Objective To investigate the role and mechanism of RNA-Specific adenosine deaminase 3 (Adar3) in regulating macrophage polarization during Coxsackievirus B3(CVB3)-induced viral myocarditis (VM). Methods Bone marrow-derived macrophages (BMDM) from mice were cultured in vitro and induced into M1/M2 macrophages using interferon-gamma (IFN-γ)/lipopolysaccharide (LPS) or interleukin 4 (IL-4), respectively. The mRNA expression levels of Adar1, Adar2, and Adar3 in each group of cells were assessed by real-time quantitative PCR (qRT-PCR). Specific siRNAs targeting the Adar3 gene were designed, synthesized, and transiently transfected into M2 macrophages. The mRNA levels of M2 polarization-related marker genes-including arginase 1 (Arg1), chitinase 3-like molecule 3 (YM1/Chi3l3), and resistin-like molecule alpha (RELMα/FIZZ1)-were detected by qRT-PCR. RNA sequencing was performed to analyze the signaling pathways affected by Adar3. The expression levels of Wnt/β-catenin signaling pathway were further validated using qRT-PCR and Western blot. The adeno-associated virus overexpressing Adar3 was designed, synthesized, and injected into mice via tail vein. Three weeks later, a myocarditis mouse model was established. After an additional week, the phenotype and function of cardiac macrophages, as well as multiple indicators of VM (including echocardiography, body weight, histopathology and serology) were examined. Additionally, the protein levels of the Wnt/β-catenin signaling pathway were assessed. Results Compared to M0-type macrophages, the expression level of Adar3 was significantly increased in M2-type macrophages. After transfection of Adar3 siRNA, the mRNA levels of Arg1, YM1 and FIZZ1 in M2 macrophages were downregulated. RNA sequencing revealed 149 upregulated genes and 349 downregulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and subsequent validation experiments indicated that Adar3 modulated the Wnt/β-catenin signaling pathway. In vivo experiments demonstrated that Adar3 overexpression alleviated the cardiac dysfunction of VM mice. The proportion of M1 macrophages in the heart decreased, while the proportion of M2 macrophages increased. At the same time, the Adar3 overexpression activated the Wnt/β-catenin signaling pathway. Conclusion Adar3 promotes macrophage polarization toward the M2 phenotype by activating the Wnt/β-catenin signaling pathway, thereby alleviating VM.
Animals ; Adenosine Deaminase/metabolism* ; Macrophages/immunology* ; Wnt Signaling Pathway/genetics* ; Myocarditis/immunology* ; Mice ; Coxsackievirus Infections/metabolism* ; Male ; Mice, Inbred BALB C ; Enterovirus B, Human/physiology* ; beta Catenin/genetics*

OBJECTIVE: To summarize and analyze the characteristics of delayed hemolytic transfusion reaction in children, in order to provide a scientific basis for clinical prevention, and ensure the safety of children's blood transfusion. METHODS: The basic situation, clinical symptoms and signs, diagnosis time and disappearance time of alloantibody of delayed hemolytic transfusion reaction in children were retrospectively analyzed. The serological test, routine blood test, biochemical detection and urine analysis results were compared pre- and post-transfusion. RESULTS: Among 15 164 children with repeated blood transfusion, 23 cases occurred delayed hemolytic transfusion reactions, with an incidence rate of 0.15%, and mainly children with thalassemia and acute leukemia. 39.13% of delayed hemolytic reactions occurred in children with more than 20 times of blood transfusions. Anemia was the main clinical symptom in 86.96% of children. 4.35% of children had hypotension and dyspnea. Serological test results showed that the positive rate of direct antiglobulin test was 91.30%, and that of erythrocyte homologous antibody test was 100%. Erythrocyte alloantibodies were common in Rh and Kidd blood group systems, accounting for 73.91% and 13.04%, respectively. Laboratory test results showed that hemoglobin, reticulocyte, spherocyte, total bilirubin, indirect bilirubin, lactate dehydrogenase, serum ferritin and urine color were significantly different after transfusion compared with those before transfusion (all P <0.05). The average diagnosis time of delayed hemolytic transfusion reactions was 18.56 days, and the average disappearance time of erythrocyte alloantibodies was 118.43 days. CONCLUSION The incidence of delayed hemolytic transfusion reaction is high in children with repeated blood transfusion, and the disappearance time of erythrocyte homologous antibody is long. Blood matched ABO, Rh and Kidd blood group antigens should be transfused prophylactically. Once diagnosed, erythrocyte alloantibody corresponding to antigen-negative blood should be used throughout the whole process.
Humans ; Child ; Retrospective Studies ; Child, Preschool ; Transfusion Reaction ; Male ; Female ; Infant ; Adolescent ; Isoantibodies/blood* ; Blood Transfusion

