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  Original articles

  A CRISPR-Cas9 mediated technology for controlling the bla_NDM-1 gene

  SHAO Yutong, JIN Min, YANG Dong

  2026, 50(8): 561-569. https://doi.org/10.7644/j.issn.1674-9960.2026-00032

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  Objective To construct a single-plasmid clustered regularly interspaced short palindromic repeats（CRISPR）/CRISPR-associated nuclease 9（Cas9）single-plasmid system integrated with conjugative transfer elements, evaluate its ability to eliminate antibiotic resistance genes （ARG） harbored by bacteria, and explore the feasibility of containing the dissemination of ARG via conjugative transfer. Methods A CRISPR-Cas9 plasmid vector targeting the bla_NDM-1 gene was designed, and the oriT-traJ conjugative transfer module was integrated into the vector backbone. Electrotransformation and conjugative transfer assays were performed to assess the elimination efficiency of this system against bla_NDM-1. Antibiotic susceptibility assays were conducted to verify the restoration of antibiotic susceptibility in edited bacterial strains. Results The elimination efficiency of the established CRISPR-Cas9 single-plasmid system was 97.9%, 87.5% and 56.3% against bla_NDM-1 located on the chromosome, plasmid, and dual location respectively. PCR results demonstrated that the system was still able to eliminate target genes after conjugative transfer and efficiently reversed the antibiotic resistance of recipient bacteria. Conclusion This system can eliminate the bla_NDM-1 gene and restore bacterial susceptibility to meropenem. Conjugative transfer delivery mediated by the single-plasmid system provides a novel tool for eradicating ARG and restoring the susceptibility of recipient bacteria to antibiotics.
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  Original articles

  Isolation, short-term culture and characterization of storage cells in Hypsibius henanensis

  WANG Yifan, LI Lei, ZHANG Lingqiang

  2026, 50(8): 570-578. https://doi.org/10.7644/j.issn.1674-9960.2026-00045

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  Objective To establish a method for isolation and short-term culture of storage cells of Hypsibius henanensis, characterize their features, and evaluate their radiation tolerance so as to provide an in vitro platform for investigating stress-resistance mechanisms of tardigrades. Methods Laboratory-cultured H.henanensis was used as the subject. Storage cells were mechanically isolated. The osmotic pressure of the tardigrade homogenate was measured to optimize the culture conditions. The isolated cells were maintained in a short-term culture using modified Grace’s medium. The cell viability was determined via trypan blue exclusion and calcein acetoxymethyl ester (Calcein-AM) and propidium iodide (PI) double staining. The morphology and components of these cells were characterized by fluorescence staining. Radiation tolerance was evaluated by comparing the survival rate of storage cells with that of HeLa cells after exposure to 500 Gy of X-ray irradiation. Results High-purity storage cells were isolated, which were round or polygonal in shape with an average diameter of (7.50±0.15) μm. These cells had intact cell membranes and contained abundant cytoplasmic neutral lipid droplets, with mitochondria evenly distributed. The optimized culture system could keep cell viability stable for 7 d. After 500 Gy X-ray irradiation, the survival rate of storage cells was (76.30±17.32)%, which was not significantly different from that of the non-irradiated group (74.71±14.74%)(P>0.05), unlike HeLa cells whose survival rate decreased significantly (P<0.01). Conclusion A method for isolation and short-term culture of storage cells from H.henanensis has been established. These storage cells are abundant in lipid reserves and capable of radiation tolerance in vitro. These findings can help construct cell models and study the stress-resistance mechanisms of tardigrades.
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  Original articles

  Cloning, identification and anti-radiation function of HhGATA3 gene from Hypsibius henanensis

  LIU Yanyan, LI Lei, ZHANG Lingqiang

  2026, 50(8): 579-589. https://doi.org/10.7644/j.issn.1674-9960.2026-00047

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  Objective To clone and characterize Hypsibius henanensis GATA binding protein 3 （HhGATA3） and to investigate its function in radiation tolerance. Methods The coding sequence of HhGATA3 was amplified from H.henanensis cDNA by PCR, and the protein was analyzed using bioinformatics tools.The subcellular aggregation of HhGATA3 was observed by immunofluorescence.The radioprotective capacity was evaluated via alkaline comet assay and phosphorylated histone H2AX （γH2AX） foci formation in human cells expressing HhGATA3.The knockdown efficiency of small interfering RNA （siRNA） was verified by reverse transcription real-time quantitative PCR （qPCR） in H.henanensis, and the post-irradiation survival rate was measured. Results The HhGATA3 gene was cloned.HhGATA3 protein formed phase-separated droplets within cells.Comet and γH2AX foci assays suggested that HeLa cells expressing HhGATA3 had their DNA damage significantly mitigated after X-ray irradiation （P<0.0001）. Knockdown of HhGATA3 in H.henanensis markedly decreased the post-irradiation survival rate to approximately 40% （P<0.001）. Conclusion HhGATA3 plays a critical role in the radiation tolerance of H.henanensis. In human HeLa cells, HhGATA3 can undergo phase separation and mitigate radiation-induced DNA damage.
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  Time-and dose-effect relationships of DNA adducts formation induced by nitrogen mustards in Neuro-2a cells

  LIU Xiaohui, WANG Zhaoxia, LIU Yulong, WU Jianfeng, CHEN Aibing, XU Hua, XIE Jianwei

  2026, 50(8): 590-598. https://doi.org/10.7644/j.issn.1674-9960.2026-00049

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  Objective To identify the biomarkers of effect in DNA damage induced by nitrogen mustard treatment. Methods Neuro-2a cells were subjected to in vitro treatment with nitrogen mustards （HN1, HN2 or HN3）. The types and levels of HN-DNA adducts were quantitatively analyzed using the multiple reaction monitoring （MRM） mode coupled with ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）. Results Good linearity was observed in the nine HN-DNA adduct standards within the range of 0.02 to 100 ng/mL, with coefficients of determination above 0.99. The limits of detection （LOD） and limits of quantitation （LOQ） ranged from 0.01 to 0.02 ng/mL and from 0.02 to 0.05 ng/mL, respectively. The relative standard deviations （RSD） for intra-assay and inter-assay precision were less than 15%. Such adducts as HN-N3-adenine（AN3）, HN-N7-guanine（GN7）, and HN-Bis-GN7 were detected in cells treated with the three nitrogen mustards. Adduct levels increased immediately after treatment and decreased 8 to 12 h later, showing a positive correlation with the dose of treatment. Conclusion By dynamically monitoring the temporal and quantitative changes of DNA adducts in Neuro-2a cells, this study confirms their potential as effect biomarkers in nitrogen mustard treatment while providing data for investigating the repair processes and mechanisms of DNA damage induced by nitrogen mustards.
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  Original articles

  Preventive effect of Morinda officinalis oligosaccharides against post-traumatic stress disorder

  ZHANG Xiang, LI Zhihan, WANG Zhiding, LI Yunfeng

  2026, 50(8): 599-607. https://doi.org/10.7644/j.issn.1674-9960.2026-00068

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  Objective To investigate the preventive effect of inulin-type oligosaccharides of Morinda officinalis （IOMO） against post-traumatic stress disorder （PTSD） like behaviors induced by inescapable electric foot-shock （Fs） in mice and the underlying mechanism. Methods A mouse model of PTSD was established using inescapable electric Fs. Mice were randomly divided into the control group, Fs group, Fs+sertraline （Ser） positive drug group, and Fs+IOMO prevention group. The IOMO group was administered with 50 mg/kg IOMO once daily by intragastric gavage （ig） for 10 consecutive days before modeling. PTSD like behaviors were evaluated using the contextual fear paradigm （CFP）, open field test （OFT）, and elevated plus maze （EPM）. Gut microbiota composition was analyzed by 16S rRNA sequencing. Pathological changes in colon tissue were observed by HE staining. Levels of lipopolysaccharide （LPS）, interleukin-6 （IL-6）, interleukin-1β （IL-1β）, and tumor necrosis factor-α （TNF-α） in the serum and hippocampus were measured by enzyme-linked immunosorbent assay （ELISA）. Protein expressions of Claudin 1 in colon tissue and NOD-like receptor thermal protein domain associated protein 3 （NLRP3） in the hippocampus were detected by Western blotting. Results Compared with the control group, mice in the Fs group exhibited significant PTSD like behaviors without affecting spontaneous activity, as evidenced by prolonged freezing time in the CFP, prolonged latency to enter the intermediate zone and reduced percentage of distance traveled in the intermediate zone in the OFT, and prolonged latency to enter the open arm and reduced percentage of distance traveled in the open arm in the EPM. Preventive administration of IOMO for 10 days before modeling significantly reversed these PTSD like behaviors. Meanwhile, significant disturbance of gut microbiota composition was observed in the Fs group, which was markedly restored by IOMO pretreatment. In the Fs group, colon Claudin-1 expression was decreased, while serum and hippocampal LPS levels as well as hippocampal NLRP3 and inflammatory cytokine levels were significantly increased. IOMO significantly reversed these changes. Conclusion Preventive administration of IOMO can effectively prevent PTSD like behaviors in mice. This effect may be associated with regulation of gut microbiota, enhancement of intestinal barrier function, and alleviation of hippocampal neuroinflammation. This study provides correlative evidence for the gut-brain axis mediating the preventive effect of IOMO against PTSD.
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  Original articles

