Objective: To evaluate the predictive value of brain structural network characteristics for the risk of depression in patients with sleep disorders. Methods: A two-year prospective follow-up was performed in 129 patients with sleep disorders enrolled from Outpatient Departments of Neurology and Otolaryngology, Affiliated Yixing Hospital of Jiangsu University from January 2020 to December 2021. A total of 50 age- and gender-matched healthy controls were recruited by local community advertisements at the same time period. All subjects underwent 3.0 T MRI and brain structural networks were constructed. They were then followed up for 2 years. During the period, all patients were assessed for depression occurrence by Hamilton Depression Scale (HAMD) and divided into group of sleep disorders with depression and group of sleep disorders without depression accordingly. Differences in baseline information and brain structural networks were compared among the 3 groups, and independent influencing factors for depression in patients with sleep disorders were analyzed by multivariate Logistic regression. The predictive value of brain structural network indexes in depression in patients with sleep disorders was analyzed by receiver operator characteristic (ROC) curve. Partial correlation analysis was conducted to explore the correlation between brain structural network indexes and HAMD scores in patients with sleep disorders. Results: Compared with the healthy control group, the group of sleep disorders without depression and group of sleep disorders with depression had significantly lower global efficiency, and node efficiency of the left amygdala, right fusiform gyrus, left superior frontal gyrus and left hippocampus (all P<0.05). Compared with the group of sleep disorders without depression, the group of sleep disorders with depression had significantly lower global efficiency, statistically higher clustering coefficient, and significantly lower node efficiency of the left hippocampus, left amygdala and right superior occipital gyrus (all P<0.05). Logistic regression analysis revealed that global efficiency, nodal efficiency of the left hippocampus, and nodal efficiency of the left amygdala were independent influencing factors for depression in patients with sleep disorders(P<0.05). The ROC curve analysis results indicated that the area under the curve for predicting depression in patients with sleep disorders by the regression model constructed with the above three factors was 0.882 (95%CI: 0.815-0.953, P<0.01). Partial correlation analysis indicated that global efficiency, nodal efficiency of the left hippocampus, and nodal efficiency of the left amygdala in the group of sleep disorders with depression were negatively correlated with HAMD scores at the end of follow-up (r=-0-.672, -0.618, -0.649, all P<0.01). Conclusion: Baseline global efficiency, nodal efficiency of the left hippocampus, and nodal efficiency of the left amygdala could be used to effectively predict the risk of depression in patients with sleep disorders and are closely associated with depression severity.

Objective: To establish a optogenetic model of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) activation in the hippocampal neurons of Alzheimer′s disease (AD) mice. Methods: Twentyfour APP/PS1 double transgenic (AD model) mice were selected and randomly divided into 4 groups based on the type of viral intervention (AAV-Ppargc1α-hChR2/AAV-Control-hChR2) and light treatment (light stimulation/no light stimulation): AD+AAV-Control-hChR2+no light stimulation group, AD+AAV-Control-hChR2+light stimulation group, AD+AAV-Ppargc1α-hChR2+no light stimulation group, and AD+AAV-Ppargc1α-hChR2+light stimulation group, with 6 in each group. Genotypes were confirmed by PCR. Adeno-associated virus carrying the lightsensitive protein hChR2 was microinjected into the dentate gyrus of the hippocampus of AD mice, and optical fibers were implanted at the injection site. Two weeks after viral expression, blue light stimulation was applied at 5-minute intervals lasting for 30 seconds each, for 2 hours a day, for 5 days. Immunofluorescence was used to detect the number of Cfos+HA+ cells in the hippocampus of the four groups of mice to verify the effectiveness of the lightsensitive protein; immunofluorescence was also used to detect the number of PGC-1α positive neurons in the hippocampus of AD mice under the same light stimulation conditions. Results: PCR results showed that mice with both APP and PS1 bands were AD double transgenic mice. Compared with the no light stimulation groups (AD+AAV-Control-hChR2+no light stimulation and AD+AAV-Pgc-1α-hChR2+no light stimulation), the number of Cfos+HA+ neurons in the hippocampus of the light stimulation groups (AD+AAV-Control-hChR2+light stimulation and AD+AAV-Pgc-1α-hChR2+light stimulation) increased significantly (P<0.01). Compared with AAV-Control-hChR2+light stimulation group, the expression of PGC-1α positive neurons in the hippocampus of AD mice in AAV-Pgc-1α-hChR2+light stimulation group was significantly increased (t=9.613, P<0.01). Conclusion: A hippocampus-specific optogenetic model for PGC-1α activation in AD mice was successfully established.

