Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 2 Identification of PKM2 as a direct binding target for PCA. (A) The employed drug affinity responsive target stabilization (DARTS) assay detects a marked increase in w60 kD band upon PCA incubation in pronase digested H9C2 cell lysates. (B) Immunoblot analysis of PKM2 in pronase-digested cell lysate (n Z 3). (C) Recombinant PKM2 in pronase-digested H9C2 cell lysates. PKM2 degradation in H9C2 cell lysates (D) and in intact cells (E) (n Z 3). (F) Surface plasmon resonance (SPR) binding curve fit to a Kd Z 5.76 nmol/L for PCA and PKM2 (n Z 3). Results are shown as mean SD; *P < 0.05, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Binding Assay, Incubation, Western Blot, Recombinant, SPR Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 3 PCA PKM2-dependently protects cardiomyocytes. Primary neonatal rat ventricular myocytes (NRVMs) were cultured in glucose-free DMEM under 1% O2 (OGD) for 4 h in the presence of protocatechuic aldehyde (PCA). (A) Cell survival in NRVMs exposed to OGD for 6 h (n Z 6). (B) Intracellular ROS production (n Z 6). (C) Representative images of mitochondrial permeability transition pore (mPTP) and quantification of relative fluorescence intensity (n Z 6, one of three independent experiments). Scale bar: 10 mm. (D) Mitochondrial fission was detected by Mito-Tracker Red (one of 3 independent experiments) and quantification analysis. Scale bar: 10 mm. (E) Caspase 3 activity in NRVMs (n Z 6). (F) Representative Western blots of BAX, BCL-2, caspase 3, cleaved caspase 3 (c-caspase 3) and the ratio of BAX/BCL-2 (n Z 3). (G) Representative images of TUNEL staining and quantification analysis the percentage of apoptosis cells. Scale bars, 100 mm (n Z 3). (H) Data of Annexin V/PI double staining flow cytometry; the percentage for each panel indicates the percentage of apoptotic cells and quantification of the percentage of apoptotic cells (n Z 3). Results are shown as mean SD; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Cell Culture, Permeability, Activity Assay, Western Blot, TUNEL Assay, Staining, Double Staining, Cytometry

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 4 PCA promotes the nuclear localization of PKM2. Primary neonatal rat ventricular myocytes (NRVMs) were exposed to 1% O2 in glucose-free DMEM (OGD) for 4 h in the presence of protocatechuic aldehyde (PCA). (A) PKM2 activity in NRVMs (n Z 6). (B) Immuno- fluorescence image of endogenous PKM2 (one of three independent experiments. Scale bar: 10 mm. (C) Nuclear PKM2 protein expression (n Z 3). (D) Cytoplasmic PKM2 protein expression (n Z 3). (E) Total PKM2 protein expression (n Z 3). (F) Docking analysis illustrates the interaction between PCA and PKM2. The residues that are likely to participate in the interactions with PCA are labeled. (G) Representative co- immunoprecipitation (co-IP) analysis of PKM2 and SIRT6 in the NRVMs (n Z 3). (H) Representative co-IP of PKM2 and b-catenin in the NRVMs (n Z 3). Results are shown as mean SD; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Activity Assay, Expressing, Labeling, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 6 The myocardial protection effect of PCA is dependent on PKM2. Mice were orally administrated with protocatechuic aldehyde (PCA) for 3 weeks after coronary artery ligation. (A) Representative photomicrographs for HE staining of cardiac tissue sections (n Z 6), Scale bar, 50 mm. (B) Representative photomicrographs for masson staining of cardiac tissue sections (n Z 6). Scale bar, 50 mm. (C) 4-HNE contents in the heart (n Z 6). (D) Representative M mode images of echocardiography from the assayed groups under treatments as indicated (n Z 6). (E) Ejection fractions (EF) and shortening fraction (FS) in mice (n Z 6). (F) Representative co-IP analysis of PKM2 and b-catenin in the heart (n Z 3). The mRNA levels of Myc (G), Ccnd1 (H) and Sgk1 (I) in the heart (n Z 6). (J) Immunocytochemical staining of BAX, BCL-2 and c- caspase 3 protein expression (n Z 6), scale bar Z 50 mm. (K) Representative TUNEL staining images and quantification of TUNEL positive cells (n Z 6). Results are shown as mean SD; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies against GAPDH (60004-1-Ig), b-catenin (51067-2-AP, 66379-1-Ig), PCNA (10205-2-AP), caspase 3 (66470-2-Ig), BAX (60267-1-Ig), BCL-2 (60178-1-Ig), PKM2 (60268-1-Ig), b-tubulin (10068-1-AP) and TCF4 (22337-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Ligation, Staining, Co-Immunoprecipitation Assay, Expressing, TUNEL Assay

