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    Original articles
  • Original articles
    GUO Jiageng, CHEN Liping, YU Yan, SHEN Wei, WU Haitao
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    Objective To investigate the age-dependent modulatory effects of treadmill exercise on spatial learning and memory in mice and to elucidate the potential neurobiological mechanisms underlying these effects. Methods Male C57BL/6J mice at three distinct developmental stages were subjected to a controlled treadmill training protocol for a duration of four weeks. Cognitive performance was assessed in terms of spatial learning and memoryusing a series of behavioral tests. To examine exercise-induced neurogenesis,proliferating adult neural stem cells in the hippocampal region were labeled via intraperitoneal administration of BrdU. Meanwhile,physiological parameters,including body and muscle weight,were monitored throughout the experiment. Results The study revealed significant age-specific effects of treadmill training. In one-month-old mice,exercise intervention markedly enhanced spatial learning and memory,along with increased proliferation of hippocampal neural stem cells,but no significant alterations were observed in body or muscle weight. In six-month-old mice,treadmill training selectively improved spatial memory and led to increased muscle weight. Notably,three-month-old mice exhibited no significant exercise-induced changes in cognitive performance,hippocampal neurogenesis,or muscle weight. Conclusion These findings demonstrate that treadmill exercise exerts significant,age-dependent regulatory effects on spatial learning and memory,with the most pronounced and comprehensive improvements observed in one-month-old mice. The cognitive enhancements may be mediated,at least partially,through exercise-induced promotion of hippocampal neurogenesis. This study provides data for elucidation of the mechanistic basis of exercise-mediated cognitive enhancement and contributes to the potential applications of exercise interventions in cognitive optimization and neuroplasticity across different developmental stages.
  • Original articles
    ZHU Xiaoyu, QIN Tongtong, ZHA Jinhong, ZHOU Dongsheng, YANG Wenhui, GAO Bo
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    Objective To explore the role of Schlafen4 (SLFN4) in acute pneumonia induced by hypervirulent Klebsiella pneumoniae (hvKp) via intratracheal aerosolization. Methods Differential expression gene Slfn4 was identified after infection with hvKp based on RNA sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq) data before Slfn4-/- mice were obtained via CRISPR/Cas gene editing technology. Slfn4-/- mice and wild mice were challenged via intratracheal aerosolization. Mortality and weight changes were recorded for 14 d, while pathological changes and expression levels of interleukin-6 (IL-6), IL-17A, IL-1β, and tumor necrosis factor-α (TNF-α) were detected at 48 h post-infection. Results SLFN4 expression was significantly increased in wild mice after infection with hvKp. Survival was significantly increased, and weight loss was mitigated before gradual recovery in Slfn4-/- mice after infection. The knockout of SLFN4 attenuated alveolar wall thickening, diminished neutrophil infiltration, and suppressed pro-inflammatory cytokine production (IL-6, IL-17A, IL-1β, TNF-α) in the lung at 48 h post-infection. Conclusion The deletion of SLFN4 may suppress the expression of specific pro-inflammatory cytokines and attenuate neutrophil over-recruitment in the lung, thereby alleviating pneumonia in mice after hvKp infection.
