一个RNAi载体上反向重复片段的测序策略

相邻的反向重复DNA片段有形成单链内二级结构的倾向,属于一种测序困难的DNA模板。解决RNAi载体插入的反向重复片段的测序问题,为该类载体正确性的测序验证奠定基础。采用常规分子克隆方法构建表达小麦TaATG2串联反向重复片段的RNAi载体,设计2种策略对经菌落PCR初步鉴定的载体进行测序验证:一种是以完整的载体质粒为模板进行测序;另一种是先对载体进行酶切处理,切除反向重复片段中的一个后对保留另一个片段的线性载体进行测序。结果表明,第一种测序策略受到串联反向重复片段形成的单链内部二级结构的影响,测序信号在反向重复片段处出现衰减或乱峰,无法读取序列。第二种测序策略排除了2个反向重复片段之间的干扰,保留在载体上的片段测序信号清晰,序列准确。采用酶切切除一个片段后进行测序的方法,经过2次酶切和2次测序可以有效地对载体上的2个反向重复片段分别进行序列测定,进而确认构建载体的正确性。     

Epigenetic Regulation of Elf5 Is Associated with Epithelial-Mesenchymal Transition in Urothelial Cancer

E74-like factor 5 (Elf5) has been associated with tumor suppression in breast cancer. However, its role in urothelial cancer (UC) is completely unknown. Immunohistochemistry (IHC) and methylation specific PCR (MSP) were done to detect Elf5 expression level and its promoter methylation. Results revealed that low expression of Elf5 on protein and mRNA levels were associated with tumor progression, early relapse and poor survival. In vitro, down-regulation of Elf5 can increase epithelial-mesenchymal transition (EMT). Aberrant Elf5 methylation was identified as major mechanism for Elf5 gene silence. Accordingly, restoration of Elf5 by infection or demethylating treatment effectively reversed EMT processes. In conclusion, we identified Elf5 as a novel biomarker of UC on several biological levels and established a causative link between Elf5 and EMT in UC.     

Cell sheet technology: a promising strategy in regenerative medicine

Regenerative medicine is a burgeoning field that is important to combat challenging diseases and functional impairments. Compared with traditional cell therapies with evident shortcomings (e.g., cell suspension injection or tissue engineering with scaffolds), scaffold-free cell sheet technology enables transplanted cells to be grafted and fully maintain their viability on target sites. Clinical and experimental studies have advanced the application of cell sheet technology to numerous tissues and organs (e.g., liver, cornea and bone). However, previous reviews have failed to discuss vital aspects of this rapidly developing technology, and many new challenges are gradually emerging. This review aims to provide a comprehensive introduction to cell sheet technology from cell selection to the ultimate applications of cell sheets, and challenges and future visions are also described.     

图形视网膜电图的空间和时间调谐特性——FFT分析

用快速傅里叶转换(FFT)技术分析了图形视网膜电图(PERG)的空间和时间调谐特性。PERG 的二次谐波在较高的空间频率(>0.46周/度)逐渐下降,与 PERG振幅的变化相似,但显示明显的低空间频率衰减。空间和时间调谐特性存在一定的相关。引起最大二次谐波振幅的最佳时间频率,在低空间频率时(<0.23周/度)由低频(≤3.91Hz)移至 7.81Hz。     

豚鼠心肌α-肾上腺素受体反应的可能机制——增强细胞外钙离子内流

同时记录豚鼠心室肌细胞动作电位、零期最大去极化速率和心肌条的收缩幅度,观察了α-肾上腺素受体激动剂或增加细胞外钙离子浓度([Ca~(2 )]_0)的作用,以及在肾上腺素受体或钙通道阻断剂存在下这些作用的变化,试图分析α-受体与钙通道的可能关系。我们看到甲氧胺与增加[Ca~(2 )]_0都可使动作电位峰值增加、平台期电位高抬、收缩幅度增高,甲氧胺的效应可被戊脉胺阻断,而增加[Ca~(2 )]_0的效应可被酚妥拉明阻断。酚妥拉明同戊脉安两者的效应有近似之处,心得安对甲氧胺和增加[Ca~(2 )]_0效应无显著影响。由此认为:心肌α-肾上腺素受体的电生理效应与缓慢正性变力作用很可能是通过激活钙通道,增加细胞外钙离子的慢内流来实现的。