Serum HMGB-1 released by ferroptosis and necroptosis as a novel potential biomarker for systemic lupus erythematosus.

High mobility group box proterin-1 (HMGB-1) is a multifunctional protein that can be released by various programmed cell deaths (PCDs), such as necroptosis and ferroptosis. PCDs play a critical role in the pathogenesis of systemic lupus erythematosus (SLE). However, the role of HMGB-1 in the process of SLE remains unclear. This study aims to demonstrate the potential diagnosing role of serum HMGB-1 in SLE that released by necroptosis and ferroptosis. We found that the serum levels of HMGB-1, receptor-interacting protein kinase 3 (RIPK3) /mixed lineage kinase domain-like protein (MLKL) related with necroptosis, and metabolites associated with ferroptosis were significantly upregulated in SLE patients compared to HC individuals. These serum levels were positively correlated with SLE disease activity. Additionally, the serum level of HMGB-1 showed a strong positive correlated with the levels of RIPK3/MLKL and ferroptosis metabolites. Moreover, the serum level of HMGB-1 was correlated with renal involvement and high-antinuclear antibodies (ANA) titer. After SLE serum and interferon γ (IFN-γ) treatment in vitro, the level of necroptosis and ferroptosis markers were activated and HMGB1 was released both in HEK293 and HK2 cells. Clinically, HMGB-1 was considered as a significant independent risk factor in SLE serum by binary logistic assay. Notably, HMGB-1 exhibited outstanding diagnostic ability for SLE by the area under the curve (AUC) in receiver operating characteristic (ROC) curve analysis. Taken together, our study indicates that the serum level of HMGB-1 is a promising biomarker for the diagnosis and monitoring of SLE.

Construction of diagnostic prediction model for Wilson's disease.

Background: Wilson's disease, also known as hepatolenticular degeneration, is a rare human autosomal recessive inherited disorder of copper metabolism. The clinical manifestations are diverse, and the diagnosis and treatment are often delayed. The purpose of this study is to establish a new predictive diagnostic model of Wilson's disease and evaluate its predictive efficacy by multivariate regression analysis of small trauma, good accuracy, low cost, and quantifiable serological indicators, in order to identify Wilson's disease early, improve the diagnosis rate, and clarify the treatment plan. Methods: A retrospective analysis was performed on 127 patients with Wilson's disease admitted to the First People's Hospital of Yunnan Province from January 2003 to May 2022 as the experimental group and 73 patients with normal serological indicators who were not diagnosed with Wilson's disease. SPSS version 26.0 software was used for single factor screening and a multivariate binary logistic regression analysis to screen out independent factors. R version 4.1.0 software was used to establish an intuitive nomogram prediction model for the independent influencing factors included. The accuracy of the nomogram prediction model was evaluated and quantified by calculating the concordance index (C-index) and drawing the calibration curve. At the same time, the area under the curve (AUC) of the nomogram prediction model and the receiver operating characteristic (ROC) curve of the Leipzig score was calculated to compare the predictive ability of the nomogram model and the current Leipzig score for Wilson's disease. Results: Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), albumin (ALB), uric acid (UA), serum calcium (Ca), serum phosphorus (P), and hemoglobin (HGB) are closely related to the occurrence of Wilson's disease (p < 0.1). The prediction model of Wilson's disease contains seven independent predictors: ALT, AST, AKP, ALB, UA, Ca, and P. The AUC value of the prediction model was 0.971, and the C-index value was 0.972. The calibration curve was well fitted with the ideal curve. The nomogram prediction model had a good predictive effect on the occurrence of Wilson's disease; the ROC curve of Leipzig score was drawn, and the AUC value was calculated. The AUC of the Leipzig score was 0.969, indicating that the prediction model and the scoring system had predictive value, and the nomogram prediction model had a better predictive effect on the research objects of the center. Conclusion: ALT, AST, AKP, ALB, UA, Ca, and P are independent predictors of Wilson's disease, and can be used as early predictors. Based on the nomogram prediction model, the optimal threshold was determined to be 0.698, which was an important reference index for judging Wilson's disease. Compared with the Leipzig score, the nomogram prediction model has a relatively high sensitivity and specificity and has a good clinical application value.

Loureirin B inhibits the proliferation of hepatic stellate cells and the Wnt/ß-catenin signaling pathway by regulating miR-148-3p.

