1.Construction of evaluation index system and theory-skill-humanity model for core competence of radiation specialist nurses
Yuanyuan SONG ; Qiaofang YANG ; Tiying DING ; Ruonan HAO ; Shan BAI ; Yajuan KANG
Chinese Journal of Practical Nursing 2024;40(11):839-845
Objective:To construct of evaluation index system and theory-skill-humanity (TSH) model for core competence of radiation specialist nurses, to provide reference for clinical training of radiation specialist nurses.Methods:In January 2023, the draft of the evaluation index system of core competence for radiation specialist nurses was development by literature reviews and semi-structured interviews; from February to March 2023, 20 experts were consulted with 2 rounds of Delphi consultations to determine the index system and determine indicator weights based on the importance assigned by experts to each indicator.Results:Among 20 experts, 17 were female and 3 were male, aged 35-65 (46.48 ± 8.19) years old. The effective recovery rate of the questionnaires was 100.0% (20/20) and 95.0% (19/20) in the first and second round, respectively. The authority coefficient of the experts was 0.937 in the second round, the Kendall coefficients of concordance in the first-level indexes, second-level indexes and third-level indexes were 0.448, 0.407, 0.464, respectively, the difference was statistically significant ( χ2 = 53.60, 148.81, 58.18, all P<0.001). The final evaluation index system of the core competence of radiation specialist nurses included 3 first-level indicators, 11 second-level indicators and 56 third-level indicators. Conclusions:The evaluation index system of the core competence of radiation specialist nurses constructed based on TSH model in the present study has certain feasibility and practicality, which can be used as the basis for the training and evaluation of radiation specialized nurses.
2.Human placental mesenchymal stem cells inhibit occurrence of pulmonary fibrosis by regulating transforming growth factor-beta 1/Smad3 signaling pathway
Jiawei CAO ; Shaorui DING ; Hua TIE ; Jing XUE ; Yuanyuan JIA ; Xueyun LIANG ; Feng LI
Chinese Journal of Tissue Engineering Research 2024;28(31):4970-4974
BACKGROUND:Human placental mesenchymal stem cells have been shown to be effective in inhibiting the development of pulmonary fibrosis,but the underlying mechanisms remain unclear. OBJECTIVE:To investigate the therapeutic effect and related mechanism of human placental mesenchymal stem cells on silica-induced pulmonary fibrosis in human embryonic lung fibroblasts(MRC-5). METHODS:CCK-8 assay was used to detect the effects of different mass concentrations of silica on the proliferation of MRC-5 at different time points.Immunofluorescence staining was used to screen out the best stimulating mass concentration and time of silica for subsequent experiments.MRC-5 cells were divided into blank group,silica group,and silica + human placental mesenchymal stem cell group.In the blank group,cells were not treated.In the silica group,MRC-5 cells were stimulated with 100 μg/mL silica for 48 hours.In the silica + human placental mesenchymal stem cell group,MRC-5 cells were stimulated with 100 μg/mL silica for 48 hours and then co-cultured with human placental mesenchymal stem cells for 24 hours.Immunofluorescence staining was used to detect the expression of α-smooth muscle actin and collagen type I in cells of each group.Western blot assay was used to detect the expressions of pulmonary fibrosis-related proteins and TGF-β1/Smad 3 signaling pathway-related proteins in cells of each group. RESULTS AND CONCLUSION:(1)CCK-8 assay results suggested that 100 μg/mL silica was the best mass concentration and time to stimulate MRC-5 cells for 48 hours.(2)Immunofluorescence staining results showed that the expression of α-smooth muscle actin and collagen type I in the silica + human placental mesenchymal stem cell group was significantly lower than that in the silica group.(3)Western blot assay results showed that compared with the silica group,the protein expression levels of α-smooth muscle actin,collagen type I,N-cadherin,fibronectin,transforming growth factor-β1,p-Smad3,and Smad3 in the silica + human placental mesenchymal stem cell group were decreased,and the expression of E-cadherin was increased.The difference was statistically significant(P<0.05).(4)The results showed that human placental mesenchymal stem cells had a significant therapeutic effect on silica-induced pulmonary fibrosis.Human placental mesenchymal stem cells can inhibit the development of pulmonary fibrosis by regulating transforming growth factor-β1/Smad3 signaling pathway.
