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| 1 | 组织透明化技术的研究与应用显示文摘背景:对生物体进行全身细胞分析是生物医学领域的主要挑战之一。组织透明技术结合光学成像和图像处理技术,能够将整个器官或全身快速透明并进行结构和细胞分析,为在生命科学中应用先进的光学技术提供了一个非常有前景的解决方案。目的:分析组织透明技术原理及过程,总结组织透明技术研究进展,介绍组织透明成像技术,讨论组织透明技术在生物医学研究领域中的应用。方法:第一作者以“tissue clearing technique,tissue optical clearing,whole-body imaging,3D imaging” | 田婷 杨朝阳 李晓光 | 2020 | 中国组织工程研究2020，24，21： | 4 |
| 2 | 整体器官的光透明成像方法综述显示文摘现代光学成像技术与荧光标记技术不断发展,为高分辨地获取生物组织三维结构信息提供了重要的工具。然而,大多数生物组织具有不透明特性,限制了光在组织中的穿透深度,进而限制了光学成像技术在大组织或整体器官成像中的应用。近年兴起的组织光透明技术通过多种物理、化学手段降低组织对光的衰减,增加光穿透深度,从而提高光学成像的成像深度与成像质量,为整体组织器官的三维成像提供了全新的思路。本文从离体组织光透明方法、大组织器官标记方法、三维整体成像技术三个方面,对整体器官的光透明成像方法进行综述。 | 俞婷婷 朱 | 2020 | 中国激光2020，47，2： | 3 |
| 3 | Targeted Ptpn11 deletion in mice reveals the essential role of SHP2 in osteoblast differentiation and skeletal homeostasis显示文摘The maturation and function of osteoblasts(OBs)rely heavily on the reversible phosphorylation of signaling proteins.To date,most of the work in OBs has focused on phosphorylation by tyrosyl kinases,but little has been revealed about dephosphorylation by protein tyrosine phosphatases(PTPases).SHP2(encoded by PTPN11)is a ubiquitously expressed PTPase.PTPN11 mutations are associated with both bone and cartilage manifestations in patients with Noonan syndrome(NS)and metachondromatosis(MC),although the underlying mechanisms remain elusive.Here,we report that SHP2 deletion in bone gamma-carboxyglutamate protein-expressing(Bglap+)bone cells leads to massive osteopenia in both trabecular and cortical bones due to the failure of bone cell maturation and enhanced osteoclast activity,and its deletion in Bglap+chondrocytes results in the onset of enchondroma and osteochondroma in aged mice with increased tubular bone length.Mechanistically,SHP2 was found to be required for osteoblastic differentiation by promoting RUNX2/OSTERIX signaling and for the suppression of osteoclastogenesis by inhibiting STAT3-mediated RANKL production by osteoblasts and osteocytes.These findings are likely to explain the compromised skeletal system in NS and MC patients and to inform the development of novel therapeutics to combat skeletal disorders. | Lijun Wang Huiliang Yang Jiahui Huang Shaopeng Pei Liyun Wang Jian Q.Feng Dian Jing Hu Zhao Henry M.Kronenberg Douglas C.Moore Wentian Yang | 2021 | Bone Research2021，9，1： | 3 |
| 4 | IntegrinαEβ7^(+)T cells direct intestinal stem cell fate decisions via adhesion signaling显示文摘Intestinal stem cell(ISC)differentiation is regulated precisely by a niche in the crypt,where lymphocytes may interact with stem and transient amplifying(TA)cells.However,whether and how lymphocyte–stem/TA cell contact affects ISC differentiation is largely unknown.Here,we uncover a novel role of T cell–stem/TA cell contact in ISC fate decisions.We show that intestinal lymphocyte depletion results in skewed ISC differentiation in mice,which can be rescued by T cell transfer.Mechanistically,integrinαEβ7 expressed on T cells binds to E-cadherin on ISCs