Journal: Reproduction (Cambridge, England)

Article Title: Estradiol promotes cells invasion by activating β-catenin signaling pathway in endometriosis

doi: 10.1530/REP-15-0371

Figure Lengend Snippet: (A, B, C and D) Effects of E 2 and E 2 +ICI on the β-catenin expression. (A and C) Dose-dependence by the stimulation with E 2 . HESCs were cultured in a phenol red-free DMEM/F12 for 24 h and incubated with various concentrations of E 2 and E 2 +ICI for 24 h. (B and D) Time course by the stimulation with E 2 . Cells were cultured in a phenol red-free DMEM/F12 for 24 h and treated with E 2 (10 −8 mol/l), E 2 +ICI for the indicated times (0, 12, 24, and 48 h). β-catenin mRNA was extracted by TRIzol and examined by RT-PCR (A and B). Protein samples were separated by 10% SDS–PAGE and subjected to western blot analysis for total β-catenin (C and D). (E) Effects of E 2 on the nuclear localization of β-catenin in HESCs. Cells were stimulated with DMSO or E 2 (10 −8 mol/l) for 48 h, followed by western blot, and the cytoplasmic and nucleus of β-catenin was observed. (F) Effects of E 2 on the non-phosphorylated β-catenin (active β-catenin) in HESCs. Cells were stimulated with DMSO or E 2 (10 −8 mol/l) for 48 h, followed by western blot, and non-phosphorylated β-catenin was observed. (G) E 2 increases TCF transcriptional activity. TOPflash or FOPflash was co-transfected with pRL-SV40 into HESCs. After transfection for 24 h, the cells were stimulated with E 2 (10 −8 mol/l) and E 2 plus ICI for 48 h. The luciferase activities are shown as percentages of the control level. Data are expressed as mean± s.e.m . * P <0.05 and ** P <0.01 vs controls. Data presented are from three independent experiments.

Article Snippet: Appropriate amounts of mouse-directed antibody against β-catenin (sc-133239, Santa Cruz Biotechnology), mouse antibody directed against estrogen receptor alpha (ESR1; sc-73479, Santa Cruz Biotechnology), and mouse nonspecific IgG (Biosense, Bergen, Norway) were then added and incubated for 4 h at 4 °C.

Techniques: Expressing, Cell Culture, Incubation, Reverse Transcription Polymerase Chain Reaction, SDS Page, Western Blot, Activity Assay, Transfection, Luciferase

Journal: Reproduction (Cambridge, England)

Article Title: Estradiol promotes cells invasion by activating β-catenin signaling pathway in endometriosis

doi: 10.1530/REP-15-0371

Figure Lengend Snippet: (A) The interaction between ESR1 and β-catenin in HESCs was examined. Cells were incubated with DMSO and E 2 (10 −8 mol/l) for 24 h. Then cell lysates were prepared and immunoprecipitated with IgG and ESR1 antibody. The second panel is ESR1 shown as an internal loading control. Immunoblots of input lysate controls (5% of input) are also shown. (B and C) HESCs were incubated with E 2 (10 −8 mol/l) under different time scales (0, 12, 24, and 48 h). (B) Cell lysates were prepared and immunoprecipitated with ESR1 antibody or IgG. The interaction between ESR1 and β-catenin/TCF3/LEF1 was examined. (C) Cell lysates were prepared and immunoprecipitated with β-catenin antibody or IgG. The interaction between β-catenin and ESR1/TCF3/LEF1 was examined. (D and E) Co-localization of ESR1, β-catenin, and LEF1/TCF3 in nucleus were stimulated with E 2 in HESCs. Cells were cultured in phenol red-free DMEM/F12 for 24 h and incubated with DMSO and E 2 (10 −8 mol/l) for 48 h. (D) Representative confocal microscopy images of HESCs immunostained for ERS1 (green, left panels) and its co-localization with the LEF1 and TCF3 (red, middle panels), and the merged images (right panels). (E) Representative confocal microscopy images of HESCs immunostained for β-catenin (green, left panels) and its co-localization with the LEF1 and TCF3 (red, middle panels), and the merged images (right panels). The bars indicate 20 μm. Data presented are from three independent experiments.

Article Snippet: Appropriate amounts of mouse-directed antibody against β-catenin (sc-133239, Santa Cruz Biotechnology), mouse antibody directed against estrogen receptor alpha (ESR1; sc-73479, Santa Cruz Biotechnology), and mouse nonspecific IgG (Biosense, Bergen, Norway) were then added and incubated for 4 h at 4 °C.

