Journal: Journal of Inflammation Research

Article Title: Single-Cell Sequencing and Machine Learning Integration to Identify Candidate Biomarkers in Psoriasis: INSIG1

doi: 10.2147/JIR.S492875

Figure Lengend Snippet: Validation of INSIG1 expression in psoriasis through experiments and molecular docking results between INSIG1 and tetrandrine. ( A and B ) IHC results of INSIG1 expression in the Pso group compared with the Control group in clinical samples (n=20) and ( C and D ) in in vivo (n=6) experiments (scale bar = 50 μm and 20 μm). ( E ) mRNA expression level of INSIG1 in the Pso model group compared with the control group in vivo (n=6). ( F ) Drug sensitivity enrichment analysis of INSIG1 . ( G and H ) Visualization of molecular docking between INSIG1 and tetrandrine. ( I ) Results of interaction analysis between ligand (tetrandrine) and receptor ( INSIG1 ). Statistical significance: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, compared with the Control group.

Article Snippet: The mice were randomly assigned to four groups, that is, the Control group, the Pso model group, the tetrandrine (obtained from MedChem Express (Cat. No.:92 hY-13764; Shanghai, China) intervention group (30mg/kg, gastric lavage), and the methotrexate (MTX) as positive control group.

Techniques: Expressing, Control, In Vivo

Journal: JCI Insight

Article Title: GCN2 kinase activation mediates pulmonary vascular remodeling and pulmonary arterial hypertension

doi: 10.1172/jci.insight.177926

Figure Lengend Snippet: ( A ) Quantitative RT-PCR analysis verifying Gcn2 -specific disruption in Eif2ak4 –/– (KO) mouse lungs. WT n = 6, KO n = 7. n.d., not detected; RT, reverse transcription. ( B ) Representative micrographs of anti–phosphorylated (phospho-) GCN2 immunostaining of mouse lungs demonstrating prominent Thr898 phospho-GCN2 (GCN2-Pi) in pulmonary vascular ECs in hypoxic WT but not KO mice. Lung tissue cryosections from normoxic and hypoxic (3 weeks) WT mice and hypoxic KO mice were immunostained with anti-Thr phospho-GCN2 antibody (red). ECs were immunostained with anti-CD31 (green), and nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. ( C ) Quantification of GCN2-Pi–positive vessels. Data are expressed as percentage of positive vessels in each mouse lung. N = 5/group. ( D ) RVSP measurement showing increased RVSP in hypoxic WT mice compared with normoxic WT mice, which was reduced in hypoxic KO mice. ( E ) RV hypertrophy evident by increased RV/(LV+S) ratio seen in hypoxic WT mice was reduced in hypoxic KO mice. N = 5–8/group. Nx, normoxia; Hx, hypoxia. Data are shown as means + SD. *, P < 0.05; ****, P < 0.0001. Unpaired 2-tailed t test ( A ); 2-way ANOVA with Tukey’s multiple comparisons test ( C – E ).

Article Snippet: Fourteen days after MCT, the rats were randomized to receive either GCN2 inhibitor compound A-92 (Axon Medchem, catalog Axon 2720) (0.5 mg/kg/d, i.p.) or vehicle treatment for 14 days.

Techniques: Quantitative RT-PCR, Disruption, Reverse Transcription, Immunostaining

Journal: JCI Insight

Article Title: GCN2 kinase activation mediates pulmonary vascular remodeling and pulmonary arterial hypertension

doi: 10.1172/jci.insight.177926

Figure Lengend Snippet: ( A ) Representative micrographs of anti–phospho-GCN2 immunostaining showing GCN2 hyperphosphorylation by hypoxia challenge in primary cultures of HLMVECs. Fixed cells at indicated times after hypoxia (1% O 2 ) challenge (Hx) or normoxia (Nx) were immunostained with anti-Thr899 phospho-GCN2 (red). Nuclei were counterstained with DAPI (blue). ( B ) Quantification of cytoplasmic over nuclear GCN2-Pi at 2-hour and 4-hour hypoxia exposure. N = 5/group. ( C – G ) Western blotting demonstrating hypoxia-induced GCN2 phosphorylation and activation. HLMVECs were lysed for Western blotting with anti-Thr899 phospho-GCN2 (GCN2-Pi) antibody and anti-Ser51 phospho-EIF2α (EIF2α-Pi) antibody. Total GCN2 and EIF2α levels were assessed by anti-GCN2 antibody and anti-EIF2α antibody, respectively, while anti–β-actin was used as a loading control ( C ). The band intensities of phospho-GCN2 (GCN2-Pi) and phospho-EIF2α (EIF2α-Pi) were quantified ( D – G ). ( H – J ) Western blotting demonstrating hypoxia-induced GCN2 phosphorylation was inhibited by PDK1 inhibitor (GSK, GSK2334470) treatment in a dose-dependent manner. HLMVECs in complete growth medium were treated with GSK2334470 at 10 or 1 μM or control vehicle under 2-hour hypoxia challenge (1% O 2 ). Then cells were lysed for Western blotting with anti-Thr899 phospho-GCN2 (GCN2-Pi) antibody. Total GCN2 levels were assessed by anti-GCN2 antibody while anti-tubulin was used as a loading control ( H ). The band intensities of phospho-GCN2 were quantified ( I and J ). N = 3 repeat studies. Data are shown as means + SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Unpaired 2-tailed t test ( B ); 1-way ANOVA with Dunnett’s multiple comparisons test ( D – G ), with Holm-Šídák multiple comparisons test ( I ), or with Tukey’s multiple comparisons test ( J ).

Article Snippet: Fourteen days after MCT, the rats were randomized to receive either GCN2 inhibitor compound A-92 (Axon Medchem, catalog Axon 2720) (0.5 mg/kg/d, i.p.) or vehicle treatment for 14 days.

Techniques: Immunostaining, Western Blot, Phospho-proteomics, Activation Assay, Control

Journal: JCI Insight

Article Title: GCN2 kinase activation mediates pulmonary vascular remodeling and pulmonary arterial hypertension

doi: 10.1172/jci.insight.177926

Figure Lengend Snippet: ( A ) Representative heatmap of RNA-sequencing analysis of mouse lung tissues ( n = 4 mice combined per group). Nx, normoxia; Hx, hypoxia. ( B ) Western blotting demonstrating Edn1 protein levels were upregulated in lung tissues of hypoxic WT mice, which were significantly reduced in hypoxic KO mice. ( C ) Quantitative RT-PCR analysis verifying Edn1 mRNA upregulation in WT hypoxia mouse lungs but not in KO hypoxia mouse lungs. N = 5–8/group. ( D ) Quantitative RT-PCR analysis demonstrating GCN2 siRNA–mediated (siGCN2-mediated) knockdown of GCN2 in HLMVECs. N = 5/group. CTL, control siRNA. ( E ) Quantitative RT-PCR analysis demonstrating hypoxia-induced EDN1 mRNA expression was mediated by GCN2 in HLMVECs. N = 6/group. ( F ) ELISA of EDN1 secreted in culture medium demonstrating hypoxia-induced EDN1 protein expression was reduced by GCN2 silencing in HLMVECs. N = 6/group. ( G ) Western blotting confirmation of reduced HIF-2α expression in GCN2-deficient HLMVECs compared with control cells under hypoxia exposure. ( H ) Quantitative RT-PCR analysis demonstrating inhibited HIF2A mRNA expression in GCN2-deficient HLMVECs. Control n = 6, siGCN2 n = 8. ( I ) Quantitative RT-PCR analysis demonstrating GCN2 mediated hypoxia-induced EDN1 expression through HIF-2α rather than HIF-1α. HLMVECs were transfected with either siGCN2 or control siRNA (CtlsiRNA), and HIF1A , HIF2A , or vector plasmid, and then challenged with hypoxia or maintained in normoxia. At 48 hours after hypoxia challenge or normoxia, the cells were lysed for RNA isolation for quantitative RT-PCR analysis. N = 6–10/group. Data are shown as means + SD. *, P < 0.05; ****, P < 0.0001. Unpaired 2-tailed t test ( D and H ); 2-way ANOVA with Tukey’s multiple comparisons test ( C , E , and F ); 1-way ANOVA with Tukey’s multiple comparisons test ( I ).

Article Snippet: Fourteen days after MCT, the rats were randomized to receive either GCN2 inhibitor compound A-92 (Axon Medchem, catalog Axon 2720) (0.5 mg/kg/d, i.p.) or vehicle treatment for 14 days.

Techniques: RNA Sequencing, Western Blot, Quantitative RT-PCR, Knockdown, Control, Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Isolation

Journal: JCI Insight

Article Title: GCN2 kinase activation mediates pulmonary vascular remodeling and pulmonary arterial hypertension

doi: 10.1172/jci.insight.177926

Figure Lengend Snippet: ( A ) Diagram showing the experimental procedure to restore Edn1 expression selectively in ECs of WT and Gcn2 -deficient mice. Mixture of nanoparticles/plasmid DNA expressing Edn1 or GFP (control) under the control of CDH5 promoter was administered retro-orbitally to 11-week-old KO mice and WT mice. After overnight (16 hours), the mice were subjected to hypoxia. Nanoparticles/plasmid DNA mixtures were administered weekly for 3 doses total, and each mouse received 30 μg plasmid DNA each time. ( B ) Western blotting demonstrating restored Edn1 expression in ECs of KO mice with Edn1 plasmid administration compared with WT and KO mice with GFP plasmid. Lung ECs were isolated for Western blotting. ( C ) RVSP measurement showing reduced PH in KO mice with GFP plasmid was partially reversed in KO mice with Edn1 plasmid. ( D ) RV/(LV+S) ratio also showing reduced RV hypertrophy in KO mice with GFP plasmid was partially rescued with restored Edn1 expression. N = 5–8/group. ( E ) Representative micrographs of anti–α-SMA staining of lung sections showing reduced number of muscularized distal pulmonary vessels in hypoxic KO+ GFP lungs was partially reversed in hypoxic KO+ Edn1 lungs. Nuclei were counterstained with DAPI (blue). Arrows point to muscularized vessels. ( F ) Quantification of the number of muscularized distal pulmonary vessels. N = 5/group. ( G ) Representative micrographs of Russell-Movat pentachrome staining of mouse lung sections. Br, bronchiole; V, vessel. ( H ) Quantification of pulmonary vessel media wall thickness. N = 5/group. Scale bars, 50 μm. Data are shown as means + SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. One-way ANOVA with Tukey’s multiple comparisons test ( C , D , F , and H ).

Article Snippet: Fourteen days after MCT, the rats were randomized to receive either GCN2 inhibitor compound A-92 (Axon Medchem, catalog Axon 2720) (0.5 mg/kg/d, i.p.) or vehicle treatment for 14 days.

Techniques: Expressing, Plasmid Preparation, Control, Western Blot, Isolation, Staining

Journal: JCI Insight

Article Title: GCN2 kinase activation mediates pulmonary vascular remodeling and pulmonary arterial hypertension

