恩施碎米荠硒多糖提取条件的研究

为了探索恩施碎米荠多糖的最佳提取条件,以干制碎米荠粉末为试验材料,以双蒸水为浸提剂,在浸提温度、时间、浸取剂的用量、提取次数和pH等单因素试验的基础上,进行了四元二次通用旋转组合试验.试验结果表明,1g碎米荠干粉,浸提温度90℃、浸提时间3 h-3.5 h、浸取剂的用量30mL、提取次数4～5次和浸提液的pH值为9是多糖适宜的提取条件.     

富硒壶瓶碎米荠对青鱼幼鱼生长、生理生化、硒代谢、抗氧化及先天免疫的影响

为探究富硒壶瓶碎米荠(Cardamine hupingshanensis)对青鱼(Mylopharyngodon piceus)幼鱼生长、血清生理生化、肝脏硒代谢、抗氧化能力和先天免疫指标的影响,选取360尾初始体重为(5.51±0.02) g的青鱼幼鱼随机分配至4个实验组中,每组3个重复。在基础饲料中添加富硒壶瓶碎米荠的量为0、0.5、1.0和2.0 g/kg(硒的实际含量分别为0.04、0.43、0.75和1.57 mg/kg),同时添加矿物质混合物(无硒添加)和维生素混合物,配置成4种等氮等能的青鱼幼鱼试验饲料,养殖周期为60d。结果显示:饲料中添加0.5和1.0 g/kg富硒壶瓶碎米荠时,鱼体增重率(WG),特定生长率(SGR)较对照组和过量组(2.0 g/kg)显著升高,饲料系数(FCR)显著降低(P<0.05)。当饲料中富硒壶瓶碎米荠添加量为0.5—1.0 g/kg时,血清中的甘油三酯(TG)、总胆固醇(TCH)和白蛋白(ALB)含量呈上升趋势,而葡萄糖(GLU)含量显著下降(P<0.05)。饲料中添加0.5—1.0 g/kg富硒壶瓶碎米荠时可显著提高肝脏核因子...     

一株耐硒壶瓶碎米荠内生菌分离、鉴定及其体外硒代谢研究

【背景】壶瓶碎米荠对硒具有超积累能力,并主要以硒代胱氨酸的形式存在,与已有的硒超积累植物显著不同,其硒超积累机制不明。【目的】从硒超积累植物壶瓶碎米荠(Cardamine hupingshanensis)体内分离耐硒内生菌,并对其进行鉴定和体外硒代谢特征研究,为壶瓶碎米荠超积累硒的机制研究提供参考。【方法】从壶瓶碎米荠新鲜叶片中分离纯化耐硒内生菌株,对其进行生理生化特征及16S rRNA基因序列分析鉴定,并对其进行亚硒酸钠培养代谢。【结果】获得一株耐硒内生菌CSN-1,被鉴定为甲基营养型芽孢杆菌(Bacillus methylotrophicus),培养液中硒含量低(Se 1.5 mg/L)时其吸光度值较对照组高,硒含量高(Se 10 mg/L)时其吸光度值较对照组低;代谢后的上清液中硒主要以Se~(4+)存在,而菌体中硒主要是硒代胱氨酸(SeCys_2)。【结论】硒超积累植物壶瓶碎米荠叶片体内存在甲基营养型芽孢杆菌(Bacillus methylotrophicus)CSN-1,具有将亚硒酸钠转化为硒代胱氨酸的能力,低浓度的硒对该内生菌的生长具有一定的促进作用,而高浓度的硒则会抑制该内生菌的生长。     

富硒产品中硒的形态分析及化学评分模式的建立

采用微波水解、HPLC-HG-AFS法测定了硒蛋白粉、硒蛋白片、肽粉、富硒原料等19种硒产品中的总硒、硒代氨基酸和亚硒酸根离子[Se(IV)],分析了硒代氨基酸、Se(IV)和其他形态硒占总硒的百分比及不同形态硒代氨基酸的组成比例。以此为依据,将硒产品分为硒蛋白型、单一硒代氨基酸型、其它形态硒型及有机无机硒混合型。根据DBS42/002-2014规定建立了富有机硒产品评分模式,其中18种为富有机硒产品;根据适硒地区母乳中硒代氨基酸的组成比例提出了硒代氨基酸的化学评分模式,评分结果显示13种以蛋白态硒为主的硒产品中硒代氨基酸的组成比例均与母乳相差甚远,不利于人体平衡吸收利用,其中10种SeMet含量远远超过人体所需,SeCys_2为限制硒代氨基酸。该评分模式的建立对硒产品的开发具有指导意义。     

