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  • Expert review
    Research concepts and medical support strategies of battlefield acoustics based on military operational medicine
    CUI Bo, ZHU Ying-wen, SHE Xiao-jun, GAO Xiu-jie, MA Ke-feng, WANG Kun, FU Bo, ZHENG Peng-fang, YANG Hong-lian, WANG Xiao-ming, LIU Hong-tao, LI Zhe
    2022, 46(10): 721-725. https://doi.org/10.7644/j.issn.1674-9960.2022.10.001
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Accelerated integration of mechanization, informatization and intelligentization will undoubtedly usher in new concepts on military medical research. The acoustic environment, as a typical physical factor of the military artificial environment, has the properties of an energy and information carrier, and underlies informatization-related operations. Research on battlefield acoustics within military operational medicine has to take into consideration the role of acoustic properties and operation efficiency, and learn from innovative ideas and technologies related to physical acoustics,physiological acoustics, psychoacoustics and acoustic ecology, build a research system for control and application of acoustical environments, and ensure the health of operators and better performance of informatization of man-machine systems.
    • Original articles
  • Original articles
    Protective effect of apoptosis antagonist AS1501 against acute radiation sickness mice
    CAO Lin-chao, ZHAO Dan-yang, ZHENG Ying, GAN Hui, MENG Zhi-yun, DOU Gui-fang, GU Ruo-lan
    2022, 46(10): 726-731. https://doi.org/10.7644/j.issn.1674-9960.2022.10.002
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    Objective To study the role of death receptor 5（DR5） - mediated apoptosis in radiation injury and the feasibility of DR5 being used as a potential target for the development of new anti-radiation drugs. Methods Forty-five C57BL/6 mice were randomly divided into the model control group, normal control group, positive control group, AS1501 low （5 mg/kg）, medium （10 mg/kg） and high dose groups （15 mg/kg） respectively. The 3-month survival rate of different groups after exposure to 8.5 Gy 60Co-γ ray was observed. After mice were treated with 4.0 Gy 60Co-γ ray single whole-body irradiation, AS1501 15 mg/kg was injected into the caudal vein at 2 h, 1 d, 4 d, 7 d, 11 d and 14 d post-irradiation. The number of bone marrow nucleated cells （BMNCs） was measured by cell count and the organ indexes of the spleen and thymus were calculated. The number of bone marrow hematopoietic stem/progenitor cells （LSK/LK） was measured by flow cytometry at 2 h, 1 d and 4 d post-irradiation. The immunofluorescence double labeling was used to detect the changes in expressions of DR5 protein and the apoptosis levels in the thymus at 3 d post-irradiation. Results AS1501 could effectively increase the 3-month survival rate of ARS mice after high-dose 60Co-γ ray irradiation,in a dose-dependent manner. The administration of AS1501 15 mg/kg could significantly increase the organ indexes of the thymus and promote BMNC in ARS mice at 1-11 d post-irradiation （P<0.05）. The LSK/LK level was also significantly increased at 2 h post-irradiation （P<0.05）. Immunofluorescence double labeling test showed that the expression of DR5 protein and the level of apoptosis in thymus tissue of ARS mice were obviously up-regulated at 3 d post-irradiation, and that AS1501 could inhibit the level of apoptosis in thymus tissue of the ARS mice. Conclusion DR5 apoptosis antagonist AS1501 can effectively reduce the early injury to the hematopoietic immune system in ARS mice, promote the recovery of immune organs, and improve the survival rate of ARS mice of the severe bone marrow type, suggesting that TRAIL-DR5 mediated apoptosis probably plays an important role in the biological damage effect of acute radiation injury.
  • Original articles
    Molecular mechanism of abnormal expression of IL-1β in skeletal muscle induced by sleep deprivation
    ZHANG Min, MENG Fan-fei, CHEN Yuan-lian, HUANG Xin, XING Chen, LIU Kun, SONG Lun
    2022, 46(10): 732-736. https://doi.org/10.7644/j.issn.1674-9960.2022.10.003
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    Objective To explore the effect of sleep deprivation （SD） on expressions of inflammatory factors in skeletal muscle and related mechanisms. Methods Male rats weighing about 180-200 g were randomly divided into the control group and experimental group（subjected to SD for 72 hours）. Rat skeletal muscle tissues of each group were collected before the expression levels of inflammatory molecules （IL-1β,IL-15,TNF­α） and related signaling proteins （XBP1s,IRE1α） were detected by RT-PCR and Western blotting assay. Moreover, C2C12 myotube cells were transfected with siRNA targeting the inflammatory response-related proteins and the same detection was performed. Results After SD, the expression level of interleukin-1β （IL-1β） in skeletal muscle was increased along with the upregulation of endoplasmic reticulum stress-related proteins XBP1s and IRE1α. Transfection of XBP1s or IRE1α siRNA into C2C12 myotubes significantly inhibited the transcriptional activation of IL-1β. Conclusion SD induces IRE1α/XBP1s pathway activation in skeletal muscle to mediate the increase in the expression of inflammatory factor IL-1β.
