Background:The role of 1,25-dihydroxyvitamin D3(1,25-(OH)_(2)D_(3))in cancer prevention and treatment is an emerging topic of interest.However,its effects on the stemness of acute myeloid leukemia(AML)cells are poorly...Background:The role of 1,25-dihydroxyvitamin D3(1,25-(OH)_(2)D_(3))in cancer prevention and treatment is an emerging topic of interest.However,its effects on the stemness of acute myeloid leukemia(AML)cells are poorly understood.Methods:The proliferation and differentiation of AML cells(HL60 and NB4)were investigated by the CCK-8 assay,immunocytochemical staining,and flow cytometry.The abilities of HL60 and NB4 cells to form spheres were examined by the cell sphere formation assay.In addition,the levels of stemness-associated markers(SOX2,Nanog,OCT4,and c-Myc)in HL60 and NB4 cells were measured by western blotting and quantitative real-time polymerase chain reaction.Moreover,we obtainedβ-catenin-interacting protein 1(ICAT)-knockout and ICAT-overexpressing HL-60 cells using gene editing and lentiviral infection techniques and investigated the role of ICAT in modulating the stemness-inhibiting effects of 1,25-(OH)_(2)D_(3)using the aforementioned experimental methods.Finally,we validated our findings in vivo using NOD/SCID mice.Results:1,25-(OH)_(2)D_(3)inhibited the proliferation and stemness of AML cells(HL60 and NB4)and induced their differentiation into monocytes.Additionally,the knockdown of ICAT in HL60 cells attenuated the inhibitory effects of 1,25-(OH)_(2)D_(3)on proliferation and stemness and suppressed the expression of stemness markers.Conversely,overexpression of ICAT enhanced the aforementioned inhibitory effects of 1,25-(OH)_(2)D_(3).Consistently,in NOD/SCID mice,1,25-(OH)_(2)D_(3)suppressed tumor formation by HL-60 cells,and the effects of ICAT knockdown or overexpression on 1,25-(OH)_(2)D_(3) aligned with the in vitro findings.Conclusion:1,25-(OH)_(2)D_(3)inhibits AML cell stemness,possibly through modulation of the ICAT-mediated Wnt/β-catenin signaling pathway.展开更多
目的:对HL60细胞在NSC67657作用下向单核系分化前后,双向电泳分离的表达差异蛋白β-catenin相关蛋白1(Beta-catenin-interacting protein 1,ICAT)进行验证,并对ICAT在细胞分化中的功能进行研究.方法:通过RT-PCR和Western blot方法验证...目的:对HL60细胞在NSC67657作用下向单核系分化前后,双向电泳分离的表达差异蛋白β-catenin相关蛋白1(Beta-catenin-interacting protein 1,ICAT)进行验证,并对ICAT在细胞分化中的功能进行研究.方法:通过RT-PCR和Western blot方法验证药物作用细胞前后ICAT基因和蛋白的表达差异;通过免疫荧光协同分析目的蛋白的表达水平,并对其进行初步定位.构建pDsRed-ICAT真核表达载体,转染HL60细胞,筛选阳性克隆.对ICAT基因重组质粒转染细胞作细胞形态学、细胞增殖改变的观察和细胞周期检测以及超微结构观察.结果:NSC67657诱导HL60细胞向单核系分化,ICAT蛋白表达上调,其主要定位于细胞核和胞质.真核表达载体构建成功,电转后G418筛选可得90%以上阳性克隆.转染重组质粒的HL60细胞增殖受抑,电镜下胞核异染色质密集,核质比减小,表面抗原CD14表达和对照组无差异,但在药物处理后24h即可表达71.3%,明显高于对照组,瑞氏染色可见明显分化细胞.结论:ICAT蛋白在NSC67657诱导HL60细胞分化中表达上调,但仅是过表达的ICAT基因并不能诱导HL60细胞向单核系分化,却能提高HL60细胞对NSC67657诱导作用的敏感性.展开更多
AIM: To classify the histological severity of Helicobacter pylori (H. pylori) infection-associated gastritis by confocal laser endomicroscopy (CLE). METHODS: Patients with upper gastrointestinal symptoms or individual...AIM: To classify the histological severity of Helicobacter pylori (H. pylori) infection-associated gastritis by confocal laser endomicroscopy (CLE). METHODS: Patients with upper gastrointestinal symptoms or individuals who were screened for gastric cancer were enrolled in this study. Histological severity of H. pylori infection-associated gastritis was graded according to the established CLE criteria. Diagnostic value of CLE for histo-logical gastritis was investigated and compared with that of white light endoscopy (WLE). Targeted biopsies from the sites observed by CLE were performed. RESULTS: A total of 118 consecutive patients with H. pylori infection-associated gastritis were enrolled in this study. Receiver operating characteristic curve analysis showedthat the sensitivity and specifi city of CLE were 82.9% and 90.9% for the diagnosis of H. pylori infection, 94.6% and 97.4% for predicting gastric normal mucosa, 98.5% and 94.6% for predicting histological active inflammation, 92.9% and 95.2% for predicting glan-dular atrophy, 98.6% and 100% for diagnosing intes-tinal metaplasia, respectively. Post-CLE image analysis showed that goblet cells and absorptive cells were the two most common parameters on the CLE-diagnosed intestinal metaplasia (IM) images (P < 0.001). More his-tological lesions of the stomach could be found by CLE than by WLE (P < 0.001). CONCLUSION: CLE can accurately show the histological severity of H. pylori infection-associated gastritis. Mapping IM by CLE has a rather good diagnostic accuracy.展开更多
