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Table of Content

    05 January 2023, Volume 43 Issue 1
    Invited Reviews: Basic Research and Clinical Translation of Adult Stem Cells
    Immunomodulatory plasticity of mesenchymal stem cells and the strategy of clinical application
    WANG Haiyan, HAN Qin, ZHAO Chunhua
    2023, 43(1):  2-11.  doi:10.16352/j.issn.1001-6325.2023.01.0002
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    Mesenchymal stem cells (MSCs) are pluripotent stem cells with self-renewal, multi-directional differentiation and immunoregulatory capabilities. More and more studies have demonstrated the beneficial role of MSCs in the treatment of immune disorders. However, there is heterogeneity in the immunoregulatory capacity of MSCs obtained in vivo or cultured in vitro, and the immunoregulatory capacity of MSCs is also regulated by the microenvironment. This review provides an overview of the mechanisms of the immunosuppressive and immune supportive abilities of MSCs, introduces recent studies on the immunoregulatory mechanisms of MSCs and the plasticity hypothesis of their immunoregulatory abilities, and discusses strategies to enhance the immunoregulatory abilities of MSCs in the cell production stage. This review aims to provide a reference for the clinical translational research of MSCs.
    Research progress of mesenchymal stem cells in the treatment of liver disease
    ZHOU Jiahang, CHEN Junyao, CAO Hongcui
    2023, 43(1):  12-20.  doi:10.16352/j.issn.1001-6325.2023.01.0012
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    Liver disease is a serious threat to human health. However, there is still no effective medical treatment for end-stage liver diseases such as cirrhosis and liver failure. Mesenchymal stem cells (MSCs) are expected to be used clinically for liver cirrhosis caused by various etiologies, acute-on-chronic liver failure, chronic liver failure, and complications post-liver transplantation by their multi-directional differentiation and immunomodulatory abilities. MSCs can directly differentiate into hepatocyte-like cells and indirectly secrete soluble factors to achieve goals such as antioxidant, anti-regulated cell death, anti-fibrosis, inhibiting inflammation, inhibiting immune response, etc., so to reduce liver damage and improve survival rate. A sound quality standard evaluation system and a more in-depth mechanism exploration are the prerequisites for MSCs to be used in the clinical treatment of liver diseases in the future.
    Progress in application of mesenchymal stem cells in treatment of acute myocardial infarction
    JIANG Yu, QIAN Haiyan
    2023, 43(1):  21-29.  doi:10.16352/j.issn.1001-6325.2023.01.0021
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    Acute myocardial infarction (AMI) is an ischemic myocardial necrosis caused by coronary artery occlusion, which can't be self-repaired through cardiomyocyte regeneration. The necrotic myocardium is replaced by fibrous scars during ventricular remodeling, ultimately leading to heart failure. The safety and efficacy of mesenchymal stem cells (MSCs) transplantation after AMI have been demonstrated by numerous preclinical and clinical studies. MSCs are composed of heterogeneous cells with multi-directional differentiation potential, which can regulate oxidative stress, and secrete a variety of cytokines and growth factors. After being implanted in vivo, MSCs play their roles in immunomodulatory, angiogenesis, anti-inflammatory, and anti-apoptosis through trans-differentiation, cell fusion, and paracrine. Currently, there has been a variety of access to implant MSCs, including intramyocardial injection, intracoronary infusion, and intravenous injection. In addition, the dose and timing of transplantation are also important factors affecting the therapeutic effect of MSCs. However, the low retention and survival rates of MSCs in infarcted myocardium after being transplanted limit their further effect and hinder their clinical translation. In recent years, new concepts, strategies, technologies, and methods for MSCs treatment have been proposed, including cell preconditioning, optimization of the infract local microenvironment, combined gene therapy or tissue engineering technology, exosome infusion, and targeted transplantation of stem cells and their exosomes, which significantly improve the transplantation efficiency and therapeutic effect of MSCs, and open a new chapter for the research and transformation of stem cells to repair the infarcted myocardium. This article reviews the progress of MSCs in repairing myocardial infarction in recent years.
    Progress in application of mesenchymal stem cells in treatment of nephropathy
    YANG Yunzhao, SHI Meihan, ZHOU Cheng, BAI Xueyuan
    2023, 43(1):  30-37.  doi:10.16352/j.issn.1001-6325.2023.01.0030
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    Mesenchymal stem cells (MSCs) can reduce inflammation, restore renal injury and maintain immune homeostasis by paracrine various cytokines and regulating immune cells. In preclinical studies, it has been proved that mesenchymal stem cells have obvious therapeutic effects on acute kidney injury, diabetic nephropathy, lupus nephritis and various glomerular diseases in animal models. In clinical studies, mesenchymal stem cells have also been shown to be safe and partially effective. Mesenchymal stem cell therapy has revealed a new model of biotherapy in kidney diseases.
    Research advances in application of mesenchymal stem cells for aging related diseases
    SHANG Lipeng, LIU Guoxiang, DING Xiaoyan, LI Bing, ZHAO Chunhua, LI Xiaoxia
    2023, 43(1):  38-45.  doi:10.16352/j.issn.1001-6325.2023.01.0038
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    Aging related diseases seriously affect the health of the elderly. Studies on the roles and mechanisms of mesenchymal stem cells (MSCs) in aging related diseases have attracted more and more attention. Besides, a number of relative clinical trials have been carried out in the domestic and overseas. This review summarizes the roles, molecular mechanisms, clinical trials of MSCs and MSCs-derived exosomes in several typical age-related diseases such as osteoporosis, Alzheimer's disease, Parkinson's disease, type 2 diabetes, atherosclerosis, chronic obstruc-tive pulmonary disease and tumor. This review may provide some theoretical reference and advices for future studies of MSCs in aging related diseases.
