Journal: Bioengineered

Article Title: Male-specific long non-coding RNA testis-specific transcript, Y-linked 15 promotes gastric cancer cell growth by regulating Wnt family member 1/β-catenin signaling by sponging microRNA let-7a-5p

doi: 10.1080/21655979.2022.2053814

Figure Lengend Snippet: Effect of TTTY15 on GC cell proliferation, apoptosis, migration, invasion and EMT process. (a) The transfection efficiency of TTTY15 overexpression plasmid, si-TTTY15-1 and si-TTTY15-2 was detected by qRT-PCR. (b and c) CCK-8 method and BrdU assay were conducted for detecting the effects of TTTY15 overexpression and knockdown on the viability and proliferation of NCI-N87 and SUN-1 cells. (d) Flow cytometry assay was utilized for detecting the effects of TTTY15 overexpression and knockdown on the apoptosis of NCI-N87 and SUN-1 cells. (e) Transwell assays were performed to detect the effects of TTTY15 overexpression and knockdown on the migration and invasion of NCI-N87 and SUN-1 cells. (f) Western blot was carried out to detect the effects of TTTY15 overexpression and knockdown on the expression of EMT-related proteins in NCI-N87 and SUN-1 cells. All of the experiments were performed in triplicate. * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: The AnnexinV-FITC/PI cell apoptosis double staining kit (BD Biosciences, New Jersey, USA) was employed for detecting cell apoptosis.

Techniques: Migration, Transfection, Over Expression, Plasmid Preparation, Quantitative RT-PCR, CCK-8 Assay, BrdU Staining, Flow Cytometry, Western Blot, Expressing

Journal: Bioengineered

Article Title: Male-specific long non-coding RNA testis-specific transcript, Y-linked 15 promotes gastric cancer cell growth by regulating Wnt family member 1/β-catenin signaling by sponging microRNA let-7a-5p

doi: 10.1080/21655979.2022.2053814

Figure Lengend Snippet: Effects of TTTY15/let-7a-5p axis on GC cell proliferation, apoptosis, migration and invasion. (a) qRT-PCR was used to detect the transfection efficiency of TTTY15 overexpression plasmids + miR mimics and si-TTTY15-1 + miR inhibitors. (b and c) CCK-8 method and BrdU assay were used to detect the effects of TTTY15 and let-7a-5p on the viability and proliferation of NCI-N87 and SUN-1cells. (d) Flow cytometry assay was used to detect the effects of TTTY15 and let-7a-5p on NCI-N87 and SUN-1 cell apoptosis. (e and f) Transwell assays were used to detect the effects of TTTY15 and let-7a-5p on the migration and invasion of NCI-N87 and SUN-1 cells. (g) Western blot was used to detect the effects of TTTY15 and let-7a-5p on the EMT process of NCI-N87 and SUN-1 cells. All of the experiments were performed in triplicate. ** P < 0.01 and *** P < 0.001.

Article Snippet: The AnnexinV-FITC/PI cell apoptosis double staining kit (BD Biosciences, New Jersey, USA) was employed for detecting cell apoptosis.

Techniques: Migration, Quantitative RT-PCR, Transfection, Over Expression, CCK-8 Assay, BrdU Staining, Flow Cytometry, Western Blot

Journal: Acta Pharmaceutica Sinica. B

Article Title: Oridonin as a novel KDM5C inhibitor alleviates clonal hematopoiesis-induced cardiac aging via H3K4me3-dependent SASP suppression

