Serum biomarker profiling in cancer studies: a question of standardisation?

Companion animals are exposed to similar environmental conditions and carcinogens as humans. In some animal cancers, there also appears to be the same genetic changes associated as in humans. However, little work has been carried out in cancer biomarker identification in animals. The recent dramatic advances in molecular medicine, genomics, proteomics and translational research will allow biomarker identification, which may provide the best strategies for veterinarians and clinicians to combat disease by early diagnosis and administration of effective treatments. Proteomics may have important applications in cancer diagnosis, prognosis and predictive clinical outcome that could directly change clinical practice by affecting critical elemen‐ts of care and management. This review summarizes the advances in proteomics that has propelled us to this exciting age of clinical proteomics, and highlights the future work that is required for this to become a reality. In this review, we will discuss the available proteomic technologies and their limitations, and highlight the key areas of research and how they have been used to discover cancer biomarkers. The principles described here are equally applicable to human and animal disease, but implementation of ‘omic’ technologies requires stringent guidelines for collection of clinical material, the application of analytical techniques and interpretation of the data.     

柔嫩艾美耳球虫棒状体颈部蛋白2基因真核表达载体的构建及在293T细胞中表达

为构建柔嫩艾美耳球虫棒状体颈部蛋白2(Eimeria tenella rhoptry neck protein 2,EtRON2)基因的重组质粒pCAGGS-EtRON2,并转染293T细胞进行表达,以柔嫩艾美耳球虫孢子化卵囊cDNA为模板,经PCR扩增其核心编码区的一部分,将其克隆至pGEM-Teasy载体,构建pGEM-Teasy-EtRON2质粒,双酶切出目的片段后与相应酶切的真核表达载体pCAGGS连接,构建真核表达质粒pCAGGS-EtRON2。该重组质粒经酶切和测序鉴定后转染293T细胞进行表达,分别用免疫印迹和间接免疫荧光鉴定EtRON2基因的表达情况。所构建的真核表达质粒pCAGGS-EtRON2经过双酶切鉴定,可见一条大小约为1172 bp的目的条带,测序结果与GenBank所登录序列完全一致;免疫印迹实验可见大小约为43 kDa的目的蛋白条带,间接免疫荧光实验可以检测到特异性红色荧光。研究结果表明已成功构建了EtRON2的真核表达质粒pCAGGS-EtRON2,并在真核细胞中获得表达,为深入研究EtRON2的生物学功能奠定了基础。     

饲粮蛋白质水平对冬毛期水貂胃肠道消化酶活性以及空肠形态结构的影响

本文旨在研究饲粮蛋白质水平对冬毛期水貂胃肠道消化酶活性以及空肠形态结构的影响。选择 ９０日龄的健康公貂 ６０只,随机分成 ６组（每组 １０个重复,每个重复 １只）,分别饲喂蛋白质水平为 ２８％（Ⅰ组）、３０％（Ⅱ组）、３２％（Ⅲ组）、３４％（Ⅳ组）、３６％（Ⅴ组）和 ３８％（Ⅵ组）的试验饲粮。预试期 ７ｄ,正试期 １１５ｄ。饲养试验结束后进行屠宰,制备胃肠道内容物上清液,用于测定消化酶活性;制备空肠组织切片,用于观察空肠的形态结构。结果表明：Ⅴ组水貂的胃蛋白酶活性极显著高于Ⅰ、Ⅱ和Ⅲ组（Ｐ＜０．０１）,与Ⅳ和Ⅵ组差异不显著（Ｐ＞０．０５）。Ⅳ组水貂的十二指肠和空肠胰淀粉酶活性极显著高于Ⅰ、Ⅱ、Ⅲ、Ⅴ和Ⅵ组（Ｐ＜０．０１）。Ⅳ和Ⅴ组水貂的回肠胰淀粉酶活性显著或极显著高于Ⅰ和Ⅱ组（Ｐ＜０．０５或 Ｐ＜０．０１）。Ⅳ组水貂的十二指肠胰蛋白酶活性极显著高于Ⅰ、Ⅱ、Ⅴ和Ⅵ组（Ｐ＜０．０１）,空肠胰蛋白酶活性极显著高于Ⅰ、Ⅱ、Ⅲ、Ⅴ和Ⅵ组（Ｐ＜０．０１）。Ⅲ组水貂的回肠胰蛋白酶活性显著高于Ⅵ组（Ｐ＜０．０５）,与其他各组差异不显著（Ｐ＞０．０５）。Ⅵ组水貂的十二指肠胰脂肪酶活性显著或极显著高于Ⅱ、Ⅳ和Ⅴ组（Ｐ＜０．０５或 Ｐ＜０．０１）,空肠胰脂肪酶活性显著或极显著高于Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ组（Ｐ＜０．０５或 Ｐ＜０．０１）。Ⅰ、Ⅲ和Ⅴ组水貂的回肠胰脂肪酶活性极显著高于Ⅱ和Ⅵ组（Ｐ＜０．０１）,与Ⅳ组差异不显著（Ｐ＞０．０５）。饲粮蛋白质水平为 ３６％时,水貂空肠绒毛高度、隐窝深度、绒毛高度与隐窝深度比值、黏膜厚度以及肠壁厚度均处在较高水平。由此得出,饲粮蛋白质水平可影响冬毛期水貂胃肠道消化酶活性,并可调节其空肠形态结构。     