We report a unique red light-emitting Eu-doped borosilicate glass to convert color for warm white light-emitting diodes. This glass can be excited from 394 nm-peaked near ultraviolet light, 466 nm-peaked blue light, t...We report a unique red light-emitting Eu-doped borosilicate glass to convert color for warm white light-emitting diodes. This glass can be excited from 394 nm-peaked near ultraviolet light, 466 nm-peaked blue light, to 534 nm- peaked green light to emit the desired red light with an excellent transmission in the wavelength range of 400-700 nm which makes this glass suitable for color conversion without a great cost of luminous power loss. In particular, when assembling this glass for commercial white light-emitting diodes, the tested results show that the color rendering index is improved to 84 with a loss of luminous power by 12 percent at average, making this variety of glass promising for inorganic ＂remote-phosphor＂ color conversion.展开更多
Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related deaths,characterized by highly hypoxic tumor microenvironment.Hypoxia-inducible factor-1α(HIF-1α)is a major regulator involved in cellular ...Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related deaths,characterized by highly hypoxic tumor microenvironment.Hypoxia-inducible factor-1α(HIF-1α)is a major regulator involved in cellular response to changes of oxygen levels,supporting the adaptation of tumor cells to hypoxia.Bruceine D(BD)is an isolated natural quassinoid with multiple anti-cancer effects.Here,we identified BD could significantly inhibit the HIF-1α expression and its subsequently mediated HCC cell metabolism.Using biophysical proteomics approaches,we identified inhibitor of β-catenin and T-cell factor(ICAT)as the functional target of BD.By targeting ICAT,BD disrupted the interaction of bcatenin and ICAT,and promoted β-catenin degradation,which in turn induced the decrease of HIF-1α expression.Furthermore,BD could inhibit HCC cells proliferation and tumor growth in vivo,and knockdown of ICAT substantially increased resistance to BD treatment in vitro.Our data highlight the potential of BD as a modulator of β-catenin/HIF-1α axis mediated HCC metabolism.展开更多
In this study we developed a quantitative proteomic method named ICATswitch by introducing isotope-coded affinity tag(ICAT)reagents into the biotin-switch method,and used it to investigate S-nitrosation in the liver o...In this study we developed a quantitative proteomic method named ICATswitch by introducing isotope-coded affinity tag(ICAT)reagents into the biotin-switch method,and used it to investigate S-nitrosation in the liver of normal control C57BL/6J mice and type 2 diabetic KK-Ay mice.We got fifty-eight S-nitrosated peptides with quantitative information in our research,among which thirty-seven had changed S-nitrosation levels in diabetic mouse liver.The S-nitrosated peptides belonged to fortyeight proteins(twenty-eight were new S-nitrosated proteins),some of which were new targets of S-nitrosation and known to be related with diabetes.S-nitrosation patterns were different between diabetic and normal mice.Gene ontology enrichment results suggested that S-nitrosated proteins are more abundant in amino acid metabolic processes.The network constructed for Snitrosated proteins by text-mining technology provided clues about the relationship between S-nitrosation and type 2 diabetes.Our work provides a new approach for quantifying S-nitrosated proteins and suggests that the integrative functions of S-nitrosation may take part in pathophysiological processes of type 2 diabetes.展开更多
基金supported by the Guangdong Basic and Applied Basic Research Foundation(2022B1515230007).
