小鼠过敏模型在食品过敏原分析中的研究与应用

建立牛乳蛋白、河虾蛋白的ICR小鼠过敏模型,检测食品过敏原体外激发小鼠致肥大细胞的组胺释放率,探讨食品过敏原鉴定并验证所建过敏原体外评价新方法。方法以牛奶蛋白、河虾蛋白,氢氧化铝佐剂免疫ICR小鼠,建立IgE高反应性食物过敏动物模型。间接ELISA测定激发后第7、14、21d血清中过敏原sIgE水平;并在激发第14d分离小鼠腹腔肥大细胞,过敏原体外激发,紫外分光光度法检测类胰蛋白酶定向释放水平。①模型小鼠血清sIgE水平明显高于对照组,且在过敏激发后第14dsIgE效价最高,河虾组和牛奶蛋白组的sIgE效价分别达到1/25 600和1/12 800。②牛奶组类胰蛋白酶释放率达38.5%;河虾组的类胰蛋白酶释放率达47.1%,所有模型组与对照组间差异均达显著水平(P<0.05)。建立了一种小鼠过敏模型,通过检测过敏原体外激发类胰蛋白酶释放率可以作为一种鉴定与评价食品过敏原的有效方法。     

过敏症豚鼠模型的建立及海虾主要过敏原组分的纯化与鉴定

旨在建立海虾过敏症豚鼠模型,分析主要过敏原蛋白质组分并将其应用ELISA来提高体外检测的准确性。以海虾蛋白浸液为致敏原,氢氧化铝为佐剂免疫豚鼠,建立豚鼠过敏模型,DEAE阴离子交换纯化主要的过敏原成分,间接ELISA法检测sIgE和IgG。实验结果:模型组血清sIgE和IgG效价分别为18∶0和12∶0480;DEAE阴离子交换层析成功纯化出了海虾中的主要过敏原成分,应用于ELISA检测sIgE效价为13∶20;主要过敏原蛋白检测sIgE效价是蛋白浸液用于检测的4倍。结果表明,海虾过敏症豚鼠模型建立成功,并且纯化的海虾过敏原组分应用于ELISA检测能提高sIgE的检测水平,对海虾过敏症的体外诊断和治疗有一定的意义。     

天津地区梭子蟹中过敏原组分分析

目的 分析天津地区梭子蟹蛋白提取液中主要过敏原组分特征.方法 常规方法制备蟹蛋白粗提液,经SDS-PAGE技术分析蛋白组分;分别以蟹过敏患者混合血清(sIgE)和单例血清(sIgE)为探针,应用Western-blot技术鉴别提取液中主要过敏原组分,同时观察不同患者过敏原组分的异质性.结果 蟹提取蛋白液SDS-PAGE结果显示,共有9条蛋白带,其中以70和74kD组分含量较高;Western-blot结果显示,除21、38kD外,其余多种蛋白组分均能与混合阳性血清反应,同时证实不同蟹过敏患者之间过敏原组分具有明显异质性.结论 天津地区蟹主要的过敏原是36、48、94kD三种蛋白,但不同的个体间主要过敏蛋白存在差异性.     

芝麻过敏原的分离、鉴定与纯化

通过SDS-PAGE电泳分离芝麻的蛋白质组份,采用免疫印迹(Westem-blotting方法其鉴定过敏原,通过离子交换层析对芝麻主要过敏原进行初步纯化.结果表明白芝麻粗提液SDS-PAGE显示有14条蛋白条带.黑芝麻粗提掖SDS-PAGE显示有16条蛋白条带.Western-Blotting显示对芝麻过敏患者的混合阳性血清均能与黑芝麻、白芝麻11 ku蛋白反应,离子交换层析可初步纯化出白芝麻11 ku的过敏原蛋白.     