Loss-of-function variants in CSDE1 have been strongly linked to neuropsychiatric disorders, yet the precise role of CSDE1 in neurogenesis remains elusive. In this study, we demonstrate that knockout of Csde1 during cortical development in mice results in impaired neural progenitor proliferation, leading to abnormal cortical lamination and embryonic lethality. Transcriptomic analysis revealed that Csde1 upregulates the transcription of genes involved in the cell cycle network. Applying a dual thymidine-labelling approach, we further revealed prolonged cell cycle durations of neuronal progenitors in Csde1-knockout mice, with a notable extension of the G1 phase. Intersection with CLIP-seq data demonstrated that Csde1 binds to the 3' untranslated region (UTR) of mRNA transcripts encoding cell cycle genes. Particularly, we uncovered that Csde1 directly binds to the 3' UTR of mRNA transcripts encoding Cdk6, a pivotal gene in regulating the transition from the G1 to S phases of the cell cycle, thereby maintaining its stability. Collectively, this study elucidates Csde1 as a novel regulator of Cdk6, sheds new light on its critical roles in orchestrating brain development, and underscores how mutations in Csde1 may contribute to the pathogenesis of neuropsychiatric disorders.
Animals ; Neurogenesis/genetics* ; Cell Cycle/genetics* ; Mice, Knockout ; Mice ; Neural Stem Cells/metabolism* ; DNA-Binding Proteins/metabolism* ; Cyclin-Dependent Kinase 6/genetics* ; Cell Proliferation ; 3' Untranslated Regions ; Cerebral Cortex/embryology* ; RNA-Binding Proteins ; Mice, Inbred C57BL

Objective To explore the diagnostic value of mediastinal lymph nodes CT image features in pulmonary sarcoidosis.Methods The imageologic data of the patients with pulmonary sarcoidosis(pulmonary sarcoidosis group)and lung cancer complicating mediastinal lymph node metastasis(lung cancer group)con-firmed by EBUS-TBNA in this hospital from June 2015 to November 2023 were analyzed retrospectively.The chest plain scan CT and enhanced scan CT were performed in all cases,and the imaging characteristics were compared between the two groups.The receiver operating characteristic(ROC)curve was drawn and the area under the curve(AUC)was calculated.The diagnostic value of CT imaging characteristics in pulmonary sar-coidosis was analyzed.Results The proportion of female patients in the lung sarcoidosis group was signifi-cantly higher than that in the lung cancer group,while the age was smaller than that in the lung cancer group,and the differences were statistically significant(P＜0.05).The number of mediastinal lymph nodes,location,short diameter and CT enhancement value in the pulmonary sarcoidosis group had statistical difference(P＜0.05).The ROC curve analysis showed that AUC of lymph node short diameter reciprocal,CT enhancement value and position in detection alone were 0.586,0.785 and 0.505 respectively,and AUC of the three combined reciprocal was the highest(0.789).Conclusion The lymph node short diameter,CT enhancement value and po-sition have certain value in clinical diagnosis of pulmonary sarcoidosis and the 3-indicator combination could increase the diagnostic efficiency of pulmonary sarcoidosis.

