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  • Expert review
    Stress medicine： current situation and prospect
    JIANG Chunlei
    2023, 47(9): 641-645. https://doi.org/10.7644/j.issn.1674-9960.2023.09.001
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    The emergence of stress medicine is based on the fact that humans are subjected to increasingly intense physical and psychological stress and stress-induced or exacerbated stress diseases. Stress medicine is a comprehensive interdisciplinary subject that studies the coping responses to stressors such as special environment, occupational factors and demands, as well as related medical problems. With the focus of stress research gradually shifting from physical stress to psychological stress and the increasingly close relationship between stress and diseases, stress medicine gradually emphasizes psychological stress, highlights special stress, improves operational ability, and highlights medical elements. By studying the biological mechanisms that trigger mental symptoms, developing new diagnostic criteria and treatment methods based on genetic, physiological, and cognitive data rather than just symptoms, is a trend in the prevention and treatment of stress diseases.
    • Original articles
  • Original articles
    Dietary nutrition and health status of personnel of the Armed Police
    ZHANG Yuxian, XIE Jinli, BI Xun, WANG Feng, YAO Zhanxin
    2023, 47(9): 646-649. https://doi.org/10.7644/j.issn.1674-9960.2023.09.002
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    Objective To investigate the dietary nutrition and health status of new recruits and personnel for special operations in the Armed Police,and to provide data for the formulation of personalized nutrition strategies for these populations. Methods A 3-day dietary survey was conducted using the weighing method. The 251 recruits and 226 special operations personnel were all male. The body mass index （BMI） and body fat rate of 1264 recruits were measured with a body composition analyzer. Results Only the intakes of livestock meat,poultry meat,poultry eggs,milk and fruits met the current military standard in the recruit brigade. The energy supply ratio of the three major nutrients was basically balanced. Calcium,zinc,vitamin A,vitamin B1 and vitamin B2 did not meet the requirements of the current military standard. About 20.96% and 1.90% of the recruits were overweight and underweight respectively. Only the intakes of livestock meat,poultry eggs,milk and fruits met the requirements of the current military standard in this special operations brigade. The energy supply ratio of the three major nutrients was unbalanced. The intakes of vitamin A and vitamin B1 fell short of the current military standard. Conclusion The dietary structures of the recruit brigade and the special operations brigade are inappropriate,the intakes of some food and nutrients do not meet the requirements of the current military standards,and the energy supply ratio of the three major nutrients in the special operations brigade is unbalanced. Some of the recruits are either overweight or underweight.
  • Original articles
    Effect of γ-ray radiation on growth of intestinal organoids and biological properties of intestinal stem cells
    LI Xiaoyu, WANG Yongyi, DUAN Min, YE Yumeng, LI Yang, ZUO Hongyan
    2023, 47(9): 650-656. https://doi.org/10.7644/j.issn.1674-9960.2023.09.003
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Objective To investigate the effect of γ-ray radiation on the growth of intestinal organoids and biological properties of Lgr5+ crypt-base columnar cells（Lgr5+ CBCs）, Hopx+ reserve stem cells （Hopx+ rSCs） and Clu+ revival stem cells （Clu+revSCs）. Methods Intestinal crypts of C57BL/6 mice were isolated in vitro and randomly divided into the control group with sham radiation and radiation groups radiated with 5 and 10 Gy γ-ray respectively. Organoids were collected 1 and 6 h after radiation and made into frozen sections, while DNA damage and apoptosis of organoids were detected by immunofluorescence staining of γH2AX and TdT mediated nick end labeling. The growth state of organoids was observed 1, 3 and 5 days after radiation, followed by morphological analysis of the bud ratio and surface area. The cell viability of organoids was examined by CellTiter 3D cell viability assay kit. The expressions of ki67, Apoa4 and Clu in organoids were observed via immunofluorescence staining. The mRNA expressions of APOA4, CLU, LGR5 and HOPX were assessed by real-time fluorescence quantitative PCR （qRT-PCR）. Results Compared with group C, the bud ratio and cell viability of organoids in 5 Gy and 10 Gy groups were significantly reduced one, three and five days after radiation （P<0.001, P<0.05）, and the surface area of organoids was significantly decreased three and five days after radiation （P<0.05）. DNA damage was detected 1 h after radiation and apoptosis was evident 6 h after radiation in 5 Gy and 10 Gy groups （P<0.001）, especially in the 10 Gy group （P<0.001）. Fewer ki67+ proliferation cells but more Apoa4+ mature intestinal epithelial cells and Clu+ stem cells were observed in 5 Gy and 10 Gy groups one,three and five days after radiation,the expressions of APOA4 and HOPX mRNA were significantly higher （P<0.05）, but LGR5 mRNA in organoids was significantly lower than in group C （P<0.05）. CLU mRNA increased significantly one day after radiation （P<0.001）. These changes were more significant in the 10 Gy group than in the 5 Gy group （P<0.05）. Conclusion γ-ray radiation significantly inhibits the growth of intestinal organoids in mice and induces DNA damage and cell apoptosis. Lgr5+CBCs are sensitive to ionizing radiation, while Clu+revSCs and Hopx+rSCs are resistant.