3.National bloodstream infection bacterial resistance surveillance report (2022) : Gram-negative bacteria
Zhiying LIU ; Yunbo CHEN ; Jinru JI ; Chaoqun YING ; Qing YANG ; Haishen KONG ; Haifeng MAO ; Hui DING ; Pengpeng TIAN ; Jiangqin SONG ; Yongyun LIU ; Jiliang WANG ; Yan JIN ; Yuanyuan DAI ; Yizheng ZHOU ; Yan GENG ; Fenghong CHEN ; Lu WANG ; Yanyan LI ; Dan LIU ; Peng ZHANG ; Junmin CAO ; Xiaoyan LI ; Dijing SONG ; Xinhua QIANG ; Yanhong LI ; Qiuying ZHANG ; Guolin LIAO ; Ying HUANG ; Baohua ZHANG ; Liang GUO ; Aiyun LI ; Haiquan KANG ; Donghong HUANG ; Sijin MAN ; Zhuo LI ; Youdong YIN ; Kunpeng LIANG ; Haixin DONG ; Donghua LIU ; Hongyun XU ; Yinqiao DONG ; Rong XU ; Lin ZHENG ; Shuyan HU ; Jian LI ; Qiang LIU ; Liang LUAN ; Jilu SHEN ; Lixia ZHANG ; Bo QUAN ; Xiaoping YAN ; Xiaoyan QI ; Dengyan QIAO ; Weiping LIU ; Xiusan XIA ; Ling MENG ; Jinhua LIANG ; Ping SHEN ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2024;17(1):42-57
Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of national bloodstream infection Bacterial Resistant Investigation Collaborative System(BRICS)were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute(CLSI). WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:During the study period,9 035 strains of Gram-negative bacteria were collected from 51 hospitals,of which 7 895(87.4%)were Enterobacteriaceae and 1 140(12.6%)were non-fermenting bacteria. The top 5 bacterial species were Escherichia coli( n=4 510,49.9%), Klebsiella pneumoniae( n=2 340,25.9%), Pseudomonas aeruginosa( n=534,5.9%), Acinetobacter baumannii complex( n=405,4.5%)and Enterobacter cloacae( n=327,3.6%). The ESBLs-producing rates in Escherichia coli, Klebsiella pneumoniae and Proteus spp. were 47.1%(2 095/4 452),21.0%(427/2 033)and 41.1%(58/141),respectively. The prevalence of carbapenem-resistant Escherichia coli(CREC)and carbapenem-resistant Klebsiella pneumoniae(CRKP)were 1.3%(58/4 510)and 13.1%(307/2 340);62.1%(36/58)and 9.8%(30/307)of CREC and CRKP were resistant to ceftazidime/avibactam combination,respectively. The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)complex was 59.5%(241/405),while less than 5% of Acinetobacter baumannii complex was resistant to tigecycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa(CRPA)was 18.4%(98/534). There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of main Gram-negative bacteria resistance among different regions,with statistically significant differences in the prevalence of CRKP and CRPA( χ2=20.489 and 20.252, P<0.001). The prevalence of CREC,CRKP,CRPA,CRAB,ESBLs-producing Escherichia coli and Klebsiella pneumoniae were higher in provinicial hospitals than those in municipal hospitals( χ2=11.953,81.183,10.404,5.915,12.415 and 6.459, P<0.01 or <0.05),while the prevalence of CRPA was higher in economically developed regions(per capita GDP ≥ 92 059 Yuan)than that in economically less-developed regions(per capita GDP <92 059 Yuan)( χ2=6.240, P=0.012). Conclusions:The proportion of Gram-negative bacteria in bloodstream infections shows an increasing trend,and Escherichia coli is ranked in the top,while the trend of CRKP decreases continuously with time. Decreasing trends are noted in ESBLs-producing Escherichia coli and Klebsiella pneumoniae. Low prevalence of carbapenem resistance in Escherichia coli and high prevalence in CRAB complex have been observed. The composition ratio and antibacterial spectrum of bloodstream infections in different regions of China are slightly different,and the proportion of main drug resistant bacteria in provincial hospitals is higher than those in municipal hospitals.
