本研究旨在克隆犏牛前列腺素D合成酶(prostagland D synthase,L-PGDS)基因序列,并检测其在不同组织及不同发育时期睾丸中的表达水平,从而为深入研究L-PGDS基因在犏牛睾丸发育过程中的作用机制提供依据。采集成年健康犏牛心脏、肝脏、脾...本研究旨在克隆犏牛前列腺素D合成酶(prostagland D synthase,L-PGDS)基因序列,并检测其在不同组织及不同发育时期睾丸中的表达水平,从而为深入研究L-PGDS基因在犏牛睾丸发育过程中的作用机制提供依据。采集成年健康犏牛心脏、肝脏、脾脏、肺脏、肾脏、大肠、小肠、胃、大脑及不同发育时期犏牛睾丸组织:胎牛期(5~6月)、幼年期(1~2岁)、青春期(2.5~4岁)、老年期(7~9岁),Trizol法提取各组织总RNA,利用RT-PCR技术扩增、克隆L-PGDS基因序列并利用相关生物学软件进行分析,利用实时荧光定量PCR检测犏牛各组织及4个不同发育阶段睾丸中L-PGDS基因mRNA的表达水平。结果显示,L-PGDS基因序列全长624 bp,包括完整的开放阅读框576 bp,编码191个氨基酸。同源性比对发现,犏牛与黄牛和绵羊的同源性最高,分别为99.28%和94.0%。系统进化树分析结果显示,犏牛与黄牛亲缘关系最近,其次是绵羊、马。L-PGDS蛋白为不稳定疏水蛋白,分子质量为21.229 ku,分子式为C953H1471N251O281S9,等电点为6.43,不稳定指数为47.25,不含跨膜区及信号肽,蛋白质二级结构预测显示α-螺旋、无规则卷曲、延伸链分别占25.65%、53.40%和20.94%。实时荧光定量PCR检测结果显示,L-PGDS基因在犏牛睾丸组织中特异性高表达,极显著高于其他组织(P<0.01);随着年龄增长L-PGDS基因mRNA在犏牛睾丸中呈先上升后下降趋势,其中在青春期相对表达量最高,极显著高于其他时期(P<0.01)。本研究结果为深入探究L-PGDS基因在犏牛睾丸发育过程中的作用机制提供了基础资料。展开更多
Textile-based technologies are considered as potential routes for the production of 3D porous architectures for tissue engineering( TE) applications. We describe the use of two polymers,namely polybutylene succinate( ...Textile-based technologies are considered as potential routes for the production of 3D porous architectures for tissue engineering( TE) applications. We describe the use of two polymers,namely polybutylene succinate( PBS) and silk fibroin(SF) to produce fiber-based finely tuned porous architectures by weft and warp knittings. The obtained knitted constructs are described in terms of their morphology, mechanical properties,swelling ability,degradation behaviour,and cytotoxicity. Each type of polymer fibers allows for the processing of a very reproducible intra-architectural scaffold geometry,with distinct characteristics in terms of the surface physicochemistry,mechanical performance,and degradation capability,which has an impact on the resulting cell behaviour at the surface of the respective biotextiles. Preliminary cytotoxicity screening shows that both materials can support cell adhesion and proliferation. Furthermore, different surface modifications were performed( acid /alkaline treatment, UV radiation,and plasma) for modulating cell behavior. An increase of cell-material interactions were observed,indicating the important role of materials surface in the first hours of culturing. Human adipose-derived stem cells( hASCs) became an emerging possibility for regenerative medicine and tissue replacement therapies. The potential of the recently developed silk-based biotextile structures to promote hASCs adhesion,proliferation,and differentiation is also evaluated. The obtained results validate the developed constructs as viable matrices for TE applications. Given the processing efficacy and versatility of the knitting technology, and the interesting structural and surface properties of the proposed polymer fibers,it is foreseen that our developed systems can be attractive for the functional engineering of tissues such as bone,skin,ligaments or cartilage and also for develop more complex systems for further industrialization of TE products.展开更多
文摘Textile-based technologies are considered as potential routes for the production of 3D porous architectures for tissue engineering( TE) applications. We describe the use of two polymers,namely polybutylene succinate( PBS) and silk fibroin(SF) to produce fiber-based finely tuned porous architectures by weft and warp knittings. The obtained knitted constructs are described in terms of their morphology, mechanical properties,swelling ability,degradation behaviour,and cytotoxicity. Each type of polymer fibers allows for the processing of a very reproducible intra-architectural scaffold geometry,with distinct characteristics in terms of the surface physicochemistry,mechanical performance,and degradation capability,which has an impact on the resulting cell behaviour at the surface of the respective biotextiles. Preliminary cytotoxicity screening shows that both materials can support cell adhesion and proliferation. Furthermore, different surface modifications were performed( acid /alkaline treatment, UV radiation,and plasma) for modulating cell behavior. An increase of cell-material interactions were observed,indicating the important role of materials surface in the first hours of culturing. Human adipose-derived stem cells( hASCs) became an emerging possibility for regenerative medicine and tissue replacement therapies. The potential of the recently developed silk-based biotextile structures to promote hASCs adhesion,proliferation,and differentiation is also evaluated. The obtained results validate the developed constructs as viable matrices for TE applications. Given the processing efficacy and versatility of the knitting technology, and the interesting structural and surface properties of the proposed polymer fibers,it is foreseen that our developed systems can be attractive for the functional engineering of tissues such as bone,skin,ligaments or cartilage and also for develop more complex systems for further industrialization of TE products.