基于生态位理论的富硒土壤资源开发利用适宜性评价及分区方法

实施乡村振兴战略,产业兴旺是关键。立足区域富硒土壤资源禀赋优势,发展富硒农业特色经济,是振兴乡村产业的一条有效途径。以多维超体积生态位理论为指导,以区域富硒土壤资源禀赋和富硒农业发展要求为基础,从土壤自身条件、土地利用和生态保护3个方面明确了富硒土壤资源开发利用的主导因素、区位因素、限制因素;选择土壤硒含量、土壤有机质含量、地形坡度、土地利用类型、土地综合污染指数、生态保护建设等6个关键因子,构建了富硒土壤资源开发利用生态位适宜性评价指标体系;利用生态位适宜性评价相关模型,探讨了生态位适宜性单一指数和综合指数的计算方法;结合土壤利用改良分区、土壤利用功能分区、富硒农业生产分区等,分析了区域富硒土壤资源开发利用适宜性分区问题。研究可为富硒土壤资源开发利用适宜性评价提供科学可行的思路与方法,对于促进区域富硒土壤资源科学合理利用、实现富硒农业产业科学发展具有重要的指导意义和现实意义。     

砂藓总蛋白质提取方法的选取及双向电泳体系的建立

为进行砂藓(Racomitrium canescens)蛋白质组学的双向电泳工作,首要前提是获得质量较好的砂藓总蛋白质,并建立一套完整高效的适用于砂藓的双向电泳体系。本研究以砂藓为研究对象,比较了Tris-酚抽提法、TCA-丙酮法和试剂盒法3种蛋白质提取方法获得砂藓总蛋白质的浓度和完整性,并对砂藓中蛋白质双向电泳的运行条件进行了优化和比较。结果表明:在3种砂藓总蛋白质提取方法中,Tris-酚抽提法获得的砂藓总蛋白质浓度最高,SDS-PAGE电泳结果中条带清晰且较完整,双向电泳后获得的蛋白质点最多,是3种方法中提取砂藓蛋白质浓度最高完整性最好的方法;基于Tris-酚抽提法的基础上进一步对砂藓总蛋白质双向电泳体系进行优化,结果显示:利用pH值范围为pH 4～7的IPG胶条,蛋白质上样量为1 000μg的条件下,获得的砂藓蛋白质双向电泳图谱蛋白质点最多,图谱中可分辨的蛋白质点共(872±15)个,且低丰度蛋白质点清晰。本研究筛选出的方法获得了蛋白质点分布均匀、背景清晰、分辨率高、质量较高的双向电泳图像,建立了砂藓蛋白质组学研究体系的较好方法。本研究结果为砂藓蛋白质组学研究提供了良好基础。     

动胶菌发酵产物PHAs提取方法的研究

以动胶菌为生产菌株,对后期提取方法进行筛选,从而确定了提取聚羟基烷酸酯(PHAs)的最佳方案即季铵盐浓度为1%,作用时间为15 min,细胞浓度为350 g/L,次氯酸钠浓度为3%,作用时间为5 min.以该方案为基础的放大实验,得到产物纯度98.3%;经力学性质测试拉伸强度为26.265 MPa;杨氏模量1287.535 MPa;经热分析测试熔点为173℃,熔解焓43.18Jg- 1.     

