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(A) Schematic illustration of the therapeutic strategy for melanoma based on HA@BCN. (B) Live/dead staining of B16F10 cells in different groups. (C) Growth of tumor volume in different treatment groups. (D) Digital photographs of skin regeneration during the repair period in different groups. (Adapted with permission from Zhao et al., 2022. Copyright 2022 American Chemical Society). (E) Micro-CT images and value of BV/TV, BMS, and TOT at 24 weeks after implantation in different groups. (Adapted with permission from Pan et al., 2020. Copyright 2019 Wiley-VCH).

PMC10020212

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(A) Schematic illustration of the therapeutic strategy for bacterial infection based on Nb2C@TP. (B) SEM images of biofilm-resistance performance in different groups. (C) TEM images of antibacterial performance in different groups. (Adapted with permission from Yang et al., 2021. Copyright 2021 American Chemical Society).

PMC10020212

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(A) Schematic illustration of release and activation of SN38-Nif based on ZrC@prodrug. (B) Drug release percentage with and without irradiation. (C) Relative cell viability of SMMC-7721 in different groups. (D) Growth of tumor volume in different treatment groups. (E) Digital photographs of tumor after treatments in different groups. (Adapted with permission from Liu S et al., 2020. Copyright 2020 Wiley-VCH).

PMC10020212

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(A) Schematic illustration of fabrication of FePSe3@APP@CCM and the strategy of combined PTT immunotherapy. (B) Schematic illustration of DC maturation during the treatment. (C) Flow cytometry analysis of DC maturation in different groups. (Adapted with permission from Fang et al., 2021. Copyright 2020 Wiley-VCH).

PMC10020212

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(A) SEM images of the interface between Ti3C2TX films and DRG neurons. (B) SEM images of the interface between Ti3C2TX flakes and DRG neurons. (C) Illustration of Ti3C2TX film-mediated DRG electrical activity modulation. (D) Illustration of Ti3C2TX flake-mediated DRG electrical activity modulation. (E) Illustration of selective modulation of DRG neurons mediated by Ti3C2TX films. (F) Illustration of selective modulation of DRG neurons mediated by Ti3C2TX flakes. (Adapted with permission from Wang et al., 2021a. Copyright 2021 American Chemical Society).

PMC10020212

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(A) Schematic illustration of the therapeutic strategy for IUA prevention based on CUP/PT. (B) Photothermal property of PT and CUP/PT under 1064-nm laser irradiation. (C) Shape recovery of CUP/PT at different time after irradiation on a desk (I), underneath the skin (II), and inside the isolated uterine lumen of rat (III), respectively. (Adapted with permission from Dong et al., 2022. Copyright 2022 Wiley-VCH).

PMC10020212

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(A) Schematic illustration of photothermal-mediated cellular uptake of BP-HAS-PTX. (B) CLSM images of U87MG cells incubated with PBS (I), BP-HAS-PTX without laser irradiation (II), and BP-HAS-PTX with laser irradiation (III), respectively. (C) Mean FITC fluorescence intensity of U87MG cells in different groups. (Adapted with permission from Wang et al., 2017. Copyright 2017 Ivyspring International Publisher).

PMC10020212

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(A) Schematic illustration of fabrication of Ti3C2-DOX and therapeutic strategy of based on Ti3C2-DOX. (B) Absorbance of DCPF in different groups. (C) CLSM images of HCT-116c cells incubated with Ti3C2-DOX after irradiation. (Adapted with permission from Liu et al., 2017. Copyright 2017 American Chemical Society). (D) Schematic illustration of therapeutic strategy for bacterial infection and wound healing based on CS-BP. (E) ESP spectra in different groups. (F) Images of bacterial colony in different groups after treatments. (Adapted with permission from Mao et al., 2018. Copyright 2018 American Chemical Society).

PMC10020212

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Hypoxia induces ULK1-related autophagy and mitochondrial Lon in cancer cells.A HCT-15 cells were exposed to hypoxia (1% O2) for 24 h or CoCl2 (200 µM) for 16 h and the collected lysates were immunoblotted against the mitophagy signaling using indicated antibodies. GAPDH as the loading control. B HCT-15 cells were exposed to hypoxia (1% O2) for the indicated time points and the protein levels of ULK1 signaling and LC3B were obtained after immunoblotting. Cell lysates were analyzed by immunoblotting using indicated antibodies. GAPDH as the loading control. C HCT-15 cells were treated with CoCl2 (200 μM) for different time points (0–48 h). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. D, E Hypoxia mimic induces lysosome-mediated autophagy. D Hypoxia mimic-induced lysosome-mediated autophagy were verified by immunofluorescence. HCT-15 cells were treated with or without CoCl2 (200 μM for 18 h) and with BafilomycinA1 (100 nM for 6 h) or not. The cells were fixed and immunostained by LC3B (green) and anti-LAMP1 (red) antibodies. DNA was stained with DAPI (blue). Scale bars = 50 μm. E Hypoxia mimic-induced lysosome-mediated autophagy were quantified by LC3B puncta/cell detection according to the immunofluorescence data in D. LC3B puncta/cell was quantified by selecting more than 50 cells per condition. n = 3 biological replicates. The error bars shown in the panel represent the standard deviation from three independent experiments. ***p < 0.001. F, G Hypoxia mimic induces lysosome-mediated mitophagy. F Hypoxia mimic-induced lysosome-mediated mitophagy were verified by confocal immunofluorescence. HCT-15 cells transiently expressing mito-QC were treated with or without CoCl2 (200 μM for 18 h) and with BafilomycinA1 (100 nM for 6 h) or not. The cells were fixed and immunostained by GFP (mitochondria, green), mCherry (mitolysosome, red), and anti-LAMP1 (lysosome, blue) antibodies. Scale bars = 10 μm. G Hypoxia mimic-induced lysosome-mediated mitophagy were quantified by mcherry signals according to the immunofluorescence data in F. n = 3 biological replicates. The error bars shown in the panel represent the standard deviation from three independent experiments. ***p < 0.001.

