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  Original articles

  Mutations and functional mechanisms of deubiquitinating enzyme OTU domain-containing protein 3 in colorectal cancer

  ZHANG Jinhao, ZHANG Yonghui, LI Hongchang, ZHANG Lingqiang

  2025, 49(5): 321-329. https://doi.org/10.7644/j.issn.1674-9960.2025.05.001

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  Objective To investigate mutations in OTU domain-containing protein 3（OTUD3） and their functional mechanisms in the initiation and progression of colorectal cancer（CRC）. Methods Gene expression profiling interactive analysis （GEPIA2） and the human protein atlas database （THPA） were used to analyze gene transcription and protein expressions. Samples from 32 patients with CRC were collected to identify OTUD3 mutants. Based on the information about mutation sitesof OTUD3 in an existing database, a plasmid vector containing the OTUD3 gene mutant was constructed. Plasmid vectors containing the phosphatase and tensin homolog deleted on chromosome 10 （PTEN） and the OTUD3 gene mutant were co-transfected into HCT116 cells. Western blotting, half-life, immunoprecipitation, ubiquitination, and hybrid algorithm molecular docking （H-DOCK） assays were employed to find out whether and why the OTUD3 mutants affected PTEN protein levels.Functional alterations in CRC cells after OTUD3 mutation were verified by CCK-8 cell proliferation, transwell cell invasion, scratch, and clonal formation assays. Results OTUD3 mutations were highly frequent in CRC. OTUD3 mutants R178W and N321S resulted in the loss of function of the stable PTEN protein, leading to enhanced proliferation, invasion, migration, and survival of CRC. Conclusion In CRC, OTUD3 mutation reduces the ability to stabilize PTEN and promotes the occurrence and development of CRC.
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  Original articles

  Deoxycholic acid mediates neuroinflammatory responses under hypoxia exposure

  CUI Haoran, XING Chen, SONG Lun

  2025, 49(5): 330-336. https://doi.org/10.7644/j.issn.1674-9960.2025.05.002

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  Objective To investigate the roles and mechanism of deoxycholic acid （DCA） in mediating the neuroinflammatory responses induced by hypobaric hypoxia. Methods C57BL/6J mice were randomly divided into thecontrol group （Con）, tauroursodeoxycholic acid （TUDCA） group, hypoxia group （Hyp） and hypoxia+TUDCA group （Hyp+TUDCA）. The mice were exposed in a hypoxic chamber for 48 h, which could mimic the high altitude environment. The levels of total bile acids （BA）, lithocholic acid （LCA） and deoxycholic acid （DCA） in the serum and colon of mice were detected using enzyme-linked immunosorbent assay （ELISA）, so were levels of DCA in the cortex, hippocampus and hypothalamus. The role of DCA in inducing neuroinflammation and IL-1β and TNF-α expressions was evaluated by knocking down the DCA receptor, farnesoid X receptor （FXR） in the N9 microglial cells and by administrating TUDCA to mice followed by hypoxia exposure. Real-time quantitative PCR （qPCR） and immunofluorescence staining were used to detect the activation of microglia and expression levels of interleukin-1β（IL-1β） and tumor necrosis factor-α （TNF-α） in the cortex of mice in each group. Results Hypoxia exposure increased DCA levels in the colon, serum and cortex. Under the same conditions, cortical microglia were activated, along with the elevation of IL-1β and TNF-α levels. DCA treatment increased the expression levels of FXR, IL-1β and TNF-α in N9 microglial cells. Knocking down the expression level of FXR inhibited DCA-induced IL-1β and TNF-α expressions in N9 cells. Furthermore, administration of TUDCA inhibited microglial activation and elevation of IL-1β in the cortex induced by hypoxia. Conclusion DCA can serve as a mediator in neuroinflammation by activating FXR in the cortex under hypoxia exposure. TUDCA administration can be used to mitigate neuroinflammation induced by hypoxia.
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  Original articles

