Journal: Health Science Reports

Article Title: CD40LG as a Biomarker in Rheumatoid Arthritis: Links to Bone Destruction and Interstitial Lung Disease ‐ A Bioinformatic Analysis With Clinical Validation

doi: 10.1002/hsr2.71318

Figure Lengend Snippet: Comparison of the expression of genes in RA and RA synovial tissues. (A) There was no difference in CXCL 8 gene expression between RA and OA patients ( p = 0.097). (B) The CD40LG gene expression was higher in RA patients than in OA patients ( p = 0.002). (C) The ITGAX gene expression was higher in RA patients than in OA patients( p = 0.025). (D) There was no difference in IL6 gene expression between RA and OA patients( p = 0.687). (E)The PTPRC gene expression was higher in RA patients than in OA patients( p = 0.003). * p < 0.05; ** p < 0.01.

Article Snippet: The recombinant CD40LG (Novoprotein), ITGAX (Proteintech), and PTPRC (Novoprotein) proteins (0.1 μg/mL) were used to coat 96‐well plates overnight at 4°C, followed by blocking 5% BSA at 37°C for 1 h. After washing three times with PBST, 100 μl of 1:100 dilution serum was added to each well and incubated plates at 37°C for 1 h. After another triple PBST rinse, peroxidase‐conjugated affinipure goat anti‐human IgG (1:10,000; Jackson ImmunoResearch) was added to the wells and incubated at 37°C for 1 h. After triple PBST washes, add the color development solution, once the color appears, immediately apply the stop solution to terminate the reaction.

Techniques: Comparison, Expressing, Gene Expression

Journal: Health Science Reports

Article Title: CD40LG as a Biomarker in Rheumatoid Arthritis: Links to Bone Destruction and Interstitial Lung Disease ‐ A Bioinformatic Analysis With Clinical Validation

doi: 10.1002/hsr2.71318

Figure Lengend Snippet: Receiver operating characteristic (ROC) curve of the 5 specifically expressed hub genes. (A) ROC curves for CXCL8. (B) ROC curves for CD40LG. (C) ROC curves for PTPRC. (D) ROC curves for IL6. (E) ROC curves for ITGAX. (F) 5 genes ROC summary.

Article Snippet: The recombinant CD40LG (Novoprotein), ITGAX (Proteintech), and PTPRC (Novoprotein) proteins (0.1 μg/mL) were used to coat 96‐well plates overnight at 4°C, followed by blocking 5% BSA at 37°C for 1 h. After washing three times with PBST, 100 μl of 1:100 dilution serum was added to each well and incubated plates at 37°C for 1 h. After another triple PBST rinse, peroxidase‐conjugated affinipure goat anti‐human IgG (1:10,000; Jackson ImmunoResearch) was added to the wells and incubated at 37°C for 1 h. After triple PBST washes, add the color development solution, once the color appears, immediately apply the stop solution to terminate the reaction.

Techniques:

Journal: Health Science Reports

Article Title: CD40LG as a Biomarker in Rheumatoid Arthritis: Links to Bone Destruction and Interstitial Lung Disease ‐ A Bioinformatic Analysis With Clinical Validation

doi: 10.1002/hsr2.71318

Figure Lengend Snippet: Comparison of the relevant antibody levels in the serum of different patients. (A) Comparison of OD values of serum CD40LG, ITGAX and PTPRC in RA, OA, and the HC group. (B) ROC curve of CD40LG in RA patients. (C) Higher serum CD40LG antibody levels in RA patients with combined bone destruction and ILD. ns: no significant difference. * p < 0.05; **** p < 0.0001. ROC, receiver operating characteristic.

Article Snippet: The recombinant CD40LG (Novoprotein), ITGAX (Proteintech), and PTPRC (Novoprotein) proteins (0.1 μg/mL) were used to coat 96‐well plates overnight at 4°C, followed by blocking 5% BSA at 37°C for 1 h. After washing three times with PBST, 100 μl of 1:100 dilution serum was added to each well and incubated plates at 37°C for 1 h. After another triple PBST rinse, peroxidase‐conjugated affinipure goat anti‐human IgG (1:10,000; Jackson ImmunoResearch) was added to the wells and incubated at 37°C for 1 h. After triple PBST washes, add the color development solution, once the color appears, immediately apply the stop solution to terminate the reaction.

Techniques: Comparison

Journal: Health Science Reports

Article Title: CD40LG as a Biomarker in Rheumatoid Arthritis: Links to Bone Destruction and Interstitial Lung Disease ‐ A Bioinformatic Analysis With Clinical Validation

doi: 10.1002/hsr2.71318

Figure Lengend Snippet: Molecular docking 3D diagram of key targets and small molecule drugs. (A) CD40LG‐emetine complex, with binding energy of −7.1 kcal/mol. (B) CD40LG‐oligomycin complex, with binding energy of −7.2 kcal/mol. (C) CD40LG‐cromoglicic acid complex, with binding energy of −7.9 kcal/mol. (D) ITGAX‐sulfinpyrazone complex, with binding energy of −7.2 kcal/mol. (E) PTPRC‐oligomycin complex, with binding energy of −8.5 kcal/mol. (F) PTPRC‐buspirone complex, with binding energy of −7.0 kcal/mol. (G) PTPRC‐chlorogenic acid complex, with binding energy of −7.2 kcal/mol.

Article Snippet: The recombinant CD40LG (Novoprotein), ITGAX (Proteintech), and PTPRC (Novoprotein) proteins (0.1 μg/mL) were used to coat 96‐well plates overnight at 4°C, followed by blocking 5% BSA at 37°C for 1 h. After washing three times with PBST, 100 μl of 1:100 dilution serum was added to each well and incubated plates at 37°C for 1 h. After another triple PBST rinse, peroxidase‐conjugated affinipure goat anti‐human IgG (1:10,000; Jackson ImmunoResearch) was added to the wells and incubated at 37°C for 1 h. After triple PBST washes, add the color development solution, once the color appears, immediately apply the stop solution to terminate the reaction.

Techniques: Binding Assay

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 1: CD40LG is down-regulated in multiple cancer types, and this down-regulation has significant prognostic implications. (A) Violin plots showed the differential expression of CD40LG in 34 cancer types compared with normal tissues. (B) Body map showing CD40LG expression levels in various normal human tissues. (C) Cox regression analysis of OS demonstrated the prognostic role of CD40LG in a variety of cancers. (D) Cox regression analysis of PFS demonstrated the prognostic role of CD40LG in a variety of cancers. (E) The veen diagram shows those cancers in which CD40LG has the same prognostic predictive efficacy in OS and PFS.

Article Snippet: Subsequently, they were incubated with the primary antibody against GAPDH (1:1,000, 60004-1-lg, Proteintech) and CD40LG (1:1,000, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Quantitative Proteomics, Expressing

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 2: Correlation Between CD40LG Expression and Immunotherapeutic Efficacy of Various Cancers. (A) Kaplan-Meier plotter analysis showing that patients with high CD40LG expression have better OS after immunotherapy. (B) Kaplan-Meier plotter analysis showing that patients with high CD40LG expression have better PFS after immunotherapy. (C) Comparison of CD40LG expression with traditional biomarkers including PD-L1 expression, TMB, MSI status and CD8+T cell density in predicting immunotherapy outcomes across 25 datasets.

Article Snippet: Subsequently, they were incubated with the primary antibody against GAPDH (1:1,000, 60004-1-lg, Proteintech) and CD40LG (1:1,000, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Comparison

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 3: The important role of CD40LG expression in LUAD. (A-C) Reduced expression of CD40LG in LUAD was analysed in public databases, including TCGA, GSE118370 and GSE43458. (D-F) The association of CD40LG expression with overall survival of LUAD patients was examined in public databases, including GSE30219, GSE31210 and GSE72094. (G) CD40LG expression in LUADs analysed in immunohistochemistry of HPA was lower than in corresponding paraneoplastic tissues. (H) PCR experiments were performed on four patients with LUAD to verify the RNA expression of CD40LG in cancer and corresponding paracancerous tissues. (I) Western blot experiments were performed on four patients with LUAD to verify the protein expression of CD40LG in cancer and corresponding paracancerous tissues.

Article Snippet: Subsequently, they were incubated with the primary antibody against GAPDH (1:1,000, 60004-1-lg, Proteintech) and CD40LG (1:1,000, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Immunohistochemistry, RNA Expression, Western Blot

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 4: CD40LG Expression Correlates with Immune Microenvironment in LUAD. (A) Correlation between CD40LG expression and Immunophenoscore (IPS), including components such as antigen presentation, effector cells, suppressor cells, and immune checkpoints in LUAD. (B-F) Positive correlation of CD40LG expression with infiltration of various immune cells in LUAD tissues, including CD8+ T cells, dendritic cells, macrophages, B cells, and CD4+ T cells, using multiple predictive methods.

Article Snippet: Subsequently, they were incubated with the primary antibody against GAPDH (1:1,000, 60004-1-lg, Proteintech) and CD40LG (1:1,000, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Immunopeptidomics

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 5: CD40LG Expression in LUAD and its Association with Clinical Stages. (A) Representative immunohistochemical staining images of CD40LG in primary LUAD tissues and corresponding paracancerous tissues. (B) Boxplot comparing CD40LG expression levels in tumor tissues versus paracancerous tissues. (C-D) The boxplot analysis illustrates the expression of CD40LG in relation to TNM and T stages.

Article Snippet: Subsequently, they were incubated with the primary antibody against GAPDH (1:1,000, 60004-1-lg, Proteintech) and CD40LG (1:1,000, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 6: CD40LG Expression Correlated with Immune Infiltrate Patterns in LUAD. (A) Representative mIF staining image showing CD40LG expression and immune cell distribution in LUAD tissue microarrays. (B-E) Correlation analysis between CD40LG_IHC_Score and percentages of various immune-related cells in total, tumor, and stromal regions.

Article Snippet: Subsequently, they were incubated with the primary antibody against GAPDH (1:1,000, 60004-1-lg, Proteintech) and CD40LG (1:1,000, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Staining

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 7: CD40LG Expression and Its Effects on LUAD Cell Migration, Invasion, and Proliferation. (A) Western blot analysis of CD40LG expression in five LUAD cell lines. (B) Western blot verification of siRNA-mediated knockdown of CD40LG in H1975 cells and overexpression in A549 cells. (C) EDU assay results showing increased proliferation upon CD40LG knockdown in H1975 cells and decreased proliferation with CD40LG overexpression in A549 cells. (D) Scratch wound healing assay indicating enhanced migration with CD40LG knockdown in H1975 cells and reduced migration with CD40LG overexpression in A549 cells. (E) Transwell assay demonstrating increased migration and invasion capabilities with CD40LG knockdown in H1975 cells and decreased capabilities with CD40LG overexpression in A549 cells.

Article Snippet: Subsequently, they were incubated with the primary antibody against GAPDH (1:1,000, 60004-1-lg, Proteintech) and CD40LG (1:1,000, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Migration, Western Blot, Knockdown, Over Expression, EdU Assay, Wound Healing Assay, Transwell Assay

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 1: CD40LG is down-regulated in multiple cancer types, and this down-regulation has significant prognostic implications. (A) Violin plots showed the differential expression of CD40LG in 34 cancer types compared with normal tissues. (B) Body map showing CD40LG expression levels in various normal human tissues. (C) Cox regression analysis of OS demonstrated the prognostic role of CD40LG in a variety of cancers. (D) Cox regression analysis of PFS demonstrated the prognostic role of CD40LG in a variety of cancers. (E) The veen diagram shows those cancers in which CD40LG has the same prognostic predictive efficacy in OS and PFS.

Article Snippet: Afterwards, they were incubated with the primary antibody against CD40LG (1:150, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Quantitative Proteomics, Expressing

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 2: Correlation Between CD40LG Expression and Immunotherapeutic Efficacy of Various Cancers. (A) Kaplan-Meier plotter analysis showing that patients with high CD40LG expression have better OS after immunotherapy. (B) Kaplan-Meier plotter analysis showing that patients with high CD40LG expression have better PFS after immunotherapy. (C) Comparison of CD40LG expression with traditional biomarkers including PD-L1 expression, TMB, MSI status and CD8+T cell density in predicting immunotherapy outcomes across 25 datasets.

Article Snippet: Afterwards, they were incubated with the primary antibody against CD40LG (1:150, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Comparison

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 3: The important role of CD40LG expression in LUAD. (A-C) Reduced expression of CD40LG in LUAD was analysed in public databases, including TCGA, GSE118370 and GSE43458. (D-F) The association of CD40LG expression with overall survival of LUAD patients was examined in public databases, including GSE30219, GSE31210 and GSE72094. (G) CD40LG expression in LUADs analysed in immunohistochemistry of HPA was lower than in corresponding paraneoplastic tissues. (H) PCR experiments were performed on four patients with LUAD to verify the RNA expression of CD40LG in cancer and corresponding paracancerous tissues. (I) Western blot experiments were performed on four patients with LUAD to verify the protein expression of CD40LG in cancer and corresponding paracancerous tissues.

Article Snippet: Afterwards, they were incubated with the primary antibody against CD40LG (1:150, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Immunohistochemistry, RNA Expression, Western Blot

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 4: CD40LG Expression Correlates with Immune Microenvironment in LUAD. (A) Correlation between CD40LG expression and Immunophenoscore (IPS), including components such as antigen presentation, effector cells, suppressor cells, and immune checkpoints in LUAD. (B-F) Positive correlation of CD40LG expression with infiltration of various immune cells in LUAD tissues, including CD8+ T cells, dendritic cells, macrophages, B cells, and CD4+ T cells, using multiple predictive methods.

Article Snippet: Afterwards, they were incubated with the primary antibody against CD40LG (1:150, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Immunopeptidomics

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 5: CD40LG Expression in LUAD and its Association with Clinical Stages. (A) Representative immunohistochemical staining images of CD40LG in primary LUAD tissues and corresponding paracancerous tissues. (B) Boxplot comparing CD40LG expression levels in tumor tissues versus paracancerous tissues. (C-D) The boxplot analysis illustrates the expression of CD40LG in relation to TNM and T stages.

Article Snippet: Afterwards, they were incubated with the primary antibody against CD40LG (1:150, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 6: CD40LG Expression Correlated with Immune Infiltrate Patterns in LUAD. (A) Representative mIF staining image showing CD40LG expression and immune cell distribution in LUAD tissue microarrays. (B-E) Correlation analysis between CD40LG_IHC_Score and percentages of various immune-related cells in total, tumor, and stromal regions.

Article Snippet: Afterwards, they were incubated with the primary antibody against CD40LG (1:150, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Staining

Journal: Journal of Cancer

Article Title: CD40LG Downregulation in Lung Adenocarcinoma: A Prognostic Biomarker Linked to Immune Cell Infiltration and Survival Outcomes

doi: 10.7150/jca.115525

Figure Lengend Snippet: Figure 7: CD40LG Expression and Its Effects on LUAD Cell Migration, Invasion, and Proliferation. (A) Western blot analysis of CD40LG expression in five LUAD cell lines. (B) Western blot verification of siRNA-mediated knockdown of CD40LG in H1975 cells and overexpression in A549 cells. (C) EDU assay results showing increased proliferation upon CD40LG knockdown in H1975 cells and decreased proliferation with CD40LG overexpression in A549 cells. (D) Scratch wound healing assay indicating enhanced migration with CD40LG knockdown in H1975 cells and reduced migration with CD40LG overexpression in A549 cells. (E) Transwell assay demonstrating increased migration and invasion capabilities with CD40LG knockdown in H1975 cells and decreased capabilities with CD40LG overexpression in A549 cells.

Article Snippet: Afterwards, they were incubated with the primary antibody against CD40LG (1:150, 16668-1-AP, Proteintech) at 4°C overnight.

Techniques: Expressing, Migration, Western Blot, Knockdown, Over Expression, EdU Assay, Wound Healing Assay, Transwell Assay