呋喃唑酮人工抗原的制备与鉴定

为了制备3-氨基-2-恶唑烷酮(AOZ)的完全抗原,作者以乙醇肼和碳酸二甲酯为原料,首先合成了3-氨基-2-恶唑烷酮(AOZ),然后通过对醛基苯甲酸对AOZ进行衍生制得3-(4-羧基苯亚甲基)-氨基-2-恶唑烷酮(CPAOZ),以CPAOZ作为半抗原,经水溶性碳化二亚胺法(EDC),缩合酯化法(DCC)和混合酸酐法(MA)法3种方法分别与载体蛋白偶联,合成人工抗原。对于制备产物通过紫外光谱扫描分析进行了鉴定,3种抗原的偶联比率分别为8、12、17。采用3种抗原做为包被原,建立了酶联免疫(ELISA)抑制曲线。结果表明,对AOZ的半数抑制率(IC50)分别为12.9、7.6、1.1 ng/mL。采用混合酸酐法(MA法)制备的包被抗原与抗体具有较好的匹配性。3-氨基-2-恶唑烷酮(AOZ),是硝基呋喃类抗生素呋喃唑酮在动物体内的稳定代谢物,采用本试验制备的抗原,有望获得特异性更好的抗体。     

基于不同半抗原的呋喃唑酮代谢物免疫检测方法的建立与比较

为实现呋喃唑酮代谢物3-氨基-2-恶唑烷酮（3-amino-2-oxazolidinone,AOZ）的快速定量检测,制备AOZ- 牛血清蛋白单克隆抗体,建立2 种分别基于抗3-（4-羧基苯亚甲基）-氨基-2-恶唑烷酮（CPAOZ）、抗AOZ的单克 隆抗体酶联免疫检测试剂盒。CPAOZ检测试剂盒在1.0～100.0 ng/mL范围具有较好的线性,IC50值为14.6 ng/mL, 检测限为6.56 ng/mL,回收率为97.6%～101.3%;AOZ检测试剂盒在0.5～12.5 ng/mL范围具有良好线性,IC50值为 3.9 ng/mL,检测限为0.45 ng/mL,回收率为94.1%～97.4%。这2 种检测试剂盒对于其他硝基呋喃类抗生素及其代谢 物均不存在交叉反应。从经济、方便、快速、灵敏等因素来考虑,后者更具有发展前景。     

呋喃唑酮残留标示物人工抗原合成与抗体制备

以乙醇肼和碳酸二甲酯为原料,合成2种半抗原3-氨基-2-恶唑烷酮(AOZ)和3-(4-羧基苯亚甲基)-氨基-2-恶唑烷酮(CPAOZ),经戊二醛法和碳二亚胺法将半抗原与牛血清蛋白(BSA)偶联合成2种人工抗原AOZ-BSA和CPAOZ-BSA,产物经紫外光谱扫描分析鉴定。用2种人工抗原采取不同程序免疫新西兰大白兔,获得8组多克隆抗体。AOZ-BSA抗体效价1:0.8×104～1:3.2×104,对AOZ灵敏度低,IC50均大于100μg/ml。CPAOZ-BSA抗体效价1:1.6×105～1:6.4×105,对AOZ与苯甲醛衍生物N-苯亚甲基-3-氨基-2-恶唑烷酮(PAOZ)灵敏度高,IC500.91～11.56ng/ml,特异性好,仅与呋喃唑酮(交叉反应率6.15%)、3-(2-硝基苯亚甲基)-氨基-2-恶唑烷酮(NPAOZ,交叉反应率5.73%)有交叉反应,与衍生试剂苯甲醛和其他药物没有交叉反应(交叉反应率<0.01%)。本研究制备出AOZ特异性抗体,为我国开发AOZ酶联免疫试剂盒奠定基础。     

