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 investigate the relationship between the cerebral small vascular disease (CSVD) total burden and the imaging markers and the degree of unilateral middle cerebral artery (MCA) stenosis.Methods:The study was a cross-sectional study. Clinical and imaging data of patients with chronic unilateral MCA stenosis who underwent multimodal MRI from October 2015 to January 2019 in the First Medical Center of PLA General Hospital were retrospectively analyzed. A total of 261 patients were included, 187 males and 74 females. According to the degree of MCA stenosis, the patients were divided into 102 cases in severe stenosis-occlusion group (stenosis degree ≥70%) and 159 cases in mild-moderate stenosis group (stenosis degree <70%). CSVD imaging marker scores (including white matter hyperintensity, perivascular space, cerebral microbleed, and lacune of presumed vascular origin) were assessed according to the ?standards for reporting vascular changes on neuroimaging 1 in the 2 groups, and the CSVD total burden score was calculated. Mann-Whitney U test was used to compare the indicators between the two groups, and the CSVD total burden score and imaging marker scores were ultimately included in a multifactorial binary logistic regression to assess the association of CSVD imaging markers with severe stenosis-occlusion of the MCA after adjusting for vascular risk factors (age, gender, drinking, smoking, hypertension, hyperlipidemia, atrial fibrillation and coronary heart disease). Results:There were significant differences in the CSVD total burden, centrum semiovale perivascular space and lacune of presumed vascular origin score between the mild-to-moderate stenosis group and the severe stenosis-occlusion group (all P<0.05), and none of the differences in the remaining imaging marker scores were statistically significant (all P>0.05). Multivariate binary logistics regression analysis showed CSVD total burden score ( OR=1.300, 95% CI 1.047-1.613, P=0.017), centrum semiovale perivascular space score ( OR=2.099, 95% CI 1.540-2.860, P<0.001) and lacune of presumed vascular origin score ( OR=2.609, 95% CI 1.294-5.261, P=0.007) were independent associated with severe stenosis-occlusion of MCA. Conclusion:The higher CSVD total burden score, centrum semiovale perivascular space score and lacune of presumed vascular origin score are associated with severe stenosis-occlusion of MCA.

Objective To investigate the therapeutic effects and mechanisms of Maxing Shigan Decoction on cough variant asthma(CVA)rats.Methods Sixty rats were randomly divided into normal group,model group,low and high dose groups of Maxing Shigan Decoction,and high-dose of Maxing Shigan Decoction + signal transducer and activator of transcription 3(STAT3)activator Colivelin(Col)group,12 rats in each group.Except for the normal group,the CVA model was constructed by intraperitoneal injection of ovalbumin combined with moxa fumigation in all other groups of rats.After the corresponding treatment,the rats were observed for signs and cough counts,airway resistance(RE)was detected by pulmonary function meter,eosinophils(EOS)were counted by Diff-Quik staining,histopathological features of the lungs and bronchial tubes were observed by hematoxylin-eosin(HE)staining method,and the lung tissues were detected by enzyme-linked immunosorbent assay(ELISA)for monocyte chemotactic protein 1(MCP-1),and tumor necrosis factor α(TNF-α),and the protein expression levels of interleukin 6(IL-6),STAT3,and transient receptor potential vanilloid-1 channel(TRPV1)were detected by Western Blot.Results Compared with the normal group,rats in the model group showed obvious asthma symptoms,severe inflammatory cell infiltration was seen in the lung tissue,bronchial epithelial cell necrosis,ciliated adhesion,mucus,and RE,EOS number,MCP-1 and TNF-α contents,and protein expression levels of IL-6,STAT3,TRPV1 were elevated(P＜0.05);compared with the model group,rats in the low-and high-dose groups of Maxing Shigan Decoction showed significant improvement in asthma symptoms,reduction in lung and bronchial injury,and dose-dependent reduction in RE,EOS number,MCP-1 and TNF-α contents,and protein expression levels of IL-6,STAT3,and TRPV1(P＜0.05);compared with the high-dose group of Maxing Shigan Decoction,the rats in the high-dose Maxing Shigan Decoction+Col group showed increased asthma,increased lung and bronchial injury,and increased RE,EOS number,MCP-1 and TNF-α contents,and protein expression levels of IL-6,STAT3,and TRPV1(P＜0.05).Conclusion Maxing Shigan Decoction can effectively improve cough variant asthma in rats,and its mechanism is related to the inhibition of IL-6/STAT3 signaling pathway and the high expression of TRPV1.

