Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Outline of study time-course and sample processing for EV isolation. Antemortem plasma samples with postmortem pathological confirmation of neurological diagnoses were processed to isolate cell-specific EVs using our mTENPO microfluidic platform, alongside plasma protein biomarkers using commercial digital ELISA, for patients with LBD ( n = 30), AD ( n = 31), AD/LBD ( n = 30), AD/ALB ( n = 19), and controls ( n = 27). B The mTENPO platform, illustrating the external magnet, inlet reservoir, outlet ports, and tubing connections to syringe pumps. Syringes are connected to the waste outlet for blocking, washing, and sample addition steps, and then replaced and switched to the lysate outlet before captured EVs are lysed on-chip. The inset shows a photo of the mTENPO chip with a quarter for scale. C Schematic of operation of the mTENPO platform for cell-specific EV isolation using antibody-labeled magnetic nanoparticles (MNPs) for GluR2+ (top) and GLAST+ (bottom) EV pulldowns. D Scanning electron microscopy (SEM) images of GluR2+ EVs immobilized on the edges of pores of the mTENPO device’s surface. E SEM images of GLAST+ EVs immobilized on the edges of pores of the mTENPO device’s surface. F Representative cropped western blot images showing protein expression of GluR2, GLAST, and EV-associated marker TSG101 using mTENPO-isolated GluR2+ or GLAST + EV lysates from n = 2 human plasma samples. Full-length western blot images are shown in Supplementary Fig. .

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Isolation, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Blocking Assay, Labeling, Electron Microscopy, Western Blot, Expressing, Marker

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for biomarkers with Benjamini-Hochberg FDR-corrected P value < 0.1. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending fold-change. B Volcano plot demonstrating differential expression of GluR2+ EV miRNAs, GLAST + EV miRNAs, and plasma proteins. C Venn diagram showing overlap in FDR P value significant miRNAs ( P value < 0.1) between GluR2+ EVs and GLAST+ EVs. D Top 30 biomarkers in all compartments ranked by descending area under the curve (AUC). Error bars represent standard error from bootstrapping 10x.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Quantitative Proteomics, Clinical Proteomics

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: GO and KEGG pathway analyses were performed on differentially expressed miRNAs using DIANA miRPath v4.0 using the TarBase v8.0 database. FDR P values for identified GO terms and KEGG pathways were calculated using a one-sided Fisher’s exact test and considered significant at P value < 0.05. The top 10 (ranked by number of target genes) terms within each of the three GO categories (BP, CC, MF) and top 10 (ranked by number of target genes) KEGG pathways were identified for each pulldown. A Top 10 terms within each GO category for GluR2+ EV miRNAs. B Top 10 KEGG pathways for GluR2+ EV miRNAs. C Top 10 terms within each GO category for GLAST + EV miRNAs. D Top 10 KEGG pathways for GLAST + EV miRNAs. In all panels, each bar is labeled to the right with the number of differentially expressed miRNAs associated with the given GO term or KEGG pathway.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Labeling

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for LASSO-selected biomarkers. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending AUC. B Kendall correlation staircase plots identifying the extent to which biomarker information was correlated between the LASSO-selected GluR2+ EV, GLAST + EV, and protein biomarkers. Biomarkers are sorted within compartments by AUC. The inset shows the correlation distribution of Kendall’s τ, where the dotted line represents the median count. C LASSO panel accuracy versus panel size for classifying LBD versus AD, shown in blue; accuracy is assessed through tenfold cross-validation, with error bars representing standard error from 5 repeats of panel training on the LBD vs AD patient groups. Average accuracy and standard error for control experiments performed by scrambling patient labels 10x are shown in orange. D LASSO panel AUC versus panel size for classifying LBD versus AD, shown in blue with error bars as described in ( C ). Average AUC and standard error for the same control experiments described in ( C ) are shown in orange. E AUCs for the 15-marker LASSO panel and individual LASSO biomarkers, sorted by descending AUC. Error bars represent 95% confidence intervals, calculated from 5x repeats of tenfold cross-validation for the 15-marker panel or from bootstrapping 10x for individual markers.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Biomarker Discovery, Control, Marker

Journal: 3 Biotech

Article Title: Gandouling protects against hepatic fibrosis in Wilson disease through the lncRNA-SNHG7/miR-29b/DNMT3A pathway

doi: 10.1007/s13205-026-04769-0

Figure Lengend Snippet: GDL inhibits autophagic activity in LX-2 cells. A Western blotting analysis of Beclin-1, LC3-II/LC3-I, and p62 expression ( n = 3). GAPDH was used as a loading control. Data are presented as means ± SD. ## p < 0.01 vs. control group; * p < 0.05, ** p < 0.01 vs. CuSO 4 group. B Representative fluorescent images of cells following mCherry-GFP-LC3 adenovirus infection (Scale bar: 10 μm). In the merged image, yellow spots indicate autophagosomes, while red spots indicate autophagic lysosomes. The degree of autophagic flux can be seen by the number of different color spots