and TA cells,triggering E-cadherin endocytosis and the consequent Wnt and Notch signaling alterations.BlockingαEβ7−E-cadherin adhesion suppresses Wnt signaling and promotes Notch signaling in ISCs and TA cells,leading to defective ISC differentiation.Thus,αEβ7^(+)T cells regulate ISC differentiation at single-cell level through cell–cell contact-mediatedαEβ7−E-cadherin adhesion signaling,highlighting a critical role of the T cell–stem/TA cell contact in maintaining intestinal homeostasis. | Shiyang Chen Yajuan Zheng Xiaojuan Ran Hui Du Hua Feng Lei Yang Yating Wen Changdong Lin Shihui Wang Mengwen Huang Zhanjun Yan Dianqing Wu Hongyan Wang Gaoxiang Ge An Zeng Yi Arial Zeng Jianfeng Chen | 2021 | Cell Research2021，31，12： | 3 |
| 5 | Insights into skeletal stem cells显示文摘The tissue-resident skeletal stem cells(SSCs),which are self-renewal and multipotent,continuously provide cells(including chondrocytes,bone cells,marrow adipocytes,and stromal cells)for the development and homeostasis of the skeletal system.In recent decade,utilizing fluorescence-activated cell sorting,lineage tracing,and single-cell sequencing,studies have identified various types of SSCs,plotted the lineage commitment trajectory,and partially revealed their properties under physiological and pathological conditions.In this review,we retrospect to SSCs identification and functional studies.We discuss the principles and approaches to identify bona fide SSCs,highlighting pioneering findings that plot the lineage atlas of SSCs.The roles of SSCs and progenitors in long bone,craniofacial tissues,and periosteum are systematically discussed.We further focus on disputes and challenges in SSC research. | Qiwen Li Ruoshi Xu Kexin Lei Quan Yuan | 2022 | Bone Research2022，10，4： | 2 |
| 6 | 亲水性透明化技术在骨组织的应用显示文摘在整体组织上实现细胞及亚细胞水平的三维成像是现代组织病理发展的新趋势。光透明化技术的出现打破了组织表面阻隔光透射的壁垒,结合现代光学成像技术可在组织器官上获取包含细胞及亚细胞层面在内的三维图像信息。亲水性透明化方案具有使用简单、无毒性和对荧光蛋白保护性高等特点,是应用最为广泛的组织透明化工具。但其对硬组织渗透性差,折射率匹配性差,在对包含骨在内的硬组织的应用上受到了极大的限制。本文综述亲水性透明化方案针对硬组织透明的发展历程,以推动透明化技术的革新。 | 吴瀚 李茜 吴科锋 崔燎 叶华 | 2021 | 中华骨质疏松和骨矿盐疾病杂志2021，14，2： | 1 |
| 7 | 3D cellular visualization of intact mouse tooth using optical clearing without decalcification显示文摘Dental pulp is composed of nerves,blood vessels,and various types of cells and surrounded by a thick and hard enamel-dentin matrix.Due to its importance in the maintenance of tooth vitality,there have been intensive efforts to analyze the complex cellularlevel organization of the dental pulp in teeth.Although conventional histologic analysis has provided microscopic images of the dental pulp,3-dimensional (3D) cellular-level visualization of the whole dental pulp in an intact tooth has remained a technically challenging task.This is mainly due to the inevitable disruption and loss of microscopic structural features during the process of mechanical sectioning required for the preparation of the tooth sample for histological observation.To accomplish 3D microscopic observation of thick intact tissue,various