PMC10020552

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Mitochondrial Lon is required for ULK1-mediated mitophagy under hypoxia.A, B HCT-15 cells were transiently transfected with the plasmids encoding Lon or Lon-shRNA. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. C HCT-15 cells transfected with the plasmids encoding Lon or empty were treated with SBI-0206965 (20 μM for 6 h) or not. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. D HCT-15 cells were treated with CoCl2 (200 μM for 18 h) or not in the presence or absence of SBI-0206965 (20 μM for 6 h). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. E, F Mitochondrial Lon induces lysosome-mediated autophagy. E Mitochondrial Lon-induced lysosome-mediated autophagy were verified by immunofluorescence. HCT-15 cells were transfected with the plasmids encoding Lon or empty and treated with BafilomycinA1 (100 nM for 6 h) or not. The cells were fixed and immunostained by RFP-LC3B (red) and anti-LAMP1 (green) antibodies. Scale bars = 50 μm. F Mitochondrial Lon-induced lysosome-mediated autophagy were quantified by LC3B puncta/cell detection according to the immunofluorescence data in E. LC3B puncta/cell was quantified by selecting more than 50 cells per condition. n = 3 biological replicates. The error bars shown in the panel represent the standard deviation from three independent experiments. ***p < 0.001. G, H Mitochondrial Lon induces lysosome-mediated mitophagy. G Mitochondrial Lon-induced lysosome-mediated mitophagy were verified by confocal immunofluorescence. HCT-15 cells transiently expressing mito-QC were transfected with the plasmids encoding Lon or empty and treated with BafilomycinA1 (100 nM for 6 h) or not. The cells were fixed and immunostained by GFP (mitochondria, green), mCherry (mitolysosome, red), and anti-LAMP1 (lysosome, blue) antibodies. Scale bars = 10 μm. H Mitochondrial Lon-induced lysosome-mediated mitophagy were quantified by mcherry signals according to the immunofluorescence data in F. n = 3 biological replicates. The error bars shown in the panel represent the standard deviation from three independent experiments. ***p < 0.001.

PMC10020552

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Mitochondrial Lon chaperone activity contributes to the stability of ULK1 complex for the mitophagy activation.A HCT-15 cells were transfected with the plasmids encoding pcDNA3-Lon in different concentrations (0.5–5 μg). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. B HCT-15 cells were transfected with the plasmids encoding myc-Lon, myc-LonK529R (ATPase mutant), or myc-LonS855A (protease mutant). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. C, D HCT-15 cells were transfected with the plasmids encoding empty, myc-Lon, or myc-LonK529R in the presence or absence of CoCl2 treatment (200 μM for 18 h). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. E HCT-15 cells transfected with the plasmids encoding Lon-shRNA or empty were treated with or without MG132 (10 μM for 6 h). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. F HCT-15 cells transfected with the plasmids encoding Lon-shRNA or empty vector were treated with or without Cycloheximide (50 µg/mL) for the indicated time course. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control.

PMC10020552

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Lon and ULK1 complex accumulates at ER-mitochondria tethering sites in response to hypoxia.A, B Lon and ULK1 complex accumulates at ER-mitochondria contact (EMC) sites under hypoxia. HCT-15 cells treated with or without CoCl2 (200 μM for 18 h) (A) or transfected with the plasmids encoding myc-Lon (B) were used to perform subcellular fractionation experiment. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. Mito mitochondria, MAM mitochondria associated membranes, ER endoplasmic reticulum, cyto cytosol, PNS post nuclear supernatant. C–E Accumulation of Lon and ULK1 at the EMC/MAM was verified by confocal immunofluorescence. HCT-15 cells were transfected with the plasmids encoding Lon or treated with CoCl2 (200 μM for 18 h) or not. The cells were fixed and immunostained by anti-Lon (green) (C), anti-ULK1 (green) (D), anti-FACL4 (ER and MAM, green) (E), anti-SERCA-2 (ER, blue), and anti-TOMM20 (mitochondria, red) antibodies. Scale bars = 10 μm. F–H Localizaton of Lon at the EMC/MAM was verified by transmission electron microscopy (TEM). HCT-15 cells were treated with CoCl2 (200 μM for 18 h). The cells were fixed and and immunostained by anti-Lon and immunogold labeling antibodies. The immunogold electron micrographs showed Lon in (i) damaged mitochondria (M), (ii) ER around Nucleus (N), (iii) Cytosol (F) and the ER-mitochondria tethering sites (G). ER: endoplasmic reticulum. Scale bars: 100 nm. Quantitation of the percentage of ER adjacent to mitochondria in both CoCl2 and Lon expressed HCT-15 cells and compared with control (H). The percentage was normalized by total ER and mitochondrial perimeter (n = 36 field for each condition). n = 3 biological replicates. The error bars shown in the panel represent the standard deviation from three independent experiments. ***p < 0.001.

PMC10020552

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Mitochondrial Lon interacts with and stabilizes ULK1 complex under hypoxia.A–D Mitochondrial Lon interacts with ULK1 shown by co-immunoprecipitation. A HCT-15 cells were treated with CoCl2 followed by co-immunoprecipitation with anti-ULK1. Whole cell lysates from HCT-15 cells treated with CoCl2 (200 μM for 18 h) were immunoprecipitated with anti-ULK1 antibodies. The immunoprecipitation complex was analyzed by Western blotting using the indicated antibodies. IP, immunoprecipitation. B Whole cell lysates from HCT-15 cells transfected with the plasmids encoding Flag-ULK1 or vector were immunoprecipitated with anti-ULK1 antibodies. The immunoprecipitation complex was analyzed by Western blotting using the indicated antibodies. C Whole cell lysates from HCT-15 cells transfected with the plasmids encoding myc-Lon and Flag-ULK1 were immunoprecipitated with anti-myc antibodies. The immunoprecipitation complex was analyzed by Western blotting using the indicated antibodies. D Whole cell lysates from HCT-15 cells transfected with the plasmids encoding myc-Lon or myc-LonK529R were immunoprecipitated with anti-myc antibodies. The immunoprecipitation complex was analyzed by Western blotting using the indicated antibodies. E, F Mitochondrial Lon interacts with ULK1 shown by immunofluorescence. E The interaction of Lon with ULK1 was enhanced by ULK1 activity under hypoxia. HCT-15 cells treated with or without CoCl2 (200 μM for 18 h) in the presence or absence of SBI-0206965 (20 μM for 6 h) were immunostained by anti-ULK1 (green) and anti-Lon (red) following image capturing by immunofluorescence microscopy. DAPI was used for nuclear staining. Scale bars, 50 μm (n = >50 cells/condition and 3 biological replicates). F The interaction of Lon with ULK1 was enhanced by ULK1 activity. HCT-15 cells transfected with the plasmids encoding Lon or empty in the presence or absence of SBI-0206965 (20 μM for 6 h) were immunostained by anti-ULK1 (green) and anti-Lon (red) following image capturing by immunofluorescence microscopy. DAPI was used for nuclear staining. Scale bars, 50 μm (n = >50 cells/condition and 3 biological replicates).

