Journal: CNS Neuroscience & Therapeutics

Article Title: Sex‐Specific Effects of Dexmedetomidine on Brain Microvascular Endothelial Cell Function After Traumatic Brain Injury in Juvenile Mice

doi: 10.1002/cns.71204

Figure Lengend Snippet: Dysregulation of BMECs and the effects of DEX on body weight and sensorimotor functions. (A) Representative images from male and female Sham and TBI groups illustrate increased co‐localization of lipid peroxides (oxidative stress marker; green) with mitochondria (magenta) in isolated BMECs of both male and female TBI mice compared with Sham controls. Nuclear staining was performed using DAPI (4′,6‐diamidino‐2‐phenylindole; blue). Scale bar: 50 μm. (B) Representative images of isolated blood vessels from male and female Sham and TBI groups showing immunostaining for α 2A ‐AR (green), α 2B ‐AR (white), and the BMEC marker CD31 (red). Scale bar: 50 μm. (C) mRNA expression of Adra2a was significantly decreased in the isolated BMECs from the female TBI group ( n = 6 per group). (D) mRNA expression of Adra2b was significantly decreased in the isolated BMECs from the female TBI group compared with the female Sham and male TBI groups ( n = 6 per group). (E) α 2A ‐AR protein expression in BMECs was significantly reduced in blood vessels isolated from the female TBI group ( n = 4 per group). (F) α 2B ‐AR protein expression in BMECs was significantly reduced in blood vessels isolated from the female TBI group ( n = 4 per group). (G) Body weight gain was significantly reduced in the male and female TBI + saline groups compared with both male and female Sham and TBI + DEX groups ( n = 20 per group). (H) Grip strength was significantly decreased in both male and female TBI + saline groups compared with the male and female Sham and TBI + DEX groups ( n = 20 per group). (I) In the rotarod test, the latency to first fall was significantly decreased in both male and female TBI + saline groups, compared with the male and female Sham and TBI + DEX groups ( n = 20 per group). Statistical analysis was performed using two‐way ANOVA (sex [male, female], treatment [Sham, TBI + saline, TBI + DEX]) and Bonferroni's multiple‐comparison test. * p < 0.05; ** p < 0.01; **** p < 0.0001. Red annotations indicate sex‐related effects, whereas black annotations indicate treatment‐related effects.

Article Snippet: Magnetic cell separation was performed with mouse CD31 MicroBeads (catalog no. 130‐097‐418; Miltenyi Biotec, MD, USA) and a QuadroMACS Separator (Miltenyi Biotec, MD, USA) according to the manufacturer's instructions.

Techniques: Marker, Isolation, Staining, Immunostaining, Expressing, Saline, Analysis, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Sex‐Specific Effects of Dexmedetomidine on Brain Microvascular Endothelial Cell Function After Traumatic Brain Injury in Juvenile Mice

doi: 10.1002/cns.71204

Figure Lengend Snippet: Extravasation of Evans blue and tight junction protein evaluation. (A) Representative images of brains from Sham, TBI + saline, and TBI + DEX groups injected with Evans blue are shown for both males and females. (B) Evans blue concentrations were significantly elevated in tissue samples from injured brain regions in both male and female TBI + saline and TBI + DEX groups compared with Sham controls. Furthermore, Evans blue concentration was markedly higher in the TBI + saline group than in the TBI + DEX group ( n = 5 per group). (C) Representative images showing co‐localization of JAM‐A with the BMEC marker CD31 in isolated microvessels from Sham, TBI + saline and TBI + DEX groups in both males and females. CD31 (green; a BMEC marker); JAM‐A (red; a junctional adhesion protein); DAPI (blue; a nuclear marker). Scale bar: 10 μm. (D) Quantitative analysis of CD31 and JAM‐A co‐localization on isolated cerebral blood vessels ( n = 4 per group). Statistical analysis was performed using two‐way ANOVA (sex [male, female], treatment [Sham, TBI + saline, TBI + DEX]), and Bonferroni's multiple‐comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; n.s., not significant.