  • Original articles
    Chronic stress-mediated Plexin A1-JAK2-STAT3 pathway stimulates VEGF secretion and promotes gastric cancer angiogenesis
    LIU Ying, CHENG Die, PENG Yu-qiao, PANG Jia-yu, LU Yan-jie, LI Yang, LI Yu-hong
    2022, 46(10): 737-743. https://doi.org/10.7644/j.issn.1674-9960.2022.10.004
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    Objective To explore the effects of isoproterenol （ISO）-simulated chronic stress on gastric cancer angiogenesis through the Plexin A1-JAK2-STAT3 pathway. Methods The lentivirus or pathway inhibitor was used to intervene in the expression of Plexin A1 or JAK2-STAT3 signal pathway in MGC-803 gastric cancer cells respectively before ISO at the concentration of 20 μmol/L was also used to treat gastric cancer cells for 12 hours. The gastric cancer cell culture medium was collected, and the level of vascular endothelial growth factor （VEGF） secreted by gastric cancer cells was detected by radioimmunoassay. The gastric cancer cell culture medium was collected a second time to continue to culture vascular endothelial cells before the proliferation, migration and tube forming ability of vascular endothelial cells EA.hy926 were detected using the CCK-8 method, Transwell method and angiogenesis experiment respectively. Western blotting was used to detect the protein expressions of Plexin A1 and VEGFR2 in vascular endothelial cells. Results After lentivirus interfered with Plexin A1, the secretion level of VEGF in gastric cancer cells was significantly reduced. After vascular endothelial cells were cultured with gastric cancer cell culture medium that interfered with the expression of Plexin A1, the expressions of VEGFR2 and Plexin A1 in vascular endothelial cells were significantly reduced, so were the proliferation, migration, and tube-forming abilities of these cells. Inhibition of JAK2-STAT3 in gastric cancer cells signaling pathway resulted in significant reduction of the secretion level of VEGF in gastric cancer cells and of the proliferation, migration and tube-forming abilities of vascular endothelial cells cultured with gastric cancer cell culture medium. Conclusion Chronic stress promotes the angiogenesis of gastric cancer by stimulating the secretion of VEGF through the Plexin A1-JAK2-STAT3 signaling pathway in gastric cancer cells.
  • Original articles
    Purification and characterization of fluorescently-labelled Clostridium perfringens ε toxin fusion protein
    GUO Zi-shuo, LÜ Ruo-mei, HUANG Jing, LIU Ting-ting, CHEN Ming, KANG Lin, WANG Jing-lin, XIN Wen-wen
    2022, 46(10): 744-748. https://doi.org/10.7644/j.issn.1674-9960.2022.10.005
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    Objective To investigate the properties of Clostridium perfringens epsilon toxin （ETX） by expressing and purifying ETX with eGFP or mScarlet tag. Methods ETX with eGFP or mScarlet tag prokaryotic expression plasmids were constructed, transformed in E.coli, and finally purified using a Ni2+chelating affinity chromatography resin column. The toxicity of ETX with different tags was determined using the MTS method. A confocal microscope was used to observe the binding of mScarlet-ETX to HaCaT cells, HEKa cells and red blood cells of different species. Results ETX with eGFP or mScarlet tag was expressed, and high purity of eGFP-ETX and mScarlet-ETX was obtained through purification. eGFP-ETX was confirmed to be less toxic than His-ETX by cytotoxicity assay, and mScarlet-ETX toxin was almost equally toxic. The optimal fluorescence values of eGFP-ETX and mScarlet-ETX were also determined. After mScarlet-ETX was applied to HaCaT cells and HEKa cells,it was observed that mScarlet-ETX bound to the HaCaT cells and HEKa cells, confirming that HaCaT cells and HEKa cells were sensitive to ETX. The mScarlet-ETX was also applied to red blood cells of humans, mice and rabbits, and the binding of mScarlet-ETX to human red blood cells was observed. Conclusion mScarlet-ETX has good toxin activity and can be used in ETX toxicity studies.
