Journal: Translational Pediatrics

Article Title: Identification and evaluation of fructose-bisphosphate aldolase B as a potential diagnostic biomarker in choledochal cysts patients: a quantitative proteomic analysis

doi: 10.21037/tp-21-336

Figure Lengend Snippet: Histopathologic and expression profile changes in the CC tissues. (A) Real-time polymerase chain reaction (PCR) analysis of biomarker candidates in human tissues. *P<0.05 vs. control group, **P<0.01 vs. control group, and ***P<0.001 vs. control group. (B) Representative Western blot analysis of ALDOB protein expression in human tissues. (C,D) Representative image of Hematoxylin and Eosin and immunohistochemical staining of ALDOB in human tissues. Bar =100 µm. CC, choledochal cyst; IGFBP2, insulin-like growth factor binding protein 2; TPM3, tropomyosin; ALDOB, fructose-bisphosphate aldolase B; FAH, fumarylacetoacetate hydrolase; SOD3, superoxide dismutase 3; SPARC, secreted protein acidic and cysteine rich; APOE, apolipoprotein E; RBP4, retinol binding protein 4.

Article Snippet: The primary antibodies were anti-IGFBP2 (ab227881, 1:1,000, Abcam, USA), anti-TPM3 (ab190993, 1:1,000, Abcam, USA), anti-ALDOB (ab129728, 1:1,000, Abcam, USA), anti-FAH (PA5-42049, 1:1,000, Invitrogen, USA), anti-SOD3 (14316-1-AP, 1:1,000, Proteintech, China), anti-SPARC (ab55847, 1:1,000, Abcam, USA), anti-APOE (ab52607, 1:1,000, Abcam, USA), anti-RBP4 (ab154914, 1:1,000, Abcam, USA), and anti-GAPDH (ab8245, 1:5,000, Abcam, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Biomarker Assay, Western Blot, Immunohistochemical staining, Staining, Binding Assay

Journal: Pulmonary Circulation

Article Title: Hydrocortisone normalizes phosphodiesterase-5 activity in pulmonary artery smooth muscle cells from lambs with persistent pulmonary hypertension of the newborn

doi: 10.1086/674903

Figure Lengend Snippet: Hydrocortisone (HC) increases expression of extracellular superoxide dismutase (ecSOD) in fetal pulmonary artery smooth muscle cells (FPASMCs) from lambs with persistent pulmonary hypertension of the newborn (PPHN). A, PPHN FPASMC were exposed to 21% O2–5% CO2 or 95% O2–5% CO2 ± 100 nM HC for 24 hours. Cells were harvested for total protein and subjected to ecSOD Western blot analysis, with β-actin normalization. Data are shown as means ± SEM (n = 7 for 21% untreated, 21% plus 100 nM HC, 95% untreated, n = 8 for 95% plus 100 nM HC). Number sign indicates P < 0.05 versus 95% untreated. B, Representative Western blots are shown for ecSOD and β-actin in PPHN FPASMC.

Article Snippet: 13 Membranes were blocked for 1 hour at room temperature with 5% nonfat dry milk in Tris-buffered saline containing 0.1% Tween 20 (1X TBST) and incubated overnight at 4°C with primary antibody in 5% milk plus 1X TBST at an appropriate dilution (1:500 for mouse anti-PDE5 [BD Transduction], 1∶500 for goat anti-IκB [Santa Cruz Biotechnology], 1∶1,000 for rabbit anti-Nox1 [Santa Cruz Biotechnology], 1∶1000 for rabbit anti-Nox4 [Santa Cruz Biotechnology], 1∶1,000 for rabbit anti-extracellular superoxide dismutase [ecSOD; Enzo Life Sciences], and 1∶2,000 for mouse β-actin [Sigma]).

