The structure of intestinal tissue is complex.In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases.Recently,organoids have been successfully constructe...The structure of intestinal tissue is complex.In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases.Recently,organoids have been successfully constructed and they have come to play an important role in biomedical research.Organoids are miniaturized three-dimensional(3D)organs,derived from stem cells,which mimic the structure,cell types,and physiological functions of an organ,making them robust models for biomedical research.Intestinal organoids are 3D micro-organs derived from intestinal stem cells or pluripotent stem cells that can successfully simulate the complex structure and function of the intestine,thereby providing a valuable platform for intestinal development and disease research.In this article,we review the latest progress in the construction and application of intestinal organoids.展开更多
Objective： Although lithium has been a commonly prescribed neurotrophic/neuroprotective mood-stabilizing agents, its effect on spontaneous brain activity in patients with bipolar depression remains unclear. The aim o...Objective： Although lithium has been a commonly prescribed neurotrophic/neuroprotective mood-stabilizing agents, its effect on spontaneous brain activity in patients with bipolar depression remains unclear. The aim of this study is to reveal the basic mechanism underlying the pathological influences of lithium on resting-state brain function of bipolar depression patients. Methods：97 subjects including 9 bipolar depression patients with lithium treatment, 19 bipolar depression patients without lithium treatment and 69 healthy controls, were recruited to participate in this study. Amplitude of low-frequency fluctuation （ ALFF ） and fractional amplitude of low-frequency fluctuation （ fALFF） were used to capture the changes of spontane-ous brain activity among different groups. In addition, further analysis in terms of Hamilton Depression Rating Scale, the number of depressive episodes, and illness duration in pooled bipolar depression patients were conducted, which combined FLEF and fALEF to identify the basic neural features of bipolar depression patients. Results： It was observed from the imaging results that both the bipolar depression patients receiving lithium treatment and healthy control subjects showed signifi-cantly decreased ALFF/fALFF values in the right anterior cingulate cortex and right middle frontal gyrus compared to that from the bipolar depression patients without lithium treatmetn. The ALFF values of the right＆amp;nbsp;middle temporal gyrus was also found to be negative related to the number of depressive episode and the total episodes. Conclusions：Our findings suggested that the bipolar depression subjects were identified to have ab-normal ALFF/ fALFF in the corticolimbic systems, in-cluding regions like right anterior cingulate cortex, bilateral middle frontal gyrus, right orbital frontal gyrus, and right middle temporal gyrus. In addition, it was also revealed that the decreased ALFF/fALFF in the right anterior cingulate cortex and right middle frontal gyrus might be a biomarker that is related to the lithium effects.展开更多
Acute systemic inflammation critically alters the function of the immune system,often promoting myelopoiesis at the expense of lymphopoiesis.In the thymus,systemic inflammation results in acute thymic atrophy and,cons...Acute systemic inflammation critically alters the function of the immune system,often promoting myelopoiesis at the expense of lymphopoiesis.In the thymus,systemic inflammation results in acute thymic atrophy and,consequently,impaired T-lymphopoiesis.The mechanism by which systemic inflammation impacts the thymus beyond suppressing T-cell development is still unclear.Here,we describe how the synergism between TL1A and IL-18 suppresses T-lymphopoiesis to promote thymic myelopoiesis.The protein levels of these two cytokines were elevated in the thymus during viral-induced thymus atrophy infection with murine cytomegalovirus(MCMV)or pneumonia virus of mice(PVM).In vivo administration of TL1A and IL-18 induced acute thymic atrophy,while thymic neutrophils expanded.Fate mapping with Ms4a3-Cre mice demonstrated that thymic neutrophils emerge from thymic granulocyte-monocyte progenitors(GMPs),while Rag1-Cre fate mapping revealed a common developmental path with lymphocytes.These effects could be modeled ex vivo using neonatal thymic organ cultures(NTOCs),where TL1A and IL-18 synergistically enhanced neutrophil production and egress.NOTCH blockade by the LY411575 inhibitor increased the number of neutrophils in the culture,indicating that NOTCH restricted steady-state thymic granulopoiesis.To promote myelopoiesis,TL1A,and IL-18 synergistically increased GM-CSF levels in the NTOC,which was mainly produced by thymic ILC1s.In support,TL1A-and IL-18-induced granulopoiesis was completely prevented in NTOCs derived from Csf2rb-/-mice and by GM-CSFR antibody blockade,revealing that GM-CSF is the essential factor driving thymic granulopoiesis.Taken together,our findings reveal that TL1A and IL-18 synergism induce acute thymus atrophy while promoting extramedullary thymic granulopoiesis in a NOTCH and GM-CSF-controlled manner.展开更多