  • Original articles
    WU Wenjun, ZHAO Haixin, GAO Jun, WANG Kai, HAN Qiuying, LI Teng, PAN Xin
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    Objective To establish models for real-time dynamic monitoring of intracellular cytoplasmic adenosine triphosphate (ATP) and mitochondrial ATP levels in cells in order to study the changes in metabolic processes. Methods The lentiviral plasmids of the cytoplasmic chemogenetic green fluorescent protein (GFP)ATP probe (Chemo-G)and those of the mitochondrial-localized chemogenetic GFP ATP probe (mito-Chemo-G) were constructed before being transfected into HEK293T together with the helper plasmids, respectively. Chemo-G and mito-Chemo-G lentiviruses were obtained. HeLa cells were infected with the lentivirus. Puromycin resistance selection and flow cytometry cell sorting were employed to identify and isolate the infected HeLa cells. The growth and GFP expressions of HeLa cells were observed. A live-cell imaging system was used for continuous imaging of the cells, with stimuli added at specific time points to alter intracellular ATP levels to observe changes in the fluorescence intensity of the ATP probe. Results Lentiviral plasmids containing Chemo-G and mito-Chemo-G sequences were constructed. Two cell lines which could stably express Chemo-G and mito-Chemo-G were established that exhibited strong growth and accurate intracellular fluorescence localization. Live-cell imaging revealed that after the addition of 2-deoxy-D-glucose (2-DG) into HeLa-Chemo-G, the fluorescence resonance energy transfer (FRET)/GFP ratio showed a decrease that was partially reversed by the addition of glucose. The FRET/GFP ratio increased after histamine stimulation, but rapidly decreased after the addition of oligomycin. Conclusion The Chemo-G and mito-Chemo-G lentiviral vectors and stably transfected cell lines HeLa-Chemo-G and HeLa-mito-Chemo-G are constructed, which provides reliable experimental models for studying cellular metabolism and changes in intracellular ATP levels.
  • Original articles
    ZENG Lingmin, LU Dingyi, CHEN Jiayi, ZHANG Haoqian, GAO Jun, HAN Qiuying, PAN Xin
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    Objective To establish a stable in vitro model of CD8+ T cell exhaustion. Methods CD8+ T cells were isolated and purified from the spleens of ovalbumin-specific CD8+ T cell receptor (OT-I) transgenic mice and subjected to chronic antigen stimulation to induce exhaustion in vitro. Flow cytometry was employed to evaluate the expressions of exhaustion markers, secretion of effector cytokines, and transcription factor profiles in CD8+ T cells. Exhausted and effector (non-exhausted) CD8+ T cells were co-cultured with tumor cells before tumor cell viability was measured to assess the cytotoxic potential of CD8+ T cells. Additionally, N-acetyl-L-cysteine (N-AC) was used as a positive control during exhaustion induction to validate the model. Results Chronic stimulation resulted in a significant upregulation of inhibitory receptors, including programmed cell death protein 1 (PD-1), T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3), T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motifdomain (TIGIT), and lymphocyte activation gene 3 (LAG-3). Concurrently, the secretion of key effector cytokines such as tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) was markedly reduced. Exhausted CD8+ T cells exhibited diminished cytotoxicity against tumor cells compared to effector CD8+ T cells. Notably, treatment with N-AC effectively restored the function of exhausted CD8+ T cells and enhanced their anti-tumor activity. Conclusion This study has established an effective in vitro model for CD8+ T cell exhaustion. The use of N-AC demonstrates its potential to restore functionality in exhausted CD8+ T cells, underscoring the reliability and utility of this model for investigating the anti-tumor potential of exhausted T cells.
  • Original articles
    LIU Yinji, DU Chunxiao, CHEN Junrui, LI Yuxiang, LI Ge, WANG Zhiding, HAN Gencheng
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    Objective To investigate the effects of music therapy on cognitive function and immunity of sleep-deprived mice and the potential underlying mechanisms. Methods A novel music therapy was developed by integrating elements from both Western and Chinese music. A sleep-deprived mouse model was established to explore the effects of the music combination on learning and memory of mice using Morris water maze experiments. ELISA was used to detect immune-endocrine indicators in the blood and saliva of mice and to study the effects of this music combination on IgA levels.Transcriptome high-throughput sequencing and B-cell receptor (BCR) repertoire sequencing were adopted to explore the potential mechanisms through which music therapy influenced IgA production. Results The Morris water maze test revealed that the novel music therapy could promote the recovery of cognition and memory of sleep-deprived mice. Additionally, it was found that the music combination could increase IgA levels in both blood and saliva. Transcriptome high-throughput sequencing and BCR sequencing analysis showed that the music combination enhanced the abundance of the IgA immunoglobulin light chain variable region (Igkv4-53) and heavy chain constant region (Igha). Conclusion Music therapy can help restore cognitive function and increase IgA levels in sleep-deprived mice. The mechanism may be related to the enhanced abundance of immunoglobulin light chain variable region (Igkv4-53) and heavy chain constant region (Igha).