  Spectral characteristics of functional near-infrared imaging under different levels of attention

  GONG Jiahui, XU Donggang, FU Wenliang, ZHANG Chao, XIA Wenrong, ZHAO Yongqi, GAO Bo, XING Weiwei, ZUO Zhongfu

  2026, 50(8): 608-613. https://doi.org/10.7644/j.issn.1674-9960.2025-00117

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  Objective To investigate the characteristics of brain activation in the prefrontal and parietal cortices under varying attention levels using functional near-infrared spectroscopy （fNIRS） in order to provide data for real-time attention monitoring. Methods Hemodynamic responses of oxygenated hemoglobin were recorded from 45 healthy participants during the psychomotor vigilance task （PVT）. The fastest 10% of reaction times were defined as the high-attention state while the slowest 10% were defined as the low-attention state. Changes in oxyhemoglobin concentration （ΔHbO） and their time-domain parameters were compared between these states. Receiver operating characteristic （ROC） curves were used to evaluate the diagnostic value. Results The dorsolateral prefrontal cortex,primary motor cortex,and premotor cortex showed significantly stronger activation in the high-attention state. Analysis of characteristic parameters revealed that integral values,linear slopes and centroid values were higher in the high-attention state than in the low-attention state. Notably,the integral value of the dorsolateral prefrontal cortex was moderately accurate in monitoring attention（AUC=0.710）. Conclusion The integral value of oxyhemoglobin in the dorsolateral prefrontal cortex may serve as a potential biomarker for attention assessment. This finding offers key parameters for real-time,dynamic attention monitoring using fNIRS.
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  Original articles

  Design and implementation of a health management information system for radiation staff

  ZHANG Huiqiang, YAN Li, GAO Ning, HAN Wenhui, ZHAO Han, LI Zunting, WU Yitong, YAN Tao, ZHANG Peng, WANG Junliang, YU Changlin, YANG Xinglong, YANG Wenfeng, WANG Jianying, SU Jianbin

  2026, 50(8): 614-618. https://doi.org/10.7644/j.issn.1674-9960.2025-00237

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  Objective To develop a health management information system for radiation staff so as to upgrade the informatization and standardization of radiation protection and safety management, and provide technical support for the occupational health of the staff concerned. Methods The system design was completed by referring to national laws, regulations and standards, conducting on-site investigations, and consulting experts. The system was based on a microservice architecture, Java EE technical specifications and in the Browser/Server （B/S） mode. The scheme of the system was optimized and improved via verification by and feedback from radiation-related departments. Results The demands of three dominating stakeholders-the radiation department, radiation staff, and medical assessment experts-were clearly defined. A framework of applicationsthat was regulated at different levels, one for standardized technologies, and a third one for the whole-process functions were constructed. The system was equipped with 12 core functional modules involving personal information, workability of radiation facilities, dose monitoring, health care, intelligent prompt and early warning, and statistical analysis, which was capable of full-cycle digital management of occupational health of the radiation staff. Conclusion The system can well meet the management requirements of special industries such as the military and the refined management needs of grassroots institutions. This system can help diversify the means of informatized management of occupational health of the radiation staff, and can provide references for related departments.
Reviews
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  Reviews

