Single-cell metabolite analysis at the small molecule level reveals intercellular heterogeneity and molecular diversity,especially living cell metabolite analysis which can provide more accurate biochemical information.In this study,a comprehensive single-cell metabolomics mass spectrometry analysis platform was constructed based on continuous ultrasonic sample introduction,aiming to improve the utilization rate of single cells and the efficiency of mass spectrometry detection.This platform utilized mechanical motion generated by a miniaturized ultrasound module,which minimally affected cell integrity and viability,enabling cell suspension and dispersion for up to 60 min,with cell viability exceeding 70%.By comparing cell suspension densities and the cell number of mass spectrometry detections between static and ultrasound groups,the results showed that the ultrasound treatment significantly reduced cell sedimentation rate and increased single-cell mass spectrometry detection efficiency.Applying this platform to single-cell analysis of cell line of mouse cerebellar astrocytes(C8D1A)and mouse glioma(GL261)cells achieved clustering and differential analysis of different cell types,demonstrating the method's potential in analyzing cellular heterogeneity and identifying cells.This approach promised to provide new insights and solutions for single-cell analysis.

Aim To explore the action mechanism of isoquercitrin(IQ)in ameliorating anxiety based on network pharmacology,cellular transcriptomics,molecu-lar docking and animal experiments.Methods The common targets of anxiety disorders and IQ were ob-tained by using relevant databases.The protein-protein interaction network,the biological function and signa-ling pathway enrichment analysis were conducted by u-sing the common targets.Primary hippocampal neurons were cultured in vitro,and corticosterone was added to induce neurons to establish a corticosterone injury mod-el.IQ treatment was added to the culture system,and transcriptomics was used to screen for differentially ex-pressed genes and enrich for differentially expressed pathways.Subsequently,the results were validated by quantitative real-time polymerase chain reaction(qRT-PCR).Possible targets and signaling pathways for IQ treatment on anxiety were speculated and screened u-sing network pharmacology,transcriptomics and molec-ular docking.The anxiety rat model was constructed,and the anxiety state of rats was evaluated after IQ in-tervention,and the protein expression level of hippo-campus was detected to verify the relevant mechanism.Results Network pharmacology,cellular transcrip-tome,and molecular docking analyses revealed that the key mechanism of IQ for anxiety may be related to the BDKRB2/PI3K/Akt signaling pathway.Animal exper-iments showed that IQ was effective in improving anxie-ty behaviour and learning memory ability in rats.IQ increased the movement distance and residence time in the central area of the open field,the time and number percentage of entries into the open arm in the elevated plus maze,and the spontaneous alternations score in the Y maze in rats,and significantly elevated protein expression of BDKRB2,PI3K,Akt and decreased pro-tein expression of NF-κB in the hippocampus.Conclu-sions Isoquercitrin can effectively treat anxiety,and the mechanism of action may be related to the regula-tion of BDKRB2/PI3K/Akt signaling pathway in hip-pocampus.

Objective To systematically evaluate qualitative studies on the experience of inhalation therapy for patients with chronic obstructive pulmonary disease(COPD),in order to provide a basis for healthcare professionals to optimize the care strategies.Methods A systematic search was conducted for qualitative studies on the experience of inhalation therapy for patients with COPD included in domestic and international databases,with a timeframe from the establishment of the database to September 2024.Literature quality was evaluated using the Australian JBI Centre for Evidence-Based Health Care Literature Quality Assessment Criteria for Qualitative Research,and the results were integrated by a pooled integration approach.Results A total of 12 articles were included,and 84 original findings were extracted and summarized into 10 new categories to form 4 integrated results,including low level of drug literacy,differences in perceived efficacy,multiple medication burdens,and multi-dimensional support for inhalation therapy.Conclusion COPD patients have multiple experiences during inhalation therapy.Medical staff should strengthen drug education,enhance patients'cognition and medication skills,foster a positive treatment attitude,and integrate multiple resources to improve patients'inhalation therapy experience.

Aim To explore the action mechanism of isoquercitrin(IQ)in ameliorating anxiety based on network pharmacology,cellular transcriptomics,molecu-lar docking and animal experiments.Methods The common targets of anxiety disorders and IQ were ob-tained by using relevant databases.The protein-protein interaction network,the biological function and signa-ling pathway enrichment analysis were conducted by u-sing the common targets.Primary hippocampal neurons were cultured in vitro,and corticosterone was added to induce neurons to establish a corticosterone injury mod-el.IQ treatment was added to the culture system,and transcriptomics was used to screen for differentially ex-pressed genes and enrich for differentially expressed pathways.Subsequently,the results were validated by quantitative real-time polymerase chain reaction(qRT-PCR).Possible targets and signaling pathways for IQ treatment on anxiety were speculated and screened u-sing network pharmacology,transcriptomics and molec-ular docking.The anxiety rat model was constructed,and the anxiety state of rats was evaluated after IQ in-tervention,and the protein expression level of hippo-campus was detected to verify the relevant mechanism.Results Network pharmacology,cellular transcrip-tome,and molecular docking analyses revealed that the key mechanism of IQ for anxiety may be related to the BDKRB2/PI3K/Akt signaling pathway.Animal exper-iments showed that IQ was effective in improving anxie-ty behaviour and learning memory ability in rats.IQ increased the movement distance and residence time in the central area of the open field,the time and number percentage of entries into the open arm in the elevated plus maze,and the spontaneous alternations score in the Y maze in rats,and significantly elevated protein expression of BDKRB2,PI3K,Akt and decreased pro-tein expression of NF-κB in the hippocampus.Conclu-sions Isoquercitrin can effectively treat anxiety,and the mechanism of action may be related to the regula-tion of BDKRB2/PI3K/Akt signaling pathway in hip-pocampus.