Objective To investigate the possible role and mechanism of activation of pyroptosis classical pathway and alterations in cell adhesion in calcium-containing kidney stones after the action of high concentration of Ca2+ on HK-2 cells.Methods HK-2 cells were cultured in the presence of different concentrations of CaCl2(0,0.1,0.5,1.0,2.0,4.0 and 8.0 g/L)for 24 hours,and cell counting Kit-8(CCK-8)and flow cytometry were used to determine the optimal treatment concentration.Subsequently,the ultrastructure of renal tubular epithelial cells under high Ca2+ condition was observed by transmission electron microscopy after Ca2+ treatment.DCFH-DA staining was used to detect intracellular reactive oxygen species production,and quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot analysis were performed to examine the expression of pyroptosis-related proteins NLRP3,Caspase-1,gasdermin D(GSDMD),adhesive molecules osteopontin(OPN)and CD44 at mRNA and protein levels after high concentration Ca2+ treatment.The expression levels of pyroptosis-related inflammatory factors interleukin(IL)-1β,IL-18 and adhesive molecule monocyte chemotactic protein-1(MCP-1)were detected by enzyme-linked immunosorbent assay(ELISA)after high Ca2+ stimulation.Results Ca2+ showed cytotoxicity for HK-2 cell growth and can promote apoptosis.The higher the Ca2+ concentration,the more toxicity and apoptosis rate for HK-2 cell growth.High concentration of Ca2+ can promote pyroptosis-like morphological changes in HK-2 cells,including loss of cell membrane integrity,release of contents and numerous intracellular vacuoles.Compared with the control group,the expression levels of ROS were sequentially increased in the 1.0 g/L CaCl2 group and the 2.0 g/L CaCl2 group,and the expression levels of pyroptosis-related genes NLRP3,Caspase-1,GSDMD,and the pyroptosis-associated inflammatory factors IL-1β and IL-18,as well as the adhesion molecules OPN,CD44 and MCP-1 were significantly increased(P＜0.05).Conclusion High Ca2+ treatment can cause oxidative stress damage in HK-2 cells to produce ROS,which activates NLRP3 inflammasome,leads to the activation of the classical pathway of pyroptosis and increase the adhesion of cells,and ultimately leads to the formation of kidney stones.

BACKGROUND:A large number of domestic and international documents have confirmed that elevated interleukin-1β is associated with primary frozen shoulder.Interleukin-1B gene polymorphisms can affect the transcription and protein expression of interleukin 1β-related genes,resulting in altered levels of cytokines in vivo,and thus altering the incidence of primary frozen shoulder.Through the study of interleukin-1B gene polymorphism and susceptibility to primary frozen shoulder,this study aimed to explore new breakthroughs in the pathogenesis of primary frozen shoulder from the perspective of molecular biology,and to search for susceptibility genes of primary frozen shoulder. OBJECTIVE:To explore the association between linkage disequilibrium of three gene loci in interleukin-1B gene and susceptibility to primary frozen shoulder. METHODS:A case-control study was conducted.There were two groups in this study.One group consisted of 184 patients with primary frozen shoulder,while the other group included 260 healthy controls.The genotypes of interleukin-1B gene loci-511C/T(rs16944),+3954C/T(rs1143634),and-31C/T(rs1143627)were detected by polymerase chain reaction and restriction fragment length polymorphism.The correlation between the probability of linkage disequilibrium and haplotypes and the risk of primary frozen shoulder disease was compared and analyzed. RESULTS AND CONCLUSION:Unconditional Logistic regression analysis showed that the proportion of CT genotypes at rs1143634 and rs1143627 sites increased significantly in the primary frozen shoulder.Linkage disequilibrium analysis showed that rs16944,rs1143634 and rs1143627 tended to be balanced in the control group(D'value<0.1),while there was a certain degree of linkage disequilibrium at rs1143627 and rs1143634 sites in the primary frozen shoulder group(D'value=0.595).Haplotype TTT increased the risk of primary frozen shoulder by 6.66 times compared with CCT type(TTT,OR=6.66,95%CI=1.59-27.88,P=0.009 7).To conclude,there is a certain degree of linkage disequilibrium between interleukin-1B gene loci rs1143627and rs1143634 in patients with primary frozen shoulder;haplotype TTT formed by these three gene loci may increase the risk of developing primary frozen shoulder.

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 explore the determination method of copper content in Chinese medicinal materials and to provide a scientific basis for the quality control of Chinese medicinal materials.Methods A method for rapid determination of copper content in different varieties of Chinese medicinal materials by X-ray fluorescence spectrometry was established.Results The content of copper in the linear range of 6-40 μg·g-1 showed good linearity with the response strength,which met the quantitative analysis requirements.Conclusion This method has the advantages of being simple,fast,and highly sensitive,and it can be used for rapid quality control of copper content in Chinese medicinal materials.