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Table of Content

    05 June 2024, Volume 44 Issue 6
    Special Issues:Pathogenesis of Adrenal Tumors
    Advances in the pathogenesis of adrenal tumors
    TONG Anli
    2024, 44(6):  741-741.  doi:10.16352/j.issn.1001-6325.2024.06.0741
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    Immune microenvironment of pheochromocytomas and paragangliomas
    ZHOU Yue, TONG Anli
    2024, 44(6):  742-747.  doi:10.16352/j.issn.1001-6325.2024.06.0742
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    Pheochromocytomas/Paragangliomas(PPGLs) are rare neuroendocrine tumors with a high hereditary predisposition. The pseudo-hypoxic PPGLs dominated by SDHx mutations show a higher potential of metastasis. Approximately 10%-17% of PPGLs develop metastasis and patients with metastatic diseases have restricted treatment options and a poor prognosis. Tumor immune microenvironment (TIME) plays a crucial role in the development and progression of tumors as well as predicting patients’ prognosis and their response to immune checkpoint inhibitors. Several studies have initially characterized the immune landscape of PPGLs. This review focuses on the infiltration of immune cells,the expression of immune checkpoints in the TIME of PPGLs and their relationships with genetics and metastasis of tumors in order to better understand the mechanisms of tumor immune evasion in PPGLs and provide insights to support novel treatment strategy for metastatic PPGLs.
    Genetic and molecular mechanism changes of adrenocortical carcinoma
    LIU Jinghua, LU Lin
    2024, 44(6):  748-752.  doi:10.16352/j.issn.1001-6325.2024.06.0748
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    Adreno-cortical carcinoma (ACC) is an uncommon tumor, some of them may produce excessive steroid hormone. The prognosis of ACC is quite poor and current treatment options are relatively limited, especially for patients whose tumor is metastatic and difficult to remove by surgical operation. This makes the management of ACC challenging. Existing studies have shown that the mechanisms of ACC are complex. The molecular changes happen in adrenocortical development and homeostasis may explain the occurrence of ACC. This review focuses on recent studies and summarizes the genetic and molecular mechanism changes that potentially associated with ACC in the development process of the adrenal cortex and multi-omics studies, including genomic changes, epigenetic changes, differences in non-coding RNA, immune microenvironment and features found through multi-omics analysis. This information may support screening of therapeutic targets of ACC in future.
    Electrical excitability and aldosterone secretion in the zona glomerulosa cells:the role of ion channels
    ZHANG Xuefeng, HU Changlong
    2024, 44(6):  753-757.  doi:10.16352/j.issn.1001-6325.2024.06.0753
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    Aldosterone is a kind of steroid hormone produced by the zona glomerulosa (ZG) cells of the adrenal cortex which plays a crucial regulatory role in fluid-electrolyte balance and blood pressure stability. Aldosterone biosynthesis is a calcium-dependent process, and ion channels play a key role in this process. Primary aldosteronism is closely related to mutations in ion channels. Recent studies have highlighted an intriguing relationship between the unique rosette structure of ZG cells and their electrical excitability. Elucidating the connection between rosettes and ZG cell electrical excitability as well as aldosterone secretion leads to a deeper understanding of the mechanisms regulating aldosterone secretion and may provide new insights into the pathogenesis of primary aldosteronism.
    Mechanisms of cell death and proliferation of aldosterone-producing adenoma
    GAO Yinjie, TONG Anli
    2024, 44(6):  758-762.  doi:10.16352/j.issn.1001-6325.2024.06.0758
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    Aldosterone-producing adenoma (APA) is one of the main causes of primary aldosteronism (PA). Current researches on its autonomous secretion of aldosterone mainly focus on the detection of somatic driver genetic mutations. However, the specific mechanisms underlying the death and proliferation of adrenal cortical cells and the occurrence and development of adenomas are still unclear. This study aims to review the research progress on the mechanisms of cell death and proliferation of APA by combining previously published studies on genomics, transcriptomics, metabolomics, and epigenetics related to adrenal tissues.
