Background::The ratio of high-density lipoprotein cholesterol (HDL-C) to low-density lipoprotein cholesterol (LDL-C) predicts cardiovascular disease (CVD) endpoints, yet its prognostic validity in high-risk populations and for type 2 diabetes mellitus (T2DM)-related adverse events remains unestablished.Methods::This study included 32,609 people aged 35-75 years in Fujian Province, China, who were at high risk for CVD. The primary endpoint was all-cause mortality during follow-up. Cox proportional hazard models and restricted cubic spline (RCS) analysis were used to evaluate the correlation between the HDL-C/LDL-C ratio and the endpoints.Result::On the basis of the restricted RCS curve, the participants were classified as having a low (< 0.3), middle (0.3-0.5), or high (> 0.5) HDL-C/LDL-C ratio. Multivariate Cox regression analyses revealed that the risk of all-cause mortality (HR = 1.48, 95% CI 1.14-1.93, p < 0.01 for low; HR = 1.30, 95% CI 1.06-1.58, p < 0.05 for high) was increased in the low and high groups. Participants without T2DM who were at high risk for CVD had similar prognoses (HR = 1.65, 95% CI 1.19-2.28, p < 0.01 for low; HR = 1.35, 95% CI 1.05-1.74, p < 0.01 for high). However, this association was not found in participants with T2DM who were at high risk for CVD. Conclusion::HDL-C/LDL-C can be used to predict the prognosis of individuals at high risk for CVD, and maintaining HDL-C/LDL-C ratios between 0.3 and 0.5 may be the most helpful range for this population. Furthermore, maintaining this ratio range holds clinical significance for cohorts without T2DM, although further exploration is needed in this T2DM cohort.

Background::The ratio of high-density lipoprotein cholesterol (HDL-C) to low-density lipoprotein cholesterol (LDL-C) predicts cardiovascular disease (CVD) endpoints, yet its prognostic validity in high-risk populations and for type 2 diabetes mellitus (T2DM)-related adverse events remains unestablished.Methods::This study included 32,609 people aged 35-75 years in Fujian Province, China, who were at high risk for CVD. The primary endpoint was all-cause mortality during follow-up. Cox proportional hazard models and restricted cubic spline (RCS) analysis were used to evaluate the correlation between the HDL-C/LDL-C ratio and the endpoints.Result::On the basis of the restricted RCS curve, the participants were classified as having a low (< 0.3), middle (0.3-0.5), or high (> 0.5) HDL-C/LDL-C ratio. Multivariate Cox regression analyses revealed that the risk of all-cause mortality (HR = 1.48, 95% CI 1.14-1.93, p < 0.01 for low; HR = 1.30, 95% CI 1.06-1.58, p < 0.05 for high) was increased in the low and high groups. Participants without T2DM who were at high risk for CVD had similar prognoses (HR = 1.65, 95% CI 1.19-2.28, p < 0.01 for low; HR = 1.35, 95% CI 1.05-1.74, p < 0.01 for high). However, this association was not found in participants with T2DM who were at high risk for CVD. Conclusion::HDL-C/LDL-C can be used to predict the prognosis of individuals at high risk for CVD, and maintaining HDL-C/LDL-C ratios between 0.3 and 0.5 may be the most helpful range for this population. Furthermore, maintaining this ratio range holds clinical significance for cohorts without T2DM, although further exploration is needed in this T2DM cohort.

A VLi-net based cell nucleus segmentation method integrating convolutional neural networks(CNN)and Vision Transformer(ViT)is proposed to address the limitation that the U-Net with CNN as its backbone is only proficient in capturing local features and has a restricted receptive field.Firstly,to mitigate challenges such as high cost of data annotation and insufficient annotated data,text annotations are introduced to enhance the network's understanding of image information.Secondly,to improve the segmentation performance of VLi-net,ViT and CNN are combined to fully extract global and local features,with multi-receptive field convolution features incorporating into the ViT structure for effectively mitigating the issues of limited local information interaction and single feature representation in ViT.Finally,an interactive fusion module(ViFusion)is used to efficiently fuse the multi-level features from the CNN and ViT branches.Experimental results show that VLi-net achieves a Dice coefficient of 80.85%and a mean intersection over union(MIoU)of 66.83%on the MoNuSeg dataset,obtains a Dice coefficient of 80.53%and a MIoU of 67.54%on the DSB-2018 dataset,and has a Dice coefficient of 86.87%and a MIoU of 77.44%on the TNBC dataset.These findings confirm that VLi-net outperforms other methods across multiple experimental metrics.

