Objective To investigate the association between physical activity levels and blood lipids among college freshmen, and to provide scientific evidence for the health management of college freshmen. Methods An electronic questionnaire survey on physical activity was conducted on freshmen of a university, and fasting blood biochemical indicators were detected. The International Physical Activity Questionnaire (IPAQ) short form was used to evaluate the physical activity levels of the participants. Dyslipidemia was defined as an abnormality in any one of the following serum lipid parameters: total cholesterol, triglycerides, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), or non-high-density lipoprotein cholesterol. Binary logistic regression and stratified analyses were employed to explore the relationship between physical activity and blood lipids. Results A total of 3 401 participants were included, with an average age of 18.45 ± 0.92 years, and 60.5% were female. The prevalence of dyslipidemia was 17.7%, with a higher rate among males (22.1%) than females (14.8%). After adjusting for confounding factors related to blood lipids, high-intensity physical activity was negatively associated with the risk of elevated LDL-C among males (OR = 0.36, 95% CI: 0.13–0.99, P = 0.049). Conclusion Among freshmen at a medical college in Hubei Province, high-intensity physical activity is negatively associated with the risk of elevated LDL-C in males, but this association needs to be further confirmed by larger prospective cohort studies.

This study aimed to explore the molecular mechanism of rubioncolin C(RuC) in inhibiting gastric cancer(GC). AGS and MGC803 cell lines were selected as cellular models. After treating the cells with RuC at different concentrations, the effects of RuC on the proliferation ability of GC cells were assessed using the CCK-8 method, real-time cellular analysis(RTCA), and colony formation assays. Transmission electron microscopy was used to observe subcellular structural changes. Immunofluorescence was applied to detect LC3 fluorescent foci. Acridine orange staining was used to evaluate the state of intracellular lysosomes. Western blot was employed to detect the expression of autophagy-related proteins LC3Ⅱ, P62, and lysosomal cathepsin D(CTSD). The SuperPred online tool was used to predict the target proteins that bound to RuC, and molecular docking analysis was conducted to identify the interaction sites between RuC and CTSD. The drug affinity responsive target stability(DARTS) assay was performed to detect the direct binding interaction between RuC and CTSD. The results showed that RuC significantly inhibited the proliferation and colony formation of GC cells at low concentrations, with 24-hour half-maximal inhibitory concentrations(IC_(50)) of 3.422 and 2.697 μmol·L~(-1) for AGS and MGC803 cells, respectively. After 24 hours of treatment with RuC at concentrations of 1, 2, and 3 μmol·L~(-1), the colony formation rates for AGS cells were 61.0%±1.5%, 28.0%±0.5%, and 18.2%±0.5%, respectively, while the rates for MGC803 cells were 56.0%±0.5%, 23.3%±1.0%, and 11.8%±1.0%, all of which were significantly reduced. Transmission electron microscopy revealed that RuC promoted an increase in autophagosome formation in GC cells. Immunofluorescence detection showed that LC3 fluorescent foci of GC cells increased with the increase in RuC dose. RuC up-regulated the expression of autophagy-related proteins LC3Ⅱ and P62 in GC cells. Acridine orange staining indicated that RuC altered the acidic environment of lysosomes. SuperPred online prediction identified CTSD as a potential target protein of RuC. Western blot analysis revealed that RuC induced the up-regulation of the inactive precursor of CTSD in GC cells. CTSD activity assays indicated that RuC reduced the activity of CTSD. Molecular docking simulations found that RuC bound to the substrate-binding region of CTSD, forming hydrogen bonds with the Tyr205 and Asp231 residues. Microscale thermophoresis and DARTS assays further confirmed that RuC directly bound to CTSD. In summary, RuC inhibits lysosomal activity by targeting and down-regulating the expression of CTSD, thereby inducing autophagosome accumulation in GC cells.
Humans ; Stomach Neoplasms/enzymology* ; Cathepsin D/chemistry* ; Cell Line, Tumor ; Molecular Docking Simulation ; Cell Proliferation/drug effects* ; Autophagosomes/metabolism* ; Autophagy/drug effects*

