含酸奶饲料对衰老小鼠某些免疫功能指标的影响

用含Lactobacillus bulgaricus和Streptococcus Thermophilus活菌酸奶30％的饲料饲喂衰老小鼠3d，测定其胸腺重量、脾淋巴细胞增殖能力、IL-2（Interleukin，IL）产生量、脾T淋巴细胞中CD4^＋／CD8^＋细胞比例等指标，发现饲喂酸奶显著增加了胸腺重和IL-2的产生量,提高了脾T淋巴细胞的增殖能力，但未达到年轻小鼠的水平。此外还观察到酸奶处理提高了T淋巴细胞中CD4^＋／CD8^＋细胞的比例。这些结果表明，酸奶对于老年小鼠的免疫功能具有一定恢复作用。     

大豆异黄酮对青年雌性大鼠卵巢、子宫组织中Bcl-2 mRNA 和Bax mRNA 表达的影响

目的：通过动物实验研究,观察Bcl-2 mRNA 和Bax mRNA 在青年雌性大鼠卵巢和子宫组织中的表达情况,从细胞凋亡角度研究大豆异黄酮对青年雌性大鼠的抗衰老作用。方法：选用2 月龄青年雌性大鼠50 只,按体质量分成5 组,每组10 只,分别为对照组(基础饲料)、低剂量组(大豆异黄酮100mg/(kg bw·d))、中剂量组(大豆异黄酮200mg/(kg bw·d))、高剂量组(大豆异黄酮300mg/(kg bw·d))、雌激素组(己烯雌酚0.5mg/kg 饲料),实验周期7 周。采用原位杂交法检测各组大鼠卵巢和子宫组织中Bcl-2 mRNA 和Bax mRNA 的表达水平。结果：大豆异黄酮能够增强大鼠卵巢和子宫组织中Bcl-2 mRNA 的表达水平;而各剂量组大鼠卵巢和子宫组织中Bax mRNA水平呈下降趋势。结论：大豆异黄酮可以增强抗凋亡基因Bcl-2 mRNA,拮抗促凋亡基因Bax mRNA 的水平,推测这是大豆异黄酮对青年雌性大鼠发挥抗衰老作用的途径之一。     

纳豆激酶对大鼠酒精性肝损伤的改善效果及免疫调节作用

目的：探讨纳豆激酶对大鼠酒精性肝损伤的改善效果及免疫调节作用。方法：雄性Wistar大鼠随机分为5 组,正常对照组（生理盐水灌胃）、酒精模型组（酒精体积分数56%的白酒灌胃：第1周灌胃6 mL/（kg?d mb）,第2周灌胃8 mL/（kg?d mb）,第3周灌胃10 mL/（kg?d mb）,第4～10周灌胃11 mL/（kg?d mb））、纳豆激酶对照组（灌胃530.39 FU/（kg?d mb）纳豆激酶）、纳豆激酶干预组（灌胃530.39 FU/（kg?d mb）纳豆激酶＋酒精）、甘利欣干预组（灌胃200 mg/（kg?d mb）甘利欣＋酒精）;后两组酒精剂量同酒精模型组,实验持续10 周。苏木精-伊红染色观察肝脏组织形态结构,透射电子显微镜观察肝细胞超微结构;测定血清谷丙转氨酶（alanine aminotransferase,ALT）、谷草转氨酶（aspartate aminotransferase,AST）、γ-谷氨酰转肽酶（gamma-glutamyl transpeptidase,GGT）、胆碱酯酶（cholinesterase,CHE）活力;计算脾指数,流式细胞术检测大鼠外周血中CD4+、CD8+ T淋巴细胞亚群和自然杀伤（natural killer,NK）细胞比例。结果：酒精模型组大鼠肝组织形态结构和肝细胞超微结构均出现明显的病变损伤,纳豆激酶和甘利欣干预后得到显著改善。与酒精模型组相比,纳豆激酶干预组和甘利欣干预组大鼠血清ALT、AST、GGT活力显著下降（P＜0.05）,CHE活力有一定上升趋势。与酒精模型组相比,纳豆激酶干预组和甘利欣干预组大鼠脾指数、CD3+CD4+ T淋巴细胞比例、CD3+CD8+ T淋巴细胞比例均显著升高（P＜0.05）,TCRαβ+CD161a+ NK细胞比例有不同程度的上升趋势。结论：纳豆激酶对大鼠酒精性肝损伤有一定改善效果,其机制可能与调节CD4+、CD8+ T淋巴细胞、NK细胞等免疫细胞比例,提高机体免疫调节作用有关。     