doi: 10.1172/jci.insight.177926

Figure Lengend Snippet: ( A ) Representative micrographs of anti–phospho-GCN2 immunostaining of rat lung sections demonstrating prominent Thr898 phosphorylation-GCN2 (GCN2-Pi) in pulmonary vascular ECs in vehicle-treated MCT rats, which was inhibited in A-92–treated MCT rat lungs. Lung tissue cryosections from basal control rats, vehicle-treated MCT rats (4 weeks), or compound A-92–treated MCT rats were immunostained with anti-Thr898 phospho-GCN2 antibody (red). ECs were immunostained with anti-CD31 (green), and nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. ( B ) Graphical presentation of the experimental procedure. A-92 (0.5 mg/kg, i.p. daily) was administered to rats at 2 weeks after MCT. ( C ) RVSP measurement showing a marked increase of RVSP in MCT rats treated with vehicle compared with basal rats, which was reduced in A-92–treated MCT rats. ( D ) RV hypertrophy evident by increased RV/(LV+S) ratio seen in vehicle MCT rats was reduced in A-92 MCT rats. Basal n = 5, MCT veh n = 10, MCT A92 n = 10. ( E ) Representative micrographs of Russell-Movat pentachrome staining of rat lung sections showing attenuated vessel wall thickening. Br, bronchiole; V, vessel. ( F ) Quantification of average pulmonary vessel wall thickness. N = 5/group. MWT, media wall thickness. ( G ) Representative micrographs of anti–α-SMA (green) staining of basal rat, MCT vehicle rat, and A-92–treated MCT rat lung sections showing reduced number of muscularized distal pulmonary vessels in A-92–treated MCT rat lungs. Nuclei were counterstained with DAPI (blue). Arrows point to muscularized distal pulmonary vessels. ( H ) Quantification of muscularized distal pulmonary vessels. The total number of α-SMA–positive distal pulmonary vessels (d ≤ 50 μm) of 20× original magnification fields of each section was used for each rat. N = 5–7/group. Arrows point to muscularized vessels. Data are shown as means + SD. Scale bars, 50 μm. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. One-way ANOVA with Tukey’s multiple comparisons test ( C , D , F , and H ).

Article Snippet: Fourteen days after MCT, the rats were randomized to receive either GCN2 inhibitor compound A-92 (Axon Medchem, catalog Axon 2720) (0.5 mg/kg/d, i.p.) or vehicle treatment for 14 days.

Techniques: Immunostaining, Phospho-proteomics, Control, Staining

Journal: JCI Insight

Article Title: GCN2 kinase activation mediates pulmonary vascular remodeling and pulmonary arterial hypertension

doi: 10.1172/jci.insight.177926

Figure Lengend Snippet: ( A and B ) Western blotting of anti-GCN2 and quantification demonstrating no difference in GCN2 total protein expression in whole lung tissue lysates of patients with IPAH compared with normal donors. Control n = 5, patients with IPAH n = 7. ( C and D ) Western blotting of anti-GCN2 and quantification demonstrating no difference in GCN2 total protein expression in pulmonary arterial ECs of patients with IPAH compared with healthy donors. Control n = 4, patients with IPAH n = 5. ( E ) Western blotting demonstrating extensive GCN2 phosphorylation in IPAH patient lung tissues but minimal in control donor lung tissues. ( F and G ) Prominent GCN2 phosphorylation in ECs of pulmonary vascular lesions of patients with IPAH but not in normal donor lungs. Formalin-fixed lung sections from 5 patients with IPAH and 5 non-PAH donors were immunostained with anti-Thr899 phospho-GCN2 (GCN2-Pi) (red) and anti-vWF (green). Representative micrographs of immunofluorescence staining were shown ( F ). The average number of GCN2-Pi + ECs in each vessel was quantified ( n = 5 samples/group, 15 vessels/sample) ( G ). Data are shown as means + SD. Scale bar, 50 μm. **, P < 0.01. Mann-Whitney U test ( B ). Unpaired 2-tailed t test ( D and G ).

Article Snippet: Fourteen days after MCT, the rats were randomized to receive either GCN2 inhibitor compound A-92 (Axon Medchem, catalog Axon 2720) (0.5 mg/kg/d, i.p.) or vehicle treatment for 14 days.

Techniques: Western Blot, Expressing, Control, Phospho-proteomics, Immunofluorescence, Staining, MANN-WHITNEY

Journal: Ontario Health Technology Assessment Series

Article Title: Cervical Artificial Disc Replacement Versus Fusion for Cervical Degenerative Disc Disease: A Health Technology Assessment

doi:

Figure Lengend Snippet: Randomized Controlled Trials of Cervical Artificial Disc Replacement Versus Fusion for Cervical Degenerative Disc Disease

Article Snippet: Health care resource use and QALYs were prospectively obtained from the randomized, multicentre study and post-approval study of ProDisc-C total disc replacement for IDE (Murrey et al, 22 2009; Janssen et al, 92 2015).

Techniques:

Journal: Ontario Health Technology Assessment Series

Article Title: Cervical Artificial Disc Replacement Versus Fusion for Cervical Degenerative Disc Disease: A Health Technology Assessment

doi:

Figure Lengend Snippet: Cervical Implant Devices Used in Randomized Controlled Trials of C-ADR Versus Fusion for Degenerative Disc Disease

Article Snippet: Health care resource use and QALYs were prospectively obtained from the randomized, multicentre study and post-approval study of ProDisc-C total disc replacement for IDE (Murrey et al, 22 2009; Janssen et al, 92 2015).

Techniques: Functional Assay

Journal: Ontario Health Technology Assessment Series

Article Title: Cervical Artificial Disc Replacement Versus Fusion for Cervical Degenerative Disc Disease: A Health Technology Assessment

doi:

Figure Lengend Snippet: Cervical Motion at Treated and Adjacent Sites After C-ADR or Fusion

Article Snippet: Health care resource use and QALYs were prospectively obtained from the randomized, multicentre study and post-approval study of ProDisc-C total disc replacement for IDE (Murrey et al, 22 2009; Janssen et al, 92 2015).

Techniques: Software

Journal: Ontario Health Technology Assessment Series

Article Title: Cervical Artificial Disc Replacement Versus Fusion for Cervical Degenerative Disc Disease: A Health Technology Assessment

doi:

Figure Lengend Snippet: Secondary Surgeries in Longer-Term Follow-Up of C-ADR– Versus Fusion–Treated One-Level Cervical Degenerative Disc Disease

Article Snippet: Health care resource use and QALYs were prospectively obtained from the randomized, multicentre study and post-approval study of ProDisc-C total disc replacement for IDE (Murrey et al, 22 2009; Janssen et al, 92 2015).

Techniques: Migration

Journal: European Radiology

Article Title: Normalizing computed tomography data reconstructed with different filter kernels: effect on emphysema quantification

doi: 10.1007/s00330-015-3824-y

Figure Lengend Snippet: Example sections of a scan reconstructed with two kernels in the Siemens scanner with an emphysema overlay highlighting voxels below -950HU. The upper row shows the original sections reconstructed with ( A ) b31f and ( B ) b45f kernels. The lower row shows the same sections after normalization with ( C ) b31f and ( D ) b45f kernels. The ES obtained for the ( A ) b31f original scan was 9.5 %, for the ( B ) b45f original, ES was 21.2 %, whereas for the ( C ) b31f normalized, ES was 10.1 % and for the ( D ) b45f normalized, ES was 10.9 %

Article Snippet: The second mixed cohort (n = 369) was constructed to contain scans with both standard and sharp kernels from both manufacturers: from the Siemens group, the b31f reconstruction was chosen for 92 randomly selected subjects, and the b45f reconstruction for the remaining 91 subjects.

Techniques:

Journal: European Radiology

Article Title: Normalizing computed tomography data reconstructed with different filter kernels: effect on emphysema quantification

doi: 10.1007/s00330-015-3824-y

Figure Lengend Snippet: Bland-Altman plots comparing ES values for (A) original b31f and original b45f, and (B) normalized b31f and normalized b45f in the Siemens group. The mean differences are shown with a solid line ; the limits of agreement are shown with dashed lines

Article Snippet: The second mixed cohort (n = 369) was constructed to contain scans with both standard and sharp kernels from both manufacturers: from the Siemens group, the b31f reconstruction was chosen for 92 randomly selected subjects, and the b45f reconstruction for the remaining 91 subjects.

Techniques:

Journal: European Radiology

Article Title: Normalizing computed tomography data reconstructed with different filter kernels: effect on emphysema quantification

doi: 10.1007/s00330-015-3824-y

Figure Lengend Snippet: Correlation of ES measured for the three groups (Siemens, GE and Mixed), for each cohort, before and after normalization

Article Snippet: The second mixed cohort (n = 369) was constructed to contain scans with both standard and sharp kernels from both manufacturers: from the Siemens group, the b31f reconstruction was chosen for 92 randomly selected subjects, and the b45f reconstruction for the remaining 91 subjects.

Techniques: