Objective To investigate the role and molecular mechanism of neural precursor cell expressed, developmentally down-regulated 9 (NEDD9) in the invasion and migration of colorectal cancer (CRC) cells. Methods Human CRC HCT116 cells were divided into the following groups: HCT116 group (conventional culture); HCT116+U0126 (ERK pathway inhibitor) group (treated with 10 μmol/L U0126 for 24 hours); shNC group (negative control) transfected with shNC lentivirus; shNEDD9 group (NEDD9 knockdown) transfected with shNEDD9 lentivirus; in addition, oeNC group (negative control) transfected with oeNC lentivirus; oeNEDD9 group (NEDD9 overexpression) infected with oeNEDD9 lentivirus; oeNC+U0126 group and oeNEDD9+U0126 group (treated with 10 μmol/L U0126 for 24 hours in addition to the treatment in the oeNC group and oeNEDD9 group). The RT-qPCR technique was used to measure the relative mRNA expression levels of NEDD9, E-cadherin, Vimentin, and N-cadherin; Western blotting was used to measure the relative protein expression levels of NEDD9, E-cadherin, p-ERK and ERK; scratch assay and Transwell assay were used to observe cell migration and invasion abilities. Results Scratch assay and Transwell assay showed that the shNEDD9 group had significantly lower migration and invasion rates than the shNC group, and the HCT116+U0126 group had significantly lower migration and invasion rates than the HCT116 group (t=4.735-7.167,P<0.01). The results of RT-qPCR showed that compared with the shNC group, the shNEDD9 group had a significant increase in the relative mRNA expression level of E-cadherin and significant reductions in the relative mRNA expression levels of Vimentin and N-cadherin (t=3.102-7.121,P<0.05). Western blotting showed that compared with the shNC group, the shNEDD9 group had a significant increase in the relative protein expression level of E-cadherin and a significant reduction in p-ERK/ERK ratio (t=3.054,4.851,P<0.05); compared with the oeNC group, the oeNEDD9 group had a significant increase in p-ERK/ERK ratio, while the oeNC+U0126 group had a significant reduction in p-ERK/ERK ratio (F=21.950,q=6.389,7.853,P<0.01); compared with the oeNC+U0126 group, the oeNEDD9+U0126 group had a significant increase in p-ERK/ERK ratio (q=8.034,P<0.01). Conclusion NEDD9 can promote epithelial-mesenchymal transition of CRC cells by activating the ERK pathway, thereby promoting the invasion and migration of CRC cells.

Objective To investigate the effect of celastrol on the repair of sciatic nerve injury in rats and its mechanism of action. Methods Six-week-old male Sprague-Dawley (SD) rats were divided into groups A-C and groups A1-C1. Groups A and A1 were sham-operation groups; the rats in groups B and B1 were used to establish a model of sciatic nerve injury; the rats in groups C and C1 were given intraperitoneal injection of celastrol at a dose of 1 mg/kg after sciatic nerve injury. Sciatic nerve function index (SFI) was determined for the rats in groups A-C on days 1, 3, 7, 14, 21 and 28 after modeling; sciatic nerve tissue of the injured side was harvested from the rats in groups A1-C1 on day 7, while sciatic nerve tissue of the injured side was harvested from those in groups A-C on day 28, and the wet weight ratio of the gastrocnemius muscle was calculated. The rat Schwann cell line RSC96 was divided into groups E1-E8. The cells in group E1 were not given any treatment; the cells in group E2 were treated with 0.4 μmol/L erastin, and those in groups E3-E8 were treated with 0.4 μmol/L erastin and celastrol at different concentrations of 0.1, 0.2, 0.4, 0.8, 1.6 and 3.2 μmol/L, respectively. In addition, RSC96 cells were divided into groups F1-F3: the cells in group F1 were not given any treatment; the cells in group F2 were treated with 0.4 μmol/L erastin; the cells in group F3 were treated with 0.4 μmol/L erastin and 0.1 μmol/L celastrol. Immunohistochemical staining was used to measure the expression levels of growth-associated protein 43 (GAP-43) and p75 neurotrophin receptor (p75NTR) in sciatic nerve tissue of the rats in groups A-C; CCK-8 assay was used to measure the viability of cells in groups E1-E8; Western blot was used to measure the expression level of glutathione peroxidase 4 (GPX4) in sciatic nerve tissue of the rats in groups A1-C1 and in RSC96 cells from groups F1-F3; a tissue iron assay kit was used to measure the level of Fe²⁺ in sciatic nerve tissue of the rats in groups A1-C1; a malondialdehyde (MDA) assay kit was used to measure the level of MDA in sciatic nerve tissue of the rats in groups A1-C1 and in the cells from groups F1-F3; flow cytometry was used to measure the levels of Fe²⁺ and reactive oxygen species (ROS) in the cells from groups F1-F3. Results The animal experiment showed that compared with group B, group C had significant increases in the wet weight ratio of the gastrocnemius muscle and the expression levels of GAP-43 and p75NTR in sciatic nerve tissue (F=15.17-43.64,q=3.96-6.28,P<0.05); compared with group B on days 21 and 28 after modeling, group C had a significantly higher SFI (t=4.45,5.58,P<0.05); compared with group B1, group C1 had significant reductions in the levels of MDA and Fe²⁺ and a significant increase in the relative protein expression level of GPX4 in sciatic nerve tissue (F=10.59-21.26,q=4.30-7.01,P<0.05). The cell experiment showed that compared with group E2, groups E3-E6 had a significant increase in cell viability (F=331.20,q=10.07-31.54,P<0.05); compared with group F2, group F3 had significant reductions in the levels of MDA, Fe²⁺, and ROS and a significant increase in the relative protein expression level of GPX4 (F=8.49-82.14,q=4.44-13.41,P<0.05). Conclusion Celastrol may promote the repair of sciatic nerve injury in rats by inhibiting ferroptosis in sciatic nerve cells.

