Internet Gaming Disorder (IGD) is a recognized public health concern, particularly among young adults. Medical students in vocational colleges in China face a unique combination of high academic pressure and potential exposure to problematic gaming behaviors. This study aimed to determine the prevalence of IGD and investigate its relationship with key psychological correlates, including depression, anxiety, and mental well-being, in this specific population. This cross-sectional survey was conducted from January 2021 to March 2025 in four vocational colleges in Sichuan Province, China. A total of 7,045 medical undergraduates provided valid survey responses, exceeding the calculated minimum sample size of 6,793. Participants completed standardized instruments, including the Internet Gaming Disorder Scale-Short Form (IGDS9-SF), the Patient Health Questionnaire 9 (PHQ-9), the Generalized Anxiety Disorder 7-item (GAD-7), and the Warwick-Edinburgh Mental Well-being Scale (WEMWBS). Rigorous quality assurance was maintained through standardized participant training and data validation protocols, including a 5

Intrahepatic cholangiocarcinoma (ICC) is the second most prevalent type of primary liver cancer and lacks effective targeted therapy. Previously, we reported that B cell-specific Moloney murine leukaemia virus integration site 1 (Bmi1) drives the formation and development of ICC independent of Ink4A/Arf; however, the underlying mechanism remains unclear. Here, we report that hepatic leukaemia factor (HLF) acts as a tumour-suppressor gene in ICC and Bmi1 represses HLF to drive ICC initiation and progression. RNA sequencing was performed to find the downstream target of Bmi1 in ICC. Bioinformatic analysis and molecular biological techniques were used to examine the expression of Bmi1 and HLF in ICC. Effects of HLF silence or overexpression by lentivirus on cell proliferation or development were evaluated in human ICC cells, xenograft and primary ICC mouse models, respectively. The luciferase reporter assay was used to identify that Bmi1 regulates HLF. In ICC, HLF expression levels were inversely correlated with Bmi1. Overexpression of HLF inhibited the growth of ICC both in vitro and in vivo, whereas HLF knockout promoted ICC development in ICC mouse models. Importantly, HLF repression reversed the inhibitory effects of Bmi1 knockdown on cell survival, proliferation and colony formation. Luciferase reporter assay results indicated that Bmi1 represses HLF by directly binding to its promoter. These findings revealed the molecular mechanism through which Bmi1 promotes ICC formation and development and uncovered the role of HLF as a tumour suppressor in ICC.

Background and Aims:The causal biomarkers for metabolic dysfunction-associated steatotic liver disease (MASLD) and their clinical value remain unclear. In this study, we aimed to identify biomarkers for MASLD and evaluate their diagnostic and prognostic significance. Methods:We conducted a Mendelian randomization analysis to assess the causal effects of 2,925 molecular biomarkers (from proteomics data) and 35 clinical biomarkers on MASLD. Mediation analysis was performed to determine whether clinical biomarkers mediated the effects of molecular biomarkers. The association between key clinical biomarkers and MASLD was externally validated in a hospital-based cohort (n = 415). A machine learning-based diagnostic model for MASLD was developed and validated using the identified molecular biomarkers. Prognostic significance was evaluated for both molecular and clinical biomarkers. Results:Six molecular biomarkers-including canopy FGF signaling regulator 4 (CNPY4), ectonucleoside triphosphate diphosphohydrolase 6 (ENTPD6), and major histocompatibility complex, class I, A (HLA-A)-and eight clinical biomarkers (e.g., serum total protein (STP)) were identified as causally related to MASLD. STP partially mediated the effect of HLA-A on MASLD (23.61%) and was associated with MASLD in the external cohort (odds ratio = 1.080, 95% confidence interval: 1.011-1.155). A random forest model demonstrated high diagnostic performance (AUC = 0.941 in training; 0.875 in validation). High expression levels of CNPY4 and ENTPD6 were associated with the development of and poorer survival from hepatocellular carcinoma. Low STP (<60 g/L) predicted all-cause mortality (HR = 2.50, 95% confidence interval: 1.22-5.09). Conclusions:This study identifies six causal molecular biomarkers (e.g., CNPY4, ENTPD6, HLA-A) and eight clinical biomarkers for MASLD. Notably, STP mediates the effect of HLA-A on MASLD and is associated with all-cause mortality.