    Interpretation of the essence of mesenchymal stem cells with the “kidney-triple energizers” system
    ZHANG Xiaotong, ZHAO Yuechun, ZHU Rongjia, ZHAO Chunhua, SONG Ping
    2023, 43(1):  46-50.  doi:10.16352/j.issn.1001-6325.2023.01.0046
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    Kidney and triple energizers are inseparable in structure and function. Therefore, it is proposed that “kidney-triple energizers” might be the essence of mesenchymal stem cells (MSCs) system. As a cardinal, this article aims to interpret the theoretical connotation of the mesenchymal stem cell system from the perspective of “the kidney essence is the body and the triple energizers is the function”. It is reflected in the kidney-three energizers system, and guided by the theory of traditional Chinese medicine, the modern technology of MSCs has achieved good results in the application of refractory diseases such as COVID-19, and a preliminary exploration of the integration of Chinese and Western medicine research has been carried out.
    Original Articles
    Expression and enzymatic activity analysis of non-structure protein 14 of SARS-CoV-2
    CHEN Shuang, XU Jingwen, DING Kaiyun, LIU Jiansheng, PENG Xiaozhong, LIU Hongqi
    2023, 43(1):  51-58.  doi:10.16352/j.issn.1001-6325.2023.01.0051
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    Objective To obtain the non-structure protein 14 (Nsp14) of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) with higher purity and enzymatic activity. Methods This study firstly analyzed the rare codons in the gene of nsp14 according to the codon usage bias of E. coli, followed by codon optimization. The optimized nucleotide fragment of nsp14 was cloned into four kinds of expressing vectors respectively. Comparative analysis of yield and solubility was performed among these expressed four fusion proteins. The best one was chosen for further optimization of expressing conditions. After the fusion protein was purified by glutathione affinity column, the fusion tag was removed by 3C protease, and then the protein was purified by glutathione affinity column and molecular sieve column for further analysis of enzymatic activity through urea polyacrylamide gel electrophoresis. Results Many rare codons were found in expression of SARS-CoV-2 nsp14 in E. coli, among which some rare codons were distributed in close range and tandem. The best recombinant plasmid for expressing the fusion protein was pGEX6P1-GST-OPTI-Nsp14, which gave an eptimal expression in 30 ℃ with high yield and solubility. After purification, a higher purity of Nsp14 with nuclease activity was obtained. Conclusions This study shows that the SARS-CoV-2 Nsp14 protein with nuclease activity is successfully prepared, which lays a foundation for the further research on the structure and function of SARS-CoV-2 Nsp14, and provides favorable conditions for screening antiviral drugs targeting at Nsp14 of SARS-CoV-2.
    Production of monoclonal antibodies specific for the endogenous retrovirus Gag antigen expressed in pancreatic islets of mice
    LI Yu, ZHANG Wen, YUAN Yu, LI Shuting, WANG Zhe,BEN Yali, DAI Yang
    2023, 43(1):  59-67.  doi:10.16352/j.issn.1001-6325.2023.01.0059
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    Objective To explore a newly discovered T cell antigen [the group-specific antigen(Gag)of endogenous retrovirus (ERV)] associated with type 1 diabetes (T1D) and the pathogenesis of T1D. Methods First, to analyze the amino acid sequence of an islet-expressing Gag antigen, Gag 194 and identified one peptide[P193-P210(VSRLRGRRDPPAVDSTTS)], for production of Gag-specific monoclonal antibodies (mAbs). Second, ELISA and Western blot methods were performed to evaluate antigen specificity of the mAbs. Third, the expression of Gag antigen in pancreatic islets of T1D-susceptible non-obese diabetic (NOD) mice and T1D-resistant C57BL/6 mice was examined by immunohistochemical experiments. Results Five mAbs, namely 1F4C8,2D4C6,3C3B9,4D6B3 and 5F9E4 were obtained, and all of them were IgG2b subtype. Their antigen specificity and titer varied slightly. Three mAbs (1F4C8, 2D4C6 and 5F9E4) specifically recognize the ERV Gag antigen expressed in the pancreatic islets of NOD mice and at the age of 3 weeks, the antigen was detectable. Within the islets, the Gag antigen located near β-cells rather than the area infiltrated by lympho- cytes.In contrast, the T1D-resistant C57BL/6 mice do not express the Gag antigen as detected using the same mAb. Conclusions The expression of ERV Gag antigen in pancreatic islets of NOD mice occurs at very young age, suggesting that genetic factors of this diabetes-prone mouse strain may contribute to the expression.
    Relationships between progrosis, immune infiltration and expression of SMYD3 in pan-cancer
    CHEN Haoran, SI Tian, JIANG Yishen, WANG Dianheng, ZHOU Yabo, LYU Jiadi
    2023, 43(1):  68-75.  doi:10.16352/j.issn.1001-6325.2023.01.0068
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    Objective The study focuses on the prognostic value of SMYD3 in pan cancer, and analyze the influence of SMYD3 on tumor tissue immune infiltration. Methods The gene expression and protein density data were collected from TCGA, GTEx and HPA databases. Kaplan-Meier(KM) algorithm was used to evaluate the impact of SMYD3 on the prognosis of patients with different kinds of tumor. The ESTIMATE algorithm and Timer database were applied to investigate the relationship between SMYD3 and immune microenvironment. And the results were verified in B16 cell line by knockout SMYD3. Results Higher expression of SMYD3 was observed in tumor tissue than normal tissue in both mRNA and protein level. And SMYD3 was thought to be associated with worse outcomes of patients(P<0.05) and less immune cells inflation in tumor tissue(P<0.05). Knockout Smyd3 in B16 cell line increased the expression of antigen present genes(P<0.05). Conclusions Smyd3 is an important gene related to the prognosis of patients and the antigen presentation function of tumor cells. Smyd3 can inhibit the infiltration of immune cell by down-regulation of antigen-present genes, which induces the immune escape.
