ObjectiveThis paper aims to conduct the feature analysis and correlation analysis on the ocular collateral features and differential metabolites in patients with chronic hepatitis B （CHB） complicated by glucose metabolism disorder （GMD），particularly those with the damp-heat syndrome type，by integrating shadowless scleral imaging and metabolomics technologies. MethodsA total of 313 patients were recruited from the Hepatology and Endocrinology Outpatient Departments of Public Health Clinical Center of Chengdu according to the inclusion/exclusion criteria，and they were divided into a CHB group and a CHB complicated by GMD groups （damp-heat syndrome group and non-damp-heat syndrome group）. All patients underwent high-definition ocular image acquisition and feature extraction using an intelligent analysis system for shadowless scleral imaging to analyze the differences in the counting of morphological feature scores of ocular collaterals among groups. By using a digital sampling method，24 patients from each group were randomly selected，along with 20 healthy volunteers，for untargeted metabolomic analysis of peripheral serum. Differential metabolites were identified，statistically analyzed，and subjected to potential biomarker analysis and pathway enrichment. Spearman method was performed to conduct the correlation analysis on the differential ocular collateral features and differential metabolites，followed by correlation network construction. ResultsCompared with those in the CHB group，patients with CHB complicated by GMD showed significant changes in ocular collateral feature scores such as "hillock"，"blood vessels"，and "pale dusky coloration" （P<0.05）. In comparison with those in the healthy group，metabolites including N-acetylglucosamine，acetylhomoserine，and myo-inositol （AUC>0.7） were identified as potential biomarkers for the disease. Compared with those in the CHB complicated by GMD group with non-damp-heat syndrome，patients with damp-heat syndrome exhibited significant changes in feature scores of "plaques"，"yellow coloration"，"spleen"，and "gallbladder" （P<0.05）. In comparison with those in the healthy group，metabolites such as O²′-4a-cyclic tetrahydrobiopterin，theobromine，xanthurenic acid，and L-glutamic acid 5-phosphate （AUC>0.7） were identified as potential biomarkers for the damp-heat syndrome type. The Spearman correlation analysis reveals weak to moderate linear correlations between the differential scleral collateral features and metabolites. By constructing a "disease-syndrome" network of ocular diagnosis and metabolites，"xanthurenic acid-gallbladder" and "theobromine-plaque/yellow coloration" were identified as specific molecular-phenotypic correlated biomarker clusters for CHB complicated by GMD with dampness-heat syndrome. ConclusionPatients with CHB complicated by GMD demonstrate differential ocular diagnostic features and serum metabolites corresponding to disease states and dampness-heat syndrome. These objective biomarkers can guide both clinical syndrome differentiation and medication. The macro-micro integration based on ocular feature clusters and potential metabolic biomarkers offers an innovative approach to a combined traditional Chinese and Western medicine diagnosis and treatment model for this disease.

AIM:To explore the central mechanisms by which metformin(Met)attenuates insulin resistance in high-fat diet(HF)-fed rats.METHODS:Forty healthy male Sprague-Dawley rats were randomly divided into 4 groups:normal chow(NC)group,HF group,HF+Met group,and HF+Met+SHU9119[melanocortin 4 receptor(MC4R)antagonist]group,with 10 rats per group.Treatments with HF and Met lasted for 12 weeks,while SHU9119 was injected for the last 10 d.Skeletal muscle AMP-activated protein kinase(AMPK)and silent information regulator 1(SIRT1)ex-pression and activity were measured,along with mitochondria oxidative stress markers,mitochondrial function and quanti-ty.Systemic and skeletal muscle insulin sensitivity were assessed using the average glucose infusion rate from 60 to 120 min(GIR60-120)and 2-deoxyglucose uptake(DGU)during hyperinsulinemic-euglycemic clamp.RESULTS:The rats in HF group exhibited significantly reduced expression and activity of AMPK/SIRT1 in skeletal muscles(P<0.05).More-over,mitochondrial oxidative stress markers,reactive oxygen species(ROS)and malondialdehyde(MDA),were marked-ly elevated(P<0.05),and the activity of antioxidant enzymes glutathione peroxidase(GPX)and manganese superoxide dismutase(MnSOD)was significantly decreased in HF group(P<0.05).There was also a notable decline in the activity of citrate synthase(P<0.05),a marker of mitochondrial oxidative capacity,and the copy number of mitochondrial DNA in HF group.These changes were correlated with significantly decreased GIR60-120 and DGU(P<0.05).Notably,Met treat-ment(HF+Met)restored the AMPK/SIRT1 expression and activity,improved mitochondrial function,and reduced oxida-tive stress,leading to improved insulin sensitivity(P<0.05).However,these beneficial effects of Met were reversed by the MC4R antagonist SHU9119 in HF+Met+SHU9119 group.CONCLUSION:Treatment with Met enhances skeletal muscle AMPK/SIRT1 expression and activity,reverses mitochondrial dysfunction,and improves insulin resistance in HF-fed rats.These effects might be mediated through the activation of hypothalamic MC4R.

