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Protein & Cell 2012年第7期3卷 521-525页

作者： yanhong xue Wei Zhao Wen Du Xiang Zhang Gang Ji Wang Ying Tao Xu National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of SciencesBeijing 100101China College of Life Science and Technology Huazhong University of Science and TechnologyWuhan 430074China

Insulin granule trafficking is a key step in the secretion of glucose-stimulated insulin from pancreaticβ-*** main feature of type 2 diabetes(T2D)is the failure of pancreaticβ-cells to secrete sufficient amounts of insulin to maintain normal blood glucose *** this work,we developed and applied tomography based on scanning transmission electron microscopy(STEM)to image intact insulin granules in theβ-cells of mouse pancreatic *** three-dimensional(3D)recon-struction,we found decreases in both the number and the grey level of insulin granules in db/db mouse pan-creaticβ-***,insulin granules were closer to the plasma membrane in diabeticβ-cells than in control ***,3D ultra-structural tomography may provide new insights into the pathology of insulin se-cretion in T2D.

关键词： insulin secretory granule diabetes stem tomography 3D reconstruction

Ultra-stable super-resolution fluorescence cryo-microscopy for correlative light and electron cryo-microscopy

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Science China(Life Sciences) 2018年第11期61卷 1312-1319页

作者： Xiaojun Xu yanhong xue Buyun Tian Fengping Feng Lusheng Gu Weixing Li Wei Ji Tao Xu College of Life Science and Technology Huazhong University of Science and TechnologyWuhan 430074China National Laboratoty of Biomacromolecules CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China Center for Biological Instrument Development Core Facility for Protein ResearchInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China College of Life Science University of Chinese Academy of SciencesBeijing 100049 China

Remarkable progress in correlative light and electron cryo-microscopy(cryo-CLEM) has been made in the past decade. A crucial component for cryo-CLEM is a dedicated cryo-fluorescence microscope(cryo-FM). Here, we describe an ultra-stable superresolution cryo-FM that exhibits excellent thermal and mechanical stability. The temperature fluctuations in 10 h are less than0.06 K, and the mechanical drift over 5 h is less than 200 nm in three dimensions. We have demonstrated the super-resolution imaging capability of this system(average single molecule localization accuracy of ～13.0 nm). The results suggest that our system is particularly suitable for long-term observations, such as single molecule localization microscopy(SMLM) and cryogenic super-resolution correlative light and electron microscopy(csCLEM).

关键词： cryogenic cryostat super-resolution Cryo-FM Cryo-PALM csCLEM

Bulk-like endocytosis plays an important role in the recycling of insulin granules in pancreatic beta cells

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Protein & Cell 2012年第8期3卷 618-626页

作者： Du Wen yanhong xue Kuo Liang Tianyi Yuan Jingze Lu Wei Zhao Tao Xu Liangyi Chen National Key Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of SciencesBeijing 100101China Lab of Cell Secretion and Metabolism Institute of Molecular MedicinePeking University and National Center for Nanoscience and TechnologyBeijing100871China Department of General Surgery Xuanwu HospitalCapital Medical UniversityBeijing 100053China Graduate School of the Chinese Academy of Sciences Beijing 100864China

Although bulk endocytosis has been found in a number of neuronal and endocrine cells,the molecular mechanism and physiological function of bulk endocytosis remain *** pancreatic beta cells,we have observed bulk-like endocytosis evoked both by flash photolysis and trains of ***-like endocytosis is a clathrin-independent process that is facilitated by enhanced extracellular Ca^(2+) entry and suppressed by the inhibition of dynamin ***,defects in bulklike endocytosis are accompanied by hyperinsulinemia in primary beta cells dissociated from diabetic KKAy mice,which suggests that bulk-like endocytosis plays an important role in maintaining the exo-endocytosis balance and beta cell secretory capability.

关键词： bulk-like endocytosis clathrin-independent endocytosis dynamin diabetic KKAy mice

Alterations of the Ca2＋ signaling pathway in pancreatic beta-cells isolated from db/db mice

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Protein & Cell 2014年第10期5卷 783-794页

作者： Kuo Liang Wen Du Jingze Lu Fei Li Lu Yang yanhong xue Bertil Hille Liangyi Chen Department of General Surgery XuanWu Hospital Capital Medical University Beijing 100053 China National Key Laboratory of Biomacromolecules Institute of Biophysics Chinese Academy of Science Beijing 100101 China The State Key Laboratory of Biomembrane and Membrane Biotechnology Beijing Key Laboratory of CardiometabolicMolecular Medicine Institute of MolecuLar Medicine Peking University Beijing 100871 China Department of Physiology and Biophysics University of Washington School of Medicine Seattle WA 98195 USA

