-Cl-35-O-16, (ClO)-Cl-37-O-16 and (ArC)-Ar-40-C-12 were determined under the selected operation parameters. The major matrix elements, such as K, Na and Ca, result in the suppression of analytes signals. The apparent concentration at the significant biological element which was produced by the different digestion methods, (.) HNO3 + H2O2 (3 + 2), HNO3 + HClO4 (3 + 0.5) and HNO3 + H2SO4 (3 + 0.5),was determined. The sample digested by HNO3 + H2O2 did not give rise to interfere on the analyte, and the backgrounds of Se-77, Ga-69, Zn-67, As-75, V-51, Cr-53 and Cr-52 were increased by HNO3 + HClO4 digestion method, that affected the determination of these elements, especially the monoisotope As and V. Sample digested by HNO3 + H2SO4 increased the backgrounds at Cu-65, Zn-64 and Zn-67. Detection limits of ICP-AES are 0.001 similar to 0.75 mg/L and those of ICP-MS are 0.005 similar to 1.01 mu g/L. The relative standard derivations of ICP-AES and ICP-MS are 2.7% similar to 12.8%, 3.4% similar to 24.8%, respectively.

电感耦合等离子体质谱法测定高纯镉中的杂质元素

采用电感耦合等离子体质谱法测定高纯镉中的杂质元素。讨论了镉基体的谱线干扰，比较了镉基体产生的多原子离子的干扰程度，该干扰按Cdh、CdO、CdO2、CdAr、CdOH和CdOH2顺序递减。研究了不同浓度镉基体对分析物信号的抑制或增强效应，镉的浓度大于0.25·L-1时，对质量数小于100的分析物的信号强度产生抑制，而对质量数大于150的分析物的信号强度产生增强，采用89Y和209Bi作内标分别克服基体的抑制和增强效应。测定了高纯镉的As、Be、Co、Cu、Ga、Ge、Mn、Mo、Pb、Ni、Sr、Au、Tl、Th、V和U等16个杂质元素，方法的检测限0.005～0.052μｇ·L-1，标准加入回收率82%～108%。

Synthesis and crystal structure of aluminium complex with 3-hydroxyflavone

[Al(C15H9O3)(3)](2) . 2CHCl(3) . 8H(2)O was synthesized, and its crystal structure was determined. It belongs to trigonal system, R3, a=b=1. 655 8(3) nm, c=3. 646 5(20) nm, alpha = beta = 90 degrees, gamma = 120 degrees, V = 8. 656 08(0. 005 86) nm(3). D-c = 1.45 g/cm(3), mu(Mo K alpha) = 3. 20 cm(-1), F (000) = 3 924. The crystal structure was solved by Patterson and Fourier techniques, and refined by a block-diagonal least-squares method. A total of 3 737 independent intensity data were collected, of which 1 033 with I greater than or equal to 3 sigma(I-0) were observed, R = 0. 091 8, Rw=0. 091 8. Al3+ ion was 6-coordinated, bound to six oxygen atoms from three 3-hydroxyflavones to form a distortional coordination octahedron.

THE APPLICATION OF THE MPB4 THEORY TO THE INTERFACE BETWEEN 2 IMMISCIBLE ELECTROLYTE-SOLUTIONS .2. THE DIFFERENTIAL CAPACITANCE OF THE WATER 1,2-DICHLOROETHANE INTERFACE

The MPB4 theory is used to calculate the differential capacitance of the interface between LiCl in water and TBATPB in 1,2-dichloroethane at electrolyte concentrations of 0.005, 0.01 and 0.02 M. The effects of the ion size and the image force, and the influence of the electrolyte concentration, the surface charge density and the solvent effect on the inner layer potential drop are considered simultaneously. These effects can be ascribed to the ionic penetration into the opposite solution and ion-ion correlations across the interface. Our results are in better agreement with experimental data than those obtained using Gouy-Chapman theory. This indicates that the MPB4 theory may also describe the structure of the water \1,2-dichloroethane interface provided that the influence of the electrolyte concentration, the surface charge density and the solvent effect on the inner layer potential distribution are included in the calculation. Comparison of the theoretical results with those of the water \nitrobenzene interface shows that the structure of the water \1,2-dichloroethane interface is similar to that of the water \nitrobenzene interface, except that in the former case the inner-layer potential drop is much higher and the effects of the image force and the ion size are more pronounced.