Objective:To investigate the drug resistance gene characteristics, plasmid characteristics and genome tracing of Shigella sonnei causing a bacillary dysentery outbreak in Shandong Province. Methods:Sixty-five Shigella sonnei strains isolated from a 2021 outbreak in a county of Shandong Province were analyzed using antimicrobial susceptibility testing, whole genome sequencing (WGS), characterization of resistance and virulence genes, plasmid profiling, core genome multilocus sequence typing (cgMLST), and single nucleotide polymorphism (SNP) analysis. Results:All isolates had the same resistance phenotype and genotypes and were multidrug-resistant ESBL-producing Shigella sonnei, carrying important virulence genes. Plasmid analysis revealed a conserved genetic arrangement, pil( M/ N/ O2/ P)-tra( F/ H/ J/ K/ N/ O/ P/ Q)-IS Ecp1- blaCTX-M-14-Tn 903- yub( J/ I/ F/ G/ E/ D), and shared across strains from diverse regions and bacterial species. The cgMLST and SNP analyses demonstrated concordant clustering, with all 65 outbreak-related strains forming a single cluster alongside human-derived strains from Guangxi. Conclusion:The ESBL-producing Shigella sonnei responsible for the outbreak shares a homologous relationship with Guangxi human-derived strains, and the detected resistance plasmids and virulence genes underscore the need to strengthen drug resistance surveillance and genome tracing.

AIM:This study aims to investigate whether vitamin E succinate(VES)induces autophagy in hu-man gastric cancer cells through the promotion of mitochondria-associated endoplasmic reticulum membranes(MAMs).METHODS:Human gastric cancer cell lines MKN28 and MKN45 were cultured in vitro.Cell viability was assessed us-ing the CCK8 assay,and two cell growth curves were plotted to determine the treatment concentration of VES.Control groups,VES dose groups(MKN28:5,10,20,and 40 mg/L;MKN45:10,20,40,and 80 mg/L),an autophagy-posi-tive control group(rapamycin,RAPA,100 nmol/L),and a MAMs-positive control group(oligomycin A,10 mg/L)were set up.Cells were harvested after 24 h of treatment for subsequent experiments.The formation of autophagosomes and MAMs was observed using transmission electron microscopy.The expression levels of autophagy-related proteins,includ-ing beclin-1,LC3-II/LC3-I,and p62,were detected by Western blot.MAMs labeled with split green fluorescent protein(GFP)were visualized by fluorescence microscopy.The expression of mitofusin 2(MFN2),a key molecule of MAMs,was also detected by Western blot.To inhibit MFN2 specifically,the cells were treated with mitochondrial fusion inhibitor 8(MFI8)and simultaneously transfected with an MFN2 plasmid to achieve MFN2 overexpression(OE-MFN2).The cells were divided into control group,MFI8(20 μmol/L)group,VES groups(20 mg/L for MKN28 cells and 40 mg/L for MKN45 cells),VES+MFI8 group,OE-MFN2+MFI8 group and OE-MFN2+VES+MFI8 group.The MAMs were visualized by fluorescence microscopy,and the expression changes of MFN2,beclin-1 and LC3-II/I were detected by Western blot.RESULTS:The results of the CCK8 assay showed that VES significantly inhibited the viability of both human gastric can-cer cell lines(P<0.05).After VES treatment,the formation of typical autophagosomes and MAMs was observed in both cell lines by transmission electron microscopy.Fluorescence microscopy showed a significant increase in GFP signals of MAMs.Western blot analysis showed that with increasing doses of VES,the expression levels of MFN2,beclin-1,and LC3-II/I were significantly up-regulated,while that of p62 was significantly down-regulated(P<0.05).Compared with VES group,the cells pretreated with MFI8 followed by VES exposure showed markedly reduced GFP signals of MAMs and much lower protein levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).Transfection with an MFN2 overexpression plasmid rescued MFN2 expression.Compared with VES+MFI8 group,the cells in OE-MFN2+VES+MFI8 group had much higher protein expression levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).CONCLUSION:The VES may partici-pates in the regulation of autophagy in human gastric cancer cells by promoting the formation of MAMs.

