1.MiRSNP in MGMT 3′ untranslated region regulates low-dose radiation-induced senescence in lung cells
Lingyu ZHANG ; Yashi CAI ; Huixian LI ; Min ZHANG ; Changyong WEN ; Weixu HUANG ; Huifeng CHEN ; Jianming ZOU
China Occupational Medicine 2025;52(1):25-32
Objective To investigate the role of the O-6-methylguanine-DNA methyltransferase (MGMT) gene-3′ untranslated region (UTR) microRNA-associated single nucleotide polymorphism (miRSNP) (rs7896488 G>A) in affecting miR-4297-targeted modulation of MGMT in senescence of lung cells with polymorphic genotypes induced by fractionated low dose ionizing radiation (LDIR). Methods i) MiRSNPs were predicted and screened using bioinformatics, and DNA from two types of lung cells, A549 cells and human bronchial epithelioid cells (HBE cells), was extracted for target gene sequencing. After co-transfection of pGL3c-MGMT-3′UTR-rs7896488 G>A reporter gene recombinant plasmid, pRL-TK Vector with micrON mimic NC #22 or micrON hsa-miR-4297 mimic (set up as the mimic NC group and the miR-4297 mimic group) in these two types of lung cells, dual luciferase reporter gene assay was performed. The relative expression of MGMT mRNA was detected by real-time fluorescence quantitative polymerase chain reaction, and the relative expression of MGMT protein was detected by Western blotting. ii) These two types of lung cells were randomly divided into the control group and irradiation group, which received either 0 or 100 mGy X-rays irradiation seven times. After irradiation, the cells were transfected with either micrON mimic NC #22 or micrON hsa-miR-4297 mimic, resulting in mimic NC + control group, miR-4297 mimic + control group, mimic NC + irradiation group, and miR-4297 mimic + irradiation group. Cells were collected for senescence-associated-β-galactosidase (SA-β-Gal) staining, and the relative expression of matrix metalloproteinase-9 (MMP-9) and chemokine (C-X-C motif) ligand-1 (CXCL-1) proteins was detected via Western blotting. Results i) The rs7896488 G>A was the miRSNP located in the conserved binding region targeted by miR-4297 in the MGMT gene 3′UTR. A549 cells were the rs7896488 GG wild-type homozygous genotype, while HBE cells were the rs7896488 GA heterozygous mutant genotype. In the miR-4297 mimic group, A549 and HBE cells carrying the rs7896488 G allele showed significantly lower dual-luciferase activity compared with that in the mimic NC group (both P<0.01). However, there was no significant difference in dual-luciferase activity between the two groups in both A549 and HBE cells carrying the rs7896488 A allele (both P>0.05). The relative expression levels of MGMT mRNA and MGMT protein of A549 cells in the miR-4297 mimic group were lower than those in the mimic NC group (both P<0.05). However, there was no significant difference in MGMT mRNA and MGMT protein of HBE cells between these two groups (both P>0.05). ii) The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of A549 cells in the miR-4297 mimic+irradiation group were higher than those in the mimic NC + control group, the miR-4297 mimic + control group, and the mimic NC + irradiation group (all P<0.05). The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of HBE cells in the miR-4297 mimic + irradiation group were higher than those in the mimic NC + control group and the miR-4297 mimic + control group (all P<0.05), while there was no significant difference compared with those in the mimic NC + irradiation group (all P>0.05). Conclusion MGMT-3′UTR-miRSNP rs7896488 G>A plays a role in LDIR-induced senescence of lung cells with different polymorphic genotypes by affecting miR-4297-targeted regulation of MGMT.
