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A quantitative method to assess bacterial adhesion using recombinant bioluminescent Pseudomonas aeruginosa

Biophysics Reports 2021年第1期7卷 55-70页

作者： Lu Wang xinhua qiao Lei Gao Chang Chen Yi Wan Microbiology Institute of Shaanxi National Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences University of Chinese Academy of Sciences Beijing Institute for Brain Disorders

Bioluminescence technology has been widely used in the field of medical detection. The bioluminescent lux reporter system provides a non-invasive platform to monitor bacterial growth and expression in real time. This study aimed to establish a method for detecting bacterial adhesion on the surface of materials, including medical devices, by using recombinant bioluminescent Pseudomonas aeruginosa containing a lux reporter. By monitoring the growth and bioluminescent properties of the recombinant PAO1-lux strain, the optimal test conditions for bacterial adhesion detection in vitro were determined to be as follows: an initial inoculation density of 10~5 to 10~6 CFU/m L, M9 medium at a p H 6.2, an adhesion time of 6 h, and the collection of adherent bacteria by ultrasonic cleaning. The traditional CFU counting method and the bioluminescence method were compared, and the applicability of the new method was verified by testing the adhesion of bacteria on the surface of various materials. The validated bioluminescent strains could serve as strong candidates to be used as bacterial detection tools in applications such as bacterial adhesion evaluation as well as supplements and alternatives to traditional microbiological testing procedures. In addition, this method has the potential to enable the study of bacterial adhesion on the surface of inanimate objects and living tissues. With the development of this method and its wide applicability, it is expected to become a standard method for the detection of bacterial adhesion and the screening of anti-adhesion materials.

关键词： Bacterial adhesion PAO1-lux Bioluminescence assay Medical materials

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Redox-stress response resistance(RRR)mediated by hyperoxidation of peroxiredoxin 2 in senescent cells

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Science China(Life Sciences) 2023年第10期66卷 2280-2294页

作者： Jiao Meng Yuanyuan Wang Zhenyu Lv xinhua qiao Aojun Ye qiaoli Zhu Chang Chen National Laboratory of Biomacromolecules CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Beijing Institute for Brain Disorders Capital Medical UniversityBeijing 100069China

Aging is closely related to redox *** our previous work,we proposed a new concept,“redox-stress response capacity(RRC),”and found that the decline in RRC was a dynamic characteristic of ***,the mechanism of RRC decline during aging remains *** this study,using the senescent human fibroblast cell model and Caenorhabditis elegans model,we identified that peroxiredoxin 2(PRDX2),as a hydrogen peroxide(H_(2)O_(2))sensor,was involved in mediating ***2 knockdown led to a decline of RRC and accelerated senescence in fibroblasts and prdx-2 mutant *** also showed decreased *** mechanism study showed that the decreased sensor activity of PRDX2 was related to the increase in hyperoxidation of PRDX2 in senescent ***,the level of PRDX2 hyperoxidation also increased in old *** overexpression of both PRDX2 and sulfiredoxin(SRX)rescued the reduced RRC and delayed *** increase in PRDX2 hyperoxidation in senescent cells led to a decrease in its sensor activity,resulting in the decreased cellular response to H_(2)O_(2),which is similar to the mechanism of insulin resistance due to the lower insulin receptor *** of young cells with a high level of H_(2)O_(2)to induce a higher level of PRDX2-SO_(3) resulted in mimicking the RRC decline in senescent cells,which is also similar to a model of insulin resistance induced by high levels of *** these results thrillingly indicate that there is an insulin-resistance-like phenomenon in senescent cells,we named it redox-stress response resistance,*** in senescent cells is an important new discovery that explains RRC decline during aging and reveals the internal relationship between redox regulation and aging from a new perspective.

