AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiati...AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.展开更多
Xiaoaiping(XAP)Injection demonstrates the anti-prostate cancer(PCa)effects,yet the underlying mechanism remains unclear.This study aims to investigate the impact of XAP on PCa and elucidate its mechanism of action.PCa...Xiaoaiping(XAP)Injection demonstrates the anti-prostate cancer(PCa)effects,yet the underlying mechanism remains unclear.This study aims to investigate the impact of XAP on PCa and elucidate its mechanism of action.PCa cell proliferation was evaluated using a cell counting kit-8(CCK-8)assay.Cell apoptosis was assessed through Hoechst staining and Western blotting assays.Proteomics technology was employed to identify key molecules and significant signaling pathways modulated by XAP in PCa cells.To further validate potential key genes and important pathways,a series of assays were conducted,including acridine orange(AO)staining,transmission electron microscopy,and immunofluorescence assays.The molecular mechanism of XAP against PCa in vivo was examined using a PC3 xenograft mouse model.Results demonstrated that XAP significantly inhibited cell proliferation in multiple PCa cell lines.In C4-2 and prostate cancer cell line-3(PC3)cells,XAP induced cellular apoptosis,evidenced by reduced B-cell lymphoma 2(Bcl-2)levels and elevated Bcl-2-associated X(Bax)levels.Proteomic,immunofluorescence,and quantitative reverse transcription-polymerase chain reaction(q RT-PCR)investigations revealed a strong correlation between forkhead box O3a(FoxO3a)autophagic degradation and the anti-PCa action of XAP.XAP hindered autophagy by reducing the expression levels of autophagy-related protein 5(Atg5)/autophagy-related protein 12(Atg12)and enhancing FoxO3a expression and nuclear translocation.Furthermore,XAP exhibited potent anti-PCa action in PC3 xenograft mice and triggered FoxO3a nuclear translocation in tumor tissue.These findings suggest that XAP induces PCa apoptosis via inhibition of FoxO3a autophagic degradation,potentially offering a novel perspective on XAP injection as an effective anticancer therapy for PCa.展开更多
AIM:To investigate the role of Forkhead box protein P3(FOXP3)in choroidal melanoma(CM)metastases and elucidate its underlying mechanisms.METHODS:FOXP3 protein expression was analyzed in CM clinical specimens and cell ...AIM:To investigate the role of Forkhead box protein P3(FOXP3)in choroidal melanoma(CM)metastases and elucidate its underlying mechanisms.METHODS:FOXP3 protein expression was analyzed in CM clinical specimens and cell lines.A stable FOXP3 knockout cell line and a transient FOXP3-overexpressing cell line were established,with transfection efficiencies confirmed by Western blotting(WB).Functional assays,including monoclonal formation,cell counting kit-8(CCK-8)proliferation,migration,invasion,and in vivo tumorigenesis assays in nude mice,were performed to assess the biological effects of FOXP3.Additionally,WB was employed to evaluate epithelial-mesenchymal transition(EMT)markers and the activation of the Wnt5a/CaMKII signaling pathway.RESULTS:FOXP3 expression was significantly elevated in both CM clinical specimens and cell lines.Functional analyses revealed that FOXP3 enhanced CM cell proliferation,migration,and invasion in vitro and promoted tumorigenesis in vivo.Mechanistically,FOXP3 upregulated EMT-related proteins and activated the Wnt5a/CaMKII signaling pathway.Rescue experiments further confirmed that the oncogenic effects of FOXP3 were mediated via modulation of the Wnt5a/CaMKII axis.CONCLUSION:This study identifies FOXP3 as an oncogenic driver in CM,promoting tumor progression through the Wnt5a/CaMKII signaling pathway.These findings provide new insights into the molecular mechanisms of CM pathogenesis and highlight FOXP3 as a potential therapeutic target.展开更多
Precise and robust three-dimensional object detection(3DOD)presents a promising opportunity in the field of mobile robot(MR)navigation.Monocular 3DOD techniques typically involve extending existing twodimensional obje...Precise and robust three-dimensional object detection(3DOD)presents a promising opportunity in the field of mobile robot(MR)navigation.Monocular 3DOD techniques typically involve extending existing twodimensional object detection(2DOD)frameworks to predict the three-dimensional bounding box(3DBB)of objects captured in 2D RGB images.However,these methods often require multiple images,making them less feasible for various real-time scenarios.To address these challenges,the emergence of agile convolutional neural networks(CNNs)capable of inferring depth froma single image opens a new avenue for investigation.The paper proposes a novel ELDENet network designed to produce cost-effective 3DBounding Box Estimation(3D-BBE)froma single image.This novel framework comprises the PP-LCNet as the encoder and a fast convolutional decoder.Additionally,this integration includes a Squeeze-Exploit(SE)module utilizing the Math Kernel Library for Deep Neural Networks(MKLDNN)optimizer to enhance convolutional efficiency and streamline model size during effective training.Meanwhile,the proposed multi-scale sub-pixel decoder generates high-quality depth maps while maintaining a compact structure.Furthermore,the generated depthmaps provide a clear perspective with distance details of objects in the environment.These depth insights are combined with 2DOD for precise evaluation of 3D Bounding Boxes(3DBB),facilitating scene understanding and optimal route planning for mobile robots.Based on the estimated object center of the 3DBB,the Deep Reinforcement Learning(DRL)-based obstacle avoidance strategy for MRs is developed.Experimental results demonstrate that our model achieves state-of-the-art performance across three datasets:NYU-V2,KITTI,and Cityscapes.Overall,this framework shows significant potential for adaptation in intelligent mechatronic systems,particularly in developing knowledge-driven systems for mobile robot navigation.展开更多
基金Supported by the National Natural Science Foundation of China(No.81800423)the Guangdong Medical Science and Technology Research project(No.B2022102)。
文摘AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.
