ObjectiveThe exacerbating trend of global population aging poses profound socioeconomic and public health challenges, making the comprehensive elucidation of biological aging mechanisms and the discovery of effective anti-aging interventions an urgent priority in the life sciences. Based on our previous serum metabolomics findings that dimethylglycine, an intermediate metabolite of amino acid metabolism naturally present in the human body, was significantly enriched in the serum of longevity families, this study aimed to systematically investigate the anti-aging effects of dimethylglycine both in living organisms and in controlled laboratory environments, and to preliminarily elucidate its underlying molecular mechanisms. While existing literature indicates that dimethylglycine possesses antioxidant and immunomodulatory properties, its direct anti-aging efficacy and the specific molecular pathways through which it operates remain largely unexplored. MethodsTo comprehensively evaluate the anti-aging properties of dimethylglycine, we utilized replicative senescent human embryonic lung fibroblasts, specifically the WI-38 cell line, as an experimental model in a controlled laboratory environment. Cell viability and safety were thoroughly assessed using Cell Counting Kit-8 and lactate dehydrogenase release assays across various concentrations of dimethylglycine. The impact of dimethylglycine on cellular senescence phenotypes, oxidative stress, and proliferative capacity was evaluated via senescence-associated beta-galactosidase staining, reactive oxygen species fluorescence detection, and 5-ethynyl-2'-deoxyuridine incorporation assays. Furthermore, the molecular alterations of senescence-associated secretory phenotype factors and core senescence signaling pathways were quantified using quantitative reverse transcription polymerase chain reaction for the messenger RNA levels of interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1, and enzyme-linked immunosorbent assay for the measurement of p16 and p21 protein expression levels. For the living organism model, the wild-type nematode Caenorhabditis elegans was used to evaluate systemic physiological effects. We conducted a comprehensive lifespan analysis at 20°C, heat stress resistance survival assays at 35℃, senescence-associated beta-galactosidase staining, lipofuscin accumulation tracking, intracellular reactive oxygen species measurement, and Oil Red O staining to ascertain systemic lipid accumulation. Additionally, network pharmacology bioinformatics tools, including PharmMapper and STRING databases, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were utilized to predict target pathways, alongside highly detailed molecular docking simulations utilizing SwissDock and Protein-Ligand Interaction Profiler to examine interactions with the cytochrome P450 family 2 subfamily C member 9 protein. ResultsThe experimental outcomes robustly demonstrate the potent anti-aging capabilities of dimethylglycine. At the cellular level, toxicity analyses firmly confirmed that dimethylglycine is highly safe; continuous treatment with 50 mol/L and 70 mol/L of dimethylglycine for 5 d did not induce any cellular membrane damage or cytotoxicity, but rather actively promoted cellular proliferation. Utilizing the optimal standardized concentration of 50 mol/L, dimethylglycine treatment significantly ameliorated senescent phenotypic markers in human embryonic lung fibroblasts, which was evidenced by a drastic and highly significant reduction in the senescence-associated beta-galactosidase positive cell percentage (P<0.000 1) and intracellular reactive oxygen species levels (P<0.000 1), alongside a marked increase in the 5-ethynyl-2'-deoxyuridine-positive proliferation rate (P=0.003 5). On a molecular expression scale, dimethylglycine significantly downregulated the messenger RNA expression of multiple core senescence-associated secretory phenotype inflammatory factors, including interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1. Concurrently, it effectively suppressed the protein expression of critical cell cycle arrest markers, diminishing p16 protein levels by 57.3% (P=0.000 4) and p21 protein levels by 27.2% (P=0.000 7). In the nematode Caenorhabditis elegans animal model, dimethylglycine significantly extended the mean lifespan from 20.402 d to an impressive 23.066 d (P<0.000 1) and notably enhanced overall survival rates under severe heat stress environmental conditions (P=0.017). Furthermore, systemic dimethylglycine intervention significantly mitigated age-related physiological decline by decreasing bodily lipofuscin accumulation (P<0.000 1), significantly reducing senescence-associated beta-galactosidase activity, lowering systemic reactive oxygen species fluorescence (P=0.008), and effectively alleviating overall fat accumulation (P<0.000 1). Mechanistically, extensive network pharmacology and Kyoto Encyclopedia of Genes and Genomes analyses strongly revealed that the potential targets of dimethylglycine are significantly enriched in fundamental drug metabolism and oxidative stress response pathways. Precision molecular docking simulations conclusively demonstrated that dimethylglycine forms highly stable structural interactions with the cytochrome P450 family 2 subfamily C member 9 protein, specifically highlighting the definitive formation of 5 stable hydrogen bonds involving serine 365, leucine 366, and serine 429 residues, as well as two critical salt bridge formations with arginine 97 and histidine 368 residues. It is additionally predicted to interact favorably with glutathione S-transferase family proteins. ConclusionDimethylglycine exhibits a profoundly significant and multifaceted anti-aging activity at both the cellular and entire living animal levels. By powerfully alleviating oxidative stress, heavily suppressing the core p16 and p21-dependent cellular senescence signaling pathways, and substantially mitigating the detrimental senescence-associated secretory phenotype, dimethylglycine effectively delays fundamental cellular senescence processes and drastically extends whole-organism lifespan. The biological mechanisms driving these robust protective effects are highly likely closely associated with its direct stable interactions with crucial metabolic and detoxifying enzyme systems, such as cytochrome P450 family 2 subfamily C member 9 and glutathione S-transferase family proteins, thereby systemically improving metabolic dysregulation and restoring critical redox homeostasis. This comprehensive study provides highly solid experimental evidence supporting dimethylglycine as a highly potent and safe potential anti-aging intervention agent, while simultaneously offering a clear molecular mechanistic explanation for the previously documented high abundance of dimethylglycine observed within exceptionally long-lived human populations.

