1.Arginine Metabolic Disorder in Heart Failure Rats: Analysis Based on Targeted Metabolomics and Bioinformatics
Zeyu LI ; Xiaoqing WANG ; Zhengyu FANG ; Yurou ZHAO ; He XIAO ; Penghaobang LIU ; Haiming ZHANG ; Chunyan LIU ; Yanhong HU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):229-237
ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure (HF), providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine (TCM). MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats, which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group, with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-S), and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging (AFADESI-MSI). Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus (GEO), a protein-protein interaction (PPI) network was constructed, and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) was performed. Finally, molecular docking was conducted to verify the binding between core metabolic components and key targets, and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group, the levels of arginine and citrulline in the serum of model rats were significantly decreased (P<0.01), while those of proline, ornithine, creatine, creatinine and glutamate were significantly increased (P<0.05, P<0.01). Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets, such as the angiotensin-converting enzyme (ACE), neuronal nitric oxide synthase (NOS1), 5-hydroxytryptamine receptor 2A (HTR2A), and epidermal growth factor receptor (EGFR), and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway, neuroactive ligand-receptor interactions, and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine, citrulline and HTR2A, as well as between creatine, creatinine and EGFR. Based on pathway-target prediction, potential TCM interventions, such as ginseng and magnolia, were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF, and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways, and offers valuable insights for targeted therapy of HF and the development of TCM.
2.Arginine Metabolic Disorder in Heart Failure Rats: Analysis Based on Targeted Metabolomics and Bioinformatics
Zeyu LI ; Xiaoqing WANG ; Zhengyu FANG ; Yurou ZHAO ; He XIAO ; Penghaobang LIU ; Haiming ZHANG ; Chunyan LIU ; Yanhong HU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):229-237
ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure (HF), providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine (TCM). MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats, which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group, with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-S), and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging (AFADESI-MSI). Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus (GEO), a protein-protein interaction (PPI) network was constructed, and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) was performed. Finally, molecular docking was conducted to verify the binding between core metabolic components and key targets, and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group, the levels of arginine and citrulline in the serum of model rats were significantly decreased (P<0.01), while those of proline, ornithine, creatine, creatinine and glutamate were significantly increased (P<0.05, P<0.01). Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets, such as the angiotensin-converting enzyme (ACE), neuronal nitric oxide synthase (NOS1), 5-hydroxytryptamine receptor 2A (HTR2A), and epidermal growth factor receptor (EGFR), and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway, neuroactive ligand-receptor interactions, and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine, citrulline and HTR2A, as well as between creatine, creatinine and EGFR. Based on pathway-target prediction, potential TCM interventions, such as ginseng and magnolia, were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF, and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways, and offers valuable insights for targeted therapy of HF and the development of TCM.