Objective To propose a classification model based on a pseudo-bag strategy and feature adjustment for whole slide imaging in pathology.Methods A pseudo-bag generator was constructed to divide a parent bag into 3 pseudo-bags for increasing the number of training bags.Then,a pseudo-bag learning method based on Nystr?m-based algorithm for approximating self-attention and a selective feature fusion method were employed to process the pseudo-bags.Specifically,the pseudo-bag learning method based on Nystr?m-based algorithm for approximating self-attention reduced computational complexity through an improved multi-head self-attention mechanism while deeply extracting instance features to obtain pseudo-bag classification predictions,thereby enhancing pseudo-bag classification accuracy;and the selective feature fusion method refined pseudo-bag features by filtering and extracting relevant instances.Finally,the model adjusted bag features by extracting confounding factors to avoid interference from irrelevant information and further improve classification accuracy.Results The proposed model was evaluated on two datasets(CAMELYON-16 and TCGA-NSCLC)and compared with 10 other methods,and the results demonstrated that the proposed model achieved the best performance.The proposed method reached an accuracy of 0.943 on the CAMELYON-16 dataset and 0.906 on the TCGA-NSCLC dataset.Conclusion The proposed model can significantly improve the accuracy of whole-slide pathological image classification by effectively mitigating the overfitting and avoiding interference from irrelevant information.

This study aims to develop a fully automated method for tooth segmentation and root canal measurement based on cone beam computed tomography (CBCT) images, providing objective, efficient, and accurate measurement results to guide and assist clinicians in root canal diagnosis grading, instrument selection, and preoperative planning. The method utilized Attention U-Net to recognize tooth descriptors, cropped regions of interest (ROIs) based on the center of mass of these descriptors, and applied an integrated deep learning method for segmentation. The segmentation results were mapped back to the original coordinates and position-corrected, followed by automatic measurement and visualization of root canal lengths and angles. The results indicated that the Dice coefficient for segmentation was 96.42%, the Jaccard coefficient was 93.11%, the Hausdorff Distance was 2.07 mm, and the average surface distance was 0.23 mm, all of which surpassed existing methods. The relative error of the root canal working length measurement was 3.15% (< 5%), the curvature angle error was 2.85 °, and the correct classification rate of the treatment difficulty coefficient was 90.48%. The proposed methods all achieved favorable results, which can provide an important reference for clinical application.
Cone-Beam Computed Tomography/methods* ; Deep Learning ; Humans ; Dental Pulp Cavity/diagnostic imaging* ; Image Processing, Computer-Assisted/methods*

AIM:This study aims to investigate whether vitamin E succinate(VES)induces autophagy in hu-man gastric cancer cells through the promotion of mitochondria-associated endoplasmic reticulum membranes(MAMs).METHODS:Human gastric cancer cell lines MKN28 and MKN45 were cultured in vitro.Cell viability was assessed us-ing the CCK8 assay,and two cell growth curves were plotted to determine the treatment concentration of VES.Control groups,VES dose groups(MKN28:5,10,20,and 40 mg/L;MKN45:10,20,40,and 80 mg/L),an autophagy-posi-tive control group(rapamycin,RAPA,100 nmol/L),and a MAMs-positive control group(oligomycin A,10 mg/L)were set up.Cells were harvested after 24 h of treatment for subsequent experiments.The formation of autophagosomes and MAMs was observed using transmission electron microscopy.The expression levels of autophagy-related proteins,includ-ing beclin-1,LC3-II/LC3-I,and p62,were detected by Western blot.MAMs labeled with split green fluorescent protein(GFP)were visualized by fluorescence microscopy.The expression of mitofusin 2(MFN2),a key molecule of MAMs,was also detected by Western blot.To inhibit MFN2 specifically,the cells were treated with mitochondrial fusion inhibitor 8(MFI8)and simultaneously transfected with an MFN2 plasmid to achieve MFN2 overexpression(OE-MFN2).The cells were divided into control group,MFI8(20 μmol/L)group,VES groups(20 mg/L for MKN28 cells and 40 mg/L for MKN45 cells),VES+MFI8 group,OE-MFN2+MFI8 group and OE-MFN2+VES+MFI8 group.The MAMs were visualized by fluorescence microscopy,and the expression changes of MFN2,beclin-1 and LC3-II/I were detected by Western blot.RESULTS:The results of the CCK8 assay showed that VES significantly inhibited the viability of both human gastric can-cer cell lines(P<0.05).After VES treatment,the formation of typical autophagosomes and MAMs was observed in both cell lines by transmission electron microscopy.Fluorescence microscopy showed a significant increase in GFP signals of MAMs.Western blot analysis showed that with increasing doses of VES,the expression levels of MFN2,beclin-1,and LC3-II/I were significantly up-regulated,while that of p62 was significantly down-regulated(P<0.05).Compared with VES group,the cells pretreated with MFI8 followed by VES exposure showed markedly reduced GFP signals of MAMs and much lower protein levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).Transfection with an MFN2 overexpression plasmid rescued MFN2 expression.Compared with VES+MFI8 group,the cells in OE-MFN2+VES+MFI8 group had much higher protein expression levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).CONCLUSION:The VES may partici-pates in the regulation of autophagy in human gastric cancer cells by promoting the formation of MAMs.