Aim To investigate the relationship between the triglyceride glucose-body mass index(TyG-BMI)and the extent of coronary artery disease in patients with premature coronary artery disease(PCAD).Methods A ret-rospective analysis was performed on the clinical data of 533 PCAD patients diagnosed by coronary angiography at Xuzhou Central Hospital from January 2023 to December 2023.Patients were divided into three groups based on TyG-BMI ter-tiles:group T1(TyG-BMI ≤228.27),group T2(228.27＜TyG-BMI ≤248.90),and group T3(TyG-BMI＞248.90).The extent of coronary artery disease was quantitatively assessed using the Gensini score,which was categorized into low/mid-risk(Gensini score≤70)and high-risk(Gensini score＞70)groups.Univariate and multivariate Logistic regression models were used to analyze the relationship between TyG-BMI and the occurrence of high Gensini score coronary artery dis-ease.Spearman correlation analysis was employed to assess the correlation between TyG-BMI and Gensini score.ROC curve was used to analyze the predictive value of TyG-BMI for severe coronary artery disease.Results Patients in group T3 had more multi-vessel lesions and higher Gensini scores(P＜0.001).Logistic regression analysis showed that a high level of the TyG-BMI was an independent risk factor for severe coronary artery disease in patients with PCAD(OR=1.031,95％CI:1.021～1.040,P＜0.001).Compared with group T1,the risk for severe coronary artery disease in group T3 was 3.57-fold higher(OR=3.57,95％CI:2.170～5.870,P＜0.001).Spearman correlation analysis showed that TyG-BMI was positively correlated with the severity of coronary artery disease(r=0.202,P＜0.001).ROC curve a-nalysis showed that the area under curve of TyG-BMI was 0.704(95％CI:0.656～0.752),which was superior to that of the TyG index in predictive value(P＜0.05).Conclusion A high level of the TyG-BMI can be used as an effective predictor of severe coronary artery disease in patients with PCAD.

