Objective To investigate the expression profile of microRNA-21 in human cholangiocarcinoma tissues and to validate its bona fide targets in human cholangiocarcinoma cells. Methods The expression profile of microRNA-21...Objective To investigate the expression profile of microRNA-21 in human cholangiocarcinoma tissues and to validate its bona fide targets in human cholangiocarcinoma cells. Methods The expression profile of microRNA-21 in human cholangiocarcinoma tissues and cholangiocarcinoma cell line, QBC939, was evaluated by using real-time PCR analysis. The bona fide targets of microRNA-21 were analyzed and confirmed by dual luciferase reporter gene assay and western blot, respectively. The expressional correlation of microRNA-21 and its targets was probed in human cholangiocarcinoma tissues by using real-time PCR, locked nucleic acid in situ hybridization (LNA-ISH), and immunohistochemistry analysis. Results Real-time PCR analysis revealed that microRNA-21 expression depicted a significant up-regulation in human cholangiocarcinoma tissues about 5.6-fold as compared to the matched normal bileduct tissues (P<0.05). The dual luciferase reporter gene assay revealed endogenous microRNA-21 in cholangiocarcinoma cell line, QBC939, inhibited the luciferase reporter activities of wild-type PTEN (P<0.01) and PDCD4 (P<0.05) and had no this effect on mutated PTEN and PDCD4. Moreover, loss of microRNA-21 function led to a significant increase of PTEN and PDCD4 protein levels in QBC939 cells. Elevated microRNA-21 levels were accompanied by marked reductions of PTEN and PDCD4 expression in the same cholangiocarcinoma tissue. Conclusion microRNA-21 expression is up-regulated in human cholangiocarcinoma and PTEN, PDCD4 are direct effectors of microRNA-21.展开更多
中国正在大力发展氢能,积极实现“3060”碳排放目标,根据测算,中国将成为最大的氢能生产国和消费国之一。中国各省资源禀赋、经济发展情况各不相同,跨省际能源合作尤为重要。为研究绿色氢能跨省供应的可行性,包括氢能运输的技术可行性,...中国正在大力发展氢能,积极实现“3060”碳排放目标,根据测算,中国将成为最大的氢能生产国和消费国之一。中国各省资源禀赋、经济发展情况各不相同,跨省际能源合作尤为重要。为研究绿色氢能跨省供应的可行性,包括氢能运输的技术可行性,采用全成本(total cost of ownership,TCO)理论建立了氢能跨省运输的全成本模型。在此基础上,通过案例研究分析了江西省制氢供粤的相关情况,通过模型计算,得到了江西供粤氢能的成本为31.9元/kg,与当地氢能售价持平。同时还总结出影响外省氢能价格的主要因素,制氢电费占制氢成本的42.38%。研究结果可为中国省际绿色氢能的供给提供解决方案,也可为构建能源运输新通道提供新思路。展开更多
目的:比较二维平铺法和三维悬滴法两种培养体系对小鼠胚胎期生血内皮细胞体外诱导生成造血产物的支持作用。方法:分别利用二维平铺法和三维悬滴法将流式细胞术分选的造血干细胞潜能的生血内皮细胞PK44(CD41-CD43-CD45-CD31^(+)CD201^(+)...目的:比较二维平铺法和三维悬滴法两种培养体系对小鼠胚胎期生血内皮细胞体外诱导生成造血产物的支持作用。方法:分别利用二维平铺法和三维悬滴法将流式细胞术分选的造血干细胞潜能的生血内皮细胞PK44(CD41-CD43-CD45-CD31^(+)CD201^(+)Kit^(+)CD44^(+))群体与OP9-DL1-GFP基质细胞共孵育6 d后,在显微镜下观察两种培养体系产生造血产物的形态特点;利用流式细胞术分析两种培养体系孵育后的造血产物,并统计分析造血产物的比例和绝对数量;分别对两种培养体系共孵育后的造血产物进行造血细胞集落形成单位培养(colony-forming unit culture,CFU-C)实验,并对不同形态的细胞集落进行计数。结果:(1)利用二维平铺法形成的造血细胞平铺或悬浮于基质细胞上,而利用三维悬滴法共孵育后的产物形成了以基质细胞为中心,造血细胞黏附在周围的细胞聚集体。(2)二维平铺法产生更多CD45^(+)造血细胞;三维悬滴法对造血产物的分化具有谱系偏向性,更有利于肥大细胞的分化。(3)二维平铺法诱导生血内皮的分化产物在体外CFU-C实验中可以产生更多的造血集落。结论:二维平铺法更支持小鼠胚内生血内皮细胞体外诱导产生CD45^(+)的造血细胞以及有集落形成能力的造血干祖细胞。展开更多
