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  • Original articles
    In vitro maintenance of hematopoietic progenitor cells derived from human embryonic stem cells via co-culture of AFT024 cells
    ZHU Mei-qi, ZHANG Bo-wen, WU Xu-min, LI Ji-sheng, SONG Fei-ling, FAN Zeng, HE Li-juan, PEI Xue-tao, LI Yan-hua
    2022, 46(8): 561-568. https://doi.org/10.7644/j.issn.1674-9960.2022.08.001
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    Objective To explore the role of AFT024 cells in maintaining the characteristics of human embryonic stem cells （hESCs）-derived hematopoietic progenitor cells in vitro. Methods A culture system was established to induce hESCs to differentiate into hematopoietic progenitor cells. Hematopoietic progenitor cells were generated using a three-step protocol by adding different cytokine in the form of an embryoid body （EB）. hESCs-derived hematopoietic progenitor cells were co-cultured with AFT024 cells, while the control group was cultured without these feeder cells. After four days of co-culture, the number and proportion of hematopoietic progenitor cells and the number of hematopoietic colonies formed were detected by counting the number of total cells and via flow cytometry analysis and colony forming unit （CFU） assay. The expression levels of genes related to proliferation and differentiation of hematopoietic stem/progenitor cells were detected by quantitative real-time polymerase chain reaction（qRT-PCR） to find out whether AFT024 had the ability to maintain the characteristics of hematopoietic progenitor cells in vitro. Results After fourteen days of three-stage induction, hESCs could produce a large number of hematopoietic progenitor cells, and the proportion of CD34+CD43+ cells was about （22.63±2.65）%,compared with about（23.67±2.15）% for CD34+CD45+ cells. Immunofluorescence staining showed that α-SMA and Dlk-1 proteins were expressed in AFT024 cells. The proportion of hematopoietic progenitor cells cocultured with AFT024 cells for four days was significantly larger than that of non-feeder culture group. The results of hematopoietic CFU assay showed that the total number of hematopoietic colonies produced in the co-culture group was significantly higher than that in the control group. Conclusion AFT024 can maintain the characteristic of hESCs-derived hematopoietic progenitor cells in vitro.
  • Original articles
    Down-regulation of autophagy-related protein ATG4B inhibits the intracellular replication of Coxiella burnetii
    ZHANG Jia-ning, FU Meng-jiao, JIAO Jun, OUYANG Xuan, YU Yong-hui, ZHAO Ming-liang, ZHANG Shan, WEN Bo-hai, XIONG Xiao-lu, SUN Yan-song
    2022, 46(8): 569-575. https://doi.org/10.7644/j.issn.1674-9960.2022.08.002
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    Objective To investigate the effects of autophagy-related gene 4B （ATG4B） on the intracellular replication of Coxiella burnetii （C.burnetii） and the development of Coxiella-containing vacuoles （CCVs）. Methods Host cells （THP-1 cells and Vero cells） treated with ATG4B-targeting siRNA served as the experimental group,those treated with non-targeting siRNA served as the negative control group （NC）, and untreated ones served as the mock control. The expression levels of autophagy molecular markers （p62, LC3）in the three groups were detected by Western blotting. The bacterial loads in these groups were detected using qRT-PCR. The development of CCV in these groups was observed using confocal microscopy. Results Compared with the control groups, the autophagy molecular markers （p62, LC3） in the experimental group significantly increased while bacterial loads became much lower, and reduced CCV sizes and a multi-vacuolar CCV phenotype in ATG4B-silenced cells were observed. Conclusion The results suggest that ATG4B play an important role in modulating autophagy, which could be required for the development of CCV and the replication of C.burnetii in host cells. This study can shed light on the relationship between autophagy machinery and the pathogenic mechanism of C.burnetii.