3.Risk factors for surgical site infection after colorectal cancer surgery: a two-center retrospective study
Zaihu MU ; Shuai ZHAO ; Wei CHEN ; Xiaoli YE ; Cong HAN ; Xiaojun JIN ; Aibin LIU ; Yanhong WENG ; Daorong WANG
Chinese Journal of Gastrointestinal Surgery 2025;28(10):1156-1160
Objective:To analyze the incidence of surgical site infection (SSI) in patients undergoing colorectal cancer (CRC) surgery and to identify risk factors associated with SSI in an attempt to provide a reference for clinical prevention strategies.Methods:A retrospective cohort study was conducted. Clinical data were retrospectively collected from a total of 2,248 patients who underwent surgery for pathologically confirmed CRC between 2017 and 2022 at two centers: Huangshan Shoukang Hospital ( n=649) and Northern Jiangsu People's Hospital ( n=1 599). Inclusion criteria consisted of the following: (1) age >18 years; (2) pathologically confirmed CRC treated with curative resection, including extended resections (e.g. pelvic exenteration); (3) no surgical incisions other than abdominal or perineal; and (4) no use of prosthetic implants. The incidence of SSI was analyzed, and multivariate logistic regression was used to identify independent its risk factors. Results:A total of 121 patients (5.4%) developed SSI. Among them, 68 cases (56.2%) were organ/space infections, 35 cases (28.9%) were deep incisional infections, and 18 cases (14.9%) were superficial incisional infections. The median postoperative hospital stay was significantly longer in patients with SSI compared to those without (21.0 days vs. 13.0 days, U=65,754, P<0.001). The median hospitalization cost was also significantly higher in the SSI group (56,550 yuan vs. 43,645 yuan, U=72,008, P<0.001). Multivariate logistic regression analysis identified body mass index (BMI) ≤ 20 kg/m 2 (OR=4.25, 95%CI: 3.38-5.34, P<0.001), diabetes mellitus (OR=3.44, 95%CI: 1.89-6.24, P<0.001), open surgery (OR=4.23, 95%CI: 2.37-7.56, P<0.001), and colostomy or ileostomy (OR=1.67, 95% CI: 1.04-2.69, P=0.034) as independent risk factors for SSI. Conclusion:To prevent SSI following CRC surgery, attention should be given to optimizing body weight and glycemic control, promoting minimally invasive surgical approaches when feasible, and cautiously considering the necessity of colostomy or ileostomy.
4.Mechanism of the Inhibitory Effect of Astragalus Polysaccharide on the Proliferation and Metastasis of Osteosarcoma Regulate by SP1/Wnt/β-catenin Signaling Axis
Ruizhong LI ; Yanhong LIN ; Chaohua LIANG ; Xinyu MA ; Shengsheng ZHAO ; Daochen LIANG
Journal of Modern Laboratory Medicine 2025;40(3):37-41,46
Objective To investigate the mechanism of astragalus polysaccharide(APS)in inhibiting the proliferation and metastasis of osteosarcoma cells.Methods Cell counting kit-8(CCK8)assay was used to detect the inhibitory effect of APS on the proliferation of osteosarcoma cell lines U2OS,HOS,MG63 and osteoblast cell line hFOB1.19,the cell line with the most significant inhibitory effect of APS on osteosarcoma cells was selected for subsequent experiments.Osteosarcoma cells were divided into control group(NC group),APS group,APS+sh-SP1 co treatment group with transfected SP1 knockdown plasmid(APS+sh-SP1),and APS+oe SP1 co treatment group with transfected SP1 overexpression plasmid(APS+oe SP1 group).Western blotting was used to detect the expression of SP1 protein in each group.CCK8 assay was used to detect the proliferation ability of each group.Transwell assay was used to detect the metastatic ability of cells in each group.TOP/FOP Flash assay was used to detect the activity of Wnt/β-catenin signaling pathway.The protein expressions of Wnt3a,β-catenin,CyclinD1,cMYC,matrix metalloproteinase 2(MMP2)and Snail were detected by western blotting.Result After 48h of APS treatment,the cell proliferation inhibition rates of osteosarcoma cells U2OS,HOS,MG63 and osteoblast cells hFOB1.19 were 62.93%±4.79%,20.66%±1.10%,39.31%±3.20%and 5.97%±0.72%,respectively.Compared with osteoblast hFOB1.19,APS significantly inhibited the proliferation of osteosarcoma cells,and the difference was statistically significant(F=208.400,P<0.001),and the inhibitory effect on osteosarcoma U2OS cells was the most significant(t=20.380,P<0.001).Compared with NC group,SP1 protein expression,cell proliferation ability,number of transmembrane cells,Wnt/β-catenin signaling pathway activity in cells,Wnt/β-catenin signaling pathway key proteins Wnt3a,β-catenin,downstream proliferation-related protein CyclinD1,cMYC,and downstream metastasis related proteins MMP2 and Snail in the APS group were decreased,and the differences were statistically significant(t=9.740~90.780,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+sh-SP1 group were further decreased,and the differences were statistically significant(t=3.032~12.940,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+oe-SP1 group were increased,and the differences were statistically significant(t=3.350~22.450,all P<0.05).Conclusion APS inhibits the proliferation and metastasis of osteosarcoma cells by targeting SP1/Wnt/β-catenin signaling axis.