optical clearing techniques have been developed mostly for soft tissue,and their application for hard tissues such as bone and teeth has only recently started to be investigated.In this work,we established a simple and rapid optical clearing technique for intact mouse teeth without the time-consuming process of decalcification.We achieved 3D cellular-level visualization of the microvasculature and various immune cell distributions in the whole dental pulp of mouse teeth under normal and pathologic conditions.This technique could be used to enable diverse research methods on tooth development and regeneration by providing 3D visualization of various pulpal cells in intact mouse teeth. | Sujung Hong Jingu Lee Jin Man Kim Sun-Young Kim Hyung-Ryong Kim Pilhan Kim | 2019 | International Journal of Oral Science2019，11，3： | 1 |
| 8 | 用于多尺度高分辨率三维成像的双环光片荧光显微技术显示文摘本文提出了一种无衍射光片荧光显微成像技术,可以方便地对从微米到厘米各种尺寸的多种生物样本进行多尺度三维荧光成像。提出一种双环调控方法以解决传统贝塞尔光片旁瓣过重的问题,该方法能够可调节地产生0.4~5μm之间不同厚度类型的无衍射光片,同时其旁瓣占比被压低到30%以下。基于这种新的调控手段设计搭建了一套多尺度光片荧光显微成像系统。该系统展示出适用性极强的多尺度成像能力,例如:活细胞双色三维动态成像、膨胀细胞三维超分辨成像以及介观尺度下全器官的高通量三维成像。证明该多尺度成像模式能够显著提升光片荧光显微镜的成像效率,由此可以推进细胞生物学、组织病理学、神经科学等多种生物医学相关研究的发展。 | 王鹏 周瑶 赵宇轩 费鹏 | 2022 | 中国光学(中英文)2022，15，6： | 1 |
| 9 | 组织透明化技术在神经退行性疾病中的应用研究进展显示文摘组织透明化技术是一种快速发展的新兴组织学技术,将大块组织、单个器官、啮齿动物全身以及人类大型标本,通过一系列物理化学原理与方法进行处理,在保持其完整性的同时快速实现高度光学透明,为高通量地获取生物组织三维结构信息提供了重要的工具。结合现代光学成像技术和荧光标记技术,可显著提高成像深度和图像对比度,已成为经典组织学技术有前景的替代方案,并广泛应用于多个生命科学领域。本文对主要的脑组织透明化方法学原理及其优缺点、组织透明化技术在神经退行性疾病中的应用等进行综述,从整体水平观察神经元、轴突、髓鞘等精细结构的退行性改变以及异质性病理成分在全脑范围内细胞间的演变过程,为最终转向人类脑组织神经退行性病变研究提供可能。 | 陈小玉 罗连响 潘韵琪 鲍波 | 2022 | 解放军医学杂志2022，47，3： | 1 |
| 10 | 组织透明技术在脑重大疾病研究中的应用进展显示文摘组织透明技术是一种新兴的组织学技术,即将组织块或器官通过一系列物理化学的方法处理后实现高度透明且完整的新兴技术。在结合光学成像、荧光标记等技术下,可对组织或器官内部结构进行进一步观察分析。本文围绕组织透明技术进行综述,重点介绍组织透明技术在脑重大疾病研究方面的应用现况及前景。 | 贺电 李静 赵佳佳 孙海涛 | 2020 | 中华神经医学杂志2020，19，3： | 0 |
| 11 | 双光子显微镜的应用优势与维护要素显示文摘双光子显微镜是结合了激光扫描共聚焦显微镜和双光子激发技术的一种新精密仪器,具有低光毒性、高时空分辨率、高信噪比等优点,广泛应用于脑科学、免疫学、肿瘤、胚胎发育等生物医学相关研究领域。但双光子显微镜操作较复杂,对于新用户而言对设备的选用存在一定困惑。通过阐述双光子显微镜的工作原理、样品前期准备、成像难点及设备使用和日常维护要点,梳理双光子显微镜与其他同类成像类设备在成像原理、配置参数、成像特点及应用领域等方面的区别,总结双光子显微镜的应用特点及优势,为从事医学临床、教学及科研提供参考。 | 李娟 张岚岚 吴珏珩 | 2021 | 中国医学装备2021，18，12： | 0 |
| 12 | 组织透明化技术在硬组织中的应用进展显示文摘组织透明化技术是一项新兴技术,其原理是使用物理或化学的方法,通过匹配折射率使组织达到光学透明。组织透明化技术结合荧光成像及三维重建技术,可在细胞水平对组织的内容物和结构进行三维观察及分析。组织透明化技术多运用于软组织,对硬组织的研究相对较少。近年,很多学者改进了透明化技术,使之适用于骨和牙齿等硬组织。本文将对组织透明化技术在硬组织中的应用进展作一综述。 | 薛菲 赵瑚 袁泉 张士文 | 2021 | 中华口腔医学杂志2021，56，6： | 0 |
| 13 | Use of a tissue clearing technique combined with retrograde trans-synaptic viral tracing to evaluate changes in mouse retinorecipient brain regions following optic nerve crush显示文摘Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are limited by the opacity of brain tissue.In this study,we used an innovative tissue cleaning technique combined with retrograde trans-synaptic viral tracing to observe changes in retinorecipient regions connected to retinal ganglion cells in mice after optic nerve injury.Specifically,we performed light-sheet imaging of whole brain tissue after a