Upon glucose elevation, pancreatic beta-cells secrete insulin in a Ca2＋-dependent manner. In diabetic animal models, different aspects of the calcium signaling path- way in beta-cells are altered, but there is no consensus regarding their relative contributions to the development of beta-cell dysfunction. In this study, we compared the increase in cytosolic Ca2＊（[Ca2＊]~） via Ca2＋ influx, Ca2＊ mobilization from endoplasmic reticulum （ER） calcium stores, and the removal of Ca2＋ via multiple mechanisms in beta-cells from both diabetic db/db mice and non- diabetic C57BL/6J mice. We refined our previous quan- titative model to describe the slow [Ca2＋]i recovery after depolarization in beta-cells from db/db mice. According to the model, the activity levels of the two subtypes of the sarco-endoplasmic reticulum Ca2＋-ATPase （SERCA） pump, SERCA2 and SERCA3, were severely down-reg- ulated in diabetic cells to 65% and 0% of the levels in normal cells. This down-regulation may lead to a reduc- tion in the Ca2＋ concentration in the ER, a compensatory up-regulation of the plasma membrane Na＋/Ca2＋ exchanger （NCX） and a reduction in depolarizationevoked Ca2＋ influx. As a result, the patterns of glucosestimulated calcium oscillations were significantly different in db/db diabetic beta-cells compared with normal cells. Overall, quantifying the changes in the calcium signaling pathway in db/db diabetic beta-cells will aid in the development of a disease model that could provide insight into the adaptive transformations of beta-cell function during diabetes development.

关键词： diabetic beta-cells calcium signalingalterations SERCA pump db/db mice

Cryogenic superresolution correlative light and electron microscopy of vitreous sections

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Biophysics Reports 2022年第4期8卷 193-204页

作者： Buyun Tian Maoge Zhou Fengping Feng Xiaojun Xu Pei Wang Huiqin Luan Wei Ji yanhong xue Tao Xu National Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences University of Chinese Academy of Sciences National Research Center for Rehabilitation Technical Aids

Fluorescence microscopy and electron microscopy complement each other as the former provides labelling and localisation of specific molecules and target structures while the latter possesses excellent revolving power of fine structure in context. These two techniques can combine as correlative light and electron microscopy(CLEM) to reveal the organisation of materials within the cell. Frozen hydrated sections allow microscopic observations of cellular components in situ in a near-native state and are compatible with superresolution fluorescence microscopy and electron tomography if sufficient hardware and software support is available and a well-designed protocol is followed. The development of superresolution fluorescence microscopy greatly increases the precision of fluorescence annotation of electron tomograms. Here, we provide detailed instructions on how to perform cryogenic superresolution CLEM on vitreous sections. From fluorescence-labelled cells to high pressure freezing, cryoultramicrotomy, cryogenic single-molecule localisation microscopy, cryogenic electron tomography and image registration, electron tomograms with features of interest highlighted by superresolution fluorescence signals are expected to be obtained.

关键词： Electron microscopy Electron tomography Fluorescence microscopy Cryogenic Superresolution Correlative light and electron microscopy (CLEM)

TGF-βchanges cyto/mito-ribosome balance to target respiratory chain complex V biogenesis in pulmonary fibrosis therapy

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Signal Transduction and Targeted Therapy 2023年第4期8卷 1384-1387页

作者： Na Zhang Zunling Zhao Yin Zhao Lei Yang yanhong xue Yun Feng Jianjun Luo Runsheng Chen Wei Wei Yan Qin Department of Rheumatology and Immunology Tianjin Medical University General Hospital300052 TianjinChina CAS Center for Excellence in Biomacromolecules Institute of BiophysicsChinese Academy of Sciences15 Datun RoadChaoyang District100101 BeijingChina University of Chinese Academy of Sciences 100049 BeijingChina

Dear Editor,Pulmonary fibrosis(PF)is characterized by failed alveolar reepithelialization and fibroblast activation.1 A continuing struggle in the field has been how to diagnose and treat the disease early and effectively on the basis of shared pathogenetic *** growth factor-β(TGF-β)signaling and mitochondria are involved in PF pathogenesis.2 Mitochondrial respiratory chain complexes(RCCs)I,III,IV,and V comprise both nuclear DNA(nDNA)-and mitochondrial DNA(mtDNA)-encoded subunits,and their biogenesis depends on the cooperation of cytoplasmic(cyto)and mitochondrial(mito)translation(Supplementary Fig.1).3 To date,a correlation between PF and RCC biogenesis has not been reported.

关键词： respiratory genesis struggle

Spatial distribution of IL4 controls iNKT cell-DC crosstalk in tumors

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Cellular & Molecular Immunology 2020年第5期17卷 496-506页

作者： Lu Wang Zhilan Liu Lili Wang Qielan Wu Xiang Li Di Xie Huimin Zhang Yongdeng Zhang Lusheng Gu yanhong xue Ting Yue Gang Liu Wei Ji Haiming Wei Tao Xu Li Bai Division of Life Sciences and Medicine Department of Oncology of The First Affiliated HospitalThe CAS Key Laboratory of Innate Immunity and Chronic DiseaseUniversity of Science and Technology of China230027 HefeiChina National Key Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of Sciences100101 BeijingChina School of Life Sciences University of Science and Technology of China230027 HefeiChina National Synchrotron Radiation Laboratory University of Science and Technology of China230027 HefeiChina

