Objective To identify the enhancer profile marked by histone H3K27ac modification in high-grade intraepithelial neoplasia(HGIN)in order to reveal the novel regulatory mechanism of HGIN pathogensis.Methods Gastric tissue samples were collected from Department of Gastroenterology of Army Medical Center of PLA between June 2022 and June 2023,including 14 normal gastric tissues(Nor group),31 HGIN tissues(HGIN group)and 17 gastric cancer tissues(GC group).Cleavage under targets and tagmentation(CUT&Tag)technique was employed to capture enhancer regions modified by histone H3K27ac.Multi-omics analysis was performed to identify HGIN-specific active enhancers and their potentially regulated genes.Immunohistochemical profiling was performed to assess differential expression of the gene of interest across clinically stratified specimens,combined with CRISPR-dCas9-mediated ablation of active enhancers to monitor the gene of interest transcriptional dynamics and validate enhancer-mediated regulatory mechanisms.Results Epigenomic sequencing obtained the data with excellent quality,and indicated that obvious remodeling was observed in H3K27ac enhancers in HGIN and GC groups(P<0.05),though no significant difference in the genome-wide distribution of H3K27ac modification among the 3 groups.Combining transcriptome data revealed that enhancer remodeling may up-regulate the expression of the proliferation-related target gene,CD24,in the HGIN tissue;while,inhibiting enhancer activity can notably reduce CD24 expression level(P<0.05).Immunohistochemical assay displayed a positive correlation between the expression levels of CD24 and Ki-67(P<0.001).Conclusion The remodeling of H3K27ac enhancer represents a significant epigenetic feature of the transformation from normal condition to HGIN.Remodeling of H3K27ac enhancer up-regulates CD24,which may facilitate the abnormal proliferation of gastric epithelial cells.

Objective To analyze clinical characteristics of mesenchymal-subtype gastric cancer(Mes-GC)by integrating multi-omics data and explore the characteristics of tumor cells and microenvironment associated with the risk for recurrence.Methods Gastric tumor tissue samples were collected from the patients who visited Department of Gastroenterology of Army Medical Center of PLA from January 2022 to December 2023.Transcriptome and genome sequencing were applied for these tissue samples,including 19 cases of diffuse-type gastric cancer,22 cases of intestinal-type gastric cancer,and 23 cases of mixed-type gastric cancer patients.Bioinformatics analysis was employed to investigate the differences in clinical characteristics and tumor microenvironment between Mes-GC and non-mesenchymal-subtype gastric cancer(non-Mes-GC)by integrating data resources including The Cancer Genome Atlas(TCGA),Gene Expression Omnibus(GEO),and National Genomics Data Center(NGDC).Results Compared to non-Mes-GC patients,Mes-GC ones were characterized by later clinical stages,deeper tumor infiltration,and higher rates of lymph node metastasis.Kaplan-Meier survival analysis confirmed that Mes-GC patients were associated with shorter survival time,poor prognosis as well as increased risk of cancer recurrence(P<0.05).Single-cell RNA sequencing data revealed that tumor cells in Mes-GC showed higher expression levels of the genes related to stemness,metastasis(P<0.05),and epithelial-mesenchymal transition(EMT).And in the tumor microenvironment,there were significant more myeloid cells,smooth muscle cells,endothelial cells and fibroblasts,with the most pronounced elevation in the proportion of fibroblasts(P<0.05).Moreover,the patients with larger proportion of fibroblasts were associated with poorer prognosis.Conclusion Mes-GC tumor cells exhibit higher stemness and EMT characteristics,and stromal cells such as myeloid cells,endothelial cells,and fibroblasts are enriched in the tumor microenvironment.These features may be key factors contributing to poor prognosis and high recurrence rate of Mes-GC.