PMC10020552

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FUNDC1-Ser17 phosphorylation by ULK1 kinase at the EMC/MAM is important for the Lon-induced mitophagy.A HCT-15 cells transfected with the plasmids encoding myc-Lon were used to perform subcellular fractionation experiment. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. Mito mitochondria, MAM mitochondria associated membranes, ER endoplasmic reticulum, cyto cytosol, PNS post nuclear supernatant. B HCT-15 cells were treated with CoCl2 (200 μM for 18 h) or not in the presence or absence of SBI-0206965 (20 μM for 6 h). Whole cell lysates from the treated HCT-15 cells were immunoprecipitated with anti-LC3B antibodies. The immunoprecipitation complex was analyzed by Western blotting using the indicated antibodies. C HCT-15 cells transfected with the plasmids encoding Lon or empty vector were treated with SBI-0206965 (20 μM for 6 h) or not. Whole cell lysates from the treated HCT-15 cells were immunoprecipitated with anti-LC3B antibodies. The immunoprecipitation complex was analyzed by Western blotting using the indicated antibodies. D HCT-15 cells were treated with or without CoCl2 (200 μM for 18 h) or hypoxia exposure (1% O2). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. E HCT-15 cells were transfected with the plasmids encoding Lon or Lon-shRNA. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. F HCT-15 cells transfected with the plasmids encoding Lon or empty were treated with SBI-0206965 (20 μM for 6 h) or not. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. G HCT-15 cells were transfected with the plasmids encoding myc-Lon, myc-LonK529R (ATPase mutant), or myc-LonS855A (protease mutant). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. H HCT-15 cells were transfected with the plasmids encoding empty, myc-Lon, or myc-LonK529R in the presence or absence of CoCl2 treatment (200 μM for 18 h). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. I Immunohistochemical analysis of p-FUNDC1-S17 expression in OSCC patients. Representative immunohistochemical staining of p-FUNDC1-S17 was performed using paraffin-embedded sections of OSCC. The representative intensity of immunostaining was classified as four levels: negative staining intensity (0) and positive staining intensity, including weak (1+), median (2+), and strong (3+) staining. Microscopic magnification, ×400. Scale bar, 50 μm.

PMC10020552

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Lon binds with mitochondrial Na+/Ca2+ exchanger to promote FUNDC1-ULK-mediated mitophagy in the EMC/MAM site.A, B Ca2+mito (mt-lar-GECO) and Ca2+cyto (Fura-2 AM) were measured by live-cell microscopy in OEC-M1 treated with CoCl2 (300 µM-18h) or transfected with Lon in presence or absence of CGP37157 (10 µM-4h). ATP (100 µM) was used as an agonist and further measured and analyzed (n = 3). C OEC-M1 cells were treated with CoCl2 (300 µM-18 h) in the presence or absence of CGP37157 (10 µM-4 h). Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. D OEC-M1 cells were transfected with Vector, Lon, NCLX (48 h) in the presence or absence of CGP37157 (10 µM-4h) and shNCLX transfected OEC-M1 cells were co-transfected with NCLX and Lon plasmids and incubated for 48 h. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. E, F Lon and ULK1 complex accumulation at EMC/MAM sites were abolished upon NCLX inhibition under hypoxia. OEC-M1 cells transfected with the plasmids encoding myc-Lon (E) or treated with CoCl2 (200 μM for 18 h) (F) in presence or absence of CGP37157 (10 µM-8h) were used to perform subcellular fractionation experiments. Cell lysates were analyzed by immunoblotting using the indicated antibodies. GAPDH as the loading control. Mito mitochondria, EMC ER-mitochondria contact sites, MAM mitochondria associated membranes, WCL whole cell lysate.

PMC10020552

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Lon-ROS-ULK1-FUNDC1 axis induced mitophagy benefits cell survival and tumorigenesis in vitro and in vivo.A–C HCT-15 cells were treated with CoCl2 (200 μM for 18 h) or not in the presence or absence of SBI-0206965 (20 μM for 24 h). The MTS assay for cell viability (A), fluorescence-based cleaved Caspase 3 apoptosis assay (B), and western blotting analysis (C) were performed. Immunoblots were obtained using the indicated antibodies. Scale bar, 100 μm (n = >50 cells/condition and 3 biological replicates). D–F HCT-15 cells were treated with CoCl2 (200 μM for 18 h) or not in the presence or absence of Bafilomycin A1 (100 nM for 24 h). The MTS assay for cell viability (D), fluorescence-based cleaved Caspase 3 apoptosis assay (E), and Western blotting analysis (F) were performed. Immunoblots were obtained using the indicated antibodies. Scale bar, 100 μm (n = >50 cells/condition and 3 biological replicates). G Immunohistochemical analysis of ULK1, p-FUNDC1 S17, FUNDC1, LC3B, and Bcl-2 expression in Lon expressed OEC-M1 tumor generated in BALB/C Nu mice treated with or without CGP37157. Representative immunohistochemical staining of respective targets was performed using paraffin-embedded sections of tumors collected. Microscopic magnification, ×400. Scale bar, 50 μm. H Scheme of the interaction between mitochondrial Lon and ULK1 complex at the EMC/MAM promotes mitophagy under hypoxia by stabilizing FUNDC1-ULK1 complex that depends on mitochondrial Na+/Ca2+ exchanger (NCLX). Upon hypoxia, Lon promotes FUNDC1-ULK1-mediated mitophagy at the EMC/MAM site, which is dependent on the binding with mitochondrial Na+/Ca2+ exchanger (NCLX). This interaction stabilized the FUNDC1-ULK1 at the EMC and initiated the mitophagy through the regulation of Ca2+ levels between mitochondria and cytosol. Lon-ULK1 phosphorylates FUNDC1 at S17, and Lon-ULK1-FUNDC1 axis promotes mitophagosome-lysosome fusion. EMC ER-mitochondria contact sites, MAM mitochondria associated membranes. The scheme of this study was created with BioRender.com.

PMC10020552

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Neuropathic pain developed in SCI mice. (A) The SCI mouse model was generated by punching a hole in left L5 segment of spinal cord under vertebra thoracic segment 13 (T13) using a 26G needle. Injury site is marked with a red circle. (B) Hematoxylin and Eosin (H&E) staining of spinal cord sections showed that the injury track only presented in SCI mice, not in naive nor sham mice. (C) The paw-withdrawal threshold of the ipsilateral side of sham mice pre-surgery and three, five, and eight days post operation (DPO), n = 6. (D) The paw-withdrawal threshold of the ipsilateral side of SCI mice significantly dropped at 3 DPO and further decreased by 8 DPO. Baseline 1.3 ± 0.30 g; Day 3, 0.8 ± 0.35 g; Day 5, 0.7 ± 0.28 g; Day 8, 0.3 ± 0.12 g, n = 7. * p < 0.05, ** p < 0.01, **** p < 0.0001, unpaired t-test or one-way ANOVA. n.s. not significant. Scale bar = 100 μm.

