ObjectiveTo investigate the effects of zinc finger E-box binding homeobox 1 （ZEB1） on the proliferation， migration， and invasion of lung adenocarcinoma H322 cells， as well as its underlying molecular mechanisms. MethodsThe gene expression characteristics of the transcription factor ZEB1 in lung adenocarcinoma were analyzed using data from the GEO and TCGA public databases. RT-qPCR and Western blot were employed to measure mRNA and protein expression levels of ZEB1 in lung adenocarcinoma cell lines （H322， A549， 95-D） and normal human bronchial epithelial cells （BEAS-2B）. Lentiviral transduction was utilized to establish stable ZEB1-overexpressing （Oe-ZEB1） and vector control （Oe-NC） H322 cell lines. Cell proliferation was assessed using CCK-8， colony formation， and EdU assays， while apoptosis was evaluated by Hoechst33258/PI double staining. Wound healing and Transwell assays were performed to examine cell migration and invasion capabilities. Cell cycle distribution was determined by flow cytometry， and Western blot was used to analyze protein expression changes in relevant signaling pathways. ResultsThe findings from GEO and TCGA indicated that ZEB1 expression in lung adenocarcinoma varied with tumor malignancy grade. RT-qPCR and Western blot analyses revealed significantly higher ZEB1 expression in lung adenocarcinoma cell lines compared to BEAS-2B cells （P0.05）. Results from the CCK-8， colony formation， EdU， wound healing， and Transwell assays demonstrated that， compared with the un-transfected control （Control） group， Oe-ZEB1 H322 cells exhibited enhanced proliferation， migration， and invasion capabilities （P0.05）. Hoechst33258/PI double staining and flow cytometry analyses showed that， relative to the Control group， apoptosis was reduced in Oe-ZEB1 H322 cells （P0.05）. Additionally， a decreased proportion of cells in the G₁ phase and an increased proportion in the S phase were observed in Oe-ZEB1 cells， indicating accelerated cell cycle progression. Western blot analysis further revealed that， compared with the Control group， Oe-ZEB1 H322 cells exhibited upregulated expression of N-cadherin， mutant p53 （mutp53）， and Cyclin D1 （P0.05）， while expression levels of E-cadherin， murine double minute 2 （MDM2）， and p21 were downregulated （P0.05）. ConclusionOverexpression of ZEB1 promotes the proliferation， migration， and invasion of lung adenocarcinoma H322 cells and may facilitate cell cycle progression by modulating the MDM2/mutp53/p21 signaling pathway， thereby promoting the transition of cells from the G₀/G₁ phase to the S phase.

BACKGROUND:Mitophagy is closely associated with the development of idiopathic pulmonary fibrosis,but its mechanism remains unclear.OBJECTIVE:To investigate the biological mechanism of mitophagy in idiopathic pulmonary fibrosis and provide ideas for the risk prediction of idiopathic pulmonary fibrosis and subtype differentiation.METHODS:The mitophagy-related genes in idiopathic pulmonary fibrosis were obtained through GEO and Reactome Pathway databases.The mitophagy-related characteristic genes in idiopathic pulmonary fibrosis were screened based on intergroup differences and random forest model.GO functional enrichment analysis and KEGG,Reactome with WIKI pathway enrichment analyses were performed by g:Profiler database.Mitophagy subtypes in idiopathic pulmonary fibrosis were distinguished by consensus clustering method and immune infiltration analysis was performed.The mitophagy-related key gene was screened.Finally,the predictive value of mitophagy-related key gene for the risk of idiopathic pulmonary fibrosis was quantified by alignment diagram and the correlation between mitophagy-related key gene and clinical characteristics of idiopathic pulmonary fibrosis was explored.RESULTS AND CONCLUSION:(1)A total of 13 genes related to mitophagy in idiopathic pulmonary fibrosis were identified and 5 characteristic genes were screened,containing PINK1,RPS27A,SRC,HIF1A,and CDH6.(2)GO analysis was mainly involved in ubiquitin protein ligase binding,and cellular response to hypoxia.Pathway enrichment analysis was mainly involved in PINK1-PRKN mediated mitophagy,NOTCH signaling pathway,signaling by EGFR and angiogenesis.(3)HIF1A had significant expression differences between subtypes,which might serve as a key gene for the differentiation of mitophagy subtypes of idiopathic pulmonary fibrosis.(4)Immune infiltration analysis suggested that myeloid-derived suppressor cell,neutrophil and type 1 T helper cell might have infiltration differences between subtypes,while HIF1A was positively correlated with multiple immune cells.(5)Alignment diagram suggested that the risk of idiopathic pulmonary fibrosis might be predicted by the expression level of HIF1A.(6)Clinical characteristics analysis indicated patients with high expression of HIF1A might have poorer lung function and more severe fibrosis.It is concluded that PINK1,RPS27A,SRC,HIF1A,and CDH6 may influence the development of idiopathic pulmonary fibrosis through mitophagy,in which HIF1A may serve as a key gene for risk prediction with clinical subtype differentiation and HIF1A is strongly associated with the lung function of patients.