Objective To identify the enhancer landscape marked by histone H3K27ac modifications in diffuse-type gastric cancer(DGC)tissues,and to elucidate the epigenetic remodeling mechanisms by which active enhancers regulate cachexia-related genes.Methods Gastric mucosal tissue samples were collected from Department of Gastroenterology of Army Medical Center of PLA during January 2022 to March 2023,including 10 normal gastric mucosa tissues(Normal group),10 DGC tissues diagnosed with cachexia(DGC group),and 10 organoids derived from DGC tissues(Organoid group).Using H3K27ac chromatin targeting cleavage and tagmentation(CUT&Tag)technology,genomic modification regions were captured to screen specific active enhancers and their potential target genes in DGC tissues.CRISPR-dCas9 gene editing technology was used to intervene with the enhancers,and the expression of target genes was detected with Western blotting and qRT-PCR.Sixteen female SPF-grade BALB/c Nude mice(6～8 weeks old,weighing 18～21 g)were utilized to establish an orthotopic xenograft tumor model using the human diffuse-type gastric cancer cell line MKN45.Cachexia-related phenotypes were evaluated in 3 groups:normal group(n=4),silencing group(n=6),and control group(n=6).Results Significant differential enhancer regions were identified between DGC and normal gastric mucosa tissues.DGC tissues exhibited a marked increase in enhancer abundance(P<0.05)and signal intensity when compared with the normal counterparts.Integrated analysis of transcriptome data revealed that some of these active enhancers up-regulated the expression of GDF15,a cachexia-associated target gene in DGC.Targeted silencing of the active enhancer of GDF15 using CRISPR/dCas9-KRAB plasmid technology resulted in a significant reduction in GDF15 expression at both mRNA levels(P<0.05)and protein.Results from orthotopic transplantation experiments of DGC demonstrated that silencing of active enhancers alleviated the cachexia phenotype in nude mice(P<0.05).Conclusion DGC exhibits enhancer remodeling,which regulates the expression of the cachexia-associated gene GDF15,and thereby contributes to the pathogenesis and progression of cancer cachexia.

Sustained inflammatory responses are closely related to various severe diseases, and inhibiting the excessive activation of inflammasomes and pyroptosis has significant implications for clinical treatment. Natural products have garnered considerable concern for the treatment of inflammation. Huanglian-Wumei decoction (HLWMD) is a classic prescription used for treating inflammatory diseases, but the necessity of their combination and the exact underlying anti-inflammatory mechanism have not yet been elucidated. Inspired by the supramolecular self-assembly strategy and natural drug compatibility theory, we successfully obtained berberine (BBR)-chlorogenic acid (CGA) supramolecular (BCS), which is an herbal pair from HLWMD. Using a series of characterization methods, we confirmed the self-assembly mechanism of BCS. BBR and CGA were self-assembled and stacked into amphiphilic spherical supramolecules in a 2:1 molar ratio, driven by electrostatic interactions, hydrophobic interactions, and π-π stacking; the hydrophilic fragments of CGA were outside, and the hydrophobic fragments of BBR were inside. This stacking pattern significantly improved the anti-inflammatory performance of BCS compared with that of single free molecules. Compared with free molecules, BCS significantly attenuated the release of multiple inflammatory mediators and lipopolysaccharide (LPS)-induced pyroptosis. Its anti-inflammatory mechanism is closely related to the inhibition of intracellular nuclear factor-kappaB (NF-κB) p65 phosphorylation and the noncanonical pyroptosis signalling pathway mediated by caspase-11.