复方大豆异黄酮钙软胶囊对大鼠骨密度的影响

目的：探讨复方大豆异黄酮钙软胶囊对去卵巢大鼠骨密度的影响。方法：将16周龄的雌性SD大鼠随机分为假手术对照组、去卵巢对照组和去卵巢低、中、高剂量共5组,其中3个剂量组每天分别给予复方大豆异黄酮钙软胶囊0.1、0.2、0.6g/kg·bw,连续灌胃12周后,检测各组大鼠的骨密度和骨钙含量。结果：复方大豆异黄酮钙软胶囊低、高剂量组大鼠股骨中点骨密度值显著高于去卵巢对照组（P〈0.05）;复方大豆异黄酮钙软胶囊中、高剂量组大鼠股骨远心端骨密度和股骨钙含量显著高于去卵巢对照组（P〈0.05）。结论：复方大豆异黄酮钙软胶囊具有增加去卵巢大鼠骨密度的功能。     

白藜芦醇改善γ射线诱导的T-淋巴细胞损伤大鼠的免疫功能

本文研究了白藜芦醇改善γ射线诱导的T-淋巴细胞损伤大鼠的免疫功能。选取SPF级50只SD健康大鼠，随机分为空白组、模型组、低剂量组（20 μmol/L）、中剂量组（40 μmol/L）和高剂量组（80 μmol/L），建立γ射线诱导的T淋巴细胞损伤模型大鼠。建模成功后，对大鼠进行白藜芦醇灌胃处理，按实验设计对大鼠细胞T细胞增殖率、T淋巴细胞核因子-kB（Nuclear factor kB，NF-kB）表达、T淋巴细胞（CD3、CD4、CD8）、干扰素-γ（Interferon-γ，IF-γ）、白介素-12（Interleukin-12，IL-12）、白介素-4（Interleukin-4，IL-4）、白介素-3（Interleukin-3，IL-3）水平进行检测。结果表明，中剂量组（40 μmol/L）白藜芦醇的T细胞增殖率、CD3、CD4、CD8、CD4/CD8分别为2.15%、54.15%、35.11%、21.45%、2.11。白藜芦醇对T细胞分泌的IF-γ、IL-12、IL-4、IL-3的水平分别为21546.15 pg/mL、212.15 pg/mL、645.56 pg/mL、221.15 pg/mL，与其他实验组均呈显著差异（p<0.05），说明白藜芦醇能够明显抑制调节T细胞，改善大鼠的免疫功能。     

大豆多糖对S180荷瘤小鼠抗肿瘤作用的免疫机制研究

从免疫学角度出发探讨大豆多糖的抗肿瘤作用。采用动物移植性肿瘤实验观察大豆多糖对小鼠体内肿瘤细胞生长和免疫器官胸腺、脾脏的影响;MTT法测定T淋巴细胞增殖功能;流式细胞仪检测大豆多糖对S180荷瘤小鼠T淋巴细胞亚群的影响;ELISA法检测荷瘤小鼠血清中细胞因子IL-2含量。结果表明大豆多糖对S180肉瘤生长有抑制作用,且以高剂量最佳;各剂量组对脾脏指数均有显著性提高,但中、高剂量给药组对胸腺有抑制作用;能提高小鼠脾淋巴细胞的增殖活性,与阴性对照组相比,中、高剂量组差异显著。大豆多糖明显增强ConA刺激的淋巴细胞增殖,与阴性对照组比较,中、高剂量组差异显著;能够不同程度地提高S180荷瘤小鼠脾细胞CD4+细胞数,增加CD4+/CD8+细胞比值;能提高S180荷瘤小鼠外周血血清中IL-2的含量。大豆多糖通过提高T淋巴细胞的增殖活性,并促进IL-2等细胞因子的分泌,由此发挥抗肿瘤作用。     