基金Supported by Institute of Basic Medical Sciences(2009PY13 and 2010PYZ18)the National Natural Science Foundation of China(81100608 and 30901342)
文摘Objective To investigate the expression profile of microRNA-21 in human cholangiocarcinoma tissues and to validate its bona fide targets in human cholangiocarcinoma cells. Methods The expression profile of microRNA-21 in human cholangiocarcinoma tissues and cholangiocarcinoma cell line, QBC939, was evaluated by using real-time PCR analysis. The bona fide targets of microRNA-21 were analyzed and confirmed by dual luciferase reporter gene assay and western blot, respectively. The expressional correlation of microRNA-21 and its targets was probed in human cholangiocarcinoma tissues by using real-time PCR, locked nucleic acid in situ hybridization (LNA-ISH), and immunohistochemistry analysis. Results Real-time PCR analysis revealed that microRNA-21 expression depicted a significant up-regulation in human cholangiocarcinoma tissues about 5.6-fold as compared to the matched normal bileduct tissues (P<0.05). The dual luciferase reporter gene assay revealed endogenous microRNA-21 in cholangiocarcinoma cell line, QBC939, inhibited the luciferase reporter activities of wild-type PTEN (P<0.01) and PDCD4 (P<0.05) and had no this effect on mutated PTEN and PDCD4. Moreover, loss of microRNA-21 function led to a significant increase of PTEN and PDCD4 protein levels in QBC939 cells. Elevated microRNA-21 levels were accompanied by marked reductions of PTEN and PDCD4 expression in the same cholangiocarcinoma tissue. Conclusion microRNA-21 expression is up-regulated in human cholangiocarcinoma and PTEN, PDCD4 are direct effectors of microRNA-21.
文摘中国正在大力发展氢能,积极实现“3060”碳排放目标,根据测算,中国将成为最大的氢能生产国和消费国之一。中国各省资源禀赋、经济发展情况各不相同,跨省际能源合作尤为重要。为研究绿色氢能跨省供应的可行性,包括氢能运输的技术可行性,采用全成本(total cost of ownership,TCO)理论建立了氢能跨省运输的全成本模型。在此基础上,通过案例研究分析了江西省制氢供粤的相关情况,通过模型计算,得到了江西供粤氢能的成本为31.9元/kg,与当地氢能售价持平。同时还总结出影响外省氢能价格的主要因素,制氢电费占制氢成本的42.38%。研究结果可为中国省际绿色氢能的供给提供解决方案,也可为构建能源运输新通道提供新思路。
文摘目的:比较二维平铺法和三维悬滴法两种培养体系对小鼠胚胎期生血内皮细胞体外诱导生成造血产物的支持作用。方法:分别利用二维平铺法和三维悬滴法将流式细胞术分选的造血干细胞潜能的生血内皮细胞PK44(CD41-CD43-CD45-CD31^(+)CD201^(+)Kit^(+)CD44^(+))群体与OP9-DL1-GFP基质细胞共孵育6 d后,在显微镜下观察两种培养体系产生造血产物的形态特点;利用流式细胞术分析两种培养体系孵育后的造血产物,并统计分析造血产物的比例和绝对数量;分别对两种培养体系共孵育后的造血产物进行造血细胞集落形成单位培养(colony-forming unit culture,CFU-C)实验,并对不同形态的细胞集落进行计数。结果:(1)利用二维平铺法形成的造血细胞平铺或悬浮于基质细胞上,而利用三维悬滴法共孵育后的产物形成了以基质细胞为中心,造血细胞黏附在周围的细胞聚集体。(2)二维平铺法产生更多CD45^(+)造血细胞;三维悬滴法对造血产物的分化具有谱系偏向性,更有利于肥大细胞的分化。(3)二维平铺法诱导生血内皮的分化产物在体外CFU-C实验中可以产生更多的造血集落。结论:二维平铺法更支持小鼠胚内生血内皮细胞体外诱导产生CD45^(+)的造血细胞以及有集落形成能力的造血干祖细胞。