  • Original articles
    Construction and multi-omics analysis of a cell photoaging model
    ZOU Xiao-cang, YU Ren-feng, ZOU Da-yang, WANG Ke-hui, LI Lin-hao, XU Ya-qing, QIN Ri-hui, HE Xiao-ming, LIU Wei, GUO Jin-peng
    2022, 46(8): 576-583. https://doi.org/10.7644/j.issn.1674-9960.2022.08.003
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    Objective To construct a photoaging model of human immortalized keratinocytes（HaCaT） pretreated with serum starvation and explore the biological effects of UVB radiation on the photoaging process of HaCaT cells via multi-omics correlation analysis. Methods HaCaT cells were pretreated using serum starvation method for 24 h before being irradiated with 20 mJ/cm² UVB. Apoptosis was detected by flow cytometry, glutathione and mitochondria membrane potential were detected by high-content screening and cell motility was measured by scratch test to find out whether the photoaging model was properly constructed. HaCaT cells were subjected to high-throughput sequencing before and after light exposure to analyze differentially expressed genes in miRNA genomics and mRNA genomics. Some genes were selected for quantitative real-time PCR（qRT-PCR）, and pathway enrichment of differential genes was performed. Results After UVB irradiation, the number of apoptotic cells was increased, glutathione levels decreased, mitochondrial membrane potential decreased, and the ability of motility and migration was weakened, indicating that the photoaging model was workable under serum-free conditions. A total of 106 intersecting differential genes were obtained through the association analysis of miRNA genomics and mRNA genomics. Kinase suppressor of Ras 1 （KSR1）, dedicator of cytokinesis 1 （DOCK1）, vascular endothelial growth factor A （VEGFA） and myosin light chain kinase （MYLK）-were selected for qRT-PCR verification. The trend before and after UVB irradiation was consistent with the mRNA genomics. KEGG functional enrichment was performed on 106 genes, which were significantly enriched in p53 and focal adhesion signaling pathways. Conclusion After pretreatment with serum starvation, 20 mJ/cm² UVB irradiation can help construct the photoaging model of HaCaT cells. The p53 and focal adhesion signaling pathways are critical to the photoaging process.
  • Original articles
    Changes and roles of astrocyte phagocytic receptor MERTK in stress cognitive dysfunction
    GUO Bo-min, ZHAO Yun, XIE Fang, WANG Xue, SUN Zhao-wei, QIAN Ling-jia
    2022, 46(8): 584-588. https://doi.org/10.7644/j.issn.1674-9960.2022.08.004
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    Objective To observe the changes in astrocyte phagocyte receptors under chronic stress and explore the possible mechanism of cognitive dysfunction induced by chronic stress. Methods Chronic restraint stress （CRS） was used to establish a rat model of cognitive dysfunction induced by stress. The open field test, objective recognition test and water maze experiment were used to observe rats′ behavioral performance. The ELISA method was used for the detection of concentrations of serum stress hormones GC. Western blotting and quantitative real-time polymerase chain reaction（qRT-PCR） were used to detect transcription/translation levels of MERTK and SYP in different areas of the brain. Immunofluorescence was used to detect the expression of SYP in the hippocampus. Results Compared with the Ctrl group, serum and cerebrospinal fluid stress hormone GC levels in the CRS group increased （P<0.05）, while the open field score, objective cognitive index, and score of spatial probe of water maze decreased（P<0.05）. Transcription/translation levels of MERTK in each area of the brain increased（P<0.05）, while synaptic structure markers, and synaptophysin decreased（P<0.05）. Conclusion Chronic stress induces elevated expressions of astrocyte phagocytic receptor MERTK and downregulated expressions of the synaptic structural marker SYP, which may be due to abnormal phagocytic function caused by elevated MERTK.
