1.Study on the effect of high-fidelity intelligent simulator combined with scenario simulation in emergency response training of radiology department
Zhengting ZHU ; Yuping ZHENG ; Manli CHENG ; Yang LIU ; Xueqiu YAN ; Li REN ; Haibo QU ; Huayan XU ; Yun WANG ; Gang NING
Chinese Journal of Medical Education Research 2025;24(9):1158-1163
Objective:To explore the application effect of high-fidelity intelligent simulator combined with scenario simulation for emergency response training in the Department of Radiology, and to improve the emergency preparedness of medical, nursing, and technical staff in managing contrast agent adverse reactions.Methods:From January to July 2024, 132 medical, nursing, and technical staff from the Department of Radiology of a tertiary hospital in Chengdu City, China were selected as the training subjects. The high-fidelity intelligent simulator combined with scenario simulation teaching mode was used to conduct emergency response training for the participants. The differences in theoretical knowledge and post competence regarding contrast agent adverse reactions among the staff were compared before and after the training. A self-made questionnaire was used to investigate their needs and satisfaction of the emergency response training. SPSS 26.0 was used for data analysis. The differences in theoretical knowledge and post competence scores before and after training were compared using the paired samples t test. Results:After the training, the average score of theoretical knowledge examination increased from (84.32±10.19) points to (90.34±7.87) points, and the difference was statistically significant ( P<0.001). After the training, the scores of knowledge reserve, operational skills, situational decision-making ability, professional literacy, comprehensive literacy, and overall post competency were all significantly higher than those before the training ( P<0.05). The satisfaction score of emergency response training was (4.17±0.25) points. Conclusions:High-fidelity intelligent simulator combined with scenario simulation training improved the emergency preparedness and teamwork of radiology staff in clinical emergencies. The training received high recognition and satisfaction from the participants, which is of great significance for clinical emergency response and patient safety.
2.Analyzing correlations of volume percentage and metabolism of brain region in patients with Alzheimer disease based on FDG PET/MR
Yinyan ZHU ; Yan ZHANG ; Gan HUANG ; Mei XIN ; Peizhe YUAN ; Yue WANG ; Liangrong WAN ; Cheng WANG ; Gang HUANG ; Jianjun LIU ; Chenpeng ZHANG
Chinese Journal of Medical Imaging Technology 2025;41(2):203-206
Objective To investigate correlations of the ratio of specific brain region volume to total brain volume(ratio%),standard uptake value(SUV)of specific brain region based on 18F-FDG PET/MR examination in Alzheimer disease(AD)patients,as well as the relationship between changes of these two.Methods Fifty AD patients were retrospectively collected.Based on FDG PET/MR,the ratio%and the mean SUV(SUVmean)of 40 specific brain regions were obtained,and the correlation between ratio%and SUVmean for each region were evaluated.According to a database of normal brains,Z-scores for ratio%and SUVmean were calculated to represent structural and functional changes in AD patients,and the correlation between these 2 Z-scores was assessed.Results Correlations were found between ratio%and SUVmean,also between Z-scores of these two parameters in 29 brain regions(r=0.288 to 0.778,all P<0.05).Among them 7 brain regions,i.e.bilateral middle temporal gyrus,right fusiform gyrus,right hippocampus,right precuneus,left lingual gyrus and right parahippocampal gyrus exhibited correlation coefficients greater than 0.6.Conclusion There were some relationships between brain structural and metabolic functions and their changes in AD patients showed on FDG PET/MR.