乳源酪蛋白糖巨肽对小鼠外周血T淋巴细胞亚群的影响

目的：酪蛋白糖巨肽（caseino-glycomacropeptide,CGMP）是κ-酪蛋白经凝乳酶和胃蛋白酶酶解后形成的含有64 个氨基酸的不同种类肽段,具有多种生物学活性和独特的营养特性,探讨CGMP对小鼠外周血T淋巴细胞亚群的影响。方法：以健康小鼠为实验动物,利用流式细胞仪分别检测长期和短期灌胃不同剂量的CGMP（30、60、120 μg/d）后,小鼠外周血淋巴细胞亚群的变化。结果：乳源CGMP干预后的结果表明,在短期灌胃实验中,灌胃不同剂量后CD3+淋巴细胞数比例均显著降低,12 h后恢复灌胃前水平;CD3+CD4+和CD3+CD8+淋巴细胞比例在灌胃30 μg和120 μg CGMP 4 h后均显著降低,而60 μg组则无明显变化;长期灌胃实验中,在第2天和第4天,外周血中CD3+淋巴细胞和CD3+CD4+比例与对照组相比显著升高（P＜0.05）;120 μg/d剂量组的小鼠外周血中CD3+CD8+淋巴细胞比例与对照组相比显著升高（P＜0.05）,而30 μg/d剂量组在2、4、6、8 d与对照组相比无显著变化（P＞0.05）;在第2天,30 μg/d和120 μg/d剂量组外周血中CD3+CD4+/ CD3+CD8+比例与对照组相比显著升高（P＜0.05）。结论：乳源CGMP能够引起正常小鼠的外周血淋巴细胞亚群发生变化,调节机体T淋巴细胞亚群的平衡,由此提示CGMP可作为功能性食品对外周血T淋巴细胞亚群的失衡有缓解作用。     

大豆异黄酮对糖尿病大鼠抗氧化能力的影响

目的：观察大豆异黄酮（soybean isoflavone，SIF）对糖尿病大鼠抗氧化能力的影响。方法：以四氧嘧啶（ALX）为诱导剂建立糖尿病动物模型，正常对照组（N）和糖尿病对照组（DC）以蒸馏水灌胃，大豆异黄酮组（SI）每日以100mg／kg·d的SIF溶液灌胃。实验6周后，分别测血清及肝脏中总抗氧化能力（T．AOC）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和黄嘌呤氧化酶（XOD）活性及丙二醛（MDA）含量。结果SI组实验后血糖较实验前和DC组明显降低（p〈0．05），DC组实验前血糖较实验后则明显升高（p〈0．05）。与DC组相比，SI组血清和肝脏GSH-Px和SOD抗氧化酶活性明显增强（p〈0．05）接近和超过正常水平，肝脏T．AOC能力明显高于DC和N组（p〈0．05），而血清及肝脏MDA含量均低于DC组（p〈0．05），血清XOD活性较DC组略有降低。结论：大豆异黄酮具有降血糖和增强机体抗氧化能力，其降糖机理与阻止胰腺B细胞和肝脏表面胰岛素受体进一步氧化损伤，修复其功能有关。     