中国对虾蛋白致敏小鼠模型构建及主要过敏原的双向电泳联合质谱鉴定

目的:构建中国对虾蛋白致敏小鼠模型并研究关键过敏原。方法:采用液氮研磨结合裂解液裂解对虾肌肉提取蛋白浸出液,用不同剂量的中国对虾蛋白和弗氏佐剂的混合液腹腔注射诱发ICR小鼠致敏,采用间接ELISA方法检测小鼠血清中过敏原特异性IgE和IgG1水平在激发期不同时间点的变化,并观察致敏小鼠行为评分,建立动物致敏模型;利用双向电泳分离虾蛋白,并与阳性血清免疫印迹,从而通过质谱分析其主要过敏原。结果:3组不同剂量的虾蛋白浸出液都能使小鼠出现不同程度的过敏症状,且剂量越高过敏反应越显著;其中,高剂量组(1.0 mg)小鼠血清特异性IgE和IgG1抗体含量显著升高(P0.05),出现瘙痒、嘴角红肿等现象,过敏症状最显著。通过双向电泳联合质谱鉴定发现,能与阳性血清反应的蛋白有两种,一种蛋白分子质量为40.2ku,等电点分别为6.0和6.2,经比对为精氨酸激酶;另一种蛋白分子质量为35.8 ku,其等电点为4.6,经比对为原肌球蛋白,通过Mascot数据库分析比较得到这两种蛋白质的氨基酸序列。结论:中国对虾蛋白可引起小鼠过敏反应,其主要过敏原为原肌球蛋白和精氨酸激酶,其中精氨酸激酶可能存在不同蛋白质修饰的两种形式。     

白果蛋白质提取及SDS-PAGE分析

以白果为原料,采用不同的原料处理及蛋白质提取方法,运用单因素和正交设计的方法,以料液比、提取时间、提取液浓度、提取液pH 值为考察因素,研究白果蛋白质提取的最佳工艺条件,并对提取的白果蛋白进行SDS-PAGE 凝胶电泳分析。结果表明：经过冷冻干燥处理的白果采用Tris-HCl 提取法获得的白果蛋白含量较高;Tris-HCl 法提取白果蛋白的最佳工艺为pH8.5、0.15mol/L Tris-HCl 溶液、料液比1:20(g/mL)、提取时间4h,白果蛋白质的提取率达到75.01%。白果蛋白SDS-PAGE 分析表明,白果蛋白中约含13 条亚基,主要为21kD 和32kD 的两种亚基,白果蛋白的亚基主要集中在31～100kD,占亚基总数的77%。     

腰果主要过敏原Ana o 2原核表达及免疫学鉴定

目的 构建Ana o 2重组表达质粒,原核表达重组蛋白并评价其免疫活性。方法 将Ana o 2基因构建到pET-28a(+)载体中,测序正确的重组质粒转化Rosetta(DE3)感受态细胞,诱导表达目的蛋白并进行质谱鉴定。利用腰果过敏阳性血清,通过酶联免疫吸附试验(ELISA)和蛋白质免疫印记(Western blot)法评价重组Ana o 2的免疫活性。结果 重组蛋白进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)表明分子量为54 kD,与理论值相符,经质谱鉴定为Ana o 2。ELISA结果显示,用重组Ana o 2检测腰果过敏阳性血清与阴性血清特异性IgE抗体(sIgE)水平差异有统计学意义(t=2.44,P<0.05)。Western blot结果表明,重组Ana o 2与腰果过敏患者血清反应性良好。结论 利用原核系统表达了Ana o 2,并且重组Ana o 2与腰果过敏患者血清具有良好的反应性。     

牛奶过敏原的分离、鉴定与纯化

对牛奶过敏原进行分离、鉴定与纯化。通过SDS-PAGE电泳分离牛奶的蛋白质组份,采用免疫印迹(Western-blotting)方法鉴定过敏原,通过离子交换层析对牛奶过敏原进行初步纯化。结果表明,鲜牛奶粗提液SDS-PAGE显示蛋白条带有12条,奶粉粗提液SDS-PAGE显示出的蛋白条带与鲜牛奶的蛋白条带基本一致。鲜牛奶Western-Blotting显示14ku的阳性条带。离子交换层析可初步纯化出14ku的过敏原蛋白。本研究对牛奶过敏原进行了分离和鉴定,并初步纯化出牛奶的主要过敏原。     