Objective To predict the lymph node metastasis status of patients with invasive pulmonary adenocarcinoma by constructing machine learning models based on primary tumor radiomics, peritumoral radiomics, and habitat radiomics, and to evaluate the predictive performance and generalization ability of different imaging features. Methods A retrospective analysis was performed on the clinical data of 1 263 patients with invasive pulmonary adenocarcinoma who underwent surgery at the Department of Thoracic Surgery, Jiangsu Province Hospital, from 2016 to 2019. Habitat regions were delineated by applying K-means clustering (average cluster number of 2) to the grayscale values of CT images. The peritumoral region was defined as a uniformly expanded area of 3 mm around the primary tumor. The primary tumor region was automatically segmented using V-net combined with manual correction and annotation. Subsequently, radiomics features were extracted based on these regions, and stacked machine learning models were constructed. Model performance was evaluated on the training, testing, and internal validation sets using the area under the receiver operating characteristic curve (AUC), F1 score, recall, and precision. Results After excluding patients who did not meet the screening criteria, a total of 651 patients were included. The training set consisted of 468 patients (181 males, 287 females) with an average age of (58.39±11.23) years, ranging from 29 to 78 years, the testing set included 140 patients (56 males, 84 females) with an average age of (58.81±10.70) years, ranging from 34 to 82 years, and the internal validation set comprised 43 patients (14 males, 29 females) with an average age of (60.16±10.68) years, ranging from 29 to 78 years. Although the habitat radiomics model did not show the optimal performance in the training set, it exhibited superior performance in the internal validation set, with an AUC of 0.952 [95%CI (0.87, 1.00)], an F1 score of 84.62%, and a precision-recall AUC of 0.892, outperforming the models based on the primary tumor and peritumoral regions. Conclusion The model constructed based on habitat radiomics demonstrated superior performance in the internal validation set, suggesting its potential for better generalization ability and clinical application in predicting lymph node metastasis status in pulmonary adenocarcinoma.

The nucleotide binding oligomerization domain like receptor protein 3(NLRP3)inflamma-some is an intracellular polymeric protein complex that plays an important role in inflammation and immune response.In recent years,the study of NLRP3 inflammasome in hematological malignancies has become a hot topic.Leukemia is a large group of clonal hematologic malignancies that affect the maturation and/or prolifer-ation of bone marrow or lymphocytes.More and more studies have shown that NLRP3 inflammatory micro-molecules are involved in the occurrence and development of leukemia,and it is expected to become an impor-tant therapeutic target for leukemia in the future.In this review,the structure,biological function,activation pathway of NLRP3 inflammasome and its relationship with different types of leukemia were reviewed,and its potential application in the treatment of leukemia was discussed.