  Cross-population applications of stroboscopic visual training： implementation strategies, effects, and potential mechanisms

  JIANG Dongting, GUO Zhenxiang, WANG Xing, YANG Yi, LING Xiaoyu

  2026, 50(8): 619-627. https://doi.org/10.7644/j.issn.1674-9960.2026-00041.2026-00041

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  Stroboscopic visual training （SVT） is a neurocognitive intervention that intermittently occludes visual input, thereby promoting adaptive modulation of the central nervous system under heightened perceptual load. In recent years, SVT has been increasingly applied in athletic training, health promotion, and clinical rehabilitation, demonstrating multidimensional intervention potential across diverse populations. This review systematically summarizes the implementation strategies and training effects of SVT in elite athletes, healthy adults, individuals with balance impairments, and older adults, with a focus on its effects on cognitive processing, motor control, and sensory integration. Current evidence suggests that SVT protocols should be individually tailored according to stroboscopic parameters, such as frequency and duty cycle, as well as the overall training period, with intervention effects varying substantially across populations. SVT may enhance visual perception and cognitive processing efficiency in athletes, strengthen cognitive control and postural stability in healthy adults, and promote sensory compensation and motor function remodeling in individuals with neurocognitive disorders or musculoskeletal injuries. Mechanistically, SVT may optimize perception-action integration by improving visual information processing efficiency, reshaping attentional strategies, enhancing predictive processing, and facilitating sensory reweighting. Future studies should further examine dose-response relationships in older adults and rehabilitation populations, and use neuroimaging techniques to clarify the underlying central mechanisms.
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  Reviews

  Advances in the preparation and clinical application of freeze-dried platelets

  ZHANG Ruotong, FU Qiuxia, YI Xiaoyang

  2026, 50(8): 628-634. https://doi.org/10.7644/j.issn.1674-9960.2026-0011

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  Freeze-dried platelets （FDPs）, characterized by long-term storage at the ambient temperature, convenient transport and pathogen inactivation, are extensively studied in transfusion and regenerative medicine. This review summarizes the whole preparation procedures of FDPs including pretreatment, pre-freezing, lyophilization and rehydration, compares four rehydration media, and reviews FDP applications in thrombocytopenia transfusion, traumatic shock rescue and tissue repair. FDPs exhibit favorable hemostatic activity and biocompatibility, but they are vulnerable to lyophilization-induced structural damage, weak in vitro aggregation function, inadequate in clinical trials and unified quality control standards. Future research will focus on developing compound protectants, standardizing preparation workflows, clarifying molecular mechanisms of lyophilization injury via multi-omics, and establishing unified quality specifications through multicenter clinical trials. Optimized preparation technologies of FDPs will revolutionize the way platelets are stored and transported so as to provide a novel blood product for emergency hemostasis and wound repair.
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  Reviews

  Research progress in soft mist inhalers and challenges

  REN Bo, WANG Libin, LIU Chunyuan, LIU Chunyan, ZHENG Aiping

  2026, 50(8): 635-640. https://doi.org/10.7644/j.issn.1674-9960.2025-0027

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  Given the unique physiological structure of lungs, pulmonary drug delivery has distinct advantages over oral or injectable administration routes for treating both pulmonary and systemic diseases, hence increasing interest in related research. Despite the ability to achieve pulmonary drug delivery, conventional inhalable formulations have such limitations as low rates of pulmonary deposition, a lack of user-friendliness, relatively high carbon emissions, and poor patient compliance. As a novel inhalation device, soft mist inhalers （SMIs） promise good potential and advantages. This article outlines the aerosol characteristics and principles of SMIs, summarizes their origins, leading advantages, and clinical value, and analyzes its applications in shear-sensitive formulations based on related literature and patents. Recent representative patents related to SMIs are listed. Furthermore, the challenges facing the development of SMIs are discussed, involving the optimization of drug solution formulations, enhancement of nozzle design precision, and reduction of production costs. Future developments are also predicted.