ISOPOLYMOLYBDIC ACID-POLY(4-VINYL)PYRIDINE FILM MODIFIED ELECTRODE

Electrochemical polymerization of 4-vinylpyridine produced a uniform poly(4-vinyl)pyridine(PVP) film on the glassy carbon (GC) electrode surface. The isopolymolybdic acid-PVP film-modified electrode was prepared by soaking the PVP/GC electrode in the 0.05 M H2SO4 aqueous solution containing 0.005 M isopolymolybdic acid (H4Mo8O26). The latter (catalyst) is incorporated and held in the PVP film electrostatically. The electrochemical behavior and electrocatalytic properties of this H4Mo8O26-PVP/GC electrode was described. The results indicate that this modified electrode has good stability and electrocatalytic activity on the reduction of chlorate and bromate ions in aqueous solution. The catalytic process is regarded as an EC mechanism.

Ce_(0.95)Ca_(0.05)F_(3-x)固体电解质氧传感器的研究

本文研究了Ce_(0.95)Ca_(0.05)F_(3-x)固体电解质及其氧传感器的制备和性质。在150℃时,Po_2在10~(-2)～10~(-1)MPa范围内传感器件的电位变化正比于Po_2的对数值。同时研究了不同参比电极和敏感电极对传感性能的影响。以Bi+BiF_3为参比电极时,EMF值稳定较快;当Pd作为敏感电极时,传感元件的响应较快;以RuO_2作敏感电极时,传感器件较灵敏。文章还讨论了敏感机理。

Development of expressed sequence tags from the bay scallop, Argopecten irradians irradians

The bay scallop, Argopecten irradians irradians, introduced from North America, has become one of the most important aquaculture species in China. Inan effort to identify scallop genes involved in host defense, a high-quality cDNA library was constructed from whole body tissues of the bay scallop. A total of 5828 successful sequencing reactions yielded 4995 expressed sequence tags (ESTs) longer than 100 bp. Cluster and assembly analyses of the ESTs identified 637 contigs (consisting of 2853 sequences) and 2142 singletons, totaling 2779 unique sequences. Basic Local Alignment Search Tool (BLAST) analysis showed that the majority (73%) of the unique sequences had no significant homology (E-value >= 0.005) to sequences in GenBank. Among the 748 sequences with significant GenBank matches, 160 (21.4%) were for genes related to metabolism, 131 (17.5%) for cell/organism defense, 124 (16.6%) for gene/protein expression, 83 (11.1%) for cell structure/motility, 70 (9.4%) for cell signaling/communication, 17 (2.3%) for cell division, and 163 (21.8%) matched to genes of unknown functions. The list of host-defense genes included many genes with known and important roles in innate defense such as lectins, defensins, proteases, protease inhibitors, heat shock proteins, antioxidants, and Toll-like receptors. The study provides a significant number of ESTs for gene discovery and candidate genes for studying host defense in scallops and other molluscs.