Objective To investigate the effect and mechanism of aquaporin 5 (AQP5) on ferroptosis in lens epithelial cells (LECs). Methods Immortalized human LECs (SRA01/04 cells) were divided into empty lentiviral vector control group (group A), stable AQP5 knockdown group (group B), and stable AQP5 knockdown+mitochondria-targeted antioxidant treatment group (group C). Related kits were used to measure mitochondrial reactive oxygen species (mtROS), mitochondrial membrane potential (MMP), and mitochondrial permeability transition pore (mPTP) fluorescence intensity; fluorescent probes were used to measure mitochondrial lipid peroxidation and the level of Fe²⁺; Western blot and immunofluorescence staining were used to measure the protein expression levels of Nrf2, SLC7A11, and GPX4. Results Compared with group A, group B had significant increases in the level of mtROS, the level of mitochondrial lipid peroxides, and the level of mitochondrial Fe²⁺ and significant reductions in MMP, the fluorescence intensity of mPTP and the relative protein expression levels of Nrf2, SLC7A11, and GPX4. Compared with group B, group C had significant reductions in the level of mtROS, the level of mitochondrial lipid peroxides, and the level of mitochondrial Fe²⁺ and significant increases in MMP, the fluorescence intensity of mPTP and the relative protein expression levels of Nrf2, SLC7A11, and GPX4 (F=5.66-46.17,q=2.88-9.32,P<0.05). Conclusion AQP5 deficiency can induce ferroptosis in LECs by inhibiting the Nrf2-SLC7A11-GPX4 axis. Maintaining or upregulating the expression level of AQP5 in LECs may become a potential strategy for the prevention of lens opacity and the treatment of cataract.

Objective To investigate the regulatory effect of DNA methyltransferase 1 (DNMT1) and DNA methyltransferase 3A (DNMT3A) on autophagy of gastric cancer cells. Methods Western blot was used to measure the protein expression levels of DNMT1, DNMT3A, and DNA methyltransferase 3B (DNMT3B) in human gastric mucosal epithelial cell line GES-1 and gastric cancer cell line AGS-EBV. According to experimental requirements, AGS-EBV cells were divided into groups A-D, and human gastric cancer GT38 cells were divided into groups a-d. The cells in groups A/a and groups B/b were transfec-ted with siNC and siDNMT1, respectively, and those in groups C/c and groups D/d were transfected with siNC and siDNMT3A, respectively. Western blot was used to measure the expression levels of DNMT1/DNMT3A and p62 and LC3Ⅱ/Ⅰ ratio in groups A-D and groups a-d; immunofluorescence assay was used to measure the number of LC3 autophagosomes in groups A-D; transmission electron microscopy was used to measure the number of autolysosomes in groups A-D. Results Western blot showed that the relative protein expression levels of DNMT1, DNMT3A, and DNMT3B in AGS-EBV cells were significantly hig-her than those in GES-1 cells (t=2.947-6.695,P<0.05). Compared with groups A and a, groups B and b had a significant reduction in the relative protein expression level of DNMT1 and significant increases in the relative protein expression level of p62 protein and LC3Ⅱ/Ⅰ ratio (t=2.828-126.800,P<0.05). Compared with groups C and c, groups D and d had significant reductions in the relative protein expression levels of DNMT3A and p62 and a significant increase in LC3Ⅱ/Ⅰ (t=3.215-96.110,P<0.05). Immunofluorescence assay and transmission electron microscopy showed that group B had significantly higher numbers of LC3 autophagosomes and autolysosomes than group A, and group D had significantly higher numbers than group C (t=3.200-5.701,P<0.05). Conclusion The expression levels of DNMT1 and DNMT3A in gastric cancer cells are significantly upregulated compared with those in human gastric mucosal epithelial cell line GES-1, suggesting that gastric cancer cells may exhibit a hypermethylation state, and inhibition of DNMT1/DNMT3A expression in gastric cancer cells can enhance the level of autophagy.