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Transcriptomics-Based Liquid Biopsy for Early Detection of Recurrence in Locally Advanced Gastric Cancer.

doi: 10.1002/advs.202406276

Figure Lengend Snippet: Figure 2. Discovery process and preliminary validation of candidate markers for postoperative recurrence in LAGC patients based on public databases and transcriptomics sequencing data. A) Four candidate mRNAs (AGTR1, DNER, EPHA7, SUSD5) were identified through a Venn diagram analysis using the TCGA database (28 recurrent patients versus 159 non-recurrent patients), transcriptome data from the GEO database (125 recurrent patients versus 157 non-recurrent patients), and paired mRNA sequencing (3 recurrent patients versus 3 non-recurrent patients). B) A volcano plot illustrates the expression levels of these four genes in recurrent and non-recurrent cancer tissues. C) The expression levels of the four candidate mRNAs (AGTR1,

Article Snippet: Membranes were probed with antibodies against AGTR1 (1:1000 dilutions, 25343-1-AP, Proteintech, Chicago, USA), DNER (1:1000 dilutions, 24362-1-AP, Proteintech, Chicago, USA), EPHA7 (1:1000 dilutions, 66667-1-Ig, Proteintech, Chicago, USA), SUSD5 (1:500 dilutions, bs-7331R, Bioss, Beijing, China), CREB (1:500 dilutions, 381013, Zenbio, Chengdu, China), p-CREB (1:500 dilutions, 380697, Zenbio, Chengdu, China), and GAPDH (1:10000 dilutions, 10494-1-AP, Proteintech, Chicago, USA).

Techniques: Biomarker Discovery, Sequencing, Expressing

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Transcriptomics-Based Liquid Biopsy for Early Detection of Recurrence in Locally Advanced Gastric Cancer.

doi: 10.1002/advs.202406276

Figure Lengend Snippet: Figure 7. Four recurrence-related mRNA genes promote GC cell proliferation, migration and invasion in vitro. A,B) Scratch assay to evaluate the migration ability of GC cells after knockdown of AGTR1 and DNER, respectively. C,D) Transwell assay to assess the invasion and metastasis abilities of GC cells after knockdown of AGTR1 and DNER, respectively. E,F) EdU assay to determine the proliferation ability of GC cells after knockdown of AGTR1 and DNER, respectively. G–J) Colony formation assay to measure the proliferation ability of GC cells after knockdown of AGTR1, DNER, EPHA7, and SUSD5. K–N) CCK-8 assay to detect the proliferation ability of GC cells after knockdown of AGTR1, DNER, EPHA7, and SUSD5. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Membranes were probed with antibodies against AGTR1 (1:1000 dilutions, 25343-1-AP, Proteintech, Chicago, USA), DNER (1:1000 dilutions, 24362-1-AP, Proteintech, Chicago, USA), EPHA7 (1:1000 dilutions, 66667-1-Ig, Proteintech, Chicago, USA), SUSD5 (1:500 dilutions, bs-7331R, Bioss, Beijing, China), CREB (1:500 dilutions, 381013, Zenbio, Chengdu, China), p-CREB (1:500 dilutions, 380697, Zenbio, Chengdu, China), and GAPDH (1:10000 dilutions, 10494-1-AP, Proteintech, Chicago, USA).

Techniques: Migration, In Vitro, Wound Healing Assay, Knockdown, Transwell Assay, EdU Assay, Colony Assay, CCK-8 Assay

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Transcriptomics-Based Liquid Biopsy for Early Detection of Recurrence in Locally Advanced Gastric Cancer.

doi: 10.1002/advs.202406276

Figure Lengend Snippet: Figure 8. Four recurrence-related mRNA genes promote GC cell xenograft tumor growth and metastasis in vivo. A–D) Morphological images showing reduced subcutaneous xenograft tumor formation in mice injected with AGS cells knocked down for AGTR1, DNER, EPHA7, and SUSD5, along with tumor volume growth curves and final tumor weights. E) Representative IHC images of subcutaneous xenograft tumors after knockdown of AGTR1 (left), and quantification of IHC staining data for Ki67, N-cadherin, E-cadherin, and Vimentin in each group of mice (right). F) Representative IHC images of peritoneal metastasis tumors after intraperitoneal injection of AGS cells knocked down for AGTR1 (left), and quantification of IHC staining data for MMP9, N-cadherin, E-cadherin, and Vimentin in each group of mice (right). G) Representative images of peritoneal metastasis tumors in the abdominal cavity of mice injected with AGS cells knocked down for AGTR1. H) Measurement and quantification of the number of peritoneal metastatic tumors in