Objective: To evaluate the effect of exercise prescription intervention among patients with type 2 diabetes mellitus (T2DM), so as to provide the evidence for guiding appropriate physical activity and glycemic control in this population. Methods: In July 2023, T2DM patients managed by two community health service centers in Qingjiangpu District, Huai'an City, Jiangsu Province, were selected as the study participants and randomly assigned divided into a control group and an intervention group. The control group received routine chronic disease management under the basic public health services, while the intervention group, in addition to receiving the same routine chronic disease management, was provided with exercise prescription to guide their physical activity at baseline (T0), after 3 months of intervention (T1), and after 6 months of intervention (T2). Data on weight-related indicators, glycated hemoglobin (HbA1c), and blood lipid were collected through physical examinations and laboratory tests at T0 and after 12 months of intervention (T3). The differences in indicators between the two groups before and after the intervention were analyzed using generalized estimating equations. Results: The intervention group consisted of 197 patients, including 99 males, accounting for 50.25%. The median disease duration was 7.10 (interquartile range, 7.80) years, and 113 patients had suboptimal HbA1c levels, accounting for 57.36%. The control group included 196 patients, including 99 females, accounting for 50.51%. The median disease duration was 6.10 (interquartile range, 7.00) years, and 100 patients had suboptimal HbA1c levels, accounting for 51.02%. Before the intervention, no statistically significant differences were observed between the two groups in gender, educational level, disease duration, pharmacological treatment, smoking, alcohol consumption, and HbA1c levels (all P>0.05). In the intervention group, the proportion of participants engaging in aerobic exercise and strength training increased from 78.17% and 8.12% at T0 to 85.79% and 16.24% at T3, respectively (both P<0.05). The results of the generalized estimating equations revealed significant interactions between group and time for waist-to-hip ratio, total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) following the intervention (all P<0.05). A statistically significant difference in waist-to-hip ratio was found between the two groups (P<0.05), with a greater reduction observed in the intervention group compared to the control group. Significant differences in TC and LDL-C levels were noted across different intervention time points (both P<0.05). Specifically, the intervention group demonstrated reductions of 0.35 mmol/L in TC and 0.42 mmol/L in LDL-C from baseline to follow-up (both P<0.05). Conclusion The 12-month exercise prescription intervention can effectively enhance exercise participation and reduce waist-to-hip ratio, TC, and LDL-C levels among patients with T2DM.

Objective To screen the anti-Streptococcus pneumoniae(Spn)active ingredients in vitro from different po-lar parts of Pilea peltata,and to examine the combined antibacterial effect of the active ingredient and amoxicillin(AMX).Methods A 96-well plate microdilution method was used to determine the minimum inhibitory concentration(MIC)of different polar parts;the most active polar part was separated by preparative high-performance liquid chromatography,and the active ingre-dients were identified using spectral technology.The fractional inhibitory concentration(FIC)of active ingredients and AMX was determined by the 96-well plate chessboard microdilution method.The crystal violet method was used to investigate the effect of ac-tive ingredients on Spn biofilm.The effect of active ingredients on the appearance and morphology of Spn was investigated under the electron microscope.Results The MICs of the petroleum ether part,chloroform part,ethyl acetate part,n-butanol part,and water part were 1.000,1.000,0.500,1.000,and 2.000 mg·mL-1 respectively,among which the ethyl acetate part had the stron-gest antibacterial activity.Three compounds were isolated from ethyl acetate,namely 5,7,3',4'-tetramethoxyflavone 1,8-O-(p-coumaroyl)-1(10)E,4(5)E-humuladien-8-ol 2 and 1-O-p-coumaroyl copaborneol 3.These compounds were all isolated for the first time from Pilea peltate,their MICs against Spn were 200.000,50.000,and 25.000 μg·mL-1 respectively,and the compound 3 had the strongest antibacterial activity;the FIC value of AMX and compound 3 was 0.50,which had a synergistic antibacterial effect on Spn.Both AMX and compound 3 had inhibitory effects on Spn biofilm,but the biofilm inhibition rate of compound 3(59.10±1.04％)was significantly lower than AMX(87.38±0.84)％(P＜0.01);Moreover,there was no significant difference in biofilm inhibition rate between the combination of the two and AMX(P＞0.05).The scanning electron microscope results showed that the bacterial cells in the compound 3 group had a smooth surface but varying degrees of depression.The surface of the bacteri-al cells in the AMX group and the AMX combined compound 3 group showed severe swelling and rupture.Conclusions Fla-vonoids and sesquiterpenoids are both the anti-Spn active components of Pilea peltate.Among them,sesquiterpenoids have more potent antibacterial activity,and their antibacterial action mechanism is related to inhibiting bacterial biofilms.Compound 3 and AMX have a synergistic antibacterial effect on Spn,but its mechanism of action is not by enhancing biofilm inhibition;although compound 3 cannot destroy the cell wall of Spn,it still has a negative impact on the appearance of the bacteria.