  • Original articles
    Protective effect of 1,2-propanediol against radiation induced oral mucositis
    JIANG Ying, YAN Fulin, WANG Xun, XING Shuang, TU Yanhong, SONG Ruohui, SHEN Xing, YU Zuyin
    2023, 47(9): 657-661. https://doi.org/10.7644/j.issn.1674-9960.2023.09.004
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Objective To assess the protective effect of 1,2-propanediol against radiation-induced oral mucositis in mice. Methods A murine model was established by subjecting the head and neck of mice to a dose of 17 Gy X-rays, and the impact of intraperitoneal prophylaxis with 1,2-propanediol at a dosage of 6 g/kg （administered 3 h prior to irradiation） on radiation-induced oral mucositis in mice was assessed. Sixteen male C57BL/6J mice were randomly and evenly divided into the irradiation and drug administration group. Survival at 30 days and changes in the month after irradiation were recorded. Another 12 mice were divided into the normal group, irradiation group and drug administration group, with 4 mice in each group. These mice were sacrificed 7 days after irradiation. The oral mucosa was stained with toluidine blue, HE staining, Ki67 and P63 immunohistochemistry. Results All the mice in the control group died within 11 days under 17 Gy irradiation. 1,2-propanediol administration significantly improved the survival of mice, and the 30 d survival rate reached 37.5%. The results of morphological observation showed no ulcers on the tongue of mice in the control group, but revealed （0.06±0.01）cm2 ulcers on the tongue of mice in the irradiation group, and the ulcers were （0.01±0.01）cm2 on the tongue of mice in the 1,2-propanediol group at day 7 after irradiation. The histopathological results showed that the structure of tongue mucosa epithelium in the irradiation group was severely damaged with thinner epithelial thickness, but was conserved with less thinned epithelial thickness in the 1,2-propanediol group. Ki-67 and P63 immunohistochemical results showed that keratinocyte proliferation cells and keratinocyte stem cells at day 7 after irradiation in the irradiation group were barely visible, while those in the 1,2-propanediol group showed significant proliferation and regeneration. Conclusion 1,2-propanediol has protective effect on radiation damage in mice with radiation-induced oral mucositis.
  • Original articles
    Structural changes of advanced chromatin in hepatitis B virus after stable transfection
    JIANG Xiuyu, HAO Xiaojuan, ZHOU Zhe, WANG Xuejun, YANG Qian
    2023, 47(9): 662-667. https://doi.org/10.7644/j.issn.1674-9960.2023.09.005
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Objective To investigate the three-dimensional genomic spatial structural alterations in hepatocellular carcinoma cells which are stably transfected with hepatitis B virus （HBV） genome. Methods Bridge Linker Hi-C （BL-Hi-C） was used to obtain information about genomic 3D spatial interactions in HepG2.CWcells stably transfected with HBV and in control cells HepG2. The differentially expressed genes affected by 3D structural alterations were analyzed via transcriptome sequencing. CCK-8 assay and Kaplan-Meier survival analysis were used to analyze the rate of cell proliferation and the function of genes affected by structural alterations of the 3D genome, respectively. Results Information about genomic 3D interactions was obtained from BL-Hi-C assay of HepG2.CW and HepG2 cells. Translocations between chromosome 16 and chromosome 22 and fusion variants of chromosomal topologically associated domains （TADs） were identified at 50kb resolution in HepG2.CW cells compared to HepG2 cells. Higher proliferation activity was detected in HepG2.CW cells as analyzed by the cell growth curve. The expression levels of chromosome 22 open reading frame 34 （C22ORF34） and myo-inositol oxygenase （MIOX） which were located in the chromosomal translocation and TAD-associated regions were found to be significantly changed between HepG2.CW and HepG2 cells.Survival analysis suggested that C22ORF34 and MIOX were related to expected survival of patients. Conclusion Chromosome translocation and 3D genome structureare significantly changed in cells stably transfected with HBV, which may affect the expressions of C22ORF34 and MIOX.
