Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DN...Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray Results Of the 102 rifampin-resistant strains 98 （96.1%） had mutations in the rpoB genes. Conclusion Oligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates.展开更多
Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare w...Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare were used to immunize BALB/c mice.The antigens were evaluated using cellular and humoral immunoassays.The common genes between M.intracellular and M.tuberculosis were identified using genome-wide comparative analysis,and cross-reactive proteins were screened using immunoproteome microarrays.Results Immunization with M.intracellulare proteins induced significantly higher levels of the cytokines interferon-γ(IFN-γ),interleukin-2(IL-2),interleukin-12(IL-12),interleukin-6(IL-6)and immunoglobulins IgG,IgG1,IgM,and IgG2a in mouse serum.Bone marrow-derived macrophages isolated from mice immunized with M.intracellulare antigens displayed significantly lower bacillary loads than those isolated from mice immunized with adjuvants.Whole-genome sequence analysis revealed 396 common genes between M.intracellulare and M.tuberculosis.Microchip hybridization with M.tuberculosis proteins revealed the presence of 478 proteins in the serum of mice immunized with M.intracellulare protein extracts.Sixty common antigens were found using both microchip and genomic comparative analyses.Conclusion This is the advanced study to investigate the immunogenicity of M.intracellulare proteins and the cross-reactive proteins between M.intracellulare and M.tuberculosis.The results revealed the presence of a number of cross-reactive proteins between M.intracellulare and M.tuberculosis.Therefore,this study provides a new way of identifying immunogenic proteins for use in tuberculosis vaccines against both M.intracellulare and M.tuberculosis in future.展开更多
目的探讨甲基转移酶5(methyltransferase-like 5,METTL5)在三阴乳腺癌(triple-negative breast cancer,TNBC)中的作用和潜在机制。方法采用免疫组织化学方法和Western blot检测TNBC肿瘤组织和细胞系中METTL5的表达情况。用靶向METTL5的s...目的探讨甲基转移酶5(methyltransferase-like 5,METTL5)在三阴乳腺癌(triple-negative breast cancer,TNBC)中的作用和潜在机制。方法采用免疫组织化学方法和Western blot检测TNBC肿瘤组织和细胞系中METTL5的表达情况。用靶向METTL5的shRNA(shRNA-METTL5)转染TNBC细胞后,用CCK-8、集落形成、伤口愈合以及Transwell实验分别检测细胞增殖活性、迁移与侵袭,Western blot检测Wnt/β-catenin信号关键蛋白的表达。构建异种移植瘤模型,验证敲降METTL5对TNBC细胞在体内生长以及Wnt/β-catenin信号活性的影响。结果METTL5在TNBC肿瘤组织和细胞系中表达上调(P<0.01)。敲降METTL5可抑制TNBC细胞的增殖、迁移和侵袭并降低了Wnt/β-catenin信号分子β-catenin、细胞周期蛋白(Cyclin)D1、基质金属蛋白酶(MMP)-2和MMP-7的表达(均P<0.01)。体内实验显示,敲降METTL5减缓了移植瘤生长和Wnt/β-catenin信号活性。结论敲降METTL5能抑制TNBC细胞的增殖、迁移与侵袭,其作用可能与抑制Wnt/β-catenin信号通路有关。展开更多
目的:评价基于深度学习的继发性肺结核CT辅助诊断模型在临床应用中的价值。方法:回顾性收集2018年12月至2023年4月在重庆市公共卫生医疗救治中心接受胸部CT平扫的2004例患者的病例资料,分为肺部正常组(544例)、普通肺部感染组(526组)和...目的:评价基于深度学习的继发性肺结核CT辅助诊断模型在临床应用中的价值。方法:回顾性收集2018年12月至2023年4月在重庆市公共卫生医疗救治中心接受胸部CT平扫的2004例患者的病例资料,分为肺部正常组(544例)、普通肺部感染组(526组)和继发性肺结核组(934例)。按照随机分组(通过R语言的sample函数实现训练集和测试集的完全随机分组)的方式,将数据集划分为训练集(1402例,70.0%)和测试集(602例,30.0%)。所有图像采用肺野自动分割算法,获得肺野区域。进一步采用BasicNet和DenseNet算法进行三组间的分类研究。采用曲线下面积(area under curve,AUC)、敏感度、特异度和准确率评价模型的分类性能。最后,在测试数据中,将最优模型与3位不同年资的放射科医生的诊断结果进行比较。结果:602例独立测试集中,DenseNet模型的性能优于BasicNet模型,两种模型的平均AUC、敏感度、特异度和准确率分别为92.1%和89.4%、79.7%和74.0%、89.4%和86.6%、86.2%和83.3%。其中,DenseNet模型的诊断性能优于低年资医生(准确率分别为90.7%和89.1%,Kappa=0.677),与中年资和高年资医生的诊断水平(准确率分别为90.7%、92.2%和95.3%,Kappa值分别为0.746和0.819)保持高度一致性。结论:DenseNet模型能较准确地识别继发性肺结核,与放射科中年资医师的诊断水准相当,可以作为继发性肺结核的辅助诊断工具。展开更多
基金supported by the grant from the National Natural Science Foundation of China (No. 30400018)
文摘Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray Results Of the 102 rifampin-resistant strains 98 （96.1%） had mutations in the rpoB genes. Conclusion Oligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates.