    Imaging of breast cancer targeted by the nucleic acid aptamer W3-quantum dots
    WANG Qun, LI Xin, MA Jun, LI Wanming
    2023, 43(1):  76-82.  doi:10.16352/j.issn.1001-6325.2023.01.0076
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    Objective To perform quantum dots(QDs)imaging of breast cancer cells and breast cancer tissue paraffin specimens by using nucleic acid aptamer W3 binding specificity on breast cancer cells. Methods The binding specificity of nucleic acid aptamer W3 to breast cancer cell lines was detected by flow cytometry, and based on the binding reaction principle of biotin and streptomyces avidin, the quantum dot targeting imaging of nucleic acid aptamer W3 on breast cancer cell lines and tissue specimens of clinical patients was observed by fluorescence microscopy, and its clinical significance was further analyzed. Results The nucleic acid aptamer W3 specifically binded to breast cancer cell lines with high malignancy,and after coupling quantum dots QD605(W3-QDs), it achieved targeted imaging of breast cancer cell lines and breast cancer tissues with high malignancy, and the fluorescence intensity of breast cancer tissues with high malignancy degree was significantly higher than that of breast cancer tissues with low malignancy. Conclusions The nucleic acid aptamer W3 may achieve target imaging of breast cancer after coupling quantum dots, which can be used for early diagnosis of breast cancer.
    Effect of osteopontin in pathogenesis of hepatic ischemia-reperfusion injury of rats
    GUO Liping, SU Juan, TIAN Xinyan, LU Li, HU Yarong, QIN Yan
    2023, 43(1):  83-86.  doi:10.16352/j.issn.1001-6325.2023.01.0083
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    Objective To study the role of osteopontin (OPN) during hepatic ischemia-reperfusion injury (HI/RI) in rats. Methods The rats were divided into sham operation group (sham) and ischemia-reperfusion (I/R) 6, 12 and 24 h groups, with 6 rats in each group. HE staining was used to observe the morphological changes of liver tissue. The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in plasma, the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in liver tissue were measured by colorimetric method. The expression of OPN mRNA in liver tissue was determined by RT-qPCR. The content of OPN in liver tissue was determined by ELISA. Results Compared with the sham group, HE staining showed obvious degeneration and necrosis of hepatocytes after I/R, which was most obvious in the I/R 24 h group. The plasma levels of AST and ALT increased, most significantly at I/R 12h(P<0.01). The content of MDA in liver tissue increased, and the activity of SOD decreased, which was most significant at I/R 24 h(P<0.01). The expression of OPN mRNA in liver tissue was significantly increased in I/R groups(P<0.01). OPN content was also significantly increased in I/R 24 h group(P<0.05). Conclusions OPN is involved in the pathogenesis of HI/RI in rats and is a potential indicator applied in the evaluation of liver injury.
    miR-203a-3p inhibits proliferation of human hepatocellular carcinoma cell lines
    HUANG Yisheng, YE Qiwen, WANG Jingfeng
    2023, 43(1):  87-94.  doi:10.16352/j.issn.1001-6325.2023.01.0087
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    Objective To explore the molecular mechanism of regulation on the proliferation and apoptosis in hepatocellular carcinoma(HCC) cells by miR-203a-3p. Methods RT-qPCR was used to detect the expression of miR-203 a-3p in the sample of liver cancer tissues and liver cancer cell lines. The HepG2 cell model expressing miR-203a-3p was established. Cell proliferation was observed by MTT method and the apoptosis of cells was detected by flow cytometry. Plate colony formation assay was used to detect clone formation and cell scratch assay was used to detect cell migration. Expression profile chip was applied in the detection of mRNA and ELISA was used to detect the secretion of transforming growth factor-β1 (TGF-β1). TGF-β1 protein expression was examined by Western blod technology. Results miR-203a-3p was weekly expressed in HCC tissues (P<0.001)and was negatively correlated with tumor stage (P<0.05). Over-expression of miR-203a-3p in HepG2 cells significantly reduced cell proliferation (P<0.05), migration (P<0.05), clone formation (P<0.05) and promoted cell apoptosis(P<0.05); TGF-β1 expression decreased(P<0.05) while the expression of TGFβR1 increased (P<0.05); The content of TGF-β1 in the cell culture supernatant decreased (P<0.05). The protein of TGF-β1 in cells decreased (P<0.05) and TGF-β1 recombinant protein reversed the miR-203a-3p induced inhibition of proliferation as well as migration (P<0.05). Conclusions miR-203a-3p inhibits proliferation of human hepatocellular carcinoma cell lines, promotes apoptosis and inhibits the expression of TGF-β1 protein.
    miR-504 regulates proteasome activator and inhibits the proliferation and migration of breast cancer cell line MCF-7
    YANG Liangquan, YU Miao, JIANG Qian, ZHANG Minghui, YANG Haisong
    2023, 43(1):  95-101.  doi:10.16352/j.issn.1001-6325.2023.01.0095
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    Objective To observe the regulation of miR-504 in mouse models and the biological behavior of breast cancer cells, find the key target genes of miR-504 in the regulation process, and explore the working mechanism of miR-504 in breast cancer. Methods A mouse model of breast cancer was established and the effect of miR-504 on tumor growth was observed. Target genes of miR-504 were screened by Target Scaning and confirmed by double luciferase reporter gene assay. RT-qPCR and Western blot were used to detect proteasome activator complex subunit 3(PSME3) expression in breast cancer and adjacent tissues. The effects of miR-504 and PSME3 on proliferation, cycle and migration of human breast cancer cell line MCF-7 were determined by CCK8 assay, flow cytometry and Transwell assay. Results It was found that miR-504 inhibited tumor growth in nude mouse xenograft model of breast cancer(P<0.01). PSME3 was a target gene of miR-504, and its expression in breast cancer tissues was significantly increased. Up-regulation of miR-504 expression could inhibit the expression of PSME3 (P<0.01). Compared with NC group, miR-504 mimic group could inhibit the proliferation, S-phase arrest and migration ability of breast cancer cells, while compared with miR-504 mimic group, miR-504 mimic+PSME3 group, increased proliferation the proportion of S-phase and migration ability(P<0.01)in MCF-7 cell. Conclusions miR-504 may play an anti-cancer role in breast cancer by targeting PSME3 regulation.