  • Original articles
    The role of polymorphonuclear neutrophils in the process of acute lung injury in mice infected via aerosolized intratracheal inoculation of Pseudomonas aeruginosa
    XIAO Nan, ZHANG Zhi-peng, LI Sha, DENG Meng-yun, WANG Yi-feng, ZONG Fu-liang, SU Duo, ZHOU Dong-sheng, YANG Hui-ying
    2022, 46(10): 749-757. https://doi.org/10.7644/j.issn.1674-9960.2022.10.006
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    Objective To explore the role of polymorphonuclear neutrophils （PMN） in acute lung injury （ALI）in mice infected by aerosolized intratraheal inoculation of Pseudomonas aeruginosa（PA）. Methods A mouse model of depleted PMN was constructed via intraperitoneal injection of anti-Gr-1 antibody, while PMN depletion efficiency was assessed by flow-cytometry and immunofluorescence. The survival of mice was observed after infection, the pathological degree of lungs was evaluated, and the changes in bacterial loads of lungs and cytokine concentration in alveolar lavage fluid were detected to analyze the role of PMN in the process of ALI. Results Flow cytometry and immuno-fluorescence results showed that a mouse model of depleted PMN was constructed. Compared with control mice, the mortality and lung bacterial loads of PMN-depleted mice were increased after infection. There was only a small amount of inflammatory cell infiltration in the lung but a large number of bacteria were visible in the alveolar lumen. The expressions of pro-inflammatory factors such as interleukins-6（IL-6）, IL-1β, IL-18, IL-17A, IL-22 and tumor necrosis factor-α （TNF-α） were upregulated, so were the expressions of chemokines such as monocyte chemotactic protein-1 （MCP-1）, macrophage inflammatory protein-2 （MIP-2） and granulocyte-macrophage colony-stimulating factor （GM-CSF）. The expressions of anti-inflammatory factors such as IL-4, IL-10 and IL-13 were also upregulated in alveolar lavage fluid. Conclusion The recruitment of PMN effectively inhibits the colonization of bacteria during infection. The compensatory elevation of multiple cytokines in mice after PMN depletion suggests that PMN can inhibit the excessive development of inflammation during ALI and PMN can be an important regulator of inflammatory and immune responses.
  • Original articles
    Regulation of expressions of interferon-β by Yersinia pestis YopT mutants
    YAN Qiu-lin, LI Jing-fei, ZHANG Xiao-juan, DONG Ying, CHANG Gui-juan, LI Tong-ju, WEI Cong-wen
    2022, 46(10): 758-762. https://doi.org/10.7644/j.issn.1674-9960.2022.10.007
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    Objective To construct YopT mutants including F296A,G298A,H303A,Y304F,F296A-G298A and H303A-Y304F and to detect their regulation of expressions of interferon-β（IFN-β） was. Methods Using pCMV-Myc-YopT as the template,the fragments of six mutants of yopT were amplified by PCR before being digested by endonuclease and ligated into the pCMV-Myc vector to form recombinant plasmids that were transfected into HEK293T cells respectively,and the expression level of YopT mutants was detected by immunoblotting and the effect of wild type YopT and its six mutants on the expression of IFN-β was detected by luciferase reporter gene assay. Results Six yopT mutated recombinant plasmids were successfully expressed in HEK293T cells. It was found that YopT-F296A,G298A,H303A,Y304F,F296A-G298A and H303A-Y304F could inhibit the expression of IFN-β reporter gene. YopT-C139R had no significant inhibitory effect on the expression of IFN-β. Conclusion YopT 139 is the key site affecting the expression of IFN-β,while the sites of 296,298,303,304 are not the key site affecting the expression of IFN-β,and its virulence function is of vital importance in studies on the innate immune evasion strategy adopted by Yersinia pestis.
