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.     

紫花苜蓿细胞质雄性不育恢复基因的初步定位

以紫花苜蓿(Medicago sativa)不育系MS-GN-1A为母本,恢复系MS178为父本组合构建BSA分离群体,获得的F1代均表现为雄性可育,在大田种植了221株F2代群体单株,盛花期将花粉颗粒染色后,显微镜下观察统计出,不育的F2代株数为57,可育F2代株数164,并没有观察到半不育植株。将所有植株划分为可育和不育两组,并构建可育和不育DNA混池,混池DNA从可育和不育植株组DNA中各随机抽取20个样品,以此对恢复基因定位。随机挑选160对已知的四倍体苜蓿SSR引物扩增基因池DNA,获得2个具有多态性的分子标记,分别是Mt2c12、AW166,初步定位Rf基因在类群Composite5上。将类群Composite5上的所有引物进行合成,进一步进行引物筛选,最终获得4个具有多态性的标记BI68、Mt2c12、BG267和AW776153,遗传距离分别为19.0、20.9、44.6和72.1cM。     

一起未经执业兽医注册从事诊疗活动案引发的思考

2017年成都市办理的一起未经兽医执业注册而进行犬只声带切除的行政处罚案件,引发了笔者对动物诊疗机构监督执法中"动物诊疗"范畴界定、动物诊疗许可证申请对象探究、"一事不再罚"原则的适用、"责令停业诊疗活动"与"责令停产停业"区别等问题的思考。本文以法学理论和相关法律规定为基础,对成都市动物诊疗机构执法实践的疑点与难点进行分析,提出执法人员应更准确适用法律法规,并呼吁尽快建立全国执业兽医查询系统的建议。     

鸭圆环病毒检测方法研究进展

鸭圆环病毒病是由鸭圆环病毒感染引起的一种免疫抑制性疾病。本文综述了鸭圆环病毒血清学和病原学检测方法的研究进展,主要包括间接免疫荧光试验、酶联免疫吸附试验、核酸探针技术、普通PCR、限制性片段长度多态性聚合酶链反应、套式PCR、多重PCR、荧光定量PCR和环介导等温扩增技术等,并对各种方法的优缺点进行了汇总分析,以期为该病的诊断与防控工作提供参考。     

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。     

慈竹生长过程纤维素、木质素及灰分动态积累

以当年生不同生长高度的慈竹为研究对象,探讨慈竹生长过程中纤维素、木质素和灰分含量的积累规律。结果表明:慈竹不同生长高度纤维素含量顶、中部变化趋势均以对数方式增长;基部纤维素含量和茎秆纤维素总含量变化趋势都呈"S型"曲线;各部位木质素含量变化趋势总体呈倒"S型"曲线,且不同高度间的木质素含量的差异达显著水平;随着生长高度的增高,茎秆顶部、基部灰分含量以及茎秆灰分总含量总体呈倒"S型"曲线变化,中部灰分含量总体呈"S型"曲线变化。纤维素总含量、木质素总含量的积累分别集中在茎秆高度为150~800、30~800 cm这个生长时期;灰分总量在300~800 cm保持较高的水平。     

辽宁省农村土地流转的金融服务探索

随着经济全球化的发展和我国现代化产业结构的不断调整,粮食安全的支撑重要性越发明显,但农村目前的青壮年劳动力却面临主动或被动脱离土地的窘境。如何推动农村土地流转,提升土地利用率,增加农业生产效应,加强其相关金融服务是一项重要有效途径。通过研究,发现辽宁省农村土地流转的金融服务存在一系列的问题,并给出相应的政策建议。     

金安桥水电站初次蓄水对导流洞稳定性影响的计算分析

金安桥水电站下闸蓄水后，陡升的库水位将对导流洞结构稳定造成不利影响。为保障堵头封堵前的导流洞结构稳定性，运用三维弹塑性损伤有限元，在模拟进水口边坡及导流洞开挖支护的基础上，计算汛前下闸水位时的岩体渗流场，对导流洞围岩和衬砌在岩体渗流作用下的结构稳定性进行复核分析。分析结果表明，导流洞的围岩稳定性在蓄水后仍有保障，衬砌结构受力较小，现有环向配筋满足结构内力要求，但在进水口变截面和导流洞变截面部位沿水流方向的拉应力较大，使得衬砌出现开裂。进一步分析表明，上述区域的开裂范围有限，对整体衬砌结构影响较小。