Objectives Primary liver cancer, including hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC), is a common malignancy lacking effective chemotherapies. Camptothecin (CPT) derivatives, though approved for treatment, face limitations due to systemic toxicity. Boration has been shown to improve drug bioavailability and reduce toxicity. We designed and synthesized a new 9-borate CPT derivative, CPT-01, to evaluate its anti-tumor potential. Methods In vitro assays, including EdU incorporation, colony formation, and flow cytometry, assessed CPT-01's effects on proliferation, apoptosis, and cell cycle. AKT/Ras- and AKT/NICD-induced primary mouse models of HCC and ICC were used for in vivo evaluation. Molecular mechanisms were explored using western blotting, immunohistochemistry, topoisomerase I inhibition assays, and molecular docking. Results CPT-01 exhibited stronger anti-tumor activity than topotecan (TPT) in both cancer cell lines and mouse models. In vivo, it showed greater efficacy in AKT/Ras-induced HCC and AKT/NICD-induced ICC models, with no significant toxicity at therapeutic doses. Mechanistically, CPT-01 inhibited topoisomerase I, inducing DNA damage, cell cycle arrest, and apoptosis. Conclusions CPT-01 demonstrates enhanced anti-tumor efficacy and reduced toxicity compared to TPT, offering promising clinical potential for liver cancer treatment.

BACKGROUND:This study utilised the Global Burden of Disease data (2010-2021) to analyse the rates and trends in point prevalence, annual incidence and years lived with disability (YLDs) for major chronic liver diseases, such as hepatitis B, hepatitis C, metabolic dysfunction-associated liver disease, cirrhosis and other chronic liver diseases. METHODS:Age-standardised rates per 100,000 population for prevalence, annual incidence and YLDs were compared across regions and countries, as well as the socio-demographic index (SDI). Trends were expressed as percentage changes (PC) and estimates were reported with uncertainty intervals (UI). RESULTS:Globally, in 2021, the age-standardised rates per 100,000 population for the prevalence of hepatitis B, hepatitis C, MASLD and cirrhosis and other chronic liver diseases were 3583.6 (95%UI 3293.6-3887.7), 1717.8 (1385.5-2075.3), 15018.1 (13756.5-16361.4) and 20302.6 (18845.2-21791.9) respectively. From 2010 to 2021, the PC in age-standardised prevalence rates were-20.4% for hepatitis B, -5.1% for hepatitis C, +11.2% for MASLD and + 2.6% for cirrhosis and other chronic liver diseases. Over the same period, the PC in age-standardized incidence rates were -24.7%, -6.8%, +3.2%, and +3.0%, respectively. Generally, negative associations, but with fluctuations, were found between age-standardised prevalence rates for hepatitis B, hepatitis C, cirrhosis and other chronic liver diseases and the SDI at a global level. However, MASLD prevalence peaked at moderate SDI levels. CONCLUSIONS:The global burden of chronic liver diseases remains substantial. Hepatitis B and C have decreased in prevalence and incidence in the last decade, while MASLD, cirrhosis and other chronic liver diseases have increased, necessitating targeted public health strategies and resource allocation.

BACKGROUND & AIMS: Metabolic dysfunction-associated steatohepatitis (MASH) and fibrotic MASH are significant health challenges. This multi-national study aimed to validate the acMASH index (including serum creatinine and aspartate aminotransferase concentrations) for MASH diagnosis and develop a new index (acFibroMASH) for non-invasively identifying fibrotic MASH and exploring its predictive value for liver-related events (LREs). METHODS: We analyzed data from 3004 individuals with biopsy-proven metabolic dysfunction-associated steatotic liver disease (MASLD) across 29 Chinese and 9 international cohorts to validate the acMASH index and develop the acFibroMASH index. Additionally, we utilized the independent external data from a multi-national cohort of 9034 patients with MASLD to examine associations between the acFibroMASH index and the risk of LREs. RESULTS: In the pooled global cohort, the acMASH index identified MASH with an area under the receiver operating characteristic curve (AUROC) of 0.802 (95% confidence interval [CI], 0.786-0.818). The acFibroMASH index (including the acMASH index plus liver stiffness measurement) accurately identified fibrotic MASH with an AUROC of 0.808 in the derivation cohort and 0.800 in the validation cohort. Notably, the AUROC for the acFibroMASH index was 0.835 (95% CI, 0.786-0.882), superior to that of the FAST score at 0.750 (95% CI, 0.693-0.800; P < .01) in predicting the 5-year risk of LREs. Patients with acFibroMASH >0.39 had a higher risk of LREs than those with acFibroMASH <0.15 (adjusted hazard ratio, 11.23; 95% CI, 3.98-31.66). CONCLUSIONS: This multi-ethnic study validates the acMASH index as a reliable, noninvasive test for identifying MASH. The newly proposed acFibroMASH index is a reliable test for identifying fibrotic MASH and predicting the risk of LREs.