3.Chemical constituents from Asteris Radix et Rhizoma and their anti-inflammatory activities
Pei LI ; Fu-xia ZHAO ; Si-qi YANG ; Jin-yan TAN ; Ying-li WANG ; Yan-gang CHENG
Chinese Traditional Patent Medicine 2025;47(2):473-479
AIM To study the chemical constituents from Asteris Radix et Rhizoma and their anti-inflammatory activities.METHODS The extract from Asteris Radix et Rhizoma was isolated and purified by column chromatography and high performance liquid chromatography,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activities were evaluated by RAW264.7 model.RESULTS Thirteen compounds were isolated and identified as(Z)-9,10,11-trihydroxy-12-octadecenoic acid(1),tianshic acid(2),6,6-dimethyl-2-methlenebicyclo[3.1.1]hept-3-O-(6-O-apiofuranosyl)-β-D-glucopyranoside(3),ent-16β,17-dihydroxy-kauran-19-oic acid(4),ent-17-hydroxy-19-kauranoic acid(5),7β,17-dihydroxy-16α-ent-kauran-19-oic acid 19-O-β-D-glucopyranoside ester(6),paniculoside Ⅳ(7),thomimarine A(8),cyclo-(S-Pro-R-Leu)(9),4,5-di-O-caffeoylquinic acid 1-methyl ether(10),methyl 3,5-di-O-caffeoyl quinate(11),5-acetyl-3β-hydroxy-2β-(1-hydroxyisopropyl)-2,3-dihydrobenzofurane(12),4-ally-2,6-dimethoxyphenyl glucoside(13).Compounds 1 and 3-12 had inhibition on the release of NO in RAW264.7 cells,and 4-6,8,10-12 were better than the positive control.CONCLUSION Compounds 1,6,8-9 are isolated from Compositae family for the first time,and 2-5,7,10 and 11-13 are first isolated from this plant.Compounds 1,3-12 have anti-inflammatory activities.
4.Application of ultrasound-guided injection of carbon nanoparticle in cervical lymph node dissection for thyroid cancer reoperation
Yue ZHOU ; Gang WANG ; Fang YU ; Hao XU ; Zhenyu CHENG ; Ziyi FAN ; Xianjiao CAO ; Zhonghui LI ; Qingqing HE
Chinese Journal of Endocrine Surgery 2025;19(1):30-34
Objective:To investigate the efficacy of preoperative ultrasound-guided injection of carbon nanoparticle suspension in cervical lymph node dissection for thyroid cancer reoperation.Methods:Ninety-four patients undergoing reoperation for thyroid cancer admitted by the same physician team of the Department of Thyroid and Breast Surgery of the Ninety-sixty Hospital of the People’s Liberation Army (PLA) from Jan. 2019 to Sep. 2024 were retrospectively analyzed. According to the different scope of the initial surgery, they were divided into the re-specification clearance group, the regional lymph node clearance group, and the metastatic lymph node dissection group, and the groups were subdivided into the carbon nanoparticle group and the control group according to whether they were injected with carbon nanoparticle before the surgery or not. The t-test, χ2-test, and non-parametric test were used to compare the age, gender, surgical method, duration of surgery, total number of lymph nodes detected, and positive lymph node detection rate between the nano-charcoal group and the control group in the three groups. Results:There was no statistically significant difference between the three groups in terms of age, gender, surgical methods, or the total number of lymph nodes detected (all P>0.05) , and the difference between the surgical time of the carbon nanoparticle group in the re-regulation clearance group and the control group was not statistically significant ( P>0.05) ; the surgical time of the carbon nanoparticle group was shorter than that of the control group in both the regional lymph node clearance and the metastatic lymph node dissection groups, and the difference was statistically significant (all P<0.05) ; the positive detection rate of lymph nodes in the carbon nanoparticle group was higher than that in the control group among the three groups, and the difference was statistically significant (all P<0.05) . Conclusion:In the operation of cervical lymph node dissection for papillary thyroid cancer, preoperative ultrasound-guided injection of carbon nanoparticle can accurately localize the lymph nodes, increase the positive detection rate of lymph nodes, reduce the difficulty of surgical operation, and shorten the operation time.