4.Screening of characteristic genes in early-onset pre-eclampsia and analysis of their association with immune cell infiltration based on bioinformatics analysis and machine-learning algorithms
Zitong WU ; Yuanyuan ZHENG ; Xin DING
Chinese Journal of Perinatal Medicine 2024;27(1):51-61
Objective:To screen the characteristic genes of early-onset pre-eclampsia (EOSP) and to analyze their association with immune cell infiltration based on bioinformatics analysis and machine learning methods.Methods:In the Gene Expression Omnibus (GEO) database, the mRNA sequences of placental tissues from women with EOSP and normal pregnancy were retrieved using the term "early-onset pre-eclampsia". The R language was used for background correction, standardization, summarization, and probe quality control. Annotation packages were downloaded for ID conversion and the expression matrices were extracted. The differentially expressed genes (DEGs) between the EOSP and the normal pregnancy in the metadata were analyzed after correcting for batch effects using the limma package. Characteristic genes were identified through the support vector machine (SVM) -recursive feature elimination (RFE) method and the LASSO regression model. The area under the curve (AUC) was calculated to judge the diagnostic efficiency of the characteristic genes. Placental tissues were retrospectively collected for verification from 15 patients with EOSP and 15 with normal pregnancy who were delivered at Beijing Obstetrics and Gynecology Hospital, Capital Medical University from January 1, 2022, to February 28, 2023. The expression of characteristic genes was verified using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, which were further validated in the validation dataset. Finally, the CIBERSORT algorithm was used to analyze the relative proportion of infiltrating immune cell in EOSP. A t-test was used for differential analysis. Results:Three gene datasets were downloaded, including GSE44711 (eight cases each for EOSP and normal pregnancy), GSE74341 (seven cases for EOSP and five cases for normal pregnancy), and GSE190639 (13 cases each for EOSP and normal pregnancy). A total of 29 DEGs were screened after combining the GSE44711 and GSE74341 datasets, including 27 upregulated and two downregulated genes. Gene ontology enrichment analysis showed that these genes are mainly involved in the secretion of gonadotropins, female pregnancy, regulation of endocrine processes, secretion of endocrine hormones, and negative regulation of hormone secretion. Eight characteristic genes ( EBI3, HTRA4, TREML2, TREM1, NTRK2, ANKRD37, CST6, and ARMS2) were screened using the LASSO regression algorithm combined with SVM-RFE algorithm and the expression differences of these characteristic genes were verified as statistically significant by qRT-PCR and Western blot (all P<0.05, except for CST6). Logistic regression algorithm showed that the AUC (95% CI) of TREML2, ANKRD37, NTRK2, TREM1, HTRA4, EBI3, and ARMS2 were 0.979 (0.918-1.000), 0.969 (0.897-1.000), 0.969 (0.892-1.000), 0.979 (0.918-1.000), 0.990 (0.954-1.000), 0.990 (0.954-1.000), and 0.903 (0.764-1.000). Immune cell infiltration analysis indicated that the infiltration ratio of M2 macrophages in the placental tissue from EOSP was significantly lower than that in the normal pregnancy (0.167±0.074 vs. 0.462±0.091, P=0.002), but the infiltration ratios of monocytes and eosinophils were significantly higher (0.201±0.004 vs. 0.085±0.006; 0.031±0.001 vs. 0.001±0.000, both P<0.05). The correlation analysis between characteristic genes and infiltrating immune cells found that the seven characteristic genes were closely related to the immune cells (all P<0.05). Conclusion:Seven characteristic genes that are critical for the prediction and early diagnosis of EOSP are screened using bioinformatics analysis and machine-learning algorithms in this study, which provides new research targets and a basis for the prevention and treatment of preeclampsia in the future.
