1.Visual analysis of the biological effects of tritium discharge from nuclear wastewater using CiteSpace
Ge YAN ; Lin ZHAO ; Jiaying XU ; Juan WANG
Chinese Journal of Radiological Health 2026;35(1):74-82
Objective With the vigorous development of nuclear reactors and controlled thermonuclear fusion research, the release of tritium, the predominant radionuclide in nuclear wastewater, into the environment has attracted widespread attention. Its impact on human health has also become a hot topic of research. This article presents a visual analysis of the literature on the biological effects of tritium ingestion by organisms over the past 70 years, with the aim of elucidating the biological effects of tritiated water and identifying current research hotspots and emerging trends. Methods We retrieved articles on the biological effects of tritium radiation published in the China National Knowledge Infrastructure (CNKI) and Web of Science (WOS) over the past 70 years. CiteSpace software was used to generate visual maps, including annual number of publications, countries of publication, keyword clustering, keyword timeline, keyword burst, and literature co-citation. Results A total of 437 articles were included. The cumulative number of annual publications exhibited a linear growth trend. Research hotspots focused on low-radioactivity tritiated water, dose rate effect, DNA double-strand break damage, genetic effect, and cancer mortality. Emerging research frontiers included human lymphocyte immune injury, oxidase activity, comparison of marine organisms in different living environments, comparison of tritium and ionizing radiation effects, changes in mitochondrial ATP content, and the hormetic effect of low-dose radiation. Conclusion In cellular and animal models, high doses of tritium exposure induce negative biological effects. However, whether low doses of tritium esposure elicit beneficial biological effects remains to be further explored. It is suggested that domestic and foreign teams enhance academic collaboration and discussions, focusing on current hotspots and frontiers to deepen our understanding of the biological effects induced by tritium radiation. This will provide scientific solutions for disease treatment and establish a scientific basis for the safe utilization of nuclear energy and the formulation of safety standards for nuclear wastewater discharge.
2.Mechanisms of Dihuang Yinzi in Treating Advanced Parkinson's Disease Based on Gut Microbiota-SCFAs-inflammation Axis
Renzhi MA ; Yasi LIN ; Tingyue JIANG ; Hongmei ZHU ; Jiayuan LI ; Yu WANG ; Ge ZHANG ; Wenxin FAN ; Jinli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):11-21
ObjectiveTo observe the effects of Dihuang Yinzi (DY) on motor dysfunction in rats with advanced Parkinson's disease (PD) and to investigate the mechanisms by which DY improves advanced PD symptoms through the "gut microbiota-short-chain fatty acids (SCFAs)-inflammation-neuroprotection pathway". MethodsAn advanced PD rat model was induced by rotenone. Rats were divided into a normal group, model group, positive drug group (levodopa, 50 mg·kg-1), and DY low-, medium-, and high-dose groups (5.2, 10.4, 20.8 g·kg-1). After 7 days of administration, motor function was evaluated using the open-field, pole-climbing, and inclined plate tests. Hematoxylin-eosin (HE) staining was used to observe pathological changes in the substantia nigra and colon, and immunohistochemistry was performed to detect α-Synuclein (α-Syn) and tyrosine hydroxylase (TH) expression in the substantia nigra. Enzyme-linked immunosorbent assay (ELISA) was used to measure levels of dopamine (DA), 5-hydroxytryptamine (5-HT), 3,4-dihydroxyphenylacetic acid (DOPAC), Levodopa, homovanillic acid (HVA), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β). Western blot analysis was used to detect the expression of zonula occludens-1 (ZO-1) and occludin. Gut microbiota diversity was analyzed by 16S rRNA