Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: MDGA2 is inactivated by promoter methylation in gastric cancer cell lines. (A) MDGA2 was silenced or downregulated in 10 out of 11 gastric cancer cell lines but readily expression in normal gastric tissue. The methylation status of MDGA2 was determined by methylation-specific PCR (MSP). M, methylated; U, unmethylated. (B) A typical CpG island is present at the promoter region of MDGA2. Each vertical bar represents a single CpG site. The transcription start site (TSS) is indicated by a curved arrow. A region for bisulfite genomic sequencing (BGS) and combined bisulfite restriction analysis (COBRA) is denoted. BGS analysis confirmed high levels of promoter methylation in MDGA2-silenced cells and no/low methylation in MDGA2-expressing samples. (C) MDGA2 mRNA expression was restored after treatment with the demethylation reagent 5-Aza. Decreased methylation was revealed by BGS after 5-Aza treatment.

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: Methylation, Expressing, Genomic Sequencing, Combined Bisulfite Restriction Analysis Assay

Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: Epigenetic inactivation of MDGA2 in primary gastric cancers. (A) Transcriptional downregulation of MDGA2 in gastric tumours compared with adjacent normal tissues was shown by reverse transcription PCR (RT-PCR). (B) Protein levels of MDGA2 were quantitated as the percentage of cells with positive staining by immunohistochemistry in 20 pairs of gastric tumour and adjacent non-tumour samples. (C) Methylation-specific PCR (MSP) showed high methylation of MDGA2 promoter in 10 gastric tumours and low methylation in their adjacent non-tumour tissues, which was confirmed by bisulfite genomic sequencing (BGS). M, methylated; U, unmethylated. (D) Kaplan–Meier curves showed that MDGA2 methylation is significantly associated with shortened survival in overall patients with gastric cancer and (E) in patients at an early stage but not patients at late stages (log rank test).

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Staining, Immunohistochemistry, Methylation, Genomic Sequencing

Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: Univariate and multivariate Cox regression analyses of potential poor prognostic factors in gastric cancer

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: Methylation

Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: In vitro gain- and loss-of-function assays on MDGA2. (A) Ectopic expression of MDGA2 in AGS and BGC823 cells at mRNA and protein levels was confirmed by reverse transcription PCR (RT-PCR) and western blot analysis. (B) MDGA2 significantly inhibited cell viability and colony formation ability. (C) MDGA2 caused cell cycle arrest at G1–S transition, as indicated by flow cytometry. Reduced cell proliferation by MDGA2 was further shown by Ki-67 staining (magnification ×400) and altered cell cycle-related protein expression by western blot analysis. (D) Cell apoptosis by flow cytometry analysis after annexin V-APC and 7-aminoactinomycin (7-AAD) double staining. Q2 shows the late apoptotic cells and Q4 shows the early apoptotic cells. Cell apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) staining (magnification ×400) and upregulation of apoptosis-related proteins in MDGA2-expressing cells was confirmed by western blot analysis. (E) Growth inhibitory effect of cisplatin on MDGA2-overexpressing and control vector-transfected AGS and BGC823 cells. Cell viability was measured by MTT assay after 48 h treatment with cisplatin. Data are mean±SD from three independent experiments. (F) Knockdown of MDGA2 in MKN1 cells by short interference RNA (siRNA) transfection was confirmed by RT-PCR, qRT-PCR and western blot analysis. (G) Knockdown of MDGA2 significantly increased cell viability of MKN1 cells, promoted colony formation and (H) promoted cell cycle progression, but reduced cell apoptosis as indicated by flow cytometry. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: In Vitro, Expressing, Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Staining, Double Staining, End Labeling, TUNEL Assay, Control, Plasmid Preparation, Transfection, MTT Assay, Knockdown, Quantitative RT-PCR

Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: In vivo subcutaneous and orthotopic xenograft models confirmed the suppressive effect of MDGA2 on tumorigenicity. (A) Subcutaneous tumour growth curve of MDGA2-expressing BGC823 cells in nude mice was compared with control vector (pcDNA3.1) transfected cells. The data are mean±SD (n=5/group) of three separate experiments. (B) A representative image of tumour growth in nude mice subcutaneously inoculated with MDGA2- or control vector-transfected cells. Tumour weight was compared at the end of the experiment. MDGA2 expression in subcutaneous xenografts was confirmed by immunohistochemistry (IHC). (C) Cell proliferative activity was evaluated by Ki-67 staining and apoptotic activity by terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) staining in subcutaneous xenografts. (D) Representative images of orthotopic xenograft tumours. Both volume and weight of the orthotopic xenograft tumours were significantly smaller in the MDGA2 group than in the control group. (E) Confirmation of MDGA2 expression in orthotopic xenograft tumours by IHC. (F) Ki-67 staining in orthotopic xenografts and TUNEL staining in orthotopic xenografts. *p<0.05, **p<0.001, ***p<0.0001.

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: In Vivo, Expressing, Control, Plasmid Preparation, Transfection, Immunohistochemistry, Activity Assay, Staining, End Labeling, TUNEL Assay

Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: MDGA2 interacts with DNA methyltransferase 1 associated protein 1 (DMAP1) in gastric cancer. (A) Co-immunoprecipitation (Co-IP) of MDGA2 binding proteins followed by mass spectrometry identified DMAP1 to be a MDGA2-binding protein. (B) Co-IP of MDGA2 and DMAP1 with each other from proteins of MDGA2-transfected AGS and BGC823 cells. The presence of MDGA2 and DMAP1 in the Co-IP products was confirmed by western blot analysis using specific antibodies. (C) Direct interaction between MDGA2 and DMAP1 was shown by glutathione S-transferase (GST) pull-down assay. (D) Western blot analysis for DMAP1 expression. (E) MDGA2 increased the DMAP1 level by inhibiting its ubiquitin-mediated degradation. (F1) Confocal immunofluorescence analysis of MDGA2 and DMAP1 expression in MDGA2- and empty vector-transfected cells. The scale bar of 50 M is for unframed images of both cell groups. The yellow-frame fields were enlarged arbitrarily. (F2) Western blot detection of MDGA2 and DMAP1 in the membrane, cytoplasmic and nuclear fractions of AGS and BGC823 cells transfected with MDGA2 expression vector. (G) Western blot analysis showed increased levels of ataxia telangiectasia mutated (ATM) and p53 by MDGA2 over-expression in AGS and BGC823 cells and xenograft tumours.

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Binding Assay, Mass Spectrometry, Transfection, Western Blot, Pull Down Assay, Expressing, Ubiquitin Proteomics, Immunofluorescence, Plasmid Preparation, Membrane, Over Expression

Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: Suppression of gastric cancer growth by MDGA2 is partly dependent on DNA methyltransferase 1 associated protein 1 (DMAP1). (A) Expression of MDGA2 was not affected by DMAP1. (B) Cell viability of AGS and BGC823 cells by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. (C) Colony formation in AGS and BGC823 cells. (D) DMAP1 was knocked down in cells with stable MDGA2 over-expression by short interference RNA (siRNA) transfection. (E) Effect on gastric cancer cell growth by different levels of MDGA2 and DMAP1. Cell growth was monitored by the xCelligence system. Data shown are mean±SD. (F) Effect of different levels of MDGA2 and DMAP1 on colony formation ability of gastric cancer cells. *p<0.05, **p<0.001, ***p<0.0001.

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: Expressing, MTS Assay, Over Expression, Transfection

Journal: Gut

Article Title: MDGA2 is a novel tumour suppressor cooperating with DMAP1 in gastric cancer and is associated with disease outcome

doi: 10.1136/gutjnl-2015-309276

Figure Lengend Snippet: MDGA2 induces p53/p21 signalling pathway. (A) Luciferase reporter assays showed that p53 and p21 were significantly activated by MDGA2 expression. Date are mean±SD. *p<0.05, **p<0.001. (B) p53 knockdown by short interference RNA (siRNA) in cells with stable MDGA2 transfection was examined by western blot analysis. (C) p53 knockdown in AGS and BGC823 cells transfected with MDGA2 partially blunts the effects of MDGA2 overexpression on cell growth, as evidenced by cell viability and (D) colony formation assays. Data shown are mean±SD. *p<0.05, **p<0.01, ***p<0.005. (E) Downstream targets of MDGA2 identified by p53 signalling pathway PCR array. (F) Schematic illustration of the molecular mechanism of MDGA2 in gastric cancer. MDGA2 binds to DNA methyltransferase 1 associated protein 1 (DMAP1) to stabilise DMAP1 by inhibiting its ubiquitin-mediated degradation, which subsequently activates the ataxia telangiectasia mutated (ATM)/p53/p21 signalling pathway to mediate the tumour suppressive function by inhibiting cell cycle progression and promoting apoptosis. Besides activation of p21 and p53, upregulation of p21 ( C), ATM and p53 ( G) by MDGA2 was well confirmed at the protein level by western blot analysis.