Objective To investigate the effect of nuclear casein and cyclin-dependent kinase substrate 1 (NUCKS1) gene on the proliferation of breast cancer cells and its association with different molecular subtypes of breast cancer. Methods TCGA and GTEx Databases were used to analyze the expression of NUCKS1 in tumor tissue of breast cancer patients and its association with prognosis. A total of 88 paraffin-embedded breast cancer specimens were collected from the patients who were diagnosed with breast cancer in Qingdao Central Hospital, University of Health and Rehabilitation Sciences, from January 2017 to December 2025, among whom there were 34 patients with triple-negative breast cancer (TNBC), 23 patients with HER2-overexpressing breast cancer, and 31 patients with luminal breast cancer. Paraffin-embedded tumor tissue specimens from all patients were used to construct tissue microarrays, and immunohistochemistry staining was used to measure the expression level of NUCKS1. The correlation between NUCKS1 expression and clinicopathological parameters was analyzed, as well as the difference in the expression of NUCKS1 across the different molecular subtypes of breast cancer. RT-qPCR and Western blot were used to mea-sure the mRNA and protein expression levels of NUCKS1 in five human breast cancer cell lines, i.e., MDA-MB-453, MDA-MB-231, MDA-MB-468, MCF-7, and T47D. MCF-7 cells were divided into groups A-D, while MDA-MB-231 cells were divided into groups a-d. The cells in groups A/a and B/b were transfected with empty pcDNA3.1 plasmid and NUCKS1-overexpressing pcDNA3.1 plasmid, respectively, and those in groups C/c and D/d were transfected with negative control lentivirus and CRISPR/Cas9-mediated NUCKS1-knockout lentivirus. CCK-8 assay was used to measure cell viability, and colony formation assay was used to evaluate the proliferative capacity of cells in each group. Results The bioinformatics analysis based on the TCGA and GTEx Databases showed that NUCKS1 was highly expressed in breast cancer tissue and was correlated with molecular subtypes, clinical stage, the expression of Ki-67, and the prognosis of patients (P<0.05). Immunohistochemistry staining showed that the expression level of NUCKS1 was significantly correlated with the percentage of Ki-67-positive cells (P<0.05). The patients with high NUCKS1 expression accounted for the highest proportion among the patients diagnosed with luminal breast cancer (χ²=6.37,P<0.05). RT-qPCR and Western blot showed that compared with the other breast cancer cell lines, MCF-7 cells showed the highest mRNA expression level of NUCKS1 (F=16.30,P<0.05), and MCF-7 and MDA-MB-231 cells showed the highest protein expression level of NUCKS1 (F=14.58,P<0.05), while there was no significant difference in the protein expression level of NUCKS1 between MCF-7 cells and MDA-MB-231 cells (P>0.05). CCK-8 assay showed that there was a significant difference in optical density between groups A and B and between groups a and b since 72 hours (P<0.05), and there was a significant difference in optical density between groups C and D and between groups c and d since 48 hours (P<0.05). Colony formation assay showed that group B had a significantly higher number of colonies than group A, and group C had a significantly higher number of colonies than group D (t=4.63,4.82,P<0.05); group b had a significantly higher number of colonies than group a, and group c had a significantly hig-her number of colonies than group d (t=3.23,6.62,P<0.05). Conclusion The expression level of NUCKS1 in luminal breast cancer is significantly higher than that in TNBC and HER2-overexpressing breast cancer. Overexpression of NUCKS1 can promote the proliferation of breast cancer cells, whereas NUCKS1 knockout significantly inhibits the proliferation of breast cancer cells.

Objective To investigate the effect of dihydroartemisinin on the adhesion, migration, invasion, and epithe-lial-mesenchymal transition (EMT) of human hepatocellular carcinoma HCCLM3 cells and to elucidate its mechanism. Methods Human hepatocellular carcinoma HCCLM3 cells were divided into groups A-D: group A (control group, group B (12.5 μmol/L dihydroartemisinin), group C (25.0 μmol/L dihydroartemisinin), group D (50.0 μmol/L dihydroartemisinin, group E [25.0 μmol/L dihydroartemisinin+50.0 μmol/L AG490, a Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway inhibitor], and group F (25.0 μmol/L dihydroartemisinin+0.5 μmol/L colivelin, a JAK2/STAT3 pathway activator). Cell viability, adhesion, migration, invasion, and the levels of related proteins were analyzed using CCK-8 assay, cell adhesion assay, Transwell assay, and Western blotting. Results Significant differences were observed among groups A-F in cell viability, adhesion, migration, invasion, and the expression levels of proteins related to the JAK2/STAT3 pathway and EMT (F=22.119-783.032,P<0.05). Groups B, C, and D showed reduced cell viability, adhesion, migration, invasion, and expression of N-cadhe-rin, vimentin, fibronectin and p-JAK2/JAK2, p-STAT3/STAT3 ratio compared with group A (t_LSD=2.451-43.655,P<0.05). In contrast, the expression level of E-cadherin in groups B, C, and D was higher than that in group A (t_LSD=30.671-85.442,P<0.05). Compared with group C, the changes in these indicators were more pronounced in group E (t_LSD=10.199-37.947,P<0.05), whereas group F showed the opposite trend (t_LSD=5.882-61.518,P<0.05). Conclusion Dihydroartemisinin significantly inhibits the adhesion, migration, invasion, and EMT of HCCLM3 cells, likely through suppression of the signal transduction of the JAK2/STAT3 pathway.