Article Snippet: Magnetic cell separation was performed with mouse CD31 MicroBeads (catalog no. 130‐097‐418; Miltenyi Biotec, MD, USA) and a QuadroMACS Separator (Miltenyi Biotec, MD, USA) according to the manufacturer's instructions.

Techniques: Saline, Injection, Tissue, Concentration Assay, Marker, Isolation, Analysis, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Sex‐Specific Effects of Dexmedetomidine on Brain Microvascular Endothelial Cell Function After Traumatic Brain Injury in Juvenile Mice

doi: 10.1002/cns.71204

Figure Lengend Snippet: Effect of DEX on BMEC function. The mRNA expression of Inos (A), Nos3 (B), Ocln (C), Cldn5 (D), Cfl1 (E), Pfn1 (F), Pecam1 (G), and Zic3 (H) in Sham, TBI + saline, and TBI + DEX groups in both male and female mice ( n = 6 per group). (I) Representative images of isolated blood vessels from male and female Sham, TBI + saline, and TBI + DEX groups showing immunostaining for occludin (green), VE‐cadherin (white), and the BMEC marker CD31 (red). Scale bar: 50 μm. (J) Co‐localization analysis of occludin protein expression with BMECs in isolated blood vessels ( n = 4 per group). (K) Co‐localization analysis of VE‐cadherin protein expression with BMECs in isolated blood vessels ( n = 4 per group). Statistical analysis was performed using two‐way ANOVA (sex [male, female], treatment [Sham, TBI + saline, TBI + DEX]) and Bonferroni's multiple‐comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Cfl1 , cofilin‐1; Cldn5 , claudin‐5; Inos , inducible nitric oxide synthase; Nos3 , nitric oxide synthase 3; Ocln , occludin; Pecam1 , platelet endothelial cell adhesion molecule 1; Pfn1 , profilin‐1; Zic3 , zic family member 3. Red annotations indicate sex‐related effects, whereas black annotations indicate treatment‐related effects.

Article Snippet: Magnetic cell separation was performed with mouse CD31 MicroBeads (catalog no. 130‐097‐418; Miltenyi Biotec, MD, USA) and a QuadroMACS Separator (Miltenyi Biotec, MD, USA) according to the manufacturer's instructions.

Techniques: Expressing, Saline, Isolation, Immunostaining, Marker, Analysis, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Sex‐Specific Effects of Dexmedetomidine on Brain Microvascular Endothelial Cell Function After Traumatic Brain Injury in Juvenile Mice

doi: 10.1002/cns.71204

Figure Lengend Snippet: Expression of proinflammatory markers. (A) Representative immunohistochemical images displaying co‐localization of CD31 (a BMEC marker; green) and TNF‐α (a proinflammatory marker; red) in isolated blood vessels obtained from male and female mice in the Sham, TBI + saline, and TBI + DEX groups. Scale bar: 50 μm. (B) Representative immunohistochemical images displaying co‐localization of ICAM‐1 (green) and CD31 (red) in isolated blood vessels obtained from male and female Sham, TBI + saline, and TBI + DEX treatment groups. Scale bar: 50 μm. (C–J) The mRNA expression of Tnf‐α (C), Il1b (D), Il6 (E), Tgfb1 (F), Tlr4 (G), Nlrp3 (H), Nfκb (I), and Icam1 (J) in the isolated BMECs from Sham, TBI + saline, and TBI + DEX groups in both male and female mice ( n = 6 mice per group). (K) Quantitative analysis of the co‐localization between CD31 and TNF‐α was performed on isolated blood vessels from male and female mice across Sham, TBI + saline, and TBI + DEX treatment groups ( n = 4 mice per group). (L) Quantitative analysis of the co‐localization between CD31 and ICAM‐1 was performed on isolated blood vessels from male and female Sham, TBI + saline, and TBI + DEX treatment groups ( n = 4 mice per group). Statistical analysis was performed using two‐way ANOVA (sex [male, female], treatment [Sham, TBI + saline, TBI + DEX]) and Bonferroni's multiple‐comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Icam‐1 , intercellular adhesion molecule 1; Il‐1β , interleukin‐1 beta; Il‐6 , interleukin‐6; Nfκb , nuclear factor kappa‐light‐chain‐enhancer of activated B cells; Nlrp3 , NOD‐, LRR‐, and pyrin domain‐containing protein 3; Tgf‐β1 , transforming growth factor beta 1; Tlr4 , toll‐like receptor 4; Tnf‐α , tumor necrosis factor‐alpha. Red annotations indicate sex‐related effects, whereas black annotations indicate treatment‐related effects.

