实验用小型猪信息管理系统开发

为满足小型猪培育数字化管理需要开发的"中国实验用小型猪信息管理系统"由育种管理、饲养管理、群体监测和操作规程等14个模块组成,其中最具特色的部分是育种管理和群体监测模块。为确保数据库质量,育种管理模块创建了以级联表和单选框勾选、个体号和群体类型自动生成的个体信息录入方式;根据近交系和封闭群建系规则创建了自动配种模块,基于系谱生成配种计划表,结合发情记录、产仔记录等实现繁殖管理数字化。群体监测模块由生物学特性、微生物和寄生虫质量控制、遗传质量控制三个部分组成,是建立高品质实验用小型猪群体的信息保障。     

宁乡花猪原代和第一代血液生理生化指标比较

目的比较原代宁乡花猪与其第一代仔猪血液生理生化指标的变化。方法常规方法测定宁乡花猪F0代16头,F1代16头血液18项生理指标和13项生化指标。结果F0代与F1代部分血液生理指标如白细胞（WBC）、红细胞（RBC）、血红蛋白（HGB）、红细胞积压（HCT）等13项有显著性差异（P〈0．05）;部分血液生化指标如总胆红素（TBIL）,尿素氮（BUN）,总蛋白（TP）,白／球蛋白（ALB／GLB）,葡萄糖（GLU）,有显著性差异（P〈0．05）;其他指标均无统计学差异。结论2代宁乡花猪之间部分血液生理生化指标有显著性差异。得到的数据可以为实验动物化研究提供相应的基础生理参考数据。     

宁乡猪皮下脂肪与肌内脂肪组织转录组差异分析

为了比较分析宁乡猪皮下脂肪(subcutaneous fat,SAF)和肌内脂肪(intramuscular fat,IMF)组织脂肪沉积的分子机制,本文利用RNA-seq技术鉴定和分析宁乡猪SAF和IMF组织中差异基因表达谱。选取6头250日龄健康状况良好、种内个体体重相近(约85 kg)的雄性宁乡猪为实验材料,采集SAF与IMF组织样品。通过对两个脂肪组织转录组测序并进行GO(Gene Ontology)功能注释及KEGG(Kyoto Encyclopedia of Genes and Genomes)信号通路富集分析,得到与脂肪沉积和脂质代谢相关的差异基因。为验证测序数据结果的可靠性,本研究随机选取6个差异基因进行qRT-PCR验证。结果表明,宁乡猪SAF和IMF组织中有2406个基因差异表达,其中1422个基因表达上调,984个基因表达下调。通过GO功能注释分析发现,差异表达基因主要通过参与类固醇生物合成、不饱和脂肪酸的生物合成、甘油磷脂代谢及自噬途径等与脂质代谢相关途径,调控宁乡猪SAF和IMF的沉积。KEGG通路富集结果显示,差异基因主要富集在脂质结合、脂肪酸代谢过程、甘油...     

国内外小型猪实验动物化研究

生命科学研究和生物医药产业的迅猛发展对实验动物,尤其是动物模型提出了越来越高的要求。猪在心血管系统、消化系统、皮肤结构、骨骼发育和营养代谢等方面与人极其相似,因此一直是医学研究的重要动物模型。更是人类异种器官、组织、细胞移植的首选供体。详细介绍了国内外小型猪实验动物化研究进展。     

东方田鼠在生物医学研究中的应用及展望

东方田鼠在分类、分布、形态、生态、实验室饲养、部分生物学特征、遗传学、微生物学和寄生虫学等方面的研究取得了一系列进展。近年来,东方田鼠开发为日本血吸虫病、糖尿病和自发性卵巢癌实验动物模型新品系。应用这些模型可对日本血吸虫病、糖尿病和自发性卵巢癌的病理学改变、发病机制、药物治疗和疫苗开发等多方面进行研究。本文就东方田鼠在生物医学研究中的应用研究现状作一综述。     

猪在生物医学研究中的应用

Susan Walton指出:几乎任何生物模型都可直接应用于人类的研究。但现在多侧重于哺乳动物模型〔1〕。近年来的研究表明,猪的许多解剖学、生理学特征与人相似,且猪的体型大小适度,性格温和,易于驯服,饱食后喜睡,可允许反复采样并进行各种手术,也容易选育成特殊用途的品种,故用它作为复制人类比较医学研究的动物模型,已广泛应用于生物医学研究中,有着广阔的前景。一、猪的部分解剖结构、生理生化指标接近人类家猪是由野猪驯化、培育而成,属偶蹄目野猪科。它既是杂食兽,又是甜食动物,味蕾能感觉甜味,其胃腺分布在整个胃壁上,最接近人类。猪与人…     