Objective To propose a classification model based on a pseudo-bag strategy and feature adjustment for whole slide imaging in pathology.Methods A pseudo-bag generator was constructed to divide a parent bag into 3 pseudo-bags for increasing the number of training bags.Then,a pseudo-bag learning method based on Nystr?m-based algorithm for approximating self-attention and a selective feature fusion method were employed to process the pseudo-bags.Specifically,the pseudo-bag learning method based on Nystr?m-based algorithm for approximating self-attention reduced computational complexity through an improved multi-head self-attention mechanism while deeply extracting instance features to obtain pseudo-bag classification predictions,thereby enhancing pseudo-bag classification accuracy;and the selective feature fusion method refined pseudo-bag features by filtering and extracting relevant instances.Finally,the model adjusted bag features by extracting confounding factors to avoid interference from irrelevant information and further improve classification accuracy.Results The proposed model was evaluated on two datasets(CAMELYON-16 and TCGA-NSCLC)and compared with 10 other methods,and the results demonstrated that the proposed model achieved the best performance.The proposed method reached an accuracy of 0.943 on the CAMELYON-16 dataset and 0.906 on the TCGA-NSCLC dataset.Conclusion The proposed model can significantly improve the accuracy of whole-slide pathological image classification by effectively mitigating the overfitting and avoiding interference from irrelevant information.

ObjectiveThis study aimed to investigate the intervention effect of Renqing Mangjue on the malignant transformation of gastric mucosal epithelial cells induced by N-methyl-N′-nitro-N-nitrosoguanidine （MNNG） and to explore its molecular mechanism in preventing precancerous lesions of gastric cancer based on the cyclic guanosine monophosphate （cGMP）/protein kinase G （PKG）/mitogen-activated protein kinase （MEK）/extracellular signal-regulated kinase （ERK） signaling pathway. MethodsHuman gastric mucosal epithelial cells （GES-1） were initially induced by MNNG to establish a precancerous cell model （MC cells）. The effective concentration of MNNG for inducing malignant transformation in GES-1 cells was screened using the cell proliferation activity decection （CCK-8） assay， and the effective concentration of Renqing Mangjue for inhibiting the proliferation of transformed GES-1 cells was also determined. GES-1 cells were divided into a blank control group， a model group， and treatment groups with Renqing Mangjue at concentrations of 1， 3， 10， and 30 mg·L^-1. Furthermore， the effects of Renqing Mangjue on the migratory ability and epithelial-mesenchymal transition （EMT） characteristics of GES-1 malignant transformed cells were evaluated using Transwell migration assays， wound healing assays， and real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR）. Additionally， candidate chemical components and target sites of Renqing Mangjue were obtained from the TCMIP v2.0 database， and disease targets at various stages of gastric cancer precursors were sourced from the Gene Expression Omnibus （GEO） database. Pathway enrichment analysis was performed using the Metascape database to predict the potential mechanisms of action of Renqing Mangjue. Finally， the protective mechanism of Renqing Mangjue against gastric cancer precursors was validated through Western blot analysis. ResultsAt a concentration of 20 μmol·L^-1， MNNG exhibited an inhibition rate of approximately 50% on GES-1 cells （P<0.01）， and at this concentration， the GES-1 cells displayed biological characteristics indicative of malignant transformation. In contrast， Renqing Mangjue had no significant effect on the proliferation of normal GES-1 cells， but significantly inhibited the proliferation of MC cells （P<0.01） and markedly reduced their migratory capacity （P<0.01）. Moreover， it also increased the mRNA expression level of E-cadherin during the EMT process （P<0.05）， while inhibiting the expression of both N-cadherin and the transcription factor Snail mRNA （P<0.05， P<0.01）. Network predictions suggested that Renqing Mangjue may prevent gastric cancer precursors through modulating the cGMP/PKG and MAPK/ERK signaling pathways. Furthermore， Western blot results indicated that Renqing Mangjue upregulated the expression of PKG and NPRB （B-type natriuretic peptide receptor） proteins in the cGMP/PKG pathway （P<0.01）， while downregulating the expression of the downstream proteins MEK and ERK （P<0.05， P<0.01）. ConclusionIn summary， Renqing Mangjue can prevent gastric cancer precursors by inhibiting the proliferation and migration of malignant transformed GES-1 cells， thereby delaying the EMT process. The underlying mechanisms may be related to the activation of the cGMP/PKG pathway and the inhibition of the MEK/ERK signaling pathway.