This study was aimed at investigating the effects of demethylase fat mass and obesity-associated protein(FTO)and methyltransferase methyltransferase like protein 3(METTL3),key molecules in N6-methyladenosine(m6A)modification,on the replication and proliferation of Japanese encephalitis virus(JEV).Recombinant lentiviruses were generated by packaging the FTO and green fluorescent protein into lentiviral vectors.Neuro2a cells,a mouse neuroblastoma cell line,were infected with the lentivirus,and stable FTO-expressing cell lines were obtained through puromycin selection.Successful overexpression of FTO was confirmed through fluorescence microscopy,real-time quantitative PCR,and western blot analysis.When Neuro2a cells overexpressing FTO were infected with JEV,the overexpression of FTO decreased JEV replication in the cells,and increased the expression of interferon(IFN)and related molecules.Additionally,treatment of JEV-infected Neuro2a cells with the METTL3-specific inhibitor STM2457 resulted in a dose-dependent decrease in JEV replication and viral protein expression.These findings suggested that lowering m6A methylation levels inhibits JEV replication,thus shedding light on the regulatory role of methylation modification in JEV replication.

Objective To study NF1 gene variations in 10 suspected cases of neurofibromatosis type I(NF1)and their parents,thereby providing a basis for genetic counselling,clinical diagnosis,and treatment of this disease.Methods A total of 10 patients diagnosed with,or suspected of having NF1 at Northwest Women's and Children's Hospital from March to December 2023 were selected as study subjects.Whole exome sequencing(WES)was performed to analyse NF1 gene mutations in the patients and their parents,with the findings validated by Sanger sequencing.Results Pathogenic mutations were identified in 6 of the 10 families,with the remaining 4 cases showing no pathogenic gene mutations.In Family 1,a de novo mutation,c.6505A>T,was detected in the NF1 gene.In Family 2,a de novo mutation,c.6705-3C>A,was identified in the NF1 gene.In Family 3,a de novo mutation,c.6853delT,was detected in the NF1 gene.In Family 4,a de novo mutation,c.2446C>T,was found in the NF1 gene.In Family 5,a paternal mutation,c.6067T>A,was identified in the NF1 gene.In Family 6,a paternal mutation,c.2991_2993dup,was detected in the NF1 gene.Conclusion The identification of new mutation sites enriches the mutation spectrum of the NF1 gene.This study provides important guidance for genetic counselling and prenatal diagnosis,offering crucial information for families with reproductive needs.

Objective To investigate the relationship between serum levels of micro RNA(miR)-497-5p,fibroblast growth factor-2(FGF2)and brain-derived neurotrophic factor(BDNF),with cognitive dysfunction in Parkinson's disease(PD)patients.Methods From April 2022 to April 2024,86 PD patients(study group)treated in Wuhan Xinzhou District People's Hospital and 60 healthy individuals(control group)who underwent physical examination in Wuhan Xinzhou District People's Hospital were selected.Serum miR-497-5p levels were determined by real-time quantitative polymerase chain reaction(RT-qPCR),and serum FGF2 and BDNF levels were measured by enzyme linked immunosorbent assay(ELISA).Spearman was applied to analyze the correlation between serum miR-497-5p,FGF2,BDNF levels and montreal cognitive assessment(MoCA score).Logistic analysis was applied to analyze the factors influencing cognitive dysfunction in PD patients.Receiver operating characteristic(ROC)was applied to analyze the diagnostic value of serum miR-497-5p,FGF2 and BDNF for cognitive dysfunction in PD patients.Results The serum miR-497-5p(2.73±0.67)expression level in the study group was obviously increased than that in the control group(1.04±0.34),while the expression levels of FGF2(1.94±0.45ng/ml)and BDNF(8.31±2.44ng/ml)were obviously reduced than those in the control group(2.71±0.69ng/ml,12.81±3.07ng/ml),with significate differences(t=17.977,8.161,9.850,all P<0.05).Serum miR-497-5p was negatively correlated with MoCA score(r=-0.331,P<0.05),while FGF2 and BDNF levels were positively correlated with MoCA score(r=0.404,0.361,all P<0.05).Cognitive dysfunction group had significantly higher disease duration,FGF2,BDNF levels,and MoCA scores than the cognitively normal group.The course of disease and miR-497-5p levels in the cognitive dysfunction group were obviously higher than that in the normal cognitive function group,FGF2,BDNF levels,and MoCA scores in the cognitive dysfunction group were obviously lower than that in the normal cognitive function group(t=2.350～11.792,all P<0.05).The course of disease and serum miR-497-5p were risk factors for cognitive dysfunction in PD patients(Wald χ2=4.712,5.704,all P<0.05),while MoCA score,serum FGF2,and BDNF were protective factors for cognitive dysfunction in PD patients(Wald χ2=4.499,5.556,5.217,all P<0.05).The AUC and sensitivity of the combination of serum miR-497-5p,FGF2,and BDNF in PD patients with cognitive impairment were higher than those of individual diagnosis.The diagnostic effect of the combination of miR-497-5p,FGF2,and BDNF in PD patients with cognitive dysfunction was better than that of individual diagnosis,and the differences were statistically significant(Z=2.279,2.236,2.123,all P<0.05).Conclusion Elevated serum miR-497-5p level and decreased FGF2 and BDNF levels can increase the risk of cognitive dysfunction in PD patients,and the combination of the three has good diagnostic value for cognitive dysfunction in PD patients.