PMC10020601

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Upregulation of Nav1.7 in DRG and SDH of SCI mice. (A) The expression level of Nav1.7 in DRG and SDH of naive, sham, and SCI mice was measured by Western blot. (B) Quantitative analysis of duplicate Western-blot membranes showed that compared to naive mice, the expression level of Nav1.7 increased significantly in DRG of SCI mice (1.3 ± 0.10), and an increasing trend in expression level of Nav1.7 was also observed in sham mice (1.1 ± 0.19). The expression level of Nav1.7 in SDH of SCI and sham mice was 2.2-fold and 1.5-fold higher than that in naive mice, respectively, (2.2 ± 1.50, 1.5 ± 1.03). (C) Immunostaining for Nav1.7 (Green) on spinal section derived from naive, sham, and SCI mice. Arrows point to Nav1.7 positive neurons, DAPI is counterstaining for nuclei. Inset is the high magnification view of the boxed area. (D) The number of neurons expressing Nav1.7 in laminae I-VI of naive, sham, and SCI mice. # represents the comparison between SCI and sham，* represents the comparison between naïve and SCI or sham. (E) Immunostaining for Nav1.7 (Green) on DRG section derived from naive, sham, and SCI mice demonstrated the upregulation of Nav1.7 in DRG neurons in SCI and sham mice when compared to that in naive mice. Inset is the high magnification view of the boxed area. Data are shown as Mean ± SD, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, ## p < 0.01, ### p < 0.001, unpaired t-test or one way ANOVA. n.s. not significant. Scale bar = 100 μm.

PMC10020601

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Induced expression of NGF in DRG and SDH of mice after SCI. (A) The expression level of NGF in DRG and SDH of naive, sham, and SCI mice was measured by Western blot. (B) Quantitative analysis of duplicate Western-blot membranes showed that there was an increase trend in the expression level of NGF in DRG and SDH in SCI and sham mice when compared to naive mice (sham DRG 1.5 ± 0.34, SCI DRG 1.7 ± 0.52; sham SDH 1.1 ± 0.11, SCI SDH 1.2 ± 0.32). (C) Immunostaining for NGF (Green) on spinal cord section derived from naive, sham, and SCI mice. Arrows point to NGF positive neurons. Inset is the high magnification view of the boxed area. (D) The number of neurons expressing NGF in laminae I-VI of naive, sham, and SCI. # represents the comparison between SCI and sham, * represents the comparison between naive and SCI or sham. (E, F) Immunostaining for NGF (Green) on DRG section derived from naive, sham, and SCI mice (E) and the number of neurons expressing high levels of NGF in DRG neurons in naive, sham, and SCI mice. Inset is the high magnification view of the boxed area. Data are shown as Mean ± SD, n = 3. ## p < 0.01, ###p < 0.001, *p < 0.05, **p < 0.01, ***p < 0.001,unpaired t-test or one way ANOVA. n.s. not significant. Scale bar = 100 μm.

PMC10020601

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Expression of voltage-gated sodium channels in spinal dorsal horn and DRG of SCI mice at 12 DPO. (A) Western blot showed that the expression level of Nav1.2 was not significantly upregulated in SDH of SCI mice when compared to that in naive mice. (B) Western blot demonstrated that there was no significant difference in the expression level of Nav1.8 in SDH between SCI mice and naive mice. (C) The expression level of Nav1.7 in SDH of naive and SCI mice measured by Western-blot assay. (D) The expression level of Nav1.7 in DRG significantly increased 1.5-fold (1.5 ± 0.32) in SCI mice compared to that in naive mice. Data are shown as Mean ± SD, * p < 0.05, n = 4, unpaired t-test. n.s. not significant. Scale bar = 100 μm.

PMC10020601

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Blockers of Nav1.7-alleviated mechanical pain in SCI mice. (A) Nav1.7 blocker PF-05089771 significantly relieved mechanical pain of SCI mice at doses of both 2 mg/kg and 4 mg/kg (Vehicle 0.03 ± 0.08 g, 2 mg/kg 30 min 0.2 ± 0.13 g, 4 mg/kg 30 min 0.3 ± 0.16 g, 2 mg/kg 60 min 0.4 ± 0.19 g, and 4 mg/kg 60 min 0.4 ± 0.20) g. △PWT = post-drug PWT - pre-drug PWT. (B) Nav1.7 blocker GNE-0439 significantly alleviated mechanical pain of SCI mice at doses from 10 μg/kg to 30 μg/kg (10 μg/kg 30 min 0.3 ± 0.15 g, 20 μg/kg 30 min 0.5 ± 0.28 g, 30 μg/kg 30 min 0.2 ± 0.11 g, 10 μg/kg 60 min 0.2 ± 0.18 g, 20 μg/kg 60 min 0.3 ± 0.20 g, and 30 μg/kg 60 min 0.2 ± 0.08 g). (C) Gabapentin significantly reduced mechanical pain of SCI mice at dose of 50 mg/kg (30 min 0.5 ± 0.36 g and 60 min 0.5 ± 0.30 g). (D) The efficacy of GNE-0439 to relieve mechanical pain was equivalent to Gabapentin, and the efficacy of GNE-0439 and Gabapentin was slightly better than PF-05089771, but not statistically significant (PF-05089771 0.4 ± 0.20 g, GNE-0439 0.5 ± 0.28 g, and Gabapentin 0.5 ± 0.36 g). (E) Response ratio of paw withdrawal of SCI mice administrated neither PF-05089771 (blue line) or GNE-0439 (red line) to stimulus from 0.07 g Von Frey monofilament to 1.0 g Von Frey monofilament. Data are shown as Mean ± SD, n = 7. *p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, unpaired t-test or one-way ANOVA, or two-way ANOVA. n.s. not significant.

PMC10020601

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Activation of Nav1.7+ SDH neurons in SCI mice. (A–L) Double immunostaining for FOS (Red) and Nav1.7 (Green) on spinal section derived from naive (A, D, G, J), sham (B, E, H, K), and SCI (C, F, I, L) mice showed more FOS-positive neurons and more FOS/Nav1.7 double-positive neurons in deep laminae layers of SCI mice. Inset is the high magnification view of the boxed area. Scale bar 100 μm. (M–O) The number of FOS+ (M), Nav1.7+/FOS+ (N), and Nav1.7+ (O) SDH neurons in laminae I–VI of home-caged naive mice, sham, and SCI mice. The symbol # represents the comparison between SCI and sham, and the symbol * represents the comparison between naive and SCI or sham. (P) The ratio of Nav1.7+/FOS+ SDH neurons to total Nav1.7+ SDH neurons in naive (0%), sham (3 ± 4.8%), and SCI (26 ± 8.2%) mice. Data are shown as Mean ± SD, n = 3. # p < 0.05, ## p < 0.01,*p < 0.05, ** p < 0.01, ***p < 0.001, unpaired t-test or one way ANOVA. Scale bar = 100 μm.