Objective To investigate the establishment and evaluation of an idiopathic pulmonary fibrosis(IPF)mouse model induced by the intratracheal infusion of bleomycin(BLM)of different concentrations.Methods Male C57BL/6J mice were randomly divided into a control group,Model-L group(1.5 mg/kg,BLM),Model-M group(2.5 mg/kg,BLM),and Model-H group(3.5 mg/kg,BLM).An IPF mouse model was constructed by one-time intratracheal infusion of BLM.The general status,body mass,survival rate,and lung coefficient of mice in different groups were compared.Pathological changes in lung tissue,the hydroxyproline content,fibrosis markers and inflammatory factor levels were observed.Results Compared with the control group,the survival rate decreased and body weight showed a downward trend in the low-,medium-,and high-dose model groups,with significant increases in lung coefficients.Inflammatory infiltration(P＜0.01)and collagen deposition(P＜0.0001)were observed in the lung tissues of all model groups.Hydroxyproline levels in lung tissue and serum were significantly elevated(P＜0.05).The mRNA levels of fibrosis markers α-Sma,Fn1,and Col1a1 were upregulated(P＜0.001),with significant increases in corresponding protein expression(P＜0.05).The mRNA expression of the inflammatory factor Tgfb1 also increased(P＜0.0001).Conclusion 1.5,2.5 and 3.5 mg/kg BLM can induce an IPF model in C57BL/6J mice.Based on the results observed for survival rate,body mass,lung coefficient changes,lung tissue gross and pathological changes,and fibrosis-related biomarkers,2.5 mg/kg BLM is the optimal concentration for inducing an IPF mouse model.

OBJECTIVE: This study investigates the efficacy and mechanisms of Qingjie Fuzheng Granules (QFG) in inhibiting colitis-associated colorectal cancer (CAC) development via RNA sequencing (RNA-seq) and 16S ribosomal RNA (rRNA) correlation analysis. METHODS: CAC was induced in BALB/c mice using azoxymethane (AOM) and dextran sulfate sodium (DSS), and QFG was administered orally to the treatment group. The effects of QFG on CAC were evaluated using disease index, histology, and serum T-cell ratios. RNA-seq and 16S rRNA analysis assessed the transcriptome and microbiome change. Key pharmacodynamic pathways were identified by integrating these data and confirmed via Western blotting and immunofluorescence. The link between microbiota and CAC-related markers was explored using linear discriminant analysis effect size and Spearman correlation analysis. RESULTS: Long-term treatment with QFG prevented AOM/DSS-induced CAC formation, reduced levels of interleukin (IL)-1β, tumor necrosis factor-alpha (TNF-α), IL-6, and interferon γ (IFN-γ), and increased CD3⁺ and CD4⁺/CD8⁺ T cells ratio, without causing hepatic or renal toxicity. A 16S rRNA analysis revealed that QFG rebalanced the Firmicutes/Bacteroidetes ratio and mitigated AOM/DSS-induced microbiota disturbances. Transcriptomics and Western blotting analysis identified the nucleotide-binding oligomerization domain-containing protein 2 (NOD2)/nuclear factor kappa-B (NF-κB) pathway as key for QFG's treatment against CAC. Furthermore, QFG decreased the abundance of Bacilli, Bacillales, Staphylococcaceae, Staphylococcus, Lactobacillales, Aerococcus, Alloprevotella, and Akkermansia, while increasing Clostridiales, Lachnospiraceae, Lachnospiraceae_NK4A136_group, Ruminococcaceae, and Muribaculaceae, which were highly correlated with CAC-related markers or NOD2/NF-κB pathway. CONCLUSION By mapping the relationships between CAC, immune responses, microbiota, and key pathways, this study clarifies the mechanism of QFG in inhibiting CAC, highlighting its potential for clinical use as preventive therapy.

