中药复方提取物对槟榔所致大鼠急性炎症的影响

目的:减弱槟榔造成的炎症损伤，降低食用风险。方法:选择甘草、桔梗、枇杷叶3种中药材，以低、中、高剂量添加并与槟榔提取物共同灌胃大鼠28 d后，在大鼠足跖注射致炎剂，与空白对照组和阳性对照组大鼠比较体重、脏器系数、6 h内足跖肿胀率及炎症因子和炎症介质水平。结果:高剂量组相较于槟榔组可以显著(P<0.05)降低炎症介质前列腺素E₂(PGE₂)、丙二醛(MDA)和一氧化氮(NO)的产生，下调血清促炎因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6 (IL-6)水平以及促进白细胞介素-10 (IL-10)的释放，并减少肝脏中活性氧(ROS)的产生，且大鼠足跖肿胀率明显(P<0.05)低于空白对照组，与阳性对照组相近。结论:添加多种具有抗炎功效的天然植物提取物与槟榔共同使用可在一定程度上减轻因槟榔加剧的大鼠急性炎症。     

富含ACE抑制肽的酪蛋白水解物及其功能性酸奶对SHR血压的影响

目的:用酪蛋白双酶(胃蛋白酶和胰蛋白酶)水解物(A)及其超滤物(截留分子量6ku)(B)、水解物的脱苦产物(C)及其超滤物(D)分别灌胃12周龄雄性原发性高血压大鼠(SHR)和正常Wistar大鼠,研究其降血压作用.方法:A和C的灌胃剂量分别为0.083、0.25、0.75g/kg mb;B和D的灌胃剂量分别为0.01、0.03、0.09g/kg mb;检测灌胃后8h内大鼠尾动脉收缩压(SBP)的变化.结果:四种物质的高剂量对正常Wistar大鼠的血压无影响;四种物质的三种剂量分别灌胃SHR大鼠对其血压有明显的降低作用,其中A(0.25g/kg mb)、C(0.75g/kg mb)、B(0.03g/kg mb)、D(0.03g/kg mb)灌胃SHR大鼠在第4h达到最高降压幅度,分别为22.630、19.375、33.375、29.000mm Hg.用两种超滤物制备发酵乳灌胃SHR大鼠在第4h血压分别下降了36.875、31.875mm Hg;且间隔4h连续灌胃两种发酵乳可以在8h内维持较低的血压.结论:利用富含ACE抑制肽的酪蛋白水解物制备的功能性发酵乳在控制和治疗高血压方面有一定的效果,这为以后食源性降压食品的开发提供了一定的依据.     

超滤法分离免疫牛初乳中的IgG

本实验选用截留分子量为100kD的中孔纤维超滤膜从免疫牛初乳乳清中分离提取IgG,研究了操作压力和温度在超滤过程中对膜透过速率的影响,确立了最适操作压力为0.15MPa,最适操作温度为25℃;并采用分段超滤法对免疫牛初乳乳清中的IgG进行分离,从而使浓缩液中IgG纯度得到进一步提高。     

超滤去除砀山酥梨汁中色素和多酚的研究

本文研究超滤对砀山梨汁中色素和酚的去除作用.采用截留分子量分别为10、3和1k的聚醚砜（PES）超滤膜,对砀山酥梨汁进行过滤处理.10k膜和3k膜对色素有一定去除作用,1k对色素有显著去除作用,1k膜对色素的去除滤可达51%.三种膜对多酚的去除均有显著作用,去除率随截留分子量的减小而增加.超滤膜对色素和酚类的去除是筛分和吸附共同作用的结果.超滤对梨汁的某些理化指标有一定影响.再生可恢复超滤膜的性能.超滤对果汁中色素和酚类的去除能力使超滤在果汁加工中具有广泛的应用前景.     

不同分子量高阳离子度瓜尔胶控制废纸浆微胶黏物的特性

制备了高阳离子度瓜尔胶（HCG），并对其进行酸降解，制得不同分子量级别的HCG。研究了它们作为废纸浆微胶黏物固着剂控制微胶黏物含量的效果以及它们对纸页物理强度性能的影响，结果表明：HCG的分子量越高，越容易吸附于纤维；分子量越低，越容易与浆料中的溶解与胶体物质作用。尽管不同分子量级别的HCG的电荷密度远小于常用的聚胺类固着剂，它们去除浆内胶体物质和COD成分的效果却等同甚至好于后者，说明HCG的作用机理还包括氢键作用等其它方式。在废纸浆中加入一定量分子量适当的高阳离子度瓜尔胶，在有效控制微胶黏物含量的同时，能抑制纸张抗张强度和撕裂强度的下降，对耐折度有一定的增强作用。     