    Original Articles
    Therapeutic effect of CAR-γδT cells targeting at BCMA in multiple myeloma
    LI Yinghui, XU Yi, ZHANG Jianmin, CHEN Hui, HE Wei
    2024, 44(6):  763-771.  doi:10.16352/j.issn.1001-6325.2024.06.0763
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    Objective To construct chimeric antigen receptors modified γδT cells targeting at BCMA (BCMA CAR-γδT) and to evaluate its efficacy of anti-multiple myeloma in vitro. Methods Lentiviral vectors containing BCMA single-chain variable fragment were constructed and transiently transfected into 293T cells. The expression of foreign genes was verified by fluorescence microscopy and Western blot; the lentivirus was packaged and the virus titer was determined by flow cytometry. Human peripheral blood αβT cells were infected and γδT cells were examined for its infection efficiency; LDH release method was used to detect the cytotoxic activity of BCMA CAR-γδT cells against human multiple myeloma cell lines in vitro, and the difference of cytotoxic activity between CAR-γδT cells and CAR-αβT cells was compared by Incucyte S3 Live-Cell Analysis Instrument. Results Twenty-four hours after BCMA-CAR lentiviral vector was transferred into 293T cells, the expression of exogenous ZsGreen was microscopied by fluorescence microscope; CD3ζ was detected by Western blot, which showed that BCMA-CAR could be successfully expressed. The lentivirus was packaged, collected and concentrated(virus titer of 2.23×108 Tu/mL). Infected αβT cells and γδT cells from human peripheral blood in MOI=5, and the results of flow cytometry showed that infection efficiency of αβT cells was 59.18%±2.56%, γδT cells was 48.15±9.86%. The cytotoxic activity of CAR-γδT cells against human myeloma cell lines MM1.S, H929 with high expression of BCMA and K562 cells with over-expression of BCMA was higher than that of empty vector control γδ T cells,which were significantly enhanced (P<0.001), but there was no difference in cell lines negative for BCMA expression; Live-Cell Analysis Instrument results showed that the cytotoxic activity of BCMA CAR-γδT cells and BCMA CAR-αβT cells against H929 in vitro was significantly better than their vector control cells. There was no difference in the cytotoxic activities of BCMA CAR-γδT cells as compared with against BCMA negative cell lines, and so do BCMA CAR-T cells. Conclusions Cytotoxic activity of BCMA CAR-γδT targeting at BCMA in vitro was significantly enhanced ,which is expected to serve as a novel allogeneic γδT cell product for cell adoptive immunotherapy of multiple myeloma.
    Bone marrow-derived mesenchymal stem cells relieve the adverse effects of simulated microgravity on mouse cerebral cortex
    GONG Jintao, LI Jianwei, LI Yuheng, LI Qian, ZHAO Chunhua
    2024, 44(6):  772-778.  doi:10.16352/j.issn.1001-6325.2024.06.0772
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    Objective To explore the protective effects and the underlying mechanisms of bone marrow mesenchymal stem cells (BMSCs) on the brain in microgravity environment. Methods The physiological effects of microgravity were simulated with the hindlimb unloading (HU) mouse model. The animals were divided into 3 groups: the control group (wild-type mice), the hindlimb unloading (HU) group, and the BMSCs treatment group, with six in each. RT-qPCR was adopted to determine the expression levels of pro-inflammatory cytokines (Il-6, Il-1β and Tnf-α) in the cerebral cortex; immunohistochemistry was performed to evaluate the proportions of microglia (IBA-positive) and astrocytes (GFAP-positive); Western blot was used to measure the expression levels of apoptosis-and senescence-related molecules (BAX, BCL-2, p21, and p53). Results Compared with wild-type mice, the expression levels of Il-6, Il-1β and Tnf-α in the cerebral cortex of HU mice were significantly increased (P<0.05), the expression level of BAX, p21 and p53 was also significantly increased(P<0.05). However, the expression level of BCL-2 protein were significantly decreased (P<0.05). The proportion of microglia (IBA1 positive) and astrocytes (GFAP positive) was increased (P<0.05); After BMSCs treatment, the expression level of Il-1β in the cerebral cortex of HU mice was significantly decreased (P<0.05), the expression levels of BAX, p21 and p53 were significantly decreased (P<0.05), and the expression level of BCL-2 protein was significantly increased (P<0.05). The proportion of microglia (IBA1 positive) and astrocytes (GFAP positive) decreased (P<0.05). Conclusions BMSCs potentially relieve the detrimental effects of simulated microgravity on mouse brain by inhibiting inflammation, apoptosis and cellular senescence.
    Dynamic transcriptomic analysis of macrophages infected with Salmonella typhimurium
    SONG Boyuan, WU Xueli, LI Xueyuan, WANG Lisha, CHEN Yang
    2024, 44(6):  779-785.  doi:10.16352/j.issn.1001-6325.2024.06.0779
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    Objective To comprehensively understand the dynamic transcriptional landscape during infection through investigating the temporal molecular changes in macrophages RAW 264.7 upon infection with Salmonella typhimurium SL1344. Methods Macrophages RAW 264.7 were infected with Salmonella typhimurium SL1344, and cell samples were collected at 0 h, 8 h, and 16 h for RNA-sequencing(RNA-seq). Upstream and downstream analyses of the transcriptome data including differential gene expression, clustering, functional annotation, and molecular network studies were conducted to elucidate the signaling pathways changes in macrophages. Results Infected macrophages exhibited significant morphological and transcriptional changes. Differential gene analysis identified significant upregulation and downregulation patterns. Clustering revealed six gene clusters involving various signaling pathways, such as immune response, membrane transport, and lipid catabolic process. Conclusions Macrophages dynamically respond to Salmonella typhimurium infection, displaying distinct temporal gene expression patterns. The coordinated activation of immune response, membrane transport, and lipid catabolic process pathways implies a multifaceted cellular adaptation to external infections, providing essential insights into the molecular mechanisms of macrophage response to Salmonella typhimurium infection.