文摘Background:The role of 1,25-dihydroxyvitamin D3(1,25-(OH)_(2)D_(3))in cancer prevention and treatment is an emerging topic of interest.However,its effects on the stemness of acute myeloid leukemia(AML)cells are poorly understood.Methods:The proliferation and differentiation of AML cells(HL60 and NB4)were investigated by the CCK-8 assay,immunocytochemical staining,and flow cytometry.The abilities of HL60 and NB4 cells to form spheres were examined by the cell sphere formation assay.In addition,the levels of stemness-associated markers(SOX2,Nanog,OCT4,and c-Myc)in HL60 and NB4 cells were measured by western blotting and quantitative real-time polymerase chain reaction.Moreover,we obtainedβ-catenin-interacting protein 1(ICAT)-knockout and ICAT-overexpressing HL-60 cells using gene editing and lentiviral infection techniques and investigated the role of ICAT in modulating the stemness-inhibiting effects of 1,25-(OH)_(2)D_(3)using the aforementioned experimental methods.Finally,we validated our findings in vivo using NOD/SCID mice.Results:1,25-(OH)_(2)D_(3)inhibited the proliferation and stemness of AML cells(HL60 and NB4)and induced their differentiation into monocytes.Additionally,the knockdown of ICAT in HL60 cells attenuated the inhibitory effects of 1,25-(OH)_(2)D_(3)on proliferation and stemness and suppressed the expression of stemness markers.Conversely,overexpression of ICAT enhanced the aforementioned inhibitory effects of 1,25-(OH)_(2)D_(3).Consistently,in NOD/SCID mice,1,25-(OH)_(2)D_(3)suppressed tumor formation by HL-60 cells,and the effects of ICAT knockdown or overexpression on 1,25-(OH)_(2)D_(3) aligned with the in vitro findings.Conclusion:1,25-(OH)_(2)D_(3)inhibits AML cell stemness,possibly through modulation of the ICAT-mediated Wnt/β-catenin signaling pathway.
文摘目的:对HL60细胞在NSC67657作用下向单核系分化前后,双向电泳分离的表达差异蛋白β-catenin相关蛋白1(Beta-catenin-interacting protein 1,ICAT)进行验证,并对ICAT在细胞分化中的功能进行研究.方法:通过RT-PCR和Western blot方法验证药物作用细胞前后ICAT基因和蛋白的表达差异;通过免疫荧光协同分析目的蛋白的表达水平,并对其进行初步定位.构建pDsRed-ICAT真核表达载体,转染HL60细胞,筛选阳性克隆.对ICAT基因重组质粒转染细胞作细胞形态学、细胞增殖改变的观察和细胞周期检测以及超微结构观察.结果:NSC67657诱导HL60细胞向单核系分化,ICAT蛋白表达上调,其主要定位于细胞核和胞质.真核表达载体构建成功,电转后G418筛选可得90%以上阳性克隆.转染重组质粒的HL60细胞增殖受抑,电镜下胞核异染色质密集,核质比减小,表面抗原CD14表达和对照组无差异,但在药物处理后24h即可表达71.3%,明显高于对照组,瑞氏染色可见明显分化细胞.结论:ICAT蛋白在NSC67657诱导HL60细胞分化中表达上调,但仅是过表达的ICAT基因并不能诱导HL60细胞向单核系分化,却能提高HL60细胞对NSC67657诱导作用的敏感性.
基金Supported by A Program from Clinical Projects of Ministry of Health of China (2007) and Taishan Scholar Program of Shandong Province
文摘AIM: To classify the histological severity of Helicobacter pylori (H. pylori) infection-associated gastritis by confocal laser endomicroscopy (CLE). METHODS: Patients with upper gastrointestinal symptoms or individuals who were screened for gastric cancer were enrolled in this study. Histological severity of H. pylori infection-associated gastritis was graded according to the established CLE criteria. Diagnostic value of CLE for histo-logical gastritis was investigated and compared with that of white light endoscopy (WLE). Targeted biopsies from the sites observed by CLE were performed. RESULTS: A total of 118 consecutive patients with H. pylori infection-associated gastritis were enrolled in this study. Receiver operating characteristic curve analysis showedthat the sensitivity and specifi city of CLE were 82.9% and 90.9% for the diagnosis of H. pylori infection, 94.6% and 97.4% for predicting gastric normal mucosa, 98.5% and 94.6% for predicting histological active inflammation, 92.9% and 95.2% for predicting glan-dular atrophy, 98.6% and 100% for diagnosing intes-tinal metaplasia, respectively. Post-CLE image analysis showed that goblet cells and absorptive cells were the two most common parameters on the CLE-diagnosed intestinal metaplasia (IM) images (P < 0.001). More his-tological lesions of the stomach could be found by CLE than by WLE (P < 0.001). CONCLUSION: CLE can accurately show the histological severity of H. pylori infection-associated gastritis. Mapping IM by CLE has a rather good diagnostic accuracy.
