piR-823 inhibits cell apoptosis via modulating mitophagy by binding to PINK1 in colorectal cancer

Mitophagy plays a vital role in the maintenance of mitochondrial homeostasis and tumorigenesis. Noncoding RNA piR-823 contributes to colorectal tumorigenesis. In this study, we aim to evaluate piR-823-mediated mitophagy and its mechanistic association with colorectal cancer (CRC). Digital gene expression analysis was performed to explore the potential functions of piR-823. A piR-823 antagomir (Ant-823) was used to inhibit piR-823 expression, and piR-823 mimics (mimics-823) were used to increase piR-823 expression. Mitophagy was measured in vivo and in vitro by immunofluorescence and western blot analysis. JC-1 staining, ATP production, real-time PCR, and western blot analysis were used to measure changes in mitochondrial quality and number. siRNA transfection was used to inhibit mitophagy, and CCCP was used to induce mitophagy. RNA pull-down assays and RNA-binding protein immunoprecipitation assays were conducted to investigate the molecular mechanisms. Here, we found that CRC cells transfected with Ant-823 presented an altered expression of autophagic and mitophagy genes by Digital gene expression analysis. Ant-823 could promote Parkin activation and mitophagy in vitro and in vivo, followed by mitochondrial loss and dysfunction of some mitochondria, whereas mimics-823 exerted the opposite effects in CRC cells. The inhibition of mitophagy by siParkin alleviated Ant-823-induced mitochondrial loss and dysfunction, as well as apoptosis to a certain extent. Furthermore, piR-823 was found to interact with PINK1 and promote its ubiquitination and proteasome-dependent degradation, thus alleviating mitophagy. Finally, these findings were verifed in samples obtained by patients affected by colorectal cancer. In conclusion, we identify a novel mechanism by which piR-823 regulates mitophagy during CRC tumorigenesis by increasing PINK1 degradation. Subject terms: Colorectal cancer, Gastrointestinal cancer     

副溶血性弧菌重复序列-PCR分型研究

利用基因外重复回文序列-PCR(REP-PCR)和肠细菌基因间共有重复序列-PCR(ERIC-PCR)技术,对副溶血性弧菌进行了分子分型研究和亲缘关系的探讨,并使用Hunter和Gaston方法计算分辨力指数.结果显示40株副溶血性弧茵分离株均可扩增产生可重复的DNA指纹图谱,并且不同菌株基因组DNA的扩增条带具有多态性.根据SPSS10.0软件得出的树状图结果,REP-PCR可以把40株茵分为21个型,分辨力指数可达到0.953,优势菌型为G1型;ERIC-PCR可将40株菌分成4个型,分辨力指数为0.5.研究显示重复序列-PCR方法可以用于该菌分型分析,REP-PCR具有较好的分型能力.在两种PCR的DNA指纹图谱中,血清型O1群与O3群主条带均非常相似,表明它们之间亲缘关系密切.     

亚热带天然常绿阔叶林林下9种灌木细根形态和C、N化学计量特征

林下灌木是亚热带常绿阔叶林重要的构成部分,但林下灌木细根功能性状变异规律及地下生态策略仍不清楚。以福建建瓯万木林自然保护区内9种灌木为研究对象,对细根直径、根长、比根长、组织密度、碳浓度和氮浓度6个细根性状进行研究,采用序级划分法,分析不同树种细根性状序级间的变化特征、常绿和落叶灌木细根性状之间的差异,不同序级细根性状之间的关系以及细根性状变异维度。结果表明:树种和序级对9种灌木细根形态和化学性质有显著影响。直径、根长、根组织密度随着序级的增加而逐渐增加,比根长和氮浓度逐渐减小,碳浓度在序级间的变化趋势不一,未表现出明显的规律。落叶灌木细根直径、根长和氮浓度均显著高于常绿灌木,碳浓度和组织密度显著低于常绿灌木,表明与常绿灌木相比落叶灌木更偏向于资源获取型生态策略,常绿灌木则更偏向于保守型策略。灌木细根在不同序级间的直径与比根长、组织密度,氮浓度与组织密度有较强的相关性,细根其他性状间的关系并不密切或因序级而异。主成分分析结果表明灌木细根性状变异沿一个主成分轴发生变异,该轴表示灌木细根的资源获取和保守的权衡策略。     

乳酸菌微机检索软件的开发

以现有乳酸菌信息为依据,利用汉字foxbase^＋2．10进行数据处理,研制出一套乳酸菌微机检索软件。该软件的特点：检索速度快,途径灵活,增添、修改、删除、打印方便,各模块性能优良,实用性强。     

Effect of a Functional Phospholipid Metabolome-Protein Association Pathway on the Mechanism of COVID-19 Disease Progression

