Introduction Secreted phosphoprotein 1 (SPP1) is upregulated in cancers, but its role in metastatic colorectal cancer (CRC) and expression in HIV-associated CRC remain unclear.Methods Transcriptomic data from the GEO and TCGA databases were analyzed. Metastasis-specific genes were identified. SPP1-associated functions were explored using GO enrichment, KEGG pathway analysis, and GSVA. Prognostic value was assessed. SPP1 expression was evaluated in 30 CRC specimens (15 HIV-positive, 15 HIV-negative) via immunohistochemistry.Results SPP1 exhibited a metastasis-specific expression pattern. High SPP1 correlated with significantly elevated immune cell infiltration and adverse clinicopathological features: older age, mucinous adenocarcinoma, lymphatic invasion, advanced stage, high TN stage, MSI-H status, absence of polyps, and poor prognosis. GO enrichment linked SPP1 to extracellular matrix organization, cell adhesion, immune response, inflammation, and receptor binding. KEGG analysis showed enrichment in HIV-1 infection pathways. However, overall SPP1 expression did not significantly differ between HIV-positive and HIV-negative CRC tissues. Diffuse SPP1 protein expression was noted in an HIV-positive signet-ring cell carcinoma.Discussion SPP1 is a pivotal driver of CRC metastasis and immune regulation in the tumor microenvironment, and is associated with aggressive disease and poor outcomes. It shows strong potential as a prognostic biomarker. While HIV status did not broadly alter SPP1 expression, its specific pattern in certain HIV-associated carcinomas requires further study.Conclusion SPP1 critically mediates metastasis and immune regulation in CRC and independently predicts poor survival. Its prognostic value is confirmed, warranting investigation into its role in specific HIV-related carcinomas.

ABSTRACT Porcine contagious pleuropneumonia caused by Actinobacillus pleuropneumoniae (APP) is a fatal respiratory disease that threatens the worldwide farming industry’s health. The immune responses of extrapulmonary tissues play an important role in developing porcine contagious pleuropneumonia; however, the immune responses of extrapulmonary tissues induced by APP are rarely uncovered. Here, we used high-dimensional mass cytometry to investigate the immune cell response in the spleen and peripheral blood during APP infection in mice. We found that the immune response triggered by APP was highly tissue-specific. Numerous infection time- or tissue-specific immune cell clusters, including previously unrecognized ones, were also identified in the spleen and peripheral blood. Integrative analysis of splenic lymphoid and myeloid cell clusters maps the dynamic immune response cellular network during APP infection. Surprisingly, during the early stages of APP infection, the majority of the top 6 cell clusters contributing to the infection time-specificity in the spleen were adaptive immune cell clusters rather than innate immune cell clusters, among which CD24hiMHCII+CD8+TEM cells exhibited a stronger expression of IFN-γ, IL-17A, and IL-10 compared to the CD24lo compartment. In peripheral blood, there was unprecedented heterogeneity in the immune cell composition. Also, peripheral immune cell clusters closely related to the severity of APP infection were identified. In summary, our data provide a systemic and comprehensive overview of the immune responses to APP infection in the spleen and peripheral blood. This provides a foundation for understanding the immune pathogenesis of APP and identifying potential diagnostic biomarkers and therapeutic targets.IMPORTANCEThis study explored the cross-tissue immune dynamic landscape in the APP-induced pneumonia model by utilizing high-dimensional mass cytometry. We discovered that APP-induced immune responses are tissue-specific. Key infection-specific clusters in the spleen and peripheral blood were identified, some of which were previously unrecognized. Meanwhile, the specific functions of APP infection-related immune subsets were explored. The research systematically outlined an overview of immune responses in these tissues, deepening the understanding of APP pathogenesis and laying the foundation for the search for diagnostic and therapeutic targets.

