To evaluate the application value of the Peyton four-step teaching method in the standardized training of intestinal ultrasound and compare it with traditional teaching methods, so as to provide an optimized approach for clinical ultrasound training.

ObjectiveThis study aimed to establish a monocrotaline （MCT）-induced pulmonary arterial hypertension （PAH） rat model to systematically evaluate the protective effect of Xiao Qinglongtang （XQLT） on right cardiac function in model rats and further elucidate the underlying regulatory mechanism. MethodsSixty male SD rats were randomly assigned to the normal group， model group， XQLT low-， medium-， and high-dose groups （XQLT-L/M/H）， and the beraprost sodium tablet group （BST）. Except for the normal group， rats in all other groups were given a single subcutaneous injection of MCT （60 mg·kg^-1） to induce PAH. Three weeks after injection， rats in the XQLT-L/M/H groups were administered XQLT intragastrically at 3.07， 6.14， 12.28 g·kg^-1·d^-1， respectively. Rats in the BST group received beraprost sodium at 12.6 μg·kg^-1·d^-1， and rats in the model group received an equal volume of saline. All treatments lasted for 3 weeks. Right ventricular systolic pressure （RVSP） was measured by right ventricular catheterization. Cardiac function was assessed by echocardiography. The right ventricle was weighed to calculate the right ventricular hypertrophy index （RVHI）. Hematoxylin-eosin （HE） staining， Masson staining， and transmission electron microscopy were used to observe myocardial morphology. Serum metabolomic changes were analyzed using ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）. Data-independent acquisition （DIA） proteomics was used to detect differentially expressed （DE） proteins in the right ventricle， and Western blot was used to measure the expression of uncoupling protein 3 （UCP3）， phosphatidylinositol 3-kinase catalytic subunit p110α （PIK3CA）， L1 cell adhesion molecule （L1CAM）， and quinone oxidoreductase （CRYZ）. UPLC-MS/MS was used to analyze the chemical components of XQLT. ResultsCompared with the normal group， the model group showed significantly increased RVSP and RVHI （P<0.05）， along with pathological changes in myocardial morphology. Compared with the model group， all XQLT-treated groups exhibited reductions in RVSP and RVHI as well as significant improvements in cardiac function and myocardial morphology. Among the XQLT groups， XQLT-M showed the most pronounced effects （P<0.05）， comparable to the BST group. Serum metabolomics revealed 105 differential metabolites in the XQLT groups versus the model group ［variable importance in projection （VIP） >1， P<0.05］， including 58 upregulated and 47 downregulated metabolites. KEGG enrichment analysis indicated that XQLT intervention downregulated phenylalanine metabolism （P<0.01） and upregulated unsaturated fatty acid biosynthesis （P<0.05）. Proteomics analysis showed that 982 DE proteins were identified in the MCT groups versus the normal group， including 455 upregulated and 527 downregulated proteins （|fold change （FC）| >1.3， P<0.05）. Compared with the model group， 237 DE proteins were identified in the XQLT groups， including 124 upregulated and 113 downregulated proteins （|FC| >1.3， P<0.05）， with 57 overlapping DE proteins. KEGG enrichment suggested that XQLT mainly modulated pathways related to mineral absorption， ribosomal biogenesis， peroxisomes， glycolysis/gluconeogenesis， spliceosomes， and thyroid hormone signaling. Western blot analysis showed that， compared with the model group， XQLT increased the expression of UCP3， PIK3CA， and L1CAM， while decreasing the expression of CRYZ （P<0.05）. ConclusionXQLT exerts a protective effect on right heart function in MCT-induced PAH rats， and its mechanism is associated with maintaining myocardial homeostasis and alleviating right ventricular remodeling.

Background/Aims: Lenvatinib resistance remains a critical barrier in advanced hepatocellular carcinoma (HCC) therapy. However, the underlying mechanisms and strategies for reversing resistance remain incompletely understood. Methods: Integrated transcriptomics of lenvatinib-resistant patient tumors and an acquired-resistance murine model identified a novel macrophage subpopulation. Functional validation employed CRISPR-SAM screening, conditioned medium (CM) assays, subcutaneous/orthotopic xenografts, patient-derived organoids (PDOs), and patient-derived xenografts (PDXs). Mechanistic studies included ChIP-qPCR, co-immunoprecipitation, and pharmacologic targeting. Clinical relevance was assessed in a retrospective cohort. Results: Resistant HCC exhibited significant enrichment of a COLEC12high TAM subset , which correlated with poor survival and treatment response. These TAMs secreted neuregulin-1 (NRG1) , activating HER2/HER3-AKT signaling in tumor cells to drive cancer stemness and lenvatinib resistance. Mechanistically, in TAMs COLEC12 sequestered STAT1 in the cytoplasm, preventing its phosphorylation, and thereby derepressing STAT3-mediated NRG1 transcription. Depletion of NRG1 reversed the stemness phenotypes and resensitized tumors to lenvatinib both in vitro and in vivo. Clinically, high NRG1 expression predicted an inferior lenvatinib response and shorter survival. Crucially, the bispecific anti-HER2/HER3 antibody zenocutuzumab restored lenvatinib efficacy in PDOs, PDXs, and murine models. Conclusions Our work establishes the COLEC12high TAM/NRG1 axis as a master regulator of therapeutic resistance and identifies NRG1 as a predictive biomarker, providing a clinically actionable strategy to overcome lenvatinib resistance in HCC.