PMC10020601

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Increase of phosphorylated JUN in SDH and DRG of SCI mice. (A). The expression level of phosphorylated JUN in DRG and SDH of naive, sham, and SCI mice was measured by Western blot. (B) Quantitative analysis of the expression of phosphorylated JUN in DRG and SDH in naive, sham, and SCI (sham DRG 1.0 ± 0.26, SCI DRG 1.1 ± 0.40; sham SDH 1.1 ± 0. 48, SCI SDH 1.2 ± 0.50). (C–N) Immunostaining for phosphorylated JUN (green) and DAPI (red) staining on spinal cord sections derived from naive (C–F), sham (G–J), and SCI mice (K–N). (E-F, I–J) and (M–N) are the high magnification views of the boxed area in (C,D), (G,H), and (K, L), respectively. The inset (right) in (N), which is the high magnification view of the boxed area in (N) (left), shows phosphorylated JUN in the nucleus. (O) The number of neurons expressing phosphorylated JUN in laminae I–VI of naive, sham and SCI mice. (P–U) Immunostaining for phosphorylated JUN (green) and DAPI (red) staining on DRG sections derived from naive (P, S), sham (Q, T), and SCI (R, U) mice. (S–U) are the high magnification views of the boxed area in (P–R), respectively. The inset (right) in (U), which is the high magnification view of the boxed area in (U, left) shows phosphorylated JUN in the nucleus. (V) The number of neurons expressing phosphorylated JUN in DRG of naive, sham, and SCI mice. # represents the comparison between SCI and sham. * represents the comparison between naive and SCI or sham. Data are shown as Mean ± SD, n = 3. #p < 0.05, *p < 0.05, **p < 0.01, unpaired t-test or one-way ANOVA. n.s. not significant. Scale bar = 100 μm.

PMC10020601

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5e215178dbc2412eb7e4126b29cfc1e8

Schematic view of mechanism underlying SCI-induced NP. (A). SCI induced the upregulation of NGF, and consequently increased phosphorylated JUN and the upregulation of Nav1.7 in SDH and DRG neurons of mice. (B). Nav1.7 selective blockers attenuated SCI-induced NP in mice through inhibiting the activity of Nav1.7 in both the spinal cord and DRG.

PMC10020601

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88966ea211b24defa8104d8dd58b94e0

Timeline of major retinal imaging instruments and landmark artificial intelligence applications in glaucoma. Introduction of landmark imaging instruments are listed in blue and AI events are provided in black. AI, artificial intelligence; OCT, optical coherence tomography; OCTA, OCT angiography.

PMC10020779

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f66dd2cfc769400fbd70b3669e38c4ff

Evolution of AI in glaucoma. First row: Image processing and expert systems were used to identify glaucoma landmarks or features (such as cup-to-disc ratio or hemorrhages) from retinal images with the assistance of a glaucoma specialist and glaucoma landmarks are identified. Second row: Numerical parameters like raw visual fields (VFs), intraocular pressure (IOP), and age from normal and glaucomatous subjects (presented as N and G) are input to a conventional machine learning model (e.g., neural network) without glaucoma specialist assistance and diagnosis is made. Third row: Image processing and expert systems were used to quantify glaucoma landmarks (extract features) with the assistance of a glaucoma specialist then quantified parameters (features) from normal and glaucomatous subjects are fed to a conventional machine learning model to make diagnosis. Fourth row: Retinal image is fed to an end-to-end deep learning model and the diagnosis is made without assistance from a glaucoma specialist.

PMC10020779

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4e2afd5d54c54aa88a2c6d796c07f892

Visual field (VF) and optical coherence tomography (OCT) image quantification. Top: VFs were quantified to 18 prominent patterns of VF loss based on classical archetypal analysis. Bottom: OCT circle scans were quantified to 16 patterns of RNFL loss based on deep archetypal analysis.

PMC10020779

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65226d75ec19428eaf9391585b88bac9

Geographical distribution of melioidosis cases from 2013–2018 Udupi District, Karnataka, India (n = 50).Base map republished from [20] under a CC BY license, with permission from Karnataka State Remote Sending Application Centre (KSRC), original copyright KSRC, 2022.

PMC10021467

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f6806cd9b1854cdb9fefc08e38432721

Monthly distribution of average melioidosis cases and rainfall from 2013–2018, Udupi district, Karnataka, India (n = 50).

PMC10021467

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71bcc270c7aa4e7a96d805c485814f65

PRISMA flow diagram, article selection process.

PMC10021551

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168cb2fc8e674d9ba3f4765be2bf603d

Proportion of individuals in different level of nutritional status based on the BMI indices.

PMC10021695

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12d5205c7b3248dbac94a56bf7ed0537

List of NCDs studied and their magnitude among participants attending chronic outpatient NCDs care, Bahir Dar, Ethiopia (N = 1432).

PMC10021695

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98d698ec3e9243f7ba7f7a8543c386dc

Patterns of NCDs morbidity among individuals attending chronic NCDs care in Bahir Dar, Ethiopia (N = 1432).

PMC10021695

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Number of individuals classified in different categories of health-related QoL.

PMC10021695

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2926cb5bc91146a0ab1f59f8c3a2018f

Graphic presentation of the relationship between QoL, functioning and multimorbidity.

PMC10021695

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5b98719ca8b544cea39afafe75bcf830

Localisation and diversity of natural water sites used by local population.(A) Localisation of all the unprotected and natural water sites visited in the study area. Datas for the administrative boundary were provided by Ministry of Territory Planning of Benin (https://gadm.org/download_country_v3.html). The map of rivers was provided by Ministry of Mines, Energy and Hydraulics (http://www.hydrosciences.fr/sierem/produits/gis/Oueme.asp). (B) Examples of unprotected water sources. (C) Classification of water sites by type.

PMC10021984

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d705765061a244d684ea3431e159ec8a

Identification of human activities at natural water sites.(A) Location of the 11 prospected water sites. Datas for the administrative boundary were provided by Ministry of Territory Planning of Benin (https://gadm.org/download_country_v3.html). The map of rivers was provided by Ministry of Mines, Energy and Hydraulics (http://www.hydrosciences.fr/sierem/produits/gis/Oueme.asp). (B) Illustrations of human activities related to unprotected water. (C) Frequency of activities observed at the water sites. (D) Age and sex distribution of people observed at water sites. (E) PCA of water-related activities, age and sex distribution.

PMC10021984

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b85f2ceb679748faa22a9e64675c361d

Factors involved in Buruli ulcer disease.(A) Location of Buruli ulcer patients participating in the prospective case-control study. Datas for the administrative boundary were provided by Ministry of Territory Planning of Benin (https://gadm.org/download_country_v3.html). (B) Age and sex distribution of Buruli ulcer patients. (C) Proportion of patients using clean water from drilled boreholes/wells and/or water from unprotected water sites. (D) Mapping of lowlands by spatial analysis. Datas for the altitude and land surface are available on https://earthexplorer.usgs.gov/ (NASA), and https://apps.sentinel-hub.com/eo-browser/ (European Spatial Agency). (E) Number of Buruli ulcer cases according to distance from the river or a lowland water site.