关键词： peroxiredoxin 2(PRDX2) redox-stress response capacity(RRC) redox-stress response resistance(RRR) H_(2)O_(2)sensor redox relay aging oxidative stress

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SNObase,a database for S-nitrosation modification

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Protein & Cell 2012年第12期3卷 929-933页

作者： Xu Zhang Bo Huang Lunfeng Zhang Yuying Zhang Yingying Zhao Xiaofei Guo xinhua qiao Chang Chen National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of SciencesBeijing 100101China Graduate School of the Chinese Academy of Sciences Beijing 100049China Department of Physiology and Pathophysiology Key Laboratory of Molecular Cardiovascular Sciences of Education MinistryPeking University Health Science CenterBeijing 100083China

S-Nitros(yl)ation is a ubiquitous redox-based post-translational modification of protein cysteine thiols by nitric oxide or its derivatives,which transduces the bioactivity of nitric oxide(NO)by regulation of protein conformation,activity,stability,localization and pro-tein-protein *** years,more and more S-nitrosated proteins were identified in physiological and pathological processes and the number is still *** we developed a database named SNO-base(http://***),which collected S-nitrosation targets extracted from literatures up to June 1st,*** contained 2561 instances,and provided information about S-nitrosation targets,sites,biological model,related diseases,trends of S-nitrosation level and effects of S-nitrosation on protein *** SNObase,we did functional analysis for all the SNO targets:In the gene ontology(GO)biological process category,some processes were discovered to be related to S-nitrosation(“response to drug”,“regulation of cell motion”)besides the previously reported related *** the GO cellular component category,cytosol and mitochondrion were both *** the KEGG pathway enrichment results,we found SNO targets were enriched in different diseases,which suggests possible significant roles of S-nitrosation in the progress of these *** SNObase means to be a database with precise,comprehensive and easily accessible information,an environment to help researchers integrate data with comparison and relevancy analysis between different groups or works,and also an SNO knowledgebase offering feasibility for systemic and global analysis of S-nitrosation in interdisciplinary studies.

关键词： S-nitrosation S-nitrosylation database S-Nitrosothiol(SNO)

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PI4KIIα regulates insulin secretion and glucose homeostasis via a PKD-dependent pathway

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Biophysics Reports 2018年第1期4卷 25-38页

作者： Lunfeng Zhang Jiangmei Li Panpan Zhang Zhen Gao Yingying Zhao xinhua qiao Chang Chen National Laboratory of Biomacromolecules CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of Sciences University of Chinese Academy of Sciences Shanghai Institute for Advanced Immunochemical Studies Shanghai Tech University Beijing Institute for Brain Disorders

Insulin release by pancreatic b cells plays a key role in regulating blood glucose levels in humans, and to understand the mechanism for insulin secretion may reveal therapeutic strategies for diabetes. We found that PI4KIIα transgenic(TG) mice have abnormal glucose tolerance and higher serum glucose levels than wild-type mice. Glucose-stimulated insulin secretion was significantly reduced in both PI4KIIα TG mice and PI4KIIα-overexpressing pancreatic b cell lines. A proximity-based biotin labeling technique, Bio ID, was used to identify proteins that interact with PI4KIIα, and the results revealed that PI4KIIα interacts with PKD and negatively regulates its activity. The effect of PI4KIIα on insulin secretion was completely rescued by altering PKD activity. PI4KIIα overexpression also worsened glucose tolerance in streptozotocin/high-fat diet-induced diabetic mice by impairing insulin *** study has shed new light on PI4KIIα function and mechanism in diabetes and identified PI4KIIα as an important regulator of insulin secretion.

关键词： Carriers of the trans-Golgi network to the cell surface (CARTS) Insulin secretion Phosphatidylinositol 4-kinase IIα (PI4KIIα) Protein kinase D (PKD)

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Reductive stress induced by Ero1α S-nitrosation in endoplasmic reticulum accelerates cell senescence

Reductive stress induced by Ero1α S-nitrosation in endoplas...