Article Snippet: Magnetic cell separation was performed with mouse CD31 MicroBeads (catalog no. 130‐097‐418; Miltenyi Biotec, MD, USA) and a QuadroMACS Separator (Miltenyi Biotec, MD, USA) according to the manufacturer's instructions.

Techniques: Expressing, Immunohistochemical staining, Marker, Isolation, Saline, Analysis, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Sex‐Specific Effects of Dexmedetomidine on Brain Microvascular Endothelial Cell Function After Traumatic Brain Injury in Juvenile Mice

doi: 10.1002/cns.71204

Figure Lengend Snippet: Effects of DEX on ER stress responses. (A) Representative immunohistochemical images displaying co‐localization of phosphorylated IRE1α (green) and CD31 (red) in isolated blood vessels obtained from male and female Sham, TBI + saline, and TBI + DEX groups. Scale bar: 10 μm. (B) Representative images of CHOP expression. CHOP (green), CD31 (red; a BMEC marker), and DAPI (blue; a nuclear marker). Scale bar: 10 μm. (C) mRNA expression of Ire1α in the isolated BMECs ( n = 6 per experimental group). (D) mRNA expression of Perk in the isolated BMECs ( n = 6 per experimental group). (E) Quantitative analysis of the co‐localization between CD31 and phosphorylated IRE1α was performed on isolated blood vessels from male and female Sham, TBI + saline, and TBI + DEX treatment groups ( n = 4 mice per group). (F) CHOP protein expression was significantly higher in brain tissues from the TBI + saline groups ( n = 5 per experimental group). (G) Co‐localization analysis of CHOP and BMECs (CD31‐positive cells) in brain tissues across all experimental groups ( n = 5 per experimental group). Statistical analysis was performed using two‐way ANOVA (sex [male, female], treatment [Sham, TBI + saline, TBI + DEX]), and Bonferroni's multiple‐comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Magnetic cell separation was performed with mouse CD31 MicroBeads (catalog no. 130‐097‐418; Miltenyi Biotec, MD, USA) and a QuadroMACS Separator (Miltenyi Biotec, MD, USA) according to the manufacturer's instructions.

Techniques: Immunohistochemical staining, Isolation, Saline, Expressing, Marker, Analysis, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Sex‐Specific Effects of Dexmedetomidine on Brain Microvascular Endothelial Cell Function After Traumatic Brain Injury in Juvenile Mice

doi: 10.1002/cns.71204

Figure Lengend Snippet: Effect of DEX on ferroptosis. (A) Representative immunohistochemical images displaying co‐localization of GPX4 (green), 4‐HNE (white), and CD31 (red) in isolated blood vessels obtained from male and female Sham, TBI + saline, and TBI + DEX groups. Scale bar: 50 μm. (B) Representative immunohistochemical images displaying co‐localization of CD31 (green) and MDA (red) in isolated blood vessels obtained from male and female mice in the Sham, TBI + saline, and TBI + DEX groups. Scale bar: 50 μm. (C–F) The mRNA expression of Gpx4 (C), Ptgs2 (D), Hmox1 (E), and Tfrc (F) in isolated BMECs from Sham, TBI + saline, and TBI + DEX groups in both male and female mice ( n = 6 per experimental group). (G–I) Quantitative analyses of co‐localization between CD31 and GPX4 (G), 4‐HNE (H), or MDA (I) were performed on isolated blood vessels from male and female mice across Sham, TBI + saline, and TBI + DEX treatment groups ( n = 4 per experimental group). Statistical analysis was performed using two‐way ANOVA (sex [male, female], treatment [Sham, TBI + saline, TBI + DEX]), and Bonferroni's multiple‐comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Magnetic cell separation was performed with mouse CD31 MicroBeads (catalog no. 130‐097‐418; Miltenyi Biotec, MD, USA) and a QuadroMACS Separator (Miltenyi Biotec, MD, USA) according to the manufacturer's instructions.