5.The Guideline for the Diagnosis and Treatment of Pediatric Food Accumulation and Stagnation in Chinese Medicine
Xia ZHAO ; Mengting QIU ; Siyuan HU ; Yanhong QIN ; Zheng XUE ; Liqun WU
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(4):531-537
Following the evidence-based diagnosis and treatment guideline development method for common pediatric diseases,the The Guideline for the Diagnosis and Treatment of Pediatric Food Accumulation and Stagnation in Chinese Medicine was formed through literature retrieval,expert questionnaire survey,expert discussion meeting,and solicitation of opinions.The revised content includes the scope of the guideline,normative reference documents,terms and definitions,diagnosis,syndrome differentiation,treatment,pre-vention and nursing suggestions,etc.,aiming to update the clinical diagnosis and treatment plan of Chinese medicine for accumulation and provide important clinical decision-making references for clinical physicians to rationally apply the diagnosis and treatment plan of Chinese medicine in the prevention and treatment of accumulation and stagnation in Children.
6.NLUS-VQA: construction and evaluation of a visual question answering model for neonatal lung ultrasound diagnosis
Xuming TONG ; Jiangang CHEN ; Yiran WANG ; Xiqing ZHAO ; Yanhong YUAN ; Zishuo WANG ; Peng JIANG ; Qingyao XIONG ; Renxing LI ; Xueli WANG ; Jing LIU
Chinese Journal of Perinatal Medicine 2025;28(11):917-928
Objective:To develop and evaluate a medical visual question answering (VQA) model for neonatal lung ultrasound (LUS) images to enhance intelligent auxiliary diagnosis of neonatal pulmonary diseases.Methods:Using data from neonates admitted to Beijing Obstetrics and Gynecology Hospital, Capital Medical University (January 2023 to December 2024), an image-question-answer dataset comprising 251 LUS images was constructed [43 pneumonia (17.1%), 42 neonatal respiratory distress syndrome (16.7%), 83 transient tachypnea (33.1%), and 83 normal (33.1%) images] with a four-tier medical question-answer framework. Building upon the Qwen2.5-VL-7B base model and integrating LoRA fine-tuning with chain-of-thought prompting, we developed the NLUS-VQA model to enhance visual-language semantic alignment and enable stepwise clinical reasoning, achieving efficient small-sample adaptation. Model performance was comprehensively assessed through natural language generation metrics (BLEU-4, ROUGE-1/2/L), qualitative evaluation of characteristic recognition, and clinical consistency analysis.Results:(1) Quantitative evaluation demonstrated that NLUS-VQA achieved scores of 22.38 (BLEU-4), 48.26 (ROUGE-1), 22.40 (ROUGE-2), and 37.20 (ROUGE-L), representing significant improvements over baseline models. (2) Qualitatively, the model exhibited strong performance in identifying lung consolidation, coalescent B-lines, and snowflake signs, with its chain-of-thought strategy enhancing clinical interpretability and answer accuracy. (3) Clinically, NLUS-VQA achieved a Cohen's Kappa coefficient of 0.78 and diagnostic accuracy of 80.8% (21/26), indicating substantial agreement with clinical experts.Conclusion:The NLUS-VQA model demonstrates robust interpretability in recognizing key sonographic patterns (e.g. lung consolidation, confluent B-lines, and snowflake signs), providing a scalable framework for small-sample medical image analysis, though diagnostic performance on complex conditions remains limited by dataset scale and minority class representation.