Background & Aims:This study used the Global Burden of Disease data (2010-2021) to analyze the rates and trends of point prevalence, annual incidence, and years lived with disability (YLDs) for metabolic dysfunction-associated steatotic liver disease (MASLD) in 204 countries. Methods:Total numbers and age-standardized rates per 100,000 population for MASLD prevalence, annual incidence, and YLDs were compared across regions and countries by age, sex, and sociodemographic index (SDI). Smoothing spline models were used to evaluate the relationship between the burden of MASLD and SDI. Estimates were reported with uncertainty intervals (UI). Results:Globally, in 2021, the age-standardized rates per 100,000 population of point prevalence of MASLD were 15,018.1 cases (95% UI 13,756.5-16,361.4), annual incidence rates were 608.5 cases (598.8-617.7), and YLDs were 0.5 (0.3-0.8) years. MASLD point prevalence was higher in men than women (15,731.4 vs. 14,310.6 cases per 100,000 population). Prevalence peaked at ages 45-49 for men and 50-54 for women. Kuwait (32,312.2 cases per 100,000 people; 95% UI: 29,947.1-34,839.0), Egypt (31,668.8 cases per 100,000 people; 95% UI: 29,272.5-34,224.7), and Qatar (31,327.5 cases per 100,000 people; 95% UI: 29,078.5-33,790.9) had the highest prevalence rates in 2021. The largest increases in age-standardized point prevalence estimates from 2010 to 2021 were in China (16.9%, 95% UI 14.7%-18.9%), Sudan (13.3%, 95% UI 9.8%-16.7%) and India (13.2%, 95% UI 12.0%-14.4%). MASLD incidence varied with SDI, peaking at moderate SDI levels. Conclusions:MASLD is a global health concern, with the highest prevalence reported in Kuwait, Egypt, and Qatar. Raising awareness about risk factors and prevention is essential in every country, especially in China, Sudan and India, where disease incidence and prevalence are rapidly increasing. Impact and implications:This research provides a comprehensive analysis of the global burden of MASLD, highlighting its rising prevalence and incidence, particularly in countries with varying sociodemographic indices. The findings are significant for both clinicians and policymakers, as they offer critical insights into the regional disparities in MASLD burden, which can inform targeted prevention and intervention strategies. However, the study's reliance on modeling and available data suggests cautious interpretation, and further research is needed to validate these findings in clinical and real-world settings.