2.Effects of fractionated low-dose ionizing radiation on differentially expressed genes in ferroptosis of human bronchial epithelial cells
Min ZHANG ; Lingyu ZHANG ; Yashi CAI ; Huixian LI ; Yanting CHEN ; Guanyou CHEN ; Xin LAN ; Changyong WEN ; Weixu HUANG ; Jianming ZOU ; Huifeng CHEN
Chinese Journal of Radiological Health 2025;34(3):310-317
Objective To investigate the effects of fractionated low-dose ionizing radiation (LDIR) on the ferroptosis in human bronchial epithelial (HBE) cells as well as the associated differentially expressed genes (DEGs), biological processes, and signaling pathways. Methods HBE cells were exposed to different single doses of X-ray irradiation (0, 25, 50, 75, and 100 mGy) for 24, 48, and 72 h, respectively. The change in cell viability was detected by MTT assay. Cells were irradiated with 0, 25, 50, and 100 mGy X-rays 5 times, with 48 h between each irradiation and a dose rate of 50 mGy/min. Cells were harvested 24 h after irradiation for the measurement of the expression of ferroptosis-related genes SLC7A11 and GPX4 at the mRNA and protein levels, cellular iron content, and the expression of FTH1 and FTL mRNAs. High-throughput sequencing was used to screen for the DEGs in each dose group, followed by Gene Ontology-Biological Process (GO-BP) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and Gene Set Enrichment Analysis (GSEA). Results Compared with the control group, single-dose LDIR significantly increased cell proliferation at 75 mGy after 24 h (P < 0.05), at 50, 75, and 100 mGy after 48 h (P < 0.05), and at 75 and 100 mGy after 72 h (P < 0.05). Compared with the control group, at the end of the fifth fractionated LDIR, SLC7A11 and GPX4 mRNAs decreased at all doses (P < 0.05), SLC7A11 protein decreased at all doses, GPX4 protein decreased at 25 and 100 mGy, iron content increased at all doses, and FTH1 and FTL mRNAs decreased at all doses (P< 0.05). Sequencing analysis identified 248, 30, and 291 DEGs and 10, 2, and 9 ferroptosis-associated genes at the three doses compared to the control. Gene Ontology-Biological Process analysis showed that DEGs were mainly enriched in biological processes such as response to lipids, cell death, and response to unfolded proteins. Kyoto Encyclopedia of Genes and Genomes analysis showed that DEGs were mainly enriched in the JAK-STAT signaling pathway, lipids and atherosclerosis, ferroptosis, protein processing in the endoplasmic reticulum, and FoxO signaling pathway. Gene set enrichment analysis showed that DEGs were mainly enriched in ferroptosis, fatty acid degradation, and glutathione metabolism. Conclusion Fractionated low-dose radiation induced ferroptosis in HBE cells, and DEGs were predominantly enriched in biological processes and signaling pathways related to inflammation, ferroptosis, and endoplasmic reticulum stress.
3.Association between long-term exposure to low-dose ionizing radiation and metabolic syndrome among medical radiologists
Changyong WEN ; Xiaoman ZHOU ; Xiaolian LIU ; Yiqing LIAN ; Weizhen GUO ; Yanting CHEN ; Xin LAN ; Mingfang LI ; Sufen ZHANG ; Weixu HUANG ; Jianming ZOU ; Huifeng CHEN
Journal of Environmental and Occupational Medicine 2025;42(10):1209-1215
Background In recent years, the increasingly widespread application of nuclear and medical radiation technologies has resulted in a large number of occupational populations exposed to low-dose ionizing radiation (LDIR). At present, there is no consistent conclusion on the effects of long-term exposure to LDIR on the metabolic health of the occupational population. Objective To explore the association between long-term exposure to LDIR and metabolic syndrome (MetS) among medical radiologists. Methods A cross-sectional study was conducted to enroll
4.Effect of melatonin-modified PEEK implant on osteointegration in osteoporotic rats
Guisheng LUO ; Teng GU ; Junjun LI ; Penglai WANG ; Cheng ZHANG ; Changyong YUAN
STOMATOLOGY 2024;44(10):734-741
Objective To explore the effect of melatonin(MT)-modified PEEK implant assisted by polydopamine(PDA)coating on osteointegration in osteoporotic rats.Methods MT was adhered to PEEK implants with PDA coating as carrier.The physicochemical properties of the materials were analyzed by SEM image,water contact angle,FTIR and protein adsorption experiment.OVX-rBMSCs were inoculated on the surface of PEEK sheet and cultured.The cytoskeleton was stained and cell adhesion morphology was observed.Cell proliferation activity was evaluated by CCK-8 assay;key enzyme activities for osteogenic differentiation were analyzed by ALP stai-ning,and expression levels of osteoblast-related genes COL-1,Runx2,OPN,OCN,BMP-2 and ALP were detected by quantitative real-time PCR.In addition,implants were implanted into the femur of osteoporotic rats and bone volume on the implant surface was de-tected and quantified by Micro-CT.Results MT was successfully loaded on PEEK;the cell adhesion was better,and the proliferation activity and osteogenic differentiation ability were significantly higher than those of control group(P<0.01).In the rat osteoporosis mod-el,there was more new bone formation around the modified PEEK implant(P<0.01).Conclusion MT-modified PEEK implants have excellent biocompatibility and improve osteointegration in an osteoporotic environment.