ObjectiveTo investigate the physical fitness status and associated factors among preschool children aged 3 to 6 years in Tongzhou District of Beijing, and to provide evidence for promoting physical development in this population. MethodsBased on the 2020 National Physical Fitness Surveillance data for Tongzhou District of Beijing, a total of 416 children were included for standardized physical tests, including morphological, physiological, and fitness indicators, and questionnaire surveys. Independent sample t-tests and ANOVA were used to compare differences in physical indicators across groups, while multivariate linear regression models were applied to analyze influencing factors. ResultsThe comprehensive fitness score of children aged 3 to 6 years old in Tongzhou District of Beijing was (79.98±6.97) points, with an overall physical fitness pass rate of 99.52%. The proportions of excellent, good, and pass grades were 37.50%, 39.90%, and 22.12%, respectively. The average heights for the 3-years-old, 4-years-old, 5-years-old, and 6-years-old groups in Tongzhou District were (102.13±4.24) cm, (107.92±4.66) cm, (115.43±4.70) cm, and (119.16±4.33) cm, respectively. The corresponding average body weights were (16.63±2.46) kg, (18.66±2.54) kg, (21.60±4.02) kg, and (23.46±4.64) kg, respectively. Among children aged 3 to 6 years, the average BMI was (15.95±1.79) kg‧m^-2, body fat percentage was (21.92±4.77)%, resting pulse rate was (95.24±9.70) beats per minute, grip strength was (5.74±2.07) kg, standing long jump distance was (78.12±19.95) cm, sit-and-reach flexibility was (12.25±4.65) cm, continuous two-foot jump time was (6.29±2.55) seconds, 15-meter obstacle shuttle run time was (7.94±1.30) seconds, and balance beam walking time was (5.11±2.25) seconds. Multiple linear regression analyses showed that higher composite fitness scores were significantly positively associated with female gender (b=1.72, 95%CI: 0.42-3.01), aged 4 years old and above (b=2.97, 95%CI: 1.35-4.59), mixed feeding (b=4.16, 95%CI: 1.60-6.72) or breastfeeding (b=3.23, 95%CI: 0.36-6.10) during infancy, taller paternal (b=0.16, 95%CI: 0.02-0.30) and maternal (b=0.25, 95%CI: 0.11-0.39) height (all P<0.05). ConclusionThe physical fitness of children aged 3 to 6 years in Tongzhou District of Beijing is generally favorable, with gender, age, feeding patterns, and genetic factors identified as key correlating factors. Comprehensive strategies are recommended to improve early childhood physical health.

To investigate the effectiveness of a teaching model combining real-case bedside instruction with mixed-level group teaching in intestinal ultrasound training, and to provide reference forestablishing an efficient and practical training system for intestinal ultrasound.

