Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive hematologic malignancy with a poor prognosis, despite advancements in treatment. Many patients struggle with relapse or refractory disease. Investigating the role of the super-enhancer (SE) regulated gene ubiquitin-specific protease 20 (USP20) in T-ALL could enhance targeted therapies and improve clinical outcomes. Analysis of histone H3 lysine 27 acetylation (H3K27ac) chromatin immunoprecipitation sequencing (ChIP-seq) data from six T-ALL cell lines and seven pediatric samples identified USP20 as an SE-regulated driver gene. Utilizing the Cancer Cell Line Encyclopedia (CCLE) and BloodSpot databases, it was found that USP20 is specifically highly expressed in T-ALL. Knocking down USP20 with short hairpin RNA (shRNA) increased apoptosis and inhibited proliferation in T-ALL cells. In vivo studies showed that USP20 knockdown reduced tumor growth and improved survival. The USP20 inhibitor GSK2643943A demonstrated similar anti-tumor effects. Mass spectrometry, RNA-Seq, and immunoprecipitation revealed that USP20 interacted with hypoxia-inducible factor 1 subunit alpha (HIF1A) and stabilized it by deubiquitination. Cleavage under targets and tagmentation (CUT&Tag) results indicated that USP20 co-localized with HIF1A, jointly modulating target genes in T-ALL. This study identifies USP20 as a therapeutic target in T-ALL and suggests GSK2643943A as a potential treatment strategy.

OBJECTIVE: To develop a method for identifying high-risk patients among septic populations requiring mechanical ventilation, and to conduct phenotypic analysis based on this method. METHODS: Data from four sources were utilized: the Medical Information Mart for Intensive Care (MIMIC-IV 2.0, MIMIC-III 1.4), the Philips eICU-Collaborative Research Database 2.0 (eICU-CRD 2.0), and the Anhui Medical University Second Affiliated Hospital dataset. The adult patients in intensive care unit (ICU) who met Sepsis-3 and received invasive mechanical ventilation (IMV) on the first day of first admission were enrolled. The MIMIC-IV dataset with the highest data integrity was divided into a training set and a test set at a 6:1 ratio, while the remaining datasets were served as validation sets. The demographic information, comorbidities, laboratory indicators, commonly used ICU scores, and treatment measures of patients were extracted. Clinical data collected within first day of ICU admission were used to calculate the sequential organ failure assessment (SOFA) score. K-means clustering was applied to cluster SOFA score components, and the sum of squared errors (SSE) and Davies-Bouldin index (DBI) were used to determine the optimal number of disease subtypes. For clustering results, normalized methods were employed to compare baseline characteristics by visualization, and Kaplan-Meier curves were used to analyze clinical outcomes across phenotypes. RESULTS: This study enrolled patients from MIMIC-IV dataset (n = 11 166), MIMIC-III dataset (n = 4 821), eICU-CRD dataset (n = 6 624), and a local dataset (n = 110), with the four datasets showing similar median ages and male proportions exceeding 50%; using 85% of the MIMIC-IV dataset as the training set, 15% as the test set, and the rest dataset as the validation set. K-means clustering based on the six-item SOFA score was performed to determine the optimal number of clusters as 3, and patients were finally classified into three phenotypes. In the training set, compared with the patients with phenotype II and phenotype III, those with phenotype I had the more severe circulatory and respiratory dysfunction, a higher proportion of vasoactive drug usage, more obvious metabolic acidosis and hypoxia, and a higher incidence of congestive heart failure. The patients with phenotype II was dominated by respiratory dysfunction with higher visceral injury. The patients with phenotype III had relatively stable organ function. The above characteristics were consistent in both the test and validation sets. Analysis of infection-related indicators showed that the patients with phenotype I had the highest SOFA score within 7 days after ICU admission, initial decreases and later increases in platelet count (PLT), and higher counts of neutrophils, lymphocytes, and monocytes as compared with those with phenotype II and phenotype III, their blood cultures had a higher positivity rates for Gram-positive bacteria, Gram-negative bacteria and fungi as compared with those with phenotype II and phenotype III. The Kaplan-Meier curve indicated that in the training, test, and validation sets, the 28-day cumulative mortality of patients with phenotype I was significantly higher than that of patients with phenotypes II and phenotype III. CONCLUSIONS Three distinct phenotypes in septic patients receiving IMV based on unsupervised machine learning is derived, among which phenotype I, characterized by cardiorespiratory failure, can be used for the early identification of high-risk patients in this population. Moreover, this population is more prone to bloodstream infections, posing a high risk and having a poor prognosis.