从大肠杆菌中表达、纯化宁乡猪N-乙酰谷氨酸合成酶并制备其多克隆抗体

N-乙酰谷氨酸合成酶催化生成的N-乙酰谷氨酸(NAGS)对于哺乳动物尿素循环第一个酶—氨基甲酰磷酸合成酶I变象异构激活是必需的。N-乙酰谷氨酸合成酶定位于肝脏和小肠线粒体基质中,通过提供N-乙酰谷氨酸调节氨基甲酰磷酸合成酶I的活性来调节尿素合成。我们用RT-PCR方法从宁乡猪肝脏中扩增了N-乙酰谷氨酸合成酶的开发阅读框,并将此基因连接到原核表达载体上,构建了pET-NAGS质粒。将重组质粒转化到Ec.oliBL21(DE3),在IPTG诱导下表达His-NAGS融合蛋白。通过SDS-PAGE,得出NAGS分子量约为40kDa。一步亲和层析纯化后,我们将纯化后的NAGS蛋白注射到新西兰大白兔中制备多克隆抗体。通过免疫组化和免疫印迹测试抗体,结果表明此抗体有较好的抗原性和特异性。据我们所知,这是第一次在大肠杆菌中表达来源于宁乡猪的NAGS。     

遗传修饰猪模型在生物医学及农业领域研究进展及应用

猪在解剖结构、代谢、生理生化等特征方面比啮齿类动物更接近人类,因此在模拟某些人类疾病以及提供异种移植器官等方面具有其他动物不可替代的优势,是理想的人类疾病动物模型和异种器官的供体。另外,猪作为我国畜牧业最重要的物种之一,猪的品种改良、疫病防控以及动物福利等问题都与人民生活息息相关。本文主要介绍了遗传修饰猪模型在分子育种、人类疾病模型以及异种器官移植领域的研究进展及未来应用前景,希望增进相关领域研究人员对基因编辑等前沿技术的了解,理解遗传修饰猪模型在生命科学研究中的重要意义。     

Development of a real‐time PCR method for Firmicutes and Bacteroidetes in faeces and its application to quantify intestinal population of obese and lean pigs

Aims: To investigate whether the relative abundance of the Bacteroidetes and Firmicutes divisions in pigs is different between obese and lean animals. Methods and Results: Group‐specific primers were designed to target the 16S rRNA genes of Bacteroidetes and Firmicutes present in the gut. After the validation of their specificity, these primers were used in the real‐time PCR quantification of all Bacteria, Firmicutes division, Bacteroidetes division and Bacteroides spp. in the faecal samples of obese and lean pigs from Banna mini‐pig inbred line. The obese pigs had a ～61% fewer percentage (based on all Bacteria) of Bacteroidetes division (P = 0·033) and a ～56% fewer proportion of Bacteroides spp. (P = 0·047) than the lean pigs. The proportions of both Bacteroidetes and Bacteroides had a negative correlation (P < 0·01) with the body weight. Conclusion: The results suggested that the fat storage might affect the proportion of Bacteroidetes division in the gut. Significance and Impact of the Study: The real‐time PCR assays developed for Firmicutes and Bacteroidetes will be useful for investigating the composition of gut microbiota.     

Histidine maintenance requirement and efficiency of its utilization in young pigs

Abstract

An experiment was conducted in young pigs (initial BW 10.1 kg) to estimate the maintenance requirement for histidine and its efficiency of utilization for protein accretion using a comparative slaughter technique. Three groups of six pigs each were fed a purified diet supplying 0, 14 or 56 mg histidine per kg BW^0.75. Following 21 d of feeding, pigs were killed for whole body compositional analysis. A representative group of six pigs was killed at the beginning of the experiment. Retention of histidine and total N were the main criteria of response. Histidine retention (R² = 0.73) and N retention (R² = 0.78) were linear functions of histidine intake (p < 0.001). Histidine requirement for zero histidine retention was 15.5 mg/kg BW^0.75, whereas histidine required for zero N retention was 4.1 mg/kg BW^0.75. At zero histidine retention, the pigs retained daily 82 mg N/kg BW^0.75, presumably due to the degradation of histidine-rich compounds such as haemoglobin and/or carnosine. The slope of the regression line relating histidine retention to N retention indicated that 105 mg of histidine was deposited per gram of total N which was considerably less than the estimated histidine concentration in body protein (179 mg/g N). Based on the slopes of regression equations for histidine and N retention, marginal efficiency of histidine utilization was calculated to be 0.94 and 1.34, respectively.     