茶叶中咖啡因提取实验方法的比较及改进

“茶叶中咖啡因的提取”实验是有机化学中的一个重要实验,特别是对工科的化学工程、生物工程、生物制药专业来说,更是必做不可。但由于该实验耗时长,实验过程有一定难度,不易在教学课时内完成,过去也经常听到有同行说实验做不成功的。为此,一方面为了提高实验成功率,另一方面为了简化实验方法,方便学生操作,我们对现有的多种提取方法进行比较,以期为今后的实验设计能够提供指导作用。     

红树植物秋茄叶片双向电泳技术体系的建立及优化

对适用于秋茄(Kandelia candel)叶片蛋白质组学研究的双向电泳技术体系进行了优化.结果表明,采用酚抽提法提取的蛋白质浓度较低,约1.7 μg μL-1,SDS-PAGE电泳后几乎无条带;而采用改良TCA -丙酮沉淀法可显著提高提取液中蛋白质的含量,可达12.4 μgμL-1.秋茄叶片蛋白质主要分布在pH4～...     

盐肤木基因组DNA提取方法改进及AFLP体系的建立

经过反复试验,摸索出一种提取高质量植物基因组DNA的方法:改良的4×CTAB法.以盐肤木叶片为实验材料,提取到高质量的基因组DNA,建立了酶切、连接、预扩增、选择性扩增的AFLP反应体系.通过两种引物组合"E+3/M+3"和"E+2/M+3"策略筛选出8对条带分辨率高、多态性好的引物组合,优化了盐肤木的AFLP银染反应...     

黄粉虫有效物质的综合提取及提取方法的比较

比较了用石油醚和用环己烷抽提黄粉虫TenebriomolitorL .虫粉中脂肪的提取率 ,以及用碱法和用酶法提取去脂虫渣中蛋白质的提取率。并从 2种去脂、去蛋白质虫渣中和虫蜕中提取壳聚糖。用环己烷和酶法提取为最佳组合 ,可综合提取出占幼虫干重 5 0 .5 %的有效物质。同时 ,讨论了碱法和酶法提取率出现差异的原因。     

Evaluation of methods for total selenium determination in yeast

The selenium determination in biological materials by the classical fluorometric method (FM) is time-consuming and also hazardous, as it requires the destruction of the organic matrix samples with hot HNO₃/HClO₄ mixtures prior to analysis. Accordingly, commercial analytical laboratories are increasingly using faster instrumental methods; for sample digestion, avoid using HClO₄. Because of these procedural changes, the results obtained by commercial laboratories may be unreliable, especially for samples containing Se in organic forms. One such “difficult” substrate is Se yeast, which contains most of its Se as selenomethionine. To establish which methods for Se analysis and sample digestion are applicable, samples of Se yeast and of selenomethionine standards were sent to laboratories employing either flame atomic absorption spectrometry (FAAS), inductively coupled plasma-mass spectrometry (ICP-MS), or hydride generation atomic absorption spectrometry (HGAAS). The result were compared with those obtained by FM and non-destructive instrumental neutron activation analysis (INAA). ICP-MS, after microwave digestion of sample with HNO₃/H₂O₂, produced results within 5% of the expected values, as did those obtained by FM and INAA. With FAAS, acceptable results were obtained after digestion with HNO₃/HCl. With HGAAS, sample digestion with HNO₃/H₂O₂ produced values that were systematically elevated by about 10% and exhibited standard deviations of ≥10%. Thus, current methods of sample digestion are applicable for Se yeast analysis by ICP-MS and FAAS, but not by HGAAS.     

Serum selenium and single-nucleotide polymorphisms in genes for selenoproteins: relationship to markers of oxidative stress in men from Auckland, New Zealand