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中国生物化学与分子生物学会2019年全国学术会议暨学会成立四十周年

作者： xinhua qiao Chang Chen National Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences University of Chinese Academy of Sciences

Redox homeostasis in cells is crucial for the function of biomacromolecules. Oxidative stress is known to mediate many cellular signal transduction in physiological and pathological processes, however, the biological effects and mechanisms of reductive stress(RS, an abnormal increase in electron pressure or reducing equivalents(GSH/GSSG;NADH/NAD;NADPH/NADP) are still poorly understood. In this study, we found that the ER exhibited reductive stress in senescent cells, and reductive stress accelerated senescence. Further study showed that the decrease of Ero1α activity is an important cause of ER reductive stress in the process of aging, and delaying aging was successfully achieved by increasing the oxidative power in the ER. For mechanism study, we found the Ero1α S-nitrosation increased in senescence cells. Ero1α C166 and C131 were identified as the site of S-nitrosation and result in decreasing of Ero1α activity. Moreover, we found that ER reductive stress led to protein homeostatic imbalance and decreased ER folding ability which accelerated aging. Our results suggested a new mechanism of aging caused by insufficient oxidation in the ER.

关键词： Reductive Stress ER Ero1α S-nitrosation Cell senescence

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Exploring the precision redox map during fasting-refeeding and satiation in ***

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Stress Biology 2023年第1期3卷 187-200页

作者： xinhua qiao Lu Kang Chang Shi Aojun Ye Dongli Wu Yuyunfei Huang Minghao Deng Jiarui Wang Yuzheng Zhao Chang Chen National Laboratory of Biomacromolecules CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China School of Basic Medical Sciences of Southwest Medical University Luzhou 646000China University of Chinese Academy of Sciences Beijing 100049China School of Pharmacy East China University of Science and TechnologyShanghai 200237China

Fasting is a popular dietary strategy because it grants numerous advantages,and redox regulation is one mecha-nism ***,the precise redox changes with respect to the redox species,organelles and tissues remain unclear,which hinders the understanding of the metabolic mechanism,and exploring the precision redox map under various dietary statuses is of great *** redox-sensitive *** strains stably expressing genetically encoded redox fluorescent probes(Hyperion sensing H_(2)O_(2) and Grx1-roGFP2 sensing GSH/GSSG)in three organelles(cytoplasm,mitochondria and endoplasmic reticulum(ER))were constructed in two tissues(body wall muscle and neurons)and were confirmed to respond to redox *** H_(2)O_(2) and GSSG/GSH redox changes in two tissues and three organelles were obtained by confocal microscopy during fasting,refeeding,and *** found that under fasting condition,H_(2)O_(2) decreased in most compartments,except for an increase in mitochondria,while GSSG/GSH increased in the cytoplasm of body muscle and the ER of *** refeeding,the redox changes in H_(2)O_(2) and GSSG/GSH caused by fasting were reversed in most organelles of the body wall muscle and *** the sati-ated state,H_(2)O_(2) increased markedly in the cytoplasm,mitochondria and ER of muscle and the ER of neurons,while GSSG/GSH exhibited no change in most organelles of the two tissues except for an increase in the ER of *** study systematically and precisely presents the redox characteristics under different dietary states in living animals and provides a basis for further investigating the redox mechanism in metabolism and optimizing dietary guidance.

关键词： Redox Fasting Refeeding Satiation Hyperion Grx1-roGFP2 Caenorhabditis elegans

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“天下第一廉吏”:清前期一个文化符号的形塑与意涵

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清华大学学报（哲学社会科学版） 2019年第1期34卷 32-43,195页

作者：乔新华山西大学历史文化学院

山西士人于成龙生前是深受康熙褒赞的模范官员,死后留下了"天下第一廉吏"的美名。今人多视其为清官,忽略了"廉吏"也是清初特殊政治环境下的产物。作为汉族士人,他因社会现实和家乡之情的考量而仕清,这是一个充满"政治治理"性格的决定。... 详细信息