    LncRNA MINCR inhibits LPS-induced damage and inflammation of alveolar epithelial cell line A549 by targeting at miR-223
    YANG Guanglin, REN Lijuan, CHEN Wenjing, XIONG Weijun
    2023, 43(1):  102-109.  doi:10.16352/j.issn.1001-6325.2023.01.0102
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    Objective To investigate the potential effects of long non-coding RNA (lncRNA) MINCR on lipopolysaccharide (LPS)-induced human alveolar epithelial cell line A549 damage and inflammation through targeted regulation of miR-223. Methods A549 cells were randomly divided into control group, LPS group, LPS+transfection group(Si NC group, Si MINCR group, miR-223 NC group, miR-223 mimic group, Si NC+miR-223 NC group, Si NC+miR-223 antagomir group, Si MINCR+miR-223 NC group, and Si MINCR+miR-223 antagomir group). RT-qPCR was performed to determine the expression levels of MINCR and miR-223; Flow cytometry was performed to determine the rate of apoptosis; ELISA was performed to determine the levels of serum tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6); Bioinformatics prediction and dual luciferase reporter gene assay were performed to verify the targeting relationship of MINCR and miR-223; Western blot was performed to test the expression levels of activated caspase-3 (cleaved caspase-3) and B-cell lymphoma-2(Bcl-2) proteins in the cells. Results There was a targeting relationship between MINCR and miR-223. Compared with the control group, the apoptosis rate, the level of TNF-α and IL-6, and the expression of MINCR, cleaved caspase-3 proteins in the LPS group were significantly increased (P<0.05).The expression of miR-223 and Bcl-2 proteins significantly decreased(P<0.05). Compared with the Si NC group, the apoptosis rate, the levels of TNF-α and IL-6, and the expression of MINCR and cleaved caspase-3 proteins in the Si MINCR group were obviously reduced (P<0.05); And the expression of Bcl-2 protein was obviously increased (P<0.05). Compared with miR-223 NC, the apoptosis rate, the levels of TNF-α and IL-6, and the expression of cleaved caspase-3 protein in the miR-223 mimic group were obviously reduced (P<0.05); And the expression of miR-223 and Bcl-2 proteins was obviously increased (P<0.05). miR-223 antagomir was able to reverse the effect of Si MINCR (P<0.05). Conclusions Silencing the expression of lncRNA MINCR may target the activation of miR-223 expression, inhibit cell apoptosis and inflammation, and reduce LPS-induced damage of alveolar epithelial cells.
    Acinetobacter baumannii outer membrane vesicles induced inflammation of human monocyte-derived macrophages THP-1
    FU Shixiang, XI Yue, DING Longkun, YAN Man, ZHAO Jun, JIAO Yudong, WU Liang
    2023, 43(1):  110-115.  doi:10.16352/j.issn.1001-6325.2023.01.0110
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    Objective To establish an ultra-filtration centrifugation method to purify the outer membrane vesicles (OMVs) of Acinetobacter baumannii(A.baumannii), and to investigate the ability of OMVs to induce inflamma-tion in human monocyte-derired macrophages THP-1. Methods A.baumannii was cultured, and the OMVs were purified from the supernatant of culture medium by ultra-filtration centrifuge,the morphology was observed by transmission electron microscopy. THP-1 cells were incubated with OMVs at the concentration of 5 and 50 μg/mL respectively. The expression of NOD-like receptor protein 3(NLRP3) and caspase-1 were determined by RT-qPCR. The concentration of IL-1β in culture medium supernatant was measured by ELISA. The expression of LC-3Ⅱ and Beclin-1 was examined by Western blot. The activity of caspase-3 was determined by enzyme dynamics assay. The expression of reactive oxygen species (ROS) was determined by flow cytometry. Results A.baumannii OMVs was purified successfully. The OMVs could activate NLRP3 inflammasome, increase the expression of LC-3Ⅱ and Beclin-1 and up-regulate the activity of caspase-3 (P<0.05). The OMVs significantly up-regulated the expression of ROS in THP-1 cells after 1 h co-culture (P<0.05). Conclusions OMVs of A.baumannii can significantly activate macrophages NLRP3 inflammasome and induce cell autophagy and apoptosis.
    Metformin inhibits the apoptosis of intervertebral disc tissue cells in rabbit model with lumbar intervertebral disc degeneration
    DONG Junli, CAI Shaokang, YAN Lei, CAI Yi
    2023, 43(1):  116-122.  doi:10.16352/j.issn.1001-6325.2023.01.0116
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    Objective To investigate the impacts of metformin on autophagy and inflammation of intervertebral disc tissue in rabbits, so as to preliminarily explore the molecular mechanism of its anti-intervertebral disc degeneration. Methods The rabbits were diveded into: sham operation group, model group, metformin group, autophagy inhibitor group and metformin + autophagy inhibitor group, with 10 rabbits in each. The positive expression of autophagy-related protein 7 (Atg7) was measured by immunohistochemical staining; The expression of autophagy-related proteins LC3Ⅱ, Beclin1, lysosome-associated membrane protein 2 (LAMP2), tumor necrosis factor-α (TNF-α), IL-1β, and apoptosis-related protein caspase-8 was measured by Western blot. Results Compared with the model group, the intervertebral disc degeneration of the rabbits in the metformin group was alleviated; The formation of autophagosomes and autolysosomes increased. The expression of autophagy proteins increased; And the expression of inflammation and apoptosis proteins decreased (P<0.05). Autophagy inhibitors could inhibit autophagy, aggra- vate intervertebral disc degeneration, increased the expression of inflammatory and apoptotic proteins, and attenuate the autophagy-enhancing, inflammation and apoptosis-enhancing effects by metformin (P<0.05). Conclusions Metformin may inhibit the apoptosis of intervertebral disc tissue and improve intervertebral disc degeneration by promoting autophagy activation and attenuating inflammatory response.