This study aimed to explore the clinical practice of phospholipid metabolic pathways in COVID-19. In this study, 48 COVID-19 patients and 17 healthy controls were included. Patients were divided into mild (n=40) and severe (n=8) according to their severity. Phospholipid metabolites, TCA circulating metabolites, eicosanoid metabolites, and closely associated enzymes and transfer proteins were detected in the plasma of all individuals using metabolomics and proteomics assays, respectively. 30 of the 33 metabolites found differed significantly (P<0.05) between patients and healthy controls (P<0.05), with D-dimmer significantly correlated with all of the lysophospholipid metabolites (LysoPE, LysoPC, LysoPI and LPA). In particular, we found that phosphatidylinositol (PI) and phosphatidylcholine (PC) could identify patients from healthy controls (AUC 0.771 and 0.745, respectively) and that the severity of the patients could be determined (AUC 0.663 and 0.809, respectively). The last measurement before discharge also revealed significant changes in both PI and PC. For the first time, our study explores the significance of the phospholipid metabolic system in COVID-19 patients. Based on molecular pathway mechanisms, three important phospholipid pathways related to Ceramide-Malate acid (Cer-SM), Lysophospholipid (LPs), and membrane function were established. Clinical values discovered included the role of Cer in maintaining the inflammatory internal environment, the modulation of procoagulant LPA by upstream fibrinolytic metabolites, and the role of PI and PC in predicting disease aggravation.     

生态环境移动数据采集系统研究与实现

针对快速、实时、有效采集并录入生态环境野外调查大样本量、多源数据的需求,充分应用移动GIS技术、移动智能终端、3G等现代信息技术优势,提出了基于ArcGIS for Mobile的移动数据采集方案,研究解决了包括系统运行机制、数据访问模式、移动数据库技术等面向全国生态环境野外调查的移动GIS关键技术,设计并研发野外调查移动数据采集系统。该系统采用C#和Java语言,以SQL Server 2008 为服务器端的数据库环境,在Microsoft Visual Studio 2008集成开发环境上实现设计开发,并在全国生态环境10年变化遥感调查与评估项目土地覆盖类型地面核查、生态系统参数野外观测等工作中予以应用。实践检验表明：该系统实现了野外调查数据的数字采集、智能校验、实时上传与有效管理,简化了填报程序,规范了填报内容,提高了工作效率,能够为生态环境相关的调查数据采集提供信息化支持。     

体外重构STUB1介导α-1抗胰蛋白酶突变体Z蛋白泛素化修饰反应体系

α-1抗胰蛋白酶Z型突变体蛋白(α-1 antitrypsin Z-mutant protein, ATZ)是引发α-1抗胰蛋白酶缺陷症(α-1 antitrypsin deficiency, AATD)的主要原因,研究ATZ蛋白的泛素化修饰和降解对于治疗AATD具有重要意义。STUB1是一种重要的E3泛素连接酶,参与调节多种蛋白质的泛素化修饰。然而,STUB1是否参与ATZ的泛素化修饰尚未明确。本研究首先将ATZ和STUB1的编码基因克隆到pET28a质粒,构建了这2个蛋白的表达质粒。随后,将重组质粒转入大肠杆菌表达系统,在优化诱导条件实现了重组蛋白的异源表达。通过金属螯合亲和层析技术纯化得到目的蛋白,并通过蛋白质谱分析验证了其氨基酸序列的准确性。利用纯化的ATZ和STUB1重组蛋白,构建了一个体外泛素化修饰反应体系。实验结果显示,在ATP、E1泛素激活酶和E2泛素结合酶的协同作用下,STUB1成功催化了ATZ的泛素化修饰。本研究提供了一种体外获得Z型突变体ATZ纯化蛋白的方法,并确认了STUB1介导ATZ的泛素化修饰功能,推进了对α-1抗胰蛋白酶Z型突变体蛋白在细胞内降解过程的调控机制的理解。     

Quantitative regulation of the thermal stability of enveloped virus vaccines by surface charge engineering to prevent the self-aggregation of attachment glycoproteins

The development of thermostable vaccines can relieve the bottleneck of existing vaccines caused by thermal instability and subsequent poor efficacy, which is one of the predominant reasons for the millions of deaths caused by vaccine-preventable diseases. Research into the mechanism of viral thermostability may provide strategies for developing thermostable vaccines. Using Newcastle disease virus (NDV) as model, we identified the negative surface charge of attachment glycoprotein as a novel determinant of viral thermostability. It prevented the temperature-induced aggregation of glycoprotein and subsequent detachment from virion surface. Then structural stability of virion surface was improved and virus could bind to and infect cells efficiently after heat-treatment. Employing the approach of surface charge engineering, thermal stability of NDV and influenza A virus (IAV) vaccines was successfully improved. The increase in the level of vaccine thermal stability was determined by the value-added in the negative surface charge of the attachment glycoprotein. The engineered live and inactivated vaccines could be used efficiently after storage at 37°C for at least 10 and 60 days, respectively. Thus, our results revealed a novel surface-charge-mediated link between HN protein and NDV thermostability, which could be used to design thermal stable NDV and IAV vaccines rationally.