饲料中糖/脂肪比例对瓦氏黄颡鱼生长、饲料利用、血糖水平和肝脏糖酵解酶活力的影响

以初始体重(4.20±0.02)g的瓦氏黄颡鱼(Pelteobagrus vachelli)幼鱼为实验对象,在流水系统中进行为期8周的摄食生长实验,探讨饲料中糖与脂肪比例(CHO∶L)对其生长、饲料利用、血糖水平和糖酵解酶活力的影响。饲料等氮(粗蛋白40%)等能(19 MJ/g),CHO∶L梯度为0.75—6.53。每种饲料随机投喂3桶鱼,每桶(50 L)放养40尾鱼。实验结果表明随着CHO∶L升高,瓦氏黄颡鱼特定生长率(SGR)先升高后降低,当CHO∶L为3.55时,SGR达最大,显著高于CHO∶L为0.75和6.53时的值(P<0.05)。饲料效率(FE)和蛋白效率(PER)在CHO∶L为1.30—3.55区间内都没有显著差异,但当CHO∶L为0.75或6.53时,FE和PER的值都显著降低(P<0.05)。脏体比(VSI)、肝体比(HSI)在各处理组间无显差异(P>0.05)。随CHO∶L的增加,全鱼粗蛋白含量先增加后降低,且在CHO:L为3.55时达到最大值,且显著高于CHO:L为0.75时的值(P<0.05)。而全鱼粗脂肪含量随CHO∶L增加而显著减少(P<0.05)。血糖和血浆总胆固醇水平在各处理组间无显著差异(P>0.05),而血浆甘油三酯水平随CHO∶L的增加而显著增加(P<0.05)。随CHO∶L的增加,瓦氏黄颡鱼肝脏葡萄糖激酶(GK)活力先增加后降低,且在CHO∶L为3.55时活力最大,显著高于CHO∶L为0.75时的值(P<0.05)。肝脏己糖激酶(HK)、丙酮酸激酶(PK)和磷酸果糖激酶(PFK-1)活力在各处理组间没有显著差异(P>0.05)。用二次多项回归模型拟合特定生长率和CHO∶L的关系,得到饲料最适CHO∶L为4.06。     

An enhanced protocol for expression and purification of heat-stable enterotoxin of enterotoxigenic Escherichia coli

We present an improved protocol for expression and purification of heat-stable enterotoxin (STa) of enterotoxigenic Escherichia coli (ETEC). In this protocol, controlled growth conditions at different pHs (7.4, 8.0, and 8.6) were adopted using a bioreactor. In addition, specific adsorbent resins, methacrylate, were used for STa purification. The bioreactor provided optimal ETEC growth at pH 7.4 with high STa production. Furthermore, methacrylate bounded specifically to STa and dramatically enhanced the purification process of STa. The STa-specific activity was high (8.9 × 10(6) units/mg protein), and the minimal effective dose of STa required for production of gut weight to remaining body weight ratio ≥ 0.083 was recorded as less than 0.2 ng in 2-3 days old suckling mice. The protocol presented, produces highly purified STa as documented by matrix-assisted laser desorption ionization-time of flight mass spectroscopy/. Also, as compared with the traditional methods, this procedure is trouble-free and practical for scale-up production and purification of STa peptides.     

Using matrix summation method for three dimensional dose calculation in brachytherapy

AimThe purpose of this study is to calculate radiation dose around a brachytherapy source in a water phantom for different seed locations or rotation the sources by the matrix summation method.BackgroundMonte Carlo based codes like MCNP are widely used for performing radiation transport calculations and dose evaluation in brachytherapy. But for complicated situations, like using more than one source, moving or rotating the source, the routine Monte Carlo method for dose calculation needs a long time running.Materials and methodsThe MCNPX code has been used to calculate radiation dose around a ¹⁹²Ir brachytherapy source and saved in a 3D matrix. Then, we used this matrix to evaluate the absorbed dose in any point due to some sources or a source which shifted or rotated in some places by the matrix summation method.ResultsThree dimensional (3D) dose results and isodose curves were presented for ¹⁹²Ir source in a water cube phantom shifted for 10 steps and rotated for 45 and 90° based on the matrix summation method. Also, we applied this method for some arrays of sources.ConclusionThe matrix summation method can be used for 3D dose calculations for any brachytherapy source which has moved or rotated. This simple method is very fast compared to routine Monte Carlo based methods. In addition, it can be applied for dose optimization study.     