Objective To investigate the effect of scaffold protein DLGAP1 expression on the malignant phenotype of breast cancer cells and the underlying mechanism. Methods MDA-MB-231-mCherry (hereinafter referred to as 231) cells, a human triple-negative breast cancer cell line, were divided into the experimental group (siDLGAP1 group) and the negative control group (NC group). The 231 cells in the siDLGAP1 group were transfected with specific small interfering RNA targeting the DLGAP1 gene, while the 231 cells in the NC group were transfected with negative control small interfering RNA. The relative protein expression levels of DLGAP1, NR2B, and phosphorylated NR2B in the two groups of cells were detected by Western blotting assay. The proliferation, migration, and invasion abilities of the two groups of cells were determined by CCK-8 assay, wound healing assay, and Transwell assay, respectively. Results Western blotting assay showed that the relative expression levels of DLGAP1 and phosphorylated NR2B in the siDLGAP1 group were significantly lower than those in the NC group (t=4.70,17.46,P<0.01). The CCK-8, wound healing, and Transwell assays demonstrated that the proliferation, migration, and invasion abilities of cells in the siDLGAP1 group were significantly lower than those in the NC group (t=2.47-13.24,P<0.05). Conclusion Downregulation of scaffold protein DLGAP1 expression in breast cancer 231 cells can inhibit the malignant phenotype of these cells. The mec-hanism may be related to the regulation of the phosphorylation level of NR2B, suggesting that DLGAP1 may serve as a potential molecular target for breast cancer therapy.

Objective To construct a predictive model for the prognosis of patients with hepatocellular carcinoma (HCC) based on immunogenic cell death (ICD)-related genes, and to assess its clinical translational value. Methods A total of 365 HCC patients were selected from the Liver Hepatocellular Carcinoma (LIHC) cohort in The Cancer Genome Atlas database and were randomly divided into a training set and an internal validation set at a ratio of 7∶3, and a total of 108 HCC patients from the GSE76427 dataset in the Gene Expression Omnibus (GEO) database were selected as the external validation set. The bulk RNA-sequencing data of tumor tissue and the corresponding adjacent normal tissue were collected from the above patients, as well as related clinical data including sex, age, tumor grade, and overall survival. Single-cell RNA-sequencing data of 4 HCC patients were obtained from the GSE162616 dataset of the GEO database. Dimensionality reduction and clustering were performed for single cells from the GSE162616 dataset, and ICD activity scores were calcula-ted for each type of cells. Cells were divided into high ICD activity group and low ICD activity group according to ICD activity score, and differentially expressed genes (DEGs) between the two groups were identified. Weighted gene co-expression network analysis (WGCNA) was performed for the RNA-sequencing data of 365 HCC patients in the LIHC cohort to identify the gene set with the strongest association with ICD, and this gene set was intersected with the DEGs between HCC tumor tissue and the corresponding adjacent normal tissue of the patients in the LIHC cohort, finally obtaining the HCC-specific ICD-related genes (ICDRGs). The univariate Cox regression analysis was used to identify prognosis-related ICDRGs among the above genes; 10 machine learning algorithms were integrated to select the optimal genes for constructing an ICD-related HCC prognostic risk score model; the Spearman correlation analysis was used to investigate the correlation between the expression levels of these optimal genes and immune cell infiltration in HCC tissue. The model was used to calculate the prognostic risk score for each patient in the training set, the internal validation set, and the external validation set, and each set of patients were divided into high- and low-risk groups based on the median score. The Kaplan-Meier (K-M) survival analysis, the receiver operating characteristic (ROC) curve analysis, and the Cox regression analysis were used to evaluate the performance of the model in predicting prognosis. The univariate and multivariate Cox regression analyses were used to identify the influencing factors for the prognosis of HCC patients, which were integrated with the risk score to establish a nomogram model, and calibration curve and decision curve analysis (DCA) were used to assess the clinical translational value of the nomogram model. Results Five cell types were clustered in the GSE162616 dataset, among which immature B cells exhibited the highest ICD activity. A total of 920 ICD-related genes were identified at the single-cell level from the high and low ICD activity groups, and 82 HCC-specific ICDRGs were finally obtained through WGCNA. The univariate Cox regression analysis identified 26 prognosis-related ICDRGs from the above 82 genes, and after screening by machine learning algorithms, an ICD-related HCC prognostic risk score model was established based on 5 core genes, namely PIP4K2A, TAGLN2, RAC1, SLC16A3, and FYN. The Spearman correlation analysis showed that the expression levels of these 5 genes were significantly correlated with immune cell infiltration (P<0.001). The K-M survival analysis and the ROC curve analysis showed that this model had a good performance in predicting the prognosis of HCC patients, and the univariate and multivariate Cox regression analyses showed that the risk score of this model could be used as an independent predictive factor for the prognosis of HCC patients (HR=1.991,95%CI=1.368-2.898,P<0.001). The calibration curve showed that the nomogram model had a high accuracy in predicting the 1-, 3-, and 5-year survival rates of HCC patients, and DCA suggested that the nomogram model had the highest clinical benefit rate in terms of its performance in predicting 5-year survival rate. Conclusion The predictive model for the prognosis of HCC patients constructed in this study based on 5 HCC prognosis-related ICDRGs shows a good performance, and the nomogram model derived from the combination of this model and clinical indices also has a high clinical application value.