    Scutellaria Extract inhibits IL-22-induced hyperproliferation of human keratinocyte cell line HaCaT
    ZENG Ying, FU Guili
    2023, 43(1):  123-129.  doi:10.16352/j.issn.1001-6325.2023.01.0123
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    Objective To explore the experimental study of Scutellaria extract on IL-22-induced proliferation, apoptosis and inflammatory response of human keratinocyte cells line (HaCaT) via miR-369-3p. Methods HaCaT cells were divided into control group, IL-22 group (cell models, 100 ng/mL of IL-22 cultured cells), low, medium and high dose group (Scutellaria extract 50, 100, 150 μg/mL), anti-miR-NC intervention group (transfected anti-miR-NC), anti-miR-369-3p intervention group (transfected with anti-miR-369-3p), miR-NC+high-dose group (transfected with miR-NC, Scutellaria extract cultured cells), miR-369-3p+high-dose group (transfected with miR-369-3p, Scutellaria extract cultured cells). MTT experiment detected cell proliferation; Flow cytometry detected cell apoptosis; Western blot detected cleaved caspase-3 protein expression;ELISA detected TNF-α and IL-6 content; RT-qPCR detected miR-369-3p expression. Results Compared with the control group, the IL-22 group cell survival rate, miR-369-3p expression, TNF-α, IL-6 expression increased(P<0.05), and the difference in the cell apoptosis rate, cleaved caspase-3 protein expression were not signficantly(P>0.05); Compared with the IL-22 group, the cell survival rate, miR-369-3p expression, TNF-α, IL-6 expression decreased in the low, middle and high dose groups, while the apoptosis rate, cleaved caspase-3 protein expression increased(P<0.05). Compared with the anti-miR-NC intervention group, the anti-miR-369-3p intervention group had lower cell viability, TNF-α, IL-6 expression, while apoptosis rate, cleaved caspase-3 protein expression increased(P<0.05). Over-expression of miR-369-3p could partially reverse the effects of Scutellaria extract on cell viability, apoptosis and inflammatory response of psoriasis cell models. Conclusions Scutellaria extract inhibits IL-22-induced hyperproliferation and inflammatory response of human keratinocytes, and promotes apoptosis with potential mechanisms by inhibiting the expression of miR-369-3p.
    FOXP1 expression is increased after Stau1 knockdown of murine preadipocyte cell line 3T3-L1
    MENG Xuanyu, LIU Dihui, JIANG Shuo, GUAN Yaqun, LIANG Xiaodi
    2023, 43(1):  130-136.  doi:10.16352/j.issn.1001-6325.2023.01.0130
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    Objective To screen the differentially expressed genes after knocking down double-stranded RNA binding protein stanfen 1(Stau1)during the differentiation process of murine preadipocyte cell line 3T3-L1, analyze their biological functions, and use qPCR to verify the sequencing results for analysis. Methods RNA-seq was used to analyze the transcriptional regulation of genes after Stau1 knockdown. The STAU1 shRNA was constructed and transfected into 3T3-L1 cells. The cocktail method induced them to differentiate into mature adipocytes. The cells of 0 and 4 days were collected to establish a control cell group and a STAU1 knockdown group (3 groups of biological replicate samples) for a total of 12 groups of samples. Cell gene chip data set, with change log2 (Fold change)> 1 and corrected P<0.05 as the screening criteria to screen the differentially expressed genes between Stau1 knockdown cells and control group samples, and then the fat after knockdown Gene ontology (GO) and metabolic pathway analysis (KEGG pathway database) were performed on the differentially expressed genes of cells and control groups. Luciferase experiments confirmed the presence of SBS binding to forkhead box P1(FOXP1) mRNA 3′UTR. Results Compared with the control, STAU1 knockdown cell group, a total of 588 differentially expressed genes were screened, of which 406 were up-regulated and 182 were down-regulated. Differentially expressed genes were mainly involved in lipid metabolism and inflammatory responded factors in the biological process, and the main enriched signal pathways were related to sugar metabolism and energy metabolism. FOXP1 expression increased 5.3 times after knocking down Stau1, and the software predicted that the 3′UTR region of FOXP1 mRNA contained STAU1 binding sites. Conclusions Therefore, it is speculated that STAU1 can bind to the STAU1 binding site loacated at FOXP1 mRNA and promote its degradation.
    Propofol attenuates hypoxia-induced inflammation and apoptosis in rat pheochromocytoma cell line PC12
    GU Wei, ZHU Jianping, WU Pinwen
    2023, 43(1):  137-143.  doi:10.16352/j.issn.1001-6325.2023.01.0137
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    Objective To investigate whether propofol inhibits the expression of miR-141-3p and reduces the molecular mechanism of hypoxia-induced inflammation and apoptosis of pheochromocytoma cell line PC12. Methods PC12 cells were divided into control group, hypoxia group, (5, 10, 20)μmol/L propofol+hypoxia group, anti-miR-con+hypoxia group, anti-miR-141-3p+hypoxia group, miR-con+20 μmol/L propofol+hypoxia group, miR-141-3p+20 μmol/L propofol+hypoxia group. Flow cytometry was used to detect the apoptosis of PC12 cells; Western blot was employed to determine the expression of activated cleaved caspase-3 (cleaved caspase-3) protein, and the kits were implemented to monitor malondialdehyde (MDA) content and superoxide dismutase (SOD) activity; Reactive oxygen species fluorescent probe DCFH-DA method to determine reactive oxygen species (ROS) content; ELISA kits to assay tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 content, RT-qPCR to detect the expression of miR-141-3p. Results Compared with the control group, the apoptosis rate, cleaved caspase-3 protein expression level, MDA, ROS, TNF-α, IL-1β, IL-6 content and miR-141-3p expression of PC12 cells in hypoxia group were all increased, while SOD activity weakened (P<0.05). Compared with the hypoxia group, the apoptosis rate, cleaved caspase-3 protein expression level, MDA content, ROS, TNF-α, IL-1β, IL-6 content and miR-141-3p expression of PC12 cells in 5, 10, 20 μmol/L propofol+hypoxia group decreased with the increase of propofol concentration, and SOD activity increased with the increase of propofol concentration (P<0.05). The expression of miR-141-3p, cleaved caspase-3 protein expression, MDA, ROS, TNF-α, IL-1β, IL-6 content and apoptosis rates of PC12 cells in the anti-miR-141-3p+ hypoxia group were lower than those in the anti-miR-con+hypoxia group, and the SOD activity was higher than that in the anti-miR-con+hypoxia group (P<0.05). The miR-141-3p expression, cleaved caspase-3 protein expression level, MDA content, ROS, TNF-α, IL-1β, IL-6 content and cell apoptosis rate of PC12 cells in miR-141-3p+20 μmol/L propofol+hypoxia group were higher than miR-con+20 μmol/L propofol+hypoxia group, while SOD activity was lower than those from miR-con+20 μmol/L propofol+hypoxia group (P<0.05). Conclusions Propofol can alleviate the inflammatory response, oxidative stress and apoptosis of PC12 cells induced by hypoxia by inhibiting the expression of miR-141-3p.