The aim of this study is to develop a novel technique for improving the intraoperative margin assessment of glioblastoma by examining the total extrinsic extracellular matrix（ECM） with eosin staining using fluoresce...The aim of this study is to develop a novel technique for improving the intraoperative margin assessment of glioblastoma by examining the total extrinsic extracellular matrix（ECM） with eosin staining using fluorescence lifetime imaging microscopy（FLIM） and scale-invariant feature transform（SIFT） descriptor analysis. Pseudocolor FLIM images obviously exhibit ECM distributions, changes in sequential sections, and different regions of interest. Meanwhile, SIFT descriptors are first utilized for the discrimination of glioblastoma margins by matching similar ECM regions and extracting keypoint orientations from FLIM images obtained from a series of continuous slices. The findings indicate that FLIM imaging with SIFT analysis of the total ECM is a promising method for improving intraoperative diagnosis of frozen and surgically excised brain specimen sections.展开更多
Fluorescence microscopy is the method of choice for studying intracellular dynamics.However,its success depends on the.availability of specific and stable markers.A prominent example of markers that are rapidly gainin...Fluorescence microscopy is the method of choice for studying intracellular dynamics.However,its success depends on the.availability of specific and stable markers.A prominent example of markers that are rapidly gaining interest are nanobodies(Nbs.-15 kDa),which can be functionalized with bright and photostable organic fluorophores.Due to their relatively small size and high specificity,Nbs offer great potential for high-quality long-term subcellular imaging,but suffer from the fact that they cannot spontaneously cross the plasma membrane of live cells.We have recently discovered that laser-induced photoporation is well suited to deliver extrinsic labels to living cells without compromising their viability.Being a laser-based technology,it is readily compatible with light microscopy and the typical cell recipients used for that.Spurred by these promising initial results,we demonstrate here for the first time successful long-term imaging of specific subcellular structures with labeled nanobodies in living cells.We illustrate this using Nbs that target GFP/YFP-protein constructs accessible in the cytoplasm,actin-bundling protein Fascin,and the histone H2A/H2B heterodimers.With an efficiency of more than 80%labeled cells and minimal toxicity(-2%),photoporation proved to be an excellent intracellular delivery method for Nbs.Time-lapse microscopy revealed that cell division rate and migration remained unaffected,confirming excellent cell viability and functionality.We conclude that laser-induced photoporation labeled Nbs can be easily delivered into living cells,laying the foundation for further development of a broad range of Nbs with intracellular targets as a toolbox for long-term live-cell microscopy.展开更多
In the replacement of genetic probes,there is increasing interest in labeling living cells with high-quality extrinsic labels,which avoid over-expression artifacts and are available in a wide spectral range.This calls...In the replacement of genetic probes,there is increasing interest in labeling living cells with high-quality extrinsic labels,which avoid over-expression artifacts and are available in a wide spectral range.This calls for a broadly applicable technology that can deliver such labels unambiguously to the cytosol of living cells.Here,we demonstrate that nanoparticle-sensitized photoporation can be used to this end as an emerging intracellular delivery technique.We replace the traditionally used gold nanoparticles with graphene nanoparticles as photothermal sensitizers to permeabilize the cell membrane upon laser irradiation.We demonstrate that the enhanced thermal stability of graphene quantum dots allows the formation of multiple vapor nanobubbles upon irradiation with short laser pulses,allowing the delivery of a variety of extrinsic cell labels efficiently and homogeneously into live cells.We demonstrate high-quality time-lapse imaging with confocal,total internal reflection fluorescence(TIRF),and Airyscan superresolution microscopy.As the entire procedure is readily compatible with fluorescence(super resolution)microscopy,photoporation with graphene quantum dots has the potential to become the long-awaited generic platform for controlled intracellular delivery of fluorescent labels for live-cell imaging.展开更多