sequencing, and gas chromatography (GC) was used to determine the content of SCFAs in colonic contents. ResultsCompared with the normal group, the model group showed significantly decreased movement speed and distance in the open-field test, prolonged pole-climbing time, and reduced retention angle on the inclined plate (P<0.01), accompanied by increased α-Syn expression (P<0.01) and decreased TH expression (P<0.01) in the brain. Compared with the model group, all DY dose groups improved motor dysfunction in advanced PD rats to varying degrees (P<0.05, P<0.01) and alleviated pathological damage in the brain and colon. High-dose DY significantly reduced α-Syn aggregation in the substantia nigra (P<0.01) and increased TH expression (P<0.01). ELISA and Western blot results showed that, compared with the normal group, the model group exhibited decreased levels of DA, 5-HT, DOPAC, Levodopa, and HVA in the striatum (P<0.01), increased levels of TNF-α, IL-6, and IL-1β in the colon and striatum (P<0.01), and significantly reduced expression of ZO-1 (P<0.05) and occludin in the colon (P<0.01). Compared with the model group, all DY dose groups increased the levels of DA, 5-HT, DOPAC, Levodopa, and HVA in the striatum to varying degrees (P<0.05, P<0.01). In the high-dose DY group, the levels of TNF-α, IL-6, and IL-1β in the colon and striatum were reduced (P<0.01), while the expression of ZO-1 (P<0.05) and occludin in the intestine was increased. The 16S rRNA sequencing results indicated that the relative abundances of Actinobacteriota, Enterobacteriaceae, and Erysipelotrichaceae were increased in the model group, whereas the relative abundances of Bacteroidota, class Clostridia, Lachnospiraceae, and Akkermansia muciniphila were decreased. These changes were effectively reversed after high-dose DY intervention. GC analysis showed that the content of SCFAs in the colonic contents of rats in the model group was decreased (P<0.05, P<0.01), while after high-dose DY intervention, the levels of acetate, propionate, isobutyrate, and butyrate were significantly increased (P<0.05, P<0.01). ConclusionDY may exert therapeutic effects in advanced PD by regulating the gut microbiota-SCFAs-inflammation pathway.
3.Genetic analysis and reproductive intervention for 46 Chinese pedigrees affected with Hereditary multiple exostoses.
Lilan SU ; Xiao HU ; Jing DAI ; Zhengxing WAN ; Duo YI ; Shuangfei LI ; Liang HU ; Yueqiu TAN ; Fei GONG ; Ge LIN ; Guangxiu LU ; Qianjun ZHANG ; Juan DU ; Wenbin HE
Chinese Journal of Medical Genetics 2026;43(4):253-258
OBJECTIVE:
To explore the genetic etiology of 46 Chinese pedigrees affected with Hereditary multiple exostoses (HME) and provide genetic counseling and reproductive intervention.
METHODS:
Whole-exome sequencing and Sanger sequencing were carried out on 87 patients from the 46 pedigrees to analyze the variants of EXT1 and EXT2 genes. Pathogenicity of the variants was assessed based on the guidelines from the American College of Medical Genetics and Genomics and Association for Molecular Pathology (ACMG/AMP). Prenatal diagnosis and preimplantation genetic testing (PGT) were provided for couples with identified pathogenic mutations. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: LL-SC-SG-2014-010).
RESULTS:
In total 17 and 22 pathogenic variants were respectively identified in the EXT1 and EXT2 genes, among which 5 EXT1 and 12 EXT2 variants were unreported previously. Three patients with no family history were found to harbor de novo variants of the EXT1 gene. Twenty nine couples had opted for PGT or underwent prenatal diagnosis following natural conception, and 17 healthy babies were born.
CONCLUSION
This study has clarified the genetic etiology of 45 HME pedigrees and identified 17 novel variants, which has enriched the mutational spectrum of the EXT1 and EXT2 genes. Reproductive intervention through PGT and prenatal diagnosis have prevented the recurrence of HME in these families.