The spatiotemporal distribution of cytokines orchestrates immune responses in vivo,yet the underlying mechanisms remain to be *** showed here that the spatial distribution of interleukin-4(IL4)in invariant natural killer T(iNKT)cells regulated crosstalk between iNKT cells and dendritic cells(DCs)and controlled iNKT cell-mediated T-helper type 1(Th1)*** persistent polarization of IL4 induced by strong lipid antigens,that is,α-galactosylceramide(αGC),caused IL4 accumulation at the immunological synapse(IS),which promoted the activation of the IL4R-STAT6(signal transducer and activator of transcription 6)pathway and production of IL12 in DCs,which enhanced interferon-γ(IFNγ)production in iNKT ***,the nonpolarized secretion of IL4 induced by Th2 lipid antigens with a short or unsaturated chain was incapable of enhancing this iNKT cell-DC crosstalk and thus shifted the immune response to a Th2-type *** nonpolarized secretion of IL4 in response to Th2 lipid antigens was caused by the degradation of Cdc42 in iNKT ***,reduced Cdc42 expression was observed in tumorinfiltrating iNKT cells,which impaired IL4 polarization and disturbed iNKT cell-DC crosstalk in tumors.

关键词： Spatial distribution Polarization IL4 iNKT DC Crosstalk Tumor

基于89C51+DS18B20的智能温控器的研制

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微计算机信息 2007年第3S期23卷 198-199页

作者：薛燕红陕西理工学院

介绍了新型单总线结构温度传感器DSl8B20的结构特征、工作原理及控制方法,阐述了以89C205l为单片机和以DSl8B20为传感器构成的智能温度控制器的电路组成、工作原理、程序设计,说明了在研制过程中总结出来的使用注意事项。该温控器可广... 详细信息

介绍了新型单总线结构温度传感器DSl8B20的结构特征、工作原理及控制方法,阐述了以89C205l为单片机和以DSl8B20为传感器构成的智能温度控制器的电路组成、工作原理、程序设计,说明了在研制过程中总结出来的使用注意事项。该温控器可广泛应用于人门日常生活、工农业生产和科学研究领域,具有一定的推广价值。

关键词：温度 DS18B20 89C2051 程序控制

磺化聚醚醚酮与壳聚糖共混制备直接甲醇燃料电池用质子交换膜

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高分子学报 2010年第3期20卷 285-291页

作者：薛艳红傅荣强徐铜文中国科学技术大学化学与材料科学学院功能膜实验室合肥230026 新加坡国立大学化学工程系新加坡119260

给出了不同磺化度下的磺化聚醚醚酮(SPEEK)用作质子交换膜的一系列性能,另外提出了一种新型的酸碱共混质子交换膜,其中,磺化聚醚醚酮和壳聚糖分别被选为酸性、碱性高分子电解质,并对所制备的质子交换膜的相关性能如质子传导性,甲醇渗透... 详细信息

给出了不同磺化度下的磺化聚醚醚酮(SPEEK)用作质子交换膜的一系列性能,另外提出了一种新型的酸碱共混质子交换膜,其中,磺化聚醚醚酮和壳聚糖分别被选为酸性、碱性高分子电解质,并对所制备的质子交换膜的相关性能如质子传导性,甲醇渗透性,吸水率以及膜溶胀性、热稳定性等进行了表征,结果表明此种新型复合膜尽管在质子传导性能方面有所下降,阻醇性能改变不大,但是膜溶胀性和吸水率方面有了较大的改善.磺化度为71.4%的SPEEK与壳聚糖以5∶1摩尔比共混制备的质子交换膜,其性质可以与商品化的Nafion 117相媲美,有望在甲醇燃料电池中得到应用.

关键词：质子交换膜直接甲醇燃料电池磺化聚醚醚酮壳聚糖共混膜

A unified deep-learning network to accurately segment insulin granules of different animal models imaged under different electron microscopy methodologies

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Protein & Cell 2019年第4期10卷 306-311页

作者： Xiaoya Zhang Xiaohong Peng Chengsheng Han Wenzhen Zhu Lisi Wei Yulin Zhang Yi Wang Xiuqin Zhang Hao Tang Jianshe Zhang Xiaojun Xu Fengping Feng yanhong xue Erlin Yao Guangming Tan Tao Xu Liangyi Chen State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing ChinaState Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China Drug Discovery Center Key Laboratory of Chemical Genomics Peking University Shenzhen Graduate School Shenzhen China State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China State Key Laboratory of Membrane Biology Beijing Key Laboratory of Cardiometabolic Molecular Medicine Institute of Molecular Medicine Peking University Beijing China University of Chinese Academy of Sciences Beijing China Marine Science College of Zhejiang Ocean University National Engineering Research Center of Marine Facilities Aquaculture Zhoushan China National Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences Beijing China National Laboratory of Biomacromo

Insulin is important for body metabolism regulation and glucose homeostasis,and its dysregulation often leads to metabolic syndrome(MS)and *** is normally stored in large dense-core vesicles(LDCVs)in pancreatic beta cells,and significant reductions in the number,size,gray level and density of insulin granules confer diabetes both in mice(xue et al.,2012)and humans(Masini et al.,2012).

关键词： deep-learning network animal