Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Immunological and pathological investigations in equine experimental uveitis

Background

The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist.

Objectives

The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings.

Methods

Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-inflamed eyes taken from 6 horses in slaughter house. After centrifugation of the two eye media, smears were prepared and cytological investigation was performed. Tissue specimens were taken from all eye globes and were submitted to histopathological investigation.

Results

CRALBP and IL-17 concentrations were significantly elevated in eye media of horses with experimental uveitis in comparison with controls. Cytological and histopathological findings corresponded to the changes characteristic of chronic immune-mediated inflammation with mononuclear cell infiltration of uvea, choroid, retina, and eye media as well as severe retinal destruction.

Conclusions

Our study demonstrated the involvement of the retinal autoantigen CRALBP as well as IL-17 in the pathogenesis of experimental uveitis in horses. These findings suggests that this experimental uveitis in horses may serve as a suitable animal model for investigation of IL-17- mediated immune response during spontaneous autoimmune uveitis in horses as well as in humans.

     

贮藏温度对采后“金艳”猕猴桃品质和后熟的影响

以"金艳"猕猴桃为试材,采用不同温度贮藏的方法,研究了温度对果实品质和呼吸强度的影响。结果表明:1℃贮藏可以很好的保持果实品质,呼吸强度峰值出现晚,保持了较高的果肉硬度;5℃和10℃贮藏能够促进果实的后熟,加速呼吸强度峰值的到来、果实硬度的下降和可溶性固形物含量的上升;但35℃贮藏的果实7d后会失去商品价值,发生腐败;16℃贮藏果实后熟品质不佳。在整个贮藏过程中,果实干物质含量在上述温度条件下贮藏变化都不明显,但在一定范围内,干物质含量会随着温度的上升而有所增加。     

缺素胁迫对软枣猕猴桃幼苗生长的影响

以软枣猕猴桃幼苗为试材,研究了缺铁(Fe)、镁(Mg)、磷(P)、钙(Ca)、钾(K)、氮(N)6种元素对其幼苗生长及生理指标的影响。结果表明:不同缺素处理对其幼苗的生长发育及生理指标均能产生不同程度的影响,其缺钙(-Ca)、钾(-K)、氮(-N)对软枣猕猴桃幼苗根长、株质量、株高影响显著;游离脯氨酸(Pro)含量、丙二醛(MDA)含量、过氧化物酶(POD)活性及超氧化物歧化酶(SOD)活性均明显增加。     

三种地被菊种子萌发及幼苗生长的化感效应比较

以天人菊、金鸡菊和蛇鞭菊为试材,比较两两混种与单种在种子萌发和幼苗生长阶段相互间的化感效应强度。结果表明:两两混种下,金鸡菊、蛇鞭菊对天人菊生长起促进作用,化感效应强度为金鸡菊蛇鞭菊;天人菊、蛇鞭菊对金鸡菊生长起抑制作用,化感效应强度为天人菊蛇鞭菊;金鸡菊、天人菊对蛇鞭菊生长起抑制作用,化感效应强度为金鸡菊天人菊。     

野生老芒麦种质资源鉴定与基部叶鞘绒毛变异分析

为更好地利用老芒麦(Elymus sibiricus L.)野生资源,本研究从我国青藏高原、西北、华北、东北地区以及国外部分地区共采集了1 723份野生披碱草属种质资源,通过表型特征观测和流式细胞仪检测,共鉴定出了990份野生老芒麦种质,246份垂穗披碱草种质,并测得老芒麦的DNA含量在5.86～7.30 Gb之间,平均DNA含量为6.66 Gb;垂穗披碱草材料的DNA含量在9.50～10.36 Gb之间,平均DNA含量为9.97 Gb。研究结果显示,在西北、华北以及东北地区的部分野生老芒麦材料在苗期基部叶鞘出现了绒毛,分析发现该表型出现的概率与经度和纬度呈极显著正相关关系(P<0.01),与海拔、年年均气温以及年平均降雨量呈极显著负相关关系(P<0.01)。表明老芒麦苗期基部叶鞘绒毛的有无与环境因子之间存在着必然的联系,可能是老芒麦种质应对不同环境类型的适应性机制之一。本研究为老芒麦的形态鉴定提供了新的依据,为老芒麦种质资源挖掘和育种应用提供了材料基础。     

植物基因启动子的克隆方法及其应用

植物基因的表达调控已成为分子生物学研究热点,启动子是基因表达调控的重要顺式元件,启动子的克隆对于研究基因表达调控、构建基因工程载体、表达目的蛋白有着重要的意义。启动子克隆的方法很多,从常用的启动子陷阱技术筛选启动子到PCR方法的应用,此后相继问世的一些基于PCR的克隆启动子技术,如载体锚定PCR、反向PCR、接头PCR、交错热不对称PCR等,为克隆启动子提供了更可靠,更合理的方法。本文着重介绍了几种植物基因启动子的克隆方法,分析了它们的优缺点,并展望了今后的研究前景。