ObjectiveTo explore the role of the histone deacetylase Sirtuin-1 （SIRT1）/p53 signaling pathway in promoting apoptosis of non-small cell lung cancer cells （NSCLC） induced by the co-stimulatory molecule B7 homolog 3 （B7-H3）. MethodsThe GEPIA 2 platform was used for survival analysis of NSCLC patients based on B7⁃H3 gene expression levels. The Gene Enrichment Analysis （GSEA） method was used to analyze the enrichment characteristics of B7⁃H3 molecules in the gene set of cell apoptosis. In the non-small cell lung cancer A549 cell line， B7⁃H3 was knocked down， and the protein expression levels of SIRT1 and p53 were detected by Western blot. B7⁃H3 was overexpressed in A549 cells and the apoptosis rate was analyzed by flow cytometry after Annexin V/PI double staining. Overexpression of B7⁃H3 and knockdown of SIRT1 were performed in A549 cell line. The expression levels of p53 and apoptosis-related proteins B-cell lymphoma/leukemia-2 （Bcl-2） and Bcl-2-associated X protein （Bax） were detected respectively by Western blot. Cell apoptosis rate was analyzed by flow cytometry after Annexin V/PI double staining. ResultsThe overall survival of the B7-H3 high-expression group was significantly lower than that of the low-expression group （P<0.01）. B7-H3 was significantly enriched in the cell apoptosis signaling pathway and the p53 signaling pathway （P<0.05）. Compared with the control group， the expression of SIRT1 was significantly downregulated， and p53 was significantly upregulated in the B7⁃H3 knockdown group （both P<0.001）. Overexpression of B7-H3 significantly up-regulated SIRT1 protein expression （P<0.05）， down-regulated p53 expression （P<0.01）， and markedly increased the Bcl-2/Bax ratio of apoptosis-related proteins （P<0.001）. The results of Annexin V/PI double staining showed that the apoptosis rate of A549 cells with overexpressed B7⁃H3 decreased （the apoptosis rate of the control group was 26.72%±4.13%， while that of the B7⁃H3 overexpression group was 13.87%±0.82%； P<0.01）. In B7-H3-overexpressing cell lines， SIRT1 knockdown significantly reversed apoptosis （P<0.05）， up-regulated p53 protein expression （P<0.001）， and markedly reduced the Bcl-2/Bax ratio （P<0.001）. ConclusionB7-H3 molecule inhibits the apoptosis of non-small cell lung cancer cells via the SIRT1/p53 signaling pathway.

BACKGROUND: This study aimed to investigate programmed death-ligand 1 tumor proportion score in predicting the safety and efficacy of PD-1/PD-L1 antibody-based therapy in treating patients with advanced non-small cell lung cancer (NSCLC) in a real-world setting. METHODS: This retrospective, multicenter, observational study enrolled adult patients who received PD-1/PD-L1 antibody-based therapy in China and met the following criteria: (1) had pathologically confirmed, unresectable stage III-IV NSCLC; (2) had a baseline PD-L1 tumor proportion score (TPS); and (3) had confirmed efficacy evaluation results after PD-1/PD-L1 treatment. Logistic regression, Kaplan-Meier analysis, and Cox regression were used to assess the progression-free survival (PFS), overall survival (OS), and immune-related adverse events (irAEs) as appropriate. RESULTS: A total of 409 patients, 65.0% ( n = 266) with a positive PD-L1 TPS (≥1%) and 32.8% ( n = 134) with PD-L1 TPS ≥50%, were included in this study. Cox regression confirmed that patients with a PD-L1 TPS ≥1% had significantly improved PFS (hazard ratio [HR] 0.747, 95% confidence interval [CI] 0.573-0.975, P = 0.032). A total of 160 (39.1%) patients experienced 206 irAEs, and 27 (6.6%) patients experienced 31 grade 3-5 irAEs. The organs most frequently associated with irAEs were the skin (52/409, 12.7%), thyroid (40/409, 9.8%), and lung (34/409, 8.3%). Multivariate logistic regression revealed that a PD-L1 TPS ≥1% (odds ratio [OR] 1.713, 95% CI 1.054-2.784, P = 0.030) was an independent risk factor for irAEs. Other risk factors for irAEs included pretreatment absolute lymphocyte count >2.5 × 10 9 /L (OR 3.772, 95% CI 1.377-10.329, P = 0.010) and pretreatment absolute eosinophil count >0.2 × 10 9 /L (OR 2.006, 95% CI 1.219-3.302, P = 0.006). Moreover, patients who developed irAEs demonstrated improved PFS (13.7 months vs. 8.4 months, P <0.001) and OS (28.0 months vs. 18.0 months, P = 0.007) compared with patients without irAEs. CONCLUSIONS A positive PD-L1 TPS (≥1%) was associated with improved PFS and an increased risk of irAEs in a real-world setting. The onset of irAEs was associated with improved PFS and OS in patients with advanced NSCLC receiving PD-1/PD-L1-based therapy.