ObjectiveTo establish a semi-automated workflow for hyoid motion tracking based on submental ultrasound videos, evaluate the reliability of manual measurements and the validity of semi-automated tracking, and compare the analytical efficiency. MethodsA total of 46 healthy adults were recruited at Xiamen Fifth Hospital from October, 2024 to August, 2025, and performed dry swallows and swallows of 5 mL and 10 mL moderately-thick liquid boluses. A total of 414 sagittal-plane submental ultrasound videos were acquired. After video quality screening and tracking quality-control procedures, 286 videos were included for paired analysis. A semi-automated tracking pipeline was constructed combining the channel-and-spatial-reliability tracker and You Only Look Once version 8 small (YOLO v8s). Intra-class correlation coefficients [ICC(A,1)] based on the absolute-agreement single-measurement model were calculated for test-retest and inter-rater manual measurements. Using the manual repeated-measurement results as the reference standard, Pearson correlation coefficient (r), ICC(A,1) and concordance correlation coefficient (CCC) were computed for the semi-automated measurements. A total of 5 000 subject-clustered bootstrap resamples were performed to calculate 95% confidence intervals. Cumulative analysis time of manual and semi-automated workflows was compared. ResultsFor the independent test set of YOLOv8s, precision and recall reached 0.940 and 0.956. The mean average precision was 0.974 at an intersection-over-union threshold of 0.5, and 0.609 averaged across thresholds ranging from 0.5 to 0.95. For manual measurements of maximum horizontal and vertical hyoid displacement, the test-retest ICC(A,1) values were 0.925 and 0.912, respectively; the inter-rater ICC(A,1) values were 0.815 and 0.775. For semi-automated versus manual reference measurements, r = 0.865，ICC(A,1) = 0.843，CCC = 0.843 in maximum horizontal displacement; and r = 0.754，ICC(A,1) = 0.746，CCC = 0.745 in maximum vertical displacement. Mean biases were -0.772 mm and -0.229 mm, with 95% limits of agreement of -4.567 to 3.023 mm and -2.918 to 2.460 mm for horizontal and vertical directions. Cumulative manual analysis time was 526 minutes for rater A and 713 minutes for rater B. The semi-automated workflow consumed 155 minutes, in which 40 minutes for computational processing and 115 minutes for manual review. ConclusionManual measurement of maximum hyoid displacement is reliable using qualified submental ultrasound videos. Semi-automated tracking achieves acceptable agreement with manual reference and greatly reduces analysis workload.

A novel analytical method was developed in this study by combining online solid phase extraction with ultra performance liquid chromatography-tandem mass spectrometry(Online SPE-UPLC-MS/MS)for simultaneous determination of 50 kinds of steroid hormones in surface water.Specifically,after high-speed centrifugation of 4 mL water samples,the supernatant was directly injected into an Oasis HLB online SPE column for enrichment and purification.Subsequently,the target compounds were transferred to the analytical column via valve switching for separation and analysis.The chromatographic separation was performed on a Thermo Acclaim RSLC C18 column(100 mm×2.1 mm,2.2 μm),using a mobile phase composed of 5 mmol/L ammonium fluoride aqueous solution and acetonitrile.Mass spectrometric detection was conducted in positive ion mode,utilizing multiple reaction monitoring(MRM)with quantification achieved by the internal standard method.The method validation demonstrated that the limits of detection(LOD)for the 50 kinds of steroid hormones ranged from 0.02 to 0.50 ng/L,while the limits of quantification(LOQ)were between 0.08 and 1.67 ng/L.The average recoveries in surface water samples at spiked concentrations of 5,20 and 200 ng/L were between 74.1％and 119％,with relative standard deviations(RSDs)of 0.2％to 9.9％.This method was applied to analyze 11 surface water samples collected from sites surrounding a pharmaceutical and chemical industrial park.A total of 44 kinds of steroid hormones were detected,with concentrations ranging from 0.11 to 88.6 ng/L,revealing the presence of hormone contamination in the environmental waters surrounding industrial areas.Compared with the traditional offline SPE methods,the proposed online SPE technique significantly reduced sample volume requirements and pretreatment time,while minimizing the loss of target compounds during the pretreatment process.Moreover,compared to reported online SPE techniques,this method achieved high-throughput analysis of multiple classes of steroid hormones,with lower detection limits and higher recoveries.Overall,this method provided rapid sample preparation,high sensitivity,and excellent stability,making it suitable for the direct analysis of trace steroid hormones in surface water.