呋喃唑酮代谢物间接竞争化学发光酶免疫法的建立

建立快速检测动物源性食品中呋喃唑酮代谢物AOZ的间接竞争化学发光酶免疫法。利用对醛基苯甲酸(4-CBA)将AOZ衍生化为CPAOZ,利用活化酯法将CPAOZ与卵清蛋白(OVA)偶联,生成完全抗原CPAOZ-OVA。对包被原与抗体的最佳稀释倍数、封闭液浓度、竞争时间、酶标二抗稀释倍数进行优化,建立标准曲线,同时对方法的特异性进行评价。结果表明:包被抗原最佳稀释倍数为2000倍,抗体的最佳稀释倍数为20000倍、封闭液为2%脱脂乳粉、竞争时间为2 h、二抗最佳稀释倍数为4000倍;线性范围是0.075~7.396 ng/m L,半抑制浓度IC_(50)为0.74 ng/m L,对呋喃唑酮原药的交叉反应率为23.4%,与其他结构类似物及衍生化试剂的交叉反应率均小于0.1%。该方法灵敏度高、特异性好,可为监管部门对于快速检测动物源性食品中AOZ提供依据。     

抗呋喃唑酮代谢物单链抗体基因构建及蛋白结构分析和活性鉴定

从单克隆杂交瘤细胞出发,构建抗3-氨基-2-噁唑烷酮（3-amino-2-oxazolidinone,AOZ）衍生物单链抗体基因,并对其蛋白质进行理化性质分析及结构预测和活性鉴定。首先以呋喃唑酮代谢物AOZ衍生物单克隆抗体杂交瘤细胞1D2为原料,提取总RNA后克隆重链（VH）、轻链（VL）可变区基因,然后使用重叠延伸聚合酶链式反应技术用连接肽(G4S)3将VH、VL基因连接构建抗AOZ衍生物单链抗体基因,经测序后采用生物信息软件对抗体编码的氨基酸序列进行推导并展开生物信息学分析,再将此基因与Plip6/GN表达载体连接后进行表达并采用直接竞争酶联免疫分析法（direct competitive enzyme-linked immunosorbent assay,dcELISA）鉴定其活性。结果表明：抗AOZ衍生物单链抗体基因全长750?bp,编码250?个氨基酸,相对分子质量为27?012.20,理论等电点为9.13,属于碱性氨基酸;其二级结构预测结果显示抗体蛋白含20?处α-螺旋、25?处β-转角、114?处无规卷曲、91?处延伸带结构;三级结构预测的正面俯视图显示重、轻链由连接肽相连形成一个“环桶状”结构,侧面显示为“口袋状”,这与常规单链抗体的结构特征一致,dcELISA结果也显示该抗体具有良好的抗原结合活性。本研究为后续AOZ残留检测免疫分析方法的建立及抗AOZ衍生物单链抗体的改造奠定一定基础。     

基于衍生化3-氨基-2-恶唑烷酮的夹心酶联免疫分析

建立一种3-氨基-2-恶唑烷酮（3-amino-2-oxazolidinone,AOZ）夹心免疫检测方法。通过1,6-己二醇连接2-硝基-4-羧基苯甲醛和生物素合成针对AOZ的新型衍生试剂。在样品前处理时加入衍生试剂可对AOZ进行衍生,衍生产率为89%。在酶标板上包被抗AOZ单克隆抗体并以辣根过氧化物酶标记的亲和素或抗生物素抗体作为第二结合物可实现AOZ的夹心酶联免疫吸附检测（enzyme-linked immunosorbnent assay,ELISA）。从实际应用的角度,得到双抗夹心和抗体-亲和素夹心模式下两个表位的极限距离,分别为12?和13?,理想距离分别为16?和17?。在双抗夹心和抗体-亲和素夹心模式下的检出限分别达到1.8 pg/mL和0.8 pg/mL（以AOZ质量浓度计）,相对于竞争ELISA,其灵敏度最高提高了25倍。平均回收率为73%～85%,平均相对标准偏差为9.0%。     