doi: 10.1016/j.apsb.2026.02.006

Figure Lengend Snippet: Cellular co-culture models reveal that oridonin directly alleviates Tet2 +/− induced cardiac aging phenotypes in vitro . (A) Co-culture experimental design: PCs or PCFs were co-cultured with BMDMs from aged BMT mice, with or without oridonin treatment. (B) Representative images showing F4/80-labeled primary BMDMs from aged BMT mice displaying prominent filopodia. Scale bar, 20 μm. (C) Representative calcium recordings (left) of mouse PCs after co-culture with BMDMs. Right: quantification of calcium event frequency and calcium transient amplitude ( n = 30 cells). (D) Representative images (left) and quantification (right) of P21 (red) and phalloidin (green) staining in mouse PCs ( n = 30 cells). P21 expression levels were calculated as the mean fluorescence intensity using automated, unbiased quantification. Scale bar, 15 μm. (E) Representative images (left) and quantification (right) of SA- β- Gal staining in mouse PCs ( n = 100 cells from five independent experiments). Scale bar, 50 μm. (F) Representative images (left) and quantification (right) of lipofuscin deposition in mouse PCs, evaluated using the Schmorl iron reduction method ( n = 100 cells from four independent experiments). Scale bar, 50 μm. (G) Representative images (left) and quantification (right) of α -SMA (red) expression in mouse PCFs ( n = 30 cells). Scale bar, 50 μm. (H) Reactive oxygen species (ROS) generation in mouse PCs was analyzed by flow cytometry using DCFH-FA staining. Right: median ROS levels were quantified ( n = 5 samples per group). (I) Representative images (left) and quantification (right) of collagen lattice contraction by PCFs ( n = 3 samples per group). Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 by one-way ANOVA with Tukey's post hoc test (panels C–I). Abbreviations: WBM, whole bone marrow; BMDM, bone marrow-derived macrophages; PCs, primary cardiomyocytes; PCFs, primary cardiac fibroblasts; Ori, oridonin; BMT, bone marrow transplantation.

Article Snippet: Senescence-associated β -galactosidase (SA- β -Gal) staining was performed using the β -Galactosidase Staining Kit (Abbkine, Wuhan, China) according to the manufacturer's protocol.

Techniques: Co-Culture Assay, In Vitro, Cell Culture, Labeling, Staining, Expressing, Fluorescence, Flow Cytometry, Derivative Assay, Transplantation Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Oridonin as a novel KDM5C inhibitor alleviates clonal hematopoiesis-induced cardiac aging via H3K4me3-dependent SASP suppression

doi: 10.1016/j.apsb.2026.02.006

Figure Lengend Snippet: Oridonin reduces the rate of clonal hematopoiesis and cardiac aging induced by Tet2 +/ − through inhibition of KDM5C. (A) Experimental design. (B) Western blot analysis of KDM5C expression in BMDMs from Tet2 +/− BMT mice and after AAV-mediated KDM5C_OE. Longitudinal quantification of CD45.2-WT and CD45.2- Tet2 +/− leukocytes (C), monocytes (D), and macrophages (E) in myeloid cells of BMT mice. (F) Immunofluorescence staining of heart sections from BMT mice to detect the colocalization (yellow) of γ H2AX (red) and cTnT (green, cardiomyocyte-specific). Left: representative images (scale bar = 20 μm); Right: quantitative analysis of γ H2AX/cTnT colocalization. (G, H) Immunohistochemical analysis of S100A8 (G) and S100A9 (H) expression in BMT mouse hearts. Left: representative images (scale bar = 20 μm); Right: quantification of the proportion of positively stained cells. (I) Immunofluorescence staining for aggresome in BMT mouse hearts. Left: representative images (scale bar = 20 μm); Right: quantification of relative intensity. RT-qPCR analysis of Bnp (J) and Anp (K) expression in BMT mouse hearts. (L) Co-culture experimental design: BMDMs from BMT mice were co-cultured with PCs and treated with oridonin and/or AAV-mediated KDM5C_OE. (M) Representative images (left) and quantification (right) of SA- β- Gal staining in mouse PCs ( n = 100 cells from four independent experiments). (N) Mitochondrial membrane potential in PCs was analyzed by flow cytometry using JC-1 staining. Right: quantification of the proportion of JC-1 monomer cells ( n = 5 independent experiments). (O) Mitochondrial respiration analysis of mouse PCs using the Seahorse platform. (P–R) Quantification of basal oxygen consumption rate (OCR), maximal respiration, and ATP production. Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 by one-way ANOVA with Tukey's post hoc test ( n = 6 mice per group). Abbreviations: WBM, whole bone marrow; BMDM, bone marrow-derived macrophages; PCs, primary cardiomyocytes; PCFs, primary cardiac fibroblasts; Ori, oridonin; BMT, bone marrow transplantation. AAV, adeno-associated virus; KDM5C_OE, KDM5C overexpression; ns, not significant.