  • Original articles
    Construction of a stable transgenic cell strain for packaging of West Nile pseudovirus
    LU Xing, SUN Ying, CHEN Guo-jiang, WANG Jing, QIAO Chun-xia, LUO Long-long, LI Xin-ying, LIU Cheng-hua, SHEN Bei-fen, FENG Jian-nan, XIAO He
    2022, 46(10): 763-767. https://doi.org/10.7644/j.issn.1674-9960.2022.10.008
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    Objective To establish a stable cell strain that can package West Nile pseudovirus in order to facilitate the preparation of West Nile pseudovirus. Methods BHK21 cells were transfected with recombinant plasmid pcDNA3.1-CME expressing the structural proteins C, prM and E of West Nile virus （WNV）before endergoing pressured screening with G418 （1 mg/ml）. The three structural proteins C, prM and E expressed in stable cell strains was identified by flow cytometry, indirect immunofluorescence staining （IFA） and Western blotting （WB）. The stable cell strain were infected with the 103-diluted WNV pseudovirus supernatant containing the luciferase reporter gene, and the infected cells were lysed to detect the luciferase activity. Results and Conclusion The stable cell strain expressed three structural proteins： C, prM and E. The supernatant secreted by the WNV pseudovirus was further infected with BHK21 cells that were to produce fluorescence, confirming that the stable cell strain could package the WNV pseudovirus.
  • Original articles
    Adenoid immune cell heterogeneity in children based on single-cell transcriptome sequencing
    LIU Shi-yu, CUI Jing-hua, FU Tong-tong, LIU Yan-fen, YUAN Jing, YANG Rui-fu
    2022, 46(10): 768-775. https://doi.org/10.7644/j.issn.1674-9960.2022.10.009
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    Objective To chart a single-cell transcriptome atlas of adenoids in children and reveal the heterogeneity of adenoid immune cells. Methods Adenoid tissue was obtained clinically and converted into a cell suspension. Single-cell whole transcriptome data was obtained through the BD RhapsodyTM single-cell system and Illumina next-generation sequencing technology. Finally, R and Python software was used for cluster analysis, pseudotime trajectory analysis, functional analysis and single-cell regulatory network inference and clustering（SCENIC） analysis. Results A total of 14 618 high-quality cells were obtained, which could be clustered into 8 types. After non-negative matrix factorization, 13 and 12 subsets of T and B cells, which were the dominant cell types, were clustered, respectively, and their pseudotime trajectories, regulatory networks of transcription factors and pathway activity were analyzed. Conclusion In this study, a detailed adenoid single-cell transcriptome atlas is drawn, which can shed light on the heterogeneity of pediatric adenoid immune cells.
  • Original articles
    Content determination of dual orexin receptor antagonist WEB by HPLC
    WANG Si-yi, YANG Jian-yun, XIAO Bing-kun, MIAO Xiao-yao, LI Zhi-heng, HUANG Rong-qing
    2022, 46(10): 776-779. https://doi.org/10.7644/j.issn.1674-9960.2022.10.010
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    Objective To establish an HPLC method for the determination of contents of cyclopropane - diorexin receptor antagonists （1R, 2S）-2-（3-chloro phenyl）-N-（4-fluoro phenyl）-2-[（6-methyl pyridine-3-yl）oxymethyl ] cyclopropane-1- formamide （WEB）. Methods The Diamonsil -C18 column （4.6 mm×250 mm, 5 µm）was used. The mobile phase was A（acetonitrile,methanol and ethanol at the ration of 4∶1∶1, V/V/V）-B（20 mmol/L,pH adjusted to 4.2 with KH2PO4） under gradient elution. The flow rate was 1 ml/min, column temperature 40℃,detection wavelength 221 nm and the sample volume was 20 µl. Results In the range of 0.0005-1.0 mg/ml, the peak area showed a good linear relationship with the concentration（r= 0.9997）. The detection limit was 0.2 µg/ml, the limit of quantification was 0.5 µg/ml, and the precision and repeatability of the method met requirements. The average recovery was 101.23%（RSD was 0.71%, n=9）. Conclusion This method can be used for WEB API quality control and stability studies.