四角蛤蜊Mactra veneriformis对镉和汞污染胁迫的生理响应

本论文基于对我国沿海重金属污染加剧以及滩涂贝类资源衰退现象的关注，围绕典型滩涂贝类四角蛤蜊生物化学、细胞生物学和免疫学特征开展研究，调查了渤海湾天津近岸海域四角蛤蜊体内重金属含量和分布规律，以及四角蛤蜊对沉积物重金属的富集能力。室内模拟研究了四角蛤蜊对重金属镉和汞的富集能力及同化机制（assimilation mechanism）差异性，四角蛤蜊各组织对镉和汞的解毒机理和负载能力，以及重金属胁迫对四角蛤蜊血细胞结构和功能的损伤效应。研究结果可望为探讨四角蛤蜊受重金属胁迫细胞水平上的响应机制，揭示重金属污染和贝类资源衰退的关系，建立快速可靠的重金属污染生物标志物指标体系，开展生态健康评估和生境修复等提供科学依据。主要研究结果如下： 1．查明了渤海湾天津近岸四角蛤蜊体内和表层沉积物中Cd、Pb、Cu、Zn、Mn、Cr和Ni七种重金属的含量和周年时空分布特征；表层沉积物中Cd具有较强的污染程度和潜在生态风险，四角蛤蜊对重金属元素Cd和Pb具有较高的富集能力，对环境重金属污染具有很好的指示作用。 2．查明了Cd对四角蛤蜊在24h，48h和96h的半致死浓度（LC50）分别为15.96mg/L、5.15mg/L和2.38mg/L，汞的24h，48h和96h LC50分别为3.71mg/L，0.61mg/L和0.21mg/L。镉和汞对四角蛤蜊的安全质量浓度分别为0.0238mg/L和0.0021mg/L。 3．四角蛤蜊软体部对Cd和Hg的富集能力有显著差异。暴露过程中，四角蛤蜊软体部Cd和Hg的增加量分别为0.12-7.7µg/g和0.002-0.024µg/g，富集率分别为0.3-6.2%和0.11-0.68%，吸收率常数分别为0.07-1.10和0.001-0.005。 4．四角蛤蜊受Cd和Hg胁迫后，外套膜、鳃和肝胰腺的金属硫蛋白（MT）含量均在暴露浓度和胁迫时间上都有极显著变化。组织内MT含量的大小关系为：肝胰腺＞鳃＞外套膜。肝胰腺可以作为双壳贝类MT重金属污染指示研究的目标组织。四角蛤蜊对Cd和Hg不同的解毒机制，导致组织中MT的表达含量和Cd胁迫显著相关，而与Hg胁迫无显著相关性。 5．四角蛤蜊肝胰腺和鳃中抗氧化系统酶、脂质过氧化产物、酸性磷酸酶（ACP）和碱性磷酸酶（ALP）在不同Cd胁迫浓度和暴露时间下有显著差异。与对照组相比，超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPx）的活力和丙二醛（MDA）含量均随着暴露时间的增加而增加，达到一个峰值，然后减低。而ACP和ALP活力则表现出先被抑制，然后升高的变化趋势。肝胰腺中CAT和GPx活力高于鳃，鳃中SOD和MDA含量高于肝胰腺，这种差异与两种组织不同的解毒机理有关。 6．重金属胁迫能够造成四角蛤蜊血细胞超微结构的损伤、溶酶体膜稳定性改变、微核和总畸形核生成，各种损伤均表现出明显的浓度依赖效应。2μg/L Hg暴露14天后，血细胞溶酶体膜中性红保持时间（NRR）、微核生成率（MN）和总畸形核生成率（TNA）与对照组相比没有显著差异。相同暴露时间下，25 μg/L Hg处理组血细胞NRR值低于25 μg/L Cd处理组，而MN和TNA值则相反，说明Hg胁迫对四角蛤蜊血细胞溶酶体膜具有较强的损伤作用。研究发现，NRR、MN和TNA三种指标对于衡量重金属污染对四角蛤蜊血细胞的毒性效应有很好的协同检测作用，可望作为有效的生物标志物在环境监测技术中得以应用。

