1.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.
2.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.
3.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.
4.Evaluation of diagnostic efficacy and severity of necrotizing enterocolitis in premature infants by intestinal regional oxygen saturation and fecal calprotectin
Lingyu FANG ; Jiangbin CHEN ; Zhiyong LIU ; He WANG ; Dongmei CHEN
Chinese Pediatric Emergency Medicine 2024;31(1):35-40
Objective:To evaluate the diagnostic value of intestinal regional oxygen saturation(rSO 2)and fecal calprotectin in the occurrence and severity of necrotizing enterocolitis(NEC)in premature infants. Methods:A prospective observational study was conducted among premature infants admitted to Quanzhou Children's Hospital from October 2019 to December 2022. Intestinal rSO 2 was monitored within two hours of diagnosis of NEC,and fecal calprotectin was measured. Results:A total of 60 patients were included, including 30 cases with NEC and 30 cases without NEC, 14 cases of medical NEC, 16 cases of surgical NEC, and eight infants died due to NEC. Infants with NEC had lower intestinal rSO 2 [49(30,60)% vs. 66(60,69)%] and higher calprotectin levels [479(297,886)μg/g vs. 203(113,275)μg/g] than those in infants without NEC ( P<0.01). The levels of intestinal rSO 2 were lower in surgical NEC than those in medical NEC,and were lower in the death group than that in the survival group ( P<0.01),but no similar difference was found in the levels of calprotectin. ROC curve analysis showed that intestinal rSO 2 combined with calprotectin had a sensitivity of 73%,a specificity of 100%,and the largest area under curve of 0.91 in the diagnosis of NEC. Intestinal rSO 2 had an optimal cut-off value of 31% in predicting death in infants with NEC,with a sensitivity of 100%,a specificity of 95%,and an area under curve of 0.99. Conclusion:Intestinal rSO 2 and fecal calprotectin can effectively identify the presence of NEC,and their combined detection can improve the diagnostic efficiency. Intestinal rSO 2 is a good predictor of the severity of NEC,but not fecal calprotectin.
5.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.
6.Construction of a predictive model for postoperative subsyndromal delirium in elderly patients undergoing abdominal surgery
Shunli CHEN ; Yan LIU ; Jing CHEN ; Lingyu TANG ; Ru SONG ; Luyao CAI ; Silin ZHENG
Chinese Journal of Nursing 2024;59(16):2001-2008
Objective To analyze the influencing factors of postoperative subsyndromal delirium in elderly patients undergoing abdominal surgery,and to develop a nomogram risk prediction model.Methods A convenience sampling method was used to select 497 elderly patients who underwent abdominal surgery in a tertiary hospital in Sichuan Province from February to October 2022.The risk prediction model and nomograms model were constructed using Logistic regression analysis and R software.The area under the subject's working characteristic curve(AUC)and Hosmer-Lemshow test were used to evaluate the discrimination and calibration of the model.Results The results of Logistic regression showed that age(OR=1.066),functional activities(OR=1.143),patient controlled intravenous analgesia(OR=5.811),transanal drainage tube(OR=2.276)and postoperative blood transfusion(OR=4.322)were independent influences on the occurrence of postoperative subsyndromal delirium.The p-values of the prediction model in both the training and validation sets were greater than 0.05;the area under the ROC curves were 0.734 and 0.691;the model was presented in the form of nomogram.Conclusion The prediction model developed in this study has good discrimination and accuracy.It can be used to assist clinical staff in identifying patients at high risk of developing postoperative subsyndromal delirium and provide the reference for developing preventive and intervention measures.
7.Application progress of Satir communication model in nursing from the perspective of family support
Yimeng CHEN ; Yinning GUO ; Hanfei ZHU ; Kang ZHAO ; Ting XU ; Lidong HUANG ; Lingyu DING ; Jieman HU ; Qin XU
Chinese Journal of Nursing 2024;59(19):2413-2419
As a type of experiential psychotherapy,Satir communication model can help the individual system and the family system achieve a state from dysfunction to healthy function,which can enrich the intervention connotation of family support and provide a new direction for the realization of full-life circle care.This paper aims to introduce the concept,core elements,common treatment techniques,application and effects,current challenges and relevant suggestions of Satir communication model in the nursing field from the perspective of family support,in order to provide references for the localization development and clinical integration of Satir communication model in the field of nursing in China.
8.Interaction Between Bruceoside B and Intestinal Flora and Its Inhibitory Effect on Human Lung Cancer A549 Cells
Lingyu SHI ; Wenmin WANG ; Yulin FENG ; Shilin YANG ; Yang WAN ; Daofeng CHEN ; Quan WEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(13):160-166
ObjectiveTo explore the interaction between bruceoside B and gut microbiota and the inhibitory activity of its metabolites on human lung cancer A549 cells, and to explore the value of bruceoside B in the treatment of non-small cell lung cancer(NSCLC). MethodBruceoside B was co-incubated with the human gut microbiota under anoxic conditions in vitro, and ultra high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to analyze the metabolic transformation products. Cell counting kit-8(CCK-8) assay was performed to determine the effects of bruceoside B and its metabolites on the proliferation of human lung cancer A549 cells and the half inhibitory concentration(IC50) was calculated. Five healthy male rats were gavaged with bruceoside B(2 mg·kg-1) for 7 days after adaptive feeding. The feces of rats were collected before and after administration. 16S rRNA sequencing was used to assess gut microbiota. ResultBruceoside B was mainly metabolized to brusatol by human gut microbiota, the IC50 of bruceoside B and the conversion product to A549 cells were 1 755.50, 19.57 μmol·L-1, respectively, and the conversion product had a better activity at inhibiting A549 cells proliferation than bruceoside B. Additionally, The results of intestinal flora analysis showed no significant differences in α diversity and β diversity of gut microbiota after administration. In terms of species abundance, at the phylum level, bruceoside B decreased the relative abundance of Actinobacteriota and Proteobacteria, increased the relative abundance of Firmicutes, Patescibacteria and Cyanobacteria. At the genus level, bruceoside B decreased the relative abundance of Staphylococcus, Aerococcus and Psychrobacter, increased the relative abundance of Romboutsia, Lactobacillus, Clostridium sensu stricto 1, Norank-f-norank-o-Clostridia-UCG-014, Turicibacter, Allobaculum and Candidatus Saccharimonas. The results of functional prediction showed that the gut microbiota functional compositions were relatively stable. ConclusionBruceoside B can be deglycosylated by intestinal flora and converted into brusatol, with a significant increase in antitumor activity. The administration of bruceoside B will not cause significant changes in the structure and function of the intestinal flora, resulting in intestinal microecological balance disorders, and the administration appears to be beneficial to the intestinal flora of NSCLC patients.