Objective: To investigate the effect of recombinant avirulent Newcastle disease virus LaSota strain expressing the rabies virus glycoprotein (rL-RVG) on ferritinophagy in gastric cancer cells and its potential mechanism. Methods: The CCK-8 assay was used to evaluate the effect of different concentrations of rL-RVG on the proliferation ablilty of gastric cancer HGC-27 and SGC-7901 cells, and the optimal concentration was screened out. Protein expression levels of adenosine monophosphate activated protein kinase (AMPK) , phosphorylated AMPK (p-AMPK), and Beclin-1, as well as nuclear receptor coactivator 4 protein(NCOA4), ferritin heavy chain (FTH1), and microtubuleassociated protein 1A/1B-light chain 3 (LC-3) in gastric cancer HCC-27 and SGC-7901 cells were detected by Western blotting. Meanwhile, the content of cellular Fe²⁺ was detected by cellular Fe²⁺ fluorescence assay kit. Gastric cancer HGC-27 and SGC-7901 cells were treated with siRNA-beclin-1 and the AMPK inhibitor Compound C. The effects of rL-RVG on ferritinophagy in gastric cancer cells were assessed by measuring the protein expression levels of the AMPK/Beclin-1 axis through Western blotting and the fluorescence intensity of Fe²⁺ using a cellular Fe²⁺ fluorescence assay kit. Gastric cancer HGC-27 and SGC-7901 cells were treated with rL-RVG diluted 10³ times, and the cell proliferation ability was determined by the CCK-8 method; Transwell invasion assay was used to detect the invasion ability of cells; scratch test was used to detect the migration ability of cells. Results: rL-RVG had a significant inhibitory effect on gastric cancer HGC-27 and SGC-7901 cells, and the inhibitory ability gradually increased with the decrease of virus dilution factor (P<0.05), when the virus dilution factor was 103 times, it was closest to the half inhibitory concentration (IC₅₀), therefore, rL-RVG diluted 10³ times was used for the subsequent experiments. rL-RVG significantly increased AMPK, p-AMPK, Beclin-1, LC-3 and NCOA4 protein expression, fluorescence intensity of Fe²⁺, and decreased FTH1 expression in gastric cancer HGC-27 and SGC-7901 cells (all P<0.05). Knockdown of Beclin-1 and inhibition of AMPK significantly decreased AMPK, p-AMPK, Beclin-1, LC-3 and NCOA4 protein expression, fluorescence intensity of Fe²⁺, and increased FTH1 expression in gastric cancer HGC-27 and SGC-7901 cells (all P<0.05). The addition of rL-RVG with a dilution of 10³ significantly inhibited the proliferation, invasion and migration of gastric cancer HGC-27 and SGC-7901 cells, while Compound C partially reversed the inhibitory effect of rL-RVG on gastric cancer cells (all P<0.05). Conclusion: rL-RVG could promote ferritinophagy in gastric cancer HGC-27 and SGC-7901 cells by affecting the expression of related proteins through AMPK/Beclin-1 axis.

Objective: To compare the effects of stem cell lysate prepared by repeated freeze-thaw method and ultrasonic disruption method on skin injury repair, and screen the optimal preparation method of stem cell lysate. Methods: Human skin fibroblasts were resuspended in normal saline at four densities (1×10⁵/mL, 1×10⁶/mL, 5×10⁶/mL, 1×10⁷/mL), and treated by repeated freezethaw and ultrasonic disruption, respectively. The cell disruption rate and protein content were detected by BCA assay to screen the optimal preparation conditions of the two methods. The scratch healing ability, viability and migration number of human skin fibroblasts were detected by cell scratch healing assay, CCK-8 method and Transwell assay, respectively. The protein concentration of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF) and stem cell factor (SCF) in the lysate were detected by ELISA. Results: The optimal condition of ultrasonic disruption method was cell concentration of 5×106/mL with ultrasonic treatment for 5 min; the optimal condition of repeated freezethaw method was cell concentration of 5×106/mL with three freeze-thaw cycles (45 min). There was no significant difference in cell disruption rate between the two methods (P>0.05), and the total protein content of lysate prepared by ultrasonic disruption method was higher. There was no significant difference in the effect of lysates prepared by the two methods on the viability of human skin fibroblasts (P>0.05). Compared with ultrasonic disruption method, the lysate prepared by repeated freezethaw method could significantly promote the migration of human skin fibroblasts and improve the efficiency of cell scratch repair (P<0.05). ELISA results showed that the concentrations of VEGF and SCF proteins in the lysate prepared by repeated freeze-thaw method were significantly higher than those by ultrasonic disruption method (P<0.05 or P<0.01), and there was no significant difference in EGF content (P>0.05). Conclusion: The stem cell lysate prepared by repeated freeze-thaw method has a better effect on skin injury repair than that by ultrasonic disruption method.