    LncRNA DANCR reduces anoxia-induced rat myocardial cell line H9c2 injury by targeting miR-3646
    QIU Libin, ZHANG Xincai, ZHAO Qian
    2023, 43(1):  144-151.  doi:10.16352/j.issn.1001-6325.2023.01.0144
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    Objective To study the role of long non-coding RNA (lncRNA) differentiation antagonizing non-coding RNA (DANCR) in myocardial ischemia and the targeted regulation of microRNA-3646 (miR-3646). Methods H9c2 cells(rat cardiomyocytes cell line)were separated into 6 groups: control group, anoxia group, pc-DANCR (DANCR over-expression) group, pc-NC (DANCR over-expression control) group, miR-3646 mimic (miR-3646 mimicry) group, mimic NC (miR-3646 mimic control) group, and pc-DANCR+miR-3646 mimic group; RT-qPCR method was implemented to measure the expression of lncRNA DANCR and miR-3646 to determine the transfection effect; Flow cytometry was performed to measure the rate of apoptosis; CCK-8 assay was implemented to cell measure survival; Electron microscopy was applied to observe the structural injury of H9c2 cells; Western blot was performed to measure the expression of activated cleaved caspase-3 (cleaved caspase-3) and heme oxygenase-1 (Ho-1). Dual-luciferase experiments were performed to verify the targeting relationship of miR-3646 with lncRNA DANCR and Ho-1. RNA interference Ho-1 (ShHo-1) and pc-DANCR were co-transfected into H9c2 cells, and the cell viability was measured. Results Compared with the control group, the injury and apoptosis of H9c2 cells in the anoxia group were serious, the survival rate and the expression of lncRNA DANCR were decreased, and the expression of miR-3646 was increased (P<0.05). Compared with the anoxia group, the injury and apoptosis of H9c2 cells in the pc-DANCR group were alleviated, the survival rate was increased, the expression of anti-injury protective factor Ho-1 was increased, and the expression of miR-3646 was decreased(P<0.05). The injury and apoptosis of H9c2 cells in the miR-3646 mimic group were further aggravated, and miR-3646 mimic was able to attenuate the anti-injury and anti-apoptotic effects of pc-DANCR (P<0.05). There was a targeted regulatory relationship between lncRNA DANCR, Ho-1 and miR-3646. Interfering with Ho-1 attenuated the promoting effect of lncRNA-DANCR on the survival of H9c2 cells induced by anoxia (P<0.05). Conclusions LncRNA DANCR can down-regulate the expression of miR-3646 and promote the expression of Ho-1 to alleviate anoxia-induced H9c2 cells injury and improve cell survival.
    Dexmedetomidine attenuates anoxia- induced injury of human neuroblastoma cell line SH-SY5Y
    YAN Chen, LIU Tao, XUAN Fei
    2023, 43(1):  152-158.  doi:10.16352/j.issn.1001-6325.2023.01.0152
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    Objective To study the protective effect of dexmedetomidine (DEX) on human neuroblastoma cell line SH-SY5Y under anoxia condition and its specific mechanism. Methods SH-SY5Y cells were divided into control group, anoxia group (treated with final concentration of 1.2 mmol/L CoCl2 for 24 h) and DEX intervention group (treated with final concentration of 1.2 mmol/L CoCl2 and 10 μmol/L DEX for 24 h). Cell morphology was observed by microscope; Cell viability and apoptosis were detected by MTT assay, TUNEL staining and annexin V-FITC/PI staining; The expression levels of cleaved caspase-9, cleaved caspase-3, p75NTR and p-NF-κB proteins were detected by Western blot. Results Compared with control group, the cell size in anoxia group was shrinked(P<0.01). Cell viability was decreased (P<0.01). The apoptosis rate was increased (P<0.01). The expression levels of cleaved caspase-9, cleaved caspase-3, p75NTR and p-NF-κB proteins was all significantly increased (P<0.01). DEX intervention could significantly alleviate the above changes in anoxia group (P<0.01). Conclusions Alleviation of anoxia-induced SH-SY5Y cell injury is potentially mediatd by inhibiting p75NTR expression and NF-κB activity.
    Salvianolic acid B alleviates cartilage tissue damage in rat models with osteoarthritis
    QIAN Ligang, SUN Bo, YU Bo, CHEN Kang, MA Zheng, CHEN Taoping, WANG Yunfei
    2023, 43(1):  159-164.  doi:10.16352/j.issn.1001-6325.2023.01.0159
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    Objective To explore whether salvianolic acid B affects cartilage tissue damage, hypoxia-inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) in osteoarthritis rats. Methods A rat model of osteoarthritis was constructed, and rats were randomly divided into sham operation group, osteoarthritis group(anterior cruciate ligment transection, ACLT), nimesulide group, salvianolic acid B high (40 mg/kg), and low (10 mg/kg) dose salvianolic acid B group, and salvianolic acid B+HIF-1 activation group, with 12 rats in each group. The bone structure at the joints of the rats in each group was measured and analyzed by micro-CT machine; The histopathological changes of cartilage were observed by safranin fast green staining and Masson staining; The contents of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and the activity of inducible nitric oxide syn- thase (iNOS) in rats were measured; The expression levels of HIF-1, VEGF, matrix metalloproteinase 13 (MMP-13) and activated caspase-3 (cleaved caspase-3) in rat cartilage tissue were measured by Western blot. Results Compared with the sham operation group, the cartilage tissue damage of the rats in the osteoarthritis group was severe, and the bone volume fraction (BV/TV) of the ankle bone was decreased, the ratio of ankle bone surface area to bone volume (BSA/BV), serum IL-6, TNF-α contents, iNOS activity, cartilage HIF-1, VEGF, MMP-13, and cleaved caspase-3 protein expression were increased (P<0.05); Compared with the osteoarthritis group, the cartilage tissue of the nimesulide group, the salvianolic acid B high and low dose salvianolic acid B group gradually recovered, and the ankle BV/TV was increased, BSA/BV, serum IL-6, TNF-α contents, iNOS activity, cartilage HIF-1, VEGF, MMP-13, and cleaved caspase-3 protein expression were decreased (P<0.05); Compared with the high-dose salvianolic acid B group, the salvianolic acid B+HIF-1 activation group had increased cartilage tissue injury, and ankle BV/TV was lower, BSA/BV, serum IL-6, TNF-α contents, iNOS activity, cartilage HIF-1, VEGF, MMP-13, and cleaved caspase-3 protein expression were increased (P<0.05). Conclusions Salvianolic acid B can inhibit the expression of HIF-1 and VEGF and relieve osteoarthritis by regulating the HIF-1/VEGF pathway.