ObjectiveTo elucidate the potential mechanisms by which the classic prescription Anmeidan alleviates cognitive impairment in insomnia model rats through metabolic profiling. MethodsA total of 60 SD rats were randomly divided into six groups： blank group， model group， low-， medium-， and high-dose Anmeidan groups， and the Suvorexant group， with 10 rats in each group. Except for the blank group， the insomnia model was established in all other groups via intraperitoneal injection of para-chlorophenylalanine. The Suvorexant group was administered Suvorexant solution （30 mg·kg^-1·d^-1） by gavage， while the low-， medium-， and high-dose Anmeidan groups received Anmeidan decoction （4.55， 9.09， 18.18 g·kg^-1·d^-1） by gavage. The blank group received an equivalent volume of normal saline. The open field test was used to assess spatial exploration and anxiety/depressive-like behaviors in rats. Serum levels of epidermal growth factor （EGF）， brain-derived neurotrophic factor （BDNF）， and vasoactive intestinal peptide （VIP） were measured using enzyme-linked immunosorbent assay （ELISA）. Untargeted metabolomics was employed to identify differential metabolites in rat serum， and systematic biological methods were applied to analyze the potential targets and pathways of Anmeidan. ResultsCompared to the blank group， the model group exhibited significant reductions in total distance traveled， average speed， number of entries into the central area， time spent in the central area， and frequency of upright events （P<0.01）， along with significant decreases in VIP， EGF， and BDNF levels （P<0.05，P<0.01）. A total of 100 differential metabolites were identified between the model and blank groups. Compared to the model group， the low-， medium-， and high-dose Anmeidan groups showed significant increases in total distance traveled， average speed， number of entries into the central area， time spent in the central area， and frequency of upright events （P<0.05，P<0.01）， as well as a significant increase in VIP levels （P<0.05，P<0.01）. Anmeidan significantly reversed abnormal changes in 67 metabolites compared to the model group. A combined analysis identified 134 potential targets of Anmeidan， with network topology analysis suggesting that Caspase-3， B-cell lymphoma 2 （Bcl-2）， nuclear transcription factor-κB （NF-κB）， interleukin-1β （IL-1β）， interleukin-2 （IL-2）， matrix metalloproteinase-9 （MMP-9）， and Toll-like receptor 4 （TLR4）， among others， may serve as key targets of Anmeidan. Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis revealed major enriched pathways， including the cyclic adenosine monophosphate （cAMP） signaling pathway， hypoxia inducible factor-1 （HIF-1） signaling pathway， and IL-17 signaling pathway. ConclusionThis study demonstrates that Anmeidan can recalibrate abnormal metabolic profiles in insomnia model rats to mitigate cognitive impairment， with its mechanisms of action potentially involving the regulation of immune-inflammatory responses， energy metabolism， and apoptosis-related pathways.

AIM: To investigate the mechanism of Hexue Mingmu Tablets(HXMMT)in improving diabetic retinopathy(DR)based on transcriptomics.METHODS: Zebrafish DR models were established by 3-day glucose induction(130 mmol/L)starting at 3 days post-fertilization(dpf). Larvae were randomized into four groups: control group(CG; aquaculture water), model group(MG; 130 mmol/L glucose), low-dose HXMMT treatment group(L-HX; 130 mmol/L glucose +7.5 mg/L HXMMT), and high-dose HXMMT treatment group(H-HX; 130 mmol/L glucose +75 mg/L HXMMT), with a 3-day intervention period until 6 dpf. The area and length of eyes, and body length of zebrafish were observed by stereomicroscopy, retinal morphology was observed by hematoxylin-eosin staining(HE), and retinal vessel diameter was observed under fluorescence microscope. Differentially expressed genes(DEGs)were identified by RNA-sequencing(RNA-seq)technology to further elucidate the molecular mechanism of HXMMT in improving DR in zebrafish, and the sequencing accuracy was validated through quantitative real-time polymerase chain reaction(qRT-PCR).RESULTS: HE staining demonstrated that the intervention with HXMMT significantly improved the disordered cell arrangement, widened gaps, and thickened inner nuclear layer(INL)in ganglion cell layer GCL); retinal vascular diameter quantification revealed that the retinal vessel diameter of the MG significantly increased compared with the CG, and it was significantly changed after the intervention of HXMMT, with significant efficacy in the H-HX(P<0.05); transcriptomics profiling identified 1 470 reversed DEGs, predominantly enriched in the AMPK signaling pathway, FoxO signaling pathway, retinal developmental processes, and tight junction regulation. Technical validation confirmed strong correlation between qRT-PCR and RNA-seq data(R2=0.8571, P<0.05).CONCLUSION: HXMMT may improve retinal vascular microcirculation disorders in DR by regulating core targets including vsx1, pde6c, arr3a, plk1, fbp1b, foxo1a, pcna, and cdk1, as well as synergistically modulating processes such as retinal development in camera-type eyes, visual perception, microtubule cytoskeletal organization, tight junctions, and the AMPK signaling pathway, Foxo signaling pathway.