  A research on tissue damage detection based on mutations in cell-free DNA

  LAN Shishi, HUANG Ye, WANG Chunhui, ZHANG Hongxing

  2025, 49(5): 337-348. https://doi.org/10.7644/j.issn.1674-9960.2025.05.003

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  Objective To investigate the feasibility of using the cell-free DNA （cfDNA） carrying tissue-specific mutations as biomarkers for assessing the severity of exercise-induced tissue injury. Methods Based on a public gene expression database, ten tissue-specific and highly-expressed genes were selected for each of ten different human tissues. A total of 34 young healthy volunteers were recruited, and their peripheral blood samples were collected after running 5 km per day for one week. Genomic DNA from leukocytes, plasma exosomal RNA, and plasma cfDNA were extracted and subjected to high-throughput sequencing respectively. Tissue-specific somatic mutations in plasma exosomal RNA were identified, and the relationship between cfDNA carrying these mutations and traditional protein biomarkers was analyzed. Results Tissue-specific cfDNA mutations related to five tissues （myocardium, skeletal muscle, intestine, stomach, and kidney） were detected in more than five volunteers. The correlation coefficients between total plasma cfDNA levels and the levels of tissue-specific protein biomarkers associated with these five tissues were less than 0.3 （n=34, r=-0.51-0.28, P=0.0022-0.65）. Notably, for each specific tissue, the levels of tissue-specific cfDNA mutations were positively correlated with the corresponding protein biomarker levels, and correlation coefficients were over 0.8 （n=7-13, r=0.81-0.92, P=9.0× 10^-4-0.020）. Conclusion Tissue-specific cfDNA mutations may serve as novel potential biomarkers for non-invasive evaluation of exercise-related tissue injury.
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  Preparation and toxicity analysis of recombinant Clostridium perfringens alpha toxin

  ZHAO Pinnan, GAO Xiang, LUO Longlong

  2025, 49(5): 349-355. https://doi.org/10.7644/j.issn.1674-9960.2025.05.004

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  Objective To efficiently produce Clostridium perfringens alpha toxin （CPA） by using a prokaryotic expression system and systematically analyze its biological activity. Methods The CPA gene fragment fused with His-Tag sequence was cloned into a prokaryotic expression vector pTIG-Trx and transformed into E. coli TransB （DE3）. The IPTG was used to induce the expression of the target protein,and the protein was obtained by using Ni-column affinity purification. The cytotoxic effect of recombinant CPA protein on 293T,LS174T and SW480 cells,hemolytic effect on human and mouse red blood cells,and lethal effect on mice were further evaluated. Results The recombinant CPA protein with a relative molecular weight of about 45×103 and a purity of more than 90% was successfully obtained. It had significant toxicity to 293T,LS174T and SW480 cells and induced hemolytic reactions in human and mouse red blood cells at specific concentrations. Low dose of CPA protein could cause rapid death in mice in a short time. Conclusion This study successfully obtained the high purity CPA protein with good biological activity in vitro and in vivo,which laid a foundation for further study of the pathogenic mechanism of CPA and its potential application.
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  Drug target discovery for idiopathic pulmonary fibrosis via druggable genome-wide Mendelian randomization

  LIN Xueyang, LANG Simin, YANG Yufeng, YANG Chen, CUI Ziqi, LUO Yuan, WANG Yongan

  2025, 49(5): 356-363. https://doi.org/10.7644/j.issn.1674-9960.2025.05.005

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  Objective To identify potential drug target genes associated with idiopathic pulmonary fibrosis （IPF） and predict therapeutic candidates using a two-sample Mendelian randomization （MR） approach across the druggable genome. Methods Druggable genome data from the DGIdb database and Finan were integrated to identify overlapping genes. A two-sample MR analysis was performed to infer causal relationships between genes and IPF. Functional enrichment analyses,including Gene Ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG）,were conducted to explore biological pathways. Drug-target interactions were predicted via DSigDB database screening,followed by molecular docking simulations to evaluate binding affinities. Results Among the 2588 overlapping druggable genes,thirty exhibited significant causal associations with IPF （P< 0.05）. Four hub genes （NOD2,LATS2,LTA,and TCF7L2） were enriched in IPF-related pathways,notably Hippo and TNF signaling. Six potential therapeutics were identified： oxyphenbutazone,moexipril,α-galactosylceramide,GSK429286A,CGP74514A,and JW-7-24-1. Molecular docking confirmed strong binding affinities between these drugs and their targets. Conclusion This study has identified thirty druggable gene targets and six candidate drugs for IPF. The enrichment of hub genes in key pathways and validated drug-target interactions provide insights into IPF therapies.;
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  Clinical efficacy of Quhan Tongluo Formula combined with celecoxib against knee osteoarthritis