    Small molecule compounds improve the targeted differentiation efficiency of cerebral organoids from pluripotent stem cells
    ZHU Wanwan, ZHOU Jinjuan, XIU Jianbo
    2024, 44(6):  786-792.  doi:10.16352/j.issn.1001-6325.2024.06.0786
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    Objective To optimize the conditions and improve the efficiency for the differentiation of pluripotent stem cells (PSCs) into cerebral organoids. Methods Based on the induction differentiation system for inducing human embryonic stem cells (hESCs) H9 towards cerebral organoids, a combination of small molecule compounds was added to the developmental stage of neural progenitor cells in the early stage of cerebral organoid differentiation, and the efficiency of neural progenitor cell formation, apoptosis and differentiation into neurons in cerebral organoids in the differentiation stage were observed through morphology, the expression of marker genes was detected by RT-qPCR and the effect of small molecule compound combination on cerebral organoids was comprehensively evaluated. Results At the critical stage of neural progenitor cell development (1 d-14 d) of cerebral organoids, dorsomorphine, A83-01, GSK-3β inhibitor CHIR99021 and SMAD inhibitor SB-431542 were successively added to the medium. It could significantly increase the expression of marker genes in the neural progenitor cell stage, promote the formation of specific neural tube-like structures and reduce apoptosis in the central region of cerebral organoids. Conclusions By using the combination of four small molecule compounds, the formation efficiency in early cerebral organoids can be significantly improved, apoptosis in cerebral organoids can be reduced, neuronal formation can be promoted, and tissue structure heterogeneity in the culture process can be reduced.
    Effects of intraperitoneal injection of busulfan on metabolic characteristics of spermatogonial stem cells
    YU Zhixin, MANG Xinyu, ZOU Dingfeng, MIAO Shiying, SONG Wei, LI Kai
    2024, 44(6):  793-799.  doi:10.16352/j.issn.1001-6325.2024.06.0793
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    Objective To establish a mouse model treated with busulfan and to investigate its effects on the metabolism of spermatogonial stem cells (SSCs) of mouse testis. Methods C57BL/6J male mice with age of 8 weeks were injected with 10 mg/kg of busulfan intraperitoneally, then Thy1 positive cells were selected by immunomagnetic beads on day 0, day 5 and day 10 and followed by identification for purity and metabolomic analysis. ResultsThe testis weight ratio decreased and the tissue structure of testis was damaged (P<0.05). Based on the results of principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) , there were significant metabolic differences between the sample groups treated for 0 d, 5 d and 10 d. A total of 89 differential metabolites were identified including glutathione (GSH), arginine and unsaturatedfatty acids (UFAs), and their important metabolic pathways involved glycerophospholipid metabolism, arginine and proline metabolism. Conclusions Affecting the specific metabolic pathway may result in obvious reproductive toxicity and lead to decrease of testicular weight as well as tissue structure damage in mice. Metabolomic analysis showed that the potential reproductive toxicity mechanism of SSCs may be related to the metabolic pathways such as lipid metabolism, arginine and proline metabolism.
    Bioinformatics analysis of differential expression of CD44 in glioblastomas and cell experimental validation
    SUN Xu, LI Shunshun, WANG Dianheng, WANG Zhenfeng
    2024, 44(6):  800-808.  doi:10.16352/j.issn.1001-6325.2024.06.0800
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    Objective To investigate the clinical significance of the differential expression of CD44 in glioblastoma (GBM), and the enrichment of related pathways combined with U87 cell verification. Methods Differential expression and Cox analysis were used to find potential differences in CD44 expression between tumor patients and healthy people as shown by pancancer transcriptome and hazard ratio (HR). Immunoinfiltration and stem-cell related scores of GBM patients with high and low CD44 expression groups were compared and the differences between immune cell subsets, and their correlation with CD44 were further analyzed. Pathway enrichment of differentially expressed genes in high and low CD44 expression groups for GBM patients was performed. Over-expression (OE), knockdown (KD) and knockout (KO) of CD44 in U87 cells was done by constructing and packaging lentivirus and using the electroribonucleoprotein complex; CD44 immunofluorescence staining was performed on U87 and U87 CD44 OE cells. The activity and apoptosis of U87 cells were detected by knocking down CD44 and the migration and invasion ability of U87 cells were detected by knocking down CD44. Results The expression of CD44 in GBM patients was higher than that in healthy people (P<0.05) and HR>1. GBM patients with high CD44 expression had higher stromal cell and immunoinfiltration scores, and GBM patients with high and low CD44 expression had significant differences in the ratio of dendritic cells, CD4+ memory T cells and regulatory T cells, all positively correlated with CD44 expression (P<0.05). Differential gene enrichment in GBM patients with high and low CD44 expression was further associated with pathways related to cell migration and apoptosis(P<0.05). Experiments using U87 cells showed that CD44 was normally localized in the cell membrane, but for CD44 OE it accumulated in the cytoplasm. CD44 KD can lead to a decrease in cell viability, increased in cell apoptosis, with the cell migration and invasion ability of CD44 KO also decreasing (P<0.05). Conclusions Low expression of CD44 can decrease viability, migration and invasion of U87 cells and promote apoptosis rate of U87 cells, which leads to the deterioration of GBM and is a related factor potentially affecting the prognosis of GBM patients.