  • Original articles
    Construction of long noncoding RNA SNHG1 knockout cell line using CRISPR/Cas9 system and its function detection
    WANG Tiantian, TAI Fumin, GE Changhui, ZHENG Xiaofei, FU Hanjiang
    2023, 47(9): 668-673. https://doi.org/10.7644/j.issn.1674-9960.2023.09.006
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    Objective To construct a long noncoding RNA （lncRNA） small nucleolar RNA host gene 1 （SNHG1） knockout A549 cell line using the CRISPR/Cas9-mediated homology-directed repair mechanism and to detect the biological function of SNHG1. Methods Small guide RNA （sgRNA） targeting the SNHG1 gene was designed and cloned into the LentiCRISPR v2 plasmid, named Cas9-sgSNHG1 plasmid. The 5′ homology arm （5′Arm） and 3′ homology arm （3′Arm） sequences were amplified from A549 cell genome DNA while the pEGFP-Blast-polyA sequence was amplified from pEGFP-Blast plasmid. Three sequences were linked as 5′Arm-pEGFP-Blast-polyA-3′Arm by overlap PCR, and the product was cloned into pUC18 plasmid by Gibson Assembly, named SNHG1 homologous recombinant template plasmid. The two recombinant plasmids were co-transfected into A549 cells before cell lines stably inserted into pEGFP-Blast-polyA were screened with Blasticidin （Blast）. Real-time quantitative fluorescence PCR （qPCR） was used to detect the knockout efficiency of SNHG1, while CCK-8 and colony formation assay were used to detect cell proliferation and the ability of clone formation. Results Cas9-sgSNHG1 and SNHG1 homologous recombinant template plasmids were constructed, and A549 cell lines with high knockout efficiency were screened. SNHG1 knockout inhibited the proliferation and clone formation of A549 cells. Conclusion SNHG1 knockout A549 cell lines have been constructed, and the function of SNHG1 oncogene is significantly inhibited. CRISPR/Cas9-mediated homology-directed repair can effectively inhibit the expression of long noncoding RNA, which is an important tool for studying long noncoding RNA function.
  • Original articles
    Effects of hypoxia on eNOS activity of endothelial cells and proliferation and migration of smooth muscle cells in a co-culture system
    CAO Yan, HE Yunling, LI Jian, CHEN Ying, WANG Yufei, WU Liying
    2023, 47(9): 674-679. https://doi.org/10.7644/j.issn.1674-9960.2023.09.007
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    Objective To investigate the effects of hypoxia on eNOS activity in endothelial cells and on the proliferation and migration of smooth muscle cells in a co-culture model. Methods A co-culture model of human pulmonary artery endothelial cells （HPAEC） and human pulmonary artery smooth muscle cells （HPASMC） was established. After exposure to hypoxia （3%O2） or normoxia （20%O2）, the effects of hypoxia on proliferation and migration of HPASMC were examined by CCK-8 assay, Ki-67 and EdU staining, Western blotting and Transwell. The effects of hypoxia on NO release and eNOS activity were detected by the testing kit and Western blotting respectively. The role of NO in regulating the proliferation and migration of HPASMC was further verified using sodium nitroprusside （SNP） which could generate NO in the medium. Results Hypoxia could significantly promote the proliferation and migration of HPASMC in the co-culture model while inhibiting the eNOS activity and NO production in HPAEC. However, supplementation of SNP in HPAECs could significantly reverse the proliferation and migration of HPASMC caused by hypoxia. Conclusion Hypoxia reduces the production and secretion of NO by inhibiting the activity of eNOS in HPAECs in the co-culture system, and the reduction of NO ultimately promotes the proliferation and migration of HPASMC.