Objective:To construct fused cancer cell/neutrophil membrane-coated polydopamine nanoparticles chelated with manganese ions (Ⅱ) (PMNP@FMs) and explore the potential for targeted pancreatic cancer fluorescence imaging and MRI.Methods:Cancer cell membranes fused with neutrophil membranes were encapsulated on the surface of polydopamine nanoparticles chelated with manganese ions (Ⅱ) (PMNPs) to prepare PMNP@FMs. The morphology, structure, and MRI performance of the product were characterized. The cytotoxicity of PMNP@FMs towards human pancreatic cancer cells (PANC-1) and normal human pancreatic ductal epithelial cells (hTERT-HPNE) was evaluated using cell counting kit (CCK)-8, and in vivo toxicity was assessed in healthy mice. PANC-1 pancreatic cancer xenograft nude mouse models were established for in vivo fluorescence imaging and MRI. Data were analyzed using the independent-sample t test, repeated measures analysis of variance and the least significance difference method. Results:PMNP@FMs exhibited a core-shell structure with a diameter of (112.81±8.64) nm, negative surface charge, and good dispersibility. The T 1 relaxivity of PMNPs was 18.81±0.22, which was 4.1 times higher than that of gadopentetate dimeglumine (Gd-DTPA) (4.55±0.24; t=75.54, P<0.001). Co-culture of PMNPs and PMNP@FMs with hTERT-HPNE and PANC-1 cells for 24 h resulted in cell viability above 90% within the concentration range of 0-500 μg/ml. PMNP@FMs did not affect mouse survival and showed no apparent organ damage. In vivo fluorescence imaging and MRI revealed that PMNP@FMs accumulated highly in tumors and reached the peak 24 h post intravenous administration (relative MR signal: 1.35±0.01, fluorescence intensity: (1.20±0.25)×10 10), surpassing the peak observed in the control group (1.22±0.01, (3.87±0.50)×10 9;F values: 11.03-188.01, t values: 18.20, 5.64, all P<0.05), with hepatic metabolism being the primary route of clearance. Conclusion:PMNP@FMs demonstrate a potential for targeted pancreatic cancer fluorescence imaging and MRI, offering promising prospect for precise diagnosis of early-stage pancreatic cancer.

Objective:To investigate the denoising performance of different deep learning (DL) strategies on low-dose brain 18F-FDG PET images. Methods:This retrospective methodological study was conducted on brain PET/CT images of 50 patients (35 males, 15 females, age 20-87 years) who received 3.7MBq/kg 18F-FDG at the Fifth Affiliated Hospital of Sun Yat-sen University between May 2023 and January 2024. Full-dose PET data were acquired with 2min scan. CT scans were acquired before PET scanning. Low-dose PET sinograms were generated by down-sampling the full-dose list mode data to 1/2, 1/4, and 1/20 of full-dose count level. Both full-dose and low-dose sinograms were reconstructed with random, CT-based attenuation and scatter corrections using the three-dimensional (3D) ordered-subsets expectation maximization (OSEM) algorithm (2 iterations, 20 subsets). A total of 4 DL denoising methods were established: (1) 3D conditional generative adversarial networks (GAN) using only low-dose PET as input (GAN-1); (2) 3D attention-based GAN (AttGAN) with low-dose PET input (AttGAN-1); (3) 3D AttGAN with low-dose PET and CT inputs (AttGAN-2); (4) 3D AttGAN with frequency-separation using low-dose PET and CT inputs (AttGAN-FS-2). For AttGAN-FS-2, during the frequency division process, high- and low-frequency components were extracted from the PET reconstructed images via Fourier transform, then inversed Fourier transform, denoised separately, and finally combined to produce the final denoised images. The dataset was separated into training (70%), validation (10%) and testing (20%) sets using simple random sampling without replacement with a fixed random seed. A 5-fold cross-validation scheme was then applied to test all 50 patients. Performance was evaluated against full-dose PET using normalized mean square error (NMSE), structural similarity (SSIM), peak signal-to-noise ratio (PSNR), contrast-to-noise ratio (CNR), SUV mean and SUV max bias of selected brain ROIs. Wilcoxon signed rank test was used to analyze the differences between the denoising methods. Results:AttGAN-FS-2 showed the best performance among all dose levels, with statistical difference as compared by low-dose PET and GAN-1 denoised images for NMSE, SSIM, PSNR, and CNR ( Z values: 2.92-6.15, all P<0.005). NMSE, SSIM quantitative evaluation results (median) of each model at 1/20 dose were: GAN-1: 0.08, 0.87, AttGAN-1: 0.08, 0.88, AttGAN-2: 0.07, 0.89, AttGAN-FS-2: 0.06, 0.91, respectively ( Z values: 3.24-5.77, all P<0.005). Conclusion:The DL-based method combined with multiple strategies AttGAN-FS-2 shows improved denoising performance for low-dose brain PET images.