    Clinical Sciences
    Aescuven forte improves the symptoms after treatment of Er:YAG laser-induced skin swelling in facial acne scars patients
    DING Wenyun , WANG Chenyu, WANG Wenqian, LONG Xiao
    2023, 43(1):  165-168.  doi:10.16352/j.issn.1001-6325.2023.01.0165
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    Objective To observe the efficacy of Aescuven forte in mitigating the skin turgor after Er:YAG laser treatment. Methods A total number of 64 patients who underwent twice 2 940 nm Er:YAG laser dot-matrix therapy was included in this study. Patients were randomly divided into control group and experimental group (oral administration of Aescuven forte 300 mg bid after therapy) at the first time of laser treatment. Follow-up was performed for a week after treatment and skin swelling was scored. Results The extent of skin swelling in experimental group was significantly lower than the control group on the day 1,2,3 and 5 after laser treatment(P<0.001). Skin swelling was remarkably subsided on day 6 in both groups and showed no difference between two groups. Conclusions Aescuven forte can significantly reduce skin swelling after 2 940 nm Er:YAG laser dot-matrix therapy in patients with facial acne scars.
    Mini Reviews
    Research progress of targeting on Nrf2/ARE pathway in the treatment of ischemic stroke
    YANG Xiaofei, LI Qiang, LI Yanfeng, WANG Hongquan
    2023, 43(1):  169-173.  doi:10.16352/j.issn.1001-6325.2023.01.0169
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    Recently, mounting evidences have suggested the therapeutic potential of Nrf2/ARE activators for treating Ischemic stroke. Moreover, several Nrf2 activators attenuate ischemic stroke-induced brain injury in several transient focal cerebral ischemia (tMCAO) in experimental animal models.
    Ferroptosis in the treatment of colorectal cancer
    ZHUANG Zhicheng, GUAN Guoxian
    2023, 43(1):  174-177.  doi:10.16352/j.issn.1001-6325.2023.01.0174
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    Ferroptosis is a form of programmed cell death caused by accumulation of iron-dependent lipophilic reactive oxygen species. Its occurrence depends on the comprehensive changes of intracellular iron homeostasis, lipid metabolism and antioxidant system. Colorectal cancer is one of the common malignant tumors of the digestive tract. It can effectively inhibit the proliferation of colorectal cancer cells, reverse drug resistance and improve the radio sensitivity of colorectal cancer cells by inducing ferroptosis pathway.
    Research progress of pathogenesis and treatment of microorganism-related tumors
    SUN Hui, CHEN Siyu
    2023, 43(1):  178-182.  doi:10.16352/j.issn.1001-6325.2023.01.0178
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    Tumor is a disease with multi-factor participation and multi-stage onset, among which the biological carcinogenic factors are mainly carcinogenic viruses, bacteria and parasites, and the common ones include hepatitis B virus(HPV) and hepatocellular carcinoma, Epstein-Barr virus(EBV) and nasopharyngeal carcinoma, human papilloma virus(HPV) and cervical cancer, Helicobacter pylori and MALT lymphoma,gastric cancer etc.The occurrence and development of certain tumors are closely related to microorganisms, which can play a role in carcinogenesis and cancer promotion through the following mechanisms:producing toxic substances, inducing toxic effects,inducing inflammation, affecting immune response, etc. This review briefly summarizes the carcinogenic and cancer-promoting mechanisms of microorganisms, and looks forward to the new ideas of using microorganisms as biomarkers for early tumor screening and developing drugs by targeting microorganisms.
    Research progress of antithrombotic drugs
    ZHANG Jie, MA Xuzhou, LI Huihui, WANG Miao
    2023, 43(1):  183-187.  doi:10.16352/j.issn.1001-6325.2023.01.0183
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    Thrombosis is the main direct cause of acute manifestations of atherosclerotic cardiovascular disease (myocardial infarction, stroke). Globally, the incidence of thromboembolic diseases has increased in recent years, accompanied by an increase in patient mortality. Currently,several targeted drug delivery strategies have been developed for thromboembolic diseases, including antiplatelet drugs and anticoagulant drugs. With the development of antithrombotic drugs, the treatment of cardiovascular diseases caused by thrombus shows significant potential benefit for patients.
    Research progress of gut microbiota in neuropathic pain
    LYU Yanhan, YU Zhuoying, LI Min
    2023, 43(1):  188-191.  doi:10.16352/j.issn.1001-6325.2023.01.0188
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    The mechanism of neuropathic pain (NP) is extremely complicated, and current pharmacological treatments for NP are still unsatisfactory. The gut microbiota plays important roles in inducing NP. The changes of gut microbiota and gut microbiota-dependent metabolite could promote the immune cells and glial cells releasing pro-inflammatory cytokines, initiating the innate immune response and inflammation by activating Toll-like receptors (TLRs), which is believed to be a potential candidate to alleviate NP.