Techniques: Incubation, Immunoprecipitation, Western Blot, Cell Culture, Confocal Microscopy

Journal: Reproduction (Cambridge, England)

Article Title: Estradiol promotes cells invasion by activating β-catenin signaling pathway in endometriosis

doi: 10.1530/REP-15-0371

Figure Lengend Snippet: (A) Effect of E 2 on VEGF and MMP9 expression. Cells were stimulated with vehicle, E 2 (10 −8 mol/l), and E 2 (10 −8 mol/l)+ICI for 48 h. β-catenin, MMP9, and VEGF mRNA were detected by qRT-PCR. (B) Effect of β-catenin siRNA on VEGF and MMP9 expression. After β-catenin, siRNA was transfected for 24 h, the cells were stimulated with vehicle and E 2 (10 −8 mol/l) for 48 h. β-catenin, MMP9, and VEGF were detected by qRT-PCR. (C) E 2 enhances the invasive ability of HESCs. Left: representative photomicrographs of invasion of vehicle, E 2 , and E 2 +ICI-treated HESCs. Right: number of invasive cells/mm 2 in DMSO, E 2 , and E 2 +ICI-treated HESCs. (D) Effect of β-catenin deletion on cells invasiveness ability stimulation with E 2 . Left: representative photomicrographs of invasion of control, β-catenin siRNA, or scrambled siRNA transfected HESCs. Right: number of invasive cells/mm 2 in control, β-catenin siRNA, or scrambled siRNA transfected HESCs. Data are expressed as mean± s.e.m . * P <0.05 vs controls. Data presented are from three independent experiments.

Article Snippet: Appropriate amounts of mouse-directed antibody against β-catenin (sc-133239, Santa Cruz Biotechnology), mouse antibody directed against estrogen receptor alpha (ESR1; sc-73479, Santa Cruz Biotechnology), and mouse nonspecific IgG (Biosense, Bergen, Norway) were then added and incubated for 4 h at 4 °C.

Techniques: Expressing, Quantitative RT-PCR, Transfection

Journal: Reproduction (Cambridge, England)

Article Title: Estradiol promotes cells invasion by activating β-catenin signaling pathway in endometriosis

doi: 10.1530/REP-15-0371

Figure Lengend Snippet: The flow chart and representative pictures of the endometriotic animal model. (A) Fifty NOD–SCID female mice transplanted with human normal endometrium were separated into six groups: mice treated with E 2 plus positive β-catenin siRNA or scrambled siRNA and saline solution control after a 10-day incubation; mice treated with E 2 plus positive β-catenin siRNA or scrambled siRNA and saline solution control after a 21-day incubation. (B and C) Representative pictures of the implanted fragments of endometrial tissues into the peritoneal cavity of the NOD–SCID mice after transplantation of normal endometrium. The arrows indicate the implanted endometrial tissues. (D) The murine lesions were examined by histopathology. (E) Western blotting analysis of non-phosphorylated β-catenin and ESR1 expression of pre-transplanted tissues and endometriotic lesions obtained from mice model (a 10-day incubation). (F) Western blotting analysis of non-phosphorylated β-catenin and ESR1 expression of pre-transplanted tissues and endometriotic lesions obtained from mice model (a 21-day incubation). H&E, hematoxylin and eosin stain.

Article Snippet: Appropriate amounts of mouse-directed antibody against β-catenin (sc-133239, Santa Cruz Biotechnology), mouse antibody directed against estrogen receptor alpha (ESR1; sc-73479, Santa Cruz Biotechnology), and mouse nonspecific IgG (Biosense, Bergen, Norway) were then added and incubated for 4 h at 4 °C.

Techniques: Animal Model, Incubation, Transplantation Assay, Histopathology, Western Blot, Expressing, H&E Stain

Journal: Journal of Food and Drug Analysis

Article Title: Cannabis sativa L .: A comprehensive review on legislation, decriminalization, phytochemistry, antimicrobial activity, and safety

doi: 10.38212/2224-6614.3471

Figure Lengend Snippet: Antibacterial activity of Cannabis sativa volatile and organic extracts.

Article Snippet: Last but not least, the “Futura” cultivars showed high to moderate activity against the various tested bacteria, with MIC values ranging from 0.625 μL/mL to >20 μL/mL against Enterococcus faecium ATCC 19434, which was consistent with earlier investigations [ ].

Techniques: Activity Assay, Cannabis, Diffusion-based Assay, Inhibition, Bacteria, Antibiofilm Assay, Serial Time-encoded Amplified Microscopy, Distillation, Drug Susceptibility Assay

Journal: Journal of Food and Drug Analysis

Article Title: Cannabis sativa L .: A comprehensive review on legislation, decriminalization, phytochemistry, antimicrobial activity, and safety

doi: 10.38212/2224-6614.3471

Figure Lengend Snippet: Antibacterial activity of Cannabis sativa bioactive compounds.

Article Snippet: Last but not least, the “Futura” cultivars showed high to moderate activity against the various tested bacteria, with MIC values ranging from 0.625 μL/mL to >20 μL/mL against Enterococcus faecium ATCC 19434, which was consistent with earlier investigations [ ].

Techniques: Activity Assay, Cannabis, Staining, Membrane, Diffusion-based Assay, Drug Susceptibility Assay, Motility Assay, Tube Formation Assay, Infection, Inhibition, Bacteria

Journal: Gastrohep

Article Title: SARS‐CoV‐2 vaccination in patients with inflammatory bowel disease

doi: 10.1002/ygh2.473

Figure Lengend Snippet: Vaccines with regulatory approval or undergoing Phase 3 trials

Article Snippet: A pre‐print article reported that the AstraZeneca vaccine (ChAdOx1‐nCoV19) efficacy against mild‐moderate COVID‐19 from the B.1.351 South African escape variant was only 10.4% compared to an efficacy of 75% for non‐B.1.351 variants.

Techniques: Vaccines, Formulation, Plasmid Preparation, Expressing, Infection, Biomarker Discovery, Recombinant, Adjuvant