Humans ; Machine Learning ; Sepsis/therapy* ; Respiration, Artificial ; Intensive Care Units ; Organ Dysfunction Scores ; Male ; Female ; Middle Aged ; Adult

Objective:To investigate the relationship between preoperative plasma fibrin degradation products (FDP) level and clinicopathological features of patients with completely resected non-small cell lung cancer (NSCLC).Methods:A retrospective case series study was performed. The clinical data of 521 patients who were pathologically diagnosed with NSCLC in Beijing Friendship Hospital Affiliated to Capital Medical University from January 2016 to December 2017 were retrospectively analyzed. Among 521 cases, 406 cases were postoperatively pathologically confirmed as non-lymph node and non-distant metastasis (non-metastasis group) and 115 cases were postoperatively pathologically confirmed as lymph node or distant metastasis (metastasis group). The preoperative FDP level and clinicopathological characteristics as well as the clinicopathological characteristics of NSCLC patients with different FDP levels were compared between the two groups. The correlation between preoperative FDP level and TNM staging was analyzed by using Spearman correlation analysis.Results:Among 521 NSCLC patients, 266 cases were female, 255 cases were male; the age [ M( Q1, Q3)] was 59 years (54 years, 65 years); 441 cases were adenocarcinoma and 70 cases were squamous cell carcinoma. The preoperative median FDP level was 2.78 mg/L (2.35 mg/L, 3.13 mg/L) and 2.99 mg/L (2.56 mg/L, 4.16 mg/L), respectively of NSCLC patients in non-metastasis group and metastasis group, and the difference was statistically significant ( Z = 6.13, P < 0.001). The preoperative FDP level was 2.56 mg/L (2.35 mg/L, 3.20 mg/L) and 2.99 mg/L (2.56 mg/L, 3.20 mg/L), respectively in the early-stage NSCLC (stage Ⅰ-Ⅱ) and advanced NSCLC (stage Ⅲ-Ⅳ) patients, and the difference was statistically significant ( Z = 8.42, P < 0.001). Spearman correlation analysis showed that preoperative FDP level was positively correlated with tumor diameter ( r = 0.287, P < 0.001). There was a positive correlation between preoperative FDP level and the number of metastatic lymph nodes in 115 patients with lymph node metastasis ( r = 0.679, P < 0.001). According to the preoperative median FDP (2.78 mg/L), all patients were divided into FDP ≤2.78 mg/L group and FDP >2.78 mg/L, and there were statistically significant differences in age, metastasis, tumor staging, tumor diameter, the metastatic number of lymph node and histological types of NSCLC patients in both groups (all P < 0.05). Conclusions:The increase of preoperative plasma FDP level may be related to the tumor metastasis and clinical stage of NSCLC patients

Objective:To investigate the current status of malnutrition and its influential factors among patients with stroke and dysphagia, and to develop and validate a malnutrition risk prediction model.Methods:Using a convenience sampling method, 150 patients with stroke and dysphagia admitted to Wenzhou Central Hospital from January 2019 to December 2023 were included in this study. Through a review of the literature and expert consultations, 15 influential factors were identified: age, gender, body mass index (BMI), history of smoking alcohol consumption , number of hospitalizations, education level, Barthel index, history