栉孔扇贝抗鳗弧菌感染性状候选基因的多态性研究

栉孔扇贝（Chlamys farreri）是我国北方地区主要的养殖贝类之一，曾为沿海各省带来巨大的经济效益。但自1997年以来，陆续爆发的病害问题给扇贝养殖业造成了巨大的经济损失，严重影响了该产业的健康发展。目前认为培育抗病性强的扇贝优良品种是解决病害问题的根本途径。由于传统的育种方法费时费力，无法满足对良种的迫切需求，因此有必要通过分子手段加快抗病品种的培育步伐。标记辅助育种（marker assisted selection，MAS）是成功应用于动物育种中的分子手段之一，但由于缺乏与抗病性状相关的标记，MAS目前还无法在软体动物中得到应用。因此，寻找与抗病性状相关的分子标记是在软体动物中发展MAS的关键。 本研究利用鳗弧菌（Listonella anguillarum）对栉孔扇贝进行攻毒感染实验，初步得到敏感群体和抗病群体后采用PCR、PCR-RFLP、Bi-PASA PCR等方法研究了CfLysG、CfC1qDC和CfLITAF基因多态性及其与栉孔扇贝对鳗弧菌抗性的关系。 研究发现，栉孔扇贝CfLysG的基因序列中共有104个单核苷酸多态性（SNP）位点和29个插入/缺失（I/D）多态性位点。有17个多态性位点位于启动子区域，选择其中的-753 I/D、-391A/G和-284I/D多态性进行检测，发现这三个位点的基因型在敏感群体和抗病群体中的分布均符合Hardy-Weinberg平衡（P>0.05）。其中-753 ID基因型和-284 ID基因在抗病群体中的频率高于在敏感群体中的频率，但两者之间无显著性差异（P>0.05）。-391 AG基因型在抗病群体中的频率显著高于敏感群体（P=0.007），表明-391 AG基因型与栉孔扇贝对鳗弧菌的抗性显著相关。为验证这一相关性，对-391位点不同基因型的扇贝进行攻毒感染实验。统计发现，具有-391 AA基因型的扇贝累计死亡率显著高于具有-391 AG基因型的扇贝（P=0.001），进一步证实了CfLysG基因-391 AG基因型与栉孔扇贝对鳗弧菌的抗性显著相关。CfLysG基因的外显子共有3处SNP，其中仅第三外显子上的+3473 A/C为非同义突变。统计分析表明，+3473位点不同基因型在敏感群体中的分布频率符合Hardy-Weinberg平衡（P>0.05），而在抗病群体中则偏离Hardy-Weinberg平衡（P<0.01）。+3473 AA基因型在抗病群体中的频率显著高于在敏感群体中的频率（P=0.022），表明+3473 AA基因型与栉孔扇贝对鳗弧菌的抗性显著相关。CfLysG基因第1内含子存在+96 I/D和+487 I/D两处大片段的I/D多态性。统计发现，这两个位点的基因型在敏感群体和抗病群体中的分布频率均符合Hardy-Weinberg 平衡（P>0.05）。其中+96 DD基因型和+487 ID基因型在抗病群体中的频率均略高于在敏感群体中的频率，但两者之间无显著性差异（P>0.05）。表明这两个位点的多态性与栉孔扇贝对鳗弧菌的抗性无显著相关性。对CfLysG基因各多态性位点的统计分析表明，各位点之间存在不同程度的连锁不平衡，提示有单体型的存在。对19种频率>1%的单体型在敏感群体及抗病群体中的频率进行分析，发现-753 I/-391 G/-284 I/+96 I/+487 D/+3473 A单体型在抗病群体中的频率显著高于敏感群体（P=0.044），表明该单体型与栉孔扇贝对鳗弧菌的抗性显著相关。 在栉孔扇贝CfC1qDC基因cDNA序列上共发现14处SNP。对+423 T/C多态性与栉孔扇贝对鳗弧菌抗性的关系进行了分析。统计发现，+423位点各基因型在敏感群体和抗病群体中的分布均符合Hardy-Weinberg平衡（P>0.05）。+423 TT基因型在抗病群体中的频率显著高于在敏感群体中的频率（P=0.005），表明+423 TT基因型与栉孔扇贝对鳗弧菌的抗性显著相关。 在栉孔扇贝CfLITAF基因cDNA序列中共发现3处SNP及1处I/D多态性。对+145 I/D多态性进行研究，发现所有敏感个体及抗病个体中均同时存在+145 位点所有等位基因，表明+145位点多态性与栉孔扇贝对鳗弧菌的抗性不相关。 以上研究表明，栉孔扇贝CfLysG基因-391 AG基因型、+3473 AA基因型、-753 I/-391 G/-284 I/+96 I/+487 D/+3473 A单体型以及CfC1qDC基因+423 TT基因型与栉孔扇贝对鳗弧菌的抗性显著相关，提示它们可作为与栉孔扇贝抗病相关的候选分子标记应用于贝类抗病育种中，为贝类的标记辅助育种提供参考。此外，抗病相关分子标记的发现还有利于加深对扇贝发病机理的理解，并有助于发掘预防及治疗贝类疾病的新方法。

Impact of microzooplankton and Calanus sinicus (Brodsky, 1962) on phytoplankton in the Yellow Sea during early summer

Dilution incubations and Calanus sinicus addition incubations were simultaneously conducted at five stations in the Yellow Sea in June of 2004 to evaluate the impact of microzooplankton and Calanus sinicus on phytoplankton based on the Chlorophyll a (Chl-a) levels. The Chl-a growth rates (k) ranged from 0.60-1.67 d(-1), while microzooplankton grazed the Chl-a at rates (g) of 0.29-0.62 d(t-1). The addition of C. sinicus enhanced the Chl-a growth rate (Z) by 0.004-0.037 d(-1) ind.(-1) L. C. sinicus abundance ranged from 84.1-160.9 ind. m(-3), which occupied 90.7%-99.1% of the copepod (> 500 mu m) population. The in-situ increase in phytoplankton by C. sinicus community was estimated to be 0.000 4-0.005 9 d(-1). These results showed that microzooplankton were the main grazers of phytoplankton, while C. sinicus induced a slight increase in the levels of phytoplankton.

碱性蛋白酶降解骨胶原制备食品明胶工艺

[目的]研究碱性蛋白酶降解骨胶原的工艺条件,探索其应用于骨料制备明胶的可行性.[方法]采用碱性蛋白酶降解骨胶原制备食品明胶,通过正交试验L_9（3~4）得出酶解制备食品明胶的最佳工艺参数.[结果]结果表明：最佳工艺参数为20万U/g碱性蛋白酶用量为干骨质量的0.005%,酶解温度60℃,酶解pH值为8,酶解时问5 h,升温灭酶,提胶一次2 h;获得食品明胶指标值分别为：得胶率20.0%,黏度3.1 mPa•s,冻力193 Bloomg.[结论]该研究为获得品质和产量俱佳的食品明胶提供了试验依据.

青藏高原高山植物麻花艽的传粉生态学研究

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