  • Original articles
    Roles of deoxycholic acid in promoting intestinal Th17 infiltration and inducing intestinal inflammation in mice
    LI Dan, TIAN Chunyan, WU Jin
    2023, 47(9): 680-685. https://doi.org/10.7644/j.issn.1674-9960.2023.09.008
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    Objective To explore the effect of high level deoxycholic acid （DCA） on the infiltration of intestinal T helper cell 17 （Th17） in mice and on the induction of intestinal inflammation. Methods Fourteen wild-type C57BL/6 mice were randomly divided into two groups： the normal diet group and 0.2% DCA-supplemented diet group, with 7 mice per group. After 6 weeks, colon tissue sections were stained with hematoxylin-eosin （HE） and evaluated pathologically. Flow cytometry was used to analyze the proportion of Th17 in mesenteric lymph nodes, immunofluorescence staining was adopted to detect Th17 infiltration in colon tissue, and real-time quantitative PCR （qPCR） was performed to measure the mRNA expressions of interleukin-17（IL-17）, including IL-17A and IL-17F. Results Six weeks of 0.2% DCA-supplemented diet feeding could increase intestinal inflammatory response in mice （P<0.01）, and the proportion of Th17 in mesenteric lymph nodes was significantly increased compared to the normal diet group （P<0.05）. Th17 infiltration in colon increased significantly upon DCA-feeding （P<0.001）, and the expression levels of pro-inflammatory cytokine IL-17A and IL-17F were significant elevated （P<0.05）. Conclusion High level DCA can promote the infiltration of Th17 in intestines and contribute to the induction of intestinal inflammation.
  • Original articles
    Transcription characteristics of human endogenous retrovirus K in leukemia and lymphoma patients
    LI Tianfu, LI Hanping, WANG Xiaolin, QIAN Kun, DOU Liping, LI Jingyun, LI Lin
    2023, 47(9): 686-691. https://doi.org/10.7644/j.issn.1674-9960.2023.09.009
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    Objective To detect and analyze the transcription level of human endogenous retrovirus K （HERV-K） in peripheral blood mononuclear cells （PBMCs） of leukemia and lymphoma patients, identify the integration sites expressed in patients, analyze the expression level of each site, and screen out the HERV-K integration sites with significantly-changed expression levels in patients compared with healthy controls. Methods PBMC samples from leukemia and lymphoma patients were used as the experimental group, while those of healthy controls served as the control group. The transcription level of HERV-K in PBMCs was detected via real-time PCR, and the HERV-K integration sites were identified via PCR amplification and next generation sequencing. The differences in expressions at HERV-K integration sites between the two groups of patients were analyzed. Results The transcription levels of HERV-K gag, pol and env in PBMCs of leukemia and lymphoma patients were significantly higher than those of healthy controls （P<0.0001）. There were respectively 86, 81 and 77 integration sites of HERV-K gag, pol and env in 23 pairs of chromosomes from patients with leukemia or lymphoma and healthy controls. Compared with healthy controls,20 sites were up-regulated and 3 sites were down-regulated in leukemia patients, compared with 12 and 8 sites for lymphoma patients. Biological functions varied according to integration sites. Conclusion The transcription level of HERV-K genes in PBMCs of leukemia and lymphoma patients is significantly increased, and the integration sites of HERV-K with alterations in patients are identified. These findings are expected to help explore the role of HERV-K in the development of leukemia and lymphoma and find new treatment strategies with of HERV-K as the target.
  • Original articles
    Establishment of a novel air-liquid model based on the co-culture of A549 cells and endothelial cells
    ZHANG Xuemei, YANG Hui, ZENG Quan, XI Jiafei, YUE Wen, YAN Xinlong, ZHOU Junnian
    2023, 47(9): 692-699. https://doi.org/10.7644/j.issn.1674-9960.2023.09.010
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    Objective To investigate the effects of air-liquid interface（ALI）co-culture of human type Ⅱ alveolar epithelial cell A549 and endothelial cells on the phenotype and function of A549 cells in the presence of endothelial cells. Methods An air-liquid co-culture model was established using the transwell culture system, in which A549 cells were cultured on the upper surface of transwell membrane and endothelial cells on the bottom surface of transwell membrane. Real-time quantitative PCR（RT-qPCR） was performed to detect the expression levels of surfactant associated proteins, intercellular junctions and proteins associated with SARS-CoV-2 invasion in A549 cells after ALI co-culture with or without endothelial cells,while flow cytometry was performed to detect the infection rate of recombinant lentivirus particles encoding the truncated S protein in the cells. Results Comparative analysis suggested that the EA.hy926 endothelial cell line was preferable for establishing a new ALI co-culture model than primary endothelial cells HUVEC. The traditional ALI culture up-regulated the expressions of surfactant protein SP-B, SP-D,E-cadherin and tight junction protein 1 （ZO-1）, and SARS-CoV-2 receptorsneuropilin 1（NRP1）,asialoglycoprotein receptor 1（ASGR1） and kringle containing transmembrane protein 1（Kremen1）,while ALI co-culture with endothelial cells significantly up-regulated these genes （P<0.05）. The cell infection rate of model virus particles increased significantly compared with traditional ALI culture in A549 cells （P<0.05）. Conclusion An air-liquid co-culture model of A549 cells and EA.hy926 cells has been established,which provides a new pathological model for the investigation of lung diseases and drug screening.