5.Item Screening of the Clinical Aided Decision Scheme for Stroke Simultaneous Treatment of Disease,Pulse and Syndrome Based on Delphi Method
Mengmeng DING ; Qiaosheng REN ; Jian CHEN ; Dahe QI ; Kexin WANG ; Yuanyuan CHEN ; Lingbo KONG ; Jingling CHANG
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(2):149-153
Objective To screen items of the Clinical Aided Decision Scheme for Stroke Simultaneous Treatment of Disease,Pulse and Syndrome;To provide reference for the formulation and improvement of the scheme.Methods The Delphi method was used to distribute two rounds of questionnaires to 60 experts in cerebropathy or neurology across the country.Statistical analysis was performed on the questionnaire results of the scheme's items,including the disease names,etiology and pathogenesis,syndrome characteristics,rules and regulations,representative prescriptions,acupuncture and other therapies,and preventive care.Results Totally 42 and 50 valid questionnaires were collected.The experts reached the consensus for the importance of etiology and pathogenesis,rules and regulations,acupuncture and other therapies,and preventive care.In the section on syndrome characteristics,items with low relevance or causing ambiguity were removed.Items that were no longer used in modern times and different prescriptions with the same name were removed from the representative prescriptions.The names of syndromes,rules and regulations were unified.Conclusion The experts generally reached the consensus for the importance of the Clinical Aided Decision Scheme for Stroke Simultaneous Treatment of Disease,Pulse and Syndrome.However,there are still some limitations that require further study and discussion.
6.Research Progress in the Clinical Application and Mechanisms of Acupuncture in the Treatment of Postoperative Acute Pain and Postoperative Complications
Yuanyuan DING ; Feng XU ; Xiangdong CHEN
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2024;53(5):681-686
Postoperative complications are the third leading cause of death worldwide,and failure to intervene in time can lead to serious consequences.Despite the continuous improvement in medical level in recent years,the incidences of postoperative acute pain,postoperative nausea and vomiting,postoperative urinary retention,and perioperative neurocognitive disorders are still high.Numerous studies have shown that acupuncture during the perioperative period can not only reduce preoperative anxi-ety and relieve postoperative pain but also reduce the incidence of postoperative nausea and vomiting.Acupuncture,a safe and ef-fective clinical therapy,is associated with few adverse reactions;thus,it has attracted increased attention from physicians and has been fully used in various clinical settings.However,at present,the specific mechanism by which acupuncture exerts the above effects is not clear,which limits its use to a certain extent.This article reviews the research progress of acupuncture in the treat-ment of postoperative acute pain and the prevention of various postoperative complications,as well as related mechanisms.