    Venlafaxine stabilizes axons of the neurons in depression model mice
    SUI Jiaping, WEI Hui
    2024, 44(6):  809-815.  doi:10.16352/j.issn.1001-6325.2024.06.0809
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    Objective To investigate whether the effects of venlafaxine on major depression disorder is associated with ankyrin G. Methods Breed Synapsin-Cre1 and Ankyrin3-floxed mice (Ank3 cKO mice). Ank3 cKO mice and wild type mice were randomly divided into model and control groups. All mice in model group and control group were orally administrated with venlafaxine(1 g/L) or the solvent(normal saline, NS) with the volume of 200 μL,respectively. Depression-related behaviors were examined by sucrose preference test (SPT) and Y maze test. The level of ankyrin G and PSD95 in the cortex of four groups were detected by Western blot. The level of ankyrin G and MAP2 in the in hippocampus of four groups were detected by immunohistochemistry method. ResultsCompared with wt-saline group, the cKO mice in saline showed a significantly decreased preference of sucrose (P<0.001) and low spontaneous alteration(P<0.05). Compared with cKO control ones, the venlafaxine model cKO mice showed remarkably increased preference of sucrose(P<0.001) and more spontaneous alteration(P<0.05). The level of ankyrin G and PSD-95 in the cortex of venlafaxine cKO mice was much higher than that in control mice(P<0.01)The level of ankyrin G and MAP2 in the hippocampus of venlafaxine cKO mice were much higher than those of control mice(P<0.05). Conclusions Venlafaxine alleviates the depression symptoms caused by knocking down Ank3. The mechanism of depression treatment by venlafaxine is potentially associated with levitating ankyrin G level in prefrontal cortex and hippocampus.
    Datura metel L. inhibits the secretion of inflammatory factors and angiogenesis in keratinocytes in vitro
    ZHANG Boping, SI Xinlei, WU Fenfang
    2024, 44(6):  816-820.  doi:10.16352/j.issn.1001-6325.2024.06.0816
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    Objective To investigate the potential role of the traditional herbal medicine Datura metel L. in the treatment of psoriasis using TNF-α-induced inflammation in keratinocytes as a model. Methods Keratinocyte cell line HaCaT was used to establish a psoriasis cell model by tumor necrosis factor-alpha (TNF-α) treatment. The experiment comprised three groups: a blank control group, TNF-α-induced psoriasis model group, and TNF-α + Datura metel L. intervention group. Level of IL-17 and CCL20 was measured ELISA, expression of nuclear factor kappa B (NF-κB) subunit p65 protein was measured by Western blot. Endothelial cell tube formation experiment was conducted using an in vitro angiogenesis analysis kit. Results Compared to the TNF-α-induced psoriasis model group, the Datura metel L. extract significantly reduced the levels of IL-17 and CCL20 in the cell culture supernatant of TNF-α-induced psoriasis model (P<0.001); Datura metel L. extract markedly decreased the NF-κB subunit p65 protein level in TNF-α-induced psoriasis model cells (P<0.01); Datura metel L. extract effectively inhibited the induction of endothelial cell tube formation by the cell culture supernatant of the psoriasis model group. Conclusions The Datura metel L. extract down-regulates NF-κB signaling pathway molecules, reducing the production of IL-17 and CCL20 inflammatory factors in inflammatory keratinocytes and inhibiting angiogenesis.
    Design and investigation of CRISPRi tools based on dCasMINI protein
    CHEN Xinwen, CAO Jiaxuan, RAO Shuquan
    2024, 44(6):  821-827.  doi:10.16352/j.issn.1001-6325.2024.06.0821
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    Objective To explore the design of CRISPR interference (CRISPRi) tools based on the deactivated CasMINI (dCasMINI) protein and to evaluate their transcriptional inhibition effects. Methods The tetracycline-on (tet-on) system, flow cytometry, and quantitative reverse transcription polymerase chain reaction (RT-qPCR) were used to evaluate the transcriptional inhibition effects of dCasMINI system in mammalian cells at three level-plasmid genes, exogenous genomic loci, and endogenous genomic loci. Additionally, six dCasMINI-CRISPRi tools (dCasMINI, dCasMINI-ZIM3 KRAB, dCasMINI-KRAB-MeCP2, dCasMINI-ZNF324 KRAB, dCasMINI-3x KRAB, and dCasMINI-Com-KRAB-MECP2) were designed and compared for their transcriptional inhibition effects along with single guide RNA (sgRNA) at different positions. Results dCasMINI, dCasMINI-ZIM3 KRAB,dCasMINI-KRAB-MeCP2, dCasMINI-ZNF324 KRAB, dCasMINI-3x KRAB, and dCasMINI-Com-KRAB-MeCP2 exhibited varying degrees of transcriptional inhibition on plasmids genes and exogenous genomic genes (P<0.05). Additionally, dCasMINI-ZIM3 KRAB, dCasMINI-KRAB-MeCP2, dCasMINI-ZNF324 KRAB, and dCasMINI-Com-KRAB-MeCP2 demonstrated different levels of transcriptional inhibition on endogenous genes (P<0.05). Different positions of sgRNAs showed distinct transcriptional inhibition effects (P<0.05). Conclusions The CasMINI system can be adapted into various CRISPRi tools for gene knockdown studies, with potential applications in various scenarios such as epigenetic gene editing in primary cells, in vivo screening, and clinical therapy in the future.