Objective:To develop and validate machine learning-based radiomics models using preoperative CT images for individualized prediction of mitotic index (MI) in patients with gastrointestinal stromal tumors (GIST).Methods:The study was a case-control study. The data of 348 GIST patients confirmed by pathology were retrospectively collected from two independent medical centers: the Affiliated Hospital of Qingdao University (center 1) and Shandong Provincial Hospital Affiliated to Shandong First Medical University (center 2), covering the period from January 2013 to June 2018. Patients from center 1 were divided into a training cohort (176 cases) and an internal validation cohort (75 cases) at a ratio of 7∶3 using random sampling. Patients from center 2 served as an independent external validation cohort (97 cases). The primary endpoint was MI, categorized into high MI (145 cases) and low MI (203 cases) groups. Radiomic features were extracted from the portal venous phase images of preoperative contrast-enhanced CT scans. Five machine learning algorithms, including logistic regression, support vector machine, random forest, decision tree, and extreme gradient boosting (XGBoost),were employed to construct MI prediction models. The optimal model was identified using receiver operating characteristic curves. An individualized prediction model was developed by integrating the the optimal machine learning model combined with selected independent clinical factors, and the importance of features was visualized using Shapley Additive Explanation (SHAP) analysis. Patients were followed up, and Kaplan-Meier curves along with log-rank tests were used to evaluate recurrence-free survival (RFS) differences between the predicted high MI and low MI groups.Results:Among the five constructed machine learning models, the XGBoost model demonstrated the best predictive performance, with area under the curve (AUC) of 0.809 (95% CI 0.738-0.872), 0.693 (95% CI 0.571-0.809), and 0.718 (95% CI 0.605-0.822) in the training cohort, internal validation cohort, and external validation cohort, respectively. An individualized prediction model combining the XGBoost model with independent clinical factors (tumor location and tumor size) was developed. The model achieved AUC of 0.843 (95% CI 0.785-0.899), 0.791 (95% CI 0.680-0.894), and 0.777 (95% CI 0.678-0.861) in the training cohort, internal validation cohort, and external validation cohort, respectively. SHAP analysis indicated that radiomic features had the highest predictive impact. In both the training cohort and internal validation cohort, the RFS of patients predicted to be in the high MI group was lower than that of the low MI group, with statistically significant differences ( χ2=14.58, 9.52, both P<0.001). However, there was no statistically significant difference in RFS in the external validation set ( χ2=6.18, P=0.080). Conclusions:The optimal XGBoost model based on radiomic features extracted from preoperative portal venous phase CT images, when combined with clinical factors, can effectively predict the MI of GIST patients.

Objective To explore the mechanism of S100A9 derived from non-small cell lung cancer(NSCLC)in regulating invasion,metastasis and activating the brain microvascular endothelium of the metastatic niche.Methods R language was used to extract RNAseq data from the TCGA database and a paired-sample T-test was employed to analyze the expression of S100A9 in NSCLC tissues and normal lung tissues.Visualization was conducted using the ggplot2 package;the proportional hazards assumption test and survival regression were performed using the survival package to compare the prognosis between the high/low expression groups of S100A9,and visualization was carried out using the survminer package and ggplot2 package.RT-qPCR and Western Blot were used to detect the expression differences of S100A9 in NSCLC cell lines(A549,NCI-H1299)and normal lung epithelial cells(BEAS-2B).An in vitro co-culture of A549 cells and human brain microvascular endothelial cells(hCMEC/D3)was established to construct a blood-tumor barrier(BTB)model.Additionally,siS100A9 knock-down A549 cell strains were constructed.Scratch healing and Transwell assays were performed to assess the changes in the migration and invasion abilities of A549 cells in different treatment groups.CCK-8 and flow cytometry were used to examine the proliferative activity and cell cycle effects of HCMEC/D3 cells treated with varying