  • Original articles
    XIE Hao, AI Lingli, SU Duo, LI Lu, HU Lingfei, ZHOU Dongsheng, GAO Bo, YANG Huiying
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    Objective To explore the biological functions of pyroptosis-related genes in pneumonic plague using bioinformatics methods, and to evaluate their potential applicability as diagnostic markers. Methods The pneumonic plague-related dataset GSE220123 was retrieved from the Gene Expression Omnibus (GEO) database and screened for differentially expressed pyroptosis-related genes (DE-PRGs). The functions of DE-PRGs were studied via Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and immune infiltration analysis. The hub genes were identified via protein-protein interaction (PPI) network analysis, and further screened for key genes with sustained high expression characteristics based on differential expression analysis.The relative expression levels of the key genes were verified using the reverse transcription real-time quantitative PCR (qPCR) method. Results A total of 17 DE-PRGs were screened, and PPI network analysis revealed 7 Hub genes. Among them, Casp4 continued to be up-regulated during the course of pneumonic plague. The results of reverse transcription qPCR were consistent with the those of bioinformatic analyses. Conclusion DE-PRGs play a crucial role in the immune response of pneumonic plague, especially Casp4, which has significant applications as a diagnostic biomarker and potential therapeutic target for pneumonic plague.
  • Original articles
    YE Zhiyong, YU Xianxing, JIANG Hui
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    Objective To investigate the applicability of D-dimer levels combined with the Tei index in assessing the volume load in patients with acute decompensated heart failure (ADHF). Methods Two hundred patients admitted to the Emergency Department of Taizhou TCM Hospital between 2022 and 2023 were selected as the subjects, who were divided into a low volume load group (n=121) and a high volume load group (n=79) based on plasma volume indices.The clinical indicators and parameters of cardiac function of these subjects were compared. Multivariate Logistic regression analysis was used for identification of potential risk factors for elevated volume loads.A model for prediction of elevated volume loads was developed via Chi-square automatic interaction detection, followed by internal validation via Bootstrapping. Logistic regression analysis was used to study the interactions between D-dimer levels and Tei indices in assessing the volume loads of patients with ADHF. Results Multivariate regression analysis showed that age, Tei index, D-dimer levels and B type natriuretic peptide(BNP) were risk factors for high volume loads in ADHF patients. The decision tree model established had 14 nodes and 8 terminal nodes. Four explanatory variables including age, Tei index, D-dimer levels and BNP were screened out. The area under curve(AUC), sensitivity, specificity and prediction accuracy of the classification tree model were higher than those of the regression model. When D-dimer levels ≥1.16 and Tei index ≥0.65 interacted and co-existed, ADHF patients were at higher risk of developing high volume loads (OR=6.526, 95%CI: 2.439-7.183). The results of restricted cubic spline analysis pointed to nonlinear dose response relationships in the correlation strength of D-dimer levels, Tei index and volume loads. Conclusion There are interactions between D-dimer levels and Tei index, and the risk of high volume loads in ADHF patients is significantly increased when D-dimer levels are 1.16 or above and Tei index is 0.65 or above.