The rapid expansion of next-generation sequencing (NGS) databases over the past decade has significantly advanced the identification of novel viruses across a wide range of host species. The Serratus platform and the NCBI Sequence Read Archive (SRA) database were utilized to reassess and analyze publicly available NGS datasets, aiming to identify novel hepadnaviruses and nackednaviruses. Our analysis uncovered multiple complete genomes of previously unrecognized hepadnaviruses and nackednaviruses, including those putatively infecting animals such as hamsters and buffaloes. Additionally, we identified the presence and distribution of various hepadnaviruses and nackednaviruses in African cichlid fishes. In vitro assays employing replication-competent plasmids derived from the identified rice rat and frog hepadnaviruses demonstrated their capacity to support viral replication. The identification of these novel hepadnavirus and nackednavirus species provides valuable insights into the origin and evolutionary history of hepadnaviruses. Moreover, these findings open new avenues for investigating potential animal models to study hepadnavirus replication and infection.

BACKGROUND:Low-level viremia (LLV) during antiviral therapy predicts poor outcomes in chronic hepatitis B (CHB). Although bile acids (BAs) regulate HBV transcription, their role in LLV remains unclear. This study aimed to determine whether specific BAs affect antiviral efficacy and to identify potential therapeutic targets. METHODS:We analyzed 111 CHB patients, including LLV and lower detection limit (LDL) groups. Serum BAs were profiled by mass spectrometry. Propensity score matching (PSM), correlation, and ROC analyses evaluated the association between BAs and HBV DNA. In vitro, deoxycholic acid (DCA) was tested in HBV-producing HepG2.2.15 and HBV-infected HepG2-NTCP cells. In vivo, HBV-transgenic (HBV-Tg) mice were used to assess hepatic DCA and HBV DNA correlation. Treatments included entecavir (ETV), antibiotics, and DCA supplementation. Molecular docking and mutagenesis were conducted to explore DCA-HBV surface protein (HBs) interactions. RESULTS:After PSM, LLV patients had elevated DCA, LCA, and TUDCA levels, which correlated with HBV DNA. DCA modestly predicted LLV by ROC analysis. In vitro, DCA promoted HBV Dane particle secretion and infection without altering viral protein expression. In HBV-Tg mice, hepatic DCA correlated positively with HBV DNA. ETV and antibiotics reduced DCA and HBV DNA, whereas DCA supplementation reversed these effects. Mechanistically, DCA bound to LXXLL motifs in the HBs transmembrane domains, particularly TM2. TM2 mutations disrupted DCA binding, HBs-HBc interaction, and Dane particle formation. CONCLUSION:Elevated DCA promotes viral persistence by enhancing Dane particle formation and stabilizing HBs-HBc interactions, contributing to LLV during treatment. Modulating BA metabolism may offer new strategies to improve CHB therapy.

In persons living with HIV (PWH), non-AIDS-related tumors, including colorectal cancer (CRC), have become major health concerns worldwide since the introduction of highly active antiretroviral therapy. To date, no study has addressed the underlying molecular mechanisms in PWH with CRC. To explore the impact of PWH with CRC, we sequenced total RNA and DNA from individuals with HIV-negative and PWH formalin-fixed paraffin-embedded (FFPE) CRC for transcriptome and genome analyses. We performed RNA and DNA extraction from FFPE samples, library preparation, total RNA sequencing, and whole-genome sequencing. A total of 1,705 genes were found to be differentially expressed genes (DEGs), including 1,121 upregulated DEGs and 584 downregulated DEGs, in PWH compared with HIV-negative CRC. Functional pathway analysis revealed that the DEGs were enriched mainly in infectious and immune diseases and various metabolic processes. The immune infiltration results revealed that the numbers of activated dendritic cells (aDCs), natural killer T cells (NKT cells), and T follicular helper cells (Tfh cells) were greater and that the number of memory B cells was lower in patients with CRC than in PWH. Twelve hub genes involved in interferon-stimulated genes (ISGs)-IFI44, MX1, OAS1, OAS3, BST2, IFIT1, FGF2, EGF, CCL3, CCL4, SHH, and PPARG-are positively related to aDC, NKT, Tfh, and memory B cells. We found highly analogous insertions, deletions, and functional annotations of the detected single nucleotide polymorphisms and indel mutations in PWH and patients with HIV-negative CRC. This study provides new insights into crucial ISGs, immune infiltration, immune variants, and pathways involved in CRC with HIV infection.