  • Original articles
    Effect of transcranial direct current stimulation on brain cognitive function under hypoxia
    GUO Da-long, WANG Cong, ZHOU Yu-bin, QIN Yu-fei, YE An-qi, LIU Juan, XIA Yang, YANG Lei
    2022, 46(8): 589-593. https://doi.org/10.7644/j.issn.1674-9960.2022.08.005
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    Objective To investigate the effect of the transcranial direct current stimulation （tDCS） technique on brain cognitive function under a hypoxic environment in order to facilitate the application of tDCS devices. Methods An atmospheric-pressure hypoxic chamber was used to simulate the hypoxic environment of the plateau 3700 meters above sea level. Twenty healthy volunteers were chosen as subjects,who were exposed to a hypoxic environment. Ten of the subjects received tDCS, and the rest underwent sham stimulation. Their response latencies, local cerebral oxygen saturation, the Profile of Mood States, flicker fusion frequency, and altitude sickness scale were measured five times.The differences between them were studied. Results The results showed that tDCS could significantly improve the subjects′ emotional state, especially depression and fatigue. Notably, it could improve vigor for more than 6 hours （P<0.01 from the second to fifth measurements）. In addition, tDCS could improve the alertness of the brain for a short time（P<0.01 from the second and third measurements）. Cerebral oxygen saturation was greatly improved within 4 hours, thus alleviating fatigue （P<0.01 during the second, third and fourth measurements）. Conclusion In a hypoxic environment, tDCS can improve alertness, increase local cerebral oxygen saturation, reduce depression and fatigue and boost energy.
  • Original articles
    Identification of functional amino acids of HIV-1 gp41 CHR peptide CP621-652 and interaction with NHR peptide T21
    ZHAO Zhi-ming, WANG Xiao-hua, YU Shuo, ZHONG Hui-hui, HE Yu-xian, PENG Sheng-ming, DAI Qiu-yun
    2022, 46(8): 594-600. https://doi.org/10.7644/j.issn.1674-9960.2022.08.006
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    Objective To determine the functional amino acids of the first seven amino acids of HIV-1 gp41 CHR peptide CP621-652 at N-terminus and the orientation of complex of CP621-652 and NHR peptide T21, and provide a clue for further modifications of novel HIV-1 gp41 fusion inhibitors. Methods Seven CP621-652 variants containing an alanine （Ala）substitution one by one for the first seven amine acids QIWNNMT were synthesized and their inhibitory activities against HIV-1 were determined. The interactions of CP621-652 variants with gp41 NHR-peptide T21 were determined by circular dichroism spectroscopy, electrophoresis and gel exclusion. The variants of CP621-652 and T21 modified by cysteine at N- or C-terminus and their dimers of disulfide bond were synthesized, the binding orientation of the complex of CP621-652 and T21 was analyzed by oxidative folding and equilibrium strategy. Results The replacement of Q1, W3, M6 and T7 by Ala resulted in significant decrease in fusion inhibitory activity. The results of circular dichroism spectroscopy, electrophoresis and gel exclusion chromatography showed that CP621-652 and its mutant interacted with T21 to form a helical complex. CP621-652 with cysteine at N-terminal and T21 with cysteine at C-terminal was folded to form a high ratio of heterodimers. The exchange equilibrium experiment showed that the heterodimers were more stable than the homodimers. Conclusion Q1, W3, M6 and T7 are the functional amino acids of CP621-652. CP621-652 and T21 form antiparallel helical structures.
  • Original articles
    The mechanism of melatonin in alleviating ulcerative colitis in a mouse model
    ZHANG Su-yue, DU Xin-jian, YANG Le, YIN Ji-ye, LIAO Sha, ZHOU Zhe
    2022, 46(8): 601-605. https://doi.org/10.7644/j.issn.1674-9960.2022.08.007
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    Objective To explore the mechanism by which melatonin alleviates ulcerative colitis （UC） in a mouse model. Methods Thirty male mice （22-25 g, C57BL/6） were randomly divided into the negative control, positive control and drug treatment groups. The weight, faeces and mental status of the mice were recorded. Enzyme-linked immunosorbent assay （ELISA） and fluorescent quantitative polymerase chain reaction （qPCR） were used to detect the level of interleukin 1β （IL-1β） and the expression of inflammatory apoptosis-related genes in colon tissues. Bacterial lipopolysaccharide （LPS） and adenosine triphosphate （ATP） were used to induce macrophage inflammatory apoptosis. The proportion of inflammatory apoptotic macrophages was detected by flow cytometry. Results After melatonin treatment, the proportion of inflammatory apoptotic macrophages decreased from 36.5% to 19.8%, and the IL-1β concentration decreased from 65.5 pg/ml to 40.0 pg/ml compared to the model group. Melatonin-treated mice increased the survival rate of UC to 50%. Melatonin reduced the level of IL-1β from 7.7 pg/mg to 3.46 pg/mg and mRNA expression levels of inflammatory apoptosis-related genes in the colon tissue of UC mice. Conclusion Melatonin inhibits the inflammatory apoptosis of macrophages and exerts an anti-inflammatory effect in a mouse model of UC.