7.Bone densities and biomechanical properties on different layers of the trabecular bone in the proximal tibia
Yuanyuan QUAN ; Haicheng WANG ; Yanlin LI ; Kai DING ; Yifan ZHANG ; Jianzhi ZHANG ; Wei CHEN
Chinese Journal of Orthopaedic Trauma 2024;26(8):711-718
Objective:To investigate the correlations between the bone mineral densities and biomechanical properties on different layers of the cancellous bone in the proximal tibia.Methods:Quantitative CT was conducted of the 15 specimens of adult male tibia. Based on the artificial destruction levels at the trabecular bone on the tibial plateau, the 15 specimens were randomly divided into 3 groups ( n=5): group A (cancellous bone on the 1.5 cm layer below the articular cartilage), group B (cancellous bone on the 3.0 cm layer under the articular cartilage) and group C (cancellous bone on the 0 to 3 cm range of the subchondral bone). After standing positions were simulated in the 3 sets of specimens, they were connected to a biomechanical testing machine. Twenty-four sites were selected and subjected to a vertical load of 600 N. Strain values and overall displacement values of the specimens were recorded before and after trabecular bone destruction. The correlations were analyzed between bone density and displacement values in groups A and B. The strain values before and after trabecular bone destruction, as well as the overall deformation values of the specimens were compared between the 3 groups. Results:The bone densities of specimens in groups A and B were negatively correlated with the displacement values before and after destruction ( P<0.05). Comparisons of strain values at the 24 sites before and after trabecular bone destruction within 3 groups: There were statistically significant differences in the strain values at 8 sites between before and after trabecular bone destruction in group A ( P<0.05). Of the 8 sites, 6 showed increased strains which were mainly concentrated around the insertion point of the anterior cruciate ligament and the medial tibial plateau. There were statistically significant differences in the strain values between before and after trabecular bone destruction at 3 sites in group B ( P<0.05). The strains at all the 3 sites increased, mainly concentrated behind the surface below the level of destruction. There were statistically significant differences in the strain values at 10 sites in group C between before and after trabecular bone destruction ( P<0.05). Of the 10 sites, 5 showed a decrease in the strain which was concentrated above the destruction plane, and 5 showed an increase in the strain which was concentrated below the destruction plane. The overall deformation values of the specimens in groups A, B, and C were (0.033±0.003) mm, (0.015±0.003) mm, and (0.066±0.007) mm, respectively, showing statistically significant differences between the 3 groups ( P<0.05) as well as between any 2 groups ( P<0.05). Conclusions:Bone mineral density in the cancellous bone of the proximal tibia has some value in assessment of the bone strength. Destruction of the proximal tibial cancellous bone can significantly change the strain distribution on the proximal tibia. The proximal cancellous bone of the tibia plays a key role in stress support and load conduction.
8.National bloodstream infection bacterial resistance surveillance report(2022): Gram-positive bacteria
Chaoqun YING ; Yunbo CHEN ; Jinru JI ; Zhiying LIU ; Qing YANG ; Haishen KONG ; Haifeng MAO ; Hui DING ; Pengpeng TIAN ; Jiangqin SONG ; Yongyun LIU ; Jiliang WANG ; Yan JIN ; Yuanyuan DAI ; Yizheng ZHOU ; Yan GENG ; Fenghong CHEN ; Lu WANG ; Yanyan LI ; Dan LIU ; Peng ZHANG ; Junmin CAO ; Xiaoyan LI ; Dijing SONG ; Xinhua QIANG ; Yanhong LI ; Qiuying ZHANG ; Guolin LIAO ; Ying HUANG ; Baohua ZHANG ; Liang GUO ; Aiyun LI ; Haiquan KANG ; Donghong HUANG ; Sijin MAN ; Zhuo LI ; Youdong YIN ; Kunpeng LIANG ; Haixin DONG ; Donghua LIU ; Hongyun XU ; Yinqiao DONG ; Rong XU ; Lin ZHENG ; Shuyan HU ; Jian LI ; Qiang LIU ; Liang LUAN ; Jilu SHEN ; Lixia ZHANG ; Bo QUAN ; Xiaoping YAN ; Xiaoyan QI ; Dengyan QIAO ; Weiping LIU ; Xiusan XIA ; Ling MENG ; Jinhua LIANG ; Ping SHEN ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2024;17(2):99-112
Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-positive bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-positive bacteria from blood cultures in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System(BRICS)were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute(CLSI). WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:A total of 3 163 strains of Gram-positive pathogens were collected from 51 member units,and the top five bacteria were Staphylococcus aureus( n=1 147,36.3%),coagulase-negative Staphylococci( n=928,29.3%), Enterococcus faecalis( n=369,11.7%), Enterococcus faecium( n=296,9.4%)and alpha-hemolyticus Streptococci( n=192,6.1%). The detection rates of methicillin-resistant Staphylococcus aureus(MRSA)and methicillin-resistant coagulase-negative Staphylococci(MRCNS)were 26.4%(303/1 147)and 66.7%(619/928),respectively. No glycopeptide and daptomycin-resistant Staphylococci were detected. The sensitivity rates of Staphylococcus aureus to cefpirome,rifampin,compound sulfamethoxazole,linezolid,minocycline and tigecycline were all >95.0%. Enterococcus faecium was more prevalent than Enterococcus faecalis. The resistance rates of Enterococcus faecium to vancomycin and teicoplanin were both 0.5%(2/369),and no vancomycin-resistant Enterococcus faecium was detected. The detection rate of MRSA in southern China was significantly lower than that in other regions( χ2=14.578, P=0.002),while the detection rate of MRCNS in northern China was significantly higher than that in other regions( χ2=15.195, P=0.002). The detection rates of MRSA and MRCNS in provincial hospitals were higher than those in municipal hospitals( χ2=13.519 and 12.136, P<0.001). The detection rates of MRSA and MRCNS in economically more advanced regions(per capita GDP≥92 059 Yuan in 2022)were higher than those in economically less advanced regions(per capita GDP<92 059 Yuan)( χ2=9.969 and 7.606, P=0.002和0.006). Conclusions:Among the Gram-positive pathogens causing bloodstream infections in China, Staphylococci is the most common while the MRSA incidence decreases continuously with time;the detection rate of Enterococcus faecium exceeds that of Enterococcus faecalis. The overall prevalence of vancomycin-resistant Enterococci is still at a low level. The composition ratio of Gram-positive pathogens and resistant profiles varies slightly across regions of China,with the prevalence of MRSA and MRCNS being more pronounced in provincial hospitals and areas with a per capita GDP≥92 059 yuan.
9.Impacts of Artesunate on the Formation of Kidney Oxalate Stones by Regulating the PI3K/Akt Signaling Pathway
Yuanyuan WANG ; Ding LI ; Chuangui LI
Journal of Medical Research 2024;53(10):149-156,173
Objective To investigate the impacts of artesunate(Art)on the formation of kidney stones and the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)signaling pathway.Methods Rats were randomly divided into the Control group,Model group,Artesunate low-dose group(ART-L group),artesunate high-dose group(ART-H group),artesunate high-dose+PI3K in-hibitor group(Art+LY294002 group).The rat model of kidney stone was established by drinking water with ethylene glycol and injecting sodium oxalate intraperitoneally.A kidney stone cell model was established by exposing HK-2 cells to 200 mg/L calcium oxalate mono-hydrate crystals and 2 mmol/L oxalate.The levels of Ca2+and oxalic acid(Ox)in urine and serum creatinine(Scr)and blood urea ni-trogen(BUN)were detected;Pizzolato staining was applied to observe the deposition of calcium salts in renal tissues;the pathological morphology in renal tissues was detected by HE staining;malondialdehyde(MDA)and superoxide dismutase(SOD)kits were used to detect the levels of MDA and SOD in renal tissue and HK-2 cells;ELISA method was applied to detect TNF-α,IL-1βand IL-6 in renal tissue and HK-2 cells;fluorescence quantification was applied to detect the levels of Bax,caspase-3,PI3K and Akt;immunohis-tochemical and Western blot were applied to detect the expression levels of PI3K and Akt proteins in the renal tissues.HK-2 cell apopto-sis was measured by flow cytometry.Results Compared with the Model group,Ca2+,Ox,Scr,BUN,the level of MDA,TNF-α,IL-1 β,IL-6 in renal tissue and HK-2 cells,the expression of Bax,caspase-3 in renal tissue decreased in Art-L and Art-H groups,the level of SOD,PI3K,and Akt in renal tissue and HK-2 cells increased(P<0.05);compared with the Art-H group,Ca2+,Ox,Scr,BUN,the level of MDA,TNF-α,IL-1 β,IL-6 in renal tissue and HK-2 cells,the expression of Bax,caspase-3 in renal tis-sue increased in the Art+LY294002 group,the level of SOD,PI3K and Akt in renal tissue and HK-2 cells decreased(P<0.05).Conclusion Artesunate can inhibit the formation of kidney stones;its inhibition of stone formation may be produced through the mecha-nism of up-regulating the PI3K/Akt signaling pathway.