Pristimerin, which is one of the compounds present in Celastraceae and Hippocrateaceae, has antitumor effects. However, its mechanism of action in esophageal squamous cell carcinoma (ESCC) remains unclear. This study aims to investigate the efficacy and mechanism of pristimerin on ESCC in vitro and in vivo. The inhibitory effect of pristimerin on cell growth was assessed using trypan blue exclusion and colony formation assays. Cell apoptosis was evaluated by flow cytometry. Gene and protein expressions were analyzed through quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blotting, and immunohistochemistry. RNA sequencing (RNA-Seq) was employed to identify significantly differentially expressed genes (DEGs). Cell transfection and RNA interference assays were utilized to examine the role of key proteins in pristimerin?s effect. Xenograft models were established to evaluate the antitumor efficiency of pristimerin in vivo. Pristimerin inhibited cell growth and induced apoptosis in ESCC cells. Upregulation of Noxa was crucial for pristimerin-induced apoptosis. Pristimerin activated the Forkhead box O3a (FoxO3a) signaling pathway and triggered FoxO3a recruitment to the Noxa promoter, leading to Noxa transcription. Blocking FoxO3a reversed pristimerin-induced Noxa upregulation and cell apoptosis. Pristimerin treatment suppressed xenograft tumors in nude mice, but these effects were largely negated in Noxa-KO tumors. Furthermore, the chemosensitization effects of pristimerin in vitro and in vivo were mediated by Noxa. This study demonstrates that pristimerin exerts an antitumor effect on ESCC by inducing AKT/FoxO3a-mediated Noxa upregulation. These findings suggest that pristimerin may serve as a potent anticancer agent for ESCC treatment.
Forkhead Box Protein O3/genetics* ; Humans ; Apoptosis/drug effects* ; Esophageal Squamous Cell Carcinoma/physiopathology* ; Esophageal Neoplasms/physiopathology* ; Pentacyclic Triterpenes ; Animals ; Cell Line, Tumor ; Proto-Oncogene Proteins c-bcl-2/genetics* ; Mice ; Signal Transduction/drug effects* ; Mice, Nude ; Cell Proliferation/drug effects* ; Triterpenes/pharmacology* ; Xenograft Model Antitumor Assays ; Mice, Inbred BALB C ; Male ; Gene Expression Regulation, Neoplastic/drug effects*

Objective Building a TCM health management and clinical research integration platform for knee osteoarthritis,promoting the integration of traditional Chinese medicine with internet and mobile health technology,driving the digitalization of TCM-enabled knee osteoarthritis health management strategies,and providing a convenient digital platform for the efficient conduct of TCM clinical research on knee osteoarthritis.Methods Using a combination of literature review and market research,a preliminary framework for the platform's functions and services was established.Then,17 patients with knee osteoarthritis were interviewed through a convenient sampling method to identify their needs and preferences,and design the platform's functional modules and service flow.In the development and testing phase,a multidisciplinary team consisting of professional development engineers,data scientists,and clinical experts conducted multiple rounds of discussions to ensure the scientificity,feasibility,and convenience of the platform's function and service development.Results The integrated platform of traditional Chinese medicine health management and clinical research for knee osteoarthritis is built.The traditional Chinese medicine health management module includes features of health education,evaluation and exercise training,etc.The scientific research module includes patient self-report outcome data collection and remote quality control,etc.,which can provide the whole process service integrating health management and remote intelligent clinical trial.Conclusion Building an integrated platform of TCM health management and clinical research will provide more convenient and effective health management solutions for patients with knee osteoarthritis.