Objective To investigate the efficacy and safety of adalimumab in the treatment of children with different subtypes of non-systemic juvenile idiopathic arthritis (JIA). Methods A retrospective analysis was performed for the clinical data of 82 children with non-systemic JIA who were diagnosed and treated in The Affiliated Hospital of Qingdao University from July 2019 to June 2024. Based on the classification criteria established by the International League of Associations for Rheumatology, the children were divided into enthesitis-related arthritis (ERA) group with 45 children, oligoarticular JIA (oJIA) group with 18 children, rheumatoid factor-negative polyarticular JIA [RF(-)pJIA] group with 10 children, and rheumatoid factor-positive po-lyarticular JIA [RF(+)pJIA] group with 9 children. All children received adalimumab once every two weeks. The four groups children were compared in terms of peripheral blood C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), the number of active joints, and Juvenile Arthritis Disease Activity Score 27 (JADAS27) before treatment and at weeks 4, 12, and 24 of treatment. Results There was a significant difference in age of onset between the four groups (F=14.97,P<0.05), while there was no significant difference in sex distribution between the four groups (P>0.05). There was also a significant difference in affected joint sites between the four groups (χ²=9.96-16.80,P<0.05). Before treatment, and at different time points cluring treatment, the number of active joints were all statistically significant between the four groups (χ²=65.32-212.90,P<0.05); over the time of treatment, there was a tendency of reduction in the number of active joints in all four groups (χ²=76.40-241.50,P<0.05). There were no significant differences in peripheral blood CRP, ESR, and JADAS27 between the four groups before treatment (P>0.05), while there were significant differences in these indicators at different time points of treatment within each group, and all four groups had significant changes in these indicators after treatment (F=7.47-100.86,P<0.05). No adverse reactions such as local injection reactions, severe infections, liver/kidney dysfunction, and gastrointestinal discomfort were observed in any of the four groups during follow-up. Conclusion Adalimumab has a significant efficacy in the treatment of children with different subtypes of non-systemic JIA, although there is a difference in the number of active joints between the children with different subtypes at each time point of treatment. Adalimumab has a favorable short-term safety profile.

Objective To investigate the value of ARID1A mutation in tumor tissue in predicting immunotherapy response in gastric adenocarcinoma, and to establish a clinical predictive model. Methods A total of 777 patients with gastric adenocarcinoma from the cBioPortal for Cancer Genomics Database were selected as the exploratory cohort; 36 patients with gastric adenocarcinoma who received postoperative treatment with pembrolizumab monotherapy or pembrolizumab combined with chemotherapy in The Affiliated Hospital of Qingdao University were enrolled as validation cohort 1, and 93 patients with gastric adenocarcinoma from the cBioPortal for Cancer Genomics Database who were treated with immune checkpoint inhibitors (ICIs) were enrolled as validation cohort 2. According to the status of ARID1A mutation, the patients in each of the three cohorts were divided into mutation group and non-mutation group. The Kaplan-Meier method was used to plot survival curves, and the log-rank test and the univariate Cox proportional-hazards regression analysis were used to investigate the association between ARID1A mutation and prognosis in each cohort. In the validation cohort 1, immunohistochemistry was used to calculate CD8⁺/CD4⁺ T cell ratio in tumor tissue; for the patients in the validation cohort 2, the gradient boosting machine (GBM) algorithm was used to establish a GBM model, and multivariate Cox proportional-hazards regression was used to establish a Cox model. Index of concordance (C-index) and the area under the ROC curve (AUC) were used to assess the predictive performance of the two models, the calibration curve was used to assess the calibration of the two models, and the decision curve analysis was used to assess their clinical applicability. Results In the exploratory cohort, compared with the non-mutation group, the mutation group had a significantly higher incidence rate of microsatellite instability-high tumor (χ²=51.264,P<0.05) and a significantly longer progression-free survival (PFS) (HR=0.586, 95% CI=0.393-0.874,P<0.05). In the validation cohort 1, compared with the non-mutation group, the mutation group had a significantly longer PFS (HR=0.343,95%CI=0.156-0.751,P<0.05) and a significantly higher CD8⁺/CD4⁺ T cell ratio in tumor tissue (t=8.252,P<0.05). In the validation cohort 2, compared with the non-mutation group, the mutation group had a significantly higher tumor mutational burden (Z=3.113,P<0.05) and a significantly longer overall survival (HR=0.343,95%CI=0.156-0.751,P<0.05). Compared with the Cox model, the GBM model established based on the validation cohort 2 had better calibration and clinical applicability. Conclusion ARID1A mutation in tumor tissue has a favorable response to immunotherapy in patients with gastric adenocarcinoma, and the GBM model established based on the factors including ARID1A mutation has a good performance in predicting the prognosis of patients with gastric adenocarcinoma.