Techniques: Expressing, Western Blot

Journal: Pulmonary Circulation

Article Title: Hydrocortisone normalizes phosphodiesterase-5 activity in pulmonary artery smooth muscle cells from lambs with persistent pulmonary hypertension of the newborn

doi: 10.1086/674903

Figure Lengend Snippet: Model for the effects of hydrocortisone on phosphodiesterase-5 (PDE5) in fetal pulmonary artery smooth muscle cells (FPASMCs) from lambs with persistent pulmonary hypertension of the newborn (PPHN). Exposure to hyperoxia leads to increased reactive oxygen species (ROS) production, resulting in increased nuclear factor κ B (NFκB) activity with subsequent induction of PDE5 activity. Hydrocortisone disrupts this pathway by increasing extracellular superoxide dismutase (ecSOD) expression, leading to attenuation of ROS signaling and normalization of PDE5 activity. IκB: NFκB inhibitory protein.

Article Snippet: 13 Membranes were blocked for 1 hour at room temperature with 5% nonfat dry milk in Tris-buffered saline containing 0.1% Tween 20 (1X TBST) and incubated overnight at 4°C with primary antibody in 5% milk plus 1X TBST at an appropriate dilution (1:500 for mouse anti-PDE5 [BD Transduction], 1∶500 for goat anti-IκB [Santa Cruz Biotechnology], 1∶1,000 for rabbit anti-Nox1 [Santa Cruz Biotechnology], 1∶1000 for rabbit anti-Nox4 [Santa Cruz Biotechnology], 1∶1,000 for rabbit anti-extracellular superoxide dismutase [ecSOD; Enzo Life Sciences], and 1∶2,000 for mouse β-actin [Sigma]).

Techniques: Activity Assay, Expressing

Journal: Veterinary Medicine and Science

Article Title: Expression of xenobiotic metabolising enzymes in lungs of horses with or without histological evidence of lower airway inflammation

doi: 10.1002/vms3.331

Figure Lengend Snippet: Accession numbers and primer sequences for CYP1A1, CYP2A13, GSTM1, GSTP1, SOD3 and GAPDH respectively

Article Snippet: Primary SOD3 rabbit anti‐human/mouse/rat , Biosite, USA , 1:3,000.

Techniques:

Journal: Veterinary Medicine and Science

Article Title: Expression of xenobiotic metabolising enzymes in lungs of horses with or without histological evidence of lower airway inflammation

doi: 10.1002/vms3.331

Figure Lengend Snippet: Polyclonal antibodies used in the immunohistochemistry

Article Snippet: Primary SOD3 rabbit anti‐human/mouse/rat , Biosite, USA , 1:3,000.

Techniques: Plasmid Preparation

Journal: Veterinary Medicine and Science

Article Title: Expression of xenobiotic metabolising enzymes in lungs of horses with or without histological evidence of lower airway inflammation

doi: 10.1002/vms3.331

Figure Lengend Snippet: Box plots showing the level of gene expression of cytochrome P450 (CYP) enzymes (a) CYP1A1 and (b) CYP2A13, and enzymes involved in defence against toxic metabolites glutathione‐S‐transferases, (c) GSTM1, (d) GSTP1 and superoxide dismutase, (e) SOD3 in the lungs of horses without (Histo−) or with (Histo+) histological evidence of lower airway inflammation respectively. Gene expression is related to expression of the reference gene GAPDH (according to Schmittgen & Livak, ). The central horizontal line in each box represents the median, whereas the top and bottom lines represent the 25th and 75th percentiles. Whiskers indicate the min and max. p ‐values are shown above each enzyme, a p < .05 indicate a significant difference between horses without or with histological evidence of lower airway inflammation

Article Snippet: Primary SOD3 rabbit anti‐human/mouse/rat , Biosite, USA , 1:3,000.

Techniques: Gene Expression, Expressing

Journal: Veterinary Medicine and Science

Article Title: Expression of xenobiotic metabolising enzymes in lungs of horses with or without histological evidence of lower airway inflammation

doi: 10.1002/vms3.331

Figure Lengend Snippet: Immunohistochemical staining (brown) of (a) CYP1A, (b) SOD3, (c) GSTM1 and (d) GSTP1 in paraffin sections of horse lung tissue. ec, epithelial cells; lbs, lumen of bronchus; lb, lumen of bronchioles; lp, lamina propria; lrb, lumen of respiratory bronchioles; ltb, lumen of terminal bronchioles. Bars = 50 µm

Article Snippet: Primary SOD3 rabbit anti‐human/mouse/rat , Biosite, USA , 1:3,000.

Techniques: Immunohistochemical staining, Staining