clearing process.We found that pseudorabies virus 724(PRV724)mostly infected retinal ganglion cells,and that we could use it to retrogradely trace the retinorecipient regions in whole tissue-cleared brains.Unexpectedly,PRV724-traced neurons were more widely distributed compared with data from previous studies.We found that optic nerve injury could selectively modify projections from retinal ganglion cells in the hypothalamic paraventricular nucleus,intergeniculate leaflet,ventral lateral geniculate nucleus,central amygdala,basolateral amygdala,Edinger-Westphal nucleus,and oculomotor nucleus,but not the superior vestibular nucleus,red nucleus,locus coeruleus,gigantocellular reticular nucleus,or facial nerve nucleus.Our findings demonstrate that the tissue clearing technique,combined with retrograde trans-synaptic viral tracing,can be used to objectively and comprehensively evaluate changes in mouse retinorecipient regions that receive projections from retinal ganglion cells after optic nerve injury.Thus,our approach may be useful for future estimations of optic nerve injury and regeneration. | Zong-Yi Zhan Yi-Ru Huang Lu-Wei Zhao Ya-Dan Quan Zi-Jing Li Di-Fang Sun Ya-Li Wu Hao-Yuan Wu Zi-Tian Liu Kai-Li Wu Yu-Qing Lan Min-Bin Yu | 2023 | Neural Regeneration Research2023，18，4： | 0 |
| 14 | 组织光透明技术在骨透明化中的生物医学成像应用与展望显示文摘近年来组织光透明技术的飞速发展,为现代骨科临床基础研究带来了新的契机。组织光透明技术主要通过多种物理、化学手段,降低组织的光散射和光吸收,使光能在组织中更好地传播,从而增加光学成像的深度和对比度。结合多种荧光标记策略,实现更深层、更高分辨的骨组织及三维空间微结构信息,为突破骨这种高散射组织及骨疾病的分子影像学研究带来新的视角与方法。对组织光透明技术的原理及机制进行介绍,并重点从骨组织光透明成像技术的应用现状、新方法及透明机制三个方面加以详述,最后对该技术应用于骨及骨疾病的分子影像学研究前景进行展望。 | 周康为 杨林海 邱棋 郑翔隆 蒋位财 林李嵩 赵庆亮 | 2021 | 激光与光电子学进展2021，58，20： | 0 |
| 15 | Whole-brain Optical Imaging:A Powerful Tool for Precise Brain Mapping at the Mesoscopic Level显示文摘The mammalian brain is a highly complex network that consists of millions to billions of densely-interconnected neurons.Precise dissection of neural circuits at the mesoscopic level can provide important structural information for understanding the brain.Optical approaches can achieve submicron lateral resolution and achieve“optical sectioning”by a variety of means,which has the natural advantage of allowing the observation of neural circuits at the mesoscopic level.Automated whole-brain optical imaging methods based on tissue clearing or histological sectioning surpass the limitation of optical imaging depth in biological tissues and can provide delicate structural information in a large volume of tissues.Combined with various fluorescent labeling techniques,whole-brain optical imaging methods have shown great potential in the brain-wide quantitative profiling of cells,circuits,and blood vessels.In this review,we summarize the principles and implementations of various whole-brain optical imaging methods and provide some concepts regarding their future development. | Tao Jiang Hui Gong Jing Yuan | 2023 | Neuroscience Bulletin2023，39，12： | 0 |