5.Effect of intensive transcranial magnetic stimulation on cognitive function and systemic immune-inflammation index in patients with treatment-resistant depression
Qi WANG ; Wenwen CHENG ; Xiaomei DONG ; Zhongli GENG ; Gang CUI ; Lin FAN ; Tianchao XU
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(1):30-35
Objective:To explore the efficacy of intensive repetitive transcranial magnetic stimulation (irTMS) in treatment-resistant depression (TRD) and its impact on cognitive function and systemic immune-inflammation index (SII).Methods:Forty-eight TRD patients were divided into observation group and control group using random number table method, with 24 patients in each group. The observation group was treated with irTMS, and the stimulation site was the left dorsolateral prefrontal lobe. The stimulation intensity was 110% of the motor threshold, and the stimulation frequency was 15 Hz. The stimulation interval was 26 s, and 3 000 pulses were stimulated each time. Stimulating 5 times per day, with an interval of 50 min, and continuous treatment for 5 days. The total stimulation amount for 5 days was 75 000 pulses. The control group was treated with pseudo stimulation. Before treatment (T0), 5 days after treatment (T1), and 1 month after treatment (T2), 17-item Hamilton depression scale (HAMD-17) was used to assess depressive mood. Evaluating cognitive function using the Wisconsin card sorting test.A fully automated blood cell analyzer was used to detect platelet count (PLT), neutrophil count (NC), and lymphocyte count (LC), calculate the systemic immune inflammation index (SII), SII=PLT × NC/LC. Statistical analysis was conducted using SPSS 20.0 software. The comparison between two sets of repeated measurement data was performed using repeated measurement analysis of variance.Simple effect analysis was performed if the interaction effect was significant.Pearson analysis was used for correlation testing.Results:The interaction effect between the time and group of HAMD-17 scores was significant ( F=121.784, P<0.05). The results of simple effects analysis showed that the HAMD-17 scores of the observation group at T1 and T2 ((12.07±4.08) and (14.78±4.99), respectively) were lower than those of the control group ((23.78±5.87) and (24.67±7.00), P<0.05). The treatment response rate and remission rate of the observation group at T1 were higher than those of the control group ( χ2=4.090, 7.378, both P<0.05).The treatment response rate and remission rate of the observation group at T2 were higher than those of the control group ( χ2=4.463, 4.547, both P<0.05). The time and group interaction effects of the percentage of correct response and conceptualization level in the Wisconsin card sorting test were significant ( F=30.087, 20.004, P<0.05). The results of simple effects analysis showed that the percentage of correct response and conceptualization level in the observation group at T1 and T2 were higher than those in the control group (all P<0.05). The time and group interaction effect of SII was significant ( F=8.173, P<0.05). The results of simple effects analysis showed that there was no statistically significant difference in SII between the two groups at T0 ( P>0.05). The SII at T1 and T2 in the observation group was lower than that in the control group ( P<0.05). In the observation group, the changes in SII from T2 to T0 was positively correlated with the change in HAMD-17 scores ( r=0.527, P<0.05), and negatively correlated with the percentage of correct responses to the Wisconsin card sorting test ( r=-0.412, P<0.05) and the percentage of conceptualization level ( r=-0.411, P<0.05). Conclusion:irTMS is effective in treating TRD and can improve patients' cognitive function and immune inflammation damage.
6.The role of SIRT7 in regulating epithelial-mesenchymal transition(EMT)to promote glioma cell proliferation and migration
Jiang-Ting WANG ; Kai SUN ; Mou GAO ; Gang CHENG ; Wen-Ying LYU ; Chun-Hui ZHOU ; Lei LIANG ; Jian-Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2025;50(1):57-68
Objective To investigate the expression of the histone deacetylase SIRT7 in glioma cells and its impact on epithelial-mesenchymal transformation(EMT),as well as its effects on proliferative,migratory and invasive capabilities of glioma cells.Methods Bioinformatics analysis was conducted on data from glioma patients in the Cancer Genome Atlas(TCGA)and the Chinese glioma Genome Atlas(CGGA)databases to explore the expression of SIRT7 gene in gliomas and its correlation with tumor grading,molecular characteristics and patient clinical prognosis.Glioma cells were randomly divided into control,SIRT7 knockdown,SIRT7 overexpression,drug treatment(10 μmol/L hydrochlorothiazide)and drug(10 μmol/L hydrochlorothiazide)+SIRT7 overexpression groups.The CCK-8 assay,cell scratch assay and Transwell assay were used to observe the effects of upregulating and downregulating SIRT7 expression on glioma cell proliferation,migration and invasion.RT-qPCR and Western blotting were employed to detect the effects of SIRT7 on the expression of neural cadherin(N-cadherin),Vimentin,E-cadherin,transforming growth factor-β(TGF-β),Ki-67,and Smad3 protein in glioma cells.Nude mouse tumor-bearing experiments were conducted to observe the effect of SIRT7 knockdown on glioma growth.Results Higher expression levels of SIRT7 gene were associated with poorer clinical prognosis(P<0.0001).SIRT7 expression levels were significantly correlated with tumor grading and 1p19q coding status(P<0.01).Compared