Article Snippet: Senescence-associated β -galactosidase (SA- β -Gal) staining was performed using the β -Galactosidase Staining Kit (Abbkine, Wuhan, China) according to the manufacturer's protocol.

Techniques: Inhibition, Western Blot, Expressing, Immunofluorescence, Staining, Immunohistochemical staining, Quantitative RT-PCR, Co-Culture Assay, Cell Culture, Membrane, Flow Cytometry, Derivative Assay, Transplantation Assay, Virus, Over Expression

Journal: Acta Pharmaceutica Sinica. B

Article Title: Oridonin as a novel KDM5C inhibitor alleviates clonal hematopoiesis-induced cardiac aging via H3K4me3-dependent SASP suppression

doi: 10.1016/j.apsb.2026.02.006

Figure Lengend Snippet: Validation of oridonin's effect in mitigating Tet2 mutation-driven CHIP cardiac aging via Sirt2 activation. (A) Differential expression of Sirt family members in the hearts of aged mice before and after oridonin treatment. GSEA plots showing DEGs associated with aging factors (B) and mitochondrial function (C) in GSE248894 treated with Sirt2 inhibitors. (D) Schematic diagram of the co-culture system. (E) RT-qPCR analysis showing the relative expression levels of S100a8 , S100a9 , Tnf , Il6 , and Ccl2 in BMDMs, Cdkn1a in primary cardiomyocytes (PCs) following co-culture treated with oridonin or Sirt2 inhibitors. (F) Representative images (left) and quantification (right) of P21 (red) and phalloidin (green) staining in mouse PCs ( n = 30 cells). P21 expression was quantified by mean fluorescence intensity using automated, unbiased analysis. Scale bar, 15 μm. (G) Representative images (left) and quantification (right) of SA- β- Gal staining in mouse PCs ( n = 100 cells from four independent experiments). Scale bar, 50 μm. (H) Representative images (left) and quantification (right) of lipofuscin deposition in mouse PCs, assessed by Schmorl iron reduction ( n = 100 cells from four independent experiments). Scale bar, 50 μm. (I) Representative images (left) and quantification (right) of α -SMA (red) expression in primary cardiac fibroblasts (PCFs; n = 30 cells). Scale bar, 50 μm. (J) Mitochondrial membrane potential in PCs analyzed via JC-1 staining and flow cytometry. Right: quantification of JC-1 monomer-positive cells ( n = 4 independent experiments). (K) Mitochondrial respiration analysis of PCs using the Seahorse platform. (L–N) Quantification of basal oxygen consumption rate (OCR), maximal respiration, and ATP production. Data are presented as mean ± SEM. Statistical analysis by one-way ANOVA with Tukey's post hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Abbreviations: WBM, whole bone marrow; BMDM, bone marrow-derived macrophages; PCs, primary cardiomyocytes; PCFs, primary cardiac fibroblasts; Ori, oridonin; BMT, bone marrow transplantation; ns, not significant.

Article Snippet: Senescence-associated β -galactosidase (SA- β -Gal) staining was performed using the β -Galactosidase Staining Kit (Abbkine, Wuhan, China) according to the manufacturer's protocol.

Techniques: Biomarker Discovery, Mutagenesis, Activation Assay, Quantitative Proteomics, Co-Culture Assay, Quantitative RT-PCR, Expressing, Staining, Fluorescence, Membrane, Flow Cytometry, Derivative Assay, Transplantation Assay