Humans ; Carcinoma, Non-Small-Cell Lung/metabolism* ; Male ; Female ; Retrospective Studies ; Middle Aged ; Lung Neoplasms/metabolism* ; Aged ; B7-H1 Antigen/metabolism* ; Programmed Cell Death 1 Receptor/metabolism* ; Adult ; Aged, 80 and over ; Immune Checkpoint Inhibitors/therapeutic use*

Objective:To evaluate the role of interferon gene stimulator/long-chain ester acyl-CoA synthetase 4 (STING/ACSL4) signaling pathway in alleviation of oxygen-glucose deprivation and restoration (OGD/R)-induced ferroptosis in renal tubular epithelial cells.Methods:The close juxial tubule epithelial cells of human renal cortex were selected and divided into 9 groups ( n=78 each) using a random number table method: control group (C group ), OGD/R group, OGD/R + 25 μmol/L ciprofol group (HC25 group), OGD/R + 50 μmol/L ciprofol group (HC50 group), OGD/R + 100 μmol/L ciprofol group (HC100 group), virus control (NC) group, OGD/R + NC group, OGD/R + ciprofol + NC group (OGD/R+ Cip+ NC group), and OGD/R + ciprofol + STING overexpression lentivirus group (OGD/R+ Cip+ OE-STING group). The OGD/R model was developed by subjecting the cells to O 2-glucose deprivation (OGD) for 4 h followed by restoration of O 2-glucose supply for 20 h. Ciprofol at a final concentration of 25, 50 and 100 μmol/L was added to the medium during OGD/R in HC25, HC50, and HC100 groups, respectively. The cells were subjected to conventional culture after infection with the control virus of the STING overexpression lentivirus in NC group. The OGD/R model was developed after the cells were infected with control virus in OGD/R+ NC group. In OGD/R+ Cip+ NC group and OGD/R+ Cip+ OE-STING group, the cells were infected with control virus and STING overexpression lentivirus, respectively, and ciprofol 50 μmol/L was added to the medium during OGD/R. Cell damage parameters included the cell viability and activity of lactic dehydrogenase (LDH) in supernatant. The oxidative stress parameters included the activity of reactive oxygen species (ROS) and contents of malondialdehyde (MDA) and glutathione (GSH). Mitochondrial damage parameters included the mitochondrial area and branch length, content of mitochondrial 8-hydroxydeoxyguanosine (8-OHdG) in mitochondrial DNA (mtDNA), and DNA expression of nicotinamide adenine dinucleotide dehydrogenase 1 and 2 (mtND1, mtND2) and cytochrome oxidase (COX-1). The ferroptosis parameters included Fe 2+ content and expression of STING, ACSL4, nuclear factor-κB (NF-κB), cyclic GMP-AMP synthase (cGAS), and glutathione peroxidase 4 (GPX4) protein and mRNA. Results:Compared with group C, the activity of LDH in the supernatant was significantly increased, the cell viability was decreased, the ROS activity, MDA content, and Fe 2+ content were increased, the GSH content was decreased, the expression of ACSL4, cGAS, STING, NF-κB protein and mRNA was up-regulated, the expression of GPX4 protein and mRNA was down-regulated, the content of 8-OHdG in mtDNA was increased, the DNA expression of cytoplasmic mtND1, mtND2 and COX-1 was up-regulated, and the mitochondrial area and branch