不同分子量残留对陈醋储存过程中沉淀物的影响

以6种滤膜进行超滤制备不同分子量残留的陈醋液,来探究陈醋中残留不同分子量物质对储存过程中沉淀物的影响。研究表明:陈醋液中残留不同分子量物质对陈醋的色度和感官评分影响不大,但残留物中蛋白质的平均疏水性与最终沉淀物的量呈显著正相关关系,相关系数为0.895。并且不同分子量物质在储存过程中发生沉淀的时间不同,其中大分子量物质(250kDa)在陈醋储存前120天发生沉淀。中分子量物质(150~250kDa)在储存120~150天范围内发生沉淀。小分子量物质(100kDa)在储存150天后产生沉淀,引起陈醋浊度的变化。     

鸡骨蛋白酶解液美拉德产物超滤组分的抗氧化性研究

通过超滤分级鸡骨蛋白酶解液美拉德产物(CBPH-MRPs),得到低组分(CM-I,分子量3ku),中组分[CM-Ⅱ,分子量(3 ku~10 ku)],高组分(CM-Ⅲ,分子量3 ku)。结果表明,褐色物质更多存在于CM-Ⅱ和CM-Ⅲ这些高组分中,而分子量对于中间产物影响不显著,荧光强度由强到弱依次为CM-ⅡCM-ⅢCM-I。经过测量抗氧化活性指标研究得出高分子量的MRPs抗氧化性较好,抗氧化性由强到弱依次为CM-ⅢCM-ⅡCM-I。     

芹菜籽提取物对高尿酸血症大鼠的影响

目的:研究芹菜籽提取物对于高尿酸血症大鼠血清尿酸水平的影响及对于急性痛风性关节炎的抗炎镇痛效果。方法:将50只Wistar雄性大鼠分为阴性对照组、模型对照组、芹菜籽提取物低、中、高剂量组。用酵母 腺嘌呤法致大鼠高尿酸血症,在造模的同时,给予上述药物灌胃,6w后检测各组大鼠血清尿酸(UA)、尿素氮(BUN)、肌酐(Cr)和肝脏黄嘌呤氧化酶活性等指标。肾脏进行病理检查。尿酸钠晶体致大鼠足跖肿胀,观察芹菜籽提取物抗炎作用。结果:高剂量组芹菜籽提取物可以降低高尿酸血症大鼠血清中尿酸的含量,差异有统计学意义(p<0.05)。中、高剂量组的黄嘌呤氧化酶活性低于模型对照组,但无统计学差异(p>0.05)。高剂量组足肿胀率明显低于模型对照组,差异具有统计学意义(p<0.05)。各剂量组肾脏病理改变轻于模型对照组。结论:提示芹菜籽提取物对于高尿酸血症及其引起的肾功能损害有一定的保护作用。     

大豆混合油中磷脂反相胶束分子量分布的测定

用截留分子量为1、2、10、20万的无机陶瓷膜超滤浓度为24.7%(W/W)的大豆混合油,测定了不同膜管截留分子量在不同超滤温度的磷脂截留率.超滤温度为50℃时,截留分子量1万的膜对磷脂的截留率为98.3%.分析了大豆混合油中磷脂反相胶束的分子量分布和温度的关系.温度越高,磷脂反相胶束的分子量越大,越有利于超滤脱胶的进行.     

超滤大豆蛋白肽对小鼠免疫功能的影响

目的:研究超滤大豆蛋白肽对小鼠免疫功能的影响。方法:以正常小鼠为实验对象,观察不同剂量(0.85g/kg.bw、1.7g/kg.bw、5.1g/kg.bw)的超滤大豆蛋白肽对小鼠脾脏指数、胸腺指数、小鼠迟发型变态反应(足跖增厚法)、半数溶血值和碳粒廓清指数的影响。结果:与对照组(30.25±2.12)相比,高剂量组和中剂量组的超滤大豆蛋白肽半数溶血值水平[(51.08±8.33),(38.29±5.51)]显著提高[(p<0.01),(p<0.05)];与对照组(4.90±0.63)相比,高剂量组和中剂量组超滤大豆蛋白肽吞噬指数[(5.78±0.69),(5.62±0.57)]显著提高(p<0.05);但各剂量组动物的脾脏指数、胸腺指数以及足趾厚度差无显著性差异(p>0.05)。结论:超滤大豆蛋白肽具有增强免疫功能中体液免疫功能和单核-巨噬细胞吞噬功能的作用。     