酶联技术在水产品中呋喃唑酮代谢物AOZ的检测中的应用

用酶联免疫检验技术（ELISA）检测呋喃唑酮（Furazolidone）代谢物AOZ。AOZ经过16h的衍生后和酶联结合物对微孔中的抗体进行竞争反应，利用抗原与抗体的特异性免疫化学反应的基本原理，在酶标仪上450nm处测定吸光强度。进行定性定量分析。     

不同抗生素参与下呋喃唑酮代谢物 在鲫鱼体内的残留消除规律

本文研究了土霉素、恩诺沙星、磺胺间甲氧嘧啶分别与呋喃唑酮联合药浴给药后,呋喃唑酮代谢物3-氨基-2-恶唑烷酮(AOZ)在鲫鱼体内的残留消除规律。结果表明,呋喃唑酮在进入鲫鱼体内后迅速代谢成AOZ,停药0 h,呋喃唑酮对照组中鲫鱼肾脏、肝脏、肌肉中AOZ残留量便达到最高,分别为88.87、59.20、14.91μg/kg,平均消除速率为0.86、0.52、0.14μg/(kg·h);与对照组相比,土霉素-呋喃唑酮组鲫鱼肾脏、肝脏、肌肉中AOZ最大残留浓度明显增加,分别为165.40、125.29、28.69μg/kg,平均消除速率为1.71、1.17、0.31μg/(kg·h);而恩诺沙星-呋喃唑酮组鲫鱼肾脏、肝脏、肌肉中AOZ最大残留浓度与对照组相比显著减少,仅为46.70、41.64、9.69μg/kg,平均消除速率为0.36、0.31、0.03μg/(kg·h);磺胺间甲氧嘧啶-呋喃唑酮组中鲫鱼体内AOZ最大残留量及平均消除速率与对照组相比也有差异但变化不大。消除96 h后,各实验组鲫鱼体内仍有大量AOZ残留。本实验初步表明AOZ在鲫鱼体内难消除,且土霉素、恩诺沙星、磺胺间甲氧嘧啶能够不同程度的影响AOZ在鲫鱼体内的残留与消除,这为水产品质量安全监管和渔药残留研究提供了理论基础。     

间接竞争ELISA法检测呋喃唑酮代谢物

以人工抗原和特异性抗体为基础,建立了以2-硝基苯甲醛为衍生试剂检测呋喃唑酮代谢物3-氨基-2-恶唑烷酮(AOZ)间接竞争ELISA方法。通过方阵滴定和间接竞争法确定ELISA方法的抗原抗体的最佳工作浓度;得到标准曲线的线性范围为0.25~10ng/mL,线性关系良好;根据标准曲线计算IC50为0.5881~1.1333 ng/mL。此外,检测了与其他类似物的交叉反应率,显示了很好的特异性;并且通过对虾样品添加回收率的测定,证明了该方法的准确性。高温短时间衍生化处理可达到与37℃孵育过夜同样的衍生效果。     

化学发光酶免疫法测定动物源性食品中呋喃唑酮代谢物的含量

目的建立化学发光酶免疫法(chemiluminescent enzyme immunoassay, CLEIA)测定动物源食品中呋喃唑酮代谢物的分析方法。方法通过使用N-羟基琥珀酰亚胺酯法将衍生的CPAOZ半抗原与卵白蛋白(ovalbumin, OVA)缀合以合成包被抗原,优化CLEIA酶免疫方法的主要条件。对CLEIA酶免疫法的灵敏度、特异性、精密度及准确度进行评价,并就准确度指标同国家标准中的液相色谱串联质谱法进行对比分析。结果包被原和单克隆抗体稀释倍数分别为4000倍和1600倍, 1%脱脂乳封闭,竞争时间30 min, HRP-IgG孵育时间60 min时为最佳条件。CLEIA的线性方程为Y=-0.357X+1.459(r~2=0.984), IC50为0.486 ng/mL,线性范围为0.070～3.362 ng/mL。回收率在87.9%～92.6%之间,批内和批间变异系数分别为3.6%～7.5%和3.2%～6.3%。结论该方法简单,快速,可用于实验室或现场动物源性食品的AOZ筛选。     