山西士人于成龙生前是深受康熙褒赞的模范官员,死后留下了"天下第一廉吏"的美名。今人多视其为清官,忽略了"廉吏"也是清初特殊政治环境下的产物。作为汉族士人,他因社会现实和家乡之情的考量而仕清,这是一个充满"政治治理"性格的决定。其后"忠孝难以两全"困境化解的过程,体现了皇权与士人的密切互动。谕祭和祠祀使他的生平成为建构社会礼制的力量。个人选择、政治权力、社会力量,将"廉吏"塑造成为一个政治化的文化符号。

关键词：于成龙清代前期天下第一廉吏文化符号

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四种鲜为人知的专志与明清以来的洪洞问题研究

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中国地方志 2006年第8期 56-62页

作者：乔新华山西大学历史文化学院 030006

山西洪洞大槐树是无数人心目中故乡的象征,相关研究成果层出不穷。在传统的研究思路下,洪洞大槐树移民作为明初历史上的大事件确立了在中国移民史上的重要地位。作者利用志书等文献史料对传统研究思路提出疑问,从洪洞内部探讨其成为故... 详细信息

山西洪洞大槐树是无数人心目中故乡的象征,相关研究成果层出不穷。在传统的研究思路下,洪洞大槐树移民作为明初历史上的大事件确立了在中国移民史上的重要地位。作者利用志书等文献史料对传统研究思路提出疑问,从洪洞内部探讨其成为故乡象征的深层内涵,认为大槐树移民传说是晚清到民国一个重塑汉族正统性的民间做法,是士绅阶层彰显洪洞历史的一次成功实践。本文在社会史研究的视角下,对洪洞问题给予一个全新的解读。

关键词：志书洪洞研究

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明代五台山寺院经济中的制约因素

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宗教学研究 2020年第4期 103-109页

作者：王国棉乔新华山西大学历史文化学院山西省社会科学院(省政府发展研究中心)

明代五台山寺院经济发展相对处于低潮,寺院群内等级差别明显。这种状况可以从五台山在政治管理序列中的自治地位,以及明代佛教"分寺清宗"运动的政策倾向性中得到说明。明代对佛教规模化、专业化、市场化的改造,直接导致佛教世俗化、社... 详细信息

明代五台山寺院经济发展相对处于低潮,寺院群内等级差别明显。这种状况可以从五台山在政治管理序列中的自治地位,以及明代佛教"分寺清宗"运动的政策倾向性中得到说明。明代对佛教规模化、专业化、市场化的改造,直接导致佛教世俗化、社会化的加强。寺院在山场管理上的自主权,与人口流动带来的劳动力,使得五台山寺院经济虽不如往昔鼎盛,但依然稳步发展。此外,寺产的差异性与不平衡性问题也影响着五台山寺院集体经济活动的发展态势。

关键词：五台山寺院经济世俗化

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以HIV-1 Vif与人APOBEC3G为靶点的抗HIV-1药物的研究方法

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中国中药杂志 2011年第6期36卷 806-809页

作者：乔新华张文俊李泽琳曾毅北京工业大学生命学院北京100124

人载脂蛋白B mRNA编辑酶催化多肽样蛋白3G(apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3protein G,APOBEC3G)是宿主的抗HIV-1(human immunodeficiency virus type 1)因子,而HIV-1辅助蛋白———病毒感染因子Vif(viral infectivity factor)可通过介导蛋白酶体途径降解APOBEC3G,因此针对APOBEC3G及HIV-1Vif进行抑制剂设计已经成为抗HIV-1药物研究新的方向之一,相应用于研究Vif-APOBEC3G相互作用的方法也越来越多,如免疫印迹、免疫杂交、脉冲追踪试验、生物发光共振能量转移检测、BIAcore检测等。作者将目前用于以Vif-APOBEC3G为靶点的药物的筛选及作用机制的研究方法进行了综述,为基于此的研究提供了策略。

关键词： APOBEC3G HIV-1Vif 抗HIV-1抑制剂

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