Extracellular vesicles (EVs) are pivotal mediators of intercellular communication within the tumor immune microenvironment (TME). They are broadly categorized into exosomes, microvesicles, and apoptotic bodies based on their distinct biogenesis pathways. Exosomes originate from the endosomal system via multivesicular body fusion, microvesicles bud directly from the plasma membrane, and apoptotic bodies are released during programmed cell death. By shuttling diverse bioactive cargoes—including proteins, lipids, and nucleic acids such as mRNA, miRNA, and DNA—EVs exert dual modulatory effects on tumor initiation, progression, and immune evasion. Importantly, EVs exhibit remarkable compositional heterogeneity that is intrinsically linked to their cellular origin. Tumor-derived EVs (TDEVs) are typically enriched with immunosuppressive molecules like PD-L1, TGF‑β, and miR-21, which promote tumor immune escape and metastasis. In contrast, EVs derived from immune cells, such as dendritic cells or cytotoxic T lymphocytes, often carry immunostimulatory components including antigens, co-stimulatory molecules, and granzymes, thereby potentiating anti-tumor immunity. This review systematically delineates the biogenesis and molecular composition of EVs, with a particular emphasis on their dynamic regulatory functions within the TME. Specifically, we discuss how EVs mediate intricate crosstalk between immune and tumor cells, facilitating signal transfer that reshapes immune surveillance. For instance, TDEVs can induce macrophage polarization toward an M2-like pro-tumor phenotype, while also suppressing natural killer cell cytotoxicity and dendritic cell maturation. The clinical utility of EV-associated biomarkers in liquid biopsy is increasingly recognized. Circulating EVs carry tumor-specific molecular signatures that mirror the genetic and proteomic alterations of primary tumors, enabling non-invasive early diagnosis, molecular subtyping, and real-time monitoring of therapeutic responses. Their natural biocompatibility, low immunogenicity, and intrinsic ability to traverse biological barriers make them ideal candidates for drug delivery systems. This review explores cutting-edge applications, including the use of EVs in immune checkpoint blockade therapy—for instance, engineered EVs displaying anti-PD-1 antibodies or carrying siRNA to silence immunosuppressive genes. Moreover, EV-based tumor vaccines are being developed, leveraging dendritic cell-derived EVs loaded with tumor antigens to elicit potent T cell responses. The feasibility of loading EVs with therapeutic molecules such as chemotherapeutic agents, oncolytic viruses, or CRISPR-Cas9 components is also under active investigation. The advent of engineered EVs has further expanded their therapeutic potential. Through surface modification or cargo encapsulation, EVs can be tailored for targeted delivery and controlled release, enhancing precision immunotherapy. However, several hurdles impede clinical translation. Current isolation and purification methods, such as ultracentrifugation and size-exclusion chromatography, suffer from low yield and purity. Distinguishing EV subpopulations remains technically challenging due to overlapping size and marker expression. Moreover, the lack of standardized protocols for EV production, characterization, and quality control poses significant barriers to regulatory approval and clinical adoption. Looking forward, the convergence of multi-omics technologies with artificial intelligence offers a powerful approach to decipher EV heterogeneity and identify robust diagnostic signatures. Machine learning algorithms can integrate proteomic, transcriptomic, and lipidomic data from large patient cohorts to construct predictive models for cancer diagnosis and prognosis. Concurrently, advances in bioengineering are enabling the design of next-generation EVs with enhanced targeting specificity, on-demand drug release, and reduced off-target effects. Future efforts should also focus on establishing good manufacturing practice (GMP)‑compliant production processes and conducting rigorous preclinical and clinical evaluations. In summary, this review provides a comprehensive overview of EV biology, their multifaceted roles in the TME, and their transformative potential in cancer diagnostics and therapeutics. By addressing current challenges and leveraging emerging technologies, EV-based strategies are poised to revolutionize precision oncology.