IL-21/IL-21R signaling is crucial in various immune diseases and cellular development, however, its role in bacterial pneumonia remains unclear. Here, IL-21R knockout (IL-21R−/−) mice were more susceptible to Actinobacillus pleuropneumoniae (APP) than wild-type (WT) mice. High-dimensional mass cytometry analysis revealed that IL-21R deficiency inhibited neutrophil activation, decreased the numbers of monocytes and proinflammatory macrophages, and augmented the defective CD3low T cells in the lungs. Intracellular cytokine staining showed decreased IFN-γ/TNF-α/IL-6 production in IL-21R−/− mice, particularly in CD8⁺ T cells. Furthermore, a previously unrecognized Ly6C+Ly6G+CD4+ T cell subset emerged only in the lungs of WT mice post-APP infection, which was in an activated status with stronger secretion capacities of IL-10, IL-21, granzyme B, and perforin by flow cytometry. These cells polarized macrophages into M2- or M1- phenotype without/with infection, respectively, and enhanced proliferation, phagocytosis, and macrophage extracellular traps/ROS-mediated bactericidal activity of macrophages against-APP, Klebsiella pneumoniae, or Escherichia coli infection. Thus, our study demonstrated that IL-21 drives the differentiation of neutrophils, monocytes, and macrophages into pro-inflammatory subsets. IL-21-induced Ly6C+Ly6G+CD4+ T cells cooperate with macrophages to enhance bacterial clearance, providing a promising target for preventing bacterial pneumonia.

Chronic hepatitis B (CHB), driven by persistent hepatitis B virus (HBV) infection, is characterised by unresolved liver inflammation and fibrosis. Despite its clinical burden, the immune mechanisms underpinning CHB progression, particularly the role of macrophage polarisation, remain incompletely defined. We integrated multi-modal approaches to dissect the CHB immune microenvironment: immunohistochemistry (HBsAg/HBcAg quantification), imaging mass cytometry (spatial immune mapping), microfluidic high-throughput qPCR (gene profiling) and MILLIPLEX assays (cytokine quantification). In vitro, HBV-producing HepG2.2.15 cells were cocultured with polarised THP-1 macrophages (M1/M2) and LX-2 hepatic stellate cells (HSCs) to model macrophage-HSCs crosstalk. CHB severity correlated with elevated virologic markers (HBsAg, HBeAg, HBV DNA) and liver injury indices (ALT/AST). The hepatic immune landscape was dominated by M1-like macrophages, which colocalised with activated HSCs and collagenⅠ+ fibrotic niches. Intrahepatic M1 markers (CD86, TNFα, CXCL9 and CXCR3) were upregulated, while the M2 marker IL-10 was suppressed. Serum HBeAg levels positively correlated with intrahepatic CD86 and CXCL9, implicating HBeAg as a key driver of M1 polarisation. Compartment-specific cytokine profiling revealed elevated liver-to-plasma ratios of TGF-α, IFN-γ and IP-10 in advanced CHB, contrasting with reduced IL-10. In vitro, HBV skewed THP-1 macrophages towards an M1 phenotype and HBV-primed M1 macrophages potently activated LX-2 cells. Persistent HBV infection fuels CHB progression by fostering a pro-inflammatory M1 macrophage-dominated microenvironment, which synergises with HSCs activation and fibrogenesis. Our findings nominate M1 polarisation as therapeutic targets to disrupt inflammation-fibrosis crosstalk in CHB.