    Change of cell cycle in rat spleen after severe abdominal infection
    LI Jinping, LIU Hanqing, YANG Quanhui
    2024, 44(6):  828-832.  doi:10.16352/j.issn.1001-6325.2024.06.0828
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    Objective To explore the mortality rate and changes in splenic cell cycle and apoptosis in rats with abdominal infection. Methods An animal model of sepsis was induced by cecal ligation and puncture (CLP) in rats. At serial time points 0 h, 6 h, 12 h, 24 h, 48 h, and 72 h after CLP, the animal death rate was calculated. The percentage of cell cycle G1, S, G2/M phase and apoptosis of isolated spleen cells were analyzed using flow cytometer. The expression of p27 of spleen tissue was determined by Western blot analysis. Results In the rat model of CLP-induced systemic inflammatory response, the mortality of animals gradually increased at 0 h, 6 h, 12 h, 24 h, 48 h and 72 h after CLP surgery, reaching a maximum of 88.81%. The abdominal infection in the animals gradually worsened at 0 h, 6 h, 12 h, and 24 h after CLP surgery, but localized and gradually alleviated after 48 h and 72 h. Within 48 h after CLP surgery, there was a blockage in the G1 phase of the spleen cell cycle, an increase in the percentage of apoptotic cells, and an elevation in p27 expression. After 48 h, the G1 phase blockage gradually recovered, the number of apoptotic cell decreased, and p27 expression declined. Additionally, within 48 h after CLP surgery, there was a decrease in the percentage of cells in the S and G2/M phases of the spleen cell cycle, but the percentage of cells in these phases gradually increased after 48 h. ConclusionsCell cycle arrest and apoptosis of the spleen cells are potentially contributed to the change of animal mortality after abdominal infection.
    Association between daunorubicin resistance and PD-L1 protein expression in children with acute myeloid leukemia
    SONG Lili, GUAN Yujie, MA Ping, LI Ning
    2024, 44(6):  833-839.  doi:10.16352/j.issn.1001-6325.2024.06.0833
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    Objective To explore the correlation between daunorubicin (DNR) resistance and expression of programmed death receptor ligand 1 (PD-L1) in children with acute myeloid leukemia (AML). Methods Totally 110 bone marrow samples were selected from AML patients admitted to Zhengzhou University Affiliated Children's Hospital from January 2016 to December 2022 as the study group, 50 bone marrow samples from normal bone marrow donors were used as the control group. Human AML cell lines HL60, THP-1, U-937, and Molm-13 were cultured, and the expression of PD-L1 protein was detected by Western blot. LV-PD-L1-shRNA and LV-PD-L1-WT-OE lentiviral vectors were constructed. The effect and mechanism of PD-L1 on DNR resistance in Molm-13 cells were analyzed. Results The expression level of PD-L1 protein was higher than that in control group and the expression level of PD-L1 protein in AMI cell lines was higher than that in healthy bone marrow mononuclear cells (BMMC) (P<0.05). PD-L1 expression was related to white blood cell count, bone marrow primitive cell ratio, prognostic risk stratification and disease remission after two standard chemotherapy regimens in AML patients (P<0.05). The overall survival rate of the PD-L1 high expression group was lower than that of the PD-L1 low expression group (P<0.05). Compared with the LV-PD-L1-WT-OE group, the LV-PD-L1-shRNA group showed a decrease in PD-L1 mRNA expression, cell proliferation activity but an increased apoptosis rate(P<0.05). LV-PD-L1-shRNA enhanced sensitivity of Molm-13 cells to DNR. TCGA database analysis showed that glucose 6-phosphate dehydrogenase (G6PD) was a potential target gene for PD-L1. Conclusions The high expression of PD-L1 in pediatric AML is related to chemotherapy resistance in children with AML, and DNR resistance might be caused through regulation of G6PD.