length were increased in group OGD/R ( P<0.05). Compared with OGD/R group, the cell viability and GSH content were significantly increased, the MDA content and Fe 2+ content were decreased, the expression of ACSL4, cGAS, STING, NF-κB protein and mRNA was down-regulated, the expression of GPX4 protein and mRNA was up-regulated, the content of 8-OHdG in mtDNA was decreased, and the DNA expression of cytoplasmic mtND1, mtND2 and COX-1 was up-regulated in HC50 group ( P<0.05). Compared with OGD/R+ Cip+ NC group, the cell viability was significantly decreased, the ROS activity was increased, the expression of ACSL4, cGAS and NF-κB protein and mRNA was up-regulated, the expression of GPX4 protein and mRNA was down-regulated, and the DNA expression of cytoplasmic mtND1, mtND2 and COX-1 was up-regulated in OGD/R+ Cip+ OE-STING group ( P<0.05). Conclusions:Ciprofol may exert cytoprotective effects by alleviating ferroptosis during OGD/R in renal tubular epithelial cells by inhibiting STING/ACSL4 signaling pathway.

Objective:To develop an artificial intelligence system using fully convolutional neural networks（FCN）to assist echocardiographers in the quantitative assessment of mitral regurgitation（MR）severity.Methods:From August 2021 to June 2024，echocardiographic images of 441 patients with MR were prospectively collected from Renmin Hospital of Wuhan University and the Central Hospital of Wuhan. After screening，a total of 269 patients（4 917 frames）were included in the study. Of these，3 644 frames（128 patients）of apical four-chamber color Doppler MR flow convergence images from Renmin Hospital of Wuhan University were selected as the training/validation set，while images from 121 patients（813 frames）were used as the internal test set. Additionally，images from 20 patients（460 frames）from the Central Hospital of Wuhan were selected as the external test set. The FCN algorithm was employed to capture features and segment the MR color region on the left atrial side，simultaneously outputting the regurgitant radius（r）for the calculation of the effective regurgitant orifice area and regurgitant volume. The severity of MR was then classified according to the 2017 guidelines of the American Society of Echocardiography. The segmentation and classification performance of the model was evaluated，and the measurement results of the AI system was compared with that of both senior and junior physicians.Results:In the internal test set，the accuracy of r identification for cases classified as Grade Ⅰ to Ⅳ was 0.48，0.81，0.86，and 0.87，respectively. In the external test set，the accuracy of r identification for cases classified as Grade Ⅰ to Ⅳ was 0.60，0.77，0.64，and 0.77，respectively. The average accuracy of MR classification in the internal and external test sets was 0.91 and 0.88，respectively.Conclusions:The FCN model is capable of segmenting the left atrial side regurgitant areas in apical four-chamber heart color Doppler images，aiding physicians in obtaining quantitative assessment parameters for MR，and assisting junior physicians in accurately assessing the severity of MR.