Objective To assess right ventricular systolic function in patients with complete left bundle branch block (CLBBB) and different levels of left ventricular ejection fraction (LVEF) using three-dimensional speckle-tracking imaging (3D-STI). Methods A total of 52 patients who were diagnosed with CLBBB in The Affiliated Hospital of Qingdao University from June 2024 to May 2025 were enrolled, and according to LVEF, they were divided into preserved LVEF group with 18 patients, mildly reduced LVEF group with 20 patients, and reduced LVEF group with 14 patients. A total of 35 healthy subjects during the same period of time were enrolled as control group. Two-dimensional echocardiography was used to measure the peak systolic velo-city of the tricuspid annulus (S'), tricuspid annular plane systolic excursion (TAPSE), fractional area change (FAC) of the right ventricle, right ventricular index of myocardial performance (RIMP), and 3D-STI was used to measure right ventricular free wall longitudinal strain (RVFWLS), right ventricular septal longitudinal strain (RVSLS), right ventricular ejection fraction (RVEF), right ventricular stroke volume (RVSV), right ventricular end-diastolic volume (RVEDV), right ventricular end-systolic volume (RVESV), right ventricular end-diastolic volume index (RVEDVi), right ventricular end-systolic volume index (RVESVi), and the right ventricular-pulmonary arterial coupling parameter RVFWLS/pulmonary artery systolic pressure (PASP) ratio. Two-dimensional echocardiography and 3D-STI parameters were compared between groups; the Spearman correlation analysis was used to investigate the correlation between right ventricular 3D-STI parameters and RVEF; the receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC) were used to assess the efficacy of 3D-STI parameters in the diagnosis of reduced right ventricular function in CLBBB patients. Results Two-dimensional echocardiography showed that there were significant diffe-rences in S', TAPSE, FAC, and RIMP between the four groups (F=4.607-68.009,P<0.01). Comparison of 3D-STI parameters showed that there were significant differences in RVESV, RVESVi, RVEF, RVSLS, RVFWLS, and RVFWLS/PASP ratio across the four groups (H=18.449-66.740,P<0.001); the three CLBBB groups had a significantly lower RVSLS than the control group (P<0.05); the reduced LVEF group had significantly lower RVEF RVFWLS, and RVFWLS/PASP ratio than the other three groups, a significantly lower RVSLS than the preserved LVEF group, and significantly higher RVESV and RVESVi than the control group and the preserved LVEF group (P<0.05); compared with the control group, the mildly reduced LVEF group had a significantly lower RVEF and a significantly higher RVFWLS/PASP ratio (P<0.05). The Spearman correlation analysis showed that the 3D-STI parameters RVSLS, RVFWLS, and RVFWLS/PASP ratio were significantly negatively correlated with RVEF (r=-0.698--0.626,P<0.001). The ROC curve analysis showed that 3D-STI parameters had relatively high efficacy in asses-sing right ventricular systolic dysfunction in patients with CLBBB, with an AUC of 0.915. Conclusion This study shows that 3D-STI parameters can effectively assess the change in right ventricular systolic function in patients with CLBBB and thus provide reliable imaging data for the early identification of right ventricular systolic dysfunction in clinical practice.