5.Differential mRNA expression in fractional low-dose radiation-induced senescence of HBE cells
Lingyu ZHANG ; Weixu HUANG ; Yashi CAI ; Huixian LI ; Min ZHANG ; Changyong WEN ; Ping YANG ; Jianming ZOU ; Huifeng CHEN
Chinese Journal of Radiological Health 2024;33(2):116-122
Objective To explore the differentially expressed mRNAs and related biological processes and pathways in fractional low-dose ionizing radiation (LDIR)-induced senescence of normal human bronchial epithelial (HBE) cells by high-throughput mRNA sequencing and bioinformatics techniques. Methods Senescence-associated β-galactosidase staining and senescence-associated secretion phenotype gene mRNA and protein expression levels were measured at 24 and 48 h after irradiating HBE cells 7 times at doses of 0, 50, 100, and 200 mGy, respectively. The differentially expressed genes were screened by high-throughput sequencing for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Results The senescence-positive area of fractional low-dose irradiated HBE cells increased in a dose-dependent manner (P < 0.05). The mRNA levels and protein expression of transforming growth factor-β1(TGF-β1) and matrix metalloproteinase-9(MMP-9) genes were increased in the 100 mGy × 7 and 200 mGy × 7 groups at 24 and 48 h after the end of irradiation compared with the control group. High-throughput sequencing showed that there were 882, 475, and 1205 differentially expressed mRNAs in each dose group compared with the control group. GO analysis showed that the differentially expressed mRNAs in each dose group were mainly enriched in biological processes such as cell cycle regulation, regulation of nitrogen compound metabolic process, regulation of cell division and response to stimulus. KEGG analysis showed that the differentially expressed mRNAs were mainly enriched in the pathways of cell cycle, cell senescence, and ferroptosis. Conclusion Fractional LDIR induced senescence in HBE cells, and differentially expressed mRNA-associated biological processes and pathways in senescent cells are related to cell cycle and cell senescence.
6.Effects of fractionated low-dose ionizing radiation in the induction of EA.hy926 cell senescence
Yashi CAI ; Weixu HUANG ; Lingyu ZHANG ; Min ZHANG ; Huixian LI ; Changyong WEN ; Zhini HE ; Jianming ZOU ; Huifeng CHEN
Chinese Journal of Radiological Health 2024;33(1):13-20
Objective To investigate the mechanism of fractionated low-dose ionizing radiation (LDIR) in the induction of EA.hy926 cell senescence. Methods EA.hy926 cells were irradiated with X-ray at 0, 50, 100, and 200 mGy × 4, respectively, and cultured for 24, 48, and 72 h. Several indicators were measured, including the levels of cellular senescence-associated β-galactosidase (SA-β-gal) staining, mRNA levels of senescence-associated cell cycle protein-dependent kinase inhibitor genes CDKN1A and CDKN2A, reactive oxygen species (ROS), total antioxidant capacity (T-AOC), and phosphorylated H2A histone family member X (γ-H2AX). Results After 4 fractionated LDIR, compared with the control group, the treatment groups showed increased nucleus area, blurred cell edge, and increased SA-β-gal positive area (P < 0.05) at 24, 48 and 72 h. After 4 fractionated LDIR, the mRNA level of CDKN1A increased in the 100 and 200 mGy × 4 groups at 24 and 48 h (P < 0.05), and CDKN2A mRNA level increased in the 100 and 200 mGy × 4 groups at 48 and 72 h (P < 0.05). The fluorescence intensity of ROS increased in treatment groups at 24, 48, and 72 h after 4 fractionated LDIR (P < 0.05). After 4 fractionated LDIR, the T-AOC level increased in the 100 and 200 mGy × 4 groups at 24 h (P < 0.05), and T-AOC level increased in all treatment groups at 48 and 72 h (P < 0.05). After 4 fractionated LDIR, γ-H2AX fluorescence intensity increased in all treatment groups at 24 h (P < 0.05), and the fluorescence intensity increased in the 100 and 200 mGy × 4 groups at 48 and 72 h (P < 0.05). Conclusion Fractionated LDIR can induce cellular senescence in EA.hy926 cells by impacting the cellular oxidation-antioxidation and oxidative damage levels, and the effects were relatively evident at 100 and 200 mGy.