concen-trations of S100A9.RT-qPCR and Western Blot were employed to investigate the expression changes of receptors for advanced glycation endproducts(RAGE),Toll-like receptor 4(TLR4),and tumor transendothelial migration-related adhesion molecules(ICAM-1,VCAM-1,ALCAM)in hCMEC/D3 cells treated with different concen-trations of S100A9.Furthermore,CCK-8,RT-qPCR,and Western Blot were utilized to assess the recovery of proliferative activity and adhesion molecule expression in hCMEC/D3 cells stimulated with different concentrations of S100A9 after pretreatment with FPS-ZM1 and TAK242 to block RAGE and TLR4 pathways,respectively.Results The RNAseq data mining and analysis from the TCGA database revealed that the expression of S100A9 in lung cancer tissue samples was significantly higher than that in normal lung tissue samples(P=0.03).The Kaplan-Meier survival curve graph showed that the survival probability of the S100A9 high-expression group was lower than that of the S100A9 low-expression group,suggesting that the high expression of S100A9 was significantly associated with a poorer overall survival period for patients(HR=1.46(1.10-1.95),P=0.01).In the cell experiments,S100A9 was highly expressed in NSCLC(P<0.05).Knockdown of S100A9 inhibited the migration and invasion of A549 cells(P<0.05).The average migration inhibition rate of the knockdown group at 6,12,24,36,and 48 hours was 80.61%,75.70%,73.78%,69.54%,and 56.96%respectively,and the average invasion inhibition rate was 57.38%(at 48 hours).Meanwhile,the proliferative activity and cell cycle of hCMEC/D3 cells in the BTB model were regulated positively(P<0.05).Mechanistically,S100A9 promoted the crosstalk between A549 and hCMEC/D3 cells through RAGE and TLR4,upregulating the expression of ICAM-1,VCAM-1,and ALCAM in hCMEC/D3 cells(P<0.05).Recovery experiments confirmed that the S100A9-RAGE/TLR4 regulatory axis could affect the endothelial adhesion process during lung cancer brain metastasis(P<0.05).Conclusion The S100A9-RAGE/TLR4 axis is associated with the progression of lung cancer brain metastasis.Knockdown of S100A9 can inhibit the invasion and metastasis of lung cancer cells.Blocking downstream RAGE and TLR4 receptors can attenuate the proliferative growth of brain microvascular endothelium and inhibit the formation of a pre-metastatic adhesive microenvironment between lung cancer cells and brain microvascular endothelium.

AIM:To explore the central mechanisms by which metformin(Met)attenuates insulin resistance in high-fat diet(HF)-fed rats.METHODS:Forty healthy male Sprague-Dawley rats were randomly divided into 4 groups:normal chow(NC)group,HF group,HF+Met group,and HF+Met+SHU9119[melanocortin 4 receptor(MC4R)antagonist]group,with 10 rats per group.Treatments with HF and Met lasted for 12 weeks,while SHU9119 was injected for the last 10 d.Skeletal muscle AMP-activated protein kinase(AMPK)and silent information regulator 1(SIRT1)ex-pression and activity were measured,along with mitochondria oxidative stress markers,mitochondrial function and quanti-ty.Systemic and skeletal muscle insulin sensitivity were assessed using the average glucose infusion rate from 60 to 120 min(GIR60-120)and 2-deoxyglucose uptake(DGU)during hyperinsulinemic-euglycemic clamp.RESULTS:The rats in HF group exhibited significantly reduced expression and activity of AMPK/SIRT1 in skeletal muscles(P<0.05).More-over,mitochondrial oxidative stress markers,reactive oxygen species(ROS)and malondialdehyde(MDA),were marked-ly elevated(P<0.05),and the activity of antioxidant enzymes glutathione peroxidase(GPX)and manganese superoxide dismutase(MnSOD)was significantly decreased in HF group(P<0.05).There was also a notable decline in the activity of citrate synthase(P<0.05),a marker of mitochondrial oxidative capacity,and the copy number of mitochondrial DNA in HF group.These changes were correlated with significantly decreased GIR60-120 and DGU(P<0.05).Notably,Met treat-ment(HF+Met)restored the AMPK/SIRT1 expression and activity,improved mitochondrial function,and reduced oxida-tive stress,leading to improved insulin sensitivity(P<0.05).However,these beneficial effects of Met were reversed by the MC4R antagonist SHU9119 in HF+Met+SHU9119 group.CONCLUSION:Treatment with Met enhances skeletal muscle AMPK/SIRT1 expression and activity,reverses mitochondrial dysfunction,and improves insulin resistance in HF-fed rats.These effects might be mediated through the activation of hypothalamic MC4R.