Sustained inflammatory responses are closely related to various severe diseases,and inhibiting the excessive activation of inflammasomes and pyroptosis has significant implications for clinical treatment.Natural products have garnered considerable concern for the treatment of inflammation.Huanglian-Wumei decoction(HLWMD)is a classic prescription used for treating inflammatory diseases,but the necessity of their combination and the exact underlying anti-inflammatory mechanism have not yet been elucidated.Inspired by the supramolecular self-assembly strategy and natural drug compatibility theory,we successfully obtained berberine(BBR)-chlorogenic acid(CGA)supramolecular(BCS),which is an herbal pair from HLWMD.Using a series of characterization methods,we confirmed the self-assembly mechanism of BCS.BBR and CGA were self-assembled and stacked into amphiphilic spherical supra-molecules in a 2:1 molar ratio,driven by electrostatic interactions,hydrophobic interactions,and π-πstacking;the hydrophilic fragments of CGA were outside,and the hydrophobic fragments of BBR were inside.This stacking pattern significantly improved the anti-inflammatory performance of BCS compared with that of single free molecules.Compared with free molecules,BCS significantly attenuated the release of multiple inflammatory mediators and lipopolysaccharide(LPS)-induced pyroptosis.Its anti-inflammatory mechanism is closely related to the inhibition of intracellular nuclear factor-kappaB(NF-κB)p65 phosphorylation and the noncanonical pyroptosis signalling pathway mediated by caspase-11.

Objective:To evaluate the clinical utility of dual-layer detector CT integrated with virtual monoenergetic imaging (VMI) and an orthopedic metal artifact reduction (O-MAR) algorithm in improving the image quality of patients after lumbar pedicle screw fixation surgery, and to analyze its impact on different types of artifacts and image quality of different tissues.Methods:The study was a prospective study, The study enrolled patients who underwent lumbar pedicle screw fixation at West China Hospital of Sichuan University between March and September 2024. All patients underwent lumbar CT scans using a dual-layer detector system, and four image sets were reconstructed. CLumbar routine scans were performed using dual-layer detector CT, and four image sets were reconstructed:onventional images (CI non-O-MAR), conventional images with O-MAR (CI O-MAR), virtual monoenergetic images (VMI non-O-MAR), and VMI with O-MAR (VMI O-MAR). Objective metrics including artifact index (AI), signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) were quantified, alongside subjective image quality assessment. One-way ANOVA or Friedman test were used to compare the objective evaluation indicators of image quality between VMI non-O-MAR and VMI O-MAR at different energy levels. Paired t-test or Wilcoxon signed-rank test was used for CI non-O-MAR/VMI non-O-MAR versus CI O-MAR/VMI O-MAR comparisons. Results:A total of 30 patients were included, and all underwent internal fixation with titanium alloy pedicle screws. Objective analysis revealed that in both high-and low-density artifact regions, the AI values of the VMI O-MAR group decreased with the increase of energy levels, and were significantly lower than those of the corresponding VMI non-O-MAR group, with a statistically significant difference (all P<0.001). When the energy level≥140 keV, the AI value of the VMI O-MAR group was simultaneously lower than that of the CI non-O-MAR group and the CI O-MAR group, with statistically significant differences (all P<0.05). The SNR and CNR on the vertebral bodies in the VMI non-O-MAR group and the VMI O-MAR group showed a decreasing trend with increasing energy levels, and were always lower than those in the CI group at high energy levels (100-180 keV) (all P<0.05). At high energy levels (100-180 keV), the SNR of VMI O-MAR in the intervertebral disc and intraspinal tissues was higher than that of the VMI non-O-MAR group, with statistically significant differences (all P<0.05). Compared to other groups, subjective analysis indicated that the 140 keV VMI combined with O-MAR group received the highest image quality scores ( P<0.05). Conclusions:The combined application of VMI and O-MAR technology effectively reduces metal artifacts in post-lumbar fixation CT images. The 140 keV VMI with O-MAR reconstruction provides superior image quality and enhances diagnostic confidence.