基金supported by Guangxi Natural Science Foundation(No.2023GXNSFDA026047)Guangxi Science and Technology Project(No.FANGKE ZY20221502)+1 种基金the National Natural Science Foundation of China(No.82160948)the Advanced Innovation Teams and Xinghu Scholars Program of Guangxi Medical University。
文摘Xiaoaiping(XAP)Injection demonstrates the anti-prostate cancer(PCa)effects,yet the underlying mechanism remains unclear.This study aims to investigate the impact of XAP on PCa and elucidate its mechanism of action.PCa cell proliferation was evaluated using a cell counting kit-8(CCK-8)assay.Cell apoptosis was assessed through Hoechst staining and Western blotting assays.Proteomics technology was employed to identify key molecules and significant signaling pathways modulated by XAP in PCa cells.To further validate potential key genes and important pathways,a series of assays were conducted,including acridine orange(AO)staining,transmission electron microscopy,and immunofluorescence assays.The molecular mechanism of XAP against PCa in vivo was examined using a PC3 xenograft mouse model.Results demonstrated that XAP significantly inhibited cell proliferation in multiple PCa cell lines.In C4-2 and prostate cancer cell line-3(PC3)cells,XAP induced cellular apoptosis,evidenced by reduced B-cell lymphoma 2(Bcl-2)levels and elevated Bcl-2-associated X(Bax)levels.Proteomic,immunofluorescence,and quantitative reverse transcription-polymerase chain reaction(q RT-PCR)investigations revealed a strong correlation between forkhead box O3a(FoxO3a)autophagic degradation and the anti-PCa action of XAP.XAP hindered autophagy by reducing the expression levels of autophagy-related protein 5(Atg5)/autophagy-related protein 12(Atg12)and enhancing FoxO3a expression and nuclear translocation.Furthermore,XAP exhibited potent anti-PCa action in PC3 xenograft mice and triggered FoxO3a nuclear translocation in tumor tissue.These findings suggest that XAP induces PCa apoptosis via inhibition of FoxO3a autophagic degradation,potentially offering a novel perspective on XAP injection as an effective anticancer therapy for PCa.
基金Supported by the National Natural Science Foundation of China(No.81873345,No.82274585)Qingdao Key Health Discipline Development Fund.
文摘AIM:To investigate the role of Forkhead box protein P3(FOXP3)in choroidal melanoma(CM)metastases and elucidate its underlying mechanisms.METHODS:FOXP3 protein expression was analyzed in CM clinical specimens and cell lines.A stable FOXP3 knockout cell line and a transient FOXP3-overexpressing cell line were established,with transfection efficiencies confirmed by Western blotting(WB).Functional assays,including monoclonal formation,cell counting kit-8(CCK-8)proliferation,migration,invasion,and in vivo tumorigenesis assays in nude mice,were performed to assess the biological effects of FOXP3.Additionally,WB was employed to evaluate epithelial-mesenchymal transition(EMT)markers and the activation of the Wnt5a/CaMKII signaling pathway.RESULTS:FOXP3 expression was significantly elevated in both CM clinical specimens and cell lines.Functional analyses revealed that FOXP3 enhanced CM cell proliferation,migration,and invasion in vitro and promoted tumorigenesis in vivo.Mechanistically,FOXP3 upregulated EMT-related proteins and activated the Wnt5a/CaMKII signaling pathway.Rescue experiments further confirmed that the oncogenic effects of FOXP3 were mediated via modulation of the Wnt5a/CaMKII axis.CONCLUSION:This study identifies FOXP3 as an oncogenic driver in CM,promoting tumor progression through the Wnt5a/CaMKII signaling pathway.These findings provide new insights into the molecular mechanisms of CM pathogenesis and highlight FOXP3 as a potential therapeutic target.
文摘Precise and robust three-dimensional object detection(3DOD)presents a promising opportunity in the field of mobile robot(MR)navigation.Monocular 3DOD techniques typically involve extending existing twodimensional object detection(2DOD)frameworks to predict the three-dimensional bounding box(3DBB)of objects captured in 2D RGB images.However,these methods often require multiple images,making them less feasible for various real-time scenarios.To address these challenges,the emergence of agile convolutional neural networks(CNNs)capable of inferring depth froma single image opens a new avenue for investigation.The paper proposes a novel ELDENet network designed to produce cost-effective 3DBounding Box Estimation(3D-BBE)froma single image.This novel framework comprises the PP-LCNet as the encoder and a fast convolutional decoder.Additionally,this integration includes a Squeeze-Exploit(SE)module utilizing the Math Kernel Library for Deep Neural Networks(MKLDNN)optimizer to enhance convolutional efficiency and streamline model size during effective training.Meanwhile,the proposed multi-scale sub-pixel decoder generates high-quality depth maps while maintaining a compact structure.Furthermore,the generated depthmaps provide a clear perspective with distance details of objects in the environment.These depth insights are combined with 2DOD for precise evaluation of 3D Bounding Boxes(3DBB),facilitating scene understanding and optimal route planning for mobile robots.Based on the estimated object center of the 3DBB,the Deep Reinforcement Learning(DRL)-based obstacle avoidance strategy for MRs is developed.Experimental results demonstrate that our model achieves state-of-the-art performance across three datasets:NYU-V2,KITTI,and Cityscapes.Overall,this framework shows significant potential for adaptation in intelligent mechatronic systems,particularly in developing knowledge-driven systems for mobile robot navigation.