Objective This study reports on the obligatory parasitism of water mites Arrenurus sp. on Anopheles messeae at the Manzhouli Port, Inner Mongolia, China. Methods Duing July 2024, a survey on the mosquito diversity was conducted at the Manzhouli Port. Captured mosquitoes and their ectoparasites were identified to species level. Results A total of 1840 adult mosquitoes were collected, representing species from three genera: Culex(Cx. modestus, Cx. pipiens pallens), Aedes(Ae. dorsalis, Ae. flavidorsalis, Ae. flavescens), and Anopheles (An. messeae). Among all the mosqutioes specimens,3 out of 150 captured An. messeae were found to carry ectoparasitic mites, with number of 2,4,27 mites separately. Morphological and molecular identification reached the same result as water mites(Hydrachnidiae, Hydracrina). COI gene sequence showed 94% similarity with the closest species Arrenurus truncatellus. Conlusions Literature review suggests water mites are host-specific parasitism of mosquito species and herein with the first record of Arrenurus sp. parasiting on An. Messeae in the most high-latitude region globally.

Objective To explore the regulatory effect of methyltransferase-like 3(METTL3)on m6A methylation of Sigma-1 receptor(Sigma-1R)mRNA and mitophagy in cerebral microvascular endothelial cells of sepsis mice.Methods Twelve adult male C57BL/6J mice were randomly divided into the Sham group(without cecal ligation or puncture)and the sepsis group(received cecal ligation and puncture),with 6 mice in each group.m6A methylation levels,and relative expression of METTL3 and Sigma-1R mRNA and protein in cerebral micro-vascular tissues were detected by m6A Dot blot,RT-qPCR,and Western blot,respectively.The bEnd.3 cells were cultured in vitro and divided into the control group(normal culture),model group[lipopolysaccharide(LPS)-induced in vitro sepsis],si-NC group(LPS treat-ment combined with si-NC transfection),and si-METTL3 group(LPS treatment combined with si-METTL3 transfection).m6A Dot blot was used to detect m6A methylation levels of cells in each group;RT-qPCR was used to detect the mRNA expression of METTL3 and Sigma-1R;and Western blot was used to detect the expression of METTL3,Sigma-1R,and autophagy-related proteins of Bnip3,LC3B,and Beclin1.m6A chromatin immunoprecipitation-qPCR was used to detect m6A methylation level of Sigma-1R of cells in each group.JC-1 probe was used to detect the mitochondrial membrane potential,and the ATP assay kit was used to detect the mitochondrial ATP content.Results Compared with the Sham group,the sepsis group showed increased m6A methylation level and METTL3 expression(P<0.05),and decreased Sigma-1R expression in cerebral microvascular tissues(P<0.05).Compared with the control group,m6A methylation level,METTL3 expression and m6A methylation level of Sigma-1R mRNA increased,the expression of Sigma-1R,the normal rate and increase rate of mitochondrial membrane potential,and the ATP content decreased,and the protein expression of Bnip3,LC3B and Beclin1 downregulated in the model group and the si-NC group,with statistically significant differences(P<0.05).Compared with the si-NC group,m6A methylation level,METTL3 expression and m6A methylation level of Sigma-1R mRNA decreased,the expression of Sigma-1R,the normal rate and increase rate of mitochondrial membrane potential elevated,the ATP content increased,and the protein expression of Bnip3,LC3B and Beclin1 upregulated in the si-METTL3 group,with statistically significant differences(P<0.05).Conclusion Silencing METTL3 can inhibit m6A methylation of Sigma-1R mRNA and promote the mitophagy in cerebral microvascular endothelial cells of septic mice.