Objective To analyze and preliminarily verify key genes and pathways in the transcriptome of peripheral blood of lung adenocarcinoma patients with metastasis bone pain(MBP),and to explore its underlying mechanism.Methods Nine lung adenocarcinoma patients with bone metastasis treated in the First Affiliated Hospital of Sun Yat-sen University from May 2020 to May 2021 were selected for retrospective analysis,including 4 patients with typical MBP clinical manifestations and visual analogue scale(VAS)≥4(MBP group)and 5 patients without suffering any pain(control group).Peripheral blood mRNA sequencing was performed to identify differentially expressed genes(DEGs),followed by functional pathways analysis and protein-protein interaction(PPI)network analysis.The most significant modules and hub genes were confirmed and visualized using Cytoscape software.The target miRNAs regulating these hub genes were predicted using Targetscan database,and long non-coding RNA(lncRNA)interacting with these miRNAs were also predicted using lncBase database.The relationships among lncRNA,miRNA and mRNA were visualized to construct a competing endogenous RNA(ceRNA)network through Cytoscape software,and the nodes of this network were verified using quantitative PCR(qPCR).Results A total of 1466 DEGs were identified,including 666 up-regulated genes and 800 down-regulated genes.Chemokine receptor 3(CXCR3),pro-opiomelanocortin(POMC),neuromedin U receptor 1(NMUR1),chemokine ligand 2(CCL2)and endocannabinoid receptor 1(CNR1)were identified as hub genes.The most significant enriched processes and pathways of DEGs included osteoclast differentiation,NOD like receptor signal transduction pathway,type Ⅰinterferon signal pathway,nuclear factor kappa-B(NF-κB)signal pathway,apoptosis/autophagy pathway,chemokine signal pathway,interleukin(IL)-1β pathway.Two ceRNA networks were identified:MALAT1-hsa-miR-124-3p.2-CCL2 and NEAT1-hsa-miR-325-3p-CXCR3.qPCR results showed that the expression levels of CCL2,CXCR3,MALAT1,NEAT1 and hsa-miR-325 were higher in MBP group than these in control group(P<0.05).Conclusions CXCR3,POMC,NMUR1,CCL2 and CNR1 may serve as key genes in the occurrence of MBP and could be important regulatory targets for MBP.The development of MBP in lung adenocarcinoma may be associated with the dysregulation of the networks:MALAT1-hsa-miR-124-3p.2-CCL2 and NEAT1-hsa-miR-325-3p-CXCR3.

Objective To initially explore the possibility of applying the fluorescence micro-optical sectioning tomography(fMOST)high-resolution 3D reconstruction system to the morphological study of the intestinal nervous system and to preliminarily establish a method for studying the morphology of the intestinal nervous system using this system.Methods fMOST high-resolution 3D reconstruction system was used to study the intestinal nervous system of C57BL/6 mice in detail.Based on this method,a new morphological method of the visceral nervous system of small animal models was explored at the single-cell level.Results Compared with the large intestine,the small intestine lacked the typical myenteric plexus(Auerbach),deep mucosal plexus(Henley),and submucosal superficial plexus(Meissner).Conclusion The result of this paper provide a clearer and systematic display of the anatomical structure of the enteric nervous system in C57BL/6 mice,and further clarify the similarities and differences between the enteric nervous system of mice and human,and provide a theoretical basis for its rational application in the study of digestive system diseases.The morphological study of fMOST high-resolution 3D reconstruction system is not limited to the central nervous system,but can be extended to the morphological study of multiple visceral nervous systems.

Diffuse pediatric-type high-grade glioma (pHGG), H3- and isocitrate dehydrogenase (IDH)-wild-type, is a World Health Organization (WHO) grade 4 diffuse glioma with malignant histological features, which typically affects in children and adolescents. This multidisciplinary treatment case involves a 24-year-old young female patient initially diagnosed via biopsy. After concurrent chemoradiotherapy reduced the tumor size, complete resection was achieved, followed by adjuvant therapy with temozolomide and regorafenib. After recurrence, the tumor was resected again, and a second course of radiotherapy was administered, but the disease ultimately progressed rapidly, with an overall survival exceeding 32 months. pHGGs are highly malignant, aggressive, and associated with poor prognosis. Accurate diagnosis and targeted intervention require close collaboration among a multidisciplinary team, which highlights the critical value of multidisciplinary manage-ment in clinical practice.