AIM:Non-invasive diagnostics for metabolic dysfunction-associated fatty liver disease (MAFLD) remain challenging. We aimed to identify novel key genes as non-invasive biomarkers for MAFLD, elucidate causal relationships between biomarkers and MAFLD and determine the role of immune cells as potential mediators. MATERIALS AND METHODS:Utilizing published transcriptome data of patients with biopsy-proven MAFLD, we applied linear models for microarray data, least absolute shrinkage and selector operation (LASSO) regressions and receiver operating characteristic (ROC) curve analyses to identify and validate biomarkers for MAFLD. Using the expression quantitative trait loci database and a cohort of 778 614 Europeans, we used Mendelian randomization to analyse the causal relationships between key biomarkers and MAFLD. Additionally, mediation analysis was performed to examine the involvement of 731 immunophenotypes in these relationships. RESULTS:We identified 31 differentially expressed genes, and LASSO regression showed three hub genes, IGFBP2, PEG10, and P4HA1, with area under the receiver operating characteristic (AUROC) curve of 0.807, 0.772 and 0.791, respectively, for identifying MAFLD. The model of these three genes had an AUROC of 0.959 and 0.800 in the development and validation data sets, respectively. This model was also validated using serum-based enzyme-linked immunosorbent assay data from MAFLD patients and control subjects (AUROC: 0.819, 95% confidence interval: 0.736-0.902). PEG10 was associated with an increased MAFLD risk (odds ratio = 1.106, p = 0.032) via inverse variance-weighted analysis, and about 30% of this risk was mediated by the percentage of CD11c + CD62L- monocytes. CONCLUSIONS:The MAFLD panels have good diagnostic accuracy, and the causal link between PEG10 and MAFLD was mediated by the percentage of CD11c + CD62L- monocytes.

Multidrug-resistant (MDR) Acinetobacter baumannii infections pose a significant challenge in burn wound management, necessitating the development of innovative therapeutic strategies. In this work, we introduced a novel polymyxin B (PMB)-targeted liposomal photosensitizer, HMME@Lipo-PMB, for precise and potent antimicrobial photodynamic therapy (aPDT) against burn infections induced by MDR A. baumanni. HMME@Lipo-PMB-mediated aPDT exhibited enhanced antibacterial efficacy by specifically targeting and disrupting bacterial cell membranes, and generating increased intracellular ROS. Remarkably, even at low concentrations, this targeted approach significantly reduced bacterial viability in vitro and completely eradicated burn infections induced by MDR A. baumannii in vivo. Additionally, HMME@Lipo-PMB-mediated aPDT facilitated burn infection wound healing by modulating M1/M2 macrophage polarization. It also effectively promoted acute inflammation in the early stage, while attenuated chronic inflammation in the later stage of wound healing. This dynamic modulation promoted the formation of granulation tissue, angiogenesis, and collagen regeneration. These findings demonstrate the tremendous potential of HMME@Lipo-PMB-mediated aPDT as a promising alternative for the treatment of burn infections caused by MDR A. baumannii.

BACKGROUNDLymphopenia and high CT score is associated with COVID-19 severity. Herein we describe the change pattern in lymphocyte count and CT score during hospitalization and explore a possible association with the severity of COVID-19.METHODSIn this retrospective study, 13 non-severe COVID-19 patients diagnosed at admission were enrolled. One patient progressed to severe disease. Change patterns in lymphocyte counts and CT scores of all patients were analyzed.RESULTSLymphocyte count increased gradually from day 5 post-illness onset (day 5 vs. day 15, p = 0.001). Lymphocyte count of the severe patient fluctuated at low levels throughout the 15-day period. Chest CT scores of non-severe patients increased significantly during the first 5 days of illness onset, but decreased gradually beginning day 9 (illness onset vs. day 5, p = 0.002, day 9 vs. day 15, p = 0.015). In the severe patient, CT score continued to increase over the 11 days post-illness onset period.CONCLUSIONSNon-severe COVID-19 patients had significantly increased lymphocyte counts and decreased CT scores beginning day 5 and day 9 of illness onset, respectively. The patients without increased lymphocyte counts and decreased CT scores during the early 2nd week of illness onset may develop to severe COVID-19.