Objective To systematically evaluate qualitative studies on the experience of inhalation therapy for patients with chronic obstructive pulmonary disease(COPD),in order to provide a basis for healthcare professionals to optimize the care strategies.Methods A systematic search was conducted for qualitative studies on the experience of inhalation therapy for patients with COPD included in domestic and international databases,with a timeframe from the establishment of the database to September 2024.Literature quality was evaluated using the Australian JBI Centre for Evidence-Based Health Care Literature Quality Assessment Criteria for Qualitative Research,and the results were integrated by a pooled integration approach.Results A total of 12 articles were included,and 84 original findings were extracted and summarized into 10 new categories to form 4 integrated results,including low level of drug literacy,differences in perceived efficacy,multiple medication burdens,and multi-dimensional support for inhalation therapy.Conclusion COPD patients have multiple experiences during inhalation therapy.Medical staff should strengthen drug education,enhance patients'cognition and medication skills,foster a positive treatment attitude,and integrate multiple resources to improve patients'inhalation therapy experience.

Objective:To systematically evaluate the factors influencing the physical activity levels of patients with chronic obstructive pulmonary disease (COPD) .Methods:Computer retrieval of literature related to the influencing factors of physical activity in patients with COPD in PubMed, Embase, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Wanfang, VIP Database, and China Biology Medicine disc, and the search period was from the establishment of the databases to March 2024. Two researchers independently screened the literature and conducted quality assessment according to inclusion and exclusion criteria. Meta-analysis was conducted using RevMan 5.4 software.Results:A total of 14 studies were included. The results of the Meta-analysis showed that gender [ OR=1.54, 95% CI (1.21, 1.96) ], degree of dyspnea [ OR=1.74, 95% CI (1.37, 2.21) ], fatigue [ OR=2.06, 95% CI (1.48, 2.87) ], and depression [ OR=1.63, 95% CI (1.30, 2.04) ] were influencing factors of physical activity levels in COPD patients ( P<0.05) . Conclusions:The physical activity levels of COPD patients are influenced by gender, degree of dyspnea, fatigue, and depression. Healthcare providers should develop personalized intervention goals and plans to improve patients' participation in and adherence to physical activity.

Objective:To systematically evaluate the factors influencing the physical activity levels of patients with chronic obstructive pulmonary disease (COPD) .Methods:Computer retrieval of literature related to the influencing factors of physical activity in patients with COPD in PubMed, Embase, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Wanfang, VIP Database, and China Biology Medicine disc, and the search period was from the establishment of the databases to March 2024. Two researchers independently screened the literature and conducted quality assessment according to inclusion and exclusion criteria. Meta-analysis was conducted using RevMan 5.4 software.Results:A total of 14 studies were included. The results of the Meta-analysis showed that gender [ OR=1.54, 95% CI (1.21, 1.96) ], degree of dyspnea [ OR=1.74, 95% CI (1.37, 2.21) ], fatigue [ OR=2.06, 95% CI (1.48, 2.87) ], and depression [ OR=1.63, 95% CI (1.30, 2.04) ] were influencing factors of physical activity levels in COPD patients ( P<0.05) . Conclusions:The physical activity levels of COPD patients are influenced by gender, degree of dyspnea, fatigue, and depression. Healthcare providers should develop personalized intervention goals and plans to improve patients' participation in and adherence to physical activity.

The cerebellum is heavily connected with other brain regions, sub-serving not only motor but also nonmotor functions. Genetic mutations leading to cerebellar dysfunction are associated with mental diseases, but cerebellar outputs have not been systematically studied in this context. Here, we present three dimensional distributions of 50,168 target neurons of cerebellar nuclei (CN) from wild-type mice and Nlgn3R451C mutant mice, a mouse model for autism. Our results derived from 36 target nuclei show that the projections from CN to thalamus, midbrain and brainstem are differentially affected by Nlgn3R451C mutation. Importantly, Nlgn3R451C mutation altered the innervation power of CN→zona incerta (ZI) pathway, and chemogenetic inhibition of a neuronal subpopulation in the ZI that receives inputs from the CN rescues social defects in Nlgn3R451C mice. Our study highlights potential role of cerebellar outputs in the pathogenesis of autism and provides potential new therapeutic strategy for this disease.
Animals ; Mice ; Disease Models, Animal ; Cerebellar Nuclei ; Autistic Disorder/pathology* ; Neurons/metabolism* ; Mutation ; Nerve Tissue Proteins/metabolism* ; Male ; Membrane Proteins ; Cell Adhesion Molecules, Neuronal