Objective To analyze clinical characteristics affecting survival outcomes in gastric adenocarcinoma(GAC)patients with second primary malignancies(SPM)and construct a predictive model with a web-based calculator.Methods Patients diagnosed with GAC between January 2010 and December 2017 in the SEER database(n=24 085)were analyzed,comparing non-SPM(n=22 963)and SPM cohorts(n=1 122).SPM patients were randomized(3:1)into training(n=842)and internal validation cohorts(n=280).Univariate/multivariate Cox regression identified prognostic factors for model construction.Model performance was evaluated via ROC curves,calibration plots,and decision curve analysis(DCA).A web-based calculator was deployed using DynNom(https://kunma697.shinyapps.io/dynnomapp-1/).External validation used 192 SPM patients diagnosed at Yantai Yuhuangding Hospital(2010-2017).Results χ2 tests revealed SPM patients had higher age(56.3%),earlier T-stage(T1:29.2%;T2:10.5%),predominant gastric cardia involvement(43.7%),fewer distant metastases(12.3%),and higher rates of radiotherapy(32.5%)and surgery(77.2%)vs.non-SPM(P<0.05).Cox analyses identified GAC primary site,T-stage,SEER stage,radiotherapy/surgery history,plus SPM grade/stage/treatment history as significant predictors(P<0.05).AUCs in the training cohort were 0.771(95%CI:0.722～0.820),0.839(95%CI:0.796～0.882),and 0.836(95%CI:0.792～0.879)for 1-/3-/5-year survival;internal validation showed 0.751(95%CI:0.700～0.801),0.746(95%CI:0.695～0.797),and 0.772(95%CI:0.723～0.821);external validation yielded 0.713(95%CI:0.648～0.778),0.805(95%CI:0.749～0.861),and 0.851(95%CI:0.801～0.901).Calibration indicated high prediction-actuality concordance;DCA confirmed clinical utility.Conclusion The model and web calculator incorporating GAC/SPM characteristics effectively predict SPM patient prognosis.

Objective:To study the role and preliminary molecular mechanism of TRIM24 regulating macrophage polarization in viral myocarditis(VM).Methods:VM mouse model was established by Coxsackie virus B3(CVB3),and expression of TRIM24 in myocardial tissue was detected.Cardiac inflammation level and polarization phenotype of cardiac infiltrating macrophages in a murine model of cardiac TRIM24 inhibition were detected in vivo.A polarization model of mouse bone marrow-derived macrophages(BMDMs)in vitro was established to observe the role of TRIM24 inhibition in polarizing BMDMs to M1 and M2,as well as its effects on phagocy-tosis and bactericidal function of BMDMs.Effects of TRIM24 inhibition on total STAT6 protein level and phosphorylation were investi-gated.Results:TRIM24 was significantly highly expressed in myocardial tissue of VM mice(P<0.001).Inhibition of TRIM24 expres-sion in myocardium had an attenuating effect on VM and promoted polarization of cardiac infiltrating macrophages to M2.TRIM24 was significantly down-regulated in vitro during the polarization of BMDMs toward M2(P<0.01).Inhibition of TRIM24 expression signifi-cantly promoted macrophage polarization toward M2 type and inhibited polarization toward M1 type,accompanied by a significant increase in STAT6 phosphorylation levels(P<0.01).Conclusion:TRIM24 regulates macrophage M2 polarization via activation of STAT6 signaling pathway to attenuate VM.