of hypertension, history of diabetes, coronary heart disease, presence of limb disabilities, hemoglobin levels, Glasgow Coma Scale (GCS) score, and National Institutes of Health Stroke Scale score. Patients were categorized into malnutrition and normal groups based on the occurrence of malnutrition. The influential factors for malnutrition were analyzed, and a malnutrition risk prediction model was constructed using regression analysis. The model was presented using a nomogram and subsequently validated.Results:Among the 150 patients with stroke and dysphagia, the average age was (59.34 ± 6.46) years, with 83 females and 67 males. Of these patients, 66 (44.00%) were found to be malnourished. The following factors were identified as independent risk factors for malnutrition in patients with stroke and dysphagia: age (χ2 = 4.03, P = 0.045), BMI ( t = 6.33, P < 0.001), alcohol consumption (χ2 = 3.90, P = 0.048), number of hospitalizations (χ2 = 9.45, P = 0.024), Barthel index (χ2 = 7.78, P = 0.020), presence of limb disabilities (χ2 = 4.64, P = 0.031), hemoglobin levels (χ2 = 4.38, P = 0.036), and GCS score (χ2 = 9.83, P = 0.007) (all P < 0.05). Patients who were older, had a BMI < 18.5 kg/m2, consumed alcohol, had more than five hospitalizations, a Barthel index < 40, limb disabilities, abnormal hemoglobin levels, or a GCS score ≤ 11 were more likely to experience malnutrition (all P < 0.05). The C-index for predicting malnutrition was 0.851, with a 95% CI of (0.809, 0.892). The maximum Youden index was 0.562, with a sensitivity of 84.1% and specificity of 72.1%. Conclusion:The risk factors for malnutrition in patients with stroke and dysphagia include advanced age, alcohol consumption, more than five hospitalizations, limb disabilities, and abnormal hemoglobin levels. Protective factors against malnutrition in these patients are a BMI > 23.9 kg/m2, a Barthel index > 60, and a GCS score ≥ 14. The prediction model demonstrates a significant predictive value for the occurrence of malnutrition in patients with stroke and dysphagia.

Cases of high-temperature requirement A serine peptidase 1(HTRA1)gene heterozygous mutation associated cerebral small vessel disease are relatively rare.Early and timely diagnosis and treatment can improve prognosis.The authors reported a 37 years old male patient admitted in Department of Neurology,the Second Affiliated Hospital of Nanchang University,whose initial symptom was transient right limb weakness,imaging suggested white matter lesions and the gene screening showed HTRA1(c.854 C＞T/p.Pro285Leu)heterozygous mutation.A family survey has been conducted and the characteristics of patients in this family are as follows:they present with ischemic cerebrovascular disease,coexist with cervical or lumbar disc herniation,male patients have hair loss,some patients have cognitive dysfunction,men tend to develop the disease at an earlier age than women,and the onset age is progressively earlier from generation to generation.Therefore,for young ischemic cerebrovascular disease patients with hair loss,cognitive dysfunction,cervical or lumbar disc herniation,and obvious white matter lesions on imaging,especially those without common risk factors for cerebrovascular disease,a family history should be inquired and genetic testing should be performed to screen for HTRA1 mutations.