BACKGROUND & AIMS:There is an unmet clinical need for non-invasive tests to diagnose non-alcoholic fatty liver disease (NAFLD) and individual fibrosis stages. We aimed to test whether urine protein panels could be used to identify NAFLD, NAFLD with fibrosis (stage F ≥ 1) and NAFLD with significant fibrosis (stage F ≥ 2).METHODS:We collected urine samples from 100 patients with biopsy-confirmed NAFLD and 40 healthy volunteers, and proteomics and bioinformatics analyses were performed in this derivation cohort. Diagnostic models were developed for detecting NAFLD (UPNAFLD model), NAFLD with fibrosis (UPfibrosis model), or NAFLD with significant fibrosis (UPsignificant fibrosis model). Subsequently, the derivation cohort was divided into training and testing sets to evaluate the efficacy of these diagnostic models. Finally, in a separate independent validation cohort of 100 patients with biopsy-confirmed NAFLD and 45 healthy controls, urinary enzyme-linked immunosorbent assay analyses were undertaken to validate the accuracy of these new diagnostic models.RESULTS:The UPfibrosis model and the UPsignificant fibrosis model showed an AUROC of .863 (95% CI: .725-1.000) and 0.858 (95% CI: .712-1.000) in the training set; and .837 (95% CI: .711-.963) and .916 (95% CI: .825-1.000) in the testing set respectively. The UPNAFLD model showed an excellent diagnostic performance and the area under the receiver operator characteristic curve (AUROC) exceeded .90 in the derivation cohort. In the independent validation cohort, the AUROC for all three of the above diagnostic models exceeded .80.CONCLUSIONS:Our newly developed models constructed from urine protein biomarkers have good accuracy for non-invasively diagnosing liver fibrosis in NAFLD.

Objective: To screen out potential prognostic hub genes for adult patients with sepsis via construction of a microRNA-mRNA-PPI network, and investigate the localization of these core genes in peripheral blood monocytes. Methods: The peripheral bloods of 33 subjects were performed microRNA and mRNA sequencing using high-throughput sequencing, differentially expressed genes and microRNAs were identified by bioinformatics. 10×single-cell transcriptome sequencing was conducted further. Results: Of which 23 adult septic patients and 10 healthy individuals, a relative levels of transcription of 20391 genes, and 1633 microRNAs were detected by RNA sequencing. A total of 1114 differentially expressed genes and 76 differentially expressed microRNAs were obtained using DESeq2, 454 differential expression genes (DEGs) were distinguished ultimately. The microRNA-mRNA-PPI network was constructed based on DEGs and the top 20 differentially expressed microRNAs, which included 10 up-regulated and 10 down-regulated microRNAs. Furthermore, hub genes TLR5, FCGR1A, ELANE, GNLY, IL2RB and TGFBR3 which may be association with the prognosis of sepsis and their negative correlation microRNAs were recognized. Genes TLR5, FCGR1A and ELANE were mainly expressed in macrophages, genes GNLY, IL2RB and TGFBR3 were expressed specifically in T cells and nature kill cells. Conclusions: The parallel analysis of mRNA and microRNA in patients with sepsis using RNA-seq was feasible. Potential hub genes and microRNAs which are possibly related to sepsis prognosis were identified, it provides new prospects for sepsis. However, further experiments need to be required.

Resveratrol (RES), a natural polyphenol phytoalexin, has been reported to attenuate nonalcoholic fatty liver disease (NAFLD). However, its roles on protection of liver from lipotoxicity and underlying mechanism are not fully understood. In this study, we investigated the impacts of RES on alleviating hepatic lipotoxicity and corresponding molecular mechanism. Impacts of RES on oleic acid (OA)-induced lipotoxicity were assessed in L02 cells and C57BL/6J mice, respectively. In L02 cells, lipotoxicity was assessed by detection of apoptosis, mitochondrial function, oxidative stress and ROS-related signaling. In mice, lipotoxicity was evaluated by detecting hepatic function, serum enzyme activity, and reactive oxygen species (ROS) levels. We found that RES reduced OA-induced apoptosis, mitochondrial dysfunction, ROS generation, and DNA damage in L02 cells. RES also decreased expression of cleaved caspase-3 and p53 and increased expression of B-cell lymphoma 2 (Bcl-2). Importantly, RES protected mice from high-fat diet-induced hepatic lipotoxicity, demonstrated by reduced ROS levels and lipid peroxidation. Mechanically, B lymphoma Mo-MLV insertion region 1 (Bmi-1) expression and antioxidative superoxide dismutase were increased after RES treatment. Further mechanistic analysis indicated that protection effects of RES against OA-induced lipotoxicity were abrogated by Bmi-1 small interference RNA (siRNA) in L02 cells. SIGNIFICANCE STATEMENT: Results from clinical studies about the effect of RES on NAFLD are inconsistent and inconclusive. This study confirms the protective role of RES as an anti-ROS agent and its ability to alleviate DNA damage through a pathway involving p53/p21 signaling. Further mechanistic analysis indicated that protection effects of RES were relative with Bmi-1. This is the first study on the role of Bmi-1 in the pathogenesis of NAFLD and the target of resveratrol against NAFLD.