ObjectiveHepatocellular carcinoma (HCC) represents 90% of all primary liver cancers. The main risk factors associated with HCC include viral hepatitis (B and/or C), alcohol abuse, and metabolic dysfunction-associated steatotic liver disease (MASLD), which progressively advance to liver fibrosis, cirrhosis, and ultimately evolve into HCC. Surgical resection represents the most effective treatment for HCC, while recent advances in immunotherapy, including immune checkpoint inhibitors and adoptive cell therapies, have provided improved treatment prospects for patients with unresectable HCC. However, the complex metabolic heterogeneity of HCC limits the therapeutic efficacy. Metabolic intermediates acyl-CoA not only provide energy and substrates for numerous biochemical reactions but also serve as donors for protein lysine acylation, a major class of post-translational modification (PTM). Therefore, a deeper understanding of the molecular mechanisms underlying protein lysine acylation and hepatocarcinogenesis is urgently needed. MethodsThe levels of protein lysine acylation and silence information regulator 5 (SIRT5) expression levels in clinical HCC samples were analyzed by Western blot. Quantitative malonylome and succinylome of HCC samples were analyzed by antibody-based affinity enrichment coupled with tandem mass spectrometry. The proliferation of HCC cells was analyzed with Cell Counting Kit-8 (CCK-8) assays, the apoptosis was quantified by Annexin V-FITC/propidium iodide (PI) staining coupled with flow cytometry, and the ability of cells to migrate was assayed by Transwell assays. The enzymatic activity of glutathione S-transferase Mu 1 (GSTM1) was quantified. Transgenic mice with hepatic overexpression of SIRT5 were constructed using CRISPR-Cas9, and primary hepatocarcinogenesis was induced by administration of diethylnitrosamine. ResultsWestern blot analysis indicated that the expression level of SIRT5 was elevated in clinical samples from HCC patients, and the levels of lysine malonylation, glutarylation, and succinylation were significantly reduced in HCC tissues. Knockout of SIRT5 in MHCC-97H and MHCC-97L hepatoma cells suppressed cell proliferation, and increased the percentage of apoptotic cells significantly. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the differentially malonylome and succinylome of HCC samples revealed significant enrichment in two major classes of biological processes: core energy metabolism (e.g., glycolysis/gluconeogenesis, tricarboxylic acid metabolic process, fatty acid beta oxidation) and detoxification and oxidative stress response (e.g., response to toxic substance, chemical carcinogenesis, reactive oxygen species (ROS)). SIRT5 removes malonylation from lysine residues in GSTM1 and restores its detoxification activity, which is crucial for the survival of hepatocytes under stressed conditions. More importantly, in vivo experiment indicated that hepatic-specific overexpression of SIRT5 in mice accelerated diethylnitrosamine-induced liver fibrosis and hepatocarcinogenesis, indicating the critical role of SIRT5 in HCC progression. ConclusionThis study highlights the previously unrecognized SIRT5-GSTM1 axis as a key regulator in hepatocarcinogenesis, and suggests a potential target for the treatment of patients with HCC.

Aging is the result of the accumulation of various molecular and cellular damages over time,encompassing 12 characteristic hallmarks divided into three categories.These include primary hallmarks such as genomic instability,telomere attrition,epigenetic alterations,loss of proteostasis,and disabled macroautophagy;antagonistic hallmarks like deregulated nutrient sensing,mitochondrial dysfunction,and cellular senescence;and integrative hallmarks including stem cell exhaustion,altered intercellular com-munication,chronic inflammation and dysbiosis.Therefore,investigating cellular signaling factors within a single pathway is insufficient to comprehensively understand the complex mechanisms underlying aging.Casein kinase Ⅱ(CK2),one of the earliest identified protein kinases,is capable of phosphorylating ser-ine/threonine/tyrosine residues on hundreds of substrates.It exhibits highly constitutive expression and activity,and is extensively involved in the regulation of cellular processes such as proliferation,differenti-ation,apoptosis,stress response,metabolism,and immune function.CK2 plays a unique role in coordi-nating the cross-talk and integration of various signaling pathways,which is crucial for maintaining cell survival and homeostasis.Recent studies have revealed that CK2 exhibits perturbations in both expression levels and enzymatic activity during aging process,with notable heterogeneity observed across different animals,tissues and cellular models.Overall,the downregulation of CK2 expression not only promotes the development of primary hallmarks but also alleviates antagonistic hallmarks and ameliorates integrative hallmarks of aging,demonstrating dual effect and interconnected mechanisms.Notably,aberrant activa-tion in CK2 expression and activity are associated with various aging-related diseases,including cancer,cardiovascular diseases,chronic metabolic disorders,neurodegenerative diseases and skeletal degenera-tive conditions.Therefore,maintaining CK2 homeostasis may represent an effective strategy for delaying aging.This review summarizes the latest advances in CK2 and aging research,providing not only deeper insights into the common mechanisms underlying aging and aging-related diseases,but also a theoretical foundation for identifying potential targets for the early prevention of aging-related diseases.

The Lambl's excrescences(LEs)are elongated valvular fronds with excessive mobility,commonly found at the closure lines of heart valves.These filiform projections are thin and long,exhibiting undulating independent motion.LEs predominantly occur on the ventricular side of the aortic valve and on the atrial side of the mitral valve,although there have been documented cases involving tricuspid,pulmonic,and prosthetic valves.To detect LEs,transesophageal echocardiography is often recommended.The high stress experienced by left-sided valves during closure can lead to tears or lesions in the endocardium that initiate LEs pathogenesis.In this particular case study,we present a patient who presented with acute non-ST-segment elevation myocardial infarction alongside LEs located on the non-coronary cusp of the aortic valve.This specific anatomical location posed significant challenges during percutaneous coronary intervention(PCI),as it required direct contact between the guiding catheter and the aortic valve,thereby increasing the risk of dislodging the LEs.Fortunately,our team successfully performed PCI without encountering any notable complications or embolic events.