Background Atmospheric fine particulate matter (PM_2.5) can disrupt the metabolic homeostasis of the liver and accelerate the progression of liver diseases, but there are few studies on the effects of sub-chronic PM_2.5 exposure on the liver metabolome. Objectives To investigate the effects of sub-chronic exposure to concentrated PM_2.5 on hepatic metabolomics in mice by liquid chromatography-mass spectrometry (LC-MS), and to identify potentially affected metabolites and metabolic pathways. Methods Twelve male C57BL/6J (6 weeks old) mice were randomly divided into two groups: a concentrated PM_2.5 exposure group and a clean air exposure group. The mice were exposed to concentrated PM_2.5 using the "Shanghai Meteorological and Environmental Animal Exposure System" at Fudan University. The exposure duration was 8 h per day, 6 d per week, for a total of 8 weeks. The mice's liver tissues were collected 24 h after the completion of exposure. LC-MS was performed to assess changes in the hepatic metabolome. Orthogonal partial least squares discriminant analysis and t-test were employed to identify differentially regulated metabolites between the two groups under the conditions of variable important in projection (VIP)≥1.0 and P<0.05. Metabolic pathway enrichment analysis was performed using MetaboAnalyst 5.0 software and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Results A total of 297 differentially regulated metabolites were identified between the concentrated PM_2.5 exposure group and the clean air group. Among these metabolites, 142 were upregulated and 155 were downregulated. A total of 38 metabolic pathways were altered, with 7 pathways showing significant perturbation (P<0.05). These pathways involved amino acid metabolism, glucose metabolism, nucleotide metabolism, as well as cofactor and vitamin metabolism. The 7 significant metabolic pathways were pantothenic acid and coenzyme A biosynthesis; purine metabolism; amino sugar and nucleotide sugar metabolism; arginine biosynthesis; alanine, aspartate and glutamate metabolism; aminoacyl-tRNA biosynthesis; and fructose and mannose metabolism. Conclusion The results from metabolomics analysis suggest that sub-chronic exposure to PM_2.5 may disrupt hepatic energy metabolism and induce oxidative stress damage. Aspartic acid, succinic acid, ornithine, fumaric acid, as well as purine and xanthine derivatives, were identified as potential early biomarkers of hepatic response to sub-chronic PM_2.5 exposure.

Objective:To explore the diagnosis and treatment of lymphoepithelioma-like intrahepatic cholangiocarcinoma(LEL-ICC).Methods:The retrospective and descriptive study was conducted. The data of 7 patients with pathological diagnosis of LEL-ICC after hepatectomy who were treated in Lishui Central Hospital in Zhejiang Province from December 1, 2009 to January 30, 2024 were collected. There were 2 males and 5 females. The age range was from 40 to 64 years old, with a median age of 52 years old. All 7 patients showed no obvious clinical symptoms.We analysed the imaging manifestations, pathological features, treatmentsand prognoses of patients.Postoperative follow-upswere conducted via telephone, with a focus on whether the patient had relapsed. The deadline was February 20, 2024.Results:Five cases underwent ultrasound examination, of which 4 cases showed hypoechogenicity and 1 case showed hyperechogenicity. 7 cases underwent MRI examination, showing low signal on T1WI, high signal on T2WI, and high signal on diffusion-weighted imaging. 2 cases had type A enhancement, 2 cases had type B enhancement, and 3 cases had type C enhancement. All 7 cases received surgical treatment, 2 cases were received prophylactic transarterial chemoembolization (TACE) after surgery, and 3 cases were received systemic chemotherapy after surgery; All 7 cases underwent postoperative follow-up, with a follow-up time of 1-166 months and a median follow-up time of 56 months. One case developed hilar and retroperitoneal lymph node metastasis after surgery for 6 months, and underwent surgical treatment. After surgery, chemotherapy was performed. 25 months later, right adrenal gland metastasis reappeared, and after combined treatment, the metastatic lesion was reduced and the patient received surgical treatment and chemotherapy, and there is currently no recurrence. The remaining 6 cases showed no recurrence.Conclusions:LEL-ICC lacks specific clinical symptoms and imaging manifestations, diagnosis relies on histopathological and immunohistochemical examinations. Comprehensive treatment with surgical intervention as the main approach can lead to better prognosis for patients.

