Journal: Journal of Advanced Research

Article Title: An injectable nano-hydroxyapatite-incorporated hydrogel with sustained release of Notoginsenoside R1 enhances bone regeneration by promoting angiogenesis through Notch1/Akt signaling

doi: 10.1016/j.jare.2025.05.025

Figure Lengend Snippet: (A) The angiogenic-related gene expression quantified by qRT-PCR at day 4, day 7 and day 10. (B) The angiogenic proteins of VEGF and VEGFR-2 analyzed by western blot. (C)semiquantitative analysis of VEGF and VEGFR-2 protein expression. (D) The critical markers Notch1, Hes1 involved in the Notch 1 pathway analyzed by western blot and their semiquantitative analysis (E).(F)Western blot and semiquantitative analysis (G) of Akt protein expression.(H)The gene expression of Akt and VEGF under NGR1 stimulation with Notch 1 inhibition. (I) The gene expression of Notch 1 and VEGF under NGR1 stimulation with Akt inhibition. (J) Western blot and semiquantitative analysis (K) of Akt protein expression induced by NGR1 under Notch 1 inhibition. *p < 0.05, **p < 0.01, ***p < 0.001, ns, not significant.

Article Snippet: Subsequently, Notch 1 inhibitor (HY-1302, MCE, USA) and Akt inhibitor (HY-10358, MCE, USA) were applied to evaluate their effects on angiogenic-related gene expression.

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, Expressing, Inhibition

Journal: bioRxiv

Article Title: C-terminus CD28 phosphorylation (Y218) modulates IL-2 secretion and antitumor effect of CAR-T cells

doi: 10.64898/2026.01.28.701378

Figure Lengend Snippet: A . Top panel: Schematic representation of the protocol used to inhibit ITK activity. PSCA CAR-T cells were pre-treated for 2 hours with ITK inhibitors 1μM (BMS-509744, Ibrutinib or GNE-9822) or DMSO as control and then cocultured with HPAC WT cells for 0 or 10 minutes. Left panel: Representative Western blot of CD28 pY218 in CAR-T cells after stimulation with HPAC WT cells. Right panel: Normalized CD28 pY218 intensity with respect to CAR (CD3ζ). Significance was determined using one-way ANOVA. * = P<0.05. Each symbol represents an independent experiment from 3 different healthy donors. Data is represented as the mean ± standard deviation (SD). B. Top panel. Schematic representation of the experimental protocol. ITK expression was disrupted using CRISPR/Cas in Jurkat cells. WT and ITK-KO Jurkat cells were transduced to express a PSCA-specific CAR and cocultured with HPAC WT cells for 0 or 10 min. Left panel. Representative membrane of CD28 pY218 in CAR-T Jurkat cells after stimulation. Right panel. Normalized CD28 pY218 intensity quantified by densitometry (representative plot) and CD28 pY218 at 10 min post-stimulation. Significance was determined by paired t-test. ** = P<0.01. Each symbol represents one of 3 independent experiments, performed with different donor T cells.

Article Snippet: BMS-509744 and Ibrutinib were purchased from MedChemExpress (HY-11092 and HY-10997 respectively) and GNE-9822 was provided by Genentech under an MTA.

Techniques: Activity Assay, Control, Western Blot, Standard Deviation, Expressing, CRISPR, Membrane

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Fatty acid metabolism is significantly disrupted in cerebrospinal fluid (CSF) of SAH patients, with fatty acid‐binding protein (A‐FABP) enrichment. (A) Schematic workflow of targeted fatty acid metabolomics analysis based on human CSF samples (subarachnoid haemorrhage [SAH] group, n = 40; control group, n = 30). (B and C) The orthogonal partial least squares discrimination analysis (OPLS‐DA) reveals clear separation between the SAH (blue) and control (red) groups. (D) Heatmap of the differentially expressed fatty acid metabolites between the two groups. Rows represent individual fatty acids; colour intensity reflects relative abundance (red: high; blue: low). (E) The volcano plot presents significantly altered fatty acids, including C20:4n6 (arachidonic acid [AA]), C18:2n6c (linoleic acid [LA]) and C16:0 (palmitic acid [PA]). (F) Kyoto Encyclopaedia of Genes and Genomes ( https://www.kegg.jp/kegg/pathway.html ) (KEGG) pathway enrichment analysis of dysregulated fatty acid metabolism. (G) The regulation of lipolysis in adipocytes in KEGG pathway database (map04923), with A‐FABP highlighted in red. Source : Schematic workflow created with BioRender.com.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Binding Assay, Control