    Research advances in the treatment of lung diseases based on mesenchymal stem cell exosomes
    CHEN Lili,SUN Yonghong,LEI Xiaoyan,WANG Jianjun,CHEN Xingxing,BAO Baixin
    2023, 43(1):  192-195.  doi:10.16352/j.issn.1001-6325.2023.01.0192
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    Exosomes secreted by mesenchymal stem cell(MSC-EXOs)carry lipids,proteins,nucleic acids and other signaling substances, which play an important role in intercellular communication, immune regulation and vascular regeneration. MSC-EXOs in pulmonary hypertension, pulmonary fibrosis and other lung diseases can improve the function of the lung tissue and treat lung diseases by regulating different signaling pathways, reducing cell inflammatory reaction, regulating immune function, improving the function of mitochondria, promoting angiogenesis, reducing pulmonary vascular remodeling and etc.,which provides new ideas for the treatment of refractory lung diseases in the future.
    Progress on application of dipeptidyl peptidase Ⅲ in the treatment of sepsis
    WANG Yingchen, SUN Liqun
    2023, 43(1):  196-199.  doi:10.16352/j.issn.1001-6325.2023.01.0196
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    Sepsis is still one of the important challenges of medicine in the 21st century. Dipeptide peptidase Ⅲ(DPP3)is a new biomarker associated with sepsis, which mainly degrades angiotensin Ⅱ in human body. It can not only inhibit cardiac systolic function, but also reduce vascular tension, resulting in insufficient cardiac output and low blood pressure, resulting in shock and multiple organ dysfunctions in patients with sepsis. Therefore, DPP3 may become a new target for sepsis treatment.
    Medical Education
    Role of bedside intensive video laryngoscope tracheal intubation training in emergency difficult airway intubation teaching
    LIU Anlei, LI Yan, ZHU Huadong, LIU Jihai, LI Yi, XU Jun, YANG Jing
    2023, 43(1):  200-203.  doi:10.16352/j.issn.1001-6325.2023.01.0200
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    Objective To explore the role of bedside intensive video laryngoscope tracheal intubation training in emergency difficult airway intubation teaching. Methods Eighty-four rotating physicians in emergency resuscitation room were randomly divided into a simulation training group and a bedside intensive training group with 42 in each. They were asked to fill the questionnaire before and after the training, complete the simulated video laryngoscope tracheal intubation and the patient video laryngoscope tracheal intubation independently. The training satisfaction, intubation willingness, expected intubation success rate, counting number of intubation attempts, and intubation success rate, total time length of intubation, evaluation of difficult airway, evaluation after intubation, qualified rate of catheter position, qualified rate of balloon pressure, incidence of adverse reactions and other data were collected. Results There were no significant difference between age and educational background from the simulation training group and the bedside intensive training group. The scores of training satisfaction (98.0±1.08) of trainees in bedside intensive training were higher than those of the simulation training group (94.7±2.7) (P<0.01); The scores of willingness to try difficult intubation after training in the bedside intensive group (8.6±0.7) were higher than those of the simulation training group scored (6.62±1.34) (P<0.01); The expected intubation success rate in the bedside strengthening training group was 82.0%±5.6%, which was higher than that in the simulation training group 60.3%±11.3%(P<0.01). In the evaluation of the intubation process, compared with the simulation training group, the bedside intensive training group had fewer intubation attempts [1.0±0.4 vs. 2.0±0.7], and the intubation success rate was higher [39(92.9) vs. 28(66.7) ], the total intubation time was shorter [38.8±3.3 vs. 50.5±5.6](both P<0.01). In terms of post-intubation evaluation, compared with the simulation training group, the bedside intensive training group had a higher qualified rate of balloon pressure [41(97.6%) vs. 35 (83.3%)] and a higher qualified rate of catheter position [41(97.6%)] vs. 35(83.3%)], fewer adverse reactions of intubation: cough [1(2.1%) vs. 8(19.1%)], throat injury [0(0.0%) vs. 6(14.3%)](all P<0.05). Conclusions Bedside intensive laryngoscope tracheal intubation training can improve emergency physicians' confidence and skill in handling difficult airways.
    Research on the hybrid teaching mode in the obstetrics and gynecology itinerant teaching of undergraduates
    SHI Jinghua, LI Xiaoyan, CHEN Rong, LUO Min
    2023, 43(1):  204-206.  doi:10.16352/j.issn.1001-6325.2023.01.0204
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    Objective To explore the effect of small private online course (SPOC) combined with flipped classroom (FCR) hybrid teaching model in itinerant teaching of obstetrics and gynecology for undergraduates. Methods For the obstetrics and gynecology interns of our Peking Union Medical College, the teaching group carried out blended teaching of SPOC and FCR. Examing scores were compared among students trained by the traditional teaching mode and by the online teaching mode. Results The scores of students from online teaching group in written examination, interview and total scores were significantly lower than those of traditional teaching mode. After the reform of online and offline teaching mode, the interview performance was significantly higher than that of the pure online teaching group. Conclusions The hybrid teaching model of SPOC combined with FCR may meet the needs of prevention and control measures in the post-COVID-19 pandemic era and improves students' academic record to a certain extent.
    Value of lectures on the standardization of image report writing in residency training of medical imaging
    CHE Shunan, YE Feng, ZHANG Yongming, ZHAO Xinming, ZHOU Chunwu
    2023, 43(1):  207-210.  doi:10.16352/j.issn.1001-6325.2023.01.0207
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    Objective To evaluate training course on the standardization of image report writing in medical imaging residency training in order to improve the uniformity of image report writing and report quality. Methods Residents who attended medical imaging training facility for study between September 2020 and September 2021 were divided into experimental and control groups according to whether or not they received lectures on imaging report standards. Tencent quiz software was used to evaluate the mastery of imaging report writing standards by two groups of residents. Results The results of resident self-assessment showed that the residents in the experimental group improved significantly in terms of reporting format specification, description of lesion-related secondary signs, report conclusion, report content, and total score of reporting standard after 6 months of training(P<0.05). The results of instructor evaluation showed that the experimental group had better reporting standard and quality evaluation indexes than the control group (P<0.05). Conclusions The lecture on image report writing standard has a positive impact on the mastering of image report writing standard and the promotion of report quality by medical imaging training residents.