  ZHANG Zhenhua, HUANG Jianghai, WANG Yafei, LI Yunhai

  2025, 49(5): 364-369. https://doi.org/10.7644/j.issn.1674-9960.2025.05.006

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  Objective To investigate the clinical efficacy and safety of Quhan Tongluo Formula in treating knee osteoarthritis（KOA） of cold-dampness obstruction pattern. Methods A retrospective study was conducted on 231 patients with KOA of cold-dampness obstruction pattern admitted to Dongfang Hospital,Beijing University of Chinese Medicine from December 2022 to December 2023. The patients were divided into an exposed group （105 cases） and a non-exposed group （126 cases） based on whether Quhan Tongluo Formula was applied. The exposed group received Quhan Tongluo Formula combined with celecoxib,while the non-exposed group received only celecoxib. Using propensity score matching based on the baseline data of the medical records,76 pairs （a total of 152 cases） were matched. The changes in Visual Analog Scale （VAS） scores,Western Ontario and McMaster Universities Osteoarthritis Index （WOMAC） total scores,and traditional Chinese medicine （TCM） syndrome scores before treatment and at 2,4,and 6 weeks after treatment were compared between the two groups,and safety was evaluated. Results Before treatment,there were no significant differences in VAS scores or WOMAC pain scores between the two groups. After treatment,both groups exhibited significant reductions in VAS scores and WOMAC pain scores,with the exposed group demonstrating significantly greater reductions compared to the non-exposed group （P< 0.01）. The total clinical effective rate was significantly higher in the exposed group than in the non-exposed group （P< 0.01）. Conclusion The clinical application of Quhan Tongluo Formula combined with celecoxib demonstrated superior efficacy over celecoxib monotherapy in treating knee osteoarthritis of cold-dampness obstruction pattern. The combined regimen significantly reduced VAS scores,total WOMAC scores,and TCM syndrome scores,with marked improvements in long-term efficacy.
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  Original articles

  Determination of residual solvents in amidoxime EN chelating agents by headspace gas chromatography

  ZHU Jinshou, XIAO Bingkun, MIAO Xiaoyao, YANG Fang, HUANG Rongqing

  2025, 49(5): 370-373. https://doi.org/10.7644/j.issn.1674-9960.2025.05.007

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  Objective To establish an analytical method for the determination of methanol and ethanol solvent residues in amidoxime EN chelating agents by headspace gas chromatography. Methods The samples were analyzed in a GC-2030 gas chromatograph system using a Shimadzu HS-20 fully automated headspace injector and quantified by an internal standard method. The samples were separated and detected by the DB-WAX capillary column with a flame ionization detector（FID）. The thermostatic furnace temperature was 70 ℃ and the equilibrium time of the sample bottle was 30 min. The sample flow path temperature was 150 ℃ and the carrier gas was high pure nitrogen. As temperature programming, the initial column temperature of 40 ℃ lasted 9 min and increased to 200 ℃ at 40 ℃/min for 2 min. Results The peak areas of the residual solvents in EN chelator showed good linearity （R2> 0.999） in the mass concentration range investigated,the detection limits of solvent methanol and ethanol were 0.25 µg/mL and 0.30 µg/mL respectively, and the limits of quantification were 1.25 µg/mL and 1.50 µg/mL, respectively. The recovery of residual solvent addition ranged from 97.10% to 102.23%, and the RSD value was less than 5.00%. Conclusion This method is sensitive enough to be used for the determination of solvent residues in amidoxime EN chelating agents.;
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  Original articles