As a hybrid imaging technique, photoacoustic imaging （PAI） can provide multiscale morphological information of tissues, and the use of multi-spectral PAI （MSPAI） can recover the spatial distribution of chromophore...As a hybrid imaging technique, photoacoustic imaging （PAI） can provide multiscale morphological information of tissues, and the use of multi-spectral PAI （MSPAI） can recover the spatial distribution of chromophores of interest, such as hemoglobin within tissues. Herein, we developed a contrast agent that can very effectively combine multiscale PAI with MSPAI for a more comprehensive characterization of complex biological tissues. Specifically, we developed novel PIID-DTBT based semi-conducting polymer dots （Pdots） that show broad and strong optical absorption in the visible-light region （500-700 nm）. The performances of gold nanoparticles （GNPs） and gold nanorods （GNRs）, which have been verified as excellent photoacoustic contrast agents, were compared with that of the Pdots based on the multiscale PAI system. Both ex vivo and in vivo experiments demonstrated that the Pdots have better photoacoustic conversion efficiency at 532 nm than GNPs and showed similar photoacoustic performance with GNRs at 700 nm at the same mass concentration. Photostability and toxicity tests demonstrated that the Pdots are photostable and biocompatible. More importantly, an in vivo MSPAI experiment indicated that the Pdots have better photoacoustic performance than the blood and therefore the signals can be accurately extracted from the background of vascular-rich tissues. Our work demonstrates the great potential of Pdots as highly effective contrast agents for the precise localization of lesions relative to the blood vessels based on multiscale PAI and MSPAI.展开更多
基金funded by the National Natural Science Foundation of China(No.32300659)Shenzhen Science and Technology Innovation Commission Project(No.JCYJ20230807143302004)+6 种基金Zhangjiakou City Key R&D Plan Project(No.2421118D),Zhangjiakou City Key R&D Plan Project(No.2322088D)The Natural Science Project of Hebei North University(No.XJ2024034 and XJ2024035)Medical Science Research Subject Plan Project of Hebei Provincial Health Commission(No.20240782)Project of Administration of Traditional Chinese Medicine of Hebei Province(No.2025392)The 2025 Government-funded Training Project for Outstanding Clinical Medicine Talents(No.ZF2025264)Research Project of Medical Innovation for Chinese Youth(Project Leader:Jun Xue).g Project for Outstanding Clinical Medicine Talents(No.ZF2025264)Research Project of Medical Innovation for Chinese Youth(Project Leader:Jun Xue)。
文摘The structure of intestinal tissue is complex.In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases.Recently,organoids have been successfully constructed and they have come to play an important role in biomedical research.Organoids are miniaturized three-dimensional(3D)organs,derived from stem cells,which mimic the structure,cell types,and physiological functions of an organ,making them robust models for biomedical research.Intestinal organoids are 3D micro-organs derived from intestinal stem cells or pluripotent stem cells that can successfully simulate the complex structure and function of the intestine,thereby providing a valuable platform for intestinal development and disease research.In this article,we review the latest progress in the construction and application of intestinal organoids.
基金Acknowlegements ： The authors gratefully acknowledge the Beijing Normal University Imaging Center for Brain Research and Prof. Yufeng Zang for their contributions in MRI data acquisition. This study was supported by grants from the Beijing Municipal Science ＆ Technology Commission （Grant no. D121100005012002） , Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support （ Grant no. ZYLX201403） , the National Natural Science Foundation of China （ Grant no. 81471389 ） , the High level health technical personnel in Beijing （ Grant no. 2014 - 3-095）, the MYRG2014 - 00093 - FHS and MYRG 2015-00036- FHS grants from the University of Macao in Macao, and FDCT 026/2014/A1 and FDCT 025/ 2015/A1 grants from Macao government.
