Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

Progressive cardiac fibrosis accelerates the development of heart failure. Here, we aimed to explore serum Wnt5a and Wnt11 levels in hypertension patients, the roles of Wnt5a and Wnt11 in cardiac fibrosis and potential mechanisms under pressure overload. The pressure overload mouse model was built by transverse aortic constriction (TAC). Cardiac fibrosis was analyzed by Masson’s staining. Serum Wnt5a or Wnt11 was elevated and associated with diastolic dysfunction in hypertension patients. TAC enhanced the expression and secretion of Wnt5a or Wnt11 from cardiomyocytes (CMs), cardiac fibroblasts (CFs), and cardiac microvascular endothelial cells (CMECs). Knockdown of Wnt5a and Wnt11 greatly improved cardiac fibrosis and function at 4 weeks after TAC. In vitro, shWnt5a or shWnt11 lentivirus transfection inhibited pro-fibrotic effects in CFs under mechanical stretch (MS). Similarly, conditional medium from stretched-CMs transfected with shWnt5a or shWnt11 lentivirus significantly suppressed the pro-fibrotic effects induced by conditional medium from stretched-CMs. These data suggested that CMs- or CFs-derived Wnt5a or Wnt11 showed a pro-fibrotic effect under pressure overload. In vitro, exogenous Wnt5a or Wnt11 activated ERK and p38 (fibrotic-related signaling) pathway, promoted the phosphorylation of EGFR, and increased the expression of Frizzled 5 (FZD5) in CFs. Inhibition or knockdown of EGFR greatly attenuated the increased FZD5, p-p38, and p-ERK levels, and the pro-fibrotic effect induced by Wnt5a or Wnt11 in CFs. Si-FZD5 transfection suppressed the increased p-EGFR level, and the fibrotic-related effects in CFs treated with Wnt5a or Wnt11. In conclusion, pressure overload enhances the secretion of Wnt5a or Wnt11 from CMs and CFs which promotes cardiac fibrosis by activation the crosstalk of FZD5 and EGFR. Thus, Wnt5a or Wnt11 may be a novel therapeutic target for the prevention of cardiac fibrosis under pressure overload.Subject terms: Heart failure, Translational research     

UPA, a universal protein array system for quantitative detection of protein–protein, protein–DNA, protein–RNA and protein–ligand interactions

Protein–protein interactions have been widely used to study gene expression pathways and may be considered as a new approach to drug discovery. Here I report the development of a universal protein array (UPA) system that provides a sensitive, quantitative, multi-purpose, effective and easy technology to determine not only specific protein–protein interactions, but also specific interactions of proteins with DNA, RNA, ligands and other small chemicals. (i) Since purified proteins are used, the results can be easily interpreted. (ii) UPA can be used multiple times for different targets, making it economically affordable for most laboratories, hospitals and biotechnology companies. (iii) Unlike DNA chips or DNA microarrays, no additional instrumentation is required. (iv) Since the UPA uses active proteins (without denaturation and renaturation), it is more sensitive compared with most existing methods. (v) Because the UPA can analyze hundreds (even thousands on a protein microarray) of proteins in a single experiment, it is a very effective method to screen proteins as drug targets in cancer and other human diseases.     

二十碳五烯酸的菌种选育及发酵条件

利用含亚麻子油的斜面培养基连续传代和逐渐降低培养温度,诱导筛选的方法,从大量丝状真菌中选育到一株产二十碳五烯酸（all－cis－5、8、11、14、17－eicosopnthenoicacid）较高的被孢霉菌（Motierellasp．）SM481。研究得到最适培养基及最适培养条件。在最适培养及产二十碳五烯酸条件下,细胞干重和二十碳五烯酸产量分别为28．8g／L和0.127g／L。     

基因组数据库简介

本文以北京大学生物信息中心安装的3个国际著名基因组数据库GDB、GenoList和Ensembl为基础,介绍目前常用的基因组数据库,包括这些数据数据库的内容、数据格式、使用方法,以及用于构建上述数据库的数据库管理系统。 Abstract:A brief introduction to the genome databases GDB,GenoList and Ensembl is given.These databases,mirrored and maintained at the Centre of Bioinformatics,Peking University,provide useful information for genome research.     

表达流感病毒神经氨酸酶基因的重组腺病毒的构建

通过RT-PCR方法扩增流感病毒神经氨酸酶基因,将其克隆到腺病毒穿梭载体pTrackCMV,此重组质粒与腺病毒DNA共转化大肠杆菌BJ5183,通过细菌内同源重组获得重组腺病毒DNA,将其转染293细胞获得重组腺病毒.经PCR证实目的基因已整合至腺病毒基因组中,western blot检测到神经氨酸酶的表达.重组病毒经滴鼻和灌胃两种途径免疫小鼠,结果表明2次免疫后滴鼻组和灌胃组均产生明显的免疫应答反应,滴鼻组的免疫效果优于灌胃组.     