The application of stable isotopes in biomedical research

Many of the ways in which isotopes are used in biomedical research are reviewed. The use of stable isotopes in stable isotope dilution assays, for metabolite identification and in pharmacokinetic studies, is discussed and relevant examples are given to illustrate the various points made. Isotope effects and their implications for future drug design are considered. Some of the toxicity problems associated with the use of stable isotopes are also considered. Finally, a brief subjective view of possible future advances is made.     

Molecular characterization of the human ABO blood group orthologus system in pigs

The selection and use of animals with blood group 0 in the process of transplanting pig organs or tissues into humans can positively contribute to the control of acute immune rejection due to differences in blood groups. Exon-specific PCRs for the porcine blood group A transferase gene against genomic DNA from either blood group A or 0 animals resulted in the amplification failure of the A0 blood group gene exon 8 from blood group 0 animals. To characterize the genetic abnormality in the genome of blood group 0 animals, we screened bacterial artificial chromosome (BAC) clones from a Korean native pig BAC library which had the blood group 0 allele, and carried out shotgun sequencing. The analysis showed that the 0 allele has a large deletion between exon 7 of the A0 blood group gene and the neighbouring SURF6. We also showed that the ABO blood group antigens in humans and the A0 blood group antigens in pigs are coded by mutations within the orthologous glycosyltransferase gene. In addition, we developed a multiplex genotyping method for the porcine A0 blood group gene.     

Validation of ERIC PCR as a tool in epidemiologic research of Salmonella in slaughter pigs

The purpose of this study was to test a protocol for a standardized ERIC PCR for its capability of genotyping Salmonella, isolated from pigs and their environment, in an epidemiologic approach. To test repeatability, four different Salmonella isolates were subjected to PCR three times. Furthermore, it was tested if the profiles on gel differed when a higher annealing temperature was used. Four Salmonella isolates were subjected to four different annealing temperatures (36, 40, 48 and 55°C). Moreover it was tested if the differentiation of Salmonella isolates, based on the genotypes, differed when a higher annealing temperature was used. Eight Salmonella isolates were tested at normal (36°C) and high (55°C) annealing temperatures. The results showed that this standardized ERIC PCR protocol was an efficient tool for typing many Salmonella isolates within a short period of time. The profiles were repeatable within one PCR reaction, but some profiles differed when they were compared between reactions. A higher annealing temperature resulted in profiles that contained more or fewer bands. The differentiation between isolates, when comparing profiles, remained the same. It was concluded that the standardized ERIC PCR protocol is useful for genotyping Salmonella. Received 26 January 1998/ Accepted in revised form 3 August 1998     

扫描探针显微镜及其在生物医学研究中的应用

本文介绍了一类可以从原子水平到微米尺寸观察物质结构的三维成像工具——扫描探针显微镜(SPM),重点介绍了扫描隧道显微镜(STM)和原子力显微镜(AFM)的基本原理,以及SPM在细胞生物学、核酸和小分子成像等生物医学研究领域的一些应用。SPM不久将可能成为大多数生命科学实验室的一项重要技术。     

特殊理化微环境的构建及其在生物医学领域的应用

生物医学领域中的微环境是指在实质细胞周围参与组织细胞生理及病理过程的微小特殊区域,它对于调控细胞的运动、增殖、分化、分泌及代谢功能具有重要作用。本文在介绍体内微环境及其组成的基础上,重点阐述了体外微环境构建技术及其在生物医学领域的应用,展望了微环境生理作用机制研究的发展趋势。     

Effects of phosphorus intake on phosphorus flow in growing pigs: application and comparison of two models

A comparison of the models of Vitti et al. (2000, J. Anim. Sci. 78, 2706-2712) and Fernández (1995c, Livest. Prod. Sci. 41, 255-261) was carried out using two data sets on growing pigs as input. The two models compared were based on similar basic principles, although their aims and calculations differed. The Vitti model employs the rate:state formalism and describes phosphorus (P) flow between four pools representing P content in gut, blood, bone and soft tissue in growing goats. The Fernández model describes flow and fractional recirculation between P pools in gut, blood and bone in growing pigs. The results from both models showed similar trends for P absorption from gut to blood and net retention in bone with increasing P intake, with the exception of the 65 kg results from Date Set 2 calculated using the Fernández model. Endogenous loss from blood back to gut increased faster with increasing P intake in the Fernández than in the Vitti model for Data Set 1. However, for Data Set 2, endogenous loss increased with increasing P intake using the Vitti model, but decreased when calculated using the Fernández model. Incorporation of P into bone was not influenced by intake in the Fernández model, while in the Vitti model there was an increasing trend. The Fernández model produced a pattern of decreasing resorption in bone with increasing P intake, with one of the data sets, which was not observed when using the Vitti model. The pigs maintained their P homeostasis in blood by regulation of P excretion in urine.