    Clinical Sciences
    Efficacy of ozone combined with low temperature plasma radiofrequency ablation in the treatment of cervical spondylotic radiculopathy
    HUO Yansong, SUN Haiyan, PANG Jinlei, GUO Xiangfei, LIU Yajing, RAN Guangyuan, HE Mingwei
    2024, 44(6):  840-844.  doi:10.16352/j.issn.1001-6325.2024.06.0840
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    Objective To investigate the therapeutic effectiveness of ozone combined with low-temperature plasma coagulation therapy on patients with cervical spondylotic radiculopathy and its influence on inflammatory responses. Methods Ozone in combination with low-temperature plasma radio-frequency coagulation was applied to 75 patients with cervical spondylotic radiculopathy in Pain Medicine Department of Capital Medical University Affiliated Beijing Anzhen Hospital from May 2022 to May 2023. Pain scores were assessed using Visual Analog Scale (VAS) and Neck Disability Index (NDI) before and two weeks after treatment. Enzyme-linked immunosorbent assay (ELISA) was used to analyze the level of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interferon-gamma (IFN-γ) before and two weeks after treatment. Results After treatment with ozone plus low-temperature plasma radiofrequency, VAS and NDI scores showed a significant decrease[VAS: 5.36(4, 7) vs. 1.32(1, 2), P<0.000 1; NDI: 32.72(24, 70) vs. 7.62(3.55, 8.9), P<0.000 1]. Two weeks after surgical intervention, there was an effective reduction in the level of IL-6, TNF-αand IFN-γ alleviating the inflammatory response [IL-6: 4.33(2.51, 5.04) vs. 3.49(2.08, 4.43), P<0.05; TNF-α: 1.95(1.41, 2.21) vs. 1.61(1.02, 2.03), P<0.05; IFN-γ: 1.84(1.18, 2.47) vs. 1.55(0.76, 2.09), P<0.05]. Conclusions This Ozone combined with low-temperature plasma radiofrequency ablation is an effective technology for treatment of cervical spondylotic radiculopathy.
    Heart failure prediction model based on machine learning algorithms
    HU Chuanli, HE Xiaosong, ZHAO Jiang, LI Hua
    2024, 44(6):  845-852.  doi:10.16352/j.issn.1001-6325.2024.06.0845
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    Objective To construct a model of heart failure risk prediction based on four machine learning algorithms in order to support early diagnosis and intervention. Methods After reviewing the heart failure dataset published on the Kaggle community, feature selection was used to select relevant factors related to heart failure as predictive indicators. Four machine learning algorithms, namely logistic regression, support vector machine, random forest, and XGBoost were selected to establish predictive models. Compared and analyzed its accuracy, precision, recall, F1 score and area under the ROC curve (AUC) to verify the performance of the model. Results The study analyzed 11 features of 918 patients with heart failure and selected 10 feature factors for modeling. After optimizing the hyper-parameters through grid search, the XGBoost model performed the best, with accuracy, precision, recall, and f1_score and AUC values were 87.5%, 90.38%, 89.71%, 90.04% and 0.93, respectively. In addition, data analysis showed that exercise ST slope, chest pain type, and exercise induced angina were main influencing factors for heart failure. Conclusions The XG Boost model has the best predictive tool for heart failure, and machine learning algorithms may support early prevention, early diagnosis as well as control of heart failure.
    Investigation and risk factors analysis of hypertensive retinopathy patients in Zhangjiakou city
    ZHANG Jia, WANG Yanli, SONG Xiaocong, WANG Shuzhen
    2024, 44(6):  853-857.  doi:10.16352/j.issn.1001-6325.2024.06.0853
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    Objective To investigate the situation and risk factors of hypertensive retinopathy in Zhangjiakou city. Methods The residents from 19 counties and districts in Zhangjiakou were screened for hypertension and blood glucose level. Blood pressure, age, gender, course of disease, body mass index (BMI) and complications of the patients were collected by a questionnaire survey. The prevalence of hypertensive retinopathy was analyzed and the risk factors affecting the patients were analyzed. Results A total of 1 320 hypertension patients were found in 8 056 residents with prevalence rate as 16.39% (1 320/8 056) and 212 of the hypertensive patients were found to have retinopathy.The prevalence rate of retinopathy was 13.06% (212/1 320) in hypertensive patients and 2.63% in all the examined residents. The proportion of patients aged ≥60 was higher than that of patients aged<60 years old, the proportion of patients without hypertension treatment history was higher than that of patients with hypertension treatment history (P<0.05). The disease course, systolic blood pressure, diastolic blood pressure, diabetes and smoking history in the patient group were higher than those in the control group(P<0.05). Long hypertension course of disease, high systolic and diastolic blood pressure and diabetes history were risk factors for the occurrence of hypertensive retinopathy (P<0.05). Conclusions The incidence hypertensive retinopathy in 19 counties and districts of Zhangjiakou city is low but is high among residents aged ≥60 years. The risk factors are long course of disease, high systolic and diastolic blood pressure and diabetes history.
