目的:双特异性酪氨酸磷酸化调节激酶,也称CDC样激酶(cell division cycle like kinases,CDC-like kinases,CLK),是一种双特异性蛋白激酶,其参与神经系统疾病、代谢异常、肿瘤等多个病理及生理过程,其中CLK3参与肝癌、乳腺癌等多种肿瘤...目的:双特异性酪氨酸磷酸化调节激酶,也称CDC样激酶(cell division cycle like kinases,CDC-like kinases,CLK),是一种双特异性蛋白激酶,其参与神经系统疾病、代谢异常、肿瘤等多个病理及生理过程,其中CLK3参与肝癌、乳腺癌等多种肿瘤的发生和发展,但其在结直肠癌中的表达情况及临床意义尚无相关报道。本研究通过挖掘肿瘤在线数据库,探讨CLK3在结直肠癌中的表达及临床意义。方法:利用基因表达谱交互分析2(Gene Expression Profiling Interactive Analysis 2,GEPIA2)数据库及GEO2R工具对CLK3在肿瘤间及结直肠癌组织和正常组织间作差异分析,获取差异表达结果后利用UALCN(The University of Alabama at Birmingham Cancer Data Analysis Portal)数据库对CLK3表达差异组作Kaplan-Meier生存分析。利用LinkedOmics数据库对CLK3进行共表达分析,并筛选出与之呈正相关及负相关的各50个基因,然后对获取的100个基因进行基因本体(gene ontology,GO)和京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,同时利用TIMER2.0数据库对CLK3进行免疫相关分析。在组织表达层面,利用人类蛋白质图谱(Human Protein Atlas,HPA)数据库查询CLK3在结直肠癌与正常肠道组织的免疫组织化学标本,分析其表达差异。结果:在UALCN数据库中,与正常组织比较,CLK3在结肠癌组织中低表达(P<0.01),而在直肠癌组织中表达差异无统计学意义(P>0.05)。CLK3的表达水平与结直肠癌患者的预后显著相关(P<0.05),表现为其高表达与不良预后相关。GO与KEGG通路富集结果显示:CLK3共表达基因富集于抗原肽呈递、物质代谢合成过程、转录调节过程的酶反应、γ干扰素介导的信号通路、Wnt信号通路及肠道免疫炎症过程等。相关性分析显示:结肠癌中CLK3表达与BRAF、HERS、NTRK1、PIK3CA基因突变相关,直肠癌中CLK3表达与BRAF和PIK3CA基因突变相关。进一步免疫浸润分析结果显示:在结肠癌中,CLK3的表达水平与CD8^(+)T细胞、CD4^(+)T细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05);在直肠癌中,CLK3的表达水平与CD4^(+)T细胞、巨噬细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05)。在免疫组织化学染色切片中,结肠癌和直肠癌CLK3蛋白表达水平均低于正常结肠组织(均P<0.05)。结论:CLK3在结直肠癌中存在差异表达,是结直肠癌生存预后的指标,有望成为结直肠癌的预后评估和临床治疗的有效靶点之一。展开更多
An efficient and scalable electrochemical asymmetric protocol with metal-free catalysts and even without additional oxidants for the cross-dehydrogenative coupling reaction(CDC)of two C(sp^(3))-H bonds is reported.A s...An efficient and scalable electrochemical asymmetric protocol with metal-free catalysts and even without additional oxidants for the cross-dehydrogenative coupling reaction(CDC)of two C(sp^(3))-H bonds is reported.A series of aldehydes including natural products and various substrates containing C(sp^(3))-H bonds including xanthenes,acridines,cycloheptatrienes and even diarylmethane have been shown to undergo asymmetric CDC to afford a series of carbon-carbon bond coupling products with up to 94%yield and 98%ee.Mechanistic studies such as radical clock experiment suggest that the reaction proceeds via nucleophilic attack by enamine under electrochemical conditions.展开更多
Recombinant interleukin-33(IL-33)inhibits tumor growth,but the detailed immunological mechanism is still unknown.IL-33-mediated tumor suppression did not occur in Batf3^(−/−)mice,indicating that conventional type 1 de...Recombinant interleukin-33(IL-33)inhibits tumor growth,but the detailed immunological mechanism is still unknown.IL-33-mediated tumor suppression did not occur in Batf3^(−/−)mice,indicating that conventional type 1 dendritic cells(cDC1s)play a key role in IL-33-mediated antitumor