Journal: BMC Medicine

Article Title: Coagulopathy triggered autoimmunity: experimental antiphospholipid syndrome in factor V Leiden mice

doi: 10.1186/1741-7015-11-92

Figure Lengend Snippet: Immunohistochemical staining for inflammatory and vascular markers in factor V Leiden (FVL) mice. Representative immunohistochemical staining images from the three groups: adjuvant-immunized FVL control (FVL-control), experimental antiphospholipid syndrome (eAPS), heterozygous FVL (FVL Q/+ -APS) and eAPS homozygous FVL (FVL Q/Q -APS) mice. Quantification data for each marker are also presented. (A–C,P) Glial fibrillary acidic protein (GFAP)-positive immunoreactions with similar expression in the area of the hippocampus (original magnification × 20). (D–F,Q) MAC3-positive cells (macrophages) in the meninges (black arrows) and in the parenchyma of the cortex (black arrowheads; original magnification × 20). (G–I,R) CD3-positive cells (T cells, black arrows; original magnification × 20). (J–L,S) Infiltrates with increased expression of B220-positive cells (B cells) in the control FVL group compared with the APS FVL Q/+ and APS FVL Q/Q groups (black arrows; original magnification × 40). (M–O,T) Representative images of vascular endothelial growth factor (VEGF) staining, with similar expression in the area of the cortex (original magnification × 20).

Article Snippet: After incubation of the sections in blocking buffer (Foetal bovine serum, FBS) they were treated with primary antibodies against glial acidic fibrillary protein (GFAP; Dako, Glostrup, Denmark), MAC3, B220 (both BD Biosciences, Inc., San Jose, CA, USA), CD3 (Neomarkers Inc., Fremont, CA, USA), vascular endothelial growth factor (VEGF; Spring Bioscience Corp., Pleasanton, CA, USA), for the detection of astrocytes, macrophage/microglia, B cells, T cells, and VEGF, respectively (dilutions: 1;500, 1:100, 1:100, 1:150, 1:100, respectively).

Techniques: Immunohistochemical staining, Staining, Adjuvant, Control, Marker, Expressing

Journal: International Journal of Nanomedicine

Article Title: The Dual Angiogenesis Effects via Nrf2/HO-1 Signaling Pathway of Melatonin Nanocomposite Scaffold on Promoting Diabetic Bone Defect Repair

doi: 10.2147/IJN.S449290

Figure Lengend Snippet: The angiogenesis properties of different scaffolds in vitro. ( a ) Scratch wound healing of b. End3 cultured with different scaffolds after 12 h, scale bar = 200 μm. ( b ) Statistical analysis of wound healing. ( c ) Tubule formation of b. End3, scale bar = 200 μm. ( d and e ) Quantitative analysis of nodes and total tube length. ( f ) Quantitative analysis of the angiogenesis-related gene expression. ( g – j ) The protein level of angiogenesis-related genes by Western blot analysis in b. End3 cells. ( k ) ELISA quantitative results of cytokine PDGFA secreted from VECs. ( l and m ) Immunofluorescence images of Ang2, CD31, and VEGF staining (scale bar: 50 μm) and relative quantitative analysis of fluorescence intensity. ( n – q ) Activity of lipid oxidation metabolites MAD, antioxidant enzymes SOD, GSH, and hydroxyl radical scavenging rate of b. End3. n=3, *p < 0.05, **p < 0.01.

Article Snippet: MC3T3-E1 and bEnd.3 cells and different groups of scaffolds were cultured for 48 h. The cells were washed with PBS and subsequently fixed with paraformaldehyde (4%) and incubated with primary antibodies vinculin (Sigma, Cat#V9131), NF-E2–related factor 2 (Nrf2, Cell Signaling Technology, Cat#12,721), vascular endothelial growth factor (VEGF, HUABIO, Cat#HN0727), cluster of differentiation 31 (CD31, Abcam, Cat # ab28364), angiopoietin 2 (Ang2, HUABIO, Cat#RT1046) overnight at 4°C.

Techniques: In Vitro, Cell Culture, Gene Expression, Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Fluorescence, Activity Assay

Journal: International Journal of Nanomedicine

Article Title: The Dual Angiogenesis Effects via Nrf2/HO-1 Signaling Pathway of Melatonin Nanocomposite Scaffold on Promoting Diabetic Bone Defect Repair

doi: 10.2147/IJN.S449290

Figure Lengend Snippet: The mechanism of MT nanocomposite scaffold to promote angiogenesis. ( a ) The DCF fluorescence images represented the ROS level, scale bar: 50 μm. ( b ) Quantitative analysis of the relative DCF fluorescence intensity. ( c ) qPCR analysis of the mRNA expression levels of intracellular antioxidant and angiogenesis-related genes. ( d – g ) Immunofluorescence images of the Nrf2 and VEGF staining (scale bar: 50 μm) and relative quantitative analysis of fluorescence intensity. ( h – j ) The protein level of Nrf2 and VEGF by Western blot analysis in bEend.3 cell. ( k ) ELISA quantitative results of cytokine VEGF secreted from VECs. ( l ) Schematic diagram showing the mechanism of Nrf2-mediated angiogenesis activated by MT nanocomposite scaffold. n=3; *p < 0.05, **p < 0.01.

Article Snippet: MC3T3-E1 and bEnd.3 cells and different groups of scaffolds were cultured for 48 h. The cells were washed with PBS and subsequently fixed with paraformaldehyde (4%) and incubated with primary antibodies vinculin (Sigma, Cat#V9131), NF-E2–related factor 2 (Nrf2, Cell Signaling Technology, Cat#12,721), vascular endothelial growth factor (VEGF, HUABIO, Cat#HN0727), cluster of differentiation 31 (CD31, Abcam, Cat # ab28364), angiopoietin 2 (Ang2, HUABIO, Cat#RT1046) overnight at 4°C.

Techniques: Fluorescence, Expressing, Immunofluorescence, Staining, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Nanomedicine

Article Title: The Dual Angiogenesis Effects via Nrf2/HO-1 Signaling Pathway of Melatonin Nanocomposite Scaffold on Promoting Diabetic Bone Defect Repair

doi: 10.2147/IJN.S449290

Figure Lengend Snippet: Evaluation of in vivo bone defect repair following scaffold implantation through histological and immunofluorescence staining. ( a ) Images of H&E and Masson trichrome staining in the skull defect area at 4 and 8 weeks postoperatively. ( b – d ) Immunofluorescence staining of Nrf2 and VEGF in the skull defect area at 8 weeks after implantation. n=3; *p < 0.05, **p < 0.01.

Article Snippet: MC3T3-E1 and bEnd.3 cells and different groups of scaffolds were cultured for 48 h. The cells were washed with PBS and subsequently fixed with paraformaldehyde (4%) and incubated with primary antibodies vinculin (Sigma, Cat#V9131), NF-E2–related factor 2 (Nrf2, Cell Signaling Technology, Cat#12,721), vascular endothelial growth factor (VEGF, HUABIO, Cat#HN0727), cluster of differentiation 31 (CD31, Abcam, Cat # ab28364), angiopoietin 2 (Ang2, HUABIO, Cat#RT1046) overnight at 4°C.

Techniques: In Vivo, Immunofluorescence, Staining