文摘Objective： Although lithium has been a commonly prescribed neurotrophic/neuroprotective mood-stabilizing agents, its effect on spontaneous brain activity in patients with bipolar depression remains unclear. The aim of this study is to reveal the basic mechanism underlying the pathological influences of lithium on resting-state brain function of bipolar depression patients. Methods：97 subjects including 9 bipolar depression patients with lithium treatment, 19 bipolar depression patients without lithium treatment and 69 healthy controls, were recruited to participate in this study. Amplitude of low-frequency fluctuation （ ALFF ） and fractional amplitude of low-frequency fluctuation （ fALFF） were used to capture the changes of spontane-ous brain activity among different groups. In addition, further analysis in terms of Hamilton Depression Rating Scale, the number of depressive episodes, and illness duration in pooled bipolar depression patients were conducted, which combined FLEF and fALEF to identify the basic neural features of bipolar depression patients. Results： It was observed from the imaging results that both the bipolar depression patients receiving lithium treatment and healthy control subjects showed signifi-cantly decreased ALFF/fALFF values in the right anterior cingulate cortex and right middle frontal gyrus compared to that from the bipolar depression patients without lithium treatmetn. The ALFF values of the right＆amp;nbsp;middle temporal gyrus was also found to be negative related to the number of depressive episode and the total episodes. Conclusions：Our findings suggested that the bipolar depression subjects were identified to have ab-normal ALFF/ fALFF in the corticolimbic systems, in-cluding regions like right anterior cingulate cortex, bilateral middle frontal gyrus, right orbital frontal gyrus, and right middle temporal gyrus. In addition, it was also revealed that the decreased ALFF/fALFF in the right anterior cingulate cortex and right middle frontal gyrus might be a biomarker that is related to the lithium effects.
基金FWO Research Grants G.0B96.20N(PV,PT),G.0C76.18N(PV),G.0B71.18N(PV)G.0A93.22N(PV)+9 种基金Special Research Fund UGent(Methusalem grant BOF16/MET_V/007(PV)BOF22/MET_V/007(PV)iBOF ATLANTIS grant 20/IBF/039(PV))EOS MODEL-IDI Grant(30826052)(PV)EOS CD-INFLADIS(40007512)(PV)Foundation against Cancer(F/2016/865,F/2020/1505)(PV).FWO fundamental research fellowship PhD grant(MRP)(11A7222N)FWO senior postdoctoral fellowship(CM)(12Y2122N)DFG research fellowship(CM)(MA 7770/1-1)Cancer Research Institute Ghent(CRIG)consortiaGhent Gut Inflammation Group(GGIG)consortia.Flanders Institute for Biotechnology(VIB).
文摘Acute systemic inflammation critically alters the function of the immune system,often promoting myelopoiesis at the expense of lymphopoiesis.In the thymus,systemic inflammation results in acute thymic atrophy and,consequently,impaired T-lymphopoiesis.The mechanism by which systemic inflammation impacts the thymus beyond suppressing T-cell development is still unclear.Here,we describe how the synergism between TL1A and IL-18 suppresses T-lymphopoiesis to promote thymic myelopoiesis.The protein levels of these two cytokines were elevated in the thymus during viral-induced thymus atrophy infection with murine cytomegalovirus(MCMV)or pneumonia virus of mice(PVM).In vivo administration of TL1A and IL-18 induced acute thymic atrophy,while thymic neutrophils expanded.Fate mapping with Ms4a3-Cre mice demonstrated that thymic neutrophils emerge from thymic granulocyte-monocyte progenitors(GMPs),while Rag1-Cre fate mapping revealed a common developmental path with lymphocytes.These effects could be modeled ex vivo using neonatal thymic organ cultures(NTOCs),where TL1A and IL-18 synergistically enhanced neutrophil production and egress.NOTCH blockade by the LY411575 inhibitor increased the number of neutrophils in the culture,indicating that NOTCH restricted steady-state thymic granulopoiesis.To promote myelopoiesis,TL1A,and IL-18 synergistically increased GM-CSF levels in the NTOC,which was mainly produced by thymic ILC1s.In support,TL1A-and IL-18-induced granulopoiesis was completely prevented in NTOCs derived from Csf2rb-/-mice and by GM-CSFR antibody blockade,revealing that GM-CSF is the essential factor driving thymic granulopoiesis.Taken together,our findings reveal that TL1A and IL-18 synergism induce acute thymus atrophy while promoting extramedullary thymic granulopoiesis in a NOTCH and GM-CSF-controlled manner.
