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。     

鳢肠对棉铃虫生长发育的影响

通过饲料混毒法测定鳢肠对棉铃虫生长发育的影响。结果表明,随饲料中鳢肠比例的加大和棉铃虫幼虫取食时间的延长,鳢肠对棉铃虫幼虫的生长发育具有明显的抑制作用。幼虫取食鳢肠比例高的饲料后,棉铃虫蛹质量显著降低,但羽化率和单雌日均产卵量与对照处理差异不显著;给棉铃虫幼虫饲喂鳢肠明显延长了幼虫和成虫历期,但对蛹历期的影响较小。     

黑茶茶多酚类物质的提取及其抗氧化性能研究

以湖南安化天尖、茯砖及广西六堡3种黑茶为考察对象,研究黑茶中茶多酚的提取工艺以及抗氧化性能。针对超声波不同提取条件,采用正交试验,确定黑茶中茶多酚的最佳提取工艺条件;采用清除二苯代苦味酰自由基(DPPH)的能力和邻苯三酚自氧化体系来评价其抗氧化活性。结果表明:采用超声波热水复合浸提法提取黑茶中茶多酚的最佳提取条件为超声波提取功率250 W,提取固液比1∶25,提取温度55℃,提取时间45min,茯砖黑茶中茶多酚的提取率5.21%;同时测得茯砖黑茶具有较好的抗氧化能力,对DPPH自由基有较强的清除能力,但与维生素C(VC)相比,其抗氧化能力比VC的能力弱;其清除超氧阴离子自由基能力的IC50为0.507μg/mL。     

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。     

贵州蕨类植物模式标本种的整理研究

经查阅文献,搜集并整理得到模式标本采自贵州的蕨类植物19科、46属、126种.模式产地涉及除六盘水地区外的贵州各地区,黔南地区51种＞安顺地区18种＞贵阳地区17种＞黔西南地区7种、铜仁地区7种、遵义地区7种＞毕节地区4种＞黔东南地区1种,产地不祥的14种.种数较多的科依次是鳞毛蕨科40种＞金星蕨科16种＞蹄盖蕨科15种＞水龙骨科10种.种数较多的属是耳蕨属11种,其次是贯众属10种.生长环境具有多样性,生长种数较多的生境依次是林下生长种类94种＞石上或石隙生长种类62种＞溪沟边生长种类41种＞阴湿环境生长种类30种＞河谷或峡谷生长种类23种.     

蓖麻环核苷酸门控离子通道RcCNGC2克隆与盐胁迫下表达分析

为探讨RcCNGC2在蓖麻耐盐中的作用,以通蓖5号叶片为材料,克隆获得环核苷酸门控离子通道RcCNGC2完整编码区序列(CDS),并对该序列进行序列比对、蛋白结构分析,构建多元表达载体,分析了在盐胁迫下蓖麻RcCNGC2基因根、茎、叶组织中6个时间点的表达量变化。结果显示,RcCNGC2 CDS长度为2 148 bp,编码715个氨基酸,蛋白分子量为34.15 ku,等电点为9.96,存在5个跨膜结构域。氨基酸序列一致性分析表明,RcCNGC2与橡胶树一致性最高,达89.15%。qRT-PCR分析结果显示,NaCl处理后RcCNGC2表达量在根中上调且差异显著,在茎、叶中随NaCl处理时间延长而显著减少。综上,RcCNGC2在蓖麻受到盐胁迫时起重要信号传导作用。     

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。