There is controversy as to the recommended daily intake of selenium (Se), and whether current New Zealand diets are adequate in this nutrient. Various functional single-nucleotide polymorphisms (SNPs) polymorphisms may affect the efficacy of Se utilisation. These include the glutathione peroxidases GPx1 rs1050450, GPx4 rs713041, as well as selenoproteins SEPP1 rs3877899, SEL15 rs5845, SELS rs28665122 and SELS rs4965373. This cross-sectional study measured serum Se levels of 503 healthy Caucasian men in Auckland, New Zealand, between ages 20–81. The Se distribution was compared with activities of the antioxidant enzymes glutathione peroxidase and thioredoxin reductase, and DNA damage as measured by the single cell gel electrophoresis assay, both without and with a peroxide-induced oxidative challenge. Serum Se was measured using inductively coupled plasma-dynamic reaction cell-mass spectrometry, while selenoprotein SNPs were estimated using TaqMan^® SNP genotyping assays. While antioxidant enzyme activities and DNA damage recorded after a peroxide challenge increased with increasing serum selenium, the inherent DNA damage levels in leukocytes showed no statistically significant relationship with serum selenium. However, these relationships and dietary Se requirements at the individual level were modified by several different SNPs in genes for selenoproteins. The GPx1 rs1050450 C allele was significantly associated with GPx activity. Significant correlations between serum Se level and GPX activity were seen with all genotypes except for homozygous minor allele carriers, while the GPx1 rs1050450 CT genotype showed the highest correlation. Several genotypes showed significant correlations between serum Se and TR activity with SEPP1 rs3877899 GG genotype showing the highest correlation. A significant decreasing trend in DNA damage with increasing serum Se was seen among GPx1 rs1050450 CC and GPx4 rs713041 TT genotype carriers up to a serum Se level of 116 and 149 ng/ml, respectively. In the absence of this genetic information, we would recommend a serum Se concentration in the region of 100–150 ng/ml as providing a useful compromise.     

双磁珠法质粒DNA提取技术及应用

质粒是基因合成与测序领域中使用最为频繁的基因运载工具,然而传统的质粒DNA提取方法面临提取通量低、生产成本高等问题,无法满足日益增长的需求。本研究基于质粒提取原理,开发了双磁珠法(double-magnetic-bead method, DMBM)质粒提取技术,探究了磁珠投入量、质粒DNA片段大小、菌液投入量等因素对质粒提取的影响,并且对比了本技术与商业化质粒DNA提取试剂盒提取DNA质量、提取通量及提取成本。结果表明,双磁珠法质粒DNA提取技术可满足不同细胞密度、不同片段长度的质粒DNA提取。此外,该技术搭载96通道全自动核酸提取仪,提取的质粒DNA纯度更高、提取时间缩短80%、提取成本缩减57.1%,从而实现了质粒DNA提取的高通量、低成本,有效助力基因合成与测序。     

超临界CO_2萃取黄绿卷毛菇脂肪酸成分的GC-MS分析

超临界CO2萃取黄绿卷毛菇所得产物进行了GC-MS分析,分离和鉴定了25种脂肪酸的成分和含量,其中亚油酸相对含量为10.6%,单不饱和脂肪酸相对含量占31.5%,包括反式油酸、顺式油酸、反式-10-羟基-6-甲氧基-十八烯(10)酸、顺式-10-羟基-6-甲氧基-十八烯(10)酸、反式-11-羟基-5-甲氧基-十八烯(11)酸、顺式-11-羟基-5-甲氧基-十八烯(11)酸、7-甲氧基-11-羟基-二十烯(11)酸和6-甲氧基-12-羟基-二十烯(12)酸等,饱和脂肪酸相对含量占56.9%。     

Seasonal fluctuations of selenium and sulfur accumulation in selenium hyperaccumulators and related nonaccumulators

Some plants hyperaccumulate selenium (Se) up to 1% of dry weight. This study was performed to obtain insight into whole-plant Se fluxes in hyperaccumulators. Selenium hyperaccumulators Astragalus bisulcatus and Stanleya pinnata were monitored over two growing seasons for seasonal fluctuations in concentrations of Se and the chemically similar element sulfur (S). The related nonhyperaccumulators Astragalus sericoleucus, Oxytropis sericea and Thlaspi montanum were included for comparison. In both hyperaccumulators leaf Se decreased from April to October, coinciding with Se hyperaccumulation in flowers and seeds. Root Se levels were lowest in summer. Selenium concentration decreased with leaf age in both hyperaccumulators. Leaf S levels peaked in summer in all plant species, as did Se levels in nonhyperaccumulators. Selenium and S levels tended to be negatively correlated in hyperaccumulators, and positively correlated in nonhyperaccumulators. These results suggest a specific flow of Se in hyperaccumulator plants over the growing season, from root to young leaves in spring, followed by remobilization from aging leaves to reproductive tissues in summer, and back to roots in the autumn.     