Objective:The purpose of this study is to explore the influencing factors of poor prognosis in henoch-schonlein purpura(HSP)children,and to construct a nomogram model and verify its validity through the analysis results of Logistic regression model.Methods:Collected the clinical data of 170 HSP children who were treated in Fuzhou Luoyuan County Hospital from January 2015 to May 2023 were retrospectively analyzed.They were divided into good prognosis group and poor prognosis group according to the prognosis after treatment.The clinical data and related biochemical indexes of different prognostic groups were compared.The factors of poor prognosis in HSP children were analyzed by Logistic regression model.A predictive model(nomogram)for poor prognosis in HSP children were constructed and evaluated its predictive performance.Prediction accuracy were evaluateed by receiver operating characteristic(ROC)curve.Results:170 HSP children,69 cases(40.59％)had poor prognosis and 101 cases(59.41％)had good prognosis.C-reactive protein(CRP),kidney injury at initial onset,platelet count(PLT),respiratory infection,recurrent rash,and immunoglobulin A(IgA)levels in poor prognosis group were higher than those in good prognosis group(P＜0.05).Multivariate logistic analysis showed that,recurrent rash,respiratory infection,elevated PLT,CRP and IgA levels were independent influencing factors of poor prognosis in HSP children(P＜0.05).The consistency index(C-index)of the column chart model established based on the results of multiple factor analysis was 0.798.The internal verification was carried out by Bootstrap self-sampling method,and the average absolute error of calibration curve was 0.017.ROC curve was drawn according to independent influencing factors and nomogram prediction probability,the area under the curve was 0.674(recurrent rash),0.649(respiratory infection),0.777(PLT),0.733(CRP),0.749(IgA)and 0.910(nomogram prediction probability)respectively.Conclusion:Recurrent rash,respiratory infection,PLT,CRP and IgA are independent factors affecting the prognosis of HSP children.The column chart prediction model constructed based on the above factors has certain predictive value for the poor prognosis in HSP children.

Aim To explore the effect of astragalosideⅣ(AS-Ⅳ)on lipopolysaccharide(LPS)-induced po-larization and migration of RAW 264.7 macrophages and the underlying mechanism.Methods 1 mg·L-1 LPS was used to construct cell migration model.Scratch assay was utilized to determine cell migration rate.Immunofluorescence staining was utilized to de-tect the expression and location of F4/80,iNOS and Arg-1.CCK-8 assay was used to determine the viabili-ty of RAW 264.7 cells.Griess assay was used to measure NO content.Molecular docking was used to analyze the interaction between AS-Ⅳ and the core tar-gets such as cGAS and STING protein.Western blot was employed to detect the expression of iNOS,Arg-1,cGAS,STING,NF-κB p65 and p-NF-κB p65 protein.Results AS-Ⅳ significantly inhibited the migration and M1 polarization of RAW 264.7 cells induced by LPS.Moreover,AS-Ⅳ could interact with cGAS and STING protein,especially cGAS.Further Western blot assay showed that AS-Ⅳ significantly downregulated the expression of iNOS,cGAS,STING and p-NF-κB p65 protein.Conclusions AS-Ⅳ could promote mac-rophage M1 to M2 polarization,thereby inhibited mac-rophage migration through restraining the cGAS/STING/NF-κB signaling pathway,which provides a new therapeutic target for AS-Ⅳ to improve the early inflammatory response of AS.