嗜酸乳酸杆菌对自主活动受限型应激小鼠肠道黏膜免疫状态的影响

目的：探讨嗜酸乳酸杆菌（Lactobacillus acidophilus,LA）对自主活动受限型应激小鼠肠道黏膜免疫状态的影响。方法：以健康雌性BALB/c小鼠为受试动物,分为常规饲养组（NC组）、常规饲养条件灌胃LA组（NC＋LA组）、应激组（S组）、应激条件灌胃LA组（S＋LA组）,小鼠的LA灌胃剂量为108 CFU/d;应激组小鼠每天被限制自主活动3 h;15 d后取材,采用流式细胞术、酶联免疫吸附（enzyme-linked immunosorbent assay,ELISA）等方法,对肠系膜淋巴结（mesenteric lymph nodes,MLN）中CD4＋T细胞、CD8＋T细胞和CD11c＋树突细胞（CD11c＋ dendritic cell,CD11c＋ DC）比例,结肠组织白细胞介素-10（interleukin-10,IL-10）和白细胞介素-17（interleukin-17,IL-17）水平,以及小肠总分泌型免疫球蛋白A（secreted immunoglobulin A,sIgA）水平进行检测。结果：常规饲养条件下,LA能极显著提高小鼠MLN中CD4＋T细胞比例（P＜0.01）,显著提高MLN中CD11c＋ DC比例、小肠总sIgA水平和结肠组织IL-10水平（P＜0.05）,显著降低结肠组织IL-17水平（P＜0.05）,而对MLN中CD8＋T细胞比例无显著影响（P＞0.05）;应激条件能极显著降低小鼠MLN中CD4＋T细胞比例和CD11c＋ DC比例（P＜0.01）,显著降低CD8＋T细胞比例（P＜0.05）,极显著降低小鼠结肠组织IL-10水平和小肠总sIgA水平（P＜0.01）,极显著提高结肠组织IL-17水平（P＜0.01）;应激条件下,LA能显著提高小鼠MLN中CD4＋T细胞比例和CD11c＋ DC比例（P＜0.05）,显著提高小鼠结肠组织IL-10水平和小肠总sIgA水平（P＜0.05）,并显著降低结肠组织IL-17水平（P＜0.05）,而对小鼠MLN中CD8＋T细胞比例无显著影响（P＞0.05）。结论：给予自主活动受限型应激小鼠LA干预,可提高小鼠MLN中CD4＋T细胞和CD11c＋ DC比例,上调结肠IL-10和小肠总sIgA分泌水平,下调结肠IL-17分泌水平,调节应激小鼠的肠道黏膜免疫状态。     

白扁豆多糖对神经细胞缺氧性凋亡的保护机制

目的：模拟体内环境,研究白扁豆多糖（dolichos bean seed polysaccharide,DBSP）对胎鼠大脑 皮层神经细胞缺氧性凋亡的保护机制。方法：原代培养胎鼠大脑皮层神经细胞;建立缺氧/复氧（anoxia/ reoxygenation,A/R）损伤模型;实验分4 组：对照组、A/R组、白扁豆多糖预处理组（DBSP＋A/R组）、阻断剂组 （LY294002＋DBSP＋A/R组）;用比色法检测细胞活性与乳酸脱氢酶的释放水平;流式细胞仪分析测定Ca2＋ 相对浓度、活性氧、线粒体膜电位和细胞凋亡的变化;Western blot法检测总蛋白激酶B（total protein kinase B, T-Akt）和磷酸化蛋白激酶B（phosphorylated protein kinase B,p-Akt）蛋白表达。结果：从细胞活性来看,A/R组比 对照组明显降低（P＜0.01）,表明A/R对神经细胞产生了损伤。同时,与A/R组相比,DBSP＋A/R组细胞活性显著 上升、细胞凋亡明显减少（P＜0.01）,乳酸脱氢酶的释放水平下降,表明DBSP对受A/R损伤的神经细胞产生保护 作用,进一步结果显示,与A/R组相比,DBSP＋A/R组中Ca2＋相对浓度和活性氧生成显著降低、显著增加线粒体膜 电位和p-Akt蛋白表达（P＜0.01）。而与DBSP＋A/R组相比,LY294002＋DBSP＋A/R组中p-Akt蛋白表达显著减少 （P＜0.01）,同时细胞活性与线粒体膜电位下降,且LDH的释放水平、Ca2＋相对浓度、活性氧、细胞凋亡明显增加 （P＜0.05）,表明DBSP对神经细胞的保护作用受到PI3K-Akt信号通路抑制剂LY294002的抑制作用而被削弱。结 论：DBSP可通过PI3K-Akt信号转导通路抑制神经细胞的缺氧性凋亡。     

抗独特型抗体的研究进展

抗独特型抗体是针对抗体可变区的抗原决定簇（独特型）产生的特异性抗体，其中Ab2β是初始抗原在体内的“内影像”（intemal image），由于可模拟初始抗原而广泛应用于疫苗研究、肿瘤免疫、移植耐受、自身免疫病、食品安全等方面。对近年来抗独特型抗体的研究进展进行综述。     