PMC10021984

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e117cbbcaf8242218e35378ef023c739

Detection of M. ulcerans at water sites.(A) Location of water sites from which environmental samples were collected. In green, water sites at which none of the pooled samples was positive for M. ulcerans DNA, and, in red, water sites at which at least one pooled sample tested positive for M. ulcerans DNA. Datas for the altitude and land surface are available on https://earthexplorer.usgs.gov/ (NASA), and https://apps.sentinel-hub.com/eo-browser/ (European Spatial Agency). Datas for the administrative boundary were provided by Ministry of Territory Planning of Benin (https://gadm.org/download_country_v3.html). (B) Proportion of positive water sites located in lowlands.

PMC10021984

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Link between the area under water and endemicity.(A) PCA analysis with monthly data for water coverage by district and endemicity. (B) Example of surface water area for two districts: Gangban and Dame-Wogon. (C) Projection of axis 1 of the PCA. (D) projection of axis 2 of the PCA. (E) Incidence of Buruli ulcer in the districts of the study area. Datas for the administrative boundary were provided by Ministry of Territory Planning of Benin (https://gadm.org/download_country_v3.html).

PMC10021984

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7f993852e92648babd0c997da3fd9e39

Census data (2013) & past vaccination tallies from Child Health Day 6-months earlier vs estimated denominators.

PMC10022106

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db86fdc21f2948129c0c599d55993d91

Frequentist estimates of polio vaccination rates by commune.

PMC10022106

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Bayesian estimates of polio vaccination rates by commune.

PMC10022106

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63fd1591b9574895abf3b7046a6889c9

Sensitivity analysis of polio vaccination rates by commune.

PMC10022106

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f50675301f5e4f71a7d4bbd829e7573c

Priming NK cells with IL-12, IL-15, and IL-18 prompts trained memory-like phenotypic alterations among different NK subpopulations. A Schema of isolating NK cells from peripheral blood mononuclear cells and priming them with IL-12, IL-15, and IL-18 or low concentrations of IL-15 for 16 h, washing and then resting in a low concentration of IL-15 to allow for differentiation in vitro. B Representative flow plots showing the frequency of CD56 positive cells before and after positive separation. C Representative flow plots showing c-NK cells and CIML NK cells. Insert values are the percent positive of indicated markers for c-NK cells and CIML NK cells that fall within the indicated CD56 and CD16 gate. D The bar graphs show a comparison of c-NK and CIML NK cells from a representative individual on 3 NK cell phenotypic markers of different NK cell subsets. All values shown are mean ± s.e.m. of triplicate or duplicate measurements and have been repeated 3 times with similar results

PMC10022190

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Preactivation with IL-12, IL-15, and IL-18 induces CD25 and CD137 up-regulation on CIML NK. A Schema of in vitro experiments. Purified NK cells were activated with IL-12, IL-15, and IL-18 or control-treated for 16 h and washed. After differentiating for 6 days, NK cell phenotype and functionality were assessed. B Representative flow plots showing the expression of CD25 and CD137 on NK cells after stimulation with the IL-12, IL-15, and IL-18 or low concentrations of IL-15 for 16 h and cocultured with tumor targets (K562, HepG2, and SK-Hep-1 cells) on day 7 performed at an effector-to-target ratio (E: T) of 2:1 for 24 h. Insert values are the percent positive of indicated markers for c-NK cells and CIML NK cells. C Graphs show the quantification of FACS analysis data. Data are presented as means ± s.e.m. Statistical significance was calculated by unpaired two-sided t-test

PMC10022190

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CIML NK cells exhibit enhanced functional responses against hepatocarcinoma targets. Representative bivariate mass cytometry plots of IFN-γ (A) and TNF-α (C) expressed by c-NK cells and CIML NK cells stimulated by HepG2 cells at the designated effector-to-target ratios (E: T). Summary of data (mean ± s.e.m.) from the same donor showing percentages of IFN-γ-positive NK cells (B) and TNF-α–positive NK cells (D) at an E: T of 2:1. E The bar graphs show a comparison of IFN-γ secretion of c-NK cells and CIML NK cells following incubation with the indicated cell lines for six hours in which K562 cells worked as a positive control group. F The cytotoxic reactivity of c-NK and CIML NK cells was measured using CFSE/PI cytotoxicity assay, the target cells were HepG2 (left) and SK-hep-1(right), respectively. Statistical significance was calculated by an unpaired two-sided t-test. TNF-α tumor necrosis factor-alpha, IFN-γ interferon-gamma

PMC10022190

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CIML NK-iRGD showed improved tumor infiltration capacity and antitumor efficiency in 3D tumor spheroids. A The constructed MCSs of iRGD-receptors-positive gastric cancer cell line HGC27. The image was obtained under 4 × magnification. The scale bar was 100 μm. B Representative morphological assessment of HGC27-MCSs was exposed to indicated CFSE stained NK cells at an effector to target cell ratio (E: T) of 5:1 calculated on the initial number of spheroids inoculated for 6 h before confocal microscopy. C Summary of data showing the infiltrated depth of HGC27-MCSs by indicated NK cells for 6 h by quantitative analysis of mean fluorescence intensity. Statistical significance was calculated by one-way analysis of variance (ANOVA). D Representative morphological assessment of HGC27-MCSs was destroyed by indicated NK cells at an E: T of 5:1 calculated on the initial number of spheroids inoculated for 6 h before Viability/Cytotoxicity Assay Kit staining and imaged by confocal microscopy. Viable HGC27 cells are stained with Calcein AM (green). Dead HGC27 cells are stained with ethidium homodimer III (EthD-III; red). E Summary of data from D showing Live/dead cell quantification in MCSs. Data are presented as means ± s.e.m. Statistical significance was calculated by unpaired two-sided t-test

PMC10022190

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iRGD-modified P-CIML NK cells effectively controlled tumor growth. A Experimental scheme of the antitumor experiment in vivo. Five-week-old female Babl/c-nude mice were subcutaneously injected with 5 × 106 HepG2 cells to initiate an antitumor experiment in vivo. After two weeks, 2 × 107 P-CIML NK (PBMCs were primed with IL-12, IL-15, and IL-18) cells or P–c-NK (PBMCs primed with low concentration IL-15) modified with iRGD or not were injected into the mice. This was followed by IL-2 treatment every other day for approximately 10 times, with 4 or 5 mice in each treatment group. The tumor burden of the mice was monitored. B Histograms showing the expression of αvβ3, αvβ5 and NRP-1 on HepG2 cells. C Imaging of mice with subcutaneous HepG2 tumors was conducted in vivo at various intervals post intravenous injection of CIML NK (sorted CD56 + cells) cells, which had been modified with or without iRGD. Magenta dashed lines indicate tumors. CIML NK cells labeled with DiR. D The percentage of human CD56 + cells in the tumor tissue of CIML NK-iRGD or control CIML NK-treated mice detected by flow cytometry at 24h after intravenous injection of NK cells. E Summary of data from D showing CD56 + NK cells among total cells. F The tumor volume of the mice treated with P–c-NK or P-CIML NK modified with or without iRGD. G Photos of tumors harvested from mice in all groups on day 28 after tumor inoculation. The average tumor volume (H) and tumor tissue weight (I) in different groups at the endpoint of the animal experiment. J The average weight of different groups for 28 days. Statistical significance was calculated by one-way analysis of variance (ANOVA) represented as mean ± s.e.m. NRP-1 Neuropilin-1, s.c. subcutaneous injection, i.v. intravenous injection, i.p. intraperitoneal