蛋白质水解度测定方法综述

对目前国内外常用的蛋白质水解度测定方法进行了综述，其中pH—state方法是通过滴定水解过程中释放的质子测定DH；OPA、TNBS及国内常用的水合茚三酮和甲醛等测定方法是利用游离氨基的反应测定DH。     

SEROLOGIC IDENTIFICATION OF SPECIES OF ORIGIN OF SAUSAGE MEATS

SUMMARY: A partially purified immunoglobulin G (lgG) solution prepared from the serum of species to be tested was heated to the specifications for sausages. The resulting supernatant fluid was decanted and the precipitate washed with saline and used to immunize rabbits. The supernatant fluid was used to sensitize tanned sheep red blood cells. The immune serum was rendered monospecific by absorptions with heterologous, heated lgG precipitates. A sample of monospecific immune serum was absorbed with a washed homogenate of sausage. Aliquots of the monospecific immune serum, both untreated and sausage absorbed, were tested with cells sensitized with the homologous heated lgG supematant fluid. A significant reduction of titer by sausage absorption indicated that the sausages contained the meat homologous to the immune serum.     

BASIS OF STABILITY OF AMINE SALTS OF LINOLEIC ACID

SUMMARY— The mechanism and generality of the known stabilization against autoxidation conferred on linoleic acid by certain basic amino acids, such as lysine and arginine, was investigated. Basic amino acids were the only class of compounds found to confer the effect. However, the smallest basic amino acid, 2,3-diaminopropionic acid, was not effective, nor was αβω-diaminc acid, 3,6-diaminohexanoic acid, although a simple isomer of lysine. The stabilization was observed only in the solid phase. Inclusion of sodium chloride in the solid matrix was deleterious to the effect. A large number of physical and chemical observations were made and correlated but it has not been possible to draw detailed conclusions about the mechanism of stabilization, nor can a detailed structure of the stabilized complex be suggested. The cause of the phenomenon appears to be closely associated with the physical arrangement of the ions in the crystal lattice.     

聚多元羧酸盐的合成及应用

研究了聚多元羧酸盐的合成方法及反应机理,将其应用于洗涤剂和PVC制品中分别代替三聚磷酸钠和邻苯二甲酸二丁酯,证明有良好效果。     

百年风尚

一场流光溢彩、赏心悦目的展览,一段百年风尚演进的传奇旅程,一次东西方文化艺术的完美对话。2013年9月13日,“博萃臻艺一中西方珍宝艺术展”在辽宁省博物馆举行了隆重的开幕仪式,法兰西共和国驻华大使白林女士、辽宁省文物局局长丁辉先生、辽宁省博物馆馆长马宝杰先生、卡地亚全球总裁兼首席执行官邓阁仕先生、卡地亚区域行政总裁（北亚洲）陆慧全先生、卡地亚中国区首席执行官陆意斯先生、辽宁省文物店总经理张春鹰先生,以及众多文化界与文博界的贵宾齐聚一堂,共同见证了这场文化艺术盛事。     

STABILITY OF AQUEOUS EMULSIONS OF THE ESSENTIAL OIL OF HOPS

Hop oil emulsions prepared from different varieties of hops have been found to exhibit enhanced physical stability on the addition of blends of the emulsifiers Span 20/Tween 80 or Span 60/Tween 60. Examination of the particle size and volume distributions of an emulsion by use of a Coulter Counter was found to be an excellent method of monitoring its stability. An indication as to the relative efficiency of emulsifiers can be obtained from Coulter Counter measurements on hop oil emulsions after storage for 4 days. The use of an ultracentrifuge provies a rapid means of testing emulsion stability and hence the effectiveness of emulsion stabilizers.     