Article Snippet: Purified recombinant human glutathione S-transferase (GST)-DMAP1 protein (Abnova, Taipei City, Taiwan) and His-tagged recombinant human MDGA2 (MDGA2-His; Creative Biomart, Shirley, New York, USA) were used for pull-down assay.

Techniques: Luciferase, Expressing, Knockdown, Transfection, Western Blot, Over Expression, Ubiquitin Proteomics, Activation Assay

Journal: Orthopaedic Journal of Sports Medicine

Article Title: Morphological Changes in Femoral Trochlea After Extensor Realignment Surgery in Children With Congenital Patellar Dislocation

doi: 10.1177/23259671261418048

Figure Lengend Snippet: Surgical procedure. (A) Patella fixed on the lateral side of the lateral condyle of the distal femur. (B) Lateral retinacular release. (C) Resection of the redundant medial capsule. (D) Patella substantially returned to the central position of the knee joint. (E) Patella sutured to the remaining medial capsule. (F) Lateral half of the patellar ligament separated and woven. (G) Lateral half of the patellar ligament transposed medially. (H) Absorbable screw fixation.

Article Snippet: An absorbable suture anchor (2.9 mm; Johnson & Johnson) was used to fix the tendon to the anteromedial tibial metaphysis ( ).

Techniques:

Journal: Arthroscopy Techniques

Article Title: Capsulolabral Advancement and “Suture-First” Knot-Less Technique for Chronic Posterior Glenolabral Articular Disruption Lesion of the Shoulder

doi: 10.1016/j.eats.2025.103564

Figure Lengend Snippet: Arthroscopic view of right shoulder from the antero-superior viewing portal with patient in lateral position showing both ends of the SutureTape being loaded on a 2.9-mm PEEK knotless suture anchor (PushLock, Arthrex), and introduced through the posterior working portal.

Article Snippet: A second SutureTape is passed through proximal part of detached labrum and adjacent capsule and both ends of SutureTape loaded to a second PushLock anchor (2.9-mm PEEK; Arthrex), which is fixed to the proximal part of margin of defect ( , ).

Techniques:

Journal: Arthroscopy Techniques

Article Title: Capsulolabral Advancement and “Suture-First” Knot-Less Technique for Chronic Posterior Glenolabral Articular Disruption Lesion of the Shoulder

doi: 10.1016/j.eats.2025.103564

Figure Lengend Snippet: Now visualising from the antero-superior portal, after appropriate size drilling of the pilot hole using the posterior portal, the PushLock anchor with the SutureTape is fixed on the margin of the defect.

Article Snippet: A second SutureTape is passed through proximal part of detached labrum and adjacent capsule and both ends of SutureTape loaded to a second PushLock anchor (2.9-mm PEEK; Arthrex), which is fixed to the proximal part of margin of defect ( , ).

Techniques:

Journal: Arthroscopy Techniques

Article Title: Capsulolabral Advancement and “Suture-First” Knot-Less Technique for Chronic Posterior Glenolabral Articular Disruption Lesion of the Shoulder

doi: 10.1016/j.eats.2025.103564

Figure Lengend Snippet: Now, using the same viewing portal and working portal, on right shoulder of the patient in lateral decubitus position, a second SutureTape and second PushLock anchor are inserted to fix proximal part of margin of defect.

Article Snippet: A second SutureTape is passed through proximal part of detached labrum and adjacent capsule and both ends of SutureTape loaded to a second PushLock anchor (2.9-mm PEEK; Arthrex), which is fixed to the proximal part of margin of defect ( , ).

Techniques: