Fig 1: Peripheral blood mononuclear cell transcriptomic profiling identifying the VSIG4 expression level as strongly associated with poor ACLF outcomes.(A) Volcano plot depicting DEGs between ACLF non-survivors and survivors in the sequencing derivation cohort. Significant DEGs (|log2 fold change| >2; adjusted p value <0.05) are shown in red or blue. (B) Boxplots showing VSIG4 mRNA expression levels in the ACLF non-survivor, survivor, CLD and healthy groups. Level of significance: adj p < 0.05 (Wald test and corrected by Benjamini?Hochberg procedure). (C) Gene set-enrichment analysis results based on the rank-ordered gene list according to the Pearson’s correlation coefficient with VSIG4 mRNA level. The top 10 significantly enriched gene ontology biological processes are shown in each plot. ACLF, acute-on-chronic liver failure; BTMs, blood transcriptional modules; DEGs, differentially expressed genes; GO, gene ontology; NES, normalized enrichment score; NK, natural killer; WBMs, whole blood modules.
Fig 2: Validation of VSIG4 expression levels in patients with non-HBV-ACLF.(A) Boxplots showing plasma VSIG4 expression levels stratified by non-HBV-ACLF outcomes at 28 days, and the presence of coagulation and lung failure. Level of significance: p < 0.05 (Mann?Whitney test). (B) ROC curves of plasma VSIG4 expression levels for predicting the 28-day (left) and 90-day (right) mortality of patients with non-HBV-ACLF compared to three clinical scores. Level of significance: p < 0.05 (DeLong test). (C) Kaplan?Meier curves of survival probability stratified by plasma VSIG4 level at 28 days (left) and 90 days (right). Level of significance: p < 0.05 (log-rank test). The number of patients at risk is shown for each observation period. ACLF, acute-on-chronic liver failure; COSSH-ACLF, Chinese Group on the Study of Severe Hepatitis B-ACLF; CLIF-C ACLF, Chronic Liver Failure-Consortium ACLF; MELD-Na, model for end-stage liver disease-sodium.
Fig 3: Prognostic accuracy and risk stratification of plasma VSIG4 levels.(A) ROC curves of plasma VSIG4 expression levels for predicting the 28-day (left) and 90-day (right) mortality of patients with ACLF compared to three clinical scores. Level of significance: p < 0.05 (DeLong test). (B) The prognostic score formulas according to the multivariate Cox proportional hazards model. (C) Kaplan?Meier curves of survival probability stratified by plasma VSIG4 level at 28 days (left) and 90 days (right). Level of significance: p < 0.05 (log-rank test). The number of patients at risk is shown for each observation period. ACLF, acute-on-chronic liver failure; COSSH-ACLF, Chinese Group on the Study of Severe Hepatitis B-ACLF; CLIF-C ACLF, Chronic Liver Failure-Consortium ACLF; MELD-Na, model for end-stage liver disease-sodium; PH, proportional hazards.
Fig 4: Validation of VSIG4 expression levels in ACLF.(A) Peripheral blood mononuclear cell VSIG4 expression levels were tested by qRT?PCR in the same patients analyzed for the sequencing cohort. Level of significance: p < 0.05 (Kruskal?Wallis test. ANOVA). (B) Plasma VSIG4 expression levels in patients at different disease stages tested by ELISA. Level of significance: p < 0.05 (Kruskal?Wallis test. ANOVA). (C) ROC curves of plasma VSIG4 expression levels for distinguishing patients with ACLF from those with CLD and healthy controls. (D) Boxplots showing plasma VSIG4 expression levels stratified by ACLF grade. Level of significance: p < 0.05 (Kruskal?Wallis test. ANOVA) (E) Boxplots showing plasma VSIG4 expression levels stratified by the presence of organ failure. Level of significance: p < 0.05 (Mann?Whitney test). ACLF, acute-on-chronic liver failure; CLD, chronic liver disease; qRT-PCR, quantitative reverse-transcription PCR.
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