牛初乳与牛乳中乳清蛋白质组成及功能的对比分析

乳清富含多种功能特性和生物活性的蛋白质,本研究利用SDS-PAGE电泳将牛初乳与牛乳中乳清蛋白质的组成部分进行分离鉴定,发现牛初乳与牛乳中乳清蛋白质的组成存在较大的差异,且在牛初乳乳清中鉴定出290种蛋白,牛乳乳清中鉴定出325种蛋白。由GO功能注释分析发现,在生物过程中,牛初乳乳清蛋白在细胞定位建立和细胞定位中的作用略高于牛乳乳清蛋白。在分子功能上酶抑制活性作用是牛初乳乳清蛋白和牛乳中乳清蛋白的主要分子功能。在细胞组成上牛初乳乳清蛋白参与较多的是细胞外部分和细胞外空隙,与牛乳乳清蛋白相比参与的细胞组成大体相同。通过KEGG代谢通路分析可知,牛初乳和牛乳乳清蛋白均参与过补体及凝血级联反应通路。对牛初乳乳清蛋白组成进行研究,不仅能够增加牛初乳的利用率,并且为日后以乳清蛋白作为原料生产乳制品提供理论依据。     

Factors in ruminant colostrum that influence cell growth and murine IgE antibody responses.

Bovine colostrum was investigated as a source of biologically active molecules capable of stimulating the growth of mammalian cells in culture and modifying the immune response in a murine model. An extract prepared from bovine colostral whey by cation exchange and reversed-phase chromatography stimulated the growth of L6 rat myoblasts, Balb/c-3T3 mouse fibroblasts and BHK-21 baby hamster kidney cells with equal or greater potency than fetal bovine serum. Fractionation of the bovine colostral extract by gel-permeation chromatography in M-acetic acid identified a number of cell-growth factors for each cell type. Bovine colostral extract was compared with an ovine colostral whey preparation for its ability to modulate IgE antibody responses in mice. Doses of 8 and 4 mg/d of ovine colostral whey or bovine colostral extract specifically suppressed IgE antibody responses, whereas at lower doses suppression did not occur. We conclude that bovine colostrum contains cell-growth factors as well as immunomodulatory factors that are able to regulate the IgE response in a heterologous species.     

Assessment of adipogenic,antioxidant, and anti-inflammatory properties of whole and whey bovine colostrum

Bovine colostrum (BC) has been used for nutraceutical purposes for animals and humans. Bovine colostrum is a complex heterogeneous product and its antimicrobial activity, antioxidant potential, and growth factors can vary depending on age and species of the cow as well as their environment. Bovine colostrum preparation in skimmed or whey fractions can also alter properties of BC. Our goal was to compare cumulative anti-inflammatory, antioxidant, and adipogenic properties of natural (whole) versus whey BC. We compared properties of whole and whey BC in 3T3-L1 preadipocytes permanently transfected with reporters responding to changes in inflammatory (NfκbRE/green fluorescent protein), anti-inflammatory (Nrf2/YFP), and adipogenic (Fabp4/cyan fluorescent protein) status in cells. Interleukin-6 secretion in these cells was measured by ELISA. Whole and whey BC induce IL-6 secretion from 3T3-L1 fibroblasts; however, whey preparation stimulated less IL-6 secretion. Cumulative inflammatory nuclear factor (NF)κB activation in the presence of lipopolysaccharide was reduced by both whole (?27%) and whey BC (?22%) compared with lipopolysaccharide-treated cells (100%). Treatment with whole BC was more effective in the reduction of NFκB activation compared with whey BC and occurred in a dose-dependent manner. In consonance with decreased NFκB activation, the Nrf2 promoter activity was also reduced in response to whole (?27%) and whey (?13%) treatments compared with nontreated cells (100%). Whole and whey BC suppressed adipogenesis, measured as induction of Fabp4, by ?27 and ?13%, respectively, compared with nontreated 3T3-L1 fibroblasts (100%). Our results showed distinct differences in properties of whey and whole BC that could be used to attain reduced adipogenic or cumulative inflammatory responses.     

微滤-超滤联用技术优化牛初乳乳清中IgG富集工艺

为了高效富集IgG的同时减轻牛初乳的浪费问题,提高产品价值,本文采用微滤-超滤联用技术对牛初乳乳清中IgG进行富集。首先探究了微滤技术在牛初乳乳清除菌中的应用,并对其操作工艺进行优化,其次,利用超滤技术对微滤除菌后的牛初乳乳清进行富集,在单因素实验基础上,采用响应面对超滤工艺进行优化,并对富集后的牛初乳乳清进行品质分析。结果表明:牛初乳乳清微滤除菌的最佳工艺参数为:微滤压力为0.2 MPa、温度为30 ℃,超滤富集的最佳工艺参数为:超滤压力为0.15 MPa、温度为35 ℃、浓缩倍数为6倍、稀释次数为4次,按此条件进行牛初乳乳清的微滤-超滤操作,此时的IgG浓缩率为58.19%,膜通量为204.46 L/m2·h。富集后的牛初乳乳清品质分析表明:IgG含量为22760 μg/mL,IgG活性为718.31 IU/L,蛋白质含量为7.86%,脂肪含量为0.035%,菌落总数为2.4 lg CFU/mL。本研究为牛初乳乳清中IgG的进一步开发与综合利用提供了一定的参考依据。     

Milk growth factors as health products: Some technological aspects

Bovine milk and colostrum contain growth factors such as insulin-like growth factor IGF-I, IGF-II, transforming growth factor TGF-β1, TGF-β2, epidermal growth factor EGF, basic fibroblast growth factor bFGF and platelet-derived growth factor PDGF. A number of methodologies for the extraction of milk growth factors from milk, colostrum or whey have been developed. Cation-exchange chromatography has been widely used because of the basic nature of the growth factors. Also, microfiltration has been used for the concentration of some growth factors from colostrum, while ultrafiltration was successful only in separating IGF-I from IGF-II in whey. Growth factor extracts from milk, colostrum or whey have been used as therapeutic preparations for wound healing and in the treatment of inflammatory gut disorders. More recent applications are related to bone tissue regeneration and treatment of inflammatory skin diseases such as psoriasis.     

基于iTRAQ技术分析牛初乳与常乳乳清差异蛋白质组

为阐明牛初乳、牛常乳乳清蛋白的差异,利用同位素标记相对和绝对定量（isobaric tags for relative and absolute quantitation,iTRAQ）蛋白质组学技术对二者进行蛋白质组差异分析,在得到的599 种具有定量信息的乳清蛋白中,鉴定出60 种差异蛋白。将牛初乳与牛常乳乳清丰度差异蛋白进行生物信息学分析发现,差异蛋白主要参与的生物过程为转运、定位、单一生物作用等;主要参与的分子功能为顶端质膜、细胞外区域、细胞外区域部分等;主要参与的细胞组成为蛋白结合和阴离子结合。丰度差异蛋白中有9 种是与信号传导相关,有6 种糖基化乳清蛋白。此外,利用蛋白质网络互作分析发现,差异蛋白中存在具有高连接度的关键乳清蛋白因子。本研究采用iTRAO技术对牛初乳与牛常乳乳清差异蛋白进行鉴定及生物信息学分析,为今后改善牛初、常乳品质,开发婴幼儿乳粉以及功能性乳制品提供了一定参考。     

Enhancement of neutrophil function by ultrafiltered bovine whey

The objective of this work was to evaluate a proprietary ultrafiltered bovine whey product for its in vitro influence on the function of neutrophils from normal and dexamethasone-treated cattle, and for its in vivo influence on neutrophil function in periparturient dairy cows. The ultrafiltered bovine whey was produced by hyperimmunizing cows to various bacterial pathogens by intramammary injection, collecting and pooling the colostrum and milk for the first 3 wk after parturition, then separating and processing the whey through a filter with a nominal molecular mass cut off of 10,000 Da. In vitro treatment of neutrophils from normal calves with ultrafiltered bovine whey significantly increased neutrophil random migration, cytochrome C reduction, iodination activity, antibody-dependent cell-mediated cytotoxicity, and antibody-independent cell-mediated cytotoxicity. Cytochrome C reduction was the only neutrophil function parameter significantly enhanced by the in vitro treatment of neutrophils from dexamethasone-treated cattle with ultrafiltered bovine whey. In vivo treatment of periparturient cows with ultrafiltered bovine whey did not alter the total or differential leukocyte counts in the animals but did significantly increase the total erythrocyte counts. In vivo treatment with ultrafiltered bovine whey also significantly increased neutrophil iodination activity in the periparturient cows. Neutrophil iodination activity (a measure of the myeloperoxidase/hydrogen peroxide/halide antibacterial system) is a very potent bactericidal mechanism of neutrophils and has previously been shown to be suppressed in periparturient cows.     