OBJECTIVE: To elucidate the specific mechanisms by which electroacupuncture (EA) alleviates anxiety and fear behaviors associated with posttraumatic stress disorder (PTSD), focusing on the role of lipocalin-2 (Lcn2). METHODS: The PTSD mouse model was subjected to single prolonged stress and shock (SPS&S), and the animals received 15 min sessions of EA at Shenmen acupoint (HT7). Behavioral tests were used to investigate the effects of EA at HT7 on anxiety and fear. Western blotting and enzyme-linked immunosorbent assay were used to quantify Lcn2 and inflammatory cytokine levels in the prefrontal cortex (PFC). Additionally, the activity of PFC neurons was evaluated by immunofluorescence and in vivo electrophysiology. RESULTS: Mice subjected to SPS&S presented increased anxiety- and fear-like behaviors. Lcn2 expression in the PFC was significantly upregulated following SPS&S, leading to increased expression of the proinflammatory cytokines tumor necrosis factor-α and interleukin-6 and suppression of PFC neuronal activity. However, EA at HT7 inhibited Lcn2 release, reducing neuroinflammation and hypoexcitability in the PFC. Lcn2 overexpression mitigated the effects of EA at HT7, resulting in anxiety- and fear-like behaviors. CONCLUSION EA at HT7 can ameliorate PTSD-associated anxiety and fear, and its mechanism of action appears to involve the inhibition of Lcn2-mediated neural activity and inflammation in the PFC. Please cite this article as: Yang YD, Zhong W, Chen M, Tang QC, Li Y, Yao LL, et al. Electroacupuncture alleviates behaviors associated with posttraumatic stress disorder by modulating lipocalin-2-mediated neuroinflammation and neuronal activity in the prefrontal cortex. J Integr Med. 2025; 23(5):537-547.
Electroacupuncture ; Stress Disorders, Post-Traumatic/metabolism* ; Animals ; Lipocalin-2/metabolism* ; Prefrontal Cortex/physiopathology* ; Male ; Mice ; Neurons/physiology* ; Disease Models, Animal ; Fear ; Behavior, Animal ; Mice, Inbred C57BL ; Neuroinflammatory Diseases/metabolism* ; Anxiety/therapy* ; Acupuncture Points

OBJECTIVE: To investigate the relationship between physical activity and genetic risk and their combined effects on the risk of developing chronic obstructive pulmonary disease. METHODS: This prospective cohort study included 318,085 biobank participants from the UK. Physical activity was assessed using the short form of the International Physical Activity Questionnaire. The participants were stratified into low-, intermediate-, and high-genetic-risk groups based on their polygenic risk scores. Multivariate Cox regression models and multiplicative interaction analyses were used. RESULTS: During a median follow-up period of 13 years, 9,209 participants were diagnosed with chronic obstructive pulmonary disease. For low genetic risk, compared to low physical activity, the hazard ratios ( HRs) for moderate and high physical activity were 0.853 (95% confidence interval [ CI]: 0.748-0.972) and 0.831 (95% CI: 0.727-0.950), respectively. For intermediate genetic risk, the HRs were 0.829 (95% CI: 0.758-0.905) and 0.835 (95% CI: 0.764-0.914), respectively. For participants with high genetic risk, the HRs were 0.809 (95% CI: 0.746-0.877) and 0.818 (95% CI: 0.754-0.888), respectively. A significant interaction was observed between genetic risk and physical activity. CONCLUSION Moderate or high levels of physical activity were associated with a lower risk of developing chronic obstructive pulmonary disease across all genetic risk groups, highlighting the need to tailor activity interventions for genetically susceptible individuals.
Humans ; Pulmonary Disease, Chronic Obstructive/epidemiology* ; Exercise ; Male ; Female ; Middle Aged ; Prospective Studies ; Aged ; Genetic Predisposition to Disease ; Risk Factors ; United Kingdom/epidemiology* ; Incidence ; Adult