Humans
;
Female
;
Male
;
Pedigree
;
Exostoses, Multiple Hereditary/diagnosis*
;
N-Acetylglucosaminyltransferases/genetics*
;
Adult
;
Exostosin 1
;
Asian People/genetics*
;
Genetic Testing
;
Exostosin 2
;
Mutation
;
China
;
Prenatal Diagnosis
;
Pregnancy
;
Genetic Counseling
;
Preimplantation Diagnosis
;
Exome Sequencing
;
East Asian People
4.The role of circadian rhythm and its new strategies for prevention and treatment in oral diseases
WANG Yajun ; ZHANG Lin ; CHENG Chen ; XING Wenmin ; GE Xuejun ; CHENG Fengli ; ZHANG Fang
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(11):986-996
Circadian rhythm is a biological endogenous process regulated by the suprachiasmatic nucleus of the hypothalamus, which transmits light signals to peripheral clocks and synchronizes the body with the external environment through balanced expression of circadian rhythm genes. Working the night shift, sleep disorders, and exposure to artificial light can lead to disturbances in circadian rhythm and genetic imbalances. A substantial body of research has demonstrated that circadian rhythm plays a significant role in the treatment of autoimmune diseases and neurodegenerative disorders, with increasing attention being directed toward their impact on oral health. Disturbances in circadian rhythm primarily affect psycho-neuro-immune mechanisms, oxidative stress responses, and oral microflora through pathways such as the hypothalamic-pituitary-adrenal axis (HPA axis), brain and muscle ARNT-like 1 (BMAL1)-brain-derived neurotrophic factor (BDNF) signaling, and BMAL1-nuclear factor kappa-B (NF-κB) interactions. These disruptions may influence the progression of oral diseases. Certain pharmacological agents (e.g., melatonin, vitamin D, nobiletin, and propofol) have been shown to regulate mood disorders, immune function, and sleep-wake cycles by upregulating BMAL1 expression, thus alleviating disturbances in circadian rhythm. In addition, non-pharmacological interventions, such as sleep management strategies, psychotherapy approaches, and light therapy, also modulate these processes through HPA axis regulation. Currently, the specific mechanisms by which circadian rhythm regulates BDNF levels, T cell subsets, and inflammatory signals—thereby influencing both pathogenesis and treatment outcomes for oral diseases—remain unclear. Future research should focus on elucidating these molecular mechanisms as well as identifying therapeutic targets related to circadian rhythm within the oral health context. Further, multidisciplinary collaboration encompassing pharmacy, sleep behavior studies, and psychology will be instrumental in advancing prevention strategies and treatments for oral diseases.
5.Predicting late aortic complications after acute type A dissection surgery with volumetric measurements in a Singapore cohort.
Jasmine GE ; Vinay Bahadur PANDAY ; Siew-Pang CHAN ; Bernard WEE ; Julian Chi Leung WONG ; Leok Kheng Kristine TEOH ; Moe Thu SAN ; Carlos A MESTRES ; Theodoros KOFIDIS ; Vitaly A SOROKIN
Singapore medical journal 2025;66(9):469-475
INTRODUCTION:
This study was conducted to evaluate the efficacy of postoperative computed tomography (CT) measurements of aortic lumen volumes in predicting aortic-related complications following acute type A aortic dissection (ATAAD) repair.
METHODS:
We conducted a single-institution retrospective aortic volumetric analysis of patients after ascending aorta replacement performed during 2001-2015. The volumetric measurements of total lumen (total-L), true lumen (TL), false lumen (FL), as well as the TL:FL ratio from the first and second postoperative computer angiograms were obtained. A generalised structural equation model was created to analyse the predictive utility of TL:FL ratio.
RESULTS:
One hundred and twenty-five patients underwent surgical intervention, of whom 97 patients were eventually discharged and analysed for postoperative complications. A total of 19 patients were included in the final analysis. Patients with late postoperative aortic complications had a significantly higher FL volume and total-L volume on the first (FL volume P = 0.041, total-L volume P = 0.05) and second (FL volume P = 0.01, total-L volume P = 0.007) postoperative scans. The odds of having aortic complications were raised by 1% with a 1 cm 3 increase in total-L volume and by 2% with a 1 cm 3 increase in FL volume. The TL:FL ratio was significantly lower in patients who developed complications.
CONCLUSION
Postoperative CT volumetric measurements in patients who developed complications are characterised by a significant increase in the FL volume and total-L volume from the first postoperative scans. Patients with disproportionately expanded FL presenting with TL:FL ratios less than 1 were associated with aortic complications. Hence, the TL:FL ratio may be a reliable and useful parameter to monitor postoperative disease progression and to evaluate the risk of late complications in ATAAD patients.