Objective We investigated the effects of ICAM5 in the hippocampus on the alcohol drinking preference of mice,and the potential mechanisms.Methods An alcohol two-bottle choice model was developed in 8-week-old male C57BL/6J mice,which were randomly divided to two groups:water group and alcohol group.The protein expression of ICAM5 in the hippocampus,amygdala,and medial prefrontal cortex was detected.An ICAM5-overexpressing adeno-associated virus was constructed and injected into the hippocampus by stereotaxic method.The expression level of ICAM5 protein in the hippocampus was detected by immunofluorescence and Western blot.We then detected the alcohol preference and locomotor activity of mice with a conditioned place preference(CPP)experiment,open field test,and loss-of-righting reflex test.Western blot analysis was used to identify the neuron F-actin/G-actin ratio.Using Golgi staining,the morphology of dendritic spines was identified.Results The expression of ICAM5 in the hippocampus of alcohol two-bottle choice model mice in the alcohol group was considerably lower than that of the water group(P＜0.001).The specific expression of ICAM5 in the hippocampus of mice was observed by fluorescence microscopy.In the open field experiment,the staying time and moving distance of the AAV-ICAM5 group were significantly increased compared with those of the control group(P＜0.01).In the CPP experiment,the residence time of AAV-ICAM5 mice in the alcohol-paired compartment was significantly lower than that of control mice(P＜0.001).In the loss-of-righting reflex experiment,overexpression of ICAM5 significantly reduced sedation latency(P＜0.01),but significantly shortened the duration of sedation(P＜0.001).Compared with AAV-mCherry+Water group,the ratio of F-actin/G-actin in the hippocampus was significantly increased after drinking(P＜0.01),but after ICAM5 overexpression,their F-actin/G-actin ratio was significantly decreased(P＜0.001).Compared with AAV-mCherry+Water group,the density of dendritic spines in the hippocampal CAI region was increased(P＜0.001),but the density of dendritic spines in the AAV-ICAM5+Alcohol group was significantly decreased(P＜0.01).Conclusions ICAM5 modulated the expression of cytoskeleton proteins to change the structural plasticity of dendritic spines,which contributed to alcohol-drinking and locomotor behavioral changes in mice.

AIM:To investigate the impact of hippocampal protein phosphatase 1 regulator subunit 17(Ppp1r17)down-regulation on drinking-related behavior in mice,and to explore the protein kinase B(PKB/AKT)/glyco-gen synthase kinase-3β(GSK-3β)/cAMP response element binding protein(CREB)signaling pathway of Ppp1r17 in the development of alcohol dependence.METHODS:Forty male C57BL/6J mice were randomly divided into four groups(n=10):control group,shPpp1r17① group,shPpp1r17② group and shPpp1r17③ group.The mRNA and protein expression levels in the hippocampal tissues were evaluated after 3 weeks of AAV-shPpp1r17 injection.Twenty male C57BL/6J mice were randomly divided into a control group and a shPpp1r17 group(n=10).An open-field test,conditioned place prefer-ence(CPP)test and righting reflex test were performed.Furthermore,the localization and expression of AAV-shPpp1r17 were detected after 3 weeks of AAV-shPpp1r17 injection.Twenty male C57BL/6J mice were randomly divided into four groups(n=5):the empty virus+water group,the empty virus+EtOH group,the shPpp1r17+water group and the shPpp1r17+EtOH group.The EtOH group mice drank 9%alcohol continuously for 30 d.The protein expression levels of Ppp1r17,p-AKT,AKT,p-GSK-3β,GSK-3β,p-CREB and CREB were detected by Western blot.RESULTS:(1)The results of qPCR showed that Ppp1r17 was successfully suppressed by shPpp1r17.(2)The behavioral results showed that the shPpp1r17 group mice exhibited enhanced exercise ability and reduced anxiety-like emotions,and the downregulation of Ppp1r17 increased CPP scores and reduced the sensitivity of the mice to alcohol.(3)Immunofluorescence showed that AAV-shPpp1r17 was specifically expressed in the hippocampus.(4)Western blot analysis revealed that Ppp1r17 and the p-AKT/AKT,p-GSK-3β/GSK-3β and p-CREB/CREB ratios were significantly increased after treatment of EtOH.Howev-er,the protein expression of Ppp1r17 was significantly reduced,and the AKT/GSK-3β/CREB pathway was activated after knockdown of Ppp1r17 in the hippocampus.CONCLUSION:Ppp1r17 may suppress CREB phosphorylation through the AKT/GSK-3β/CREB pathway,thereby influencing alcohol drinking preference and locomotor behavior in mice.