BACKGROUND AND AIM:Over 10% of hepatocellular carcinoma (HCC) cases recur each year, even after surgical resection. Currently, there is a lack of knowledge about the causes of recurrence and the effective prevention. Prediction of HCC recurrence requires diagnostic markers endowed with high sensitivity and specificity. This study aims to identify new key proteins for HCC recurrence and to build machine learning algorithms for predicting HCC recurrence. METHODS:The proteomics data for analysis in this study were obtained from the Clinical Proteomics Tumor Analysis Consortium (CPTAC) database. We analyzed different proteins based on cases with or without recurrence of HCC. Survival analysis, Cox regression analysis, and area under the ROC curves (AUROC > 0.7) were used to screen for more significant differential proteins. Predictive models for HCC recurrence were developed using four machine learning algorithms. RESULTS:A total of 690 differentially expressed proteins between 50 relapsed and 77 non-relapsed hepatitis B-related HCC patients were identified. Seven of these proteins had an AUROC > 0.7 for 5-year survival in HCC, including BAHCC1, ESF1, RAP1GAP, RUFY1, SCAMP3, STK3, and TMEM230. Among the machine learning algorithms, the random forest algorithm showed the highest AUROC values (AUROC: 0.991, 95% CI 0.962-0.999) for identifying HCC recurrence, followed by the support vector machine (AUROC: 0.893, 95% Cl 0.824-0.956), the logistic regression (AUROC: 0.774, 95% Cl 0.672-0.868), and the multi-layer perceptron algorithm (AUROC: 0.571, 95% Cl 0.459-0.682). CONCLUSIONS:Our study identifies seven novel proteins for predicting HCC recurrence and the random forest algorithm as the most suitable predictive model for HCC recurrence.

Background and Aims Non-alcoholic fatty liver disease (NAFLD) is a major chronic liver disease worldwide, and non-alcoholic steatohepatitis (NASH) is one of its pathological subtypes. The pathogenesis of NASH has not yet been fully elucidated. The purpose of this study was to identify the hub genes and pathways involved in NASH using bioinformatics methods. The hub genes were confirmed in human and animal models. Materials and Methods Three Gene Expression Omnibus (GEO) datasets (GSE48452, GSE58979, and GSE151158) of NASH patients and healthy controls were included in the study. We used GEO2R to identify differentially expressed genes (DEGs) between NASH patients and healthy controls. Functional enrichment analyses were then performed to explore the potential functions and pathways of the DEGs. In all DEGs, only two genes were highly expressed in NASH patients throughout the three datasets; these two genes, SPP1 and CXCL9, were further studied. Serum and liver tissues from NASH patients and healthy controls were collected. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured in NASH patients and healthy controls. Liver tissues were stained with hematoxylin and eosin. Immunohistochemical staining was used to evaluate the expression levels of the two genes in liver tissues. Male C57BL/6J mice were fed a methionine choline-deficient (MCD) diet for 8 weeks, after which serum ALT and AST levels were measured and liver tissues were stained. Results SPP1 and CXCL9 were the hub genes detected in the three datasets. “Lipid metabolism,” “inflammatory response,” and “lymphocyte activation” were the most significant biological functions in GSE48452, GSE58979, and GSE151158, respectively. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the toll-like receptor signaling pathway was significantly enriched in NASH patients. Serum ALT and AST levels were significantly increased in NASH patients compared to healthy controls. Liver tissues had more serious steatosis, hepatocyte ballooning degeneration, and lobular inflammatory infiltration, and the expression of SPP1 and CXCL9 in liver cells was significantly upregulated in NASH patients compared to healthy controls. MCD diet mice were consistent with NASH patients. Conclusion SPP1 and CXCL9 may play important roles in NASH pathogenesis and could be potential therapeutic targets and biomarkers of NASH in the future. Further experimental studies are needed to confirm our results.