Objective:To explore the clinical manifestations, diagnosis and treatment methods, and prognosis of hepatic portal venous gas (HPVG).Methods:Retrospective case analysis was used in the case data of 7 patients with HPVG, who were treated in Lishui Central Hospital from January 2017 to July 2024, including 5 males and 2 females. Age ranged from 46 to 90 years, with an average age of 69 years. Abdominal pain was the first manifestation in 6 cases, and septic shock occurred in 2 cases. The initial symptoms, primary diseases, comorbidities, laboratory results, imaging examinations, treatment plans, and prognosis were analyzed. Prognostic follow-up was conducted by telephone, with the focus on whether the patient had experienced HPVG recurrence and postoperative complications. The deadline for follow-up was July 31, 2024.Results:All patients had elevated white blood cells and C-reactive protein (CRP), and prothrombin time was prolonged in 4 patients. pH and base excess decreased in 4 cases, and lactic acid increased in 5 cases. Alanine aminotransferase increased in 2 cases, and total bilirubin increased in 3 cases. Blood culture was positive in 3 cases. Contrast-enhanced abdominal CT showed clear gas shadows in the portal vein and its branches in all 7 cases, which were confined to the left liver in 4 cases and distributed in both sides of the liver in 3 cases. The primary diseases were intestinal obstruction and necrosis in 4 cases, intestinal perforation and necrosis in 1 case, inflammatory bowel disease in 1 case, and acute pancreatitis in 1 case. Five patients were complicated with hypertension, diabetes and other diseases. Two patients received surgical treatment, and the average time from surgery to diagnosis was 6.8 h. Five cases received conservative treatment, 2 cases were cured and 3 cases died, with an average time from onset to death of 21.2 h.Conclusions:Contrast-enhanced abdominal CT should be the preferred method for the diagnosis of HPVG. The primary disease and its severity should be fully evaluated in the treatment of HPVG. Patients with intestinal necrosis should undergo laparotomy as soon as possible, and enterostomy should be performed during the operation. Clinicians should strengthen the awareness of HPVG in order to improve the prognosis of patients.

Objective To compare the outcomes of transplant kidneys and patient survival between simultaneous pancreas-kidney transplantation(SPKT)recipients and deceased donor kidney transplant(DDKT)recipients in patients with type 2 diabetes mellitus(T2DM)complicated with end-stage renal disease(ESRD),and to analyze the risk factors affecting patient survival post-SPKT.Methods Clinical and prognostic data of patients who underwent kidney transplantation from January 27,2003,to January 1,2021,were retrieved from the United Network for Organ Sharing(UNOS)database.A total of 50 230 cases were selected based on inclusion criteria,with 48 669 cases in DDKT group and 1561 cases in SPKT group.Kaplan-Meier analysis was employed to compare transplant kidney and patient survival between the two groups,and propensity score matching(PSM)was utilized to balance confounding factors between the groups.Cox regression model was used to analyze independent risk factors affecting patient survival post-SPKT.Results Compared with DDKT group,recipients in SPKT group had a younger median age(P<0.001),a higher proportion of males(P<0.001),lower BMI(P<0.001),shorter dialysis and transplant waiting times(P<0.001),a higher percentage of private medical insurance(P<0.001),a lower proportion of previous transplants(P<0.001),a younger age at diabetes diagnosis(P<0.001),and a lower incidence of peripheral vascular disease(P=0.033).Compared with DDKT group,the donors in SPKT group had a younger median age(P<0.001),a higher proportion of males(P<0.001),lower BMI(P<0.001),and a lower prevalence of hypertension and diabetes history(P<0.001).In terms of transplant-related factors,the SPKT group had a shorter donor kidney cold ischemia time(P<0.001),a higher degree of HLA mismatch(P<0.001),and a lower Kidney Donor Profile Index(KDPI)(P<0.001)when compared with DDKT group.The SPKT group had lower serum creatinine levels at discharge(P<0.001),lower rates of postoperative delayed graft function(DGF)and acute rejection(AR)(P<0.001),but longer hospital stays(P<0.001)when compared with DDKT group.Kaplan-Meier survival analysis curves,both original and after propensity score matching(PSM),consistently showed significantly higher transplant kidney and patient survival rates in SPKT group compared with DDKT group(P<0.001).Cox regression model analysis indicated that recipient age,recipient race,donor age,and donor kidney cold ischemia time were independent risk factors influencing patient survival post-SPKT.Conclusions For ESRD patients with T2DM,SPKT offers improved long-term graft and patient survival rates compared with DDKT.Recipient age,recipient ethnicity,donor age,and cold ischemia time for the donor's kidney are independent risk factors affecting post-SPKT patient survival.