Objective This study aims to compare the clinical efficacy and complications associated with microwave ablation(MWA)and sublobectomyfor treating stage ⅠA non-small cell lung cancer(NSCLC).Methods Data from stage ⅠA1-ⅠA3 NSCLC patients treated at Yunnan Cancer Hospital between January 2021 and December 2022 were retrospectively collected.A total of 82 patients who underwent MWA and 82 patients who received sublobectomy were selected through propensity score matching.Periprocedural conditions,short-term efficacy,and complications were compared between the two groups.Progression-free survival(PFS)and Disease-free survival(DFS)were monitored and evaluated.The Kaplan-Meier method was employed to construct survival curves,and logistic regression was utilized to analyze the factors influencing complications.Results The MWA group exhibited shorter procedure times,less intraprocedural blood loss,less postprocedure pain,shorter hospitalization durations,and lower hospitalization costs compared to the sublobectomy group(P<0.001).All patients successfully underwent the procedures,with a complete ablation rate of 97.6%and a local control rate of 87.8%in the MWA group.The median follow-up duration was 26.5 months.Survival curve analysis indicated no statistically significant differences in PFS and DFS between the two groups.The primary complications observed were pneumothorax(24.4%,20/82)and moderate to severe pain(11.0%,9/82)in the MWA group,while the sublobectomy group experienced moderate to severe pain(41.5%,34/82)and pulmonary infection(12.2%,10/82).Single-factor analysis demonstrated six variables influenced the occurrence of MWA pneumothorax.Multi-factor logistic regression revealed that the shortest distance from the tumor to the pleura(P=0.021,OR=15.341,95%CI:1.699～24.367)and the number of punctures through the pleura(P=0.024,OR=0.068,95%CI:0.001～0.612)were identified as independent risk factors for pneumothorax.Conclusion MWA demonstrates good efficacy for treating stage ⅠA NSCLC due to minimal trauma,low cost,rapid recovery,few and mild complications.Additionally,MWA and sublobectomy exhibit comparable PFS and DFS,making MWA a safe and effective treatment method.Therefore,MWA is worthy of promotion in clini-cal practice.

Hypercholesterolemia is pathologically characterized by abnormal accumulation of low-density lipoprotein cholesterol,which is closely associated with metabolic dysfunction-associated fatty liver disease and increased cardiovascular risks.Hepatocytes maintain cholesterol homeostasis through LDL receptor-mediated uptake and esterification storage mechanisms.However,chronic cholesterol overload induces mitochondrial dysfunction,reactive oxygen species accumulation,and endoplasmic reticulum stress,leading to hepatocyte injury.Moreover,systemic hypercholesterolemia disrupts gut microbiota balance and impairs short-chain fatty acid and ketone metabolism,exacerbating metabolic disturbances and aggravating hepatic injury through enhanced metabolic stress.In this article,we review the advance of studies on hypercholesterolemia in recent years and summary its association with hepatic injury,which can provide theoretical support for further research.

Numerous studies have shown that depression is main-ly associated with the abnormal expression of connexin 43(Cx43)in astrocytes(Astro)and its mediated dysfunction of gap junction(GJ).However,the molecular mechanism of post-translational modifications targeting Cx43 to regulate neuroin-flammation-associated depression is still unclear.Post-transla-tional modifications of Cx43 mainly include phosphorylation of specific amino acid sites by PKC,PKA,PKG,MAPK and PTK,and protein degradation of Cx43 through the K48/K63 polyubiq-uitylation and deubiquitination pathways,which ultimately lead to protein degradation through K48/K63 polyubiquitination and deubiquitination.These modifications are ultimately involved in the regulation of neuroinflammatory responses through the associ-ation of GJ function.In this paper,we systematically review the role of Cx43 post-translational modifications in neuroinflamma-tion,with the aim of further exploring the potential application of targeting these modifications to modulate the inflammatory re-sponse mechanism in improving depressive symptoms.