  • Original articles
    MRI analysis of intracranial infiltration of leukemia
    ZHANG Hong-tao, CAI Jian-ming, ZHANG Zhao-xue, TANG Dai-di, GAO Shen, ZHOU Juan, TIAN Ning, SHENG Fu-geng
    2022, 46(8): 606-609. https://doi.org/10.7644/j.issn.1674-9960.2022.08.008
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    Objective To analyze the magnetic resonance imaging （MRI） findings of intracranial infiltration of leukemia in order to advance our understanding and diagnosis of intracranial infiltration of leukemia. Methods The clinical data and MRI findings on 22 patients with leukemia with intracranial infiltration diagnosed in our hospital between February 2009 and December 2019 were analyzed retrospectively. The locations of lesions, extent of involvement, signal characteristics and enhancement features of T2WI, T1WI and DWI were compared and analyzed. Results Intracranial infiltration of leukemia manifested itself as meningeal infiltration and brain parenchyma infiltration on MRI. The manifestations of meningeal infiltration in 13 cases of leukemia fell into four types： the diffuse dural thickening type （n=4）, diffuse dural thickening with local nodule type （n=5）, dural nodular type （n=2） and leptomeningeal thickening type （n=2）. There were 9 cases of cerebral parenchyma infiltration of leukemia, which were divided into two types： the multiple nodular type （n=4） and single nodular type （n=5）. Most of the brain parenchyma infiltrating nodules showed iso-and slightly high-signals on T2WI, iso-and low-signals on T1WI, high-signals on DWI, moderate to obvious enhancement on contrast-enhanced scan, and different degrees of brain edema around the focus. Conclusion MRI findings of intracranial infiltration of leukemia have some characteristics, which can be divided into meningeal and cerebral parenchyma infiltration. MRI is of great value in the detection and diagnosis of lesions.
  • Original articles
    Clinical and pathological analysis of anti-NXP2 antibody-positive idiopathic inflammatory myopathy： a report of 6 cases and review of literature
    CHEN Juan, LIN Ying, LIU Meng-yang, SHI Qiang
    2022, 46(8): 610-615. https://doi.org/10.7644/j.issn.1674-9960.2022.08.009
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    Objective To investigate the clinical manifestations and pathological features of skeletal muscle in patients with idiopathic inflammatory myopathy （IIM） with positive anti-nuclear matrix protein 2（NXP2） antibodies. Methods The clinical data of 6 IIM patients with positive anti-NXP2 antibodies was collected and analyzed retrospectively. Results Myalgia, muscle weakness of neck extensors and proximal extremities were found in all the 6 cases,while dysphagia and respiratory muscle weakness were more common. There were 5 cases with skin changes, including calcinosis cutis and Heliotrope signs on cheeks and in anterior neck areas. The important extramuscular target organs were the hollow organs （the esophagus, stomach, intestinal system and kidney, ureter, bladder system）, which could cause gastrointestinal bleeding and perforation in severe cases. The pathological changes in skeletal muscle were slight, with necrosis of several muscle fibers, atrophy of muscle fibers around the bundle, micro-infarction of muscle fibers, infiltration of inflammatory cells around blood vessels in the muscle interstitial, but infiltration of inflammatory cells in the endomysium was rare. The six patients were treated with hormone combined with immunosuppressive therapy, and muscle strength improved. Case 3 died due to gastrointestinal complications. Conclusion The incidence of IIM with positive anti-NXP2 antibodies is low. Cervical and proximal muscles are easily affected, while the involvement of bulbar and respiratory muscles is relatively common. Gastrointestinal and urinary complications are associated with mortality. The pathological changes in skeletal muscle are relatively slight. High dose hormone shock combined with immunosuppressive therapy is effective.