immunity.A population of CD103^(+)cDC1s,which were barely detectable in the spleens of normal mice,increased significantly in the spleens of IL-33-treated mice.The newly emerged splenic CD103^(+)cDC1s were distinct from conventional splenic cDC1s based on their spleen residency,robust effector T-cell priming ability,and surface expression of FCGR3.DCs and DC precursors did not express Suppressor of Tumorigenicity 2(ST2).However,recombinant IL-33 induced spleen-resident FCGR3^(+)CD103^(+)cDC1s,which were found to be differentiated from DC precursors by bystander ST2+immune cells.Through immune cell fractionation and depletion assays,we found that IL-33-primed ST2^(+)basophils play a crucial role in the development of FCGR3^(+)CD103^(+)cDC1s by secreting IL-33-driven extrinsic factors.Recombinant GM-CSF also induced the population of CD103^(+)cDC1s,but the population neither expressed FCGR3 nor induced any discernable antitumor immunity.The population of FCGR3^(+)CD103^(+)cDC1s was also generated in vitro culture of Flt3L-mediated bone marrow-derived DCs(FL-BMDCs)when IL-33 was added in a pre-DC stage of culture.FL-BMDCs generated in the presence of IL-33(FL-33-DCs)offered more potent tumor immunotherapy than control Flt3L-BMDCs(FL-DCs).Human monocyte-derived DCs were also more immunogenic when exposed to IL-33-induced factors.Our findings suggest that recombinant IL-33 or an IL-33-mediated DC vaccine could be an attractive protocol for better tumor immunotherapy.展开更多
文摘目的:双特异性酪氨酸磷酸化调节激酶,也称CDC样激酶(cell division cycle like kinases,CDC-like kinases,CLK),是一种双特异性蛋白激酶,其参与神经系统疾病、代谢异常、肿瘤等多个病理及生理过程,其中CLK3参与肝癌、乳腺癌等多种肿瘤的发生和发展,但其在结直肠癌中的表达情况及临床意义尚无相关报道。本研究通过挖掘肿瘤在线数据库,探讨CLK3在结直肠癌中的表达及临床意义。方法:利用基因表达谱交互分析2(Gene Expression Profiling Interactive Analysis 2,GEPIA2)数据库及GEO2R工具对CLK3在肿瘤间及结直肠癌组织和正常组织间作差异分析,获取差异表达结果后利用UALCN(The University of Alabama at Birmingham Cancer Data Analysis Portal)数据库对CLK3表达差异组作Kaplan-Meier生存分析。利用LinkedOmics数据库对CLK3进行共表达分析,并筛选出与之呈正相关及负相关的各50个基因,然后对获取的100个基因进行基因本体(gene ontology,GO)和京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,同时利用TIMER2.0数据库对CLK3进行免疫相关分析。在组织表达层面,利用人类蛋白质图谱(Human Protein Atlas,HPA)数据库查询CLK3在结直肠癌与正常肠道组织的免疫组织化学标本,分析其表达差异。结果:在UALCN数据库中,与正常组织比较,CLK3在结肠癌组织中低表达(P<0.01),而在直肠癌组织中表达差异无统计学意义(P>0.05)。CLK3的表达水平与结直肠癌患者的预后显著相关(P<0.05),表现为其高表达与不良预后相关。GO与KEGG通路富集结果显示:CLK3共表达基因富集于抗原肽呈递、物质代谢合成过程、转录调节过程的酶反应、γ干扰素介导的信号通路、Wnt信号通路及肠道免疫炎症过程等。相关性分析显示:结肠癌中CLK3表达与BRAF、HERS、NTRK1、PIK3CA基因突变相关,直肠癌中CLK3表达与BRAF和PIK3CA基因突变相关。进一步免疫浸润分析结果显示:在结肠癌中,CLK3的表达水平与CD8^(+)T细胞、CD4^(+)T细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05);在直肠癌中,CLK3的表达水平与CD4^(+)T细胞、巨噬细胞、中性粒细胞和树突状细胞的浸润水平呈正相关(均P<0.05)。在免疫组织化学染色切片中,结肠癌和直肠癌CLK3蛋白表达水平均低于正常结肠组织(均P<0.05)。结论:CLK3在结直肠癌中存在差异表达,是结直肠癌生存预后的指标,有望成为结直肠癌的预后评估和临床治疗的有效靶点之一。