Detection and quantification of methicillin‐resistant Staphylococcus aureus (MRSA) clones in retail meat products

Aims: The objective of the study was to determine the prevalence of methicillin‐resistant Staphylococcus aureus (MRSA) contamination of retail meat and to determine the level of contamination. Methods and Results: Pork (pork chops and ground pork), ground beef and chicken (legs, wings and thighs) were purchased at retail outlets in four Canadian provinces and tested for the presence of methicillin‐resistant Staph. aureus using qualitative and quantitative methods. MRSA was isolated from 9·6% of pork, 5·6% of beef and 1·2% of chicken samples (P = 0·0002). Low levels of MRSA were typically present, with 37% below the detection threshold for quantification and <100 CFU g^?1 present in most quantifiable samples. All isolates were classified as Canadian epidemic MRSA‐2 (CMRSA‐2) by pulsed field gel electrophoresis (PFGE), with two different PFGE subtypes, and were spa type 24/t242. Conclusions: MRSA contamination of retail meat is not uncommon. While CMRSA‐2, a human epidemic clone, has been found in pigs in Canada, the lack of isolation of livestock‐associated ST398 was surprising. Significance and Impact of the Study: The relevance of MRSA contamination of meat is unclear but investigation is required because of the potential for exposure from food handling. Sources of contamination require investigation because these results suggest that human or animal sources could be involved.     

Immunomodulatory effect of selenosemicarbazides and selenium inorganic compounds, distribution in organs after selenium supplementation

Antioxidant properties of selenium producing a protective barrier against free radicals play an important role in numerous metabolic and immunologic processes associated with oxidation- reduction reactions which take place during intracellular digestion of phagocyted bacteria. The aim of our study was to examine the properties of an organic compound of selenium, 4-(o-tolilo)- selenosemicarbazide of p-chlorobenzoic acid in terms of its retention in organs, effect on erythropoesis and phagocytic abilities of neutrophiles as well as antioxidant properties in neutrophiles tested with NBT test. This compound as well as inorganic sodium selenate was given to Swiss mice at the dose of 10^–3 g Se/kg for the period of 10 days. The concentrations of selenium in livers of mice treated with sodium selenate and selenosemicarbazide were found to be higher than in controls (18,7 g lg^–1 and 23.2 g lg^–1 vs. 12 g lg^–1, respectively). Analysis of blood cells count has shown a significant decrease in neutrophile levels in both groups treated with selenium. The influence of selenium compounds on phagocytosis and especially NBT test has been determined (3.8% of positive cells in the controls vs. 2.2% and 0.9% in the groups treated with sodium selenate and selenosemicarbazide, respectively). Our preliminary investigations suggest that selenosemicarbazides are biologically active compounds and can modify neutrophile functions.     

不同工艺提取老鹰茶总黄酮含量的比较

比较不同工艺提取富集的老鹰茶中总黄酮含量,分别采用传统乙醇提取-正丁醇萃取法和水提取-大孔树脂法提取老鹰茶总黄酮,并用紫外分光光度法,以芦丁为标准品,270nm下测定总黄酮含量。水提取-大孔树脂法富集所得固体中黄酮含量较高,可达55.86%,回收率为105.6%,RSD=1.64%。水提取-大孔树脂富集的工艺操作简单,所得固体黄酮含量高,更适于老鹰茶总黄酮的提取,对进一步的工业化生产开发有较好的参考价值。     

牛脾转移因子的提取工艺研究

目的：比较从牛脾中提取活性ＴＦ的不同方法。方法：分别利用Ｌａｗｒｅｎｃｅ－－透析法、Ｌａｗｒｅｎｃｅ－超滤法、超离心－超滤法提取,再以蛋白反应、红外及紫外扫描予以鉴定比较。结果：Ｌａｗｒｅｎｃｅ－超滤法和超离心－超滤法效果较好。结论：工业化生产中应采用Ｌａｗｒｅｎｃｅ－超滤法。