BACKGROUND: The occurrence of portal vein system thrombosis (PVST) after splenectomy in patients with Wilson disease (WD) can lead to serious complications. The early identification of high-risk patients can help improve patient prognosis. This study aimed to establish and validate a personalized nomogram for assessing the risk of PVST after splenectomy in patients with WD and hypersplenism.
METHODS: We retrospectively collected the data from 81 patients with WD and hypersplenism who underwent splenectomy. Based on whether PVST occurred within a month after the operation, they were divided into the PVST group and the non-PVST group. The clinical data of the 2 groups were compared, and univariate analysis was used to select the statistically significant features and incorporated into the least absolute shrinkage and selection operator (LASSO) regression model for optimization. Multivariate logistic regression analysis was used to determine the independent risk factors for PVST after splenectomy, which were then applied to establish a personalized nomogram. We calculated the concordance (C)-index and drew the receiver operating characteristic (ROC) curve, the model calibration curve, and the clinical decision analysis (DCA) curve to evaluate the accuracy, calibration, and clinical applicability of the model, respectively. We used bootstrapping for internal validation of the model.
RESULTS: Univariate analysis showed that the differences in preoperative portal vein diameter and velocity of portal blood flow, postoperative mean platelet volume (MPV), mean platelet distribution width (PDW), D-dimer, prothrombin time (PT), and the increase of platelet count (PLT) were of statistical significance (P<0.05). According to the results of the LASSO and multivariate logistic regression analyses, a model including preoperative portal vein diameter, preoperative portal blood flow velocity, postoperative D-dimer, and the increase of PLT was established to predict the risk of PVST after splenectomy. The model showed good accuracy with a C-index of 0.838 (95% CI: 0.750-0.926) and had a well-fitted calibration curve. Furthermore, internal validation showed it achieved a moderate C-index of 0.805. The DCA curve indicated that the model has clinical applicability when patients are treated at thresholds of 2-100%.
CONCLUSIONS: Establishing a predictive model for the risk of PVST in patients with WD and hypersplenism after splenectomy can help clinicians identify patients at high risk of PVST who require intervention measures.

Background: Gan-Dou-Fu-Mu decoction (GDFMD) improves liver fibrosis in experimental and clinical studies including those on toxic mouse model of Wilson disease (Model). However, the mechanisms underlying the effect of GDFMD have not been characterized. Herein, we deciphered the potential therapeutic targets of GDFMD using transcriptome analysis. Methods: We constructed a tx-j Wilson disease (WD) mouse model, and assessed the effect of GDFMD on the liver of model mice by hematoxylin and eosin, Masson, and immunohistochemical staining. Subsequently, we identified differentially expressed genes (DEGs) that were upregulated in the Model (Model vs. control) and those that were downregulated upon GDFMD treatment (compared to the Model) using RNA-sequencing (RNA-Seq). Biological functions and signaling pathways in which the DEGs were involved were determined by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway analyses. A protein-protein interaction (PPI) network was constructed using the STRING database, and the modules were identified using MCODE plugin with the Cytoscape software. Several genes identified in the RNA-Seq analysis were validated by real-time quantitative PCR. Results: Total of 2124 DEGs were screened through the Model vs. control and Model vs. GDFMD comparisons, and dozens of GO and KEGG pathway terms modulated by GDFMD were identified. Dozens of pathways involved in metabolism (including metabolic processes for organic acids, carboxylic acids, monocarboxylic acids, lipids, fatty acids, cellular lipids, steroids, alcohols, eicosanoids, long-chain fatty acids), immune and inflammatory response (such as complement and coagulation cascades, cytokine-cytokine receptor interaction, inflammatory mediator regulation of TRP channels, antigen processing and presentation, T-cell receptor signaling pathway), liver fibrosis (such as ECM-receptor interactions), and cell death (PI3K-Akt signaling pathway, apoptosis, TGF-beta signaling pathway, etc.) were identified as potential targets of GDFMD in the Model. Some hub genes and four modules were identified in the PPI network. The results of real-time quantitative PCR analysis were consistent with those of RNA-Seq analysis. Conclusions: We performed gene expression profiling of GDFMD-treated WD model mice using RNA-Seq analysis and found the genes, pathways, and processes effected by the treatment. Our study provides a theoretical basis to prevent liver fibrosis resulting from WD using GDFMD.

BACKGROUND: Gandou decoction (GDD) has been widely used in the treatment of Wilson disease (WD) for decades. It is optimized from the Dahuanghuanglianxiexin decoction, Yinchenhao decoction, and Huanglianjiedu decoction. It was first reported in the Treatise on Febrile and Miscellaneous Diseases and A Handbook of Prescriptions for Emergencies published in the Eastern Han Dynasty and the Eastern Jin Dynasty respectively. Hepatic injury is one of the most severe complications of WD. The current study aimed to explore the hepatic-protection effects of GDD and its exact therapeutic target, with a particular focus on the expression of oxidative stress and the Wnt/β-catenin pathway.
METHODS: Hepatic injury was induced in a copper-loaded rat model using the intragastric administration of copper(II) sulfate pentahydrate (CuSO4·5H2O). The water extract of GDD (0.4 g/kg/d) was administered twice a day for 4 weeks. Copper content and alanine aminotransferase (ALT) level, structural observation under the microscope, and immunohistochemical analysis of liver tissue were performed after the final administration. Moreover, the expression of β-catenin, GSK3β, Dishevelled-3, c-Myc, and p-GSK3β of liver tissue were detected to explore the relationship between the hepatic protection of GDD and the Wnt/ β-catenin signal pathway of GDD. We also stimulated the rat hepatic cell line BRL-3A with CuSO4·5H2O to establish a hepatic injury cytomodel. GDD serum at a concentration of 20% was administered into the model cell for 24 h. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry were performed to detect cell viability, mitochondrial membrane potential (MMP), and the expression of reactive oxygen species (ROS). Meanwhile, the expression of the Wnt/β-catenin signal pathway-related proteins was evaluated.
RESULTS: GDD reduced copper and ALT while inhibiting oxidative stress and the degeneration and necrosis of liver tissue and hepatocytes. Treatment with GDD improved cell viability and MMP while suppressing the ROS level. Furthermore, GDD rectified the expression of Wnt/β-catenin signal pathway-related proteins in both livers of the copper-loaded and copper-stimulated BRL-3A cell lines.
CONCLUSIONS: GDD had apparent therapeutic effects on the hepatic injury of copper-loaded rats and copper-stimulated BRL-3A cells. Its mechanism is related to its regulatory effect on the Wnt/β-catenin pathway rectification and oxidative stress antagonism.