with normal HA cells,glioma cells showed significantly increased SIRT7 expression levels(P<0.01).CCK-8 assay results indicated that,compared with control group,the proliferation activity of glioma cells in SIRT7 knockout group was significantly decreased(P<0.01),while SIRT7 overexpression group showed significantly increased proliferation activity(P<0.01).EdU assay results showed that,compared with control group,the proportion of glioma cells in the proliferative stage was significantly decreased in SIRT7 knockdown group(P<0.01),and significantly increased in SIRT7 overexpression group(P<0.01).Western blotting results revealed that,compared with control group,the protein expression levels of TGF-β,Smad3,N-cadherin and Vimentin were significantly decreased in SIRT7 knockdown group(P<0.01),while the expression level of E-cadherin protein was significantly increased(P<0.05).SIRT7 overexpression group showed significantly increased protein expression levels of TGF-β,Smad3,N-cadherin and Vimentin(P<0.05),and a significantly decrease in E-cadherin protein expression level(P<0.05).Scratch assay results indicated that,compared with control group,the migration ability of cells in SIRT7 knockdown group and drug group was significantly decreased(P<0.01),and SIRT7 overexpression group showed significantly increased cell migration ability(P<0.05).Compared with drug group,drug+SIRT7 overexpression group exhibited significantly increased cell migration ability(P<0.01).Transwell assay results showed that,compared with control group,the migration and invasion abilities of cells in SIRT7 knockdown group and drug group were significantly decreased(P<0.01),and SIRT7 overexpression group exhibited significantly increased migration and invasion abilities(P<0.01).Compared with drug group,drug+SIRT7 overexpression group showed significantly increased migration and invasion abilities(P<0.01).Nude mouse tumor-bearing assay results indicated that the volume and weight of glioma in SIRT7 knockdown group were significantly reduced compared with control group(P<0.01).Conclusions Glioma patients with high SIRT7 expression have poorer clinical prognosis.SIRT7 can regulate the TGF-β/Smad3 pathway to mediate EMT,promoting the proliferation and migration of glioma cells.SIRT7 knockdown can inhibit the growth of transplanted gliomas in nude mice.
7.Protective effect of secreted secretomes derived from three-dimensional cultured mesenchymal stem cells on mouse hippocampal neurons with seawater immersion and trauma
Xiao-Juan LAN ; Sheng-Qiang XIE ; Bing-Xian WANG ; Wen-Jing MA ; Qiao-Mi CHEN ; Gang CHENG ; Wen-Ying LYU ; Jian-Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2025;50(3):324-331
Objective To investigate the protective effects of secretomes released by three-dimensional cultured mesenchymal stem cells(MSCs)on neurons subjected to seawater immersion(SW)and stretch injury(SI),and to provide new insights into neuronal repair following SW combined with traumatic brain injury(TBI).Methods MSCs were cultured using the hanging drop method,and the conditioned medium(CM)containing MSCs secretomes was collected.A cellular model combining SW with SI was established using mouse hippocampal neuronal cells(HT22 cells).HT22 cells were randomly assigned to five groups:control,SI,SI+SW,SI+CM,and SI+SW+CM groups.Cell viability was assessed using the CCK-8 assay,apoptosis rate was measured by flow cytometry,cell migration ability was evaluated by scratch assay,and the expression levels of apoptosis-related proteins Bcl-2 and Bcl-2-associated protein(Bax),and ferroptosis-related proteins long-chain acyl-CoA synthetase 4(ACSL4)and cyclooxygenase-2(COX-2)were detected by Western blotting.Results Immersion in 15%seawater for 12 h significantly decreased HT22 cell viability(P<0.05).The CCK-8 assay indicated that cell viability in both the SI and SI+SW groups was significantly lower than that in control group after 12 h of treatment(P<0.05).Treatment with CM containing MSCs secretomes significantly increased cell viability in SI+CM group compared to SI group(P<0.0001),and in SI+SW+CM group compared to SI+SW group(P<0.001).Flow cytometry results revealed that the apoptosis rate in SI and SI+SW groups was significantly higher than that in control group(P<0.05 or P<0.001),while in SI+CM group was lower than that in SI group(P<0.05),and in SI+SW+CM group was lower than that in SI+SW group(P<0.05).Western blotting showed that compared to control group,SI and SI+SW groups exhibited reduced Bcl-2 expression level(P<0.01 or P<0.0001)and increased expression levels of Bax,ACSL4,and COX-2(P<0.01 or P<0.0001).Compared to SI group,the SI+CM group displayed increased Bcl-2 expression level(P<0.05)and decreased expression levels of Bax,ACSL4,and COX-2(P<0.05).Compared to SI+SW group,SI+SW+CM group exhibited increased Bcl-2 expression level(P<0.01)and decreased expression levels of Bax,ACSL4,and COX-2(P<0.01 or P<0.001).Scratch assay results demonstrated that at both 12 h and 24 h,the cell migration rate in SI and SI+SW groups was significantly lower than that in control group(P<0.01 or P<0.0001),while the migration rate in SI+CM group was significantly higher than that in SI group(P<0.0001 or P<0.01),and the migration rate in SI+SW+CM group was significantly higher than that in SI+SW group(P<0.0001).Conclusion Secretomes derived from MSCs cultured using the hanging drop method can alleviate neuronal damage caused by SW and TBI,potentially offering a therapeutic approach for SW combined with TBI.
