Journal: Clinical and Translational Science

Article Title: Safety, Tolerability, and Pharmacokinetics of Single and Multiple Topical Applications of Sodium Taurodeoxycholate, a Treatment for Atopic Dermatitis

doi: 10.1111/cts.70242

Figure Lengend Snippet: Mean baseline plasma concentration‐time profiles of sodium taurodeoxycholate (TDCA) by treatment groups before the sodium TDCA administration at (a) single‐ascending dose and (b) multiple‐ascending dose study.

Article Snippet: Sodium taurodeoxycholate (TDCA) gel (NuGel) is a topical formulation of TDCA, a taurine‐conjugated bile acid derivative developed by Shaperon Inc. (Seoul, Republic of Korea) as a novel therapeutic candidate for AD.

Techniques: Clinical Proteomics, Concentration Assay

Journal: Clinical and Translational Science

Article Title: Safety, Tolerability, and Pharmacokinetics of Single and Multiple Topical Applications of Sodium Taurodeoxycholate, a Treatment for Atopic Dermatitis

doi: 10.1111/cts.70242

Figure Lengend Snippet: Mean plasma concentration‐time profiles of sodium taurodeoxycholate (TDCA) over 24 h by treatment groups at (a) single‐ascending dose and (b) multiple‐ascending dose study.

Article Snippet: Sodium taurodeoxycholate (TDCA) gel (NuGel) is a topical formulation of TDCA, a taurine‐conjugated bile acid derivative developed by Shaperon Inc. (Seoul, Republic of Korea) as a novel therapeutic candidate for AD.

Techniques: Clinical Proteomics, Concentration Assay

Journal: iScience

Article Title: A physiologically based model of bile acid metabolism in mice

doi: 10.1016/j.isci.2023.107922

Figure Lengend Snippet: Physicochemical properties of bile acids used

Article Snippet: tDCA , pKa (basic) , −0.2 , ChemAxon (HMDB, ) .

Techniques: Solubility

Journal: Frontiers in Immunology

Article Title: USP50 regulates NLRP3 inflammasome activation in duodenogastric reflux-induced gastric tumorigenesis

doi: 10.3389/fimmu.2024.1326137

Figure Lengend Snippet: Macrophage pyroptosis occurred in human bile reflux gastritis and experimental models. (A-D) Caspase-1 (A) , NLRP3 (B) , IL-1β (C) , and IL-18 (D) were analyzed via Immunofluorescence staining (red) which was performed on paraffin sections of non-reflux normal tissues (Normal) and patients with bile reflux gastritis (BRG). CD68 was utilized to label macrophage (green) and nuclei were stained with DAPI (blue). (E) mRNA levels of IL-1β and IL-18 in normal tissues or tissues from BRG patients were evaluated by qRT-PCR. GAPDH is used as an internal reference. (F, G) Levels of pyroptosis-related proteins in Normal and BRG patient tissues were detected by western blotting and quantified by Image J software. GAPDH served as a loading control. (H) Schematic diagram of gastrojejunostomy (GJ) surgery in mice. (I) Total BAs, Conjugated BAs, and TDCA were measured by LC-MS. (J) Gastric mucosal inflammation triggered by DGR was evaluated by H&E staining in mice. (K) Pyroptosis-related proteins in murine gastric tissues were assayed with western blotting between the Sham and GJ groups. (L, M) Different concentrations (stimulated for 24 h) and durations (100μM TDCA) of TDCA were added to U937 (L) or RAW264.7 cells (M) as a DRG model in vitro , followed by the detection of pyroptosis-related markers through western blotting. * P < 0.05, ** P < 0.01, and *** P <0.001.

Article Snippet: Taurodeoxycholic Acid (TDCA) (GC44995, GLPBIO, USA) was dissolved in DMSO and then added to the medium.

Techniques: Reflux, Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot, Software, Control, Liquid Chromatography with Mass Spectroscopy, In Vitro