Objective To analyze the clinical characteristics of external auditory canal and temporal bone hemangiomas, and to improve the diagnosis and treatment of these conditions. Methods A retrospective analysis was conducted on the clinical data of six patients with external auditory canal hemangiomas and one patient with temporal bone hemangioma. Their clinical features and diagnostic and treatment processes were summarized. Results External auditory canal and temporal bone hemangiomas are relatively rare, typically presenting with symptoms such as otorrhea, hearing loss, tinnitus, and aural fullness. Magnetic resonance imaging showed isointensity on T₁-weighted imaging and hyperintensity on T₂-weighted imaging in the lesions. All seven patients underwent surgical treatment, and the diagnosis was confirmed by postoperative pathological examination in all cases. At 1-3 months postoperatively, follow-up examination revealed satisfactory wound healing and no recurrence. At the 6-month follow-up, none of the seven patients reported any significant discomfort. Conclusion External auditory canal and temporal bone hemangiomas are clinically rare and have atypical clinical features. The characteristic magnetic resonance imaging findings (isointensity on T₁-weighted imaging and hyperintensity on T₂-weighted imaging) are of great value for differential diagnosis and preoperative assessment. Surgical approaches should be selected on an individualized basis according to the location and extent of the lesion, and standardized diagnosis and treatment can effectively improve patient outcomes.

Objective To investigate the efficacy and safety of tirofiban in the treatment of acute progressive cerebral infarction (APCI). Methods A total of 88 patients with APCI were enrolled as subjects, and they were divided into treatment group and control group using a random number table, with 44 patients in each group. The patients in the control group received dual antiplatelet therapy (aspirin and clopidogrel) combined with conventional symptomatic treatment, and those in the treatment group received continuous intravenous infusion of tirofiban for 48 h, followed by dual antiplatelet therapy and conventional symptomatic treatment. The two groups were assessed in terms of National Institutes of Health Stroke Scale (NIHSS) score and Barthel Index (BI) before treatment and on day 14 of treatment; the indicators of coagulation and platelets were measured and compared between the two groups, including platelet count (PLT), prothrombin time (PT), and platelet aggregation rate (PAgT) before treatment, at 48 hours of treatment, and on day 14 of treatment. Results Before treatment, there were no significant differences in NIHSS score and BI between the two groups (P>0.05), and on day 14 of treatment, both groups had a significant reduction in NIHSS score and a significant increase in BI (t=15.03-14.95,P<0.05); there were significant differences between the two groups in the changes in NIHSS score and BI after treatment (t=2.45, 2.68,P<0.05). Group, time, and group-time interaction had a significant impact on PAgT (F_time=132.45,F_group=42.18,F_interaction=95.67,P<0.05), while they had no significant impact on PT and PLT (P>0.05). For the treatment group, PAgT level at 48 hours of treatment was significantly lower than that before treatment and on day 14 of treatment (P<0.05), and there was no significant difference between PAgT level on day 14 of treatment and that before treatment (P>0.05). Conclusion Tirofiban has good efficacy in the treatment of APCI and can effectively improve neurological deficits and enhance the activities of daily living, with a manageable safety profile.

Objective To investigate the risk factors for gastrointestinal bleeding (GIB) in patients with chronic kidney disease (CKD). Methods A total of 335 CKD patients who were admitted to The Affiliated Hospital of Qingdao University from April 2023 to October 2025 were enrolled, and according to the presence or absence of GIB, they were divided into GIB group with 188 patients and non-GIB group with 147 patients. The two groups were compared in terms of age, sex, chronic diseases (hypertension, liver cirrhosis, diabetes, and cardiovascular and cerebrovascular diseases), medication (antiplatelet agents, anticoagulants, and hormones), history of dialysis treatment, estimated glomerular filtration rate (eGFR), serum creatinine, blood urea nitrogen, and serum albumin. Univariate and multivariate logistic regression analyses were used to identify the risk factors for GIB in CKD patients. The receiver operating characteristic (ROC) curve was used to evaluate the efficacy of statistically significant variables in the diagnosis of GIB. Results There were significant differences between the GIB group and the non-GIB group in age, sex, eGFR, history of cardiovascular and cerebrovascular diseases, history of diabetes serum, albumin level, and history of antiplatelet drug use (χ²=4.458-40.964,Z=-4.443,t=-5.499,P<0.05). The multivariate logistic regression analysis showed that old age, male sex, a reduction in eGFR, a reduction in serum albumin level, and a history of antiplatelet drug use were independent risk factors for GIB in CKD patients (P<0.05). The ROC curve analysis showed that the combination of age, sex, eGFR, serum albumin, and history of antiplatelet drug use yielded an area under the ROC curve (AUC) of 0.800 for predicting GIB. Conclusion Old age, male sex, a reduction in eGFR, hypoalbuminemia, and history of antiplatelet drug use are independent risk factors for GIB in CKD patients. The predictive model combining multiple indicators has good predictive performance, and multidimensional comprehensive assessments and individualized interventions should be performed in clinical practice to reduce the risk of GIB and improve the prognosis of patients.