BackgroundAcute liver injury (ALI), which is a type of inflammation-mediated hepatocellular injury, is a clinical syndrome that results from hepatocellular apoptosis and hemorrhagic necrosis. Apoptosis stimulating protein of p53-2 (ASPP2) is a proapoptotic member of the p53 binding protein family. However, the role of ASPP2 in the pathogenesis of ALI and its regulatory mechanisms remain unclear.MethodsThe expression of ASPP2 were compared between liver biopsies derived from patients with CHB, patients with ALI, and normal controls. Acute liver injury was modelled in mice by administration of D-GalN/LPS. Liver injury was demonstrated by serum transaminases and histological assessment of liver sections. ASPP2-knockdown mice (ASPP2+/−) were used to determine its role in acute liver injury. Mouse bone marrow macrophages (BMMs) were isolated from wildtype and ASPP2+/- mice and stimulated with LPS, and the supernatant was collected to incubate with the primary hepatocytes. Quantitative real-time PCR and western blot were used to analyze the expression level of target.ResultsThe expression of ASPP2 was significantly upregulated in the liver tissue of ALI patients and acute liver injury mice. ASPP2+/- mice significantly relieved liver injury through reducing liver inflammation and decreasing hepatocyte apoptosis. Moreover, the conditioned medium (CM) of ASPP2+/- bone marrow-derived macrophages (BMMs) protected hepatocytes against apoptosis. Mechanistically, we revealed that ASPP2 deficiency in BMMs specifically upregulated IL-6 through autophagy activation, which decreased the level of TNF-α to reduce hepatocytes apoptosis. Furthermore, up-regulation of ASPP2 sensitizes hepatocytes to TNF-α-induced apoptosis.ConclusionOur novel findings show the critical role of ASPP2 in inflammatory immunoregulatory mechanism of ALI and provide a rationale to target ASPP2 as a refined therapeutic strategy to ameliorate acute liver injury.

BACKGROUND:Primary biliary cholangitis (PBC) is associated closely with the gut microbiota. This study aimed to explore the characteristics of the gut microbiota after the progress of PBC to cirrhosis. METHOD:This study focuses on utilizing the 16S rRNA gene sequencing method to screen for differences in gut microbiota in PBC patients who progress to cirrhosis. Then, we divided the data into training and verification sets and used seven different machine learning (ML) models to validate them respectively, calculating and comparing the accuracy, F1 score, precision, and recall, and screening the dominant intestinal flora affecting PBC cirrhosis. RESULT:PBC cirrhosis patients showed decreased diversity and richness of gut microbiota. Additionally, there are alterations in the composition of gut microbiota in PBC cirrhosis patients. The abundance of Faecalibacterium and Gemmiger bacteria significantly decreases, while the abundance of Veillonella and Streptococcus significantly increases. Furthermore, machine learning methods identify Streptococcus and Gemmiger as the predominant gut microbiota in PBC patients with cirrhosis, serving as non-invasive biomarkers (AUC = 0.902). CONCLUSION:Our study revealed that PBC cirrhosis patients gut microbiota composition and function have significantly changed. Streptococcus and Gemmiger may become a non-invasive biomarker for predicting the progression of PBC progress to cirrhosis.

Primary biliary cholangitis (PBC) is an autoimmune liver disease. During the diagnostic process, the patient's autoimmune antibodies are routinely examined. Approximately 20% of PBC patients have positive anti-centromere antibody (ACA). We evaluated the clinical characteristics of ACA-positive and ACA-negative PBC patients to explain the differences in disease progression between these two groups. Retrospective data from 961 PBC patients at Beijing Youan Hospital from 2010 to 2019 were gathered and separated into two groups based on ACA positivity. We collected and evaluated clinical laboratory indices, gastroscopy findings, and liver function assessments. In addition, 60 liver biopsies were available for comparison between the 2 groups. Pathologists staged the histological findings using the Ludwig staging criteria and Nakanuma staging and grading. Immunohistochemical staining was also performed on liver biopsies to examine the expression of cytokeratin 7 (CK7) in the tissue. A synthesis of clinical indicators in the large cohort showed that alanine transaminase, aspartate aminotransferase, total bilirubin, IgG, white blood cell, and platelet were significantly lower in the ACA-positive group, indicating that the overall status of liver injury was more moderate in the ACA-positive group. Additionally, ACA-positive patients in the non-cirrhotic group were more likely to present with gastroesophageal varices related to portal hypertension. Finally, analysis of pathologic findings showed that parameters were mostly comparable in the two groups, but CK7 differed and was more significantly lower in the ACA-positive group in albumin-bilirubin grade 2 and 3 patients. In summary, we characterized and compared the clinical features of ACA-positive and ACA-negative PBC patients, corroborating previous studies on the relationship between ACA positivity and portal hypertension cross-sectionally. It suggested that gastroesophageal varices might happen in the earlier course of PBC natural progression in the ACA-positive group.