Journal: Frontiers in Immunology

Article Title: USP50 regulates NLRP3 inflammasome activation in duodenogastric reflux-induced gastric tumorigenesis

doi: 10.3389/fimmu.2024.1326137

Figure Lengend Snippet: The expression of USP50 is upregulated in bile reflux gastritis and GC. (A) Protein levels of USP50 in gastric tissues from Normal, BRG, and GC (gastric cancer) patients were detected by western blotting. (B) Protein levels of USP50 in murine gastric tissues of the Sham and GJ groups were assayed with western blotting. (C) The expression of USP50 was detected by IF in normal tissues and BRG tissues (red). CD68 was used as a marker for macrophages(green), and the cell nuclei were stained with DAPI (blue). White arrows indicate co-localization of USP50 and CD68. (D) The number of USP50 positive cells per field was counted in the IF of gastric tissues (field: 20 X). (E) Percentage calculation of co-localization between USP50 and CD68. The co-localization percentage is determined by dividing the number of cells positive for both USP50 and CD68 by the number of cells positive for USP50 in the same randomly selected field. (field: 20 X) (F-I) U937 and RAW264.7 cells were subjected to TDCA stimulation at varying doses or durations. qRT-PCR was performed to detect the mRNA level of USP50 (F, G) , with western blotting used to assay the protein level of USP50 (H, I) . * P < 0.05, ** P < 0.01, and *** P <0.001.

Article Snippet: Taurodeoxycholic Acid (TDCA) (GC44995, GLPBIO, USA) was dissolved in DMSO and then added to the medium.

Techniques: Expressing, Reflux, Western Blot, Marker, Staining, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: USP50 regulates NLRP3 inflammasome activation in duodenogastric reflux-induced gastric tumorigenesis

doi: 10.3389/fimmu.2024.1326137

Figure Lengend Snippet: USP50 interacts with and deubiquitinates ASC to induce ASC speck formation and oligomerization. (A) Vector or Flag-USP50 was transferred to U937 cells and co-IP was performed with anti-Flag antibody. Whole-cell lysates (WCL) were used as Input. (B) ASC specks were detected by IF in U937 cells with or without USP50 knock-down. U937 cells were stimulated with TDCA (200μM) after being differentiated by PMA and primed by LPS. White arrowheads indicated ASC specks. (C) ASC specks of five random fields in each group were counted and analyzed with the student’s t-test. (* P <0.05). (D) Differentiated and primed U937 cells with or without USP50 overexpression were subjected to TDCA stimulation. DSS was used to crosslink ASC oligomers. Western blotting was utilized to analyze ASC aggregation. (E) U937 cells transfected with Vector or Flag-USP50 were lysed with IP lysis buffer and co-IP was conducted with anti-ASC antibody. Ub antibody was used to assay the ubiquitination of ASC by western blotting. MG132 (20μM) was added to the medium 6 hours before collecting cells. (F) WT, K48R, or K63R HA-Ub was co-transferred to 293T with His-ASC with or without Flag-USP50. co-IP was performed with His antibody and ubiquitination of ASC was analyzed with HA antibody via western blotting. MG132 was added to the culture medium 6 hours before collecting the cells.

Article Snippet: Taurodeoxycholic Acid (TDCA) (GC44995, GLPBIO, USA) was dissolved in DMSO and then added to the medium.

Techniques: Plasmid Preparation, Co-Immunoprecipitation Assay, Knockdown, Over Expression, Western Blot, Transfection, Lysis, Ubiquitin Proteomics

Journal: Frontiers in Immunology

Article Title: USP50 regulates NLRP3 inflammasome activation in duodenogastric reflux-induced gastric tumorigenesis

doi: 10.3389/fimmu.2024.1326137

Figure Lengend Snippet: USP50 stimulates HMGB1 release in DGR-induced gastric inflammation. (A, B) mRNA levels of HMGB1, HSP70, HSP90, S100A8, and S100A9 were detected via qRT-PCR in U937 cells (A) or RAW264.7 cells (B) treated with DMSO or TDCA (200μM, 24 h). (C-H) ELISA was used to measure the concentrations of HMGB1 (C) , HSP70 (D) , HSP90 (E) , S100A8 (F) , and S100A9 (G) in the supernatant of U937 cells and RAW264.7 cells, while ATP concentration (H) was surveyed by ATP assay kit. (I) The protein level of HMGB1 was detected in gastric tissues of Sham and GJ mice by western blotting. Image J was used for quantitative analysis, with GAPDH as loading control. (J) The level of HMGB1 between normal gastric tissues and gastric cancer tissues was analyzed via GEPIA database (gepia.cancer-pku.cn/). (K) The IHC staining of HMGB1 was performed with sections from BRG patients and GC patients. (L) IRS was calculated for (K) . (M, N) HMGB1 was assayed by ELISA in the supernatant of U937 cells with USP50 overexpression (M) or knock-down (N) . * P < 0.05, ** P < 0.01, and *** P <0.001. "ns" means "no significance" ( P >0.05).