  • Original articles
    Development of unmanned medical service equipment of U.S. Armed Forces in field：a progress and prospective
    ZHANG Jin-hui
    2022, 46(8): 616-620. https://doi.org/10.7644/j.issn.1674-9960.2022.08.010
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    Using robot technology, especially autonomous and semi-autonomous systems, to develop field unmanned medical equipment is an important means to improve the medical ability to rescue the wounded on the battlefield and reduce the risk of first responders. Since the end of the last century, the U.S. Armed Forces have established a series of projects to develop, integrate or transform robots and unmanned ground / air systems to evacuate battlefield casualties from hostile environments and fires. This paper aims to provide reference for the development of field unmanned medical service equipment of China′s army by analyzing objectives, status quo, existing problems and developments of field unmanned medical service equipment of the U.S. Armed Forces.
    • Reviews
  • Reviews
    Regulation of functions of the immune system by circadian rhythms: research progress
    ZHAO Guo-jie, CUI Bo, SHE Xiao-jun, GAO Xiu-jie, MA Ke-feng, YANG Hong-lian, WANG Ying
    2022, 46(8): 621-626. https://doi.org/10.7644/j.issn.1674-9960.2022.08.011
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    Circadian rhythms are various oscillatory patterns of behavioral and physiological functions with a cycle of roughly 24 h and widely exist at the whole, systemic, organic, cellular and molecular levels from single-cell organisms to higher organisms. In the immune system, significant circadian rhythms are found to exist in many immune functions （migration, chemotaxis, phagocytosis, and immune factor release） and multiple immune parameters （the absolute and relative number of circulating leukocytes and their subpopulations, the number of lymphocytes, cytokinine levels, and humoral immune response, etc.）. The maintenance of circadian rhythms is regulated by the intracellular biological clock, while the biological clock regulates circadian rhythm changes in the immune system via clock genes （clock, bmal1, per, cry） and clock controlled genes （rev-erbα, rorα, dbp, hlf, tef, etc.）, which plays an important role in maintaining immune homeostasis. Studies have shown that some immune related diseases are closely related to abnormal biological clock systems. Therefore, this review summarizes the relationships between the biological clock and the immune system as well as its role in immune-related diseases.
  • Reviews
    The related mechanism of Six1 in tumorigenesis and development
    HUANG Meng-gen-tu-ya, LIN Jing
    2022, 46(8): 627-631. https://doi.org/10.7644/j.issn.1674-9960.2022.08.012
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    As the most extensively investigated member of sine oculis homeobox homolog （Six）transcription factor family, Six1 is involved in the development of many tissues and organs,as well as in tumorigenesis. By binding to specific DNA sites, it specifically regulates the expression of related target genes, and plays a key role in the occurrence, development and prognosis of tumors, which has become the focus of tumor research. Recent studies have shown that Six1 affects the occurrence, development and prognosis of tumors mainly through the following mechanisms. Six1 promotes the proliferation of tumor cells by regulating cell cycle, apoptosis, senescence, stemness and drug resistance. Six1 facilitates the invasion and metastasis of tumor cells by inducing epithelial-mesenchymal transformation of tumor cells, expressions of invasive proteins and angiogenesis of blood vessels and lymphatic vessels. Six1 affects the metabolism of tumor cells by promoting Warburg effect of tumors. This article reviews the related mechanisms by which Six1 promotes the occurrence and development of tumors
  • Reviews
    Research progress in pathogenesis and prevention of age-related hearing loss
    TANG Xiao-xu, DING Zhong-jia, LI Jia-lin, SUN Bao-chun
    2022, 46(8): 632-637. https://doi.org/10.7644/j.issn.1674-9960.2022.08.013
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    Age-related hearing loss （ARHL）, also known as presbycusis,is caused by interactions between a variety of pathogenic factors and primarily associated with genetic factors, oxidative stress, mitochondrial damage, inflammatory reaction and cell apoptosis. In addition to the traditional methods of prevention, auditory assistance devices including hearing aids,cochlear implants and middle ear implants have been widely used in clinic in the recent five years. Gene and stem cell therapy in treating ARHL also have made new progress. So far, many advances have been made related to pathogenesis and treatment of ARHL. This paper summarizes the pathogensis and prevention of ARHL.
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