Objectives:To investigate the effect of circular RNA (circRNA) hsa_circ_0019217 on the biological function of chorionic trophoblast cell line (HTR8/SVneo) and its mechanism in the occurrence of pre-eclampsia (PE).Methods:The circRNA expression database was used to analyze the differentially expressed circRNA in placental tissues of PE women. The expression levels of hsa_circ_0019217 and miR-526b-5p were detected by reverse transcription real-time fluorescent quantitative PCR (RT-qPCR), and the subcellular localization of hsa_circ_0019217 was detected by fluorescence in situ hybridization. The proliferation, migration and invasion of trophoblast cells were detected by cell counting kit-8 (CCK-8) assay and transwell assay. Western blot and enzyme-linked immunosorbent assay (ELISA) were used to verify the effects of hsa_circ_0019217, miR-526b-5p and decorin (DCN) on matrix metalloproteinase 2 (MMP2), tissue inhibitor of metalloproteinase 2 (TIMP2), placental growth factor (PlGF) and human chorionic gonadotropin (hCG) protein expression levels. Dual luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay were used to verify the interaction between hsa_circ_0019217, miR-526-5p and DCN.Results:(1) The analysis of circRNA expression database showed that the expression level of hsa_circ_0019217 was significantly increased in the placental tissues of PE women (fold change=67, P<0.05), and it was mainly located in the cytoplasm. (2) Knockdown of hsa_circ_0019217 promoted the proliferation, migration and invasion of HTR8/SVneo cells, increased the expression levels of MMP2 and PlGF, and decreased the expression levels of TIMP2 and hCG in HTR8/SVneo cells. (3) Hsa_circ_0019217 targeted adsorption of miR-526b-5p, and inhibition of miR-526b-5p reduced the proliferation, migration and invasion of HTR8/SVneo cells. The expression levels of MMP2 and PlGF in HTR8/SVneo cells were increased, and the expression levels of TIMP2 and hCG were decreased. Hsa_circ_0019217 knockdown and inhibiting miR-526b-5p could reverse the effect of hsa_circ_0019217 knockdown on promoting the proliferation, migration and invasion of HTR8/SVneo cells, and reversed the effect of hsa_circ_0019217 knockdown on the protein expression levels of MMP2, PlGF, TIMP2 and hCG. (4) miR-526b-5p targeted DCN in HTR8/SVneo cells, hsa_circ_0019217 knockdown reduced the expression level of DCN, and inhibiting miR-526b-5p increased the expression level of DCN. When hsa_circ_0019217 and miR-526b-5p were inhibited simultaneously, there was no significant change in the protein expression level of DCN. (5) Overexpression of miR-526b-5p promoted the proliferation, migration and invasion of HTR8/SVneo cells, while overexpression of DCN inhibited the proliferation, migration and invasion of HTR8/SVneo cells. Simultaneous overexpression of miR-526b-5p and DCN reversed the effects of miR-526b-5p overexpression on cell proliferation, migration and invasion. Overexpression of miR-526b-5p increased the expression levels of MMP2 and PlGF and decreased the expression levels of TIMP2 and hCG in HTR8/SVneo cells. Overexpression of DCN reduced the expression levels of MMP2 and PlGF and increased the expression levels of TIMP2 and hCG. Overexpression of miR-526b-5p and DCN reversed the effects of miR-526b-5p on the expression of these proteins. Conclusion:Hsa_circ_0019217 regulates the expression of DCN by adsorption of miR-526b-5p, thereby affecting the proliferation, migration and invasion of trophoblast cells and regulating the protein expression levels of MMP2, TIMP2, PlGF and hCG, which might be used as a target for early prevention of PE.