Objective: To explore the effect and mechanism of circGFRA1 on the proliferation, migration and invasion of colorectal cancer (CRC) SW620 cells. Methods: qRT-PCR was used to detect the expression levels of circGFRA1 and miR-3194-3p in CRC tissues, adjacent normal tissues, SW620 cells and normal human colonic epithelial NCM460 cells. SW620 cells were cultured in vitro and divided into 6 groups: si-NC group, si-circGFRA1 group, miR-NC group, miR-3194-3p group, si-circGFRA1+anti-miR-NC group, and sicircGFRA1+anti-miR-3194-3p (si-circGFRA1+anti-miR) group. CCK-8 assay and colony formation assay were used to detect cell proliferation ability; scratch assay and Transwell assay were used to detect cell migration and invasion abilities; dual luciferase reporter assay was used to verify the targeting relationship between circGFRA1 and miR31943p; Western blotting was used to detect the protein expression levels of MMP-2 and MMP-9. Results: The expression level of circGFRA1 in CRC tissues was significantly higher than that in adjacent normal tissues (P＜0.001), and the expression level of miR-3194-3p was significantly lower than that in adjacent normal tissues (P＜0.001); the expression level of circGFRA1 in SW620 cells was significantly higher than that in NCM460 cells (P＜0.001), and the expression level of miR-3194-3p was significantly lower than that in NCM460 cells (P＜0.001). Compared with the siNC group, the cell viability, number of colony formation, migration and invasion cells in the si-circGFRA1 group were significantly reduced (P＜0.05), and the protein expression levels of MMP-2 and MMP9 were significantly downregulated (P＜0.05); compared with the miR-NC group, the change trends of the above indicators in the miR-3194-3p group were consistent with those in the si-circGFRA1 group (P＜0.05). Dual luciferase reporter assay confirmed that circGFRA1 could specifically bind to miR-3194-3p. Compared with the si-circGFRA1+anti-miR-NC group, the cell viability, number of colony formation, migration and invasion cells in the si-circGFRA1+anti-miR group were significantly increased (P＜0.05), and the protein expression levels of MMP-2 and MMP-9 were significantly upregulated (P＜0.05). Conclusion: circGFRA1 is highly expressed in CRC, and promotes the proliferation, migration and invasion of CRC cells by targeting and inhibiting the expression of miR-3194-3p and upregulating the protein levels of MMP-2 and MMP-9.

Objective: To explore the risk factors affecting the prognosis of elderly patients with clear cell renal cell carcinoma (ccRCC) and construct a novel prognostic model. Methods: Within Surveillance, Epidemiology, and End Results （SEER） registry database, 10 236 elderly ccRCC patients were randomly assigned to the training groups and validation groups at a ratio of 7∶3. Multivariate Cox regression analysis was used to identify independent risk factors for the prognosis of elderly ccRCC patients. A prognostic model was constructed based on the regression coefficients of each factor in the Cox regression analysis, and a nomogram was established to predict the overall survival (OS), and cancer specific survival (CSS) at 1, 3, and 5 years in elderly ccRCC patients. The model were evaluated by receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis. The model was validated using data from the validation groups. Results: Multivariate Cox regression analysis showed that age, gender, race, marital status, tumor size, histological grade, T stage, N stage, M stage, surgical method, lymph node dissection, venous infiltration and metastatic site were independent risk factors for OS in elderly ccRCC patients (all P<0.05). In contrast, the independent risk factors for CSS included age, tumor size, histological grade, pathological stage, N stage, M stage, surgical method, venous infiltration and metastatic site (all P<0.05). The ROC curve of the OS model showed that the AUC values for 1, 3 and 5 year OS were 0.75, 0.74 and 0.72 in the training groups, and 0.74, 0.74 and 0.72 in the validation groups, respectively. For the CSS nomogram, the AUC values for 1, 3 and 5 year OS were 0.84, 0.84 and 0.82 in the training groups, and 0.83, 0.84 and 0.81 in the validation groups. The calibration curve showed that the nomogram predicted patient survival was highly consistent with actual survival. The clinical decision curve showed a higher net clinical benefit in the prognostic model than the TNM model. Conclusion: The nomogram model for predicting 1, 3 and 5 year OS and CSS in elderly ccRCC patients has good discriminative ability, model fit, and clinical net benefit.