Vitamin D deficiency is common in chronic liver diseases, it may increase the severity of the diseases. However, studies on the potential role of vitamin D in hepatitis B virus (HBV) related end stage liver disease are still limited. This study was performed to assess the prevalence of vitamin D deficiency in patients with different stages of chronic HBV infection and explore whether vitamin D deficiency can be established as an index of severity and prognosticator of disease progression. Serum 25(OH)D3 levels were measured in a cohort of 363 patients with chronic HBV infection. Biochemical parameters, the alpha fetoprotein level, HBV DNA load, hepatitis B surface antigen level, and hepatitis B early antigen level were tested. The mean 25(OH)D3 level was significantly lower in patients with chronic HBV-related liver disease than in healthy controls. Overall, 74.9% of patients had 25(OH)D3 deficiency. The 25(OH)D3 level was significantly positively correlated with the albumin level. In total, 77.6% (121/156) of patients with cirrhosis had vitamin D deficiency. The 25(OH)D3 level significantly decreased as the Child-Pugh classification increased in severity. The 25(OH)D3 level was negatively correlated with the model for end-stage liver disease (MELD) score and low 25(OH)D3 concentrations were significantly associated with high MELD score and mortality in patients with liver failure. Vitamin D deficiency is prevalent in patients with chronic HBV infection. Severe vitamin D deficiency is strongly related to liver dysfunction and disease severity in the cirrhosis and liver failure patients.

To screen out potential prognostic hub genes for adult patients with sepsis via RNA sequencing and construction of a microRNA–mRNA–PPI network and investigate the localization of these hub genes in peripheral blood monocytes. The peripheral blood of 33 subjects was subjected to microRNA and mRNA sequencing using high-throughput sequencing, and differentially expressed genes (DEGs) and differentially expressed microRNAs (DEMs) were identified by bioinformatics. Single-cell transcriptome sequencing (10 × Genomics) was further conducted. Among the samples from 23 adult septic patients and 10 healthy individuals, 20,391 genes and 1633 microRNAs were detected by RNA sequencing. In total, 1114 preliminary DEGs and 76 DEMs were obtained using DESeq2, and 454 DEGs were ultimately distinguished. A microRNA–mRNA–PPI network was constructed based on the DEGs and the top 20 DEMs, which included 10 upregulated and 10 downregulated microRNAs. Furthermore, the hub genes TLR5, FCGR1A, ELANE, GNLY, IL2RB and TGFBR3, which may be associated with the prognosis of sepsis, and their negatively correlated microRNAs, were analysed. The genes TLR5, FCGR1A and ELANE were mainly expressed in macrophages, and the genes GNLY, IL2RB and TGFBR3 were expressed specifically in T cells and natural killer cells. Parallel analysis of mRNAs and microRNAs in patients with sepsis was demonstrated to be feasible using RNA-seq. Potential hub genes and microRNAs that may be related to sepsis prognosis were identified, providing new prospects for sepsis treatment. However, further experiments are needed.

Objective: This study aimed to clarify the infection state of early embryos by HBV and the effect of HBV infection on embryo development. Methods: In vitro culture was carried out on the fresh surplus embryos from infertile couples on the third day (D3) who received routine in vitro fertilization and embryo transfer (IVF-ET) treatment. The embryo cells were infected in vitro by the serum of HBV infected patients with different viral loads. HBsAg quantity in the supernatant secreted by embryo and HBV mRNA in embryo cells were detected. The development of embryo cells after HBV infection was evaluated by embryo morphological parameter score and fragment score in embryo cells. Results: From May 2013 to July 2015, 238 cases of D3 fresh surplus embryos from 120 infertile couples and 128 samples of embryo culture supernatant were collected. The HBsAg positive rate in the supernatant of embryos in the high viral load infection group was significantly higher than that in the low viral load infection group (40.0% vs. 15.6%, p <0.05). HBV mRNA was detected in 2 of 17 HBsAg positive embryos. The scores of embryos in the high load HBV culture solution group were higher than those in the low load group ( p =0.04). There was one formed blastocyst in low load HBV culture group, but there was no blastocyst formation in all embryo cells of high load group ( p <0.001). Conclusion: The results suggest that embryo cells can be directly infected by HBV in vitro. HBV infection affects the development of embryos.