EFFECTS OF TYPES OF FAT AND OF RATES AND TEMPERATURES OF COMMINUTION ON DISPERSION OF LIPIDS IN FRANKFURTERS

ABSTRACT— The effect of time, temperature and rpm of comminution of emulsions was determined on the dispersion of approximately 25% of beef fat, pork fat or cottonseed oil in frankfurters. The numbers of lipid particles 5 μ or less in diameter increased in frankfurters containing either beef or pork fat as comminution was continued to higher temperatures, with pork fat dispersed more thoroughly. Fat tended to separate from frankfurters containing beef fat in particles 200 μ or more in diameter. In contrast, no specific degree of dispersion of particles 5 μ or less in diameter consistently indicated emulsion stability, or its lack. Increased rpm during comminution produced an increased dispersion of beef or pork fat. Under the same conditions pork fat was dispersed more finely than beef fat. Dispersion of cottonseed oil produced finely dispersed particles beyond the resolution of light microscopy, as was confirmed by electron microscopy which showed a substantial number of particles to be less than 1 μ in diameter.     

DEVELOPMENT OF CHAINS OF CELLS OF SACCHAROMYCES CEREVISIAE DURING FERMENTATION

The lengths of chains of cells of Saccharomyces cerevisiae were studied during fermentation. Pitching yeast generally contained about half of the total number of cells as two-celled chains. The chain lengths varied during the subsequent fermentation and the variations were characteristic of the strain. Electronic counting assessments of chain length were unreliable.     

ANALYTICAL CHARACTERIZATION OF THE PRODUCTS OF NONENZYMATIC GLYCOSYLATION OF LYSOZYME

The quantitative analysis of the reaction products of the water activity dependent nonenzymatic glycosylation of lysozyme was not straightforward. Difficulties arose in the determination of the number of bound glucose molecules because glycosylation leads to glucose mediated protein aggregation, and the likely presence of a mixture of relatively labile Schiff-base intermediates, and the more stable ketoamine products generated by Amadori rearrangement. Polyacrylamide gel electrophoresis was used to monitor protein aggregation; periodate oxidation, nuclear magnetic resonance (NMR), and oxalic acid hydrolysis combined with HPLC, emerged as the most promising methods to quantitate the degree of glycosylation. Possible interpretations are advanced to explain the apparent discrepancies in degree of glycosylation suggested by the different analytical methods evaluated.     

矩阵乘积的行式,列式

给出了ｍ×ｍ矩阵与ｍ×ｎ矩阵的行（列）式的表达式．若Ａ＝ａ１１ａ１２…ａ１ｍａ２１ａ２２…ａ２ｍ……ａｍ１ａｍ２…ａｍｍＢ＝ｂ１１ｂ１２…ｂ１ｎｂ２１ｂ２２…ｂ２ｎ……ｂｍ１ｂｍ２…ｂｍｎ分别是ｍ×ｍ，ｍ×ｎ矩阵，则｜Ａ｜｜Ｂ｜＝｜ＡＢ｜＋∑ｉ１＜ｉ２＜…＜ｉｔｊ１＜ｊ２＜…＜ｊｔ１≤ｔ≤ｍｎ－ｔ≥ｍＮＢｉ１ｉ２…ｉｔｊ１ｊ２…ｊｔＮＡＢ１…ｍ（－１）ｓｔ＋１ｊｔ＋１…（－１）ｓｎｊｎ其中ｉ１，ｉ２，…，ｉｔ是１，２，…，ｍ中ｔ个数码；ｊ１，ｊ２，…，ｊｔ，ｊｔ＋１，…，ｊｎ是１，２，…，ｎ的一个排列；ｓｒ＝π（ｊ１，ｊ２，…，ｊｔ，ｊｒ）（ｒ＝１，２，…，ｎ）是排列ｊ１，ｊ２，…，ｊｔ，ｊｒ的反序数．