Objective To investigate the clinical phenotype and genetic characteristics of a child with cardio-facio-cuta-neous syndrome (CFCS), and to evaluate the efficacy and safety of growth hormone therapy through a literature review. Methods Clinical data were collected from a child with CFCS and poor postnatal weight gain, and after confirmed diagnosis by whole-exome sequencing (WES), the patient was treated with long-acting recombinant human growth hormone (LA-rhGH) for one year. Growth parameters and safety indicators were closely monitored. A systematic literature review was performed for the reports of CFCS. Results A girl, aged 1 year and 4 months, presented with feeding difficulties and growth retardation after birth, with unsatisfactory improvement in growth after nutrition guidance and nutrient supplementation. The girl also had unusual facies, skin and hair abnormalities, and atrial septal defect. Genetic testing identified a de novo heterozygous missense mutation, c.1785T>G (p.F595L), in the BRAF gene, and thus the girl was diagnosed with CFCS. The girl received regular treatment with LA-rhGH at a dose of 0.2 mg/(kg·week) for one year. The annual height growth rate reached 12.5 cm/year; height Z-score was improved from -3.2 to -2.1, weight Z-score was improved from -2.8 to -1.9, and the serum level of insulin-like growth factor-1 (IGF-1) was increased from <15 μg/L before treatment to 96.8 μg/L. Follow-up during and after treatment showed that liver and kidney function parameters, blood glucose, thyroid function parameters, and tumor markers all remained within the normal range, and there were no other adverse effects. Conclusion For children with CFCS caused by a mutation in the BRAF gene, rhGH therapy can effectively promote the linear growth of body height and increase the serum level of IGF-1, with a favorable safety profile for short-term application. But long-term follow-up is needed to clarify its risk benefitratio.

Objective To construct an interpretable machine learning model based on Delta radiomic features of intratumoral and peritumoral regions on multi-parametric MRI, and to evaluate its performance in predicting pathological complete response (pCR) after neoadjuvant therapy (NAT) in patients with breast cancer. Methods A total of 183 patients from The Affi-liated Hospital of Qingdao University were enrolled and randomly divided into a training set with 128 patients and an internal validation set with 55 patients, and 71 patients from Yantai Yuhuangding Hospital were enrolled as the external validation set. Related clinicopathological data were collected from all patients, including age, menopausal status, axillary lymph node metastasis, and the expression of estrogen receptor (ER), progesterone receptor (PR), and Ki-67. Multi-parametric MRI data were collected from all patients before NAT and after two NAT cycles; intratumoral regions of interest (ROIs) were delineated, and different peritumoral ROIs were obtained by expanding the intratumoral boundaries by 3 mm, 5 mm, and 7 mm, respectively. The Pyradiomics package was used to extract the radiomic features of the above ROIs, and the values of delta radiomic features were calculated. The Extra Trees algorithm was used to construct four delta radiomics models (Delta_intratumoral, Delta_{intra+3 mm peritumoral}, Delta_{intra+5 mm peritumoral}, and Delta_{intra+7 mm peritumoral}). The receiver operating characteristic (ROC) curve was used to assess the predictive performance of each model; the decision curve analysis (DCA) was used to evaluate the clinical applicability of each model; the calibration curve and the Hosmer-Lemeshow goodness-of-fit test were used to assess the calibration degree of each model. Finally, the SHapley Additive exPlanations (SHAP) algorithm was used to interpret the optimal predictive model. Results There were no significant differences in age, menopausal status, axillary lymph node metastasis, and the expression of ER, PR, and Ki-67 between the training set, the internal validation set, and the external validation set (P>0.05). The ROC curve analysis and the DCA evaluation showed that the Delta_{intra+7 mm peritumoral} model yielded the highest AUC and net clinical benefit compared with the other three models in the external validation set. The calibration curve analysis showed that the Delta_{intra+7 mm peritumoral} model had the lowest Brier score compared with the other three models in the external validation set. The Hosmer-Lemeshow goodness-of-fit test showed good calibration for the Delta_{intra+7 mm peritumoral} model in the external validation set (P=0.074). The SHAP-based interpretability analysis of the Delta_{intra+7 mm peritumoral} model showed that among the morphological features extracted from ADC and contrast-enhanced MRI images, flatness and sphericity had a marked influence on the output of the Delta_{intra+7 mm peritumoral} model. Conclusion The interpretable machine learning models based on Delta radiomic features of intratumoral and peritumoral regions have a better predictive performance than the model based on intratumoral features alone in predicting pCR after NAT in breast cancer patients, and the Delta model based on intratumoral and 7 mm peritumoral regions has the best predictive performance and clinical applicability.

Sleep disorders may induce or exacerbate epilepsy, and seizures may interfere with the status of sleep. There are complex mechanisms for the comorbidity between sleep disorders (such as insomnia, obstructive sleep apnea, narcolepsy, and restless legs syndrome) and epilepsy; narcolepsy and epilepsy may share the same pathogenesis, and non-rapid eye movement sleep-related parasomnias and sleep-related hypermotor epilepsy need to be differentiated from each other. This article describes the mechanism of the relationship between epilepsy and sleep disorders and elaborates on the influence of sleep architecture on epilepsy, in order to provide a reference for effective seizure control and the treatment of sleep disorders in such patients.