9.Research and application of digital technique combined with 3D printing guide plate in surgery of maxillofacial fracture
Xiaojie GAO ; Bing LIU ; Bing LI ; Bo WEI ; Lingyu SHEN ; Peng CHEN
China Medical Equipment 2024;21(5):98-101,106
Objective:To explore the applied therapeutic effect of digital surgical technique combined with three dimensional(3D)printing guide plate in surgery of maxillofacial fracture.Methods:A total of forty patients with maxillofacial fracture who admitted to the Department of Stomatology at the First Medical Center of Chinese PLA General Hospital from June 2019 to June 2023 were selected.They were divided into an observation group(18 cases)and a control group(22 cases)based on different surgical methods.The observation group used digital technique combined with 3D printing technique to conduct surgery by auxiliary method,and the control group used routine open reduction and internal fixation surgery for maxillofacial fracture.The surgical time,bleeding volume,complications and satisfaction evaluation between two groups of patients were observed and compared.Results:All patients obtained clinical cure after open reduction and internal fixation surgery,and the surgical time(2.01±0.52)h and bleeding volume(123.89±67.31)mL during surgery of observation group were significantly lower than those of control group(U=62.000,26.500,P<0.05),respectively.The incidence of the complications in the observation group was significantly lower than that in the control group,and the difference was statistically significant(x2=4.675,P<0.05),and there were no statistically significant difference in infection and limited incision opening between the two groups(P>0.05).In the satisfaction evaluation,there was a significant difference in appearance satisfaction scores between the observation group(8.89±0.758)and the control group(7.73±0.827)(U=66.000,P<0.05),while there were no statistically significant differences in chewing and language aspects between two groups(P>0.05).Conclusion:The combination of digital technique and 3D printing guide plate has favorable clinical guidance effect in maxillofacial fracture surgery,which can significantly improve surgical efficacy,and reduce the incidence of complications,and obtain higher patient satisfaction.It is worth to be applied in clinical practice.
10.Impacts of X-rays at varying doses on liver injury and oxidative stress in mice
Mingfang LI ; Lingyu ZHANG ; Lina CAI ; Sufen ZHANG ; Yashi CAI ; Yuhua YANG ; Huifeng CHEN ; Jianming ZOU ; Weixu HUANG
Chinese Journal of Radiological Medicine and Protection 2024;44(9):734-740
Objective:To investigate the changes in liver injury and oxidative-antioxidant level in mice exposed to X-rays at varying doses.Methods:Fifty-four 8-week-old male C57BL/6J mice were divided into three groups, namely the control, 2 Gy irradiation, and 4 Gy irradiation groups. Then, each of the groups was further divided by days post-irradiation (i.e., 1, 3, and 7 d), and so nine sub-groups ( n = 6). After irradiation was performed as planned, all the mice were dissected and weighed, and their liver indexes were calculated to determine any histopathological changes in the liver. The peripheral blood cell count and the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were detected. Furthermore, spectrophotometry was also used to determine the superoxide dismutase (SOD) activity, the malondialdehyde (MDA) concentration, and the reduced glutathione (GSH) concentration in liver tissues. Results:Compared to the control group, mice undergoing irradiation exhibited a significant reduction in body weight ( F = 84.03, 27.11, 25.50, P < 0.001), but significantly increased liver indexes ( F = 28.40, 17.75, P <0.001) at 1, 3, and 7 d post-irradiation. Pathological observations of these mice revealed liver injury, which proved related to dose and time course. The counts of leukocytes, neutrophils, and lymphocytes in peripheral blood decreased significantly ( F = 8.42-22.91, P < 0.05), trending downward with an increase in the radiation dose. For mice in the 4 Gy irradiation group, their AST and ALT levels increased significantly at 1 d post-irradiation ( H = 7.24, 7.82, P < 0.05), and their ALP levels rose notably at 1 and 3 d post-irradiation ( F = 11.86, 9.75, P < 0.05). Furthermore, their MDA and SOD levels initially rose and then dropped but their GSH levels exhibited an opposite trend at 1, 3, and 7 d post-irradiation. There was a positive correlation between their MDA levels in the liver and the degree of damage to histopathological lesions at 1, 3, and 7 d post-irradiation ( r = 0.30, P < 0.001). Conclusions:A model for radiation-induced liver injury of mice was preliminarily established in this study. It can be concluded that X-rays at varying doses affect the severity of liver injury, pathological grade, peripheral blood cell count, liver function index, and liver oxidative and antioxidant levels of mice, presenting a certain relationship between dose and time course effects.

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