8.Establishment and evaluation of a lipopolysaccharide-induced acute respiratory distress syndrome model in minipigs
Chuang-Ye WANG ; Ran WANG ; Jian ZHANG ; Ling-Xiao QIU ; Bin QING ; Heng YOU ; Jin-Cheng LIU ; Bin WANG ; Nan-Bo WANG ; Jia-Yu LI ; Xing LIU ; Shuang WANG ; Jin HU ; Jian WEN ; Quan LI ; Xiao-Ou HUANG ; Kun ZHAO ; Shuang-Lin LIU ; Gang LIU ; Mei-Ju WANG ; Qing XIANG ; Hong-Mei WU ; Xiao-Rong SUN ; Tao GU ; Dong ZHANG ; Qi LI ; Zhi XU
Medical Journal of Chinese People's Liberation Army 2025;50(9):1154-1161
Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.
9.Effects of Yiqi Juanbi Formula on chondrocyte pyroptosis in collagen-induced arthritic rats via NF-κB/NLRP3/Caspase-1 signaling pathway
Xin-yu CUI ; Hao-lin LI ; Wei-qing LI ; Hui-qin KANG ; Wei-gang CHENG ; Pei-xin HE ; Cai-hong YANG ; Ping CHEN ; Hai-dong WANG
Chinese Traditional Patent Medicine 2025;47(9):2880-2887
AIM To investigate the effects of Yiqi Juanbi Formula on chondrocyte pyroptosis in rat models of collagen-induced arthritis(CIA).METHODS Fifty rats were subcutaneously injected at the tail base with an emulsion containing equal volumes of bovine type Ⅱ collagen and incomplete Freund's adjuvant(IFA)to establish the CIA models.These rats were then randomly assigned to the model group,the methotrexate group(0.35 mg/kg),and the low-dose,medium-dose,and high-dose Yiqi Juanbi Formula groups(9.4,18.7,37.4 g/kg),in contrast to the ten intact rats serving in the normal control group.Following four weeks of intragastric administration,the rats had their general conditions observed;their joint swelling and arthritis indices measured;their ankle joint pathology assessed by HE staining;their serum levels of IL-1β,IL-18 and TNF-ɑ detected by ELISA;their mRNA expressions of NLRP3,Caspase-1,GSDMD,IL-1β,IL-18 and TNF-ɑ in ankle cartilage quantified by RT-qPCR;their protein expressions of NF-κB,NLRP3 and Caspase-1 in ankle cartilage analyzed by Western blot;and their NLRP3 and GSDMD positive expressions in ankle cartilage examined by immunohistochemistry.RESULTS Compared to the control group,the model group showed significantly increased joint swelling and arthritis indices(P<0.01);elevated serum levels of IL-1 β,IL-18 and TNF-ɑ(P<0.01);pathological changes including cartilage surface defects,reduced cell count,altered cellular morphology,irregular cell arrangement,and significant inflammatory cell infiltration in synovial tissue;upregulated mRNA expressions of NF-κB,NLRP3,Caspase-1,GSDMD,IL-1β,IL-18 and TNF-ɑ(P<0.01)and increased protein expressions of NF-κB,NLRP3 and Caspase-1(P<0.01)in ankle cartilage;enhanced positive expressions of NLRP3 and GSDMD in ankle cartilage(P<0.01).Compared to the model group,the groups intervened with methotrexate or medium-or high-dose Yiqi Juanbi Formula exhibited reduced joint swelling and arthritis indices(P<0.01);alleviated pathological damage in ankle joints;decreased serum levels of IL-1β,IL-18 and TNF-ɑ(P<0.01);downregulated mRNA expressions of NF-κB,NLRP3,Caspase-1,GSDMD,IL-1β,IL-18 and TNF-ɑ(P<0.05,P<0.01),and reduced protein expressions of NF-κB,NLRP3 and Caspase-1(P<0.05,P<0.01)in ankle cartilage;and diminished positive expressions of NLRP3 and GSDMD in ankle cartilage(P<0.01).CONCLUSION Yiqi Juanbi Formula alleviates inflammation in CIA rats,potentially by inhibiting the activation of the NF-κB/NLRP3/Caspase-1 signaling pathway,thereby suppressing chondrocyte pyroptosis.