Humans
;
Male
;
Female
;
Retrospective Studies
;
Singapore
;
Aortic Dissection/diagnostic imaging*
;
Middle Aged
;
Postoperative Complications/diagnostic imaging*
;
Aged
;
Tomography, X-Ray Computed
;
Aortic Aneurysm/diagnostic imaging*
;
Aorta/surgery*
;
Adult
;
Treatment Outcome
;
Computed Tomography Angiography
6.Research on super-resolution reconstruction of mass spectrometry imaging using spatially multi-level and self-supervised deep learning network
Chao-long LIN ; Hui YANG ; Ya-hui GE
Chinese Medical Equipment Journal 2025;46(4):1-8
Objective To propose a method for mass spectrometry imaging(MSI)super-resolution reconstruction based on a spatially multi-level and self-supervised deep learning network(SMSDL-Net),aiming to improve the resolution of mass spectrometry images.Methods SMSDL-Net firstly registered histological and mass spectrometry images using a nonlinear transformation.Then a multi-branch Vision Transformer(ViT)was utilized to extract hierarchical features of high-resolution histological images in a self-supervised manner.These features were subsequently combined with the paired low-resolution mass spectrometry data to construct a regression network,which could realize the prediction of high-resolution mass spectrometry information.To validate the performance of the proposed method,the results by the method were compared with those of the traditional bicubic interpolation(BI)methods based on interpolation processing and the deepFERE method based on convolution neural network(CNN)for super-resolution reconstruction of magnesium elemental image of metal mass spectrometry of human liver cancer samples,and the method was also applied to a mouse renal adenocarcinoma metabolite mass spectrometry imaging dataset.Results Compared with the traditional BI methods and the deepFERE multimodal method,the method proposed demonstrated the lowest root mean square error(RMSE=0.015),the highest structural similarity index measure(SSIM=0.84)and the highest R-squared value(R2=0.853)in reconstructing mass spectrometry images.The effectiveness of the method and its potential for precise tissue-specific distinction were validated using the mouse renal adenocarcinoma metabolite MSI dataset.Conclusion Compared with traditional single-modal and pixel-wise regression deep learning methods,the method proposed enhances the quality of high-resolution mass spectrometry image reconstruction and can serve as a novel method for super-resolution reconstruction in the field of mass spectrometry imaging.[Chinese Medical Equipment Journal,2025,46(4):1-8]
7.Predictive efficacy of multimodal MRI-based machine learning models for glioblastoma multiforme MGMT promoter methylation states
Hong-lin LI ; Shi-ting HU ; Zi-heng ZHOU ; Bing LI ; Zhi-ping QI ; Ruo-qi LI ; Kai LIU ; Chun-feng HU ; Hai-tao GE
Chinese Medical Equipment Journal 2025;46(6):7-13
Objective To explore the predictive efficacy of several multimodal MRI-based machine learning models for the promoter methylation states of O6-methylguanine-DNA methyltransferase(MGMT)of glioblastoma muliforme(GBM)patients in terms of the GBM heterogeneity and the complexity of the tumor microenvironment.Methods Firstly,the multimodal MRI images of 317 GBM patients from The University of Pennsylvania Glioblastoma(UPENN-GBM)dataset were pre-processed,with four sequences involved in including T1-weighted imaging(T1WI)sequence,T1-weighted contrast-enhanced imaging(T1CE)sequence,T2-weighted imaging(T2WI)sequence and fluid-attenuated inversion recovery(FLAIR)sequence,and the radiomics features were extracted for two regions of interest(ROIs)such as the tumor core region and the tumor edema region.Secondly,the data of the 317 GBM patients were randomly divided into a training set(254 cases)and a test set(63 cases),which underwent normalization with Z-scores and feature selection and dimensionality reduction with Lasso regression.Finally,three models were established respectively