肥大细胞组胺体外释放模型在食品过敏原分析中的应用

目的：应用本室的肥大细胞组胺体外释放模型,分析食品过敏原中不同致敏组分的致敏差异性,验证建立的过敏原体外评价新方法。方法：牛奶、虾粗提物分别免疫C57／BL6小鼠,收集小鼠腹腔致敏肥大细胞（PMC）,分别用牛奶、虾不同过敏组分诱导PMC释放组胺,荧光法检测其释放水平。结果：牛奶中酪蛋白诱导牛奶致敏PMC释放组胺最多,脱脂乳和乳清次之,α-La则最少。酪蛋白是牛奶致敏蛋白中主要组分。虾致敏蛋白中,以80kD蛋白诱导组胺最多,36kD蛋白次之,21kD蛋白最弱,虾致敏蛋白混合诱导明显比单独诱导释放组胺多,说明80kD蛋白在虾致敏蛋白中起主要作用,并且各致敏蛋白间存在协同效应。结论：肥大细胞体外组胺释放量与过敏原种类及量存在相关性,本研究为进一步建立研究食品过敏和食品过敏原的新模型系统奠定基础。     

苦荞10 kD过敏原的重组表达及部分性质分析

以苦荞种子灌浆期cDNA文库中获得的苦荞10 kD过敏原基因序列TBAP10（tartary buckwheat 10 kD allergenprotein,TBAP10;GenBank登录号JK729379.1）为基础,构建重组表达载体pET47b-TBAP10,重组蛋白在大肠杆菌BL21 Star（DE3）中以包涵体形式表达。经包涵体复性和钴离子螯合层析纯化目的蛋白,并对其过敏活性、热稳定性及在模拟胃肠环境中的稳定性进行了分析。Western blotting显示该蛋白与苦荞16 kD过敏蛋白Fag t2存在免疫交叉反应。竞争性ELISA证明重组蛋白TBAP10具有与荞麦过敏患者血清IgE特异的结合活性;TBAP10具有强的热稳定性,能耐受15 min的沸水浴;模拟胃肠环境的消化结果显示TBAP10对胃蛋白酶具有强的耐受性,但对胰蛋白酶无耐受性。     

Textural Properties of Cold-set Gels Induced from Heat-denatured Whey Protein Isolates

A 9% whey protein (WP) isolate solution at pH 7.0 was heat-denatured at 80°C for 30 min. Size-exclusion HPLC showed that native WP formed soluble aggregates after heat-treatment. Additions of CaCl2 (10–40 mM), NaCl (50–400 mM) or glucono-delta-lactone (GDL, 0.4–2.0%, w/v) or hydrolysis by a protease from Bacillus licheniformis caused gelation of the denatured solution at 45°C. Textural parameters, hardness, adhesiveness, and cohesiveness of the gels so formed changed markedly with concentration of added salts or pH by added GDL. Maximum gel hardness occurred at 200 mM NaCl or pH 4.7. Increasing CaCl2 concentration continuously increased gel hardness. Generally, GDL-induced gels were harder than salt-induced gels, and much harder than the protease-induced gel.     

Occurrence of bisphenol-F-diglycidyl ether (BFDGE) in fish canned in oil

The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPL C with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether)analysed in the same products in the context of a previous study.     

Review of techniques to manufacture micro-hydrogel particles for the food industry and their applications

Microgels are ‘soft’ microscopic cross-linked polymeric particles that are being increasingly exploited in a variety of industries for rheology control, encapsulation and targeted delivery. They are valued because of the ability to tune their functionality to address specific applications in oil recovery, coatings, drug delivery, cosmetics, personal care and foods. Food microgels are typically biopolymer hydrogels in the form of microspheres, nanospheres (also called nanogels), spheroids and fibres. The utilisation of engineered microgels in foods has so far been limited, despite their great potential to address several needs in the food industry, including: satiety control, encapsulation of phytonutrients and prebiotics, texture control for healthier food formulations (e.g. reduced fat products), and targeting delivery to specific areas in the digestive tract. We review the scientific and patent literature on the utilisation and manufacturing methods for producing microgels with an emphasis on micro-hydrogels for food applications.     