希腊式酸奶的研制

研制希腊式酸奶,确定希腊式酸奶的最佳配方为鲜牛奶40%、白砂糖8%、脱脂奶粉：WPC为80：20、酪蛋白酸钠2‰、菌种为YO-MIXTM495;同时确定了制作希腊式酸奶的最佳水合条件为50℃、60min,发酵终点pH值控制在4.65。     

Acute phase proteins in cattle and swine: A review

The major acute phase proteins (APPs) in cattle are haptoglobin (Hp) and serum amyloid A (SAA), and in swine, are Hp, SAA, C-reactive protein (CRP), and Pig major acute phase protein (Pig-MAP). Many methodologic assays are presently available to measure these parameters, which are still being improved to increase their specificity, sensitivity, user-friendliness, and economic availability. In cattle, the main applications are the diagnosis and monitoring of frequent diseases such as mastitis and metritis in dairy cows and respiratory problems in young calves. In pigs, APPs are useful in the control of bacterial and viral infections, and they may be used at the slaughterhouse to monitor subclinical pathologies and improve food safety. The utility of APP in animal production must not be forgotten; optimization of protocols to improve performance, welfare, and nutrition may benefit from the use of APPs. Other sample types besides serum or plasma have potential uses; APP determination in milk is a powerful tool in the control of mastitis, saliva is a non-invasive sample type, and meat juice is easily obtained at the slaughterhouse. Increasing our knowledge of reference intervals and the influence of variables such as age, breed, sex, and the season is important. Finally, worldwide harmonization and standardization of analytical procedures will help to expand the use of APPs.     

Evaluation of a single dose versus a divided dose regimen of danofloxacin in treatment of Actinobacillus pleuropneumoniae infection in pigs

A single versus a divided dose regimen of danofloxacin was evaluated in treatment of porcine Actinobacillus pleuropneumoniae infection using clinical observations combined with biochemical infection markers: C-reactive protein, zinc and ascorbic acid. Twenty hours after experimental infection, the 18 pigs received danofloxacin intravenously as a single dose of 2.5mg/kg or four doses of 0.6 mg/kg administered at 24h intervals. These dosage regimens resulted in similar AUCs of the plasma danofloxacin vs time curve. The maximum concentration was 3.5-fold higher using the single dose regimen, while the time with concentrations above the MIC was 2.5-fold longer using the fractionated regimen. Using the single dose regimen, temperature was normalised 32 h post-infection. In contrast, normalisation was delayed until 44 h post-infection using four low doses and a relapse with elevated temperatures at 52 and 68 h was observed. No other significant differences between the treatments were found, neither regarding clinical, haematological nor biochemical observations. The use of the more convenient single dose regimen was appropriate, as it was at least equivalent to the fractionated regimen.     