with particle swarm optimization-support vector machine(PSO-SVM),C-support vector classification(C-SVC)and adaptive boosting(adaptive boosting(Adaboost)algorithms,and the predictive efficacy of the three models for glioblastoma multiforme MGMT promoter methylation states were evaluated in terms of accuracy and AUC.Results The Adaboost model based on T2WI sequence and radiomics features of the tumor core region had the highest predictive efficacy with accuracy and AUC values of 67%and 0.74,respectively,higher than those of other combinations of sequences,models and regions of interest.Conclusion The multimodal MRI-based machine learning models can be used for the prediction of glioblastoma multiforme MGMT promoter methylation states,which provides powerful support for personalized treatment and prognostic assessment of GBM.[Chinese Medical Equipment Journal,2025,46(6):7-13]
8.Changing antibiotic resistance profiles of the bacterial strains isolated from geriatric patients in hospitals across China:data from CHINET Antimicrobial Resistance Surveillance Program,2015-2021
Xiaoman AI ; Yunjian HU ; Chunyue GE ; Yang YANG ; Fupin HU ; Demei ZHU ; Yingchun XU ; Xiaojiang ZHANG ; Hui LI ; Ping JI ; Yi XIE ; Mei KANG ; Chuanqing WANG ; Pan FU ; Yuanhong XU ; Ying HUANG ; Ziyong SUN ; Zhongju CHEN ; Yuxing NI ; Jingyong SUN ; Yunzhuo CHU ; Sufei TIAN ; Zhidong HU ; Jin LI ; Yunsong YU ; Jie LIN ; Bin SHAN ; Yan DU ; Sufang GUO ; Lianhua WEI ; Fengmei ZOU ; Hong ZHANG ; Chun WANG ; Chao ZHUO ; Danhong SU ; Dawen GUO ; Jinying ZHAO ; Hua YU ; Xiangning HUANG ; Wen'en LIU ; Yanming LI ; Yan JIN ; Chunhong SHAO ; Xuesong XU ; Chao YAN ; Shanmei WANG ; Yafei CHU ; Lixia ZHANG ; Juan MA ; Shuping ZHOU ; Yan ZHOU ; Lei ZHU ; Jinhua MENG ; Fang DONG ; Zhiyong LÜ ; Fangfang HU ; Han SHEN ; Wanqing ZHOU ; Wei JIA ; Gang LI ; Jinsong WU ; Yuemei LU ; Jihong LI ; Jinju DUAN ; Jianbang KANG ; Xiaobo MA ; Yanping ZHENG ; Ruyi GUO ; Yan ZHU ; Yunsheng CHEN ; Qing MENG ; Shifu WANG ; Xuefei HU ; Jilu SHEN ; Wenhui HUANG ; Ruizhong WANG ; Hua FANG ; Bixia YU ; Yong ZHAO ; Ping GONG ; Kaizhen WENG ; Yirong ZHANG ; Jiangshan LIU ; Longfeng LIAO ; Hongqin GU ; Lin JIANG ; Wen HE ; Shunhong XUE ; Jiao FENG ; Chunlei YUE
Chinese Journal of Infection and Chemotherapy 2025;25(3):290-302
Objective To investigate the antimicrobial resistance of clinical isolates from elderly patients(≥65 years)in major medical institutions across China.Methods Bacterial strains were isolated from elderly patients in 52 hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program during the period from 2015 to 2021.Antimicrobial susceptibility test was carried out by disk diffusion method and automated systems according to the same CHINET protocol.The data were interpreted in accordance with the breakpoints recommended by the Clinical and Laboratory Standards Institute(CLSI)in 2021.Results A total of 514 715 nonduplicate clinical isolates were collected from elderly patients in 52 hospitals from January 1,2015 to December 31,2021.The number of isolates accounted for 34.3%of the total number of clinical isolates from all patients.Overall,21.8%of the 514 715 strains were gram-positive bacteria,and 78.2%were gram-negative bacteria.Majority(90.9%)of the strains were isolated from inpatients.About 42.9%of the strains were isolated from respiratory specimens,and 22.9%were isolated from urine.More than half(60.7%)of the strains were isolated from male patients,and 39.3%isolated from females.About 51.1%of the strains were isolated from patients aged 65-<75 years.The prevalence of methicillin-resistant strains(MRSA)was 38.8%in 32 190 strains of Staphylococcus aureus.No vancomycin-or linezolid-resistant strains were found.The resistance rate of E.faecalis to most antibiotics was significantly lower than that of Enterococcus faecium,but a few vancomycin-resistant strains(0.2%,1.5%)and linezolid-resistant strains(3.4%,0.3%)were found in E.faecalis and E.faecium.The prevalence of penicillin-susceptible S.pneumoniae(PSSP),penicillin-intermediate S.pneumoniae(PISP),and penicillin-resistant S.pneumoniae(PRSP)was 94.3%,4.0%,and 1.7%in nonmeningitis S.pneumoniae isolates.The resistance rates of Klebsiella spp.