Article Snippet: Membranes were probed with antibodies against AGTR1 (1:1000 dilutions, 25343-1-AP, Proteintech, Chicago, USA), DNER (1:1000 dilutions, 24362-1-AP, Proteintech, Chicago, USA), EPHA7 (1:1000 dilutions, 66667-1-Ig, Proteintech, Chicago, USA), SUSD5 (1:500 dilutions, bs-7331R, Bioss, Beijing, China), CREB (1:500 dilutions, 381013, Zenbio, Chengdu, China), p-CREB (1:500 dilutions, 380697, Zenbio, Chengdu, China), and GAPDH (1:10000 dilutions, 10494-1-AP, Proteintech, Chicago, USA).

Techniques: In Vivo, Injection, Knockdown, Immunohistochemistry

Journal: Hereditas

Article Title: Corynoxine suppresses lung adenocarcinoma proliferation and metastasis via inhibiting PI3K/AKT pathway and suppressing Cyclooxygenase-2 expression.

doi: 10.1186/s41065-024-00343-x

Figure Lengend Snippet: Fig. 4 Corynoxine reduces COX-2 levels through PI3K/ATK pathway suppression. After treated with different concentrations of Corynoxine, (a) COX-2 mRNA expression was detected by qRT-PCR. (b) Western immunoblotting was used to detect COX-2 and β-actin levels. (c) After treated with different concentrations of Corynoxine or Celecoxib (40 µM) for 24 h, the level of PEG2 was evaluated by ELISA assay. (d) A549 cells were treated with indicated doses of Corynoxine for 24 h after pretreatment with the COX-2 selective inhibitor Celecoxib (40 µM) for 8 h, and the cell viability was determined by CCK-8 assay. After treated with different concentrations of Corynoxine. (e) Different subunits of PI3K (PIK3CA, PIK3CB and PIK3CD) were detected by qRT- PCR. (f) Western immunoblotting was used to detect PI3K p110δ, p-AKT, AKT, and β-actin levels. (g) After treatment with 100 µM Corynoxine or 10 µM LY294002 for 24 h, western immunoblotting was used to assess the expression of COX-2 and β-actin expression. Data were expressed as mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Blots were incubated overnight with primary antibodies prepared in 5% BSA (Solarbio, SW3015, China) at 4 °C, including antibodies specific for Vimentin (1:1000, Abcam, ab20346, USA), Bcl-2 (1:2000, Proteintech, 26593-1-AP, China), Bax (1:200, Santa Cruz, sc-20067, USA), E-cadherin (1:1000, CST, 14472, USA), PI3K p110δ (1:200, Santa Cruz, sc-55589, USA), AKT (1:1000, CST, 4685, USA), p-AKT (1:1000, CST, 4060, USA), and COX-2 (1:1000, CST, 12282, USA), and β-actin (1:5000, TransGen, TC201, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, CCK-8 Assay

Journal: Hereditas

Article Title: Corynoxine suppresses lung adenocarcinoma proliferation and metastasis via inhibiting PI3K/AKT pathway and suppressing Cyclooxygenase-2 expression.

doi: 10.1186/s41065-024-00343-x

Figure Lengend Snippet: Fig. 6 Corynoxine suppresses intratumoral PI3K/AKT/COX-2 pathway activity. (a) Bax, Bcl-2, and β-actin protein levels. (b) E-cadherin, Vimentin, and β-actin protein levels. (c) Intratumoral COX-2, PI3K, AKT, p-AKT, and β-actin levels were detected. Data were expressed as mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Blots were incubated overnight with primary antibodies prepared in 5% BSA (Solarbio, SW3015, China) at 4 °C, including antibodies specific for Vimentin (1:1000, Abcam, ab20346, USA), Bcl-2 (1:2000, Proteintech, 26593-1-AP, China), Bax (1:200, Santa Cruz, sc-20067, USA), E-cadherin (1:1000, CST, 14472, USA), PI3K p110δ (1:200, Santa Cruz, sc-55589, USA), AKT (1:1000, CST, 4685, USA), p-AKT (1:1000, CST, 4060, USA), and COX-2 (1:1000, CST, 12282, USA), and β-actin (1:5000, TransGen, TC201, China).

Techniques: Activity Assay