基金Project supported by the National Natural Science Foundation of China (Grant Nos. 50872091 and 21076161)the Tianjin Municipal Sci/Tech. Commission, China (Grant Nos. 10SYSYJC28100 and 2006ZD30)the Tianjin Municipal Higher Education Commission, China (Grant No. 20110304)
文摘We report a unique red light-emitting Eu-doped borosilicate glass to convert color for warm white light-emitting diodes. This glass can be excited from 394 nm-peaked near ultraviolet light, 466 nm-peaked blue light, to 534 nm- peaked green light to emit the desired red light with an excellent transmission in the wavelength range of 400-700 nm which makes this glass suitable for color conversion without a great cost of luminous power loss. In particular, when assembling this glass for commercial white light-emitting diodes, the tested results show that the color rendering index is improved to 84 with a loss of luminous power by 12 percent at average, making this variety of glass promising for inorganic ＂remote-phosphor＂ color conversion.
基金supported by the National Natural Science Foundation of China(No.81903654 and 81773941)Program for Professor of Special Appointment(Young Eastern Scholar)at Shanghai Institutions of Higher Learning(QD2018035,China)+4 种基金Shanghai“Chenguang Program”of Education Commission of Shanghai Municipality(No.18CG46,China)Shanghai Sailing Program(No.19YF1449400,China)Ruijin Youth NSFC Cultivation Fund(KY20194297,China)National Key Subject of Drug Innovation(2019ZX09201005-007,China)National Key R&D Program for key research project of modernization of traditional Chinese medicine(2019YFC1711602,China)。
文摘Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related deaths,characterized by highly hypoxic tumor microenvironment.Hypoxia-inducible factor-1α(HIF-1α)is a major regulator involved in cellular response to changes of oxygen levels,supporting the adaptation of tumor cells to hypoxia.Bruceine D(BD)is an isolated natural quassinoid with multiple anti-cancer effects.Here,we identified BD could significantly inhibit the HIF-1α expression and its subsequently mediated HCC cell metabolism.Using biophysical proteomics approaches,we identified inhibitor of β-catenin and T-cell factor(ICAT)as the functional target of BD.By targeting ICAT,BD disrupted the interaction of bcatenin and ICAT,and promoted β-catenin degradation,which in turn induced the decrease of HIF-1α expression.Furthermore,BD could inhibit HCC cells proliferation and tumor growth in vivo,and knockdown of ICAT substantially increased resistance to BD treatment in vitro.Our data highlight the potential of BD as a modulator of β-catenin/HIF-1α axis mediated HCC metabolism.
基金supported by the National Basic Research Program(973 program)of China(Grant Nos.2006CB911001,2005CB522804)the National Natural Science Foundation of China(Grant Nos.90606020 and 30770512)the Knowledge Innovation Program of the Chinese Academy of Sciences.
文摘In this study we developed a quantitative proteomic method named ICATswitch by introducing isotope-coded affinity tag(ICAT)reagents into the biotin-switch method,and used it to investigate S-nitrosation in the liver of normal control C57BL/6J mice and type 2 diabetic KK-Ay mice.We got fifty-eight S-nitrosated peptides with quantitative information in our research,among which thirty-seven had changed S-nitrosation levels in diabetic mouse liver.The S-nitrosated peptides belonged to fortyeight proteins(twenty-eight were new S-nitrosated proteins),some of which were new targets of S-nitrosation and known to be related with diabetes.S-nitrosation patterns were different between diabetic and normal mice.Gene ontology enrichment results suggested that S-nitrosated proteins are more abundant in amino acid metabolic processes.The network constructed for Snitrosated proteins by text-mining technology provided clues about the relationship between S-nitrosation and type 2 diabetes.Our work provides a new approach for quantifying S-nitrosated proteins and suggests that the integrative functions of S-nitrosation may take part in pathophysiological processes of type 2 diabetes.