(Klebsiella pneumoniae 93.2%)to imipenem and meropenem were 20.9%and 22.3%,respectively.Other Enterobacterales species were highly sensitive to carbapenem antibiotics.Only 1.7%-7.8%of other Enterobacterales strains were resistant to carbapenems.The resistance rates of Acinetobacter spp.(Acinetobacter baumannii 90.6%)to imipenem and meropenem were 68.4%and 70.6%respectively,while 28.5%and 24.3%of P.aeruginosa strains were resistant to imipenem and meropenem,respectively.Conclusions The number of clinical isolates from elderly patients is increasing year by year,especially in the 65-<75 age group.Respiratory tract isolates were more prevalent in male elderly patients,and urinary tract isolates were more prevalent in female elderly patients.Klebsiella isolates were increasingly resistant to multiple antimicrobial agents,especially carbapenems.Antimicrobial resistance surveillance is helpful for accurate empirical antimicrobial therapy in elderly patients.
9.Stellate ganglion block alleviates inflammatory response in acute peritonitis rats by regulating α7nAChR expression
Yu'ao QIN ; Xuejun ZOU ; Ge LIU ; Peng YANG ; Lin LI
Chinese Journal of Immunology 2025;41(9):2141-2144
Objective:To observe the effect of stellate ganglion block(SGB)on the inflammatory response in rats with acute peritonitis,and to explore whether the possible anti-inflammatory mechanism related to the activation of α7nAChR receptor-mediated cholinergic anti-inflammatory pathway.Methods:A total of 36 SD rats were randomly divided into three groups,with 12 rats in each group:sham operation group(S group),acute peritonitis group(AP group)and SGB treatment group(SGB group).2%acetic acid(1 ml/100 g)was intraperitoneally injected to prepare acute peritonitis rat model in AP group and SGB group,and corresponding dose of normal saline was injected into S group.Right cervical sympathetic nerve was separated but not ligated in AP group and S group,the right cervical sympathetic nerve was severed and ligated in SGB group.Abdominal aorta blood samples were collected 24 h after opera-tion,and serum levels of IL-1β,IL-6 and TNF-α were measured by ELISA,pathological changes of peritoneal tissue were observed by light microscope and expression of α7nAChR in peritoneal tissue was measured by Western blot.Results:Compared with S group,se-rum levels of IL-1β,IL-6 and TNF-α in AP and SGB groups were significantly increased(P<0.05).Serum levels of IL-1β,IL-6,TNF-α in SGB group were significantly lower than those in AP group(P<0.05).HE staining of peritoneal tissues in each group revealed that the S group maintained intact peritoneal structure with normal cellular arrangement,while the AP group exhibited disorganized archi-tecture with extensive inflammatory cell infiltration,frequent hemorrhage,and prominent necrotic cells.In contrast,the SGB group displayed only mild inflammatory infiltration with preserved cellular morphology and minimal hemorrhagic or necrotic changes.The ex-pression level of α7nAChR protein in SGB group was significantly higher than that in S group and AP group.Conclusion:SGB can re-duce the inflammatory response of acute peritonitis rats,and its mechanism may be related to the cholinergic anti-inflammatory path-way mediated by α7nAChR.
10.Extraction and subculture of neural stem cells from mouse embryonic spinal cord:comparison and analysis on advantages and disadvantages of three commonly used digestive enzymes
Dan LUO ; Zhilin GE ; Yonghui HOU ; Wanshun WANG ; Jiheng ZHAN ; Yu HOU ; Dingkun LIN ; Shudong CHEN
Chinese Journal of Tissue Engineering Research 2025;29(31):6609-6615
BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.


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