低聚糖对雏鸡大肠粪便NH3-N含量及肠道菌群的影响

试验选取1日龄罗曼蛋公雏250只，随机分为5组，试验组饲料或饮水中分别添加海藻糖(500mg／kg)、海藻胶(600mg／kg)、大豆寡糖(500mg／kg)、圆葱寡糖(500mg/d)。结果表明，四种低聚糖均可促进肠道乳酸菌及双歧杆菌的生长繁殖，对魏氏梭菌的生长呈现抑制作用。海藻糖、海藻胶对降低大肠粪便NH3—N含量效果不显著(P＞0．05)，而大豆寡糖效果显著(P＜0．05)，圆葱寡糖效果极显著(P＜0.01)。     

Effects of Feeding Different Dietary Crude Protein and Digestible Energy to Yili Mares during Late Lactation Stage on Growth Development and Blood Biochemical Parameters of Foals

In this paper, thirty Yili mares aged 11 to 12 years with weighing 410 kg ± 30 kg, parity of 4 to 5 in third lactation month were selected, and were randomly divided into six groups, each group with five repeats.Two factors quadratic regression orthogonal rotational combination design was adopted to evaluate the effects of feeding different dietary CP and DE to mares on growth development and blood biochemical parameters of foals.Daily DE and CP feeding levels of the 6 groups were 92 MJ/d and 1.20 kg/d, 125 MJ/d and 1.20 kg/d, 92 MJ/d and 1.55 kg/d, 107 MJ/d and 1.35 kg/d, 125 MJ/d and 1.44 kg/d, 115 MJ/d and 1.55 kg/d, respectively.The results showed that body weight, body height, chest circumference and cannon circumference of foals of four and five months were not significant affected by maternal protein and energy intake (P>0.05).As compared with ration contained high DE (125 MJ/d) and low CP (1.20 kg/d), feeding with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) improved body length of foals of four and five months significantly (P<0.05).From the blood biochemical parameters, such as total protein, albumin, triglyceride, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, urea nitrogen, creatinine, alanine aminotransferase, there were no significant differences among all groups (P>0.05).Mares fed with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) obviously improved the aspartate aminotransferase and uric acid level.In summary, Yili mares during lactation late stage fed with 1.35 kg/d CP and 107 MJ/d DE daily could significantly improve body length development of foals, and foals nitrogen metabolism was altered by maternal dietary nutrition through increasing blood aspartate aminotransferase activity.     

Bioinformatics Analysis of Bone Morphogenetic Protein 4 (BMP4) of Sheep

The assay was aimed to explore the biological characteristics of bone morphogenetic protein 4 (BMP4) of sheep,NCBI,DNAMAN DNAStar,TMHMM Server v.2.0,PsortⅡ,SignalP various bioinformatical softwares were used to speculate the physical and chemical properties,hydrophobic property,phosphorylation site,conservative structure domain,protein secondary structure of BMP4 protein.Also,the three-dimensional structure was forecasted with the SWISS-MODEL Workspace software.The results indicated that the BMP4 of sheep had high homologies with the BMP4 of various species.The encoded protein was a hydrophilic protein which was unstable.There was no transmembrane regions and it was likely to be located in the nucleus.What was more,there was signal peptide and eighteen phosphorylation sites.Through the forecast of functional domains,the protein had two functional domains,including the transforming growth factor-beta (TGF-beta) superfamily and TGF-beta propeptide superfamily.The result was consistent with the function of BMP4 gene family,it also demonstrated that BMP4 was a growth factor and it had the function of signal transduction.The amino acid homology between the predicted 3D structure of protein and template 3bmp.1.A was 88.29%.The bioinformatics analysis of BMP4 gene could provide reference for the further study in practice.     

Influence of race and crossbreeding on casein micelles size

Casein (CN) micelles are colloidal aggregates of protein dispersed in milk, the importance of which in the dairy industry is related to functionality and yield in dairy products. The objective of this work was to investigate the correlation of milk CN micelles diameter from Holstein and Zebu crossbreds with milk composition (protein, fat, lactose, total and nonfat solids and milk urea nitrogen), somatic cell count (SCC), age, lactation stage and production. Average casein micelles diameters of milk samples obtained from 200 cows were measured using photon correlation spectroscopy and multiple regression analysis was used to find relationship between variables. CN micelle diameter, SCC and nonfat solids were different between animals with different Holstein crossbreed ratios, which suggests influence of genetic factors, mammary gland health and milk composition. Overall, results indicate the potential use of CN micelle diameter as a tool to select animals to produce milk more suitable to cheese production.