Biological activity of bovine milk on proliferation of human intestinal cells

To evaluate the bioactivity of bovine milk from different stages of lactation on human intestinal tissue, a human fetal small intestinal cell line was used as a model system. Milk samples representing six stages of lactation: days 1, 2-3, 6-7 and weeks 12 and 24 after parturition, 1 week before drying off, and milk-like secretion from two stages of the dry period: 7 weeks and 3-4 weeks before expected calving, were collected from 64 Holstein Friesian cows. The whey fraction of the milk or milk-like secretion was added to the culture medium in concentrations ranging from 0.078% to 10%. The growth-promoting activity of whey was measured by determining the incorporation of [3H]thymidine into DNA for the last 24 h of the culture period. Whey fractions from all six stages of lactation stimulated growth of intestinal cells. The growth-promoting activity of colostrum or milk significantly decreased within the first week after calving. The growth-promoting activity in mature milk increased gradually during lactation to reach a level significantly higher than that obtained with colostrum. The growth-promoting activity of whey from milk-like secretion collected after drying off was lower than that of colostrum. Whey from different stages of lactation contained significantly different concentrations of TGF-beta1 (0.5-27 ng/ml) and TGF-beta2 (12-1219 ng/ml). However, neither the differences in TGF-beta1 and TGF-beta2, nor the differences in IGF-I and IGF-binding proteins could fully explain the differences in growth-promoting activity of colostrums or milk from different stages of lactation, suggesting that other factors were also involved. The present study showed that bovine milk contained a number of biologically active components that affected growth and development of human intestinal tissue. The results showed that the growth-promoting activity of colostrum and milk was dependent on the stage of lactation in accordance with previous results obtained with mammary epithelial cells. The changes in growth-promoting activity with stage of lactation were probably related to changes in concentrations of several growth factors.     

Bovine milk contains microRNA and messenger RNA that are stable under degradative conditions

We previously reported that microRNA (miRNA) is present in human breast milk. Recently, other groups have reported that bovine milk also contains miRNA; however, these reports are few. We therefore investigated bovine milk miRNA using microarray and quantitative PCR analyses to identify the differences between colostrum and mature milk. The RNA concentration in a colostrum whey fraction was higher than that in a mature milk whey fraction. In total, 102 miRNA were detected in bovine milk by microarray analysis (100 in colostrum and 53 in mature milk; 51 were common to both). Among these miRNA, we selected several immune- and development-related miRNA, including miR-15b, miR-27b, miR-34a, miR-106b, miR-130a, miR-155, and miR-223. These miRNA were detected in bovine milk by quantitative PCR, and each of these miRNA was significantly more highly expressed in colostrum than in mature milk. We also confirmed the presence of some mRNA in bovine milk. Nevertheless, synthesized miRNA spiked in the raw milk whey were degraded, and naturally existing miRNA and mRNA in raw milk were resistant to acidic conditions and RNase treatment. The RNA molecules in milk were stable. We also detected miRNA and mRNA in infant formulas purchased from Japanese markets. It is still unknown whether milk-derived RNA molecules play biological roles in infants; however, if milk-derived RNA do show functions in infants, our data will help guide future studies.     

Use of multi-angle laser light scattering and size-exclusion chromatography to characterize the molecular weight and types of aggregates present in commercial whey protein products

In this study, a range of commercial whey protein products were characterized by the use of size-exclusion chromatography coupled with a multi-angle laser light scattering (MALLS) detector. The MALLS system detected some very large-sized material that eluted close to void volume in all samples; this material was hardly detected by concentration detector. It was demonstrated by chitosan treatment that this peak was very small lipid globules or phospholipids, which gave the residual "cloudy" appearance in upper layers after ultracentrifugation of whey products. Composition, molecular weight, and the photo diode array (PDA) spectrum (200 to 400 nm) of the major protein peaks, including: beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), bovine serum albumin (BSA), immunoglobulin G (IgG) and some minor components were analyzed. The molecular weight of BLG, ALA, BSA, and IgG peaks in whey protein isolates (WPI) were 2.3 to 3.7 x 10(4), 1.4 to 1.6 x 10(4), 4.8 to 6.7 x 10(4), and 1.2 to 2.5 x 10(5) Da, respectively. Compared with WPI, WPC has similar major proteins, but more large-sized residual lipid material and different minor constituents such as lactose and nonprotein nitrogen, depending on various commercial samples and protein content. An improved TCA-precipitation method was applied to quantify glycomacropeptide (GMP) in whey proteins, which demonstrated that there was a very low concentration of GMP in WPI manufactured using an ion-exchange process. The molecular weight of GMP was found to be approximately 8600 Da. Size-exclusion chromatography MALLS was demonstrated to be a powerful technique for detailed analysis of the molecular weight of various proteins, aggregates and minor components, such as GMP, in whey protein products.