    Techniques and Methods
    Direct TaqMan-PCR based technology for non-invasive genotyping of lipid metabolism associated SNPs
    SUN Yuanhong, XIE Lyu, FAN Lihua, ZHENG Zhi
    2024, 44(6):  858-865.  doi:10.16352/j.issn.1001-6325.2024.06.0858
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    Objective To establish a non-invasive single nucleotide polymorphism genotyping technology based on direct TaqMan-PCR for high-throughput genotyping of site rs688 and rs964184 associated with lipid metabolism to meet the need for early screening of at-risk populations. Methods Extracted DNA was used to optimize the PCR annealing extension temperature and amplification procedure for the designed TaqMan probe; the oral swabs were placed into the sample treatment solution and then briefly centrifuged, and a small amount of the supernatant was taken for the direct qPCR amplification analysis; the freeze-thawing stability of probe and the effects of sample storage time on genotyping results were explored. Results The technology requires only simple treatment of oral swab for PCR analysis and can be stored at room temperature for 4 d. The optimized method was able to distinguish homozygous wild type, homozygous mutant type and heterozygote at rs688 and rs964184 loci. Conclusions A fast, convenient, high-throughput, and low-cost SNP genotyping method has been established, providing an efficient and accurate new approach for large-scale screening of high-risk populations carrying susceptibility genes.
    Case Reports
    Beneficial effects of vemurafenib on craniopharyngioma carrying BRAF-V600E mutation
    WANG Xi, YE Ting, NIE Min, WU Xueyan, MAO Jiangfeng
    2024, 44(6):  866-872.  doi:10.16352/j.issn.1001-6325.2024.06.0866
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    Objective To evaluate the efficacy and adverse reactions of BRAF inhibitor vermorafenib on the treatment of refractory craniopharyngioma carrying BRAF-V600E mutation. Methods Clinical data of two patients with refractory craniopharyngiomas (CP) were recorded and reviewed. The patients were followed up for 3-5 years. Literature on CPs receiving BRAF or BRAF/MEK therapy was reviewed. Results 1)Papillary CP progressed after multiple operations and radiotherapy in two patients. Further treatments were very difficult. 2)The presence of BRAF-V600E mutation in the tumor was confirmed, and vermorafenib was administered for 6.5-7.5 months. Tumor volumes remarkably shrank by 95%-99%. No tumor relapse was observed during the follow-up of 3-5 years after discontinuation of vemurafenib. 3)The main adverse reaction was rash, which was dose dependent. 4)Literature review showed the volume shrank by 50%-100% in 33/34 tumors during BRAF or BRAF/MEK inhibitor therapy. Conclusions BRAF inhibitor vemurafenib is effective in treating refractory craniopharyngioma carrying BRAF-V600E mutation with endurable side effects, which may bring some changes to the management of CP in future.
    A case of visceral myopathy with ATCG2 gene mutation misdiagnosed as Hirschsprung disease
    LIU Yuhao, ZHANG Yueyi, BAI Xiaoyin, CHEN Yang, ZHOU Weixun, LI Xiaoqing
    2024, 44(6):  873-876.  doi:10.16352/j.issn.1001-6325.2024.06.0873
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    Objective To discuss the clinical features, differential diagnosis and complication treatment of a patient with genetic visceral myopathy. Methods Medical history, physical examination and laboratory results of the patient were collected in detail. The pathology of previous surgery was reviewed. The patient's peripheral blood DNA was extracted and submitted for whole-exome sequencing. Subsequent Sanger sequencing was used to complete the pedigree verification of the mutation site. Results The patient was a young female presented with repeated incomplete intestinal obstruction since early childhood. She used to be misdiagnosed as Hirschsprung's disease for a long period and underwent multiple gastrointestinal segment resections. Her intestinal obstruction symptoms were temporarily relieved by surgeries, but severe diarrhea, mucus and bloody stools and malnutrition gradually occurred after the last operation. The patient had bacterial overgrowth in small intestinal tract and followed by intestinal opportunistic infections secondary to chronic intestinal pseudo-obstruction. The symptoms improved after anti-infection and enteral element diet treatment. Further pathological consultation and whole-exome gene sequencing confirmed the diagnosis of visceral myopathy related to ATCG2 R148L mutation. Conclusions Patients with early onset of chronic intestinal pseudo-obstruction and have poor response to conventional treatment are recommended to perform genetic test. The patients with hereditary visceral myopathy are susceptible to opportunistic intestinal infection. Attentions should also be paid to the prevention and treatment of complications to avoid unnecessary surgery.
    Mini Reviews
    Mitophagy in hepatic ischemia-reperfusion injury
    ZHANG Xiangxin, LI Wen
    2024, 44(6):  877-881.  doi:10.16352/j.issn.1001-6325.2024.06.0877
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    In case of hepatic ischemia-reperfusion injury (HIRI), mitophagy is induced by multiple pathways such as PINK1/Parkin and mitophagy receptor, which is closely related to mitochondrial dynamic related protein. Mitophagy is a double-edged sword. Moderate mitophagy is effective in reducing the oxidative stress and maintaining mitochondria homeostasis, thereby reducing hepatocyte apoptosis and necrosis, inhibiting inflammation response and mitigating HIRI. However excessive mitophagy will aggravate HIRI. So research on the role and mechanism of mitophagy may facilitate the development of new strategy for prevention and clinical treatment of HIRI.