10.Role and mechanism of RNA m6A methyltransferase WTAP in epithelial-mesenchymal transition of glioblastoma cells
Cong-Wei LIU ; Wen-Ying LYU ; Shu-Wei WANG ; Sheng-Qiang XIE ; CHENG-GANG ; Jian-Ning ZHANG
Medical Journal of Chinese People's Liberation Army 2025;50(11):1433-1443
Objective To explore the role and mechanism of RNA m6A methyltransferase Wilms'tumor 1-associated protein(WTAP)in epithelial-mesenchymal transition(EMT)of glioblastoma cells and its association with transcription factor JUNB.Methods(1)Based on the Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases,the expression levels of transforming growth factor β(TGF-β),WTAP,and JUNB in glioblastoma multiforme(GBM)and normal brain tissues were analyzed,as well as their diagnostic and prognostic values for GBM.The correlation of TGF-β,WTAP,and JUNB with gliomas of different WHO grades was analyzed using the Chinese Glioma Genome Atlas(CGGA)database.Spearman correlation analysis was performed to assess the correlation between TGF-β and m6A methyltransferases.(2)An EMT model was established in human astrocytoma U87-MG cells through TGF-β1 induction.qRT-PCR and Western blotting were employed to detect the expression levels of WTAP,JUNB,matrix metalloproteinase 2(MMP2),and N-cadherin.The migration capacity of U87-MG cells was evaluated by wound-healing and Transwell assays.An m6A RNA methylation quantification kit(colorimetric)was used to detect RNA m6A methylation modification levels.A stable cell line with low expression of WTAP was constructed to investigate the effects of WTAP knockdown on the migration ability of U87-MG cells,as well as the expression of JUNB.(3)A protein-protein interaction network was constructed using STRING database and GeneMANIA database,followed by gene ontology(GO)and KEGG pathway enrichment analyses to explore the biological processes,molecular functions,cellular components,and signaling pathways potentially involved in TGF-β/WTAP/JUNB.Gene set enrichment analysis(GSEA)was performed on JUNB-related genes to investigate their potential downstream signaling pathways.Results(1)The expression levels of TGF-β,WTAP,and JUNB were significantly higher in GBM(P<0.001),positively correlated with WHO grades of glioma(P<0.001).Glioma patients with high expression of all three genes had shorter overall and disease-free survival(P<0.001).Spearman analysis showed that the expression of TGF-β in GBM was positively correlated with WTAP(r=0.175,P=0.023),but no significant correlation with other m6A methyltransferases(P>0.05).(2)After TGF-β1 treatment,the level of m6A methylation modification of total RNA in U87-MG cells significantly increased(P<0.001).Wound-healing assay and Transwell assay results showed that the migration ability of U87-MG cells was significantly increased after TGF-β1 treatment(P<0.01),while WTAP knockdown significantly reduced the migration ability of U87-MG cells(P<0.01).qRT-PCR and Western blotting results showed that the mRNA and protein expression levels of WTAP,N-Cadherin,MMP2,and JUNB in U87-MG cells were significantly increased after 48 h of TGF-β1 induction(P<0.001),while WTAP knockdown significantly reduced the mRNA and protein expression of JUNB(P<0.001).(3)The TGF-β/WTAP/JUNB-related protein-protein interaction network was constructed,which was primary involved in mRNA modification and EMT.GSEA results showed that JUNB-related signaling pathways were closely associated with glioma malignant progression.Conclusions TGF-β,WTAP,and JUNB are all associated with GBM malignant progression and poor patient prognosis.TGF-β may enhance total RNA m6A modification by promoting the expression of m6A methyltransferase WTAP,and WTAP subsquentaly upregulates transcription factor JUNB,thereby promoting EMT and malignant progression of GBM.

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