BackgroundThe mechanism of decreased T cells infiltrating tumor tissues in hepatocellular carcinoma is poorly understood.MethodsCells were separated from the single-cell RNA-sequence dataset of hepatocellular carcinoma patients (GSE149614) for cell-cell communication. Flow cytometry, EDU staining, H3-Ser28 staining, confocal immunofluorescence staining, western blotting and naked microsubcutaneous tumors were performed for the mechanism of NGF-NGFR promoting proliferation.ResultsThe present study has revealed that during the process of T-cell infiltration from adjacent tissues to tumor tissues, an inefficiency in NGF-NGFR communication occurs in the tumor tissues. Importantly, NGF secreted by tumor cells interacts with NGFR present on the membranes of the infiltrated T cells, thereby promoting the proliferation through the activation of mitotic spindle signals. Mechanistically, the mediation of mitotic spindle signal activation promoting proliferation is executed by HDAC1-mediated inhibition of unclear trans-localization of PREX1. Furthermore, PD-1 mAb acts synergistically with the NGF-NGFR communication to suppress tumor progression in both mouse models and HCC patients. Additionally, NGF-NGFR communication was positively correlates with the PD-1/PDL-1 expression. However, expressions of NGF and NGFR are low in tumor tissues, which is responsible for the invasive clinicopathological features and the disappointing prognosis in HCC patients.ConclusionInefficiency in NGF-NGFR communication impairs PD-1 mAb immunotherapy and could thus be utilized as a novel therapeutic target in the treatment of HCC patients in clinical practice.

Non-alcoholic fatty liver disease (NAFLD) affects approximately 25 % of the global population, presenting a significant threat to human health and imposing a substantial economic burden on society. Cytokeratin 18 (CK18) has emerged as a promising non-invasive biomarker for early detection of NAFLD. However, current diagnostic methods face challenges in accurately identifying NAFLD in its early stages. To address this issue, our study aimed to generate new CK18 antibodies and utilize a highly sensitive single -molecule immunodetection system customized for NAFLD. We embarked on a process involving the production of novel CK18 monoclonal antibodies, which were subsequently engineered into recombinant forms. In comparison to commercial antibodies, our antibodies exhibited superior affinity. Grounded in the SMCxPRO platform, our assay underwent rigorous assessment, adhering to well-defined biomarker assay criteria. The assay demonstrated specificity to CK18 and achieved a lower limit of quantification of 1 pg/ml. Importantly, elevated serum CK18 levels were observed among NAFLD patients compared to healthy individuals. The single -molecule detection system, utilizing our novel antibodies, holds significant promise for the early -stage diagnosis, dynamic disease monitoring, and prognostic evaluation of NAFLD patients.

Hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), has a significantly higher risk of recurrence. However, the exact mechanism by which HBV prompts HCC recurrence remains largely unknown. In this study liver microarray test revealed significant upregulation of microtubule associated protein 1S (MAP1S) in metastatic HCC compared to control. MAP1S knockdown suppressed growth of HCCLM3 cells in vitro and in vivo. Mechanistically, HBV-encoded X protein (HBx) upregulates MAP1S, which enhances microtubule (MT) acetylation by promoting the degradation of histone deacetylase 6 (HDAC6), and facilitates the nuclear translocation of Smad complex, and thereby enhancing downstream TGF-β signaling. Smad complex, in turn, increases MAP1S, establishing a feedback loop of MAP1S/Smad/TGF-β1. Finally, survival analysis of 150 HBV-associated HCC patients demonstrated both increased MAP1S and decreased HDAC6 were significantly associated with shorter relapse-free survival. Collectively, this study reveals a unique mechanism whereby HBx-induced upregulation of MAP1S drives HBV-related HCC proliferation and migration through the MAP1S/Smad/TGF-β1 feedback loop. TEASER: MAP1S is a key link between HBV infection and a higher risk of metastatic recurrence of HCC.