    Metabolic reprogramming in idiopathic pulmonary fibrosis
    DUAN Ran, LI Qingyuan, FENG Tong
    2024, 44(6):  882-886.  doi:10.16352/j.issn.1001-6325.2024.06.0882
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    Pulmonary fibrosis is caused by repeated damage to the pulmonary alveolar epithelium, leading to abnormal epithelial-mesenchymal transition and myofibroblast production, resulting in the accumulation of extra cellular matrix and remodeling of the interstitium. Analogous to many tumor cells, pulmonary fibrosis involves metabolic reprogramming, encompassing alterations in carbohydrate, lipid, and amino acid metabolism. Notably, this reprogramming is marked by enhanced glycolysis, diminished fatty acid oxidation paired with augmented synthesis, and increased degradation of glutamine. Glycolysis efficiently and rapidly fulfills the energy requirements for the proliferation of macrophages and fibroblasts during fibrotic development. Additionally, the reprogramming of amino acid metabolism in activated fibroblasts not only facilitates collagen synthesis but also intensifies myofibroblast activation by generating reactive oxygen species (ROS) during the production of hydroxyproline.
    Regulation of host immune function by gut microbiota-derived secondary bile acids
    YUE Lingling, WANG Zihui, LI Xiaoqin, LI Lifeng, ZHANG Wancun, YU Zhidan
    2024, 44(6):  887-891.  doi:10.16352/j.issn.1001-6325.2024.06.0887
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    Disturbances of gut microbiota may affect the balance of the host immune system. The metabolism of gut microbiota produces many bioactive molecules interacting with host, typically secondary bile acids (SBAs). SBAs are involved in regulating the energy metabolism and the expression of inflammatory response-related genes by binding to membrane receptors and nuclear receptors, such as takeda G protein-coupled receptor (TGR5) and farnesol X receptor (FXR), which are essential for maintaining host immune homeostasis.
    The CXCL10/CXCR3 axis in acute respiratory distress syndrome
    SHENG Qi, TONG Jin
    2024, 44(6):  892-896.  doi:10.16352/j.issn.1001-6325.2024.06.0892
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    The CXC chemokine ligand-10/CXC receptor-3 (CXCL10/CXCR3) axis, a crucial pathway mediating Th1-type immunity, substantially contributes to the development of acute respiratory distress syndrome(ARDS) by enhancing inflammatory storms. It additionally promotes vascular endothelial apoptosis and impedes vasculature repair, elevating the risk of pulmonary vascular thrombosis. Conversely, this axis plays a protective role in limiting extra-cellular matrix deposition and modulating cell migration, thereby mitigating pulmonary fibrosis. The various mechanisms of the CXCL10/CXCR3 axis in the pathogenesis and progression of ARDS could offer novel insights for the diagnosis and treatment of ARDS.
    Medical Education
    Data-driven educational transformation may improve digital literacy of faculty in medical college
    HUANG Fumin, YAN Hongyu, JIA Qiannan, GAO Xiaohui
    2024, 44(6):  897-900.  doi:10.16352/j.issn.1001-6325.2024.06.0897
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    With the progressive advance and application of information technology, general artificial intelligence technology is leading the fourth industrial revolution, and opens a new era in terms of changing people's production, life and learning. The digital age is characterized by more emphasis on the comprehensive cultivation of competence, trans-border integration, collaborative communication, innovative consciousness, critical thinking and problem-solving. Data-driven educational transformation is one of the important directions of education reform, in which digital literacy of college teachers is crucial to data-driven educational transformation. From the perspective of data-driven educational transformation enabling the improvement of digital literacy of medical college teachers, the present article analyzed the positive impact on improving the quality and personalization of education as well as professional capacity building and faculty development to meet the needs of medical education in the new era.
    Practice and thinking on the reform of medical functional experiment and the construction of functional laboratory at Peking Union Medical College
    YU Xiaoli, JI Chao
    2024, 44(6):  901-904.  doi:10.16352/j.issn.1001-6325.2024.06.0901
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    Functional experiment is an important part of training program of basic medical experiment technology in medical colleges, and a scientific laboratory management system is a prerequisite for ensuring the progress and training outcomes of the education. Based on the school-running concept of “small-scale elite education” in Peking Union Medical College, this paper expounds the progress of the construction of medical functional experiment teaching system in Peking Union Medical College from many aspects, such as the reform of medical functional experiment teaching, the construction of functional laboratory and so on, and puts forward some ideas for building of a qualified training laboratory, so as to provide reference for further deepening the reform of experimental teaching and supporting sustainable development of teaching laboratory of medical schools.