12—A CRITIQUE OF SOME HYPOTHESES RELATING TO THE HETEROGENEITY OF THE HIGH-SULPHUR PROTEINS OF WOOL

Joubert and Burns prepared a large number of fractions from the high-sulphur proteins of wool and estimated their molecular weights and amino-acid compositions. Their data have been re-examined in order to look for statistically significant interrelations between amino acids and between the proportion of various amino acids and molecular weight. Statistical analysis of the data is also used to examine the credibility of some hypotheses concerning the mechanism of keratin biosynthesis and to provide further evidence for the existence of families of proteins within the high-sulphur fractions of wool.     

Contribution of European research to risk analysis

The European Commission's, Quality of Life Research Programme, Key Action 1—Health, Food & Nutrition is mission-oriented and aims, amongst other things, at providing a healthy, safe and high-quality food supply leading to reinforced consumer confidence in the safety of European food. Its objectives also include the enhancing of the competitiveness of the European food supply. Key Action 1 is currently supporting a number of different types of European collaborative projects in the area of risk analysis. The objectives of these projects range from the development and validation of prevention strategies including the reduction of consumers risks; development and validation of new modelling approaches; harmonization of risk assessment principles, methodologies, and terminology; standardization of methods and systems used for the safety evaluation of transgenic food; providing of tools for the evaluation of human viral contamination of shellfish and quality control; new methodologies for assessing the potential of unintended effects of genetically modified (genetically modified) foods; development of a risk assessment model for Cryptosporidium parvum related to the food and water industries; to the development of a communication platform for genetically modified organism, producers, retailers, regulatory authorities and consumer groups to improve safety assessment procedures, risk management strategies and risk communication; development and validation of new methods for safety testing of transgenic food; evaluation of the safety and efficacy of iron supplementation in pregnant women; evaluation of the potential cancer-preventing activity of pro- and pre-biotic ('synbiotic') combinations in human volunteers. An overview of these projects is presented here.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of     

The Latest Data about Development of Chinese Printing Industry in 2013

In the 2014 China（Shanghai）International Printing Week,Director Wang Yanbin released the latest data about development of Chinese printing industry in 2013.According to statistics,in 2013,the total output value of Chinese printing industry exceeded 1trillion Yuan for the first time,reaching 1.03985 trillion Yuan.There were 105,000 printing enterprises in China,employees were 3.415 million.The total asset was 1.06247 trillion Yuan;     

The State Council Issued "Directory of Government Approved Investment Projects(2013 Edition)"

正On December 2nd,2013,the State Council issued the notification of"Directory of Government Approved Investment Projects(2013 Edition)"(hereafter referred to as"notification").It is pointed out in the"notification"that in order to further deepen reforms in investment systems and administrative examination and approval systems,simplify administrative procedures and delegate powers to lower levels,earnestly     

China textile machinery: taking off in progress

正Among the 1600 exhibitors who take apart in the ITMA ASIA+CITME2014 2/3 are Chinese manufactures.If the numerous figures failed to attract your attention,the increase of quality should draw your focus.To adopt the demand of developing textile machine market,domestic textile machinery enterprises now follow the slogan of"technology drives development"to enhance product competitiveness.Our domestic sellers will showcase product ranging from spinning,weaving,dyeing and printing,     

Top Ten News of China's Paper Industry 2013

On December 24th, 2013, the meeting on the selection of top 10 news of China＇s paper industry 2013 sponsored by 〈China Paper Newsletters〉 was held in Beijing. The yearly selection of the top l0 news, which began in 2000, has become a brand activity widely recognized in the industry thanks to the support from the authorities at all levels and public participation.