BackgroundRadiofrequency ablation (RFA) is the primary curative treatment for hepatocellular carcinoma (HCC) patients who are not eligible for surgery. However, the effects of RFA on the global tumor immune response remain unclear.MethodIn this study, we examined the phenotypic and functional changes in peripheral blood mononuclear cells (PBMCs) from recurrent HCC patients who had undergone two RFA treatments using mass cytometry and high-throughput mRNA assays. ResultsWe observed significant increase in monocytes and decrease in T cell subpopulations three days after the first RFA treatment and three days after the second RFA treatment. The down-regulation of GZMB, GZMH, GZMK, and CD8A, which are involved in the cytotoxic function of T cells, was observed following RFA. Furthermore, the population of CD8 effector and memory T cells (CD8 Teff and CD8 Tem) significantly decreased after RFA. The expression of CD5 and CD161 in various T cell subpopulations also showed significant reductions. Additionally, elevated secretion of VEGF was observed in monocytes, B cells, regulatory T cells (Tregs), and CD4 naive T cells. ConclusionIn recurrent HCC patients, serum components derived from radiofrequency therapy can enhance the antigen-presenting capacity of monocytes. However, they also inhibit the anti-cancer immune response by reducing the population of CD8 effector and memory T cells and suppressing the activation of T cells, as well as down-regulating the expression of CD161 and CD5 in various T cell subpopulations. These tumor-derived components also contribute to an immunosuppressive microenvironment by promoting the secretion of VEGF in monocytes, Tregs, B cells, and CD4 naive T cells.

The initial stage of alcoholic liver disease (ALD) is hepatic steatosis. Recent studies have highlighted a possible role for Apoptosis-stimulating protein 2 of p53 (ASPP2) in regulating hepatic lipid metabolism in nonalcoholic fatty liver (NAFLD). However, whether ASPP2 regulates alcohol-induced lipid accumulation and its mechanisms remain unclear. To explore that, we establish an alcoholic liver injury model in vivo and in vitro. The clinical specimens were collected from liver tissues of patients with alcoholic liver disease. Lipid metabolism was detected by HE staining, oil red O staining and qPCR; and ASPP2-peroxisome proliferator-activated receptor γ (PPARγ) signaling pathways were detected by western blot and immunohistochemical staining. We found that both ASPP2 and PPARγ expression increased in patients and mouse models with ALD. We also discovered the reduction of ASPP2 significantly inhibited the expression of PPARγ and alleviated alcohol-induced hepatic lipid accumulation and liver injury in vivo and in vitro. Mechanistically, the PPARγ agonist reversed the protective effect of ASPP2 downregulation on hepatic steatosis and liver injury, while the opposite results were observed using PPARγ inhibitor. In conclusion, ASPP2 exacerbates ethanol-induced lipid accumulation and hepatic injury by upregulating the PPARγ signaling pathway, thus promoting the occurrence and development of ALD.

在新的教学形势下,传染病学教研室开展传染病学课程的线上教学,使用钉钉+雨课堂+腾讯会议为主要教学软件对课前、课中、课后三个教学环节进行探索,总结分析教学过程中各软件优势与弊端.实践表明,传染病学教学中,多种线上授课软件的相互结合能达到甚至超过传统授课的效果,对于新形势下医学人才的培养具有重要的意义.

Objective The purpose of this study was to establish a rapid and efficient liquid chromatography tandem mass spectrometry（LC-MS/MS） for simultaneous determination of 15 bile acids in mouse liver tissues. Methods The activated charcoal was utilized to prepare bile acid-free liver, which served as the biological matrix for the preparation of standard and quality control samples. The mouse liver tissue was homogenized, and a basic acetonitrile solution, including 5% NH 4 OH was added to precipitate proteins. The proteins were separated on an Agilent Poroshell 120 EC C18 column（100 mm×4.6 mm, 2.7 μm） by using ~2H 4 -DCA, GUDCA-d 5 , and LCA-d 4 as internal standards. The mobile phase is ammonium acetate aqueous solution and methanol acetonitrile mixed solution for gradient elution, the column temperature was 30℃, the flow rate was 0.3mL/min, and the injection volume was 2 μL. The electrospray ion source（ESI） was operated in negative ion mode, and in multiple reaction monitoring（MRM）. Results The linearity of the 15 bile acids was good with R 2 greater than 0.993, the limits of determination were less than 2 ng/mL, and the matrix effects were 90.76%-109.25%; the intra-day and inter-day accuracy and precision were less than 15%, and the stability was good under 4℃ for 24 h, repeated freeze-thaw, and freeze-storage for one month, meeting the analytical requirements of biological samples; the detection of mouse liver tissues showed that both unconjugated BAs and conjugated BAs（G-BAs, T-BAs） were dominated by maternal CA, with the highest content of TCA; the concentration of unconjugated BAs was（723.89±50.65） ng/mL, significantly higher than that of G-BAs [（56.90±11.28） ng/mL, P＜0.001]; the concentration of T-BAs was（40322.90±14034.80）ng/mL, significantly higher than unconjugated BAs（P＜0.001）, and also significantly higher than G-BAs（P＜0.001）. Conclusion The LC-MS/MS method we established is sensitive, accurate, reliable, and suitable for the determination of bile acids concentrations in mouse liver tissues, which might help for further studies.