Aim To verify the mechanism of action of Carthami Flos-Lycopodii Herba in treating KOA carti-lage inflammation based on network pharmacology and in vitro and in vivo experiments.Methods The effec-tive ingredients of Carthami Flos-Lycopodii Herba were screened through the database,the core targets of"drug disease"were analyzed,and pathway enrichment analy-sis and molecular docking verification were conducted.Experimental verification:Primary chondrocytes were extracted from mice and divided into the control group,IL-1β group,treatment group,and treatment+TLR4 agonist group.CCK-8 method was used to screen the optimal intervention concentration of Carthami Flos-Ly-copodii Herba.ELISA was used to detect the content of inflammatory factors in chondrocytes.Western blot was employed to detect the protein expression related to cellular pathways.Subsequently,a KOA mouse model was constructed using the DMM method.After admin-istration,the knee joint injury of mice was evaluated u-sing safranin O-green staining.ELISA was used to de-tect the levels of inflammatory factors in serum.West-ern blot was employed to detect collagen Ⅱ,MMP13,Aggrecan,and apoptosis related protein expression in cartilage tissue.TUNEL staining was used to detect the apoptosis rate of cells.Results A total of 26 active ingredients of Carthami Flos-Lycopodii Herba were screened,as well as 123 potential targets for treating KOA.The enrichment analysis results indicated that it mainly involved mechanisms such as Toll like receptors and cell apoptosis.The experimental results showed that Carthami Flos-Lycopodii Herba alleviated the in-flammatory response of chondrocytes and affected the expression of pathway related proteins.Compared with KOA mice,safflower stretched muscle grass could im-prove cartilage damage and reduce the concentration of serum inflammatory factors,regulate the expression of collagen Ⅱ,MMP13,Aggrecan,and apoptosis related proteins in cartilage tissue,and reduce the fluorescence intensity of TUNEL staining in the tissue.Conclusions Carthami Flos-Lycopodii Herba can improve KOA cartilage inflammation,and its mechanism may be relat-ed to the TLR4/MyD88/NF-κB signaling pathway.

Objective To explore the feasibility and corresponding implementation methods of constructing a sample resource bank based on individual-level data of completed clinical trials and using it to construct external controls for drug/device clinical trials.Methods Taking the pre-marketing clinical trial of transcatheter active valve replacement(TAVR)for the treatment of aortic valve stenosis as an example,the individual-level databases of multiple trials were standardized to form a sample bank.The original data of any trial in the sample bank were selected as the experimental group,and the remaining samples were selected as the control group.The potential confounding was handled by using the propensity score matching and stratification methods to clarify the process of constructing external controls based on the sample bank of individual-level data of clinical trials.Results This study included individual-level data of single-group trials of 4 TAVR devices,with a total of 569 subjects(59.2%male).The number of subjects in Trials 1 to 4 was 120,120,163,and 166,respectively.Propensity score matching enabled the matching of 113,117,125,and 147 subjects with comparable or similar characteristics from individual-level data from other trials,respectively,demonstrating a high matching success rate.The PS score distribution plot after stratification showed that the proportions of subjects in the experimental and control groups in strata 1 to 5 in scheme 1 were 4/103,11/103,22/92,32/87,and 51/64,respectively.For all constructed external controlled trials,a certain number of control samples with similar baseline characteristics to the experimental groups were distributed within each propensity score stratum.The results of the simulation test also reflected the potential differences between different devices in the 12-month all-cause mortality rate.Conclusions The sample bank constructed with individual-level data from clinical trials,as a high-quality data source,can serve as a source of external control for single-arm trials in the same field,and as a useful supplement to the external control scenario of real-world evidence to support drug and device development.At the same time,targeted research on research methods and bias control measures in related fields is also needed.

Aim To investigate the therapeutic effects of cinnamaldehyde on synovial lesions in mice with knee osteoarthritis(KOA)and its regulatory mecha-nism in the phagocytic function of synovial macropha-ges.Methods In the animal experiments,mouse ser-um and synovial tissue were extracted.HE staining was used to evaluate the inflammatory cell infiltration in the synovial tissue.ELISA was employed to detect the lev-els of inflammatory factors such as interleukins in the serum.Western blot was used to detect the expression of Ras homolog family member A(RhoA),Rho-associ-ated protein kinase 1(ROCK1),myosin light chain(MLC),and p-MLC proteins in the synovial tissue.RT-qPCR was utilized to detect the expression of in-flammatory factors and pathway-related mRNA in the synovial tissue.TUNEL staining was used to detect ap-optosis in the synovial tissue.In the cellular experi-ments,after the intervention,RAW267.4 cells were subjected to Western blot and RT-qPCR for the detec-tion of the aforementioned indicators,and confocal mi-croscopy was used to assess phagocytic function.Re-sults After cinnamaldehyde intervention,the synovial inflammatory infiltration was significantly reduced,the protein and mRNA expression of the RhoA/ROCK1/MLC signaling pathway was markedly downregulated,the fluorescence intensity of TUNEL staining signifi-cantly decreased,and the phagocytic function of macro-phages was enhanced.Conclusion Cinnamaldehyde can inhibit RhoA/Rock1/MLC signaling pathway,en-hance macrophage burial,improve synovial inflamma-tion,and delay the progression of KO A mice.