Fig. 1

Identification of THBS1 as a potential key biomarker in HBV-ACLF pathogenesis. A Schematic representation of the study design and analysis strategy flow chart. To identify potential biomarkers, biobanked peripheral blood mononuclear cells (PBMCs) collected from patients in the Chinese Group on the Study of Severe Hepatitis B (COSSH) multicenter cohort underwent transcriptomic sequencing (ACLF = 20; LC = 10; CHB = 10; NC = 15), and the results were validated in humans, an ACLF preclinical rat model and THBS1^KO ALF mice. B Calculated frequencies of the top 10 genes (ranked by adjusted P value) that were identified in the multigroup comparison. C The top 10 genes are shown in the volcano plot of the ACLF vs. NC groups. The vertical dashed lines indicate the threshold for the fourfold difference in abundance. The horizontal dashed line indicates the adjusted P value = 0.05 threshold. Red represents significantly upregulated genes (THBS1 in green), while blue represents significantly downregulated genes. The node size represents the frequency of key genes. D-F THBS1 expression profile in the sequencing group (D), and RT–qPCR validation of THBS1 expression (E) in human PBMCs (ACLF = 110; LC = 60; CHB = 60; NC = 45). *** P value < 0.001, GAPDH was set as a reference control gene. Serum levels of THBS1 measured by ELISA (F) and analyzed using the Mann–Whitney U test (ACLF = 198; LC = 50; CHB = 50; NC = 50). *** P value < 0.001 vs. NC group. G Representative immunohistochemical staining of THBS1 in liver tissues (black arrow) from patients in the HBV-ACLF, LC, CHB and NC groups. H Quantitative multiplex tyramide signal amplification (TSA) immunofluorescent images showing THBS1 expression intensities in liver tissues from patients in the HBV-ACLF, LC and NC groups, stained with THBS1 (red), a hepatocyte marker (ALB, purple) and a macrophage marker (CD86, green) and a nuclear marker (DAPI, blue)