Journal: Hepatology Communications

Article Title: Proteomics‐based identification of the role of osteosarcoma amplified‐9 in hepatocellular carcinoma recurrence

doi: 10.1002/hep4.1952

Figure Lengend Snippet: Differential proteins expression in OS‐9 overexpression group (n = 3) and vector control (n = 3) of SMMC‐7721 cells. (A) Volcano plot of the differentially expressed proteins. Gray dots represent genes that are not differentially expressed in the early recurrence group and nonrecurrence group; red dots and blue dots represent genes that are up‐regulated and down‐regulated significantly in the early recurrence group. (B) Heat map of the differentially expressed proteins. Red rectangles mean that genes are up‐regulated in these samples, and blue ones mean down‐regulated. Two hundred and sixty‐eight protein expressions were up‐regulated and 140 protein expressions were down‐regulated in the overexpression group compared with the vector control group (fold change ≥ 1.5; p < 0.05). (C) Heat map of the differentially expressed proteins classified to the hypoxia‐inducible factor 1 (HIF‐1) and tumor necrosis factor (TNF) signaling pathway. (D) Ridgeline plot of Kyoto Encyclopedia of Genes and Genomes pathway enrichment for differentially expressed proteins. (E) Gene Ontology functions for differentially expressed proteins. The left side of the circle includes all related genes, and the right side displays the Gene Ontology terms. Red and blue rectangles mean that genes are up‐regulated and down‐regulated in the early recurrence group. Abbreviations: ALDOA, aldolase, fructose‐bisphosphate A; ALDOC, aldolase, fructose‐bisphosphate C; BCL10, BCL10 immune signaling adaptor; CASP7, caspase 7; CCNB1, cyclin B1; CDK6, cyclin dependent kinase 6; CEBPB, CCAAT enhancer binding protein beta; CHEK2, checkpoint kinase 2; CREBBP, CREB binding protein; DDX58, DExD/H‐box helicase 58; ENO1, enolase 1; ENO2, enolase 2; ENO3, enolase 3; HK2, hexokinase 2; HMOX1, heme oxygenase 1; IGFBP3, insulin like growth factor binding protein 3; JUNB, JunB proto‐oncogene; LDHA, lactate dehydrogenase A; MALT1, MALT1 paracaspase; MLKL, mixed lineage kinase domain like pseudokinase; OE, overexpressed; PGK1, phosphoglycerate kinase 1; PLCG2, phospholipase C gamma 2; SERPINE1, serpin family E member 1; SFN, stratifin. NC, control; STEAP3, STEAP3 metalloreductase; TNFAIP3, TNF alpha induced protein 3; TRAF5, TNF receptor associated factor 5

Article Snippet: Inhibitors including HIF‐1α and tumor necrosis factor alpha (TNF‐α) were obtained (VH‐298, MedChemExpress; HY‐100947, Methylthiouracil; HY‐B0513, MedChemExpress).

Techniques: Expressing, Over Expression, Plasmid Preparation, Control, Binding Assay

Journal: Hepatology Communications

Article Title: Proteomics‐based identification of the role of osteosarcoma amplified‐9 in hepatocellular carcinoma recurrence

doi: 10.1002/hep4.1952

Figure Lengend Snippet: (A–K) The relative messenger RNA (mRNA) expression level of lineage kinase domain‐like MLKL (A), tumor necrosis factor alpha–induced protein 3 (TNFAIP3) (B), JunB proto‐oncogene (JUNB) (C), and tumor necrosis factor receptor–associated factor 5 (TRAF5) (D), TNF‐α (E) and caspase‐7 (CASP7) (F) could be observed to increase more than 2‐fold ( p < 0.01). The relative expression of enolase1 (ENO1) (G), enolase2 (ENO2) (H), enolase3 (ENO3) (I), aldolase, fructose‐bisphosphate A (ALDOA) (J), and lactate dehydrogenase A (LDHA) (K) were also elevated more than 2‐fold ( p < 0.01)

Article Snippet: Inhibitors including HIF‐1α and tumor necrosis factor alpha (TNF‐α) were obtained (VH‐298, MedChemExpress; HY‐100947, Methylthiouracil; HY‐B0513, MedChemExpress).

Techniques: Expressing

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Reduced Plasma Kallistatin Is Associated With the Severity of Coronary Artery Disease, and Kallistatin Treatment Attenuates Atherosclerotic Plaque Formation in Mice