  A simulated model for medical evacuation amid mass casualties and

  LUO Jia, CHEN Yao

  2025, 49(5): 374-381. https://doi.org/10.7644/j.issn.1674-9960.2025.05.008

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  Objective To establish a simulated model that can optimize strategies for medical evacuation in case of mass casualties,and to study the guidelines for centralized scheduling of evacuation vehicles. Methods A simulated model was constructed based on theories about discrete event simulation and on processes of medical evacuation. Evacuation processes under different scheduling strategies were simulated and quantitatively evaluated in terms of efficiency of evacuation and utilization of resources.The impact of key parameters on the medical evacuation system was analyzed. Results When the critically wounded were given priority,the rate of immediate treatment for the wounded was the highest（87.44%）,and the median waiting time for the wounded was 23.70 min. In terms of utilization of resources,the strategy of prioritizing the critically wounded delivered the best result,with an average of 2.94 persons evacuated per vehicle and an overall rate of vehicle utilization was 50.93%. The daily number of the wounded and the number of evacuation vehicles made a big difference in the performance of the rescue system. Command and scheduling had to be dynamically optimizedas required so as to strike a balance between the efficiency of treatment and utilizationof resources. Conclusion This simulated model for medical evacuation in case of mass casualties,which is based on theories about discrete event simulation,is flexible enough toback up the simulation of strategy optimization.
Reviews
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  Reviews

  Research progress in prehospital pain management in the U.S. Armed Forces

  WANG Hui, OUYANG Ling, XIE Xiaotong, ZHOU Lingjun

  2025, 49(5): 382-385. https://doi.org/10.7644/j.issn.1674-9960.2025.05.009

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  Pain management is an important component of military medical support during wartime.Early pain management for wounded soldiers can effectively alleviate pain, which is critical to maintaining combat effectiveness, promoting physiological and psychological recovery of wounded soldiers, preventing chronic pain, and helping them return to the battle field. In recent years, the U.S. Armed Forces have increasingly emphasized prehospital pain management. They have been updating the relevant pain management guidelines to establish a practical system in order to ensure that the pain of wounded soldiers is effectively controlled. This paper reviews the current research on prehospital pain management in the U.S. Armed Forces and summarizes the advanced and practical technologies for prehospital pain management. It provides references for research on prehospital pain management,and optimization of prehospital pain management strategies suited to China’s operational context, and for the design of related training programs.;
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  Reviews

  RFID in blood supply chains: research progress and developments

  LI Penghui, CHEN Yaopeng, YAN Shaoduo

  2025, 49(5): 386-390. https://doi.org/10.7644/j.issn.1674-9960.2025.05.010

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  This paper provides a survey of the research progress in and applications of radio frequency identification（RFID） technology in the entire process management of blood supply. This paper makes an in-depth analysis of the scenarios and effectiveness of the technology,focusing on such key operational stagesas donor management, health screening, blood collection and testing, component preparation, storage and distribution, and patient transfusion. Based on a comprehensive evaluation of current achievements and bottlenecks, this study offers strategies for optimization of RFID technology in the management of blood supply and gives recommendations. The findings are intended to provide data for the establishment of an intelligent and traceable modern blood management system.
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  Research progress in pathogenesis and diagnosis of pulmonary oxygen toxicity

  REN Bohao, MIN Yi, WU Siyuan, WEI Haoyang, HU Jiale, HUANG Guoyang

  2025, 49(5): 391-395. https://doi.org/10.7644/j.issn.1674-9960.2025.05.011

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  High partial pressure oxygen is widely used in the treatment of ischemic and hypoxic diseases and in diving. However, chronic inhalation of gas with high oxygen partial pressure can have a toxic effect on the body, that is, oxygen toxicity. The lung is one of the target organs where injury is the most pronounced and direct after exposure to high partial pressure oxygen. This article reviews the research progress in pathogenesis and diagnosis of pulmonary oxygen toxicity in the hope of providing a reference for related prevention and treatment.
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  Research progress in the molecular mechanism of α2-macroglobulin

  YUAN Fang, JIA Junting, MA Yuyuan

  2025, 49(5): 396-400. https://doi.org/10.7644/j.issn.1674-9960.2025.05.012

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  α 2-Macroglobulin （A2M） is a high-abundance plasma protein with a molecular weight of 720× 10³, containing 1451 residues and 11 domains, and was isolated and identified for the first time in 1946. The capture and inhibitory effect on proteases is the classical biological function of A2M. However, A2M can also interact with membrane receptors, cytokines, and growth factors, and act as a molecular chaperone to affect extracellular protein homeostasis so that it is involved in a wide range of physiological and pathological processes such as immunity, inflammation, and degeneration. This article reviews the structural characteristics of A2M, the reported molecular targets, the mechanism of action, and its biological effects in the hope of providing a new line of thought for the functional exploration and clinical applications of A2M.