Objective To investigate the effect of human umbilical vein endothelial cells(HUVECs)on the proliferation and stem-ness of human dental pulp stem cells(hDPSCs)silencing with integrin α6(ITGA6).Methods ITGA6 silencing lentivirus was used to interfere the ITGA6 expression of hDPSCs,and its silencing efficiency was detected.Then HUVECs were added to the chambers to co-culture,and the experiments were divided into four groups(sh-NC,sh-ITGA6,sh-NC+HUVECs and sh-ITGA6+HUVECs).hDP-SCs in the sh-NC and sh-ITGA6 groups were transfected with sh-NC and sh-ITGA6 respectively.hDPSCs transfected with sh-NC and sh-ITGA6 were co-cultured with HUVECs in the sh-NC+HUVECs group and sh-ITGA6+HUVECs group respectively.The proliferation capacity of hDPSCs of each group was examined by CCK-8 and EdU on day 7.Immunofluorescence detected the expression of Stro-1,and Real-time PCR was used to detect the expression of Oct4 and Nanog.Results ①Fluorescence microscopy showed that the trans-fection efficiency was about 80％.Real-time PCR and Western blot results showed that sh-ITGA6 lentivirus effectively interfered with ITGA6 expression in hDPSCs.②CCK-8 results showed that on day 5 of co-culture,the proliferation ability of the sh-ITGA6+HUVECs group was superior to that of the sh-ITGA6 group(P＜0.05);on day 7,the proliferation ability of the sh-NC+HUVECs and sh-ITGA6+HUVECs group was superior to that of the sh-NC and sh-ITGA6 group(P＜0.05).EdU results showed that the DNA synthesis ability of hDPSCs in the co-culture group was significantly stronger than that in the single-culture group(P＜0.05).③Immunofluorescence stai-ning revealed that the expression of Stro-1 in the co-culture group was significantly stronger than that in the single-culture group.④Re-al-time PCR results showed that the expression of Oct4 in the co-culture group was higher than that in the single-culture group(P＜0.05);the expression of Nanog in hDPSCs with sh-ITGA6 was elevated by the addition of HUVECs co-culture(P＜0.05).Conclusion HUVECs significantly enhance the proliferation and stemness of hDPSCs silencing integrin α6.

The chemical constituents were systematically separated from the roots of Berberis polyantha by various chromatographic methods, including silica gel column chromatography, HP20 column chromatography, polyamide column chromatography, reversed-phase C_(18) column chromatography, and preparative high-performance liquid chromatography. The structures of the compounds were identified by physicochemical properties and spectroscopic techniques(1D NMR, 2D NMR, UV, MS, and CD). Four phenylpropanoids were isolated from the methanol extract of the roots of B. polyantha, and they were identified as(2R)-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone-O-β-D-glucopyranoside(1), methyl 4-hydroxy-3,5-dimethoxybenzoate(2),(+)-syringaresinol(3), and syringaresinol-4-O-β-D-glucopyranoside(4). Compound 1 was a new compound, and other compounds were isolated from this plant for the first time. The anti-inflammatory activity of these compounds was evaluated based on the release of nitric oxide(NO) in the culture of lipopolysaccharide(LPS)-induced RAW264.7 macrophages. At a concentration of 10 μmol·L~(-1), all the four compounds inhibited the LPS-induced release of NO in RAW264.7 cells, demonstrating potential anti-inflammatory properties.
Plant Roots/chemistry* ; Animals ; Mice ; Berberis/chemistry* ; RAW 264.7 Cells ; Macrophages/immunology* ; Drugs, Chinese Herbal/isolation & purification* ; Nitric Oxide/metabolism* ; Molecular Structure ; Anti-Inflammatory Agents/isolation & purification*

Objective To investigate the expression level of centromere protein(CENP)Ⅰin lung adenocarcinoma cells,to study the effects of CENPⅠon the proliferation,invasion,migration,apoptosis,and epithelial-mesenchymal transition(EMT)of lung adenocarci-noma cells,and to explore the possible mechanisms related to its occurrence.Methods The expression of CENPⅠmRNA and protein in four types of lung adenocarcinoma cells and normal alveolar epithelial cells were detected by quantitative real-time polymerase chain reaction(RT-qPCR)and Western blotting.The expression of CENPⅠin H1650 cells was knocked down by the siRNA technique,The transfection efficiency was detected by RT-qPCR and Western blotting.The effects of knock-down CENPⅠon proliferation,cell cycle,apoptosis,invasion and migration of H1650 cells were detected by the cell counting kit-8 assay,the transwell assay,and flow cytometry.Western blotting was used to detect the expression of E-cadherin,N-cadherin,vimentin,Ki-67,cyclin D1,Bcl-2,PI3K,AKT,mTOR,p-PI3K,p-AKT,and p-mTOR.Results After the knock-down of CENPⅠ,the proliferative ability of the H1650 cells significantly decreased,the number of apoptotic cells significantly decreased,and the cell invasion and migration abilities significantly decreased(P<0.01).E-cadherin expression was upregulated and N-cadherin,vimentin,Ki-67,cyclin D1,Bcl-2,p-PI3K,p-AKT,and p-mTOR expres-sion were down-regulated in the CENTI group compared with the control group(P<0.05).The expression of p-PI3K,p-AKT,and p-mTOR in the si-CENPⅠ+IGF-1 group was upregulated compared to that in the si-CENPⅠgroup(P<0.05).Conclusion High expression of CENPⅠin lung adenocarcinoma cells promotes the proliferation,invasion,and migration of lung adenocarcinoma H1650 cells and EMT inhibits apoptosis,which may be related to the activation of the PI3K/AKT/mTOR signaling pathway.