辐照对虾过敏原抗原性的影响

目的探索辐照对海产品脱敏的效果,研究辐照对虾过敏原抗原性的影响。方法利用SDS-PAGE纯化的虾过敏原(原肌球蛋白)制备高效价、高特异性多克隆抗体,通过酶联免疫实验研究以3、5、7、10 kGy的辐照剂量处理对虾过敏原与患者特异性血清抗体IgE及多抗的免疫亲和力的影响。结果原肌球蛋白经辐照处理后,与患者特异性血清抗体IgE及鼠源多抗IgG的免疫亲和力下降。结论辐照可降低原肌球蛋白的致敏性。     

Immunological detection of moulds in food by using the enzyme-linked immunosorbent assay (ELISA); preparation of antigens

Antigens were isolated from moulds (Penicillium verrucosum var. cyclopium, Mucor racemosus and Fusarium oxysporum) by a simple water extraction followed by column chromatography using Sepharose CL4B. The antigens obtained were genus-specific, heat-stable and found not to be present in extracts of non-moulded food. Immunoglobulins were produced against these antigens and an enzyme-linked immunosorbent assay (ELISA) was developed. Using the ELISA, moulds were detected in both heated and unheated foods in a sensitive and specific way. The results indicate the possibility of testing whether prepared foods are produced from non-moulded raw materials or not. Heat treatment during processing does not influence the recovery of mould contamination.     

Binding of vitamin A with milk α- and β-caseins

The binding sites of retinol and retinoic acid with milk α- and β-caseins were determined, using constant protein concentration and various retinoid contents. FTIR, UV–visible and fluorescence spectroscopic methods as well as molecular modelling were used to analyse retinol and retinoic acid binding sites, the binding constant and the effect of retinoid complexation on the stability and conformation of caseins. Structural analysis showed that retinoids bind caseins via both hydrophilic and hydrophobic contacts with overall binding constants of K_{retinol-α-caseins} = 1.21 (±0.4) × 10⁵ M⁻¹ and K_{retinol-β-caseins} = 1.11 (±0.5) × 10⁵ M⁻¹ and K_{retinoic acid-α-caseins} = 6.2 (±0.6) × 10⁴ M⁻¹ and K_{retinoic acid-β-caseins} = 6.3 (±0.6) × 10⁴ M⁻¹. The number of bound retinol molecules per protein (n) was 1.5 (±0.1) for α-casein and 1.0 (±0.1) for β-casein, while 1 molecule of retinoic acid was bound in the α- and β-casein complexes. Molecular modelling showed different binding sites for retinol and retinoic acid on α- and β-caseins with more stable complexes formed with α-casein. Retinoid–casein complexation induced minor alterations of protein conformation. Caseins might act as carriers for transportation of retinoids to target molecules.     

免疫分析法检测无色孔雀石绿研究 Ⅰ人工抗原的合成

目的用免疫分析法检测水产品中无色孔雀石绿残留,研究无色孔雀石绿人工抗原的制备及其鉴定。方法用混酸法对无色孔雀石绿进行衍生化,连接上氨基基团,再经重氮化法与载体蛋白相结合,制备人工抗原。结果对无色孔雀石绿中间合成产物的液质联用分析结果表明,氨基产物中,无色孔雀石绿占51.83%,单氨基加成的无色孔雀石绿产物占40.48%,二氨基加成的无色孔雀石绿产物占7.69%。采用紫外法和Bradford法测定,人工抗原中无色孔雀石绿与载体蛋白的结合比为18∶1~23∶1。结论合成的人工抗原属于高质量的人工抗原。     