doi: 10.1161/JAHA.118.009562

Figure Lengend Snippet: Kallistatin inhibited low‐shear stress–induced carotid artery plaque formation. A , Representative magnetic resonance images in cross‐sectional views through the carotid arteries. The red arrow indicates the cross section of the left carotid artery, and the blue arrow indicates the cross section of the right carotid artery. B , Quantitative analysis of left carotid artery diameter in the Ad. HKS and Ad.Null groups. L‐ NAME and NAM blocked the effects of kallistatin (n=10, * P <0.05 vs the Ad.Null‐, L‐ NAME ‐, or NAM ‐treated groups). C , Quantitative analysis of left carotid artery plaque volume in the Ad. HKS and Ad.Null groups. L‐ NAME and NAM blocked the effect of kallistatin; n=10 (* P <0.05 vs the Ad.Null‐, L‐ NAME ‐ or NAM ‐treated groups). D , Plasma MDA levels in apoE −/− mice. E , Plasma TNF ‐α levels in apoE −/− mice. F , Distribution of mouse kallistatin expression in atherosclerotic lesions and normal vascular tissue. Data are presented as the mean± SEM ; n=10, * P <0.05 vs the Ad.Null‐, L‐ NAME ‐, or NAM ‐treated groups. Ad.HKS indicates adenovirus containing kallistatin cDNA; Ad.Null, adenovirus containing null cDNA; L‐NAME, N ω ‐nitro‐L‐arginine methyl ester; MDA, malondialdehyde; NAM, nicotinamide; SEM, standard error of the mean; TNF‐α, tumor necrosis factor‐α.

Article Snippet: Plasma samples were used for the analysis of TNF‐α using a Mouse TNF‐α ELISA (Proteintech, Rosemount, IL) according to the manufacturer's protocol.

Techniques: Shear, Clinical Proteomics, Expressing

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Reduced Plasma Kallistatin Is Associated With the Severity of Coronary Artery Disease, and Kallistatin Treatment Attenuates Atherosclerotic Plaque Formation in Mice

doi: 10.1161/JAHA.118.009562

Figure Lengend Snippet: Kallistatin led to morphological changes in the atherosclerotic plaques of apoE −/− mice. A , Representative images of HE staining. Original magnification is ×10 (n=5 in each group). B , Oil red O staining of carotid specimens of mice in the Ad.Null group, the Ad. HKS group, and L‐ NAME ‐ or NAM ‐treated groups (n=5 in each group). C , Representative images of superoxide formation labeled by the red fluorescence dye dihydroethidium in the carotid artery of Ad.Null mice, Ad. HKS mice, Ad. HKS +L‐ NAME mice, and Ad. HKS + NAM mice (n=5 in each group). D , Representative images of CD 68 immunostaining in lesion areas of the left carotid artery (n=5 in each group). E , Immunohistochemical assessments of the protein level of TNF ‐α in carotid plaque (n=5 in each group). F, Quantitative analyses of CD 68‐positive cells in the 4 groups (n=5 in each group, * P <0.05 vs the Ad.Null‐, L‐ NAME ‐, or NAM ‐treated groups). G , Fluorescence intensity was measured by a fluorescence microscope and quantified with Image software (n=5 in each group, * P <0.05 vs the Ad.Null‐, L‐ NAME ‐, or NAM ‐treated groups). H , Colocalization of PE red fluorescence ( SIRT 1) and FITC green fluorescence ( CD 31) within plaques (×40 magnification) in the carotid arteries of the Ad.Null, Ad. HKS , Ad. HKS +L‐ NAME , and Ad. HKS + NAM groups with labeled cell nuclei (blue) (n=5 in each group). Regions shown at a higher magnification (far right) are indicated by yellow boxes. Endothelial cell‐specific SIRT 1 fluorescence was measured on the luminal side of the internal elastic lamina only and expressed in mean pixel intensity per area (n=5 in each group, # P <0.05 vs the Ad.Null‐ and NAM ‐treated groups). I , Colocalization of PE red fluorescence ( eNOS ) and FITC green fluorescence ( CD 31) within carotid artery plaques (×40 magnification) of the 4 groups with labeled cell nuclei (blue). Regions shown at a higher magnification (far right) are indicated by yellow boxes. Endothelial cell‐specific eNOS fluorescence was measured on the luminal side of the internal elastic lamina only and expressed in mean pixel intensity per area (* P <0.05 vs the Ad.Null‐, L‐ NAME ‐, or NAM ‐treated groups). We observed that red fluorescence ( SIRT 1, eNOS ) and green fluorescence ( CD 31) signals were colocalized in the Ad. HKS group. Scale bar=50 μm. Ad.HKS indicates adenovirus containing kallistatin cDNA; Ad.Null, adenovirus containing null cDNA; DAPI, 4′,6‐diamidino‐2‐phenylindole; DHE, dihydroethidium; eNOS, endothelial nitric oxide synthase; FITC, fluoroisothiocyanate; HE, hematoxylin and eosin; L‐NAME, N ω ‐nitro‐L‐arginine methyl ester; NAM, nicotinamide; PE, phycoerythrin; SIRT1, sirtuin 1; TNF‐α, tumor necrosis factor‐α.

Article Snippet: Plasma samples were used for the analysis of TNF‐α using a Mouse TNF‐α ELISA (Proteintech, Rosemount, IL) according to the manufacturer's protocol.

Techniques: Staining, Labeling, Fluorescence, Immunostaining, Immunohistochemical staining, Microscopy, Software

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Reduced Plasma Kallistatin Is Associated With the Severity of Coronary Artery Disease, and Kallistatin Treatment Attenuates Atherosclerotic Plaque Formation in Mice

doi: 10.1161/JAHA.118.009562

Figure Lengend Snippet: Effect of kallistatin protein ( KS ) on TNF ‐α–induced HUVEC s. NADPH oxidase activity ( A ), intracellular ROS accumulation ( B ), relative SIRT 1 mRNA expression levels ( C ), relative eNOS mRNA expression levels ( D ), and SIRT 1 and eNOS protein expression ( E ). Data are presented as the mean± SEM (n=5, * P <0.05 vs the TNF ‐α, TNF ‐α+ KS + NAM , and TNF ‐α+ KS + L‐ NAME groups. # P <0.05 vs the TNF ‐α and TNF ‐α+ KS + NAM groups). eNOS indicates endothelial nitric oxide synthase; HUVEC, human umbilical vein endothelial cells; KS, kallistatin; L‐NAME, N ω ‐nitro‐L‐arginine methyl ester; NAM, nicotinamide; ROS, reactive oxygen species; SIRT1, sirtuin 1; TNF‐α, tumor necrosis factor‐α.

Article Snippet: Plasma samples were used for the analysis of TNF‐α using a Mouse TNF‐α ELISA (Proteintech, Rosemount, IL) according to the manufacturer's protocol.

Techniques: Activity Assay, Expressing

Journal:

Article Title: Analysis of Virulence and Inflammatory Potential of Shigella flexneri Purine Biosynthesis Mutants

doi: 10.1128/IAI.71.12.7002-7013.2003

Figure Lengend Snippet: Primers used in RT-PCR analysis

Article Snippet: The primary monoclonal antibodies (MAbs) employed were the following: a rat anti-mouse MHC-II MAb (Serotec, Oxford, United Kingdom), a rat anti-mouse tumor necrosis factor alpha (TNF-α) MAb (Serotec), and a murine anti- S. flexneri 5a lipopolysaccharide (LPS) MAb (6 mg/ml) ( 30 ).

Techniques: Sequencing

Journal:

Article Title: Analysis of Virulence and Inflammatory Potential of Shigella flexneri Purine Biosynthesis Mutants

doi: 10.1128/IAI.71.12.7002-7013.2003

Figure Lengend Snippet: RT-PCR and densitometry of products of RNAs extracted from lungs of three mice infected with either M90T or mutants, using primers specific for β-actin, IL-1β, IL-6, IL-12, IFN-γ, TNF-α, and iNOS at 72 h p.i. (A) Control, uninfected control lung; purHD, ZB2209 (M90T ΔpurHD); guaBA purEK, ZB504 (M90T ΔguaBA ΔpurEK); purHD purEK, ZB503 (M90T ΔpurHD ΔpurEK). (B) Control, uninfected control lung; guaBA, ZB501 (M90T ΔguaBA); guaBA purHD, ZB502 (M90T ΔguaBA ΔpurHD). Similar results were obtained in three identical experiments. Standard deviations for three experiments were within 10% of the values of the arbitrary units (a.u.).

Article Snippet: The primary monoclonal antibodies (MAbs) employed were the following: a rat anti-mouse MHC-II MAb (Serotec, Oxford, United Kingdom), a rat anti-mouse tumor necrosis factor alpha (TNF-α) MAb (Serotec), and a murine anti- S. flexneri 5a lipopolysaccharide (LPS) MAb (6 mg/ml) ( 30 ).

Techniques: Reverse Transcription Polymerase Chain Reaction, Infection

Journal:

Article Title: Analysis of Virulence and Inflammatory Potential of Shigella flexneri Purine Biosynthesis Mutants

doi: 10.1128/IAI.71.12.7002-7013.2003

Figure Lengend Snippet: Avidin-biotin immunoperoxidase labeling of TNF-α (A, B, C, and D) and of MHC-II complexes (E, F, G, and H) in tissue sections of lungs of mice infected with M90T (A and E), ZB503 (M90T ΔpurHD ΔpurEK) (B and F), ZB501 (M90T ΔguaBA) (C and G), and ZB502 (M90T ΔguaBA ΔpurHD) (D and H) at 72 h p.i. In panel A arrows point to high expression of TNF-α in neutrophil aggregates, in panels E and F arrows indicate a few MHC-II-expressing cells, in panel G arrows point to some MHC-II-expressing bronchiolar cells and arrowheads point to BALT MHC-II-positive cells, and in panel H some bronchiolar mucosal and mononuclear MHC-II-expressing cells are indicated by arrows and arrowheads, respectively. Bars, 25 μm (A, B, C, and D) and 50 μm (E, F, G, and H).

Article Snippet: The primary monoclonal antibodies (MAbs) employed were the following: a rat anti-mouse MHC-II MAb (Serotec, Oxford, United Kingdom), a rat anti-mouse tumor necrosis factor alpha (TNF-α) MAb (Serotec), and a murine anti- S. flexneri 5a lipopolysaccharide (LPS) MAb (6 mg/ml) ( 30 ).

Techniques: Avidin-Biotin Assay, Labeling, Infection, Expressing