Aim To investigate the mechanism of 8-hydroxygenistein(8-OHG)in mitigating high-altitude induced heart injury(HAHI)via network pharmacolo-gy,molecular docking and animal experiment.Meth-ods 8-OHG-related targets were obtained from Swis-sTargetPrediction,Similarity ensemble approach,Su-perPred and PharmMapper databases.Genecards and OMIM databases were utilized for retrieving HAHI-re-lated targets.Venn diagram was drawn using R pack-age.STRING 11.5 and Cytoscape 3.9.1 were used to construct the protein-protein interaction network and screen core targets.GO and KEGG enrichment analysis were carried out using DAVID database.AutoDock Vi-na software was used for molecular docking.Visualiza-tion was performed using PyMOL 3.0.0 software.The HAHI model was established,and the the mice were randomly divided into the control group,model group and 8-OHG group.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of myo-cardial tissue.Western blot was applied for detecting the expression levels of related proteins in myocardial tissue.Results A total of 73 overlapping targets be-tween 8-OHG and HAHI were screened,with ALB,AKT1,ESR1,HSP90AA1,NFKB1 and MMP9 were regarded as core targets.Molecular docking results in-dicated that 8-OHG had strong binding ability with these core targets.GO functional enrichment analysis obtained 185 biological processes,including negative regulation of apoptosis,response to hypoxia and in-flammatory response,38 cell compositions,including cytosol,cytoplasm,plasma membrane,as well as 71 molecular functions,including protein binding,metal ion binding,enzyme binding and so on.Altogether 55 signaling pathways were identified via KEGG enrich-ment analysis,including PI3 K/Akt signaling pathway,HIF-1 signaling pathway and MAPK signaling pathway.The results of animal experiments showed that 8-OHG could significantly improve the myocardial histopatho-logical change induced by high-altitude hypoxia expo-sure.Western blot results showed that compared with the normal group,the ratio of p-PI3K/PI3K and p-Akt/Akt in the myocardial tissue of mice in the model group significantly decreased,while the protein expres-sion of Beclin-1 and the ratio of LC3B-Ⅱ/LC3B-Ⅰsignificantly increased,while 8-OHG could reverse these changes.Conclusion The mechanism of 8-OHG in alleviating HAHI is related to its activation of PI3K/Akt signaling pathway,thereby inhibiting auto-phagy induced by high-altitude hypoxia exposure.

Objective To compare the accuracy and clinical application value of contrast-induced fractional flow reserve(cFFR),resting full-cycle ratio(RFR),and quantitative flow ratio(QFR)in evaluating coronary artery borderline lesions,using fractional flow reserve(FFR)as the gold standard.Methods A retrospective study was conducted including 143 patients with 143 lesions who underwent coronary angiography(CAG)and were tested for cFFR,RFR,QFR,and FFR at Peking University Third Hospital from September 2020 to January 2022.Clinical data,CAG lesion anatomical data,and target vessel cFFR,RFR,QFR,and FFR measurements were collected.The correlation and diagnostic concordance of cFFR,RFR,QFR,and FFR were analyzed.Results The mean age of the 143 patients was 66(58,71)years,with 90(62.9％)being male.Among them,60(42.0％)patients were diagnosed with unstable angina,and 115(80.4％)target lesions were located in the left anterior descending artery.Correlation analysis showed that RFR,QFR,and cFFR were all significantly correlated with FFR(r=0.956,r=0.861,r=0.751,P＜0.001).Bland-Altman analysis demonstrated high agreement between cFFR,RFR,QFR,and FFR.Receiver operating characteristics(ROC)curve analysis showed that the area under the curve(AUC)for RFR corresponding to FFR ≤0.80 was 0.92(95％CI 0.87-0.96),for QFR was 0.89(95％CI 0.83-0.95),and for cFFR was 0.96(95％CI 0.94-0.99).Conclusions cFFR,RFR,and QFR show high concordance with FFR,with similar diagnostic consistency between the three methods and FFR.

Patient-Reported Outcomes(PRO)is a subjective outcome indicator based on the concept of"patient-centered",in which patients report their own information such as health status,functional status,and treatment feelings,which is an ef-fective supplement to traditional objective outcome indicators.Based on the development status of the Patient Reported Outcome Measure(PROM),it systematically analyzed the concepts related to PRO with the goal of clarifying the Patient Experience Data(PED)with research value,and proposed six core types of PROs in the medical field,including health-related quality of life,health preference,health behavior,symptom burden,functional status and patient-reported experience.For each type of PRO,the key characteristics of the study were analyzed,the evidence and application directions of related PED and the production based on PED were described,and the existing international mainstream PROMs were reviewed.It is hoped that the connotation of PRO-related concepts can be comprehensively expounded,and PEDs with research significance and value can be explored,so as to provide guidance for the focus of PRO-related research in China,so as to promote the application and development of PRO in China.

Patient-Reported Outcomes(PRO)refers to a subjective outcome measure based on the concept of"patient-cen-tered",in which patients report their own health status,functional status,treatment feelings and other information,which is an ef-fective supplement to traditional objective outcome indicators.With the continuous promotion of PRO research,its application in the world is also constantly enriched.It takes the different application scenarios of PRO as the starting point and the evidence-based evidence that PRO can generate as the evaluation object,reviews the application status of PRO in the US,the European Union,the UK,Canada and other countries,regions or organizations,and summarizes the main application methods of PRO in different medical decision-making scenarios,in order to provide international experiences for the application research of PRO in China.

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.