PMC10022190

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Design of this study

PMC10022191

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Mean values of the plasma concentrations of Canocapavir over time in each treatment cohort (mean ± standard deviation)

PMC10022191

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Changes of HBV virologic markers from baseline during treatment with Canocapavir or placebo. A–C Hepatitis B virus (HBV) DNA, D–F HBV pregenomic RNA (pgRNA), G hepatitis B surface antigen (HBsAg), H hepatitis e surface antigen (HBeAg), I hepatitis B core-related antigen (HBcrAg)

PMC10022191

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b1200a89024d40908ea28a60b5b9fe30

Conceptualization model: The decision process for each risk management method chosen.

PMC10022342

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Contraceptive indicators between 1998 and 2016 (SADHS, 2016).

PMC10022780

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af24987077054df39a2bfdde18eea7a5

Pretreatment. A, Preoperative images for a 60-year-old man with cystic lymphatic malformation in the right thigh area. Four small yellow triangles indicate the area where the cyst existed. The white line shows the plane of MRI view. B, MRI on T2-weighted images showed a well-described extraarticular multi lobular cystic malformation above the knee region with a maximum diameter of 16 cm, located on the anterior side of the right femoral bone under the muscle layer, without cortical bone destruction and adhesions to surrounding tissues.

PMC10022848

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Posttreatment. A, Lymphedema was improved with decreasing volume of lower limbs. The white line shows the plane of MRI view. B, MRI on T2-weighted images showing that the cystic lymphatic malformation resolved with no recurrence during 2 years of follow-up.

PMC10022848

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094eb2bfee0440eb82fbb8a020284d21

Effect of high- vs. low-dose IV iron on fatal and non-fatal MI (time-to-first event analysis). (A) Fatal and non-fatal MI; (B) type 1 MI; (C) type 2 MI; (D) NSTEMI; and (E) STEMI.

PMC10022850

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Effect of high- vs. low-dose IV iron on fatal and non-fatal MI (recurrent event analysis, i.e. first and subsequent events).

PMC10022850

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b447048894c4405aa6551a9f8cc0e01f

Comparison of the three platforms. (a) Schematic diagram of TIIA. (b) Schematic diagram of LFIA. (c) Schematic diagram of HCM. NC stands for negative control.

PMC10023210

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(a) The centrifugal tray was modified by the commercial pulling-force spinning top. (b–c) Experimental results of before (b) and after centrifugation (c). The concentration of N protein was 100 ng/mL. (d) The centrifugal speeds over time were performed by ten independent individuals. (e) Statistical analysis of velocity in plateau period of ten independent individuals.

PMC10023210

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Optimization of condition in the homogeneous immunoreaction and centrifugal process. (a) The result of immunoreaction under different red latex nanobeads concentration (a), incubation time (b), centrifugation time (c), and centrifugation speed (d).

PMC10023210

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The sensitivity testing of two systems. (a) The photographic images of the LFIA after loading different concentrations of N protein of SARS-CoV-2. (b) The statistical result of (a), n = 3. (c) The photographic images of the HCM after loading different concentrations of N protein of SARS-CoV-2. (d) The statistical result from HCM of different concentrations of N protein of SARS-CoV-2, n = 3.

PMC10023210

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(a–b) The photographic images and corresponding statistical results (b) of the HCM after loading different samples (n = 3). (c) The results of the HCM for clinical samples. CS1-CS4 indicates clinical negative samples and CS4-CS9 indicates positive clinical samples. Each sample was tested three times. The inset represents a schematic diagram of clinical sample testing on HCM. (d) The statistical results of (c). NS and PS indicate negative and positive samples, respectively.

PMC10023210

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286e3044bec444f2b49420e7dd5ec41a

Design and characterization of the orthogonal transcription system.a The binding of the orthogonal RNAP to its cognate promoters (with binding affinity KA) is a limiting step for protein synthesis in all organisms. The capping enzyme fused to RNAP ensures nuclear export and translation efficiency of the synthetic transcripts in mammalian cells. b The reporter gene expressions for the T7 promoter with RNAP-null, RNAP-only, RNAP-dead capping enzyme and RNAP-active capping enzyme. c Scheme of the quantitative measurement circuit composed of a monomeric RNA polymerase (RNAP) fused to a capping enzyme driven by the EF1α promoter and a set of T7 promoter variants in two separated plasmids. d Linearity between expression measured by flow cytometry and expression predicted by model in CHO cells transfected with 0.7 μg T7 RNAP plasmid. e Linearity between expression measured by flow cytometry and expression predicted by model in CHO cells transfected with 0.07 μg T7 RNAP plasmid. Data represent the mean ± SD (n = 3) f Comparison of expression from six different orthogonal promoters driven by cognate RNAPs and three widely-used mammalian promoters (CMV, EF1α and SV40) driven by the endogenous RNAP. Data represent the mean ± SD (n ≥ 6). g Characterization of a 6 × 6 orthogonality matrix of host-independent RNAPs and their cognate promoters. All data represent the mean ± SD (n ≥ 3). Source data are provided as a Source data file.

PMC10023750

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659c0b70e9324b14b2b8e3c9d4d622ff

Resource competition model in the two-reporter system.a The genetic circuit design for the RNAP-driven expression of two reporter genes. b Characterization of resource competition by two co-expressed reporters in HEK293T. Experimental data and model predictions are compared at varying activities for the two reporters. Data represent mean values of more than three biological replicates. c Schematic of different promoters competing for the same pool of free RNAP ([RNAP]free). d Heatmap showing the concentration of [RNAP]free as calculated by the biochemical model with resource competition. Promoter activities are decreased as shown by the triangles for two reporter genes. Source data are provided as a Source data file.

PMC10023750

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ddaaa26c7ef0488b82a30a5df48cf027

Gene-order effect for three-gene expression and predictions of the resource-competition model.a Schematic of the three-reporter characterization system with all possible orders of the transcriptional units. b Relative activity of blue, yellow and red fluorescent proteins of each reporter gene (bfp, yfp, and mCherry) based on their respective expression level in construct #1. Data represent the mean ± SD (n = 4). c Schematic of the circuit for the performance evaluation of three-reporter gene expression system. d Three-dimensional graph of the combinatorial promoter library with different binding affinities to the cognate RNAP. Different colors show different z-axis (Pbfp value). e Correlation between predicted promoter activities and experimental fluorescent intensities on a log–log scale for the three reporter genes. Data represent the mean ± SD (n = 3). Source data are provided as a Source data file.