BACKGROUND: We investigated the activity of loureirin B against liver fibrosis and the underlying molecular mechanisms. METHODS: Hepatic stellate cells (HSCs) from Sprague-Dawley rats were treated with different concentrations of loureirin B. We used the MTT assay to determine HSC proliferation, flow cytometry to analyze apoptosis, and western blot to determine the expressions of Bax, Bcl-2, Wnt1 and ß-catenin. Real-time PCR was used to determine the expressions of Wnt1 and miR-148-3p. RESULTS: The MTT assay showed that loureirin B treatment significantly inhibited the proliferation of HSCs in time- and dose-dependent manners. Loureirin B significantly promoted the apoptosis of HSCs, increased the expression of Bax and decreased the Bcl-2 level. Western blot analysis showed that the expressions of Wnt1 and ß-catenin were obviously lower in the loureirin B treatment group than in the control group. We also found that loureirin B could decrease the Wnt1 mRNA level and increase miR-148-3p expression. Knockdown of miR-148-3p using inhibitor could reverse the effects of loureirin B on the proliferation and apoptosis of HSCs and the expressions of Bax, Bcl-2, Wnt1 and ß-catenin. CONCLUSION: Our results suggest that loureirin B inhibited the proliferation and promoted the apoptosis of HSCs, and suppressed the Wnt/ß-catenin signaling pathway via regulation of miR-148-3p.

Effect of different treatments and alcohol addiction on gut microbiota in minimal hepatic encephalopathy patients.

Minimal hepatic encephalopathy (MHE) is caused by dysbiosis of gut microbiota, particularly the ammonia-producing bacteria. Given the efficacy of certain treatments on MHE and the connection between alcoholism and MHE, a thorough understanding of how these strategies affect the gut microbiota in patients (alcoholic or non-alcoholic) will facilitate the assessment of their efficacy in the reshaping of gut microbiota. In the present study, a metagenomics approach was adopted to reveal alterations in gut microbiota of 14 MHE patients following treatment with rifaximin alone or rifaximin plus probiotics. Patients were grouped into the alcoholic and non-alcoholic groups to examine differences in terms of their response to treatment. Treatment reduced the overall microbiota diversity and decreased the abundance of certain ammonia-producing bacteria, such as Clostridium, with the treatment of rifaximin plus probiotics presenting a more apparent effect. Non-alcoholic MHE patients responded better to the treatment, as they presented greater reduction in microbiota diversity and a more consistent decline in certain ammonia-producing bacteria genera (such as Clostridium and Streptococcus) belonging to the Firmicutes phylum. In conclusion, treatment with rifaximin alone and rifaximin plus probiotics exhibited a different effect in different MHE patients, decreasing the overall gut microbiota diversity to various extents and reshaping microbiota in different ways. Furthermore, non-alcoholic MHE patients responded better to treatment in microbiota alterations.

[Gut flora and gut-derived endotoxin in minimal hepatic encephalopathy].

OBJECTIVE: To investigate the relationship of gut flora and gut-derived endotoxin with minimal hepatic encephalopathy (MHE). METHODS: Patients with hepatitis B virus-related liver cirrhosis (HBV-LC) were screened for MHE using the number connect test-A (NCT-A) and digital symbol test (DST) and divided into the following groups: HBV-LC with (+) MHE (n = 26) and HBV-liver cirrhosis without (-) MHE (n = 25); in addition, one healthy immediate family member of each patient in the HBV-LC + MHE group was enrolled as a control. Each participant provided fecal and blood samples. PCR amplification and 454 pyrosequencing were used to detect bacterial 16S rRNA in feces. Turbidimetric Limulus amebocyte lysate assay was used to detect level of endotoxin in serum. The significance of inter-group differences was assessed by one-way ANOVA or Student's t-test. RESULTS: The three groups showed different distributions of gut flora. The differences in the microbial communities' members and distributions were related to disease or health status, but not to the patient's genetic makeup or diet. In particular, the HBV-LC + MHE patients showed significantly lower amounts of different bacterial species and abundance of these species than the other two (non-MHE) groups (P less than 0.05). The healthy control family members had a richer diversity of gut flora than their counterparts with HBV-LC + MHE (P less than 0.05). The HBV-LV + MHE patients also had higher serum levels of endotoxin. CONCLUSION: Development of minimal hepatic encephalopathy in patients with HBV-LC may be related to a gut flora disorder or higher levels of endotoxin in serum.