Article Snippet: Taurodeoxycholic Acid (TDCA) (GC44995, GLPBIO, USA) was dissolved in DMSO and then added to the medium.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Concentration Assay, ATP Assay, Western Blot, Control, Immunohistochemistry, Over Expression, Knockdown

Journal: Frontiers in Immunology

Article Title: USP50 regulates NLRP3 inflammasome activation in duodenogastric reflux-induced gastric tumorigenesis

doi: 10.3389/fimmu.2024.1326137

Figure Lengend Snippet: USP50-HMGB1 axis contributes to DGR-induced gastric tumorigenesis. CM (conditioned media) was obtained from the supernatant of DMSO (DMSO-CM) or TDCA (TDCA-CM) stimulated parental U937 cells or USP50-modified U937 cells. Gastric cell lines SNU216 and HGC27 were cultured in CM. (A, B) CCK-8 assay was performed to assess cell viability of SNU216 (A) and HGC27 cells (B) in various groups including DMSO (DMSO-CM), TDCA (TDCA-CM), TDCA+HMGB1 NAs (TDCA-CM with HMGB1 NAs added), and rhHMGB1 (rhHMGB1 added to media). (C) Cell invasion ability in vitro was tested via Transwell assay. (D, E) Scratch assay was implemented to evaluate the cell motility of SNU216 (D) and HGC27 (E) . (F, G) The role of CM from USP50-overexpressed U937 cells and HMGB1 NAs on cell viability of SNU216 (F) and HGC27 (G) were assayed with a CCK-8 kit. (H, I) The effect of CM from USP50 knock-down U937 cells and rhHMGB1 on cell viability of SNU216 (H) and HGC27 (I) was detected by the CCK-8 method. (J, K) Transwell assay of SNU216 and HGC27 cells. Gastric cells were cultured with CM from USP50-overexpressed (J) or USP50-knockdown (K) U937 cells added with HMGB1 NAs or rhHMGB1. (L, M) Scratch assay was performed to evaluate cell motility. Ditto, SNU216 and HGC27 were cultured in CM, added with HMGB1 NAs or rhHMGB1. * P < 0.05, ** P < 0.01, and *** P <0.001.

Article Snippet: Taurodeoxycholic Acid (TDCA) (GC44995, GLPBIO, USA) was dissolved in DMSO and then added to the medium.

Techniques: Modification, Cell Culture, CCK-8 Assay, In Vitro, Transwell Assay, Wound Healing Assay, Knockdown

Journal: Frontiers in Immunology

Article Title: USP50 regulates NLRP3 inflammasome activation in duodenogastric reflux-induced gastric tumorigenesis

doi: 10.3389/fimmu.2024.1326137

Figure Lengend Snippet: USP50-HMGB1 axis promotes tumor proliferation and invasion through PI3K/AKT and MAPK/ERK pathways. CM was collected from the supernatant of USP50-modified U937 cells (USP50 or shUSP50) with or without TDCA stimulation (TDCA). CM was applied to gastric cells combined with or without HMGB1 NAs or rhHMGB1. (A) The effect of CM derived from U937 cells in diverse situations and HMGB1 NAs on key proteins of PI3K/AKT and MAPK/ERK pathways was detected in SNU216 and HGC27 cells via western blotting. (B) The effect of CM derived from TDCA-challenged, with or without USP50-knockdown U937 cells, combined with HMGB1 NAs on key proteins of PI3K/AKT and MAPK/ERK pathways was detected in SNU216 and HGC27 cells via western blotting. (C, D) CM combined with or without ERK1/2 inhibitor (SCH772984) and an AKT inhibitor (MK-2206) was used to culture SNU216 (C) and HGC27 (D) cells. Cell viability was evaluated with CCK-8 kits. (E-H) The effect of various types of CM, SCH772984, and MK-2206 on cell invasion (E, F) and cell mobility (G, H) in SNU216 and HGC27 cells. ** P < 0.01, and *** P <0.001.

Article Snippet: Taurodeoxycholic Acid (TDCA) (GC44995, GLPBIO, USA) was dissolved in DMSO and then added to the medium.

Techniques: Modification, Derivative Assay, Western Blot, Knockdown, CCK-8 Assay

Journal: iScience

Article Title: A physiologically based model of bile acid metabolism in mice

doi: 10.1016/j.isci.2023.107922

Figure Lengend Snippet: Physicochemical properties of bile acids used

Article Snippet: tDCA , pKa (acidic) , −0.75 , ChemAxon (HMDB, ) .

Techniques: Solubility