Objective:To construct an integrated management model for early screening and diagnosis of PCa based on the In-novative Care for Chronic Conditions Framework(ICCC)and the 1+1 contract-based tiered diagnosis and treatment system(TDTS)in China.Methods:Based on the 1+1 contract-based TDTS platform,we conducted PCa screening for the male residents aged 60 years and above during health check-ups in Pujin Community Health Center from January 1,2023 to December 31,2023.For those with abnormal total prostate-specific antigen(tPSA)≥4 μg/L,we promptly referred them to higher-level hospitals for further diagno-sis and treatment via the two-way referral green channel platform and information sharing service using the 1+1 contract model.We further analyzed the relevant data on screening and diagnosis.Results:A total of 4 080 males aged 71.39±5.059 years received PCa screening from January to December 2023.PSA screening was performed in 43.96％of the male residents,revealing 654 cases of PSA abnormality,with a PSA positivity rate of 16.03％,which was higher than that found in the previous large-scale PCa screenings in other regions of China.Among the males with PSA abnormality,292(44.65％)expressed their willingness for medical referral,while the others did not seek further medical attention for reasons of being asymptomatic,low awareness of the disease,no accompany for medical visits,and concerns about further costs of diagnosis and treatment.Prostate biopsy was recommended to 154 cases after further examinations,which was accepted by 92(59.74％).Fifty-eight cases were diagnosed with Pa,and thedetection rate reached 63.04％.Conclusion:The integrated management model for PSA examination-based early screening and diagnosis of PCa using the 1+1 contract-based TDTS platform is plays a significant role in enhancing peoples awareness and knowledge of PCa and improving the early detection rate of the malignancy.

Objective To develop a nomogram-based predictive model for assessing the risk of respiratory distress syndrome(RDS)in premature infants in the high-altitude region of Dali.The predictive performance and clinical applicability of the model will be systematically evaluated to provide evidence-based guidance for the early diagnosis and clinical management of respiratory distress in premature infants.Methods A total of 680 preterm infants admitted to the Dali Maternal and Child Health Hospital between January 2020 and December 2024 were enrolled in the study and randomly divided into a training set(n=476)and a validation set(n=204)at a ratio of 7∶3.Independent predictors were identified through univariate logistic regression and multivariate stepwise regression analyses,and a nomogram model was subsequently developed using R software.The performance of the model,including its discrimination,calibration,stability,and clinical applicability,was evaluated using the receiver operating characteristic curve(ROC),Hosmer-Lemeshow goodness-of-fit test,bootstrap resampling method,and decision curve analysis(DCA).Results The final model incorporated seven independent variables:gestational age,birth weight,Apgar score,blood oxygen saturation,gestational hyperglycemia,prenatal glucocor-ticoid therapy,and maternal history of infection.The areas under the curve(AUCs)for the training and validation sets were 0.88(95%CI:0.84～0.92)and 0.83(95%CI:0.76～0.89),respectively,with all Hosmer-Lemeshow test p-values exceeding 0.05.The bootstrap-corrected AUC was 0.85(95%CI:0.81～0.89).DCA indicated that the model achieved the highest net benefit at a risk threshold range of 10%to 35%.Conclusions This model integrates multiple risk factors associated with the occurrence of RDS in plateau environments,demonstrating robust predictive performance for RDS in preterm infants residing in high-altitude areas such as Dali.It can serve as a valuable tool for risk stratification and clinical decision-making,and may also provide a reference for future multicenter prospective studies.

The European Society of Cardiology (ESC) released the "2024 ESC guidelines for the management of elevated blood pressure and hypertension" on August 30, 2024. This guideline updates the 2018 "Guidelines for the management of arterial hypertension." One notable update is the introduction of the concept of "elevated blood pressure" (120-139/70-89 mm Hg). Additionally, a new systolic blood pressure target range of 120-129 mm Hg has been proposed for most patients receiving antihypertensive treatment. The guideline also includes numerous additions or revisions in areas such as non-pharmacological interventions and device-based treatments for hypertension. This article interprets the guideline's recommendations on definition and classification of elevated blood pressure and hypertension, and cardiovascular disease risk assessment, diagnosing hypertension and investigating underlying causes, preventing and treating elevated blood pressure and hypertension. We provide a comparison interpretation with the 2018 "Guidelines for the management of arterial hypertension" and the "2017 ACC/AHA guideline on the prevention, detection, evaluation, and management of high blood pressure in adults."