Hepatocellular carcinoma（HCC） is the most prevalent form of primary liver cancer, accounting for 75%-85% of cases. Although treatments are given to cure early-stage HCC, up to 50%-70% of individuals may experience a relapse of the illness in the liver after 5 years. Research on the fundamental treatment modalities for recurrent HCC is moving significantly further. The precise selection of individuals for therapy strategies with established survival advantages is crucial to ensuring better outcomes. These strategies aim to minimize substantial morbidity,support good life quality, and enhance survival for patients with recurrent HCC.For individuals with recurring HCC after curative treatment, no approved therapeutic regimen is currently available. A recent study presented novel approaches, like immunotherapy and antiviral medication, to improve the prognosis of patients with recurring HCC with the apparent lack of data to guide the clinical treatment. The data supporting several neoadjuvant and adjuvant therapies for patients with recurring HCC are outlined in this review. We also discuss the potential for future clinical and translational investigations.

Objective: To screen and identify differential proteins, analyze lipid metabolism-related proteins and pathways, and explore their functions and biological processes in liver tissue of patients with alcoholic liver disease using tandem mass tag (TMT) labeling technology. Methods: Liver tissues that met the inclusion criteria were collected. Eight samples from patients with alcoholic cirrhosis and three samples from the normal control group were screened out. The TMT technique was used to screen differential proteins, perform signaling pathway enrichment analysis, and analyze protein interaction networks to explore the biological processes involved in them. Results: Proteomic analysis identified 2 741 kinds of differentially expressed proteins in the two groups of data with statistical significance (P < 0.05). The standard criteria of P < 0.05 and |log2(foldchange)| > 1 had screened out 106 kinds of differentially expressed proteins. Compared with the control group, the alcoholic liver disease group had 12 kinds of up-regulated proteins and 94 kinds of down-regulated proteins. Among them, there were 2 kinds of up-regulated differential proteins related to lipid metabolism and 14 kinds of down-regulated differential proteins. The results of bioinformatics analysis showed that these proteins were primarily involved in biological processes such as lipid transport, regulation of lipase activity, fatty acid binding, and cholesterol metabolism in lipid metabolism and also had a close link to signal pathways related to lipid metabolism such as peroxisome proliferator-activated receptor signaling pathways, cholesterol metabolism, triglyceride metabolism, and regulation of lipolysis in adipocytes. Conclusion: The 16 kinds of lipid metabolism-related differential proteins may be the key proteins in the pathogenesis of alcoholic liver disease.

Background: With continuous advances in next-generation sequencing (NGS), novel hepadnaviruses have been discovered in many species over the last 10 years. Methods: In this study, the cloud search and analysis of NGS data published in Nature by Edgar et al. was used as a basis to re-mine and reanalyze public NGS data for new hepadnaviruses. Results: Ultimately, at least 41 new species of hepadnaviruses were identified, including hepadnaviruses from the model animals hamster and mouse, frog hepadnavirus with pan-species infectivity, and diverse African cichlid hepadnaviruses that circulate within populations. Conclusions: The discovery of the new species of hepadnaviruses not only provides new clues for the study of the origin and evolution of hepadnaviruses, but also can be used to construct new hepadnavirus animal infection models, which will be helpful for the research of eradicative drugs for hepatitis B virus (HBV).