7.Repair of congenital earlobe defect using retroauricular three flaps
Changyong CHEN ; Ling ZHANG ; Qinghua YANG
Chinese Journal of Plastic Surgery 2023;39(1):65-70
Objective:To investigate the method and effect of retroauricular three flaps for repair of congenital earlobe defect.Methods:From March 2019 to December 2021, the data of patients with congenital earlobe defect treated with retroauricular three-flap method in Beijing Anzhen Hospital, Capital Medical University and Plastic Surgery Hospital, Chinese Academy of Medical Sciences were retrospectively analyzed. The specific method was to design three flaps in the posterior auricular region, A, B, C respectively. The area of flap A was larger 10%-20% than the defect area, and the flap A was rotated in the direction of the anterior and inferior earlobe. The flap B was designed as a rotation flap to repair the secondary defect after flap transplantation. The flap C was designed as a sliding flap to reduce the tension of the wound in the retroauricular region. Postoperative observation included flap blood flow, complications of infection, hematoma, and so on, and scar after wound healing. A questionnaire was used to investigate the satisfaction of patients or guardians, with scores from 1 to 4 indicating dissatisfaction, general satisfaction, satisfaction, and very satisfaction respectively.Results:A total of 10 patients were included, including 3 males and 7 females. Aged from 7-18 years old, with an average of 11 years old. All cases were unilateral lobe defects. Postoperative infection occurred in 1 case and small local hematoma in 1 case. All cases were cured after dressing change without flap necrosis and other complications. After 3 to 6 months of follow-up, there was no flap necrosis, the color, shape and plumpness of the repaired earlobe were good, and the scar behind the ear was concealed. The satisfaction survey showed that 8 cases had great satisfaction and 2 cases had satisfaction.Conclusion:Partial or total defects of earlobe can be repaired by retroauricular three flaps. The reconstructed earlobe has natural shape, hidden scar and satisfactory clinical effect.
8.Repair of congenital earlobe defect using retroauricular three flaps
Changyong CHEN ; Ling ZHANG ; Qinghua YANG
Chinese Journal of Plastic Surgery 2023;39(1):65-70
Objective:To investigate the method and effect of retroauricular three flaps for repair of congenital earlobe defect.Methods:From March 2019 to December 2021, the data of patients with congenital earlobe defect treated with retroauricular three-flap method in Beijing Anzhen Hospital, Capital Medical University and Plastic Surgery Hospital, Chinese Academy of Medical Sciences were retrospectively analyzed. The specific method was to design three flaps in the posterior auricular region, A, B, C respectively. The area of flap A was larger 10%-20% than the defect area, and the flap A was rotated in the direction of the anterior and inferior earlobe. The flap B was designed as a rotation flap to repair the secondary defect after flap transplantation. The flap C was designed as a sliding flap to reduce the tension of the wound in the retroauricular region. Postoperative observation included flap blood flow, complications of infection, hematoma, and so on, and scar after wound healing. A questionnaire was used to investigate the satisfaction of patients or guardians, with scores from 1 to 4 indicating dissatisfaction, general satisfaction, satisfaction, and very satisfaction respectively.Results:A total of 10 patients were included, including 3 males and 7 females. Aged from 7-18 years old, with an average of 11 years old. All cases were unilateral lobe defects. Postoperative infection occurred in 1 case and small local hematoma in 1 case. All cases were cured after dressing change without flap necrosis and other complications. After 3 to 6 months of follow-up, there was no flap necrosis, the color, shape and plumpness of the repaired earlobe were good, and the scar behind the ear was concealed. The satisfaction survey showed that 8 cases had great satisfaction and 2 cases had satisfaction.Conclusion:Partial or total defects of earlobe can be repaired by retroauricular three flaps. The reconstructed earlobe has natural shape, hidden scar and satisfactory clinical effect.
9.Multi-link Linkage Research on Medical Humanistic Education by Integrating Narrative Medical Materials
Qiuju ZHANG ; Changyong MA ; Yang JIAO ; Bei LI ; Hui JIANG
Chinese Medical Ethics 2023;36(11):1220-1223
Taking the current situation and problems of medical humanities education as the background, the concept of narrative medicine was used to sort out and integrate various course materials of medical humanities, explore the multi-link linkage mode of medical humanities education, and organically integrate medical humanities courses scattered in medical education activities such as medical ethics, health law, doctor-patient communication, and health policy science with teaching stages, teaching resources, and teaching methods, forming vivid narrative medical materials throughout the entire process of medical humanities education, and improving teaching efficiency by sharing and optimizing resources. Through questionnaire analysis, it was found that there are many unsatisfactory aspects of medical humanities education. Further analysis of the issues focuses on the contradiction between learning willingness and time allocation, the lack of synchronization between teaching and social development, the lack of integration between courses, and insufficient innovation in teaching methods. Based on these, countermeasures were put forward to integrate narrative medical materials of medical humanities courses and build a platform for the application and communication of narrative medical materials.
10.Thought on Improving Medical Students’ Strain Capacity in the Application of Ethics in Clinical Teaching
Xiao ZHANG ; Changyong MA ; Dong WEI
Chinese Medical Ethics 2023;36(11):1291-1294
Improveing students’ strain capacity in the application of medical ethics is an important part of clinical teaching. This paper revealed the basic connotation of strain capacity in the application of medical ethics, emphasized the necessity for medical students to possess this quality, and emphatically expounded that training students in clinical teaching according to the five basic practical requirements of ethical adaptability. For practical problems, students’ ethical discrimination ability, ethical difficult problem handling ability, and comprehensive application ability of ethics and law should be improved according to practical requirements.

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