Journal: International Journal of Neuropsychopharmacology

Article Title: Mesenchymal Stem Cell–Derived Extracellular Vesicles Alleviate M1 Microglial Activation in Brain Injury of Mice With Subarachnoid Hemorrhage via microRNA-140-5p Delivery

doi: 10.1093/ijnp/pyab096

Figure Lengend Snippet: Activin-like kinase 5 (ALK5) silencing reduces NADPH oxidase 2 (NOX2) expression to contribute to suppression of M1 microglia activation in subarachnoid hemorrhage (SAH) mice. A, Quantification of CD16-positive M1 microglia and CD206-positive M2 microglia. B, Western blot analysis of NOX2 protein expression in SAH mice after silencing ALK5. * P <.05 vs sham-operated mice; # P <.05 vs SAH mice treated with shRNA negative control (sh-NC). C, NOX2 knockdown efficiency confirmed by western blot analysis along with quantification of CD16-positive M1 microglia and CD206-positive M2 microglia. * P <.05 vs wild type (WT) mice. D, Western blot analysis of ALK5 protein expression in NOX2−/− mice pretreated with oe-ALK5 lentiviral vector. E, Quantification of CD16-positive M1 microglia and CD206-positive M2 microglia. n = 6. * P <.05 vs NOX2−/− mice pretreated with vector. The data were measurement data, which were expressed by mean ± standard deviation. The comparison between the two groups was analyzed by unpaired t-test. Comparison among multiple groups was analyzed by one-way variance of analysis.

Article Snippet: Then, sections/cells were incubated with primary antibodies to CD16 (1:200, A13980, ABclonal), CD206 (1:100, ab195191, Abcam), and Iba 1 (1:100, A12391, ABclonal) overnight at 4°C, and incubated with corresponding fluorescent secondary antibodies (AS011 and AS007, ABclonal).

Techniques: Expressing, Activation Assay, Western Blot, shRNA, Negative Control, Knockdown, Plasmid Preparation, Standard Deviation, Comparison

Journal: International Journal of Neuropsychopharmacology

Article Title: Mesenchymal Stem Cell–Derived Extracellular Vesicles Alleviate M1 Microglial Activation in Brain Injury of Mice With Subarachnoid Hemorrhage via microRNA-140-5p Delivery

doi: 10.1093/ijnp/pyab096

Figure Lengend Snippet: miR-140-5p shuttled by MSC-derived EVs represses brain injury and microglial M1 activation in mice after subarachnoid hemorrhage (SAH). A, The expression of miR-140-5p in EVs from MSCs transfected with miR-140-5p mimic or miR-140-5p inhibitor detected by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). * P <.05 vs EVs from mimic-NC-transfected MSCs. # P <.05 vs EVs from inhibitor-NC-transfected MSCs. Sham-operated mice were used as control, and SAH mice were treated or not treated with EVs or EVs-miR-140-5p. B, Quantification of neurological function score (n = 6). C, TdT-mediated dUTP-biotin nick end labeling with NeuN (TUNEL/NeuN) immunofluorescence staining of the cerebral cortex (scale bar = 100 μm; DAPI: blue; TUNEL: green; NeuN: red) and the number of the TUNEL/NeuN positive cells (n = 6). D, The expression of IL-6, IL-1β, and TNF-α in serum detected by enzyme-linked immunosorbent assay (ELISA) (n = 6). E, Representative immunofluorescence micrographs of microglia polarization the cerebral cortex and the number of CD16-positive M1 microglia and CD206-positive M2 microglia (scale bar = 25 μm; CD16/CD206: red, Iba 1: green, DAPI: blue). F, The expression of miR-140-5p in SAH mice detected by RT-qPCR. G, Western blot analysis of the expression of ALK5 and NADPH oxidase 2 (NOX2) in SAH mice. * P <.05 vs SAH mice; # P <.05 vs SAH mice treated with EVs. The data were measurement data, which were expressed by mean ± standard deviation. Comparison among multiple groups was analyzed by one-way variance of analysis. n = 6..

Article Snippet: Then, sections/cells were incubated with primary antibodies to CD16 (1:200, A13980, ABclonal), CD206 (1:100, ab195191, Abcam), and Iba 1 (1:100, A12391, ABclonal) overnight at 4°C, and incubated with corresponding fluorescent secondary antibodies (AS011 and AS007, ABclonal).