Techniques: Immunohistochemical staining, Isolation, Saline, Expressing, Analysis, Comparison

Journal: Advanced Science

Article Title: Tumor‐Intrinsic HEXB Drives Endothelial‐To‐Mesenchymal Transition and Confers Resistance to Anti‐Angiogenic Therapy in Glioblastoma

doi: 10.1002/advs.78208

Figure Lengend Snippet: Glioma‐intrinsic HEXB promoted Endo‐MT changes of ECs. (A) Survival of GBM patients received bevacizumab treatment with different HEXB expression in GSE79671 (log‐rank test). (B) Survival of GBM patients received bevacizumab treatment with different HEXB expression in GSE72951 (log‐rank test). (C) The rate of response patients was lower in GBM patients with high HEXB expression in GSE72951 (Patients with a survival time superior to the median survival of the standard treatment were defined as the responder group, whereas those with a shorter survival time were defined as the non‐responder group). (D) The analysis of migration ability of HBMECs treated with different CM (sh#1: shHEXB#1; sh#2: shHEXB#2). (E) Permeability changes of HBMECs treated with different CM (sh#1: shHEXB#1; sh#2: shHEXB#2). (F) Immunofluorescence staining image of HBMECs treated with different CM (sh#1: shHEXB#1; sh#2: shHEXB#2). (G) Western blot results of HBMECs treated with different CM (sh#1: shHEXB#1; sh#2: shHEXB#2). (H) PCR results of HBMECs treated with different CM (sh#1: shHEXB#1; sh#2: shHEXB#2). (I) Flow cytometry of CD31 + cells in tumors from GSC1‐bearing mice. (J) Flow cytometry of CD31 + cells in tumors from GSC21‐bearing mice. (Date were presented as means ± standard deviations, * indicates p ≤ 0.05, ** indicates p < 0.01, **** indicates p < 0.0001.).

Article Snippet: The CD31 Microbeads (130‐097‐418, Miltenyi) were used for positive selection CD31 + endothelial cells from single‐cell suspensions of mouse tissues.

Techniques: Expressing, Analysis, Migration, Permeability, Immunofluorescence, Staining, Western Blot, Flow Cytometry

Journal: Bioactive Materials

Article Title: Injection site dictates the immune response to a biodegradable polymer and corresponding collagen regeneration

doi: 10.1016/j.bioactmat.2026.04.004

Figure Lengend Snippet: ID tissue exhibits higher densities of fibroblasts, macrophages, and microvessels compared to SC tissue. Immunofluorescence analysis comparing the expression of Vimentin (a fibroblast marker), CD68 (a macrophage marker), and CD31 (an endothelial cell marker) in ID and SC tissues. For each group, n = 3; data represent mean ± s.d.; ∗P < 0.05 and ∗∗P < 0.01.

Article Snippet: After antigen retrieval and blocking, sections were incubated overnight at 4 °C with primary antibodies targeting vimentin, CD68 (ABclonal, A20803), CD31 (R&D SYSTEMS, AF3628), HMGB1 (Cell Signaling Technology, 3935), HSP70 (ABclonal, A23457), NF-κB p65 (ABclonal, A19653), CD206 (Cell Signaling Technology, 24595), and FAPα (ABclonal, A23789 ).

Techniques: Immunofluorescence, Expressing, Marker