  • Original articles
    Distribution and drug resistance of pathogenic microorganisms isolated from patients with acute poisoning
    YIN Xiu-yun, ZENG Li-jun, XU Jian-min, JIANG Qian, YANG Zhe, LI Hao-lian, WANG Wen-cai, LI Bo, CHEN Jian-kui
    2022, 46(10): 780-785. https://doi.org/10.7644/j.issn.1674-9960.2022.10.011
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    Objective To investigate the distribution and drug resistance of pathogenic bacteria isolated from patients with acute poisoning so as to provide reference for the diagnosis,prevention and treatment of acute poisoning infections. Methods A retrospective analysis of pathogenic microorganism infections in patients with acute severe poisoning between 2015 and 2020 was conducted. Results The most common type of acute poisoning was pesticide poisoning, followed by drug poisoning. The infection sites of acute poisoning were mainly the lower respiratory tract （69.8%） and urinary tract （18.2%）. Among the 1129 strains of pathogenic bacteria, bacteria accounted for 78.48%, and fungi 21.52%. The most common pathogens were Acinetobacter baumannii, Candida albicans, Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli. CRAB and MRCNS had higher isolation rates,which were 80.6% and 87.5% respectively. The bacterial susceptibility results showed that the commonly used antibacterial drugs, such as carbapenems, third-generation cephalosporins, fourth-generation cephalosporins, β-lactam complexes and fluoroquinolones, were highly resistant to non-fermenting A.baumannii. The drug resistance rate was above 70%. The resistance rate of K.pneumoniae to carbapenems and β-lactam complexes of piperacillin/tazobactam was low, ranging from 8% to 14%. Among the commonly used antibacterial drugs, the third and fourth generations of cephalosporins and fluoroquinolones were more resistant to E.coli, but more sensitive to K.pneumoniae. Staphylococcus aureus, coagulase-negative Staphylococcus, and Enterococcus faecalis were found to be non-resistant to vancomycin, linezolid and tigecycline. In Enterococcus faecium, the VRE isolation rate was 3.3%. The fungal drug susceptibility results showed that the common Candida spp. were 100% sensitive to both amphotericin B and 5-flucytosine. Fluconazole could be used empirically for Candida species other than C.krusei, which was inherently resistant to fluconazole. Conclusion In this study, it has been found that there is a wide variety of pathogenic bacteria in patients after acute poisoning, and drug resistance is serious and complex. Therefore, in order to increase the success rate of acute poisoning rescue, the diagnosis of related bacterial and fungal infections after poisoning deserves more attention. Based on the bacterial and fungal identification and drug sensitivity test, clinicians can select appropriate antibiotics and other therapeutic drugs to save as many lives as possible.
    • Reviews
  • Reviews
    Animal models for stress-related disorders
    WANG Ting, WU Hai-tao
    2022, 46(10): 786-791. https://doi.org/10.7644/j.issn.1674-9960.2022.10.012
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    Stress-related disorders are a psychiatric disorder of high prevalence and major socioeconomic impact. Despite recent progress, the molecular mechanisms underlying the pathology of stress-related disorders are poorly understood. Animal studies are essential for identifying the mechanisms of stress-related disorders and the research of therapeutic drugs. Although none of the animal models completely resembles human mental disorders, most models mimic many of the features observed in the human situation. In this paper, construction methods, their strengths and weaknesses, symptoms of mental disorders and pathological patterns in animal models of stress-related disorderss are reviewed, which can provide insights into different animal models for the study of stress-related disorders.
  • Reviews
    Recent advances in antioxidant drugs against electromagnetic radiation
    MEN Jun-qi, YAO Bin-wei, GUO Jia-bin, PENG Rui-yun
    2022, 46(10): 792-797. https://doi.org/10.7644/j.issn.1674-9960.2022.10.013
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    In recent years, electromagnetic wave technology has developed rapidly and has been widely used in the military, communication, health care and daily lives. It not only facilitates production and life, but also may harm human health. A large number of studies have shown that electromagnetic radiation can affect or even damage many systems of the body after reaching a high intensity, and ways to protect against such injury have been extensively studied. Oxidative stress is one of the important mechanisms of electromagnetic radiation injury, and also one of the important areas of research on drugs for electromagnetic radiation protection. Antioxidant drugs are the main means of combating oxidative stress. This paper reviews the current research and progress in antioxidant drugs for electromagnetic radiation protection.
  • Reviews
    Progress in multiplex PCR technology for SARS-CoV-2 detection
    LI Qing-chao, FANG Xue-xun, XU Xiao-jie
    2022, 46(10): 798-801. https://doi.org/10.7644/j.issn.1674-9960.2022.10.014
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    SARS-CoV-2 has swept the world. However, it is difficult to detect SARS-CoV-2, because of its variants and co-infection with other viruses.This paper reviews the multiplex PCR methods for detecting SARS-CoV-2. Multiplex PCR is an efficient, accurate, specific and low-cost technology for detection of nucleic acid. It provides a new detection method for SARS-CoV-2 variants and co-infection by combining multiplex PCR with other technologies, such as sequencing and mass spectrometry.
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