Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative real-time PCR was performed to analyze gene expression. As results, expression levels of pluripotent markers were increased depending on spheroid size and the production of PGE₂ was increased in spheroid formation compared to that in monolayer. Ki-67 showed a remarkable increase in the spheroid formed with 2.0?×?10⁵ cells/drop as compared to that in the monolayer. Expression levels of angiogenesis-inducing factors such as VEGF, IL-6, IL-8, and IL-18 were significantly increased in spheroid formation compared to those in the monolayer. Expression levels of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1 were also significantly increased in spheroid formation compared to those in the monolayer. Expression levels of osteocyte-specific markers such as RUNX2, osteocalcin, and differentiation potential were also significantly increased in spheroid formation compared to those in the monolayer. Therefore, spheroid formation of eAD-MSCs through the hanging drop method can increases the expression of angiogenesis-inducing and bone morphogenesis-inducing factors under optimal culture conditions.     

Effect of feed intake level and dietary protein content on the body temperature of pigs housed under thermo neutral conditions

Feed intake and diet composition appear to affect the body temperature of pigs. Two trials were conducted to analyse the effect of feed intake level and dietary protein content on the intestinal temperature (IT) of pigs housed under thermo neutral conditions. Ten pigs (64.1 ± 1.3 kg initial body weight) fitted with an ileal cannula were used. A thermometer set to register the IT at 5‐min intervals was implanted into the ileum through the cannula. In both trials, the ambient temperature ranged from 19.1 to 21.6°C and the pigs were fed at 07:00 and 19:00 hr (same amount each time). In trial 1, the pigs were fed daily 1.2 or 1.8 kg of a wheat–soybean meal diet. The IT followed a similar pattern along a 24‐hr period regardless the feed intake level. The IT rapidly increased up to 0.61 and 0.74°C after the morning meal and up to 0.53 and 0.47°C after the evening meal in pigs fed 1.2 and 1.8 kg/d respectively. The postprandial IT was higher in pigs fed 1.8 kg after each meal (p < .05). In trial 2, pigs were fed daily 1.8 kg of a low (11%) or a high (22%) crude protein diet. The IT followed a similar pattern along the 24‐hr period regardless the dietary protein level. The postprandial IT did not differ between pigs fed the low protein or the high protein (p > .10). The IT rapidly increased up to 0.66 and 0.62°C after the morning meal in pigs fed the high‐ and low‐protein diet (p < .05), but there was no change after the evening meal (p > .10). In conclusion, the feed intake level affected the IT of pigs housed under TN conditions, but the dietary protein content had no effect.     

犊牛魏氏梭菌的分离鉴定及致病性研究

2017年1月,山东省某规模化奶牛场10～20日龄犊牛出现腹泻、拉血便、呼吸困难等症状,部分犊牛突然死亡。通过流行病学调查和尸体剖检,初步怀疑为魏氏梭菌感染。采集病料进行病理组织学检查、细菌分离鉴定、生化试验、16S及毒素基因检测等,证明疫情由A型魏氏梭菌所致。对死亡犊牛的肠内容物及分离菌株进行致病性研究,发现肠内容物及分离菌株具有较强的致病性。药敏试验结果显示,该菌对青霉素、头孢噻呋、恩诺沙星等表现极敏感。根据病原确诊情况及药敏试验结果,指导临床用药,及时控制了牛场疫情。     

陕西百年农业科技发展研究

笔者回顾了陕西百年来不同时期农业科技发展的历史背景与社会环境,分析了陕西农业科技发展的动因与本质、范围与对象、社会需求等内在属性特征;总结了陕西农业科技发展的主要成效,即构建了农业科研技术服务体系、培育推广了技术良种方法、发展了农业产业、形成了农业科技发展的政策人才环境、改变了农业传统观念与生产方式;归纳凝练出陕西农业科技发展的经验启示,即农业科技发展是解决农业诸多矛盾的必然要求、实现农业现代化的有效途径、构建农业安全的重要举措,同时农业科技发展必须以国家需要、人民需求为根本出发点。指出陕西农业科技发展中成果转化率不高、体制机制僵化、省级支持不够、基层人员结构不合理等问题,提出通过“科研面向需求”、设立省级基金、开展农技有偿服务、扩大农技部门人事权限、发挥各类市场主体作用来解决问题,推动全省农业科技持续稳步发展。     