基金supported by the National Basic Research Program of China(No.2015CB352005)the National Natural Science Foundation of China(Nos.61525503,61378091,and 61620106016)+2 种基金the Guangdong Natural Science Foundation Innovation Team(No.2014A030312008)the Hong Kong,Macao and Taiwan cooperation innovation platform&major projects of international cooperation in Colleges and the Universities in Guangdong Province(No.2015KGJHZ002)the Shenzhen Basic Research Project(Nos.JCYJ20150930104948169,JCYJ2016032814 4746940,and GJHZ20160226202139185)
文摘The aim of this study is to develop a novel technique for improving the intraoperative margin assessment of glioblastoma by examining the total extrinsic extracellular matrix（ECM） with eosin staining using fluorescence lifetime imaging microscopy（FLIM） and scale-invariant feature transform（SIFT） descriptor analysis. Pseudocolor FLIM images obviously exhibit ECM distributions, changes in sequential sections, and different regions of interest. Meanwhile, SIFT descriptors are first utilized for the discrimination of glioblastoma margins by matching similar ECM regions and extracting keypoint orientations from FLIM images obtained from a series of continuous slices. The findings indicate that FLIM imaging with SIFT analysis of the total ECM is a promising method for improving intraoperative diagnosis of frozen and surgically excised brain specimen sections.
基金K.B.acknowledges financial support from the European Research Council(ERC)under the European Union's Horizon 2020 research and innovation program(No.648124)from the Ghent University Special Research Fund(No.01B04912)+3 种基金with gratitude.J.L.gratefully acknowledges the financial support from the China Scholarship Council(CSC)(No.201506750012)the Ghent University Special Research Fund(No.01SC1416)T.H.and J.G.acknowledges financial support from the Fonds Wetenschappelijk Onderzoek(No.G.0559.16N)Ghent University(BOF-GOA)(No.BOF13/GOA010)。
文摘Fluorescence microscopy is the method of choice for studying intracellular dynamics.However,its success depends on the.availability of specific and stable markers.A prominent example of markers that are rapidly gaining interest are nanobodies(Nbs.-15 kDa),which can be functionalized with bright and photostable organic fluorophores.Due to their relatively small size and high specificity,Nbs offer great potential for high-quality long-term subcellular imaging,but suffer from the fact that they cannot spontaneously cross the plasma membrane of live cells.We have recently discovered that laser-induced photoporation is well suited to deliver extrinsic labels to living cells without compromising their viability.Being a laser-based technology,it is readily compatible with light microscopy and the typical cell recipients used for that.Spurred by these promising initial results,we demonstrate here for the first time successful long-term imaging of specific subcellular structures with labeled nanobodies in living cells.We illustrate this using Nbs that target GFP/YFP-protein constructs accessible in the cytoplasm,actin-bundling protein Fascin,and the histone H2A/H2B heterodimers.With an efficiency of more than 80%labeled cells and minimal toxicity(-2%),photoporation proved to be an excellent intracellular delivery method for Nbs.Time-lapse microscopy revealed that cell division rate and migration remained unaffected,confirming excellent cell viability and functionality.We conclude that laser-induced photoporation labeled Nbs can be easily delivered into living cells,laying the foundation for further development of a broad range of Nbs with intracellular targets as a toolbox for long-term live-cell microscopy.