Objective: To construct a quantitative index DL-ILD for interstitial lung disease (ILD) based on deep learning (DL), and to explore its clinical value in predicting the progression of ILD. Methods: A retrospective study was conducted on 112 patients with ILD. A three-dimensional U-Net (U-shaped convolutional neural network) model was constructed based on high resolution computerized tomography (HRCT) to automatically identify lesions and calculate DL-ILD. The evaluation criterion was whether ILD had progressed after a 24-month follow-up. The correlation between DL-ILD, visual scores, and lung function was assessed, and the predictive performance of the two methods for predicting progression and prognosis was compared. Logistic regression, receiver operating characteristic (ROC) curve analysis, Cox analysis, and fivefold crossvalidation were used to validate the model. Results: Among the 112 ILD patients, 42 experienced progression events. DLILD showed a negative correlation with forced vital capacity (FVC%) and diffusion capacity of the lung for carbon monoxide (DLCO%) (r=-0.762, r=-0.685, P＜0001), outperforming visual scores (Z=2.593, P=0.010; Z=2.598, P=0.009). Logistic regression analysis indicated that DL-ILD was an independent predictor of ILD progression (OR=1.242, P=0.001), with its predictive performance (AUC=0.864) superior to that of visual scores (AUC=0.710). The 1-year and 2-year progression-free survival rates of the high DL.ILD group were 82.1% and 53.7%, respectively, which were significantly lower than 98.4% and 84.6% of the low DL-ILD group (P<0.01), and the Cox model confirmed it as an independent risk factor for disease progression (HR=2.872, P=0.002). The AUC of the five-fold cross-validation was 0.833, indicating stable model performance. Conclusion: DL-ILD can accurately reflect the prediction of ILD disease progression, outperforming traditional visual scores, and has good reproducibility and clinical application potential.

Objective: To examine the impact of cognitive impairment on the reliability of visual field assessments in glaucoma patients, and to establish and validate a predictive model utilizing Mini-Cog Scores for early warning. Methods: A retrospective analysis was conducted on clinical data from 306 glaucoma patients treated at our institution between January 2022 and December 2024, forming the training cohort. Patients meeting reliability criteria (fixation loss ≤20%, false positive rate ≤15%, false negative rate ≤33%) were assigned to the study group (n=209), while those failing to meet these benchmarks constituted the control group (n=97). Inter-group comparisons of clinical characteristics were performed, with Logistic regression analysis employed to identify determinants of visual field reliability. Predictive capabilities of individual factors were evaluated through ROC curve analysis. An additional validation cohort comprised 131 glaucoma patients treated from January to June 2025, selected using the same criteria. A nomogram was developed via the Storm Statistics platform, with its predictive accuracy and clinical applicability assessed through calibration curves, ROC curve analysis, and decision curve evaluation. Results: The age and disease duration of the control group were significantly higher than those of the study group, while the best corrected visual acuity and Mini-Cog Score were significantly lower than those of the study group (all P<0.01). Multivariate Logistic regression identified age and disease duration as independent risk factors compromising visual field reliability, whereas best corrected visual acuity and Mini-Cog Scores emerged as independent protective factors (all P<0.05). Mini-Cog Score demonstrated superior predictive value relative to age, disease duration, and best corrected visual acuity (P<0.01). The constructed nomogram exhibited high predictive probability for visual field reliability, with AUC values of 0.89 (95%CI: 0.85-0.94) in the training set and 0.90 (95%CI: 0.82-0.98) in the validation set, indicating robust predictive performance. Calibration curves confirmed strong concordance between predicted probabilities and observed outcomes (P>0.05). Decision curve analysis revealed substantial clinical net benefit when threshold probabilities ranged from 30% to 90%. Conclusion: Cognitive impairment substantially compromises the reliability of visual field testing in glaucoma patients. The early warning model incorporating Mini-Cog Scores demonstrates favorable predictive capability and practical clinical value.