Objective To investigate the mechanism underlying gasdermin E(GSDME)-mediated pyroptosis in radiation-induced intestinal injury and to find out whether gasdermin(GSDM)family members regulate pyroptosis through similar signaling pathways.Methods Human normal colon epithelial cells(NCM460)and human colon cancer cells(HT-29)were exposed to radiation of different doses and durations before pyroptosis indicators were evaluated by observing pyroptotic bubbles,cell survival,and the cleavage of pyroptosis execution proteins.HT-29 cells overexpressing GSDME were subjected to radiation,followed by enrichment analysis of pyroptosis-related differentially expressed genes using RNA-seq.Results Radiation induced substantial pyroptosis in NCM460 cells.Overexpression of GSDME in HT-29 cells resulted in substantial radiation-induced pyroptosis.The pyroptosis state of human intestinal cells was simulated in the HT-29 model cell line.Overexpressions of GSDME-N and GSDMD-N resulted in the expression of more than 50％ of the differentially expressed genes in the pyroptosis state.Sequencing analysis showed that the genes in the pyroptosis state were mainly overrepresented in immune response,inflammatory response,and Rapl signaling pathway.Conclusion GSDME activation can mediate radiation-induced pyroptosis by producing GSDME-N fragments.GSDM family members participate in pyroptosis in a similar mode of regulation.Furthermore,radiation-induced activation of GSDME/D may regulate pyroptosis through immune response,inflammatory response,and Rap1 signaling pathway.

Hepatocellular carcinoma is one of the most common malignant tumors worldwide, posing a great threat to human health and life. Despite the tremendous progress in understanding the origin and molecular characterization of hepatocellular carcinoma, there are still few therapeutic options that can significantly increase the survival rate and improve the quality of life of patients. Protein post-translational modifications (PTMs) are regulatory mechanisms for protein activity, localization, expression, and interactions with other cellular molecules that induce changes in protein properties and functions. More and more studies have demonstrated that PTMs and immunotherapy play an important role in the development of hepatocellular carcinoma, even in the immunosurveillance of hepatocellular carcinoma and the treatment and prognosis of hepatocellular carcinoma patients. Traditional types of PTMs include phosphorylation, glycosylation, methylation, and ubiquitination. Phosphorylation affects cancer development and progression by regulating tumor cell proliferation, invasion and metastasis, and inhibiting apoptosis. There are two main types of glycosylation: O-glycosylation andN-glycosylation. Abnormal glycosylation not only promotes the proliferation and metastasis of hepatocellular carcinoma cells, but also plays an important role in immune recognition and immune escape. Common methylation modifications include DNA methylation, RNA methylation and histone methylation. Among them, histone methylation, as an important epigenetic regulatory mechanism, is of great theoretical and practical significance for understanding the mechanism of hepatocellular carcinoma as well as carrying out the corresponding prevention and immunotherapy. Ubiquitination plays an important role in the localization, metabolism, function, regulation and degradation of proteins, and it is regulated at different levels by ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2), ubiquitin-conjugating enzyme (E3), and a series of deubiquitinating enzymes (DUBs) and is closely related to hepatocellular carcinoma immunotherapy. This paper begins with a brief overview of the importance of PTMs of proteins, discusses the importance of these traditional types of PTMs in hepatocellular carcinoma immunotherapy, and summarizes the most recent applications of these approaches in hepatocellular carcinoma in order to explore the mechanism of action of PTMs in hepatocellular carcinoma immunotherapy. Then, we summarize the finding that programmed death-ligand 1 (PD-L1) is associated with a variety of conventional types of PTMs, that in-depth study of the mechanisms regulating PD-L1 expression in tumor cells is expected to improve therapeutic efficacy, and that targeting PD-L1 in PTMs is expected to be a new field for exploring hepatocellular carcinoma immunotherapy in the future. Finally, we discuss the current status of research on PTMs for hepatocellular carcinoma immunotherapy and provide new insights and future research directions. In addition to the traditional types of PTMs, multiple novel PTMs have also been identified in published research reports, while the relationship between novel PTMs and hepatocellular carcinoma and the types of PTMs to other undiscovered proteins are still poorly understood, and future research will be focused on a more comprehensive knowledge and understanding of PTMs as well as on exploring new types and mechanisms of PTMs. Overall, further investigation of the role of PTMs in tumor immunity could help to discover new biomarkers and to develop more effective and personalized cancer immunotherapies and targeted therapies, expanding our understanding of cancer biology.