Objective To investigate the establishment and evaluation of an idiopathic pulmonary fibrosis(IPF)mouse model induced by the intratracheal infusion of bleomycin(BLM)of different concentrations.Methods Male C57BL/6J mice were randomly divided into a control group,Model-L group(1.5 mg/kg,BLM),Model-M group(2.5 mg/kg,BLM),and Model-H group(3.5 mg/kg,BLM).An IPF mouse model was constructed by one-time intratracheal infusion of BLM.The general status,body mass,survival rate,and lung coefficient of mice in different groups were compared.Pathological changes in lung tissue,the hydroxyproline content,fibrosis markers and inflammatory factor levels were observed.Results Compared with the control group,the survival rate decreased and body weight showed a downward trend in the low-,medium-,and high-dose model groups,with significant increases in lung coefficients.Inflammatory infiltration(P＜0.01)and collagen deposition(P＜0.0001)were observed in the lung tissues of all model groups.Hydroxyproline levels in lung tissue and serum were significantly elevated(P＜0.05).The mRNA levels of fibrosis markers α-Sma,Fn1,and Col1a1 were upregulated(P＜0.001),with significant increases in corresponding protein expression(P＜0.05).The mRNA expression of the inflammatory factor Tgfb1 also increased(P＜0.0001).Conclusion 1.5,2.5 and 3.5 mg/kg BLM can induce an IPF model in C57BL/6J mice.Based on the results observed for survival rate,body mass,lung coefficient changes,lung tissue gross and pathological changes,and fibrosis-related biomarkers,2.5 mg/kg BLM is the optimal concentration for inducing an IPF mouse model.

BACKGROUND:Mitophagy is closely associated with the development of idiopathic pulmonary fibrosis,but its mechanism remains unclear.OBJECTIVE:To investigate the biological mechanism of mitophagy in idiopathic pulmonary fibrosis and provide ideas for the risk prediction of idiopathic pulmonary fibrosis and subtype differentiation.METHODS:The mitophagy-related genes in idiopathic pulmonary fibrosis were obtained through GEO and Reactome Pathway databases.The mitophagy-related characteristic genes in idiopathic pulmonary fibrosis were screened based on intergroup differences and random forest model.GO functional enrichment analysis and KEGG,Reactome with WIKI pathway enrichment analyses were performed by g:Profiler database.Mitophagy subtypes in idiopathic pulmonary fibrosis were distinguished by consensus clustering method and immune infiltration analysis was performed.The mitophagy-related key gene was screened.Finally,the predictive value of mitophagy-related key gene for the risk of idiopathic pulmonary fibrosis was quantified by alignment diagram and the correlation between mitophagy-related key gene and clinical characteristics of idiopathic pulmonary fibrosis was explored.RESULTS AND CONCLUSION:(1)A total of 13 genes related to mitophagy in idiopathic pulmonary fibrosis were identified and 5 characteristic genes were screened,containing PINK1,RPS27A,SRC,HIF1A,and CDH6.(2)GO analysis was mainly involved in ubiquitin protein ligase binding,and cellular response to hypoxia.Pathway enrichment analysis was mainly involved in PINK1-PRKN mediated mitophagy,NOTCH signaling pathway,signaling by EGFR and angiogenesis.(3)HIF1A had significant expression differences between subtypes,which might serve as a key gene for the differentiation of mitophagy subtypes of idiopathic pulmonary fibrosis.(4)Immune infiltration analysis suggested that myeloid-derived suppressor cell,neutrophil and type 1 T helper cell might have infiltration differences between subtypes,while HIF1A was positively correlated with multiple immune cells.(5)Alignment diagram suggested that the risk of idiopathic pulmonary fibrosis might be predicted by the expression level of HIF1A.(6)Clinical characteristics analysis indicated patients with high expression of HIF1A might have poorer lung function and more severe fibrosis.It is concluded that PINK1,RPS27A,SRC,HIF1A,and CDH6 may influence the development of idiopathic pulmonary fibrosis through mitophagy,in which HIF1A may serve as a key gene for risk prediction with clinical subtype differentiation and HIF1A is strongly associated with the lung function of patients.