7.Magnetic resonance imaging T2WI and DWI sequences in evaluating the therapeutic effect of neoadjuvant chemoradiotherapy for advanced rectal cancer
Ai CHENG ; Yanhong DONG ; Hui HUANG ; Wanqing LI ; Dacong ZHAO
Chinese Journal of Medical Physics 2025;42(6):801-805
Objective To explore the value of magnetic resonance imaging(MRI)T2-weighted imaging(T2WI)and diffusion-weighted imaging(DWI)sequences in evaluating the therapeutic effect of neoadjuvant chemoradiotherapy(nCRT)for advanced rectal cancer.Methods A retrospective analysis was conducted on 124 patients with advanced rectal cancer who received nCRT from November 2020 to November 2023 in Dazhou Integrated TCM&Western Medicine Hospital.All patients underwent radical resection of rectal cancer at 6-8 weeks after the end of nCRT.MRI scans were performed before nCRT and at 6-8 weeks after the end of nCRT.The differences in tumor long and short diameters,signal intensity in tumor(SIT),apparent diffusion coefficient(ADC)and other parameters were measured,and the MRI tumor regression grade was analyzed.Surgery was performed at 3 days after the second examination.The patients were divided into grade 0-1 group and grade 2-3 group according to the postoperative pathological tumor regression grade.The differences of MRI parameters before and after treatment were compared between groups.The consistency of T2WI sequence and DWI sequence in the diagnosis of tumor regression grade after nCRT was analyzed.ROC curve was used to analyze the value of MRI parameters in evaluating the therapeutic effect of nCRT.Results The differences of tumor long diameter,short diameter,SIT and ADC measured by twice MRI examination in grade 0-1 group were increased as compared with grade 2-3 group(P<0.05).MRI detected MRI tumor regression grade 1 in 28 cases(22.58%),grade 2 in 30 cases(24.19%),grade 3 in 31 cases(25.00%),grade 4 in 25 cases(20.16%)and grade 5 in 10 cases(8.06%).Consistency analysis showed that MRI had a sensitivity,specificity,accuracy rate and Kappa value of 92.45%,87.32%,89.51%and 0.788 in evaluating postoperative tumor regression grade.ROC analysis revealed that the AUC of MRI parameters such as differences in tumor long diameter,short diameter,SIT and ADC measured by twice MRI examinations in evaluating the nCRT efficacy ranged from 0.827 to 0.921,and the AUC of combined evaluation was 0.928(P<0.05).Conclusion The combined evaluation of MRI T2WI and DWI sequences can be served as an effective tool for evaluating the therapeutic effect of nCRT for advanced rectal cancer.
8.Comparison of the efficacy of ustekinumab and vedolizumab in achieving transmural healing in active Crohn′s disease
Long JU ; Yujie ZHAO ; Changqin LIU ; Yanhong SHI ; Xiaomin SUN ; Zhanju LIU ; Xiaolei WANG
Chinese Journal of Digestion 2025;45(2):107-116
Objective:To compare the efficacy of ustekinumab (UST) and vedolizumab (VDZ) in achieving transmural healing in active Crohn′s disease (CD).Methods:From March 1, 2020 to November 30, 2024, 112 patients with active CD treated with UST or VDZ at the Department of Gastroenterology, Tenth People′s Hospital of Tongji University were retrospectively enrolled. According to the medication regimen, the 112 patients were divided into UST group (61 cases) and VDZ group (51 cases). Collected the data at baseline, such as the disease phenotype, other medication history, and clinical indicators including C-creative protein (CRP), etc. Clinical disease activity and endoscopic disease activity were assessed by Harvey-Bradshaw index (HBI) and simplified endoscopic score for Crohn′s disease (SES-CD), respectively. Transmural healing was evaluated according to the intestinal wall thickness measured by intestinal imaging examination of the affected intestinal segment. Transmural healing was defined as bowel wall thickness <0.3 cm and 110 obvious signs of inflammation, clinical remission was defined as HBI≤4, and endoscopic remission was defined as a reduction in SES-CD ≥50% or a score of ≤2. The primary endpoint was transmural healing rate within one year after treatment. The secondary endpoints were endoscopic healing rate and clinical remission rate at 13 to 24th week and 30 to 52nd week after treatment. Chi-square test or Fisher′s exact test was used to compare the efficacy of the 2 medications.Results:There was no significant difference in transmural healing rate between UST group and VDZ group within 1 year after treatment (16.4% (10/61) vs. 23.5% (12/51), χ2=0.90, P=0.344). There were no significant differences in the healing rate between UST group and VDZ group in patients with specific baseline characteristics before treatment, including CD with stenosis, with perianal disease, no history of glucocorticoid use, secondary loss of response to anti-tumor necrosis factor (TNF)-α, SES-CD 7 to 15, decreased body mass index, and increased CRP (18.2%(6/33) vs. 19.4%(7/36), 17.9%(7/39) vs. 19.4%(6/31), 17.1%(6/35) vs. 24.2%(8/33), 20.0%(8/40) vs. 3/18, 14.3%(5/35) vs. 19.2%(5/26), 15.0%(3/20) vs. 3/10, 21.4%(6/28) vs. 5/16), all P>0.05). There was no significant difference in the clinical remission rate or endoscopic remission rate between the UST group and the VDZ group from 13 to 24th week (7/14 vs. 9/18, 3/14 vs. 7/18, RR=1.000 and 0.551, 95% confidence interval: 0.497to 2.011, 0.173to 1.755, χ2=<0.01, Fisher′s exact test, both P>0.05). There was no significant difference in clinical remission rate or endoscopic remission rate between UST group and VDZ group from week 30 to week 52 after treatment (68.5% (37/54) vs. 74.4% (32/43), 27.8% (15/54) vs. 32.6% (14/43), RR=0.921 and 0.853, 95% confidence interval: 0.716 to 1.184, 0.464 to 1.568, χ2=0.41 and 0.26, both P>0.05). In UST group, the proportion of patients with normal hemoglobin after transmural healing was higher than that of patients without transmural healing (9/10 vs. 45.1% (23/51), and the difference was statistically significant ( χ2=5.08, P=0.024). However, there were no significant differences in the proportion of patients with normal body mass index, CRP, platelet count, prealbumin, albumin, interleukin-6 or TNF-α levels after treatment between those who achieved transmural healing and those who did not in either UST group or VDZ group (all P>0.05). And in VDZ group there was no significant difference in the proportion of patients with normal hemoglobin between those who achieved transmural healing and who did not (all P>0.05). Conclusion:UST and VDZ exhibit similar efficacy in transmural healing within one year of treatment in patients with active CD.
9.Tissue-SELEX for screening nucleic acid aptamers targeting atherosclerotic tissue
Jianmin CHEN ; Zhimian SHI ; Yanhong LI ; Haiqing ZHAO ; Wenwang LANG ; Zhongcheng MO ; Jiangbo WANG
Chinese Journal of Arteriosclerosis 2025;33(11):937-943
Aim Systematic evolution of ligands by exponential enrichment(SELEX)techniquewas employed to screen and identify nucleic acid aptamers that specifically bind to mouse atherosclerotic pathological tissues,aiming to pro-vide a research foundation for the development of molecular targets and diagnostic reagents for early atherosclerosis.Methods A single-stranded DNA(ssDNA)library with a capacity of 1015~1016 was constructed,which was then subjec-ted to binding-elution(negative selection)with normal mouse vascular tissue slices.The eluted library was subsequently bound to atherosclerotic tissue slices for binding-elution(positive selection).PCR was used to amplify the positive and negative screening products,and agarose gel electrophoresis was used to verify the amplified products.The ssDNA library after multiple rounds of selection was sequenced using T-A cloning and sequencing to obtain the primary structure of the nu-cleic acid aptamers,and the secondary structure was predicted using the Mfold online software.The selected nucleic acid aptamers were labeled with a FAM fluorescent group at the 5'-end and were bound to both positive and negative selection tissue slices,with fluorescence intensity observed under a fluorescence microscope.Image Pro Plus 6.0 was used to cal-culate the relative average fluorescence intensity to evaluate the binding specificity of nucleic acid aptamers.Results After 8 rounds of selection,agarose gel electrophoresis imaging showed PCR amplification products in the positive selection lanes,while no PCR amplification products were observed in the negative selection lanes,indicating the successful acquisi-tion of a nucleic acid aptamer library that specifically binds to atherosclerotic tissues.Five nucleic acid aptamers were i-dentified by T-A cloning and sequencing,and their predicted secondary structures all had stem-loop structures.Immuno-fluorescence staining verified that five nucleic acid aptamers had different degrees of binding with As blood vessels,and the quantitative results of the relative average fluorescence intensity showed that nucleic acid aptamer No.11 had the highest relative average fluorescence intensity value,which can be used as a candidate nucleic acid aptamer for subsequent re-search.Conclusion Specific nucleic acid aptamers that bind to atherosclerotic vesselswere successfully obtained,providing a research foundation for further screening of early molecular targets of Asand developing in vivo early diagnostic reagents.