北京妙峰山森林文化条件价值评估

【目的】评价妙峰山森林文化价值，对我国其他地区的森林文化价值评估亦有一定的借鉴意义，为全面开展中国森林文化价值量化评估探索路径。【方法】采用条件价值评估法，调研受访者对森林文化的认同程度，评估妙峰山森林文化价值，继而采用逐步回归法分析受访者支付意愿的影响因素。【结果】古树名木、寓言典故和神山传说是受访者接触程度最高的文化符号；受访者了解森林文化的方式呈现多元化，互联网和书籍是最主要的途径；森林采摘是受访者最乐于接受的森林实践活动；94%以上的受访者认同森林具有文化价值，认可程度较高；舒缓心情是受访者最为认同的福利(占比为82.32%)，其次为锻炼身体(占比为62.20%)和休闲娱乐(占比为45.12%)；妙峰山森林文化的人均支付意愿为100元人民币，最重要的影响因素是个人年收入，其次是对森林文化价值的认同程度和学历；对所支付费用的使用透明度不放心是部分受访者拒绝支付的最主要原因；大部分受访者(占比为53.05%)愿意将所支付的费用使用在妙峰山森林文化艺术创作方面，捐款(占比为33.77%)是受访者最愿意接受的支付方式。【结论】理解偏差、心理账户偏差、支付方式偏差、投标起点偏差等多种偏差都会造成条件价值评估结果的误差。即使本研究只是初步估值，本研究的方法与结论仍然可以为建设与保护森林文化的相关管理决策提供依据。另外，如何提升国民对森林文化的认知和参与程度以及对森林文化所带来福利的感受，是提升国民对森林文化的认可程度，乃至提升森林文化价值的关键。森林文化的内涵随着人类对森林功能认识的不断深入、人类与森林相互作用关系的变化而不断拓展，如何正确认识森林文化并科学计量森林文化的价值仍然是一个有活力的课题，值得继续深入研究。     

土耳其斯坦裂体吸虫23kDa蛋白基因的序列分析及抗原表位预测

为研究土耳其斯坦裂体吸虫23 kDa蛋白的生物学特性,以土耳其斯坦裂体吸虫cDNA为模版,利用PCR对23 kDa基因进行扩增,并将其克隆到T-easy载体后进行序列测定。利用生物信息学对其结构、抗原指数进行分析,并对其抗原表位进行预测。序列分析结果表明,该虫体的23 kDa基因长度为657 bp,A+T含量为57.38%,与曼氏血吸虫、埃及血吸虫和日本血吸虫的23 kDa基因的相似性分别为85.24%、83.71%和81.89%。蛋白二级结构分析表明,23 kDa蛋白经过4次跨膜,主要由6个α-螺旋、3个β-折叠、7个β转角和若干个无规则卷曲构成。抗原表位预测结果表明,该蛋白有3个B细胞抗原表位。综合分析土耳其斯坦裂体吸虫23 kDa蛋白是一种较好的抗原分子,是土耳其斯坦裂体吸虫疫苗的重要候选分子。     

苦瓜对8种蔬菜的化感作用

以8种海南主要栽培的蔬菜为受体,采用室内生测法研究苦瓜(Momordica charantiaL.)茎叶水浸提液对其化感作用。结果表明:苦瓜茎叶水浸提液对8种受体植物的种子萌发和幼苗生长均有一定的化感效应,且化感作用方式和强度在受体间及指标间存在显著差异;化感作用综合效应的平均值大小顺序为:辣椒黄瓜豇豆菜豆萝卜樱桃番茄胡萝卜番茄。试验结果为建立合理的栽培制度提供了理论和试验依据。     

气相色谱-串联质谱法测定柑橘中5种杀螨剂残留

建立了气相色谱-串联质谱法(GC-MS/MS)检测柑橘中5种杀螨剂残留分析方法。样品经乙腈提取、氯化钠盐析后,取有机相用Qu EChERS方法净化,利用GC-MS/MS在多反应监测(MRM)模式下进行测定,外标法定量。5种杀螨剂在0.005~0.5mg/L浓度范围内呈现良好的线性关系,相关系数r^2均>0.996,方法定量限为0.005 mg/kg。在0.005、0.01和0.1mg/kg 3种添加水平下,加标回收率为70.6%~111.3%,相对标准偏差(RSD)为1.07%~14.64%。本方法简便、重现性良好,可用于柑橘类水果样品中杀螨剂的日常检测。     

猪瘟病毒衣壳蛋白与宿主丝/苏氨酸蛋白激酶N1相互作用的鉴定

为筛选并鉴定与猪瘟病毒(CSFV)衣壳(C)蛋白相互作用的宿主蛋白,本研究采用酵母双杂交技术以CSFV C蛋白为诱饵从猪外周血单个核细胞(PBMC)c DNA表达文库中筛选与之相互作用的宿主蛋白,共筛选到6种蛋白,分别为ATP5B、SON3、PKN1、PCBP1、RPS20和IQGAP1,根据Gene Ontology分析结果,这些蛋白分别参与细胞的增殖和代谢等过程。本研究选取丝/苏氨酸蛋白激酶N1(PKN1)进行进一步验证,经酵母共转化试验、免疫共沉淀试验和GST pull-down试验证实,宿主PKN1蛋白与CSFV C蛋白之间存在特异性结合。本研究首次证明PKN1与CSFV蛋白之间的相互作用关系,为进一步研究PKN1蛋白在CSFV感染过程中发挥的作用奠定了基础。