Objective: To analyze the association of participation in non-sports extracurricular tutoring classes with the prevalence of screening myopia, axial length (AL) and axial length to corneal radius ratio (AL/CR) among primary school students, so as to provide evidences for formulating myopia prevention and control policies. Methods: In December 2024, combination of convenience and cluster sampling method was used to select 2 273 students from two primary schools in Hefei City, Anhui Province. Ophthalmic examinations and questionnaire surveys were conducted to obtain information on myopia, AL, AL/CR and participation in various types of extracurricular tutoring. A binary Logistic regression model was used to analyze the association between non-sports tutoring and screening myopia, and multiple linear regression models were used to examine the associations between non-sports tutoring and AL and AL/CR. Results: Among the surveyed students, the participation rate in non-sports extracurricular tutoring classes was 64.9% , and the overall prevalence of screening myopia was 39.1%. The average AL and AL/CR were (23.60± 1.01 ) mm and (3.00±0.12), respectively. Univariate analysis showed that students who attended non-sports, music, or academic tutoring classes for ≥2 h per week had higher risks of screening myopia and greater AL/CR values than non-participants (screening myopia: OR =1.38, 1.82, 1.55; AL/CR: β =0.01, 0.03, 0.03; all P <0.05). After adjusting for sex, grade, and participation in sports tutoring, multivariate analysis indicated that participation in non-sports and musical instrument tutoring classes for ≥2 h per week remained significantly associated with higher risks of screening myopia ( OR =1.26, 1.49, both P <0.05). Multiple linear regression showed that participation in musical instrument tutoring for ≥2 h per week was positively correlated with AL ( β=0.14, P < 0.05). Conclusions Participation in non-sports extracurricular tutoring is common among primary school students. Attending non-sports tutoring classes for ≥2 h per week increases the risk of screening myopia.

Objective To screen the anti-Streptococcus pneumoniae(Spn)active ingredients in vitro from different po-lar parts of Pilea peltata,and to examine the combined antibacterial effect of the active ingredient and amoxicillin(AMX).Methods A 96-well plate microdilution method was used to determine the minimum inhibitory concentration(MIC)of different polar parts;the most active polar part was separated by preparative high-performance liquid chromatography,and the active ingre-dients were identified using spectral technology.The fractional inhibitory concentration(FIC)of active ingredients and AMX was determined by the 96-well plate chessboard microdilution method.The crystal violet method was used to investigate the effect of ac-tive ingredients on Spn biofilm.The effect of active ingredients on the appearance and morphology of Spn was investigated under the electron microscope.Results The MICs of the petroleum ether part,chloroform part,ethyl acetate part,n-butanol part,and water part were 1.000,1.000,0.500,1.000,and 2.000 mg·mL-1 respectively,among which the ethyl acetate part had the stron-gest antibacterial activity.Three compounds were isolated from ethyl acetate,namely 5,7,3',4'-tetramethoxyflavone 1,8-O-(p-coumaroyl)-1(10)E,4(5)E-humuladien-8-ol 2 and 1-O-p-coumaroyl copaborneol 3.These compounds were all isolated for the first time from Pilea peltate,their MICs against Spn were 200.000,50.000,and 25.000 μg·mL-1 respectively,and the compound 3 had the strongest antibacterial activity;the FIC value of AMX and compound 3 was 0.50,which had a synergistic antibacterial effect on Spn.Both AMX and compound 3 had inhibitory effects on Spn biofilm,but the biofilm inhibition rate of compound 3(59.10±1.04％)was significantly lower than AMX(87.38±0.84)％(P＜0.01);Moreover,there was no significant difference in biofilm inhibition rate between the combination of the two and AMX(P＞0.05).The scanning electron microscope results showed that the bacterial cells in the compound 3 group had a smooth surface but varying degrees of depression.The surface of the bacteri-al cells in the AMX group and the AMX combined compound 3 group showed severe swelling and rupture.Conclusions Fla-vonoids and sesquiterpenoids are both the anti-Spn active components of Pilea peltate.Among them,sesquiterpenoids have more potent antibacterial activity,and their antibacterial action mechanism is related to inhibiting bacterial biofilms.Compound 3 and AMX have a synergistic antibacterial effect on Spn,but its mechanism of action is not by enhancing biofilm inhibition;although compound 3 cannot destroy the cell wall of Spn,it still has a negative impact on the appearance of the bacteria.