10.Mechanism of the Inhibitory Effect of Astragalus Polysaccharide on the Proliferation and Metastasis of Osteosarcoma Regulate by SP1/Wnt/β-catenin Signaling Axis
Ruizhong LI ; Yanhong LIN ; Chaohua LIANG ; Xinyu MA ; Shengsheng ZHAO ; Daochen LIANG
Journal of Modern Laboratory Medicine 2025;40(3):37-41,46
Objective To investigate the mechanism of astragalus polysaccharide(APS)in inhibiting the proliferation and metastasis of osteosarcoma cells.Methods Cell counting kit-8(CCK8)assay was used to detect the inhibitory effect of APS on the proliferation of osteosarcoma cell lines U2OS,HOS,MG63 and osteoblast cell line hFOB1.19,the cell line with the most significant inhibitory effect of APS on osteosarcoma cells was selected for subsequent experiments.Osteosarcoma cells were divided into control group(NC group),APS group,APS+sh-SP1 co treatment group with transfected SP1 knockdown plasmid(APS+sh-SP1),and APS+oe SP1 co treatment group with transfected SP1 overexpression plasmid(APS+oe SP1 group).Western blotting was used to detect the expression of SP1 protein in each group.CCK8 assay was used to detect the proliferation ability of each group.Transwell assay was used to detect the metastatic ability of cells in each group.TOP/FOP Flash assay was used to detect the activity of Wnt/β-catenin signaling pathway.The protein expressions of Wnt3a,β-catenin,CyclinD1,cMYC,matrix metalloproteinase 2(MMP2)and Snail were detected by western blotting.Result After 48h of APS treatment,the cell proliferation inhibition rates of osteosarcoma cells U2OS,HOS,MG63 and osteoblast cells hFOB1.19 were 62.93%±4.79%,20.66%±1.10%,39.31%±3.20%and 5.97%±0.72%,respectively.Compared with osteoblast hFOB1.19,APS significantly inhibited the proliferation of osteosarcoma cells,and the difference was statistically significant(F=208.400,P<0.001),and the inhibitory effect on osteosarcoma U2OS cells was the most significant(t=20.380,P<0.001).Compared with NC group,SP1 protein expression,cell proliferation ability,number of transmembrane cells,Wnt/β-catenin signaling pathway activity in cells,Wnt/β-catenin signaling pathway key proteins Wnt3a,β-catenin,downstream proliferation-related protein CyclinD1,cMYC,and downstream metastasis related proteins MMP2 and Snail in the APS group were decreased,and the differences were statistically significant(t=9.740~90.780,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+sh-SP1 group were further decreased,and the differences were statistically significant(t=3.032~12.940,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+oe-SP1 group were increased,and the differences were statistically significant(t=3.350~22.450,all P<0.05).Conclusion APS inhibits the proliferation and metastasis of osteosarcoma cells by targeting SP1/Wnt/β-catenin signaling axis.

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