| 16 | 全脑显微光学成像显示文摘全脑介观神经联接研究是解析脑认知功能的神经输入输出环路结构基础、普查基因表达与细胞类型,以及绘制全景立体脑图谱的科学前沿。光学成像方法在横向方向能够达到亚微米的分辨率,并可通过多种手段实现“光学切片”的效果,具备在介观水平观测神经环路的天然优势。基于组织透明或机械切削的自动化全脑显微光学成像方法,突破了光学成像在生物组织中成像深度的限制,具有在大范围内提供介观水平精细观察的技术优势。结合各类生物样本荧光标记技术,全脑显微光学成像方法在神经环路的结构和功能的研究方面有着巨大潜力,已成为剖析全脑神经及血管网络的最佳方式。为了更全面地了解和认识这种有力的工具,总结了近年来发展的各类全脑显微光学成像方法,并展望了未来的技术发展。 | 江涛 龚辉 骆清铭 袁菁 | 2023 | 中国激光2023，50，3： | 0 |
| 17 | Type Ⅱ collagen-positive progenitors are important stem cells in controlling skeletal development and vascular formation显示文摘Type II collagen-positive(Col2^(+))cells have been reported as skeletal stem cells(SSCs),but the contribution of Col2^(+)progenitors to skeletal development both prenatally and postnatally during aging remains unclear.To address this question,we generated new mouse models with ablation of Col2^(+)cells at either the embryonic or postnatal stages.The embryonic ablation of Col2^(+)progenitors resulted in the death of newborn mice due to a decrease in skeletal blood vessels,loss of all vertebral bones and absence of most other bones except part of the craniofacial bone,the clavicle bone and a small piece of the long bone and ribs,which suggested that intramembranous ossification is involved in long bone development but does not participate in spine development.The postnatal ablation of Col2^(+)cells resulted in mouse growth retardation and a collagenopathy phenotype.Lineage tracing experiments with embryonic or postnatal mice revealed that Col2^(+)progenitors occurred predominantly in the growth plate(GP)and articular cartilage,but a limited number of Col2^(+)cells were detected in the bone marrow.Moreover,the number and differentiation ability of Col2^(+)progenitors in the long bone and knee joints decreased with increasing age.The fate-mapping study further revealed Col2^(+)lineage cells contributed to,in addition to osteoblasts and chondrocytes,CD31^(+)blood vessels in both the calvarial bone and long bone.Specifically,almost all blood vessels in calvarial bone and 25.4%of blood vessels in long bone were Col2^(+)lineage cells.However,during fracture healing,95.5%of CD31^(+)blood vessels in long bone were Col2^(+)lineage cells.In vitro studies further confirmed that Col2^(+)progenitors from calvarial bone and GP could form CD31^(+)vascular lumens.Thus,this study provides the first demonstration that intramembranous ossification is involved in long bone and rib development but not spine development.Col2^(+)progenitors contribute to CD31^(+)skeletal blood vessel formation,but the percentage differs between long bone and skull bone.The number and differentiation ability of Col2^(+)progenitors decreases with increasing age. | Xinhua Li Shuting Yang Gongsheng Yuan Dian Jing Ling Qin Hu Zhao Shuying Yang | 2022 | Bone Research2022，10，4： | 0 |
| 18 | 疏水性组织透明化技术的研究进展显示文摘组织透明化技术可通过降低光散射和光吸收使样本在视觉下达到透明,并结合光学显微镜和荧光标记技术实现组织结构三维重建,弥补传统组织切片技术的不连续性、图像重建困难等缺陷。近年来,组织透明化技术逐渐发展,其中疏水性组织透明化技术因具有操作简单、透明速度快和透明度高等特点而广泛应用,本文从其透明程序、发展和应用三个方面进行综述。 | 李茜 吴科锋 | 2022 | 临床医学研究与实践2022，7，14： | 0 |
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