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Elevated levels of fatty acid‐binding protein (A‐FABP) in cerebrospinal fluid (CSF) predict subarachnoid haemorrhage (SAH) severity and poor outcomes. (A) Schematic of cohort grouping (SAH: n = 40; control: n = 30). (B–D) Violin plots present levels of significantly altered fatty acids in CSF: (B) arachidonic acid (AA, C20:4n6), (C) linoleic acid (LA, C18:2n6c) and (D) palmitic acid (PA, C16:0). (E) Schematic of cohort grouping by SAH severity: (Hunt and Hess [H–H] group 1–2 level, n = 25; H–H group 3–5 level, n = 15). (F–H) Levels of (F) AA, (G) LA and (H) PA between severity low‐grade (H–H 1–2) and high‐grade (H–H 3–5) subgroups. (I–K) Correlation between significantly altered fatty acids and A‐FABP levels in CSF of SAH patients [(I) AA, (J) LA and (K) PA, n = 40]. (L) CSF levels of A‐FABP between severity low‐grade (H–H 1–2) and high‐grade (H–H 3–5) subgroups ( n = 48). (M and N) The relationship between CSF levels of A‐FABP and functional outcomes (modified Rankin Scale [mRS]) in SAH patients ( n = 48). Data are presented as means ± SD. * p < .05, ** p < .01, *** p < .001. Source : Schematic diagrams created with BioRender.com.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Binding Assay, Control, Functional Assay, Modification

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Cerebral fatty acid‐binding protein (A‐FABP) is increased in mice and predominantly colocalizes with microglia. (A and B) Representative photographs from the bottom of mice brains and H&E staining from sham and subarachnoid haemorrhage (SAH) groups. (C) Quantification of FABPs family mRNA level in the mice cortex 24 h after SAH ( n = 3). (D) Representative images of immunohistochemical staining of A‐FABP (scale bar = 100 µm, n = 6). (E) Quantification of A‐FABP mRNA level in the mice cortex after SAH ( n = 6). (F and G) Western blot images and quantitative analysis of A‐FABP expressions in the mice cortex at 3, 12, 24, 48 and 72 h post‐SAH ( n = 6). (H–L) Representative microphotographs and quantitative analyses of immunofluorescence co‐staining for A‐FABP (red) with microglia (IBA1, green), astrocyte cells (GFAP, green), neuron cells (NeuN, green) and endothelial cells (CD31, green). Scale bar = 100 µm, n = 6. Data are presented as means ± SD. * p < .05, ** p < .01, *** p < .001.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Binding Assay, Staining, Immunohistochemical staining, Western Blot, Immunofluorescence

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Genetic ablation of fatty acid‐binding protein (A‐FABP) ameliorates brain injury in mice with subarachnoid haemorrhage (SAH). (A and B) Rotarod test: (A) latency to fall on the accelerating rotarod, (B) speed at fall on the accelerating rotarod ( n = 8). (C and D) Open field test: (C) distance travelled and (D) fast movement proportion ( n = 8). (E) Percentage of brain water content ( n = 6). (F and G) Representative photographs and quantitative analysis of mice brain stained with Evans Blue ( n = 6). (H and I) Representative images and quantitative analysis of TUNEL staining with co‐staining with NeuN (red) and TUNEL‐positive neurons (green, scale bar = 100 µm, n = 6). (J and L) Representative western blot images and quantitative analyses of (K) cleaved Caspase‐3/Caspase‐3 and (L) Bcl‐2/Bax ( n = 6). Data are presented as means ± SD. * p < .05, ** p < .01, *** p < .001, **** p < .0001.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Binding Assay, Staining, TUNEL Assay, Western Blot

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Fatty acid‐binding protein (A‐FABP) exacerbates microglia‐mediated neuroinflammation in mice with subarachnoid haemorrhage (SAH). (A) The mRNA levels of pro‐inflammatory cytokines TNF‐α, IL‐1β and IL‐6 are measured in different groups ( n = 6). (B–E) Representative western blot images and quantitative analyses of (C) TNF‐α, (D) IL‐1β and (E) IL‐6 in different groups ( n = 6). (F) The mRNA levels of pro‐inflammatory cytokines TNF‐α, IL‐1β and IL‐6 in WT and knockout (KO) mice after SAH ( n = 6). (G–J) Representative western blot images and quantitative analyses of pro‐inflammatory cytokines (H) TNF‐α, (I) IL‐1β and (J) IL‐6 in WT and KO mice after SAH ( n = 6). (K) Representative microphotographs of immunofluorescence co‐staining for pro‐inflammatory cytokines (red) with microglia (IBA1, green) in WT and KO mice after SAH (scale bar = 100 µm, n = 6). Data are presented as means ± SD. * p < .05, ** p < .01, *** p < .001.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Binding Assay, Western Blot, Knock-Out, Immunofluorescence, Staining

