1.Assessment of Genotoxicity of Tissue Engineered Materials Based on Improved in vivo Hepatocyte Unscheduled DNA Synthesis(UDS)Assay
Luan-luan WANG ; Li HOU ; Xiang-yu CHU ; Zi-yi YANG ; Ling-xiao SUN ; Xiao-fei WANG ; Qiu-jin QU ; Jing XU ; Zeng-xiang LIU ; Xiao-xia SUN
Progress in Modern Biomedicine 2025;25(17):2740-2748
Objective:An in vivo mammalian hepatocyte Unscheduled DNA Synthesis(UDS)test was used to evaluate the genotoxicity of Cross-linked Sodium Hyaluronate Gel and Bone Repair Materials,providing experimental evidence for establishing a UDS testing method for medical devices and materials.Methods:0.9%sodium chloride injection and cottonseed oil were used as the solvent for test materials and negative control,respectively.N-dimethylnitrosamine(NDMA)was used as the positive control for the early sampling times,and 2-acetylaminofluorene(2-AAF)was used as the positive control for the late sampling times.SD rats were administered a single dose for toxic exposure,and liver tissues were collected at 4 h and 16 h,respectively.Hepatocytes were isolated using collagenase perfusion.After labeling with 5-ethynyl-2'-deoxyuridine(EdU),and the net average fluorescence intensity(NAFI)of cell nuclei and nucleoplasm was measured by fluorescence microscope.Data from 50 cells were used to analyze the DNA repair level.Results:Compared with the negative control groups,the positive control groups(NDMA and 2-AAF)showed highly statistically significant differences in NAFI(P<0.01),indicating successful induction of DNA damage.There was no statistically significant differences between the cross-linked sodium hyaluronate gel groups,bone repair material groups and the negative control group(P>0.05),suggesting that these materials did not significantly induce DNA damage under the experimental conditions.Conclusion:This study first applied EdU labeling technology to the in vivo hepatic UDS assay,achieving non-radioactive labeling through click chemistry reactions.Under the conditions of this study,cross-linked sodium hyaluronate gel and bone repair materials did not exhibit genotoxicity.In the follow-up,the sample range can be expanded and the observation period can be prolonged to further improve the genotoxicity evaluation system of medical devices.
2.Assessment of Genotoxicity of Tissue Engineered Materials Based on Improved in vivo Hepatocyte Unscheduled DNA Synthesis(UDS)Assay
Luan-luan WANG ; Li HOU ; Xiang-yu CHU ; Zi-yi YANG ; Ling-xiao SUN ; Xiao-fei WANG ; Qiu-jin QU ; Jing XU ; Zeng-xiang LIU ; Xiao-xia SUN
Progress in Modern Biomedicine 2025;25(17):2740-2748
Objective:An in vivo mammalian hepatocyte Unscheduled DNA Synthesis(UDS)test was used to evaluate the genotoxicity of Cross-linked Sodium Hyaluronate Gel and Bone Repair Materials,providing experimental evidence for establishing a UDS testing method for medical devices and materials.Methods:0.9%sodium chloride injection and cottonseed oil were used as the solvent for test materials and negative control,respectively.N-dimethylnitrosamine(NDMA)was used as the positive control for the early sampling times,and 2-acetylaminofluorene(2-AAF)was used as the positive control for the late sampling times.SD rats were administered a single dose for toxic exposure,and liver tissues were collected at 4 h and 16 h,respectively.Hepatocytes were isolated using collagenase perfusion.After labeling with 5-ethynyl-2'-deoxyuridine(EdU),and the net average fluorescence intensity(NAFI)of cell nuclei and nucleoplasm was measured by fluorescence microscope.Data from 50 cells were used to analyze the DNA repair level.Results:Compared with the negative control groups,the positive control groups(NDMA and 2-AAF)showed highly statistically significant differences in NAFI(P<0.01),indicating successful induction of DNA damage.There was no statistically significant differences between the cross-linked sodium hyaluronate gel groups,bone repair material groups and the negative control group(P>0.05),suggesting that these materials did not significantly induce DNA damage under the experimental conditions.Conclusion:This study first applied EdU labeling technology to the in vivo hepatic UDS assay,achieving non-radioactive labeling through click chemistry reactions.Under the conditions of this study,cross-linked sodium hyaluronate gel and bone repair materials did not exhibit genotoxicity.In the follow-up,the sample range can be expanded and the observation period can be prolonged to further improve the genotoxicity evaluation system of medical devices.
3.Relationship between depression after ischemic stroke and leukoaraiosis
Jun-Feng QIAN ; Zhi-Jun ZHANG ; Fang-Yuan QIAN ; Ling-Li YANG ; Jun-Xiang LUAN
Chinese Journal of Neuromedicine 2013;12(10):1043-1049
Objective To evaluate the relationship between post-stroke depression (PSD) and leukoaraiosis in a well-defined acute ischemic stroke cohort.Methods One hundred and sixty-eight patients with acute ischemic stroke,admitted to our hospital from June 2007 to March 2011,were examined by magnetic resonance imaging.The patients were made a diagnosis of post-stroke depression by the Diagnostic and Statistical Manual for Mental Disorders,Fourth Edition (DSM-IV) criteria 2 weeks after the index stroke; 60 (35.7%) patients had PSD in the acute period and 60 stroke patients without depression matched according to age and gender served as a control group were chosen.The distribution and severity of leukoaraiosis were evaluated using scale developed by Fazekas and age-related white matter changes (ARWMC) scale.The risk factors of PSD were chosen after univariate analysis and multivariate logistic regression analysis.Results In comparison with the non-PSD group,patients in the PSD group were more likely to have high deep white matter hyperintensities (DWMHs) scores and ARWMC scale scores in the frontal area with significant difference (P<0.05).There were no significant differences in periventricular white matter hypertensities (PVWMHs) scores and ARWMC scale scores in the parieto-occipital area,temporal area,infratentorial area and basal ganglia area between the PSD and control groups (P>0.05).In the multivariate logistic regression analysis,DWMHs remained an independent risk factor of PSD (OR:1.740,95%CI:1.194-2.536,P=0.004).Conclusion There may be an association between PSD and leukoaraiosis; severe leukoaraiosis in the deep area may increase vulnerability to develop depression in the acute period after ischemic stroke.
4.Construction and antigenic evaluation of a recombinant MVA virus-like particle expressing HBV C gene.
Xiang-ling LUAN ; Wei KONG ; Su-jun LIU ; Li LEI ; Yan HU ; Jun HOU ; Hong-hui SHEN ; Yi-chen WU ; Shao-li YOU ; Pan-yong MAO ; Shao-jie XIN
Journal of Southern Medical University 2008;28(2):252-254
OBJECTIVETo construct the virus-like parcel expressing hepatitis B virus (HBV) C gene and identify its immunogenicity.
METHODSHBV C gene was cloned into the shuttle vector pSC11, and the resulted plasmid pSC11-C was transfected into modified vaccinia virus Ankara (MVA).
RESULTSpSC11-C was correctly constructed as verified by sequence analysis and PCR, and the recombinant virus-like parcel possessed good immunogenicity.
CONCLUSIONThe MVA-C expressing HBV C gene has been successfully constructed to provide important basis for gene therapy research of chronic HBV infection.
Genes, Viral ; Genetic Vectors ; Hepatitis B Core Antigens ; genetics ; Recombination, Genetic ; Vaccinia virus ; genetics

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