  • Original articles
    Knowledge maps of BMJ Military Health
    YOU Weijie, GUO Qing, WU Jianhu, LIU Dongyan, YU Chaoqi, ZHANG Xiangyang
    2023, 47(9): 700-705. https://doi.org/10.7644/j.issn.1674-9960.2023.09.011
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    Objective To analyze the status quo and developments of British BMJ Military Health since 2011 using knowledge maps and other tools so as to help related health care institutions in China to keep track of British military medicine. Methods The Web of Science Core Collection was searched for articles from BMJ Military Health published between 2011 and 2022,and a total of 1475 pieces of literature were retrieved. Such knowledge map analysis tools as Hiscite,VOSviewer and CiteSpace were used for the research. Results The cited frequency of BMJ Military Health journals continued to trend up year by year,indicating that British military medicine received more attention. Royal Defense Medical Center,Birmingham University,University of London,Imperial College London and King′s College London were the leading contributors. The priorities of research involved the treatment of war wounds,maintenance of mental health,and the prevention and control of infectious diseases. Conclusion Constructing the knowledge maps of BMJ Military Health can help researchers gain insights into the developments of the journal.
    • Reviews
  • Reviews
    Research progress in exosomes derived from different sources and used in wound healing
    MENG Tianjiao, DU Li, ZHAO Xirong
    2023, 47(9): 706-709. https://doi.org/10.7644/j.issn.1674-9960.2023.09.012
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    Wound repair remains a clinical challenge to skin regeneration. Exosomes, as nano-sized extracellular vesicles secreted by a variety of cells, retain complete cellular characteristics. Current studies have found that exosomes secreted by a wide range of cells have different biological components and functional characteristics, and play important roles in various pathophysiological processes. Exosomes can help accelerate wound repair by inhibiting inflammation, facilitate cell proliferation and migration, promote angiogenesis, and inhibit scar formation. This article reviews the research progress in exosomes from different sources that are used in wound repair.
  • Reviews
    Research progress on the role of MHC-Ⅱ in viral infection
    ZHOU Jie, LÜ Zhonglin, WEI Yinxiang, HAN Gencheng
    2023, 47(9): 710-713. https://doi.org/10.7644/j.issn.1674-9960.2023.09.013
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    Viral infectious diseases seriously endanger people′s life and health. The outbreak of coronavirus disease 2019（COVID-19） infection in recent years has brought serious harm to people′s life, health, work and life. Major histocompatibility complex Ⅱ（MHC-Ⅱ） molecules are mainly expressed on professional antigen presenting cells such as B cells, mononuclear macrophages and dendritic cells, and participate in the regulation of humoral and cellular immune responses. More and more studies have shown that MHCⅡ plays an important physiological and pathological role in antiviral infection and viral immune escape, but its mechanism is still unclear. This article reviews the research progress of MHC-Ⅱ in viral infection.
  • Reviews
    Research progress in mesenchymal stem cells in the treatment of lung injury
    MAO Zhuang, WANG Hua
    2023, 47(9): 714-720. https://doi.org/10.7644/j.issn.1674-9960.2023.09.014
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    The increasing incidence and mortality of respiratory diseases have made them a global public health issue. With the development of regenerative medicine,cell therapy has become a new strategy to treat lung diseases. Mesenchymal stem cells are pluripotent adult stem cells with strong ability of self-renewal and differentiation. Due to their biological characteristics,low immunogenicity and abundant sources,mesenchymal stem cells are ideal seed cells for treating acute and chronic lung diseases. This article reviews the possible mechanisms for and current research on mesenchymal stem cells in the treatment of acute and chronic lung injury such as COVID-19,acute lung injury/acute respiratory distress syndrome,chronic obstructive pulmonary disease and asthma.
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