基金support from the European Research Council(ERC)under the European Union’s Horizon 2020 research and innovation program(grant agreement No 648124)from the Ghent University Special Research Fund(01B04912)with gratitude+8 种基金support from the China Scholarship Council(CSC)(201506750012)the Special Research Fund from Ghent University(01SC1416)support from the China Scholarship Council(CSC)(2010634103)from the Research Foundation Flanders(Fonds Wetenschappelijk Onderzoek,FWO)for a doctoral fellowship(11ZB115N)from the Agency for Innovation by Science and Technology(IWT)support from the Centre National de la Recherche Scientifique(CNRS),the University of Lille,the Hauts-de-France region,the CPER“Photonics for Society”the EU union through FLAG-ERA JTC 2015-Graphtivitythe Marie Sklodowska-Curie action(H2020-MSCA-RISE-2015,PANG-690836)support by the FWO Research Community“Scanning and Wide Field Microscopy of(Bio)-organic Systems”and the Province of Limburg(Belgium)for the financial support within the tUL IMPULS FASE II program。
文摘In the replacement of genetic probes,there is increasing interest in labeling living cells with high-quality extrinsic labels,which avoid over-expression artifacts and are available in a wide spectral range.This calls for a broadly applicable technology that can deliver such labels unambiguously to the cytosol of living cells.Here,we demonstrate that nanoparticle-sensitized photoporation can be used to this end as an emerging intracellular delivery technique.We replace the traditionally used gold nanoparticles with graphene nanoparticles as photothermal sensitizers to permeabilize the cell membrane upon laser irradiation.We demonstrate that the enhanced thermal stability of graphene quantum dots allows the formation of multiple vapor nanobubbles upon irradiation with short laser pulses,allowing the delivery of a variety of extrinsic cell labels efficiently and homogeneously into live cells.We demonstrate high-quality time-lapse imaging with confocal,total internal reflection fluorescence(TIRF),and Airyscan superresolution microscopy.As the entire procedure is readily compatible with fluorescence(super resolution)microscopy,photoporation with graphene quantum dots has the potential to become the long-awaited generic platform for controlled intracellular delivery of fluorescent labels for live-cell imaging.
基金Acknowledgements This study was supported by the University of Macao in Macao （Nos. MYRG2014-00093-FHS, MYRG 2015-00036-FHS, and MYRG2016-00110-FHS）, Macao government （Nos. FDCT 026/2014/A1 and FDCT 025/2015/A1）, and the National Natural Science Foundation of China （No. 11434017）.
文摘As a hybrid imaging technique, photoacoustic imaging （PAI） can provide multiscale morphological information of tissues, and the use of multi-spectral PAI （MSPAI） can recover the spatial distribution of chromophores of interest, such as hemoglobin within tissues. Herein, we developed a contrast agent that can very effectively combine multiscale PAI with MSPAI for a more comprehensive characterization of complex biological tissues. Specifically, we developed novel PIID-DTBT based semi-conducting polymer dots （Pdots） that show broad and strong optical absorption in the visible-light region （500-700 nm）. The performances of gold nanoparticles （GNPs） and gold nanorods （GNRs）, which have been verified as excellent photoacoustic contrast agents, were compared with that of the Pdots based on the multiscale PAI system. Both ex vivo and in vivo experiments demonstrated that the Pdots have better photoacoustic conversion efficiency at 532 nm than GNPs and showed similar photoacoustic performance with GNRs at 700 nm at the same mass concentration. Photostability and toxicity tests demonstrated that the Pdots are photostable and biocompatible. More importantly, an in vivo MSPAI experiment indicated that the Pdots have better photoacoustic performance than the blood and therefore the signals can be accurately extracted from the background of vascular-rich tissues. Our work demonstrates the great potential of Pdots as highly effective contrast agents for the precise localization of lesions relative to the blood vessels based on multiscale PAI and MSPAI.