10.circDDX17 targets miR-223-3p / RIP3 to regulate the proliferation and apoptosis of non-small cell lung cancer cells
Chengzhi DING ; Guolei WANG ; Gongqian JIANG ; Hongtao WANG ; Yuanyuan LIU ; Haoliang ZHANG ; Fang SUN ; Li WEI
Chinese Journal of Oncology 2024;46(3):239-248
Objective:To explore the molecular mechanism of circDDX17 regulating the proliferation and apoptosis of non-small cell lung cancer cells by targeting the miR-223-3p/RIP3 molecular axis.Methods:The expression levels of circDDX17, miR-223-3p, and RIP3 in human normal lung epithelial cell lines BEAS-2B and non-small cell lung cancer cells H1299, A549, and H446 were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). The plasmids of pcDNA, pcDNA-circDDX17, anti-miR-con, anti-miR-223-3p, pcDNA-circDDX17 and miR-con, pcDNA-circDDX17 and miR-223-3p mimics were transfected into H1299 cells. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay was used to detect the cell proliferation. Flow cytometry was used to detect the cell cycle and cell apoptosis. Plate cloning experiment was used to detect cell proliferation ability. The dual luciferase report experiment was applied to verify the targeting relationship between miR-223-3p with circDDX17 and RIP3. Western blot was used to detect the protein expression of cyclinD1, CDK2, cleaved caspase-3 and Bax.Results:The expression levels of circDDX17 and RIP3 mRNA in H1299, A549, and H446 cells were significantly reduced ( P<0.05), the expression level of miR-223-3p mRNA was significantly increased ( P<0.05) compared with BEAS-2B. The cell viability [(69.46±4.68)%], the number of cell clones (83.49±7.86), the proportion of cells in S phase [(22.52±1.41) %], the protein expression levels of cyclinD1 and CDK2 in PCDNa-CircDDX17 group were lower than those in pcDNA group [(97.54±7.72)%, 205.03±13.37, (28.69±1.49)%, respectively, P<0.05], while the percentage of G 0/G 1 phase cells [(64.45±3.56)%], apoptosis rate [(18.36±1.63)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17 group were higher than those of pcDNA group [(51.33±2.76) % and (5.21±0.54) %, respectively, P<0.05]. The viability [(72.64±5.44)%], the number of cell clones (78.16±8.23), the proportion of S-stage cells [(21.34±1.59) %], the protein expression levels of CyclinD1 and CDK2 in anti-miR-223-3p group were lower than those in anti-miR-con group [(103.47±6.25)%, 169.32±14.53, (28.43±1.26)%, respectively, P<0.05]. Percentage of G 0/G 1 phase cells [(62.86±3.28)%], apoptosis rate [(14.64±1.67)%], the protein expression levels of cleaved caspase-3 and Bax in the anti-miR-223-3p group were higher than those of anti-miR-con group [(51.33±2.71)% and (4.83±0.39)%, respectively, P<0.05]. MiR-223-3p has complementary sites with circDDX17 or RIP3. The viability [(135.45±9.28)%], the number of cell clones (174.64±10.68), the proportion of S-phase cells [(26.39±2.25)%], the protein expression levels of cyclinD1 and CDK2 in pcDNA-circDDX17+miR-223-3p group were higher than those in pcDNA-circDDX17+miR-con group [(101.56±6.68)%, 107.65±7.62, (21.64±1.72)%, P<0.05]. Percentage of G 0/G 1 phase cells [(56.64±2.76)%], apoptosis rate [(8.34±0.76)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17+miR-223-3p group were lower than those of pcDNA-circDDX17+miR-con group [(64.03±3.48)% and (15.21±1.18)%, respectively, P<0.05]. Conclusion:circDDX17 could inhibit the proliferation and induce apoptosis of non-small cell lung cancer cells via targeting the miR-223-3p / RIP3 molecular axis.

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