Dynamics of the G protein-coupled vasopressin V2 receptor signaling network revealed by quantitative phosphoproteomics

G protein-coupled receptors (GPCRs) regulate diverse physiological processes, and many human diseases are due to defects in GPCR signaling. To identify the dynamic response of a signaling network downstream from a prototypical G(s)-coupled GPCR, the vasopressin V2 receptor, we have carried out multireplicate, quantitative phosphoproteomics with iTRAQ labeling at four time points following vasopressin exposure at a physiological concentration in cells isolated from rat kidney. A total of 12,167 phosphopeptides were identified from 2,783 proteins, with 273 changing significantly in abundance with vasopressin. Two-dimensional clustering of phosphopeptide time courses and Gene Ontology terms revealed that ligand binding to the V2 receptor affects more than simply the canonical cyclic adenosine monophosphate-protein kinase A and arrestin pathways under physiological conditions. The regulated proteins included key components of actin cytoskeleton remodeling, cell-cell adhesion, mitogen-activated protein kinase signaling, Wnt/β-catenin signaling, and apoptosis pathways. These data suggest that vasopressin can regulate an array of cellular functions well beyond its classical role in regulating water and solute transport. These results greatly expand the current view of GPCR signaling in a physiological context and shed new light on potential roles for this signaling network in disorders such as polycystic kidney disease. Finally, we provide an online resource of physiologically regulated phosphorylation sites with dynamic quantitative data (http://helixweb.nih.gov/ESBL/Database/TiPD/index.html).     

Engineering of chaperone systems and of the unfolded protein response

Production of recombinant proteins in mammalian cells is a successful technology that delivers protein pharmaceuticals for therapies and for diagnosis of human disorders. Cost effective production of protein biopharmaceuticals requires extensive optimization through cell and fermentation process engineering at the upstream and chemical engineering of purification processes at the downstream side of the production process. The majority of protein pharmaceuticals are secreted proteins. Accumulating evidence suggests that the folding and processing of these proteins in the endoplasmic reticulum (ER) is a general rate- and yield limiting step for their production. We will summarize our knowledge of protein folding in the ER and of signal transduction pathways activated by accumulation of unfolded proteins in the ER, collectively called the unfolded protein response (UPR). On the basis of this knowledge we will evaluate engineering approaches to increase cell specific productivities through engineering of the ER-resident protein folding machinery and of the UPR.     

Structure characterization and carboxymethylation of arabinoxylan isolated from Ispaghula (Plantago ovata) seed husk

As revealed by NMR spectroscopy (after ultrasonic degradation) and HPLC (after total hydrolysis) an arabinoxylan (AX) containing 74.8% Xylp and 23.2% Araf was isolated from Ispaghula (Plantago ovata) by soaking the seed husk with water, extraction with aqueous sodium hydroxide and coagulation with acetic acid. The AX with a molar mass of 364,470 g/mol shows high swelling ability in water. The carboxymethylation of AX was carried out heterogeneously with sodium monochloroacetate in the presence of aqueous sodium hydroxide. The reaction parameters were varied in terms of slurry medium, molar ratio, temperature, time, and sodium hydroxide concentration. For comparative studies, carboxymethylation of arabinan was carried out. In order to determine the total degree of substitution (DS) and mole fractions of the repeating units of carboxymethyl arabinoxylan (CMAX) and of carboxymethyl arabinan, HPLC and ¹H NMR spectroscopic investigations after total hydrolysis were carried out. DS values for CMAX as high as 1.81 were achieved. CMAX is water soluble starting at DS of 0.33.     

不同品种甘蔗茎尖细胞分裂节律研究

对 6个不同甘蔗品种 (粤糖 91 /976、粤糖 86/3 68、桂糖 1 1号、新台糖 1 6、CP80、农林 8号 )茎尖生长点细胞分裂进行切片观察研究 ,结果表明 :甘蔗一天内 (白天 )不同时间都存在细胞分裂 ,而且细胞分裂指数呈规律性变化 ,大部分呈先升后降再上升的变化趋势。除了桂糖 1 1号在 8∶0 0出现细胞分裂高峰之外 ,其它五个品种均在 1 0∶0 0出现细胞分裂高峰期 ,1 2∶0 0～ 1 4∶0 0是细胞分裂的低谷。种在不同生长时期细胞分裂指数变化规律不一样 ,细胞分裂高峰期出现的时期也不一样 ,早熟品种出现的时期早一些 ,晚熟品种则晚一些。甘蔗茎径和各生长时期细胞分裂指数是呈正相关的 ,茎径大 ,细胞分裂指数高 ,相反 ,茎径小 ,细胞分裂指数低。     

真核生物转录的延伸机理

由于对真核生物基因转录延伸期研究的忽视,现有的真核生物基因转录理论存在种种的偏颇.然而,新近的研究发现却使人们逐渐认识到,真核基因转录延伸阶段其实是真核细胞调节基因转录的一个高度有序、而又极为复杂的调控平台.本文尝试对近年来真核生物基因转录延伸领域的研究进展,包括转录延伸与mRNA加工的偶联、基因转录延伸调控的分子机理以及转录延伸调控对发肯和应激反应产生的影响,作概括性的综述.     

大孔树脂吸附分离烟草绿原酸的研究

通过比较8种大孔吸附树脂对烟草绿原酸的吸附分离性能,筛选出适合分离烟草绿原酸的树脂,并对其动态吸附特性进行研究.结果表明,XDA-1树脂对烟草绿原酸不仅吸附量大,而且解吸率高,适合烟草绿原酸的分离富集.该树脂吸附分离烟草绿原酸的工艺参数为:上柱液浓度3.5 mg/mL,pH 3.0,流速3倍柱床体积/h;以6倍柱床体积的40%乙醇进行洗脱,解吸附效果最佳,绿原酸总回收率为80.06%,初步吸附分离得到的产品中绿原酸含量为39.20 g/100 g.