Article Snippet: The membranes were then incubated overnight with antibodies against DNMT3A (Bioss, Item No. bs-23029R, 1:2000, Rabbit, 1:1000, Rabbit), p-DNMT3A (Bioss, Item No. bs-14399R, 1:2000, Rabbit), α-SMA (Bioss, Item No. Bsm-33187 M, 1:2000, Mouse; No. bs-0189R, 1:1000, Rabbit), Collagen I (Bioss, Item No. bs-7158R, 1:1000, Rabbit; No. AB260043 , 1:1000, Rabbit), Beclin-1 (Abcam, Item No. ab210498, 1:2000, Rabbit; No. ab62472, 1:1000, Rabbit), LC3B (CST, Item No. 3868s, 1:1000, Rabbit; No. 43566 S, 1:1000, Rabbit), and p62 (Bioss, Item No. bs-2951R, 1:1000, Rabbit) at 4°C with gentle shaking.

Techniques: Activity Assay, Western Blot, Expressing, Control, Infection

Journal: International Journal of Molecular Sciences

Article Title: Hydrogen Mitigated Doxorubicin-Induced Liver Injury via Nrf2/HO-1 Pathway Activation

doi: 10.3390/ijms27062774

Figure Lengend Snippet: Hydrogen activated the Nrf2/HO-1 pathway to attenuate liver injury in DOX mice. ( A , B ) Nfe2l2 and Hmox1 gene expression in liver tissue (n = 3). ( C – E ) IHC staining and statistics of Nrf2 and HO-1 protein (scale bar = 100 µm, n = 3). ( F – I ) Expression and statistics of Keap1, Nrf2, and HO-1 protein levels measured by Western blot (n = 4). The results are presented as the mean ± SEM. * p < 0.05 vs. Con group. # p < 0.05 vs. DOX group.

Article Snippet: Antibodies: Bcl-2 ( GB153375 , Servicebio, Wuhan, China), Bax (GB11007, Servicebio, Wuhan, China), Caspase 3 (#14220, Cell Signaling Technology, Danvers, MA, USA), MDA (ab243066, Abcam, Cincinnati, OH, USA), 4-HNE ( ARG23717 , Arigo Biolaboratories, Hsinchu, Taiwan), IL-6 (DF6087, Affinity, Cincinnati, OH, USA), NLRP3 (BA3677, Boster, Wuhan, China), Nrf2 ( GB115673 , Servicebio, Wuhan, China), HO-1 (GB12104, Servicebio, Wuhan, China), Keap1 (bs-4900R, BIOSS, Shanghai, China), and α-Tubulin (GB11200, Servicebio, Wuhan, China).

Techniques: Gene Expression, Immunohistochemistry, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Hydrogen Mitigated Doxorubicin-Induced Liver Injury via Nrf2/HO-1 Pathway Activation

doi: 10.3390/ijms27062774

Figure Lengend Snippet: The schematic diagram of hydrogen protection against DOX-induced liver injury. DOX has been observed to provoke biochemical alterations and pathological abnormalities in the liver. Specifically, DOX facilitates the generation of ROS and promotes mitochondria-dependent cell apoptosis. Additionally, DOX impedes the reduction in the expression of Keap1 and Nrf2, thereby inhibiting the downstream antioxidant signaling pathways of Nrf2, including HO-1, CAT, and T-SOD. Concurrently, it enhances the expression of lipid peroxidation products such as MDA and 4-HNE. Furthermore, DOX instigates inflammatory processes and augments the release of pro-inflammatory cytokines. In contrast, hydrogen exerts a protective effect on DOX-induced liver dysfunction by modulating Nrf2, which mitigates oxidative stress and inflammatory responses.

Article Snippet: Antibodies: Bcl-2 ( GB153375 , Servicebio, Wuhan, China), Bax (GB11007, Servicebio, Wuhan, China), Caspase 3 (#14220, Cell Signaling Technology, Danvers, MA, USA), MDA (ab243066, Abcam, Cincinnati, OH, USA), 4-HNE ( ARG23717 , Arigo Biolaboratories, Hsinchu, Taiwan), IL-6 (DF6087, Affinity, Cincinnati, OH, USA), NLRP3 (BA3677, Boster, Wuhan, China), Nrf2 ( GB115673 , Servicebio, Wuhan, China), HO-1 (GB12104, Servicebio, Wuhan, China), Keap1 (bs-4900R, BIOSS, Shanghai, China), and α-Tubulin (GB11200, Servicebio, Wuhan, China).

Techniques: Expressing, Protein-Protein interactions