  • Original articles
    DONG Yutong, YANG Maolin, LI Mo, ZHANG Chen, RUAN Yan, ZHANG Junlei, HU Yan
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    Objective To investigate the differences in physical fitness and physiological indexes of male soldiers stationed at different altitudes and to analyze the determinants of physical fitness of these soldiers. Methods A total of 13 648 male soldiers at different altitudes were included, whose results of annual military physical assessment and physiological indexes in 2023 were collected and comparatively analyzed. Results The 3000-meter running performance, 30-meter × 2 serpentine running performance and resting blood oxygen saturation of high-altitude soldiers gradually declined with the increase of altitudes. The pull-up performance dropped significantly when the altitude >2000 m. Soldiers ≥30 years old demonstrated a marked decline in physical fitness across different training programs. Moderate altitude acclimatization was conduciveto the recovery of physical fitness of soldiers,while exposure to high-altitude environment ≥5 years led to overall deterioration. Conclusion The high-altitude environment significantly impacts the aerobic endurance, speed, agility, and strength of soldiers. Altitude ≥4000 m, age ≥30 years, and prolonged exposure ≥5 years may contribute to the decline in physical fitness of soldiers.
  • Original articles
    TIAN Wenxue, GAO Shen, RONG Xiangyun, ZHANG Zhaoxue, CAI Jianming, SHENG Fugeng, ZHANG Hongtao
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    Objective To investigate the effect of T2-fluid attenuated inversion recovery(T2-FLAIR) sequence on the appearance of ivy sign in patients with moyamoya disease under different imaging conditions. Methods Coronal T2-FLAIR scans were performed in 51 consecutive adult patients with moyamoya disease who had not undergone surgery and had their first visit to this hospital between March and July of 2024. According to the scanning conditions, the patients were divided into 19 and 14 of the echo train length in two groups,and 103, 144, and 195 ms of the time of echo(TE) in three groups, respectively. The left and right cerebral hemispheres and whole brains were scored according to the cerebral vascular anatomy, and scores of the ivy sign of left and right cerebral hemispheres and whole brain were compared. Results There was no statistical significance in the ivy sign scores of right and left cerebral hemispheres and whole brain between the two groups with 19 and 14 of the echo train length (P>0.05). Comparison of ivy sign scores in right and left cerebral hemispheres and whole brain was statistically significant among the three groups of TE at 144, 103 and 195 ms (P<0.05). Conclusion The best appearance of ivy sign in patients with moyamoya disease is seen under the condition of TE at 195 ms,so appropriately extending the TE time is helpful for ivy sign display.
  • Reviews
  • Reviews
    ZHANG Haoqian, PAN Xin, QIAO Yue, XUAN Chengrui
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    Metabolic reprogramming in the tumor microenvironment is closely associated with tumor initiation and progression. Endogenous metabolites can not only regulate tumor formation and metastasis, but also impair antitumor immune efficacy by influencing immune cells. However, current research focuses on the effects of single metabolites on tumors rather than take into account the systemic nature of metabolic networks, the diversity of metabolic products, and their dynamic regulatory roles in the tumor immune microenvironment. An in-depth analysis of the key nodes in metabolic pathways related to endogenous metabolites and their regulatory mechanisms in tumor progression is crucial to the development of antitumor therapies based on metabolic intervention. This paper provides an overview of the molecular mechanisms by which endogenous metabolites, such as amino acids, lipids and nucleotides, contribute to tumor progression and regulate the immune microenvironment. By offering new theoretical insights into the complex network of the metabolism-immune axis in tumor development, this review aims to provide data for the development of metabolism-targeted antitumor immunotherapy strategies.
  • Reviews
    YANG Yanjiao, GAO Weina
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    Pyroptosis is a programmed cell death accompanied by cell swelling, nuclear condensation, membrane rupture, and the release of intracellular inflammatory factors. Pyroptosis is closely related to the occurrence and development of multiple organ and system diseases, and can trigger a cascade of inflammatory responses. Studies have demonstrated that effective components of traditional Chinese medicine (TCM) exhibit significant effects on modulating pyroptosis and the subsequent inflammatory responses. Here we review the research progress in studies on how TCM effective components improve diseases via pyroptosis in order to provide data for the development of new TCMs.
  • Short report
  • Short report
    LIN Huiyan, WANG Huan, GU Xiaofeng, WU Yutian, YUAN Kang, LI Feifei, SHA Ning, WANG Hui, LOU Yunpeng
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