Objective: To evaluate the association between cardiac rehabilitation (CR) compliance and the improvement of left ventricular ejection fraction (LVEF) as well as cardiovascular readmission within 1 year in patients with multivessel coronary artery disease (CAD) after percutaneous coronary intervention (PCI), and to explore the effect heterogeneity in different subgroups. Methods: This was a singlecenter retrospective cohort study. A total of 200 patients with multivessel CAD who underwent PCI and had complete followup data in Huai′an Rehabilitation Hospital from December 2023 to December 2024 were enrolled. According to the completion rate of CR program, the patients were divided into the high compliance group (≥70%, n=70), moderate compliance group (40%-69%, n=60) and low compliance group (<40% or non-participation, n=70). The primary endpoints were LVEF improvement (ΔLVEF>0) during followup and cardiovascular readmission within 1 year. Multivariate Logistic regression analysis was used to identify independent associations, and subgroup and interaction analyses were performed by age, gender, diabetes status and revascularization status. Results: The baseline demographic and clinical characteristics were generally balanced among the three groups. The proportion of incomplete revascularization in the low compliance group was significantly higher than that in the high compliance group (70.0% vs. 50.0%, P=0.034), while the proportion of good medication compliance was significantly lower (45.7% vs. 90.0%, P<0.001). There were statistically significant differences in the incidence of LVEF improvement (70.0%, 50.0%, 30.0%) and 1-year cardiovascular readmission rate (14.3%, 2..0%, 35.7%) among the high, moderate and low compliance groups (both P<0.05). Multivariate analysis showed that compared with the high compliance group, the moderate compliance group (OR=0.52, 95%CI: 0.28-098, P=0041) and low compliance group (OR=0.30, 95%CI: 0.16-0.57, P<0.001) had a significantly lower probability of LVEF improvement, and the risk of readmission increased by 1.92 times (OR=1.92, 95%CI: 1.03-3.59, P=0.039) and 3.25 times (OR=3.25, 95%CI: 1.76-6.01, P<0.001), respectively. Incomplete revascularization (P=0.013), poor medication compliance (P=0.005) and baseline LVEF<50% (P=0.007) were independent predictors of adverse outcomes. Subgroup analysis indicated that the beneficial effect of CR compliance was more pronounced in patients aged ≥65 years and those with incomplete revascularization, and there was a significant interaction between revascularization status and CR compliance (P=0.040). Conclusion: In patients with multivessel CAD after PCI, CR compliance has an independent and dose-dependent association with LVEF improvement and reduced risk of cardiovascular readmission. A comprehensive intervention approach combining complete revascularization and good medication compliance can maximize the clinical benefits of CR. It is particularly recommended to strengthen the management of CR compliance in elderly patients and those with incomplete revascularization.

Objective: To construct a novel argininefunctionalized metal-polyphenol nanopolymer (Man-Fe@Arg) and investigate its killing effect and mechanism in tumor chemodynamic therapy (CDT). Methods: Aldehyde-modified mangiferin was prepared by oxidation method, and arginine was grafted via Schiff base reaction; Man-Fe@Arg was synthesized by metal-polyphenol coordination self-assembly method. The physicochemical properties were characterized by scanning electron microscopy, Malvern particle size analyzer, Zeta potential analyzer and Fourier transform infrared spectroscopy. The Fenton catalytic activity, cellular nitric oxide (NO) secretion level, cellular uptake capacity, biocompatibility and killing effect on mouse breast cancer 4T1 cells were evaluated by in vitro experiments. Results: The successfully prepared Man-Fe@Arg had an average particle size about 157.3 nm with good dispersibility and excellent biocompatibility. The introduction of arginine could significantly increase the NO release level in tumor cells, strengthen the Fenton reaction through the synergistic effect of NO and reactive oxygen species, and greatly improve the killing efficiency of CDT on 4T1 cells. Conclusion: Man-Fe@Arg could significantly improve the killing effect of CDT through arginine-enhanced Fenton reaction.