蚓激酶研究进展

蚓激酶是一种新型的溶栓制剂,具有良好的溶血栓治疗效果。综述了至今为止分离到的蚓激酶成分、生物学活性、药物学作用、抑制剂和空间结构分析等方面的研究进展 。     

氧化甾醇结合蛋白相关蛋白家族的研究进展

膜脂是细胞膜的主要组分, 也是参与信号转导的重要信号分子。不同脂质分子在细胞膜上的不均等分布需要特殊类型的通道蛋白和运输蛋白来实现。氧化甾醇结合蛋白相关蛋白(ORPs)是一类非常保守的蛋白分子, 能够对磷脂酰肌醇和固醇等脂类分子进行识别并转运, 参与细胞中的许多生理过程, 包括信号转导、囊泡运输、脂类代谢和非囊泡运输等...     

Improving the Genome Annotation of Rhizoctonia solani Using Proteogenomics

Background Rhizoctonia solani is a pathogenic fungus that causes serious diseases in many crops, including rice, wheat, and soybeans. In crop production, it is very important to understand the pathogenicity of this fungus, which is still elusive. It might be helpful to comprehensively understand its genomic information using different genome annotation strategies.MethodsAiming to improve the genome annotation of R. solani, we performed a proteogenomic study based on the existing data. Based on our study, a total of 1060 newly identified genes, 36 revised genes, 139 single amino acid variants (SAAVs), 8 alternative splicing genes, and diverse post-translational modifications (PTMs) events were identified in R. solani AG3. Further functional annotation on these 1060 newly identified genes was performed through homology analysis with its 5 closest relative fungi.ResultsBased on this, 2 novel candidate pathogenic genes, which might be associated with pathogen-host interaction, were discovered. In addition, in order to increase the reliability and novelty of the newly identified genes in R. solani AG3, 1060 newly identified genes were compared with the newly published available R. solani genome sequences of AG1, AG2, AG4, AG5, AG6, and AG8. There are 490 homologous sequences. We combined the proteogenomic results with the genome alignment results and finally identified 570 novel genes in R. solani.ConclusionThese findings extended R. solani genome annotation and provided a wealth of resources for research on R. solani.     

工业蛋白质理性设计与应用

作为合成生物学与绿色生物制造等领域的底层核心技术,蛋白理性设计可有效解决天然功能元件性能不足等共性挑战,创制高性能人工酶元件。值此天津工业生物研究所(Tianjin Institute of Industrial Biotechnology, TIB)创立10周年之际,文中回顾了研究所在工业蛋白理性设计领域的系列重要工作进展。从酶设计方法学研究、新酶反应设计到生物催化应用等方面进行了分析讨论,并展望了本领域未来发展方向。望借此搭建学术界和产业界与酶理性设计的桥梁,促进新技术、新策略的开发应用,加速融合人工酶的基础研究与产业应用,推动我国生物制造领域的科技创新升级。     

微生物来源的谷氨酰胺转氨酶(mTG)介导的单一定点修饰的PEG IFN-α2a(英文)

PEG修饰被认为是改善重组蛋白药物特性的最有效手段,包括增加蛋白质药物在体内的血浆半衰期,降低免疫原性和抗原性。目前典型的PEG修饰手段为将PEG连接至蛋白质的游离氨基,包括赖氨酸和N-末端,但这种连接缺乏选择性,产物为混合物,活性及工艺稳定性差,难以控制。酶法PEG化修饰能有效克服上述缺点,其中谷氨酰胺转氨酶(TGase)可以作为PEG化定点修饰用酶。文中选择重组人干扰素α2a(IFNα2a)进行酶法修饰反应,通过计算机模拟预测IFNα2a可以在第101位Gln特异性定点修饰。将IFNα2a与40 kDa的Y型PEG在微生物来源的谷氨酰胺转氨酶(mTG)催化下进行定点PEG化修饰。结果显示,mTG可以介导IFNα2a特异性位点Gln的单一定点PEG修饰,产生分子量为58 495.6 Da的PEG-Gln101-IFNα2a分子。圆二色谱结果显示,PEG-Gln101-IFNα2a与未修饰的IFNα2a具有相同的二级结构。SD大鼠药代结果显示,与IFNα2a相比,PEG-Gln101-IFNα2a能有效提高药代动力学参数,强于已上市PEGIFNα2a-PEGASYS?。