Journal: Molecular medicine reports

Article Title: Transdermal estrogen gel and oral aspirin combination therapy improves fertility prognosis via the promotion of endometrial receptivity in moderate to severe intrauterine adhesion.

doi: 10.3892/mmr.2018.8685

Figure Lengend Snippet: Figure 4. Fibrosis is significantly decreased in endometrium tissues within the combination therapy group following treatment. (A) Fibrosis was detected by Masson's Trichrome staining. Magnification, x200. Fibrotic areas were stained blue. (B) Expression of fibrosis markers (TGF‑β, CTGF, collagen I and collagen III) was detected by immunohistochemistry. Magnification, x200. The expression of fibrosis markers in endometrial tissues was determined at the mRNA level by (C) reverse transcription‑quantitative polymerase chain reactionand at the protein level by (D) western blotting. The relative quantity of proteins was calculated by comparison against the internal control (GAPDH) and noted below the bands. The postoperative tissues were obtained during the follow‑up at the 20‑24th day of the second menstrual cycle following transcervical resection of adhesion. Data are presented as the mean ± standard deviation. *P<0.05 vs. group A. CTGF, CCCTC‑binding factor; group A, transdermal estrogen gel therapy; group B, combination therapy of transdermal estrogen gel and oral aspirin therapy; TGF‑β, transforming growth factor‑β.

Article Snippet: The samples were blocked with normal goat serum (ZSGB‐Bio, Beijing, China) at 37 ̊C for 10 min. Then samples were incubated overnight at 4 ̊C with anti-rabbit transforming growth factor-β (TGF-β; 1:100; bs-0086R; BIOSS, Beijing, China), anti-rabbit CCCTC-binding factor (CTGF; 1:100; bs-0743R; BIOSS), anti-rabbit vascular endothelial growth factor (VEGF; 1:100; bs-1313R; BIOSS), anti-rabbit αvβ3 antibody (1:100; bs-0342R; BIOSS), anti-rabbit collagen I (COI; 1:100; bs-0578R; BIOSS), anti-rabbit collagen III (COIII; 1:100; bs‐0948R; BIOSS) or anti‐rabbit cluster of differentiation (CD31; 1:100; bs-20320R; BIOSS) primary antibodies.

Techniques: Staining, Expressing, Immunohistochemistry, Western Blot, Comparison, Control, Standard Deviation

Journal: Biomedicines

Article Title: AT2R Activation Improves Wound Healing in a Preclinical Mouse Model

doi: 10.3390/biomedicines12061238

Figure Lengend Snippet: AT2R activation during wound healing improves collagen I:III ratio by day 10. ( A ) Representative immunofluorescence images of wounded skin acquired at 20× stained for collagen 1 (green), collagen 3 (red), and DAPI (blue) and treated with saline (top left), C21 (top right), PD123319 (bottom left), and combination (bottom right). Scale = 100 µm. ( B ) Collagen I:III ratio (mean + SEM) at day 7 post-wounding. C21, PD123319, and combination groups have n = 8 replicates, while saline represents n = 24, all comparisons are ns, Kruskal–Wallis Test with multiple comparisons. ( C ) Collagen I:III ratio (mean + SEM) at day 10, * p = 0.0200 saline ( n = 23) vs. C21 ( n = 6), ** p = 0.0053 C21 vs. PD123319 ( n = 8), and ** p = 0.0041 C21 vs. combination ( n = 8), one-way ANOVA with multiple comparisons.

Article Snippet: For fluorescent visualization of collagen I (rabbit polyclonal anti-collagen I, Ab21286, 1:500, Abcam, Cambridge, UK), collagen 3 (rabbit polyclonal anti-collagen III, Ab7778, 1:100, Abcam, Cambridge, UK), CD31 (rabbit monoclonal anti-CD31/PECAM-1, clone D8V9E, 77699, 1:200, Cell Signaling, Danvers, MA, USA), CD45 (rabbit monoclonal anti-CD45, clone D3F8Q, 70257, 1:200, Cell Signaling, Danvers, MA, USA), Ly6G (rat anti-Ly6G, clone 1A8, 551459, 1:100, BD Pharmingen, San Diego, CA, USA), and F4/80 (rabbit monoclonal anti-F4-80, clone D2S9R, 70076, 1:100, Cell Signaling, Danvers, MA, USA), we employed multispectral multiplex immunofluorescence staining utilizing the Opal Manual Detection Kit (Akoya Biosciences, Marlborough, MA, USA), optimized for our tissue and targets of interest.

Techniques: Activation Assay, Immunofluorescence, Staining, Saline