基金National Natural Science Foundation of China(Nos.22161008,22061003)Guangxi Science and Technology Base and Talent Project(High Level Innovative Talents and Team Training)(Guike No.AD23026094)Guangxi Natural Science Foundation of China(No.2021GXNSFFA220005)for financial support。
文摘An efficient and scalable electrochemical asymmetric protocol with metal-free catalysts and even without additional oxidants for the cross-dehydrogenative coupling reaction(CDC)of two C(sp^(3))-H bonds is reported.A series of aldehydes including natural products and various substrates containing C(sp^(3))-H bonds including xanthenes,acridines,cycloheptatrienes and even diarylmethane have been shown to undergo asymmetric CDC to afford a series of carbon-carbon bond coupling products with up to 94%yield and 98%ee.Mechanistic studies such as radical clock experiment suggest that the reaction proceeds via nucleophilic attack by enamine under electrochemical conditions.
基金the National Research Foundation of Korea(SRC-2017R1A5A1014560). This work was supported by grants from the National Research Foundation of Korea(SRC-2017R1A5A1014560)。
文摘Recombinant interleukin-33(IL-33)inhibits tumor growth,but the detailed immunological mechanism is still unknown.IL-33-mediated tumor suppression did not occur in Batf3^(−/−)mice,indicating that conventional type 1 dendritic cells(cDC1s)play a key role in IL-33-mediated antitumor immunity.A population of CD103^(+)cDC1s,which were barely detectable in the spleens of normal mice,increased significantly in the spleens of IL-33-treated mice.The newly emerged splenic CD103^(+)cDC1s were distinct from conventional splenic cDC1s based on their spleen residency,robust effector T-cell priming ability,and surface expression of FCGR3.DCs and DC precursors did not express Suppressor of Tumorigenicity 2(ST2).However,recombinant IL-33 induced spleen-resident FCGR3^(+)CD103^(+)cDC1s,which were found to be differentiated from DC precursors by bystander ST2+immune cells.Through immune cell fractionation and depletion assays,we found that IL-33-primed ST2^(+)basophils play a crucial role in the development of FCGR3^(+)CD103^(+)cDC1s by secreting IL-33-driven extrinsic factors.Recombinant GM-CSF also induced the population of CD103^(+)cDC1s,but the population neither expressed FCGR3 nor induced any discernable antitumor immunity.The population of FCGR3^(+)CD103^(+)cDC1s was also generated in vitro culture of Flt3L-mediated bone marrow-derived DCs(FL-BMDCs)when IL-33 was added in a pre-DC stage of culture.FL-BMDCs generated in the presence of IL-33(FL-33-DCs)offered more potent tumor immunotherapy than control Flt3L-BMDCs(FL-DCs).Human monocyte-derived DCs were also more immunogenic when exposed to IL-33-induced factors.Our findings suggest that recombinant IL-33 or an IL-33-mediated DC vaccine could be an attractive protocol for better tumor immunotherapy.