基金supported by National Science and Technology Major Project of China[2018ZX10731301-002]。
文摘Objectives To evaluate the immunogenicity of Mycobacterium intracellulare proteins and determine the cross-reactive proteins between M.intracellulare and M.tuberculosis.Methods Protein extracts from M.intracellulare were used to immunize BALB/c mice.The antigens were evaluated using cellular and humoral immunoassays.The common genes between M.intracellular and M.tuberculosis were identified using genome-wide comparative analysis,and cross-reactive proteins were screened using immunoproteome microarrays.Results Immunization with M.intracellulare proteins induced significantly higher levels of the cytokines interferon-γ(IFN-γ),interleukin-2(IL-2),interleukin-12(IL-12),interleukin-6(IL-6)and immunoglobulins IgG,IgG1,IgM,and IgG2a in mouse serum.Bone marrow-derived macrophages isolated from mice immunized with M.intracellulare antigens displayed significantly lower bacillary loads than those isolated from mice immunized with adjuvants.Whole-genome sequence analysis revealed 396 common genes between M.intracellulare and M.tuberculosis.Microchip hybridization with M.tuberculosis proteins revealed the presence of 478 proteins in the serum of mice immunized with M.intracellulare protein extracts.Sixty common antigens were found using both microchip and genomic comparative analyses.Conclusion This is the advanced study to investigate the immunogenicity of M.intracellulare proteins and the cross-reactive proteins between M.intracellulare and M.tuberculosis.The results revealed the presence of a number of cross-reactive proteins between M.intracellulare and M.tuberculosis.Therefore,this study provides a new way of identifying immunogenic proteins for use in tuberculosis vaccines against both M.intracellulare and M.tuberculosis in future.
文摘目的:评价基于深度学习的继发性肺结核CT辅助诊断模型在临床应用中的价值。方法:回顾性收集2018年12月至2023年4月在重庆市公共卫生医疗救治中心接受胸部CT平扫的2004例患者的病例资料,分为肺部正常组(544例)、普通肺部感染组(526组)和继发性肺结核组(934例)。按照随机分组(通过R语言的sample函数实现训练集和测试集的完全随机分组)的方式,将数据集划分为训练集(1402例,70.0%)和测试集(602例,30.0%)。所有图像采用肺野自动分割算法,获得肺野区域。进一步采用BasicNet和DenseNet算法进行三组间的分类研究。采用曲线下面积(area under curve,AUC)、敏感度、特异度和准确率评价模型的分类性能。最后,在测试数据中,将最优模型与3位不同年资的放射科医生的诊断结果进行比较。结果:602例独立测试集中,DenseNet模型的性能优于BasicNet模型,两种模型的平均AUC、敏感度、特异度和准确率分别为92.1%和89.4%、79.7%和74.0%、89.4%和86.6%、86.2%和83.3%。其中,DenseNet模型的诊断性能优于低年资医生(准确率分别为90.7%和89.1%,Kappa=0.677),与中年资和高年资医生的诊断水平(准确率分别为90.7%、92.2%和95.3%,Kappa值分别为0.746和0.819)保持高度一致性。结论:DenseNet模型能较准确地识别继发性肺结核,与放射科中年资医师的诊断水准相当,可以作为继发性肺结核的辅助诊断工具。