Interaction of milk α- and β-caseins with tea polyphenols

The interaction of α- and β-caseins with tea polyphenols (+)-catechin (C), (−)-epicatechin (EC), (−)-epigallocatechin (EGC) and (−)-epigallocatechin gallate (EGCG) was examined at a molecular level, using FTIR, UV–visible, CD and fluorescence spectroscopic methods as well as molecular modelling. The polyphenol binding mode, the binding constant and the effects of polyphenol complexation on casein stability and conformation were determined. Structural analysis showed that polyphenols bind casein via both hydrophilic and hydrophobic interactions with overall binding constants of K_C–α-cas = 1.8 (±0.8) × 10³ M⁻¹, K_EC–α-cas = 1.8 (±0.6) × 10³ M⁻¹, K_EGC–α-cas = 2.4 (±1.1) × 10³ M⁻¹ and K_{EGCG–α-cas} = 7.4 (±0.4) × 10³ M⁻¹, K_C–β-cas = 2.9 (±0.3) × 10³ M⁻¹, K_EC–β-cas = 2.5 (±0.6) × 10³ M⁻¹, K_EGC–β-cas = 3.5 (±0.7) × 10³ M⁻¹ and K_{EGCG–β-cas} = 1.59 (±0.2) × 10⁴ M⁻¹. The number of polyphenol bound per protein molecule (n) was 1.1 (C), 0.9 (EC), 1.1 (EGC), 1.5 (EGCG) for α-casien and 1.0 (C), 1.0 (EC), 1.1 (EGC) and 1.5 (EGCG) for β-casein. Structural modelling showed the participation of several amino acid residues in polyphenol–protein complexation with extended H-bonding network. Casein conformation was altered by polyphenol with a major reduction of α-helix and β-sheet and increase of random coil and turn structure suggesting further protein unfolding. These data can be used to explain the mechanism by which the antioxidant activity of tea compounds is affected by the addition of milk.     

Folic acid complexes with human and bovine serum albumins

The interaction of folic acid with human serum (HSA) and bovine serum albumins (BSA) at physiological conditions, using constant protein concentration and various folic acid contents was investigated. FTIR, UV–visible and fluorescence spectroscopic methods as well as molecular modelling were used to analyse folic acid binding sites, the binding constant and the effect on HSA and BSA stability and conformations. Structural analysis showed that folic acid binds HSA and BSA via both hydrophilic and hydrophobic contacts with overall binding constants of K_{folic acid–HSA} = 8.1 (±0.5) × 10⁴ M⁻¹ and K_{folic acid–BSA} = 1.0 (±0.3) × 10⁵ M⁻¹. The number of bound acid molecules per protein was 1.7 (±0.4) for HSA and 1.5 (±0.3) for BSA complexes. Molecular modelling showed participation of several amino acids in folic acid–protein complexes stabilised by hydrogen bonding network. Folic acid complexation altered protein secondary structure by major reduction of α-helix from 59% (free HSA) to 35% (acid-complex) and 62% (free BSA) to 25% (acid-complex) with an increase in random coil, turn and β-sheet structures indicating protein unfolding. The results suggest that serum albumins might act as carrier proteins for folic acid in delivering it to target molecules.     

Da vinci IntelliMap软件在特种纸机上的应用

介绍了Da vinci Intelli Map横向控制优化软件的技术特点,以及该软件在牡丹江恒丰纸业股份有限公司特种纸机上的应用情况。详细总结了Da vinci Intelli Map软件的各种功能、使用方法和注意事项。     

Interaction between whey proteins and lipids during light-induced oxidation

Dairy products are a suitable group for polyunsaturated fatty acid (PUFA) fortification, but are very susceptible to light-induced oxidation. In this study, the combined effect of light and PUFA fortification on the stability of milk proteins and on the formation of protein-lipid complexes was studied. Therefore, emulsions were prepared containing whey proteins and lipids with a different degree of unsaturation (olive, soybean, fish and algae oil). PUFA stimulated the protein oxidation process, even after complete degradation of the photosensitizer. First order reaction kinetics were established for riboflavin and tryptophan degradation, as well as the formation of N-formylkynurenine, a tryptophan degradation product. Oxidation was further characterised by degradation of the essential amino acids lysine, histidine, methionine and tyrosine. Protein-lipid adduct formation, measured fluorometrically and by the protein-bound carbonyl groups, could be linked with a decreased extractability of the unsaturated lipids from the emulsion, but also caused the formation of protein aggregates of high molecular mass.