PMC10023750

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Stoichiometry optimization of the three components for the influenza A subtype H1N1 VLP via the predictive model.a Schematic of the recombinant VLP in which NA and M1 proteins are fused with mCherry and GFP reporter proteins, respectively. b Transmission electron microscopy images of influenza VLPs. Scale bar, 50 nm. c Representative images of the VLPs produced from the three-gene system. Fluorescent microscopy was used to observe co-localization events of NA and M1. Scale bar, 10 μm. d Schematic of the plasmid constructions for VLP production. e Relative VLP yields for 21 selected promoter-gene combinations and 3 widely-used mammalian promoters (EF1α, CMV and SV40). The reference VLP yield was of the combination of the strongest promoters and was assigned as 100%. Data represent the mean ± SD (n = 3). f Predicted VLP yields in all 157,464 combinations of the available HA, M1 and NA promoters. The reference combination is normalized to 1. The dashed circle marks the rough boundary of the promoter combinations with yields higher than that of the reference. Stars indicate the combinations selected for experimental validation. g Comparison of experimental VLP yields with model predictions. Data represent the mean ± SD (n = 3). Source data are provided as a Source data file.

PMC10023750

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9636e337bff046d78d87238f0a8b0b23

Identifying stromal Tumor Infiltrating Lymphocyte (TIL) regions in a H&E-stained WSI of breast cancer. (A) H&E-stained WSI of breast cancer. (B) Example of a region of tissue. (C) Example of stromal TILs (green solid arrow) and cells that are no TILs (red dotted arrow). The image shows how it can be challenging to visually differentiate TILs from other cells. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

PMC10025040

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Workflow of the approach developed in the study. Starting from H&E images from HER2+ and TN breast cancer, the approach starts with the segmentation of nuclei within tumor-associated associated stroma (A). Pathomic features were then extracted from cell segmentation within tumor-associated stroma (B). After the feature selection step, five classification models were trained (C) and their performance evaluated (D).

PMC10025040

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d584981191fa46c999a7b00b82372b22

Selected pathomic features (n = 21) after the three steps of feature selection. Surviving features were reported in bold. Abbreviations: FSD = Fourier Shape Descriptor; MAD = Median Absolute Deviation; IDM = Inverse Difference Moment; IMC = Information Measure of Correlation.

PMC10025040

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Receiver Operating Characteristic (ROC) curves of prediction models without resampling (A), with SMOTE-resampling (B) and with downsampling (C). Abbreviations: LDA = Linear Discriminant Analysis; KNN = K-Nearest Neighbour; DT = Decision Tree; RF = Random Forest; MLP = Multi-layer Perceptron.

PMC10025040

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Comparison of AUCROCs between different classification models and different resampling techniques. Abbreviations: SMOTE = synthetic minority oversampling technique; LDA = Linear Discriminant Analysis; KNN = K-Nearest Neighbour; DT = Decision Tree; RF = Random Forest; MLP = Multi-layer Perceptron.

PMC10025040

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a Antoine Lacassagne Cancer Centre. Niza. b Medanta applicator AEOLO. c Pamplona applicator d Benidorm template

PMC10025210

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3D printed applicator. Courtesy of Dr. Elena Villafranca

PMC10025210

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Use of scout view to determine the needle tip on CT images

PMC10025210

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MRI images showing a “ballooning” artifact at the end of the titanium needle

PMC10025210

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Reconstruction method for titanium needles based on MRI images with the aid of MRI markers (A-vitamin pellets)

PMC10025210

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Example showing differences between geometrical and point-based optimisation

PMC10025210

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Schematic representation of the investigated training set designs for the prediction of location 1.

PMC10025381

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Correlation between observed and predicted yield (A) and moisture (B) for training set designs CV3(within environment with no previous data) and CV6(within year with no previous data) in years 2013 to 2015 at locations 1 to 7. The median of 100 simulations for the prediction method RR-BLUP is shown on top of the respective boxplots.

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Correlation between observed and predicted yield (A) and moisture (B) for training set designs CV3(within environment with no previous data) and CV6(within year with no previous data) for one year, and training set designs CV1to CV8across one or two years. The median of 100 simulations for the prediction method RR-BLUP is shown on top of the respective boxplots.

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Growth characteristics of longan fruits and their quantitative statistical scheme.

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Solution for longan fruit tree yield statistics.

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Schematic diagram of the image acquisition mode.

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Examples of the UAV images.

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Diagram of the whole process.

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Flow chart of Cluster_Fruits and Single_Fruits quantity prediction. (A) The flow chart of Single_Fruit quantity statistics in a Cluster_Fruit, (B) The flow chart of Cluster_Fruit quantity statistics in a single fruit tree.

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Clustering results of the target bounding boxes in the two datasets.

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P-R curves and F1 scores of different detection methods.

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Cluster_Fruit detection results of the CF-YD model under different scenes.

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Single_Fruit detection results of the SF-YD model under different scenes.

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Cluster_Fruit counting results of the CF-YD model under different scenes.

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Single_Fruit counting results of the SF-YD model under different scenes.

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The fitting results of the actual and identified numbers of fruits.

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Statistical information of Cluster_Fruits and Single_Fruits. (A) The actual numbers, identified numbers and predicted numbers of Cluster_Fruits on six longan trees, (B) The actual numbers, identified numbers and predicted numbers of Single_Fruits on six Cluster_Fruits, (C) Error rate information of Cluster_Fruit on six trees, (D) Error rate information of Single_Fruit on six Cluster_Fruits.

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Liposarcoma (a) Gross photograph showing a greyish yellow tumor in a mastectomy specimen (marked with arrow), (b)Histopathology shows a tumour composed of mature adipocytes of variable size along with highly pleomorphic cells with vesicular nuclei, prominent nucleoli and scant cytoplasm. There are lip blasts with few bizarre and multinucleated forms were also seen, (c) Tumour cells showed marked pleomorphism with spindle to oval nuclei and brisk mitosis (d) extensive areas of necrosis. (H and E, b:100X c&d: 400x)

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Tumor cells are positive for (a) Vimentin, and (b) focally for S-100 (×400)

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Magnetic resonance imaging (MRI) showing lobulated mass lesion in the sellar-suprasellar region, Inset - MRI contrast enhancement of the mass lesion

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Squash cytology showing inflammatory infiltrate with few entangled epithelial cells (H and E, ×10)

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Pituitary glandular epithelium with infiltration by lymphocytes, few plasma cells, and macrophages (H and E, ×10) Inset - Higher power of the same (H and E, ×40)

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