Techniques: Derivative Assay, Activation Assay, Expressing, Transfection, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Control, End Labeling, TUNEL Assay, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation, Comparison

Journal: International Journal of Neuropsychopharmacology

Article Title: Mesenchymal Stem Cell–Derived Extracellular Vesicles Alleviate M1 Microglial Activation in Brain Injury of Mice With Subarachnoid Hemorrhage via microRNA-140-5p Delivery

doi: 10.1093/ijnp/pyab096

Figure Lengend Snippet: miR-140-5p shuttled by MSC-EVs diminishes Activin-like kinase 5 (ALK5) expression to suppress brain injury and microglial M1 activation in subarachnoid hemorrhage (SAH) mice. SAH mice were treated with EVs, EVs-miR-140-5p, or EVs-miR-140-5p + ALK5. A, The expression of IL-6, IL-1β, and TNF-α in serum detected by enzyme-linked immunosorbent assay (ELISA) (n = 6). B, The number of CD16-positive M1 microglia and CD206-positive M2 microglia. C, The expression of miR-140-5p in SAH mice detected by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). D, Western blot analysis of the expression of ALK5 and NADPH oxidase 2 (NOX2) in SAH mice. * P <.05 vs SAH mice treated with EVs; # P <.05 vs SAH mice treated with EVs-miR-140-5p. The data were measurement data, which were expressed by mean ± standard deviation. Comparison among multiple groups was analyzed by one-way variance of analysis. n = 6..

Article Snippet: Then, sections/cells were incubated with primary antibodies to CD16 (1:200, A13980, ABclonal), CD206 (1:100, ab195191, Abcam), and Iba 1 (1:100, A12391, ABclonal) overnight at 4°C, and incubated with corresponding fluorescent secondary antibodies (AS011 and AS007, ABclonal).

Techniques: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Standard Deviation, Comparison

Journal: Antioxidants

Article Title: Superoxide-Mediated Upregulation of MMP9 Participates in BMPR2 Destabilization and Pulmonary Hypertension Development

doi: 10.3390/antiox12111961

Figure Lengend Snippet: ( a ) Western blot analysis for BMPR2 expression in lung tissues from wild-type and MMP9 KO mice exposed to normoxia or Sugen plus hypoxia. ( b ) Western blot analysis for COMP expression in lung tissues from wild-type and MMP9 KO mice exposed to normoxia or Sugen plus hypoxia. ( n = 4–7) * p < 0.05 vs. Nor (WT) using one-way ANOVA with Tukey’s multiple comparison test.

Article Snippet: Specific antibodies were purchased from the companies indicated: anti-MMP9 (Abcam, Cambridge, MA, USA: ab38898), anti-BMPR2 (Abclonal, Woburn, MA, USA: A16778), anti-COMP (Abclonal, Woburn, MA, USA: A13963) anti-GC (Sigma, Burlington, MA, USA: G4405), anti-SIRT3 (Abcam, Cambridge, MA: ab86671), anti-SOD2 (Abclonal, Woburn, MA, USA: A1340) and anti-β-actin (Sigma, Burlington, MA, USA: A5441).

Techniques: Western Blot, Expressing

Journal: World Journal of Gastroenterology

Article Title: Nimbolide inhibits tumor growth by restoring hepatic tight junction protein expression and reduced inflammation in an experimental hepatocarcinogenesis

doi: 10.3748/wjg.v26.i45.7131

Figure Lengend Snippet: Effect of nimbolide on liver function parameters and tumor and cell proliferation markers in diethylnitrosamine and N-nitrosomorpholine induced hepatocellular carcinoma mice. A: Plasma aspartate aminotransferase level in naïve and experimental groups; B: Plasma alanine aminotransferase level in naïve and experimental groups; C: Plasma alkaline phosphatase level in naïve and experimental groups; D: Plasma alpha-fetoprotein level in naïve and experimental groups; E: Representative images of glypican-3 immunostaining of mice liver from naive and experimental groups (200 × magnification); F: Quantification of glypican-3 positive cells in experimental groups by counting five 400 × fields of each liver section; G: Representative images of proliferating hepatocytes by proliferating cell nuclear antigen (PCNA) immunostaining of mice liver from naive and experimental groups (200 × magnification); H: Quantification of PCNA positive nuclei in experimental groups by counting five 400 × fields of each liver section. Scale bar: 200 μm; all the data are expressed as mean ± SEM ( n = 3-6). The comparison between the groups was analyzed by one-way ANOVA followed by Tukey’s multiple comparison post-hoc test or Kruskal-Wallis followed by Dunn’s multiple comparison post-hoc test. c P < 0.0001 compared to naïve group; d P < 0.05, e P < 0.01 compared to hepatocellular carcinoma group. AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; ALP: Alkaline phosphatase; AFP: Alpha-fetoprotein; PCNA: Proliferating cell nuclear antigen; HPF: High-power fields; HCC: Hepatocellular carcinoma.

Article Snippet: Primary antibodies used were rabbit polyclonal anti-glypican-3 (1:100, A13988; Abclonal, Woburn, MA, United States), rabbit polyclonal anti-PCNA (1:100, A0264; Abclonal, Woburn, MA, United States), and rabbit polyclonal anti-4-Hydroxynonenal (4HNE) (1:100, bs-6313R; Bioss, Woburn, Massachusetts, United States).

Techniques: Immunostaining