Fetal monitoring is an essential component of the prenatal examination. With electronic fetal heart monitoring, clinicians can effectively monitor the intrauterine situation of the fetus, promptly detect fetal distress, and intervene early to reduce the occurrence of adverse outcomes in newborns. In recent years, the leaps in internet technology have enabled the widespread utilization of remote electronic fetal heart monitoring based on ultrasound technology. This paper reviews the application, effectiveness, and safety of remote fetal heart monitoring, and the satisfaction level of healthcare professionals with this technology in recent years and compares it with traditional fetal heart monitoring, aiming to provide reference and insights for clinical applications of remote fetal heart monitoring.

Objective The goals of this study were to identify early warning markers of severe dengue based on bioinformatics com-bined with machine learning,and explore the evaluation system of the risk of occurrence of severe dengue.Methods Based on the Gene Expression Omnibus database,the differentially expressed genes between dengue and severe dengue were analyzed,and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted.Early warning genes of severe dengue were screened using a random forest model,and the accuracy of the genes was verified using receiver operating characteristic(ROC)curves.Finally,nomograms were constructed to quantify the warning genes and predict the risk of progression from dengue to severe dengue based on the expression level of these genes.Results A total of 817 differentially expressed genes were identified,along with the associated biolo-gical processes that may be closely related to the occurrence and development of severe dengue,namely,antimicrobial humoral response,humoral immune response,serine hydrolase activity,and arachidonic acid metabolism.Based on this analysis,five early warning genes were isolated:AZU1,PDCD4,COL4A3BP,TRPM4,and ATP4A.Among these,ATP4A,COL4A3BP,and TRPM4 showed low expression levels,whereas AZU1and PDCD4were highly expressed.The ROC curves indicated that these genes were accurate pre-dictors of severe dengue.The nomograms indicated good predictive accuracy,clinical benefit rate,and clinical effectiveness of the model.Conclusion Measuring the expression levels of five warning genes(AZU1,PDCD4,COL4A3BP,TRPM4,and ATP4A)may help to evaluate the risk of severe dengue.

Objective:To monitor intra-fractional set-up errors in tumor radiotherapy using a real-time intelligent capture system for precision displacement.Methods:A simulated radiotherapy environment was created in both the laboratory and the treatment room. A three-axis ( xyz) displacement platform (LD60-LM) and dial gauges were used as displacement measurement tools. Moreover, a real-time intelligent capture system for precision displacement was developed for displacement monitoring. With 23 patients treated with radiotherapy enrolled in this study, the above system was employed to monitor their intra-fractional set-up errors in fractionated radiotherapy. Descriptive analyses were conducted on the deviations between the data captured by cameras and the actual displacement, obtaining the mean values and standard deviation. Results:The monitoring calibration data from the laboratory revealed displacement differences of ≤ 0.5 mm within 20 mm and a maximum displacement difference of 1.47 mm for 50 mm. In contrast, the calibration result from the treatment room exhibited deviations of ± 0.2 mm on the y- z axes, as displayed by both the left and right cameras, and ± 0.31 mm on the x- z axes, as displayed by the middle camera. During 37 radiotherapy sessions in 23 patients, the monitoring result from the middle camera revealed five deviations exceeding the threshold of 5 mm, with the maximum deviation duration and displacement of 57.2 s and 9.24 mm, respectively. Conclusions:The real-time intelligent capture system for precision displacement based on machine vision can achieve real-time monitoring of set-up errors during tumor radiotherapy. Nevertheless, further improvements and service testing are necessary for this system.