This study investigated the therapeutic effects and mechanisms of Modified Yiyi Fuzi Baijiang Powder on ulcerative colitis(UC) in rats from the perspective of dampness. SD rats were randomly allocated into six groups(n=10): control, model, mesalazine, and Modified Yiyi Fuzi Baijiang Powder at low(3.96 g·kg~(-1)·d~(-1)), medium(7.92 g·kg~(-1)·d~(-1)), and high(15.84 g·kg~(-1)·d~(-1)) doses. UC was induced in all groups except the control by administration with 3% dextran sulfate sodium(DSS) solution for 7 days. The disease activity index(DAI) was recorded, and the colon tissue was collected for analysis. Histopathological changes were assessed by hematoxylin-eosin staining. Serum levels of D-lactic acid(D-LA) and diamine oxidase(DAO) were measured by ELISA. Immunohistochemistry and PCR were employed to evaluate the expression of aquaporins(AQP3, AQP4) and tight junction proteins [zonula occludens-1(ZO-1) and occludin] at both protein and mRNA levels. Compared with the control group, the model group showed an increased DAI scores(P<0.05), intestinal mucosal damage, elevated serum levels of DAO and D-LA(P<0.05), and decreased expression of AQP3, AQP4, ZO-1, and occludin(P<0.05). Treatment with Modified Yiyi Fuzi Baijiang Powder reduced the DAI scores(P<0.05), lowered the serum levels of D-LA and DAO(P<0.05), and upregulated the expression of AQP3, AQP4, ZO-1, and occludin at both protein and mRNA levels compared with the model group. These findings suggest that Modified Yiyi Fuzi Baijiang Powder exerts therapeutic effects on UC by reducing the intestinal mucosal permeability, promoting colonic mucosal repair, and regulating abnormal intestinal water metabolism, which may involve the upregulation of AQP3 and AQP4 expression.
Animals ; Colitis, Ulcerative/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Rats ; Intestinal Mucosa/metabolism* ; Male ; Aquaporin 3/metabolism* ; Aquaporin 4/metabolism* ; Permeability/drug effects* ; Humans ; Powders ; Intestinal Barrier Function

Malignant proliferating liver cancer cells possess the ability to detect and respond to various body signals, thereby facilitating tumor growth, invasion, and metastasis. One crucial mechanism through which hepatocellular carcinoma (HCC) cells interpret these signals is crosstalk. Within liver cancer tissues, cancer cells engage in communication with hepatic stellate cells (HSCs), tumor-associated macrophages (TAMs), and immune cells. This interaction plays a pivotal role in regulating the proliferation, invasion, and metastasis of HCC cells. Crosstalk occurs in multiple ways, each characterized by distinct functions. Its molecular mechanisms primarily involve regulating immune cell functions through the expression of specific receptors, such as CD24 and CD47, modulating cell functions by secreting cytokines like transforming growth factor-β (TGF-β) and platelet-derived growth factor (PDGF), and mediating cell growth and proliferation by activating pathways such as Wnt/β-catenin and Hedgehog. A comprehensive understanding of the mechanisms and interactions within crosstalk is essential for unraveling the pathogenesis of HCC. It also opens up new avenues for the development of innovative therapeutic strategies. This article reviews the relationship between crosstalk and the progression of HCC, offering insights and inspiration for future research.
Humans ; Carcinoma, Hepatocellular/metabolism* ; Liver Neoplasms/metabolism* ; Hepatic Stellate Cells/physiology* ; Disease Progression ; Signal Transduction/physiology* ; Transforming Growth Factor beta/metabolism* ; Cell Proliferation ; Hedgehog Proteins/metabolism* ; Tumor-Associated Macrophages ; Platelet-Derived Growth Factor/metabolism* ; Cell Communication/physiology*

Aim To elucidate the molecular mecha-nism underlying the inhibitory effect of liquidambaric acid(LDA)targeting TNF receptor associated factor 6(TRAF6)in colorectal cancer.Methods This study employed microscale thermophoresis(MST),drug af-finity responsive target stability assay(DARTS)and cellular thermal shift assay(CETSA)to confirm the direct binding of LDA to TRAF6.Additionally,we generated TRAF6 knockout colorectal cancer HCT116 cells using CRISPR/Cas9 technology,and assessed the impact of LDA on TRAF6-regulated Hippo/YAP and Wnt signaling pathways through immunofluorescence a-nalysis and TOPFlash/Renilla luciferase reporter sys-tem.Co-IP and proximity ligation assays(PLA)were conducted to investigate LDA-regulated TRAF6 pro-tein-protein interactions and elucidate molecular mech-anisms.Results The direct binding of LDA to TRAF6 was confirmed in cell lysates and living cells.LDA promoted TRAF6-dependent nuclear translocation of YAP in colorectal cancer cells,and inhibited Wnt signaling by overexpressing TRAF6.Co-IP and PLA revealed that TRAF6 formed a tripartite complex with YAP and β-catenin in colon cancer cells,where TRAF6 was a key scaffolding protein of the tripartite complex.LDA disrupted the interactions between the TRAF domain of TRAF6 and YAP,as well as YAP and β-catenin.Conclusion LDA regulates Hippo/YAP signaling pathway by targeting TRAF6 and inhib-its colorectal cancer.