Objective:To investigate key genes and cellular communication involved in spinal cord injury (SCI) repair using transcriptome sequencing and single-cell sequencing analysis.Methods:Two transcriptome sequencing datasets related to SCI and one single-cell sequencing dataset were obtained from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) in the two transcriptome sequencing datasets were analyzed using the limma package in R, followed by cross-analysis to identify common DEGs. Functional enrichment analysis of DEGs was performed using the DAVID database for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Hub genes in the protein-protein interaction (PPI) network were identified using the cytoHubba plugin in Cytoscape. Single-cell sequencing data were analyzed using the Seurat package in R. Principal Component Analysis (PCA) was applied for dimensionality reduction, and t-distributed Stochastic Neighbor Embedding (t-SNE) was used for visualization. Cellular communication between different cell groups was analyzed using the CellChat package.Results:Differential analysis of transcriptome sequencing data identified 98 common DEGs. GO and KEGG enrichment analysis highlighted biological processes such as wound healing, macrophage activation, and immune response-related cell activation. The PPI network predicted 10 hub genes: ITGAM, TGFB1, CCL2, ICAM1, CD44, FN1, TIMP1, TLR2, ITGB2, and LGALS3. Single-cell sequencing analysis identified six distinct cell populations, including astrocytes, neurons, and fibroblasts. Cellular communication analysis revealed key signaling pathways between cell subpopulations, including SPP1- (ITGAV+ITGB1), SPP1- (ITGA5+ITGB1), TNF-TNFRSF1A transmission pathway, etc.Conclusions:During SCI repair, astrocytes receive signals from fibroblasts, monocytes, and neurons via the SPP1 pathway, while astrocytes and neurons receive signals from monocytes through the TNF pathway. These findings provide critical insights into the molecular mechanisms underlying SCI repair and offer a foundation for future research.

Objective:To analyze the clinical phenotypes and genetic features of a child with developmental and epileptic encephalopathy due to compound heterozygous variants in the ALG14 gene, and to improve clinicians′ understanding of the ALG14 gene and its associated disorders. Methods:Clinical data of a child with developmental and epileptic encephalopathy caused by ALG14 gene variants, who visited Linyi People′s Hospital on September 9, 2019, were collected and followed up. Whole exome sequencing (WES) and Sanger sequencing were applied to genetically detect the child and her parents, SWISS-MODEL software was used to perform 3D modeling of proteins for the screened ALG14 gene variants, and the literature was reviewed to summarize the clinical phenotypes and genetic characteristics of patients associated with ALG14 gene. Results:The proband, a 17-year-old female, presented with focal seizures and generalized tonic-clonic seizures, generalized developmental delay, cranial magnetic resonance imaging showing no significant abnormalities, and video-electroencephalogram showing widespread low-amplitude fast rhythms. The WES results showed that the affected child carried compound heterozygous variants c.243A>G (p.Ile81Met) and c.103delG(p.Glu35SerfsTer7) (NM_144988) in the ALG14 gene, and Sanger sequencing validation showed that they were derived from her father and mother with normal clinical phenotype, respectively, and the above loci had not been reported domestically or internationally. 3D modeling revealed that the p.Ile81Met variant resulted in a change in the coding amino acid at position 81, and alteration of the stability and hydrophobicity of the protein, which may affect the protein function; the p.Glu35SerfsTer7 variant resulted in early truncation of the peptide chain, loss of a large number of amino acid fragments, and significant alteration of the protein structure. According to the American College of Medical Genetics and Genomics and the Association for Molecular Pathology, p.Ile81Met was a variant of undetermined clinical significance (PM2+PM3+PP3), and p.Glu35SerfsTer7 was a likely pathogenic variant (PVS1+PM2+PP3). A total of 5 papers were retrieved reporting 13 cases related to ALG14 gene variants (including the present case), 4 cases with pure heterozygous variants, 9 with compound heterozygous variants, involving 11 variant types, all inherited from the parents, of which 9 being missense variants, 1 being shifted code variant, and 1 being intronic deletion variant. The main clinical phenotypes included epileptic seizure (11/13), hypotonia (10/13), craniofacial malformations (8/13), growth disorders (3/13), and behavioral abnormalities (2/13). Conclusions:This article reports the first case of ALG14 gene related developmental and epileptic encephalopathy in a child at home and abroad, characterized by focal seizures, generalized tonic-clonic seizures, and global developmental delay. The compound heterozygous variants c.243A>G (p.Ile81Met) and c.103delG(p.Glu35SerfsTer7) may be the genetic cause of this child, enriching the clinical phenotype and variation spectrum related to this gene.