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Inhibition of fatty acid‐binding protein (A‐FABP) in microglia attenuates neuronal apoptosis and blood–brain barrier disruption. (A and B) Representative western blot image and quantitative analysis of A‐FABP expression at 0, 12 and 24 h post‐PA (200 µM) treatment ( n = 6). (C and D) Representative western blot image and quantitative analysis of A‐FABP expression at 0, 12 and 24 h post‐AA (100 µM) treatment ( n = 6). (E) Schematic diagram of microglial culture supernatant transfer model (PA, 200 µM; BMS, 20 µM). (F and G) Representative flow cytometry images of primary neuron cells apoptosis in PA‐induced microglial culture supernatant transfer model ( n = 6). (H) Schematic representation of bEnd.3 endothelial cell co‐culture system with BV2 microglial conditioned medium (CM) treated with PA (200 µM), with or without BMS (20 µM). (I and J) Representative microphotographs of immunofluorescence and quantitative analysis in bEnd.3 cells co‐cultured with the CM of BV2 cells (scale bar = 100 µm, n = 6). Data are presented as means ± SD. * p < .05, ** p < .01, **** p < .0001.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Inhibition, Binding Assay, Disruption, Western Blot, Expressing, Flow Cytometry, Co-Culture Assay, Immunofluorescence, Cell Culture

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Fatty acid‐binding protein (A‐FABP) suppression promotes fatty acid β‐oxidation reprogramming in microglia. (A) Schematic illustration of experimental design. Control group: PBS treatment. FFA group: 200 µM palmitic acid (PA). BMS group: Co‐treatment with PA (200 µM) and BMS (20 µM). (B–D) Oxygen consumption rate (OCR) was determined in BV2 cells treated with PA, with or without BMS ( n = 6). Quantitative analyses of (C) basal respiration and (D) maximal respiration. (E–H) Glycolytic proton efflux rate (GlycoPER) was determined in BV2 cells treated with PA, with or without BMS ( n = 6). Quantitative analyses of (F) basal glycolysis, (G) basal proton efflux rate and (H) compensatory glycolysis. (I–K) Representative western blot images and quantitative analyses of (J) CPT1A and (K) ACADL in different groups ( n = 6). (L) ATP content assay in different groups ( n = 6). Data are presented as means ± SD. * p < .05, **** p < .0001. glycoPER, glycolytic proton efflux rate. Source : Schematic diagram created with BioRender.com.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Binding Assay, Control, Western Blot

Journal: Clinical and Translational Medicine

Article Title: Adipocyte fatty acid‐binding protein as a cerebrospinal fluid–accessible biomarker and druggable target in subarachnoid haemorrhage: Linking fatty acid dysregulation to microglial neuroinflammation

doi: 10.1002/ctm2.70607

Figure Lengend Snippet: Fatty acid‐binding protein (A‐FABP) mediates neuroinflammation via the JAK2/STAT3 signalling pathway. (A) Workflow diagram. Proteomic analysis of brain tissue around the subarachnoid space from the subarachnoid haemorrhage (SAH)‐treated Veh group and the SAH‐treated BMS group ( n = 4). (B) KEGG enrichment highlights the JAK‐STAT signalling pathway. (C) Volcano plot demonstrates the results of 85 differentially expressed proteins. STAT3 is labelled in green. (D–H) Representative western blot images and quantitative analyses of (E) p‐STAT3/STAT3, (F) p‐JAK1/JAK1, (G) p‐JAK2/JAK2 and (H) p‐JAK3/JAK3 in WT and knockout (KO) mice after SAH. (I–L) Representative western blot images and quantitative analyses of (J) TNF‐α, (K) IL‐1β and (L) IL‐6 in different groups. (M and N) Representative images and quantitative analysis of TUNEL staining with co‐staining with NeuN (red) and TUNEL‐positive neurons (green, scale bar = 100 µm, n = 6). Data are presented as means ± SD. * p < .05, ** p < .01, *** p < .001. C‐A1, coumermycin A1; ns, no significance. Source : Schematic diagram created with BioRender.com.

Article Snippet: Mice were gavaged orally with a selective A‐FABP inhibitor (BMS309403, #HY‐101903, MCE, USA; 15 mg/kg/day; dissolved in 4% Tween 80 and phosphate‐buffered saline) or vehicle (without BMS309403) one and 12 h after the sham or SAH surgery and then once per day for the next six consecutive days (concentration as previously described).

Techniques: Binding Assay, Western Blot, Knock-Out, TUNEL Assay, Staining