不同区域晶状体上皮细胞基因表达差异分析

【目的】比较不同区域猪晶状体上皮细胞的基因表达在转录组水平上的差异。【方法】解剖显微镜下分离晶状体前囊膜，将附着于其上的上皮细胞分为中央（直径为10．56mm）和周边两部分。分别提取两个样本的总RNAs并经PCR扩增，以Cy3和Cy5分别标记扩增的中央与周边部分的cDNA。与含7548个基因的表达谱芯片杂交，经图像分析，生物信息学处理获得基因表达在转录水平差异的相关信息。【结果】中央与周边区域的猪晶状体上皮细胞在转录组水平共鉴定出952个有效表达的基因点，其中差异表达基因261个．以中央区域为参照，周边上皮细胞mRNA上调137个，下调124个。差异表达基因主要涉及的功能有：细胞周期与凋亡、细胞骨架蛋白及细胞外基质、转录、细胞信号分子等。【结论】中央与周边区域猪晶状体上皮细胞基因表达在转录组水平上差异明显。这类差异呈明显的功能聚类。     

A trip through the signaling pathways of melanoma

Many cellular signaling pathways are involved in the development of cancer. Depending on the tumor entity, the nature as well as the mode of activation can differ. Some signaling pathways frequently show changes as all tumor cells have to fulfill some basic requirements such as independence from growth factors or insensitivity against apoptosis. In this review, the possibilities of a tumor to manipulate signaling pathways to reach these goals are exemplified based on an archetypical melanoma cell. In addition, new therapeutic options based on the knowledge of signaling pathways will be discussed.     

Ex Vivo Application of Carbon Monoxide in University of Wisconsin Solution to Prevent Intestinal Cold Ischemia/Reperfusion Injury

Carbon monoxide (CO), a byproduct of heme catalysis, was shown to have potent cytoprotective and anti-inflammatory effects. In vivo recipient CO inhalation at low concentrations prevented ischemia/reperfusion (I/R) injury associated with small intestinal transplantation (SITx). This study examined whether ex vivo delivery of CO in University of Wisconsin (UW) solution could ameliorate intestinal I/R injury. Orthotopic syngenic SITx was performed in Lewis rats after 6 h cold preservation in control UW or UW that was bubbled with CO gas (0.1-5%) (CO-UW). Recipient survival with intestinal grafts preserved in 5%, but not 0.1%, CO-UW improved to 86.7% (13/15) from 53% (9/17) with control UW. At 3 h after SITx, grafts stored in 5% CO-UW showed improved intestinal barrier function, less mucosal denudation and reduced inflammatory mediator upregulation compared to those in control UW. Preservation in CO-UW associated with reduced vascular resistance (end preservation), increased graft cyclic guanosine monophosphate levels (1 h), and improved graft blood flow (1 h). Protective effects of CO-UW were reversed by ODQ, an inhibitor of soluble guanylyl cyclase. In vitro culture experiment also showed better preservation of vascular endothelial cells with CO-UW. The study suggests that ex vivo CO delivery into UW solution would be a simple and innovative therapeutic strategy to prevent transplant-induced I/R injury.     

人精子体外氧化损伤对线粒体tRNA~(LeuUUR)基因的影响

目的:探讨活性氧(ROS)对人类精子线粒体tRNALeuUUR基因的氧化损伤。方法:采用Percoll梯度离心法筛选具有正常生理功能的精子,作为正常精子模型,并分为损伤组20例和对照组20例,分别加入次黄嘌呤-黄嘌呤氧化酶体系或不予处理,37℃有氧环境中孵育60min。分别提取精子DNA,以Fpg酶切损伤碱基并采用接头介导PCR(LM-PCR)检测线粒体tRNALeuUUR基因的氧化损伤。采用Rhodamine(Rh123)荧光探针标记精子,通过流式细胞仪检测线粒体膜电位,观察精子的功能。结果:与对照组相比,损伤组精子孵育后线粒体膜电位明显降低[(116.27±11.72)%vs(64.00±4.88)%,P<0.05]。Fpg酶切和LM-PCR显示精子线粒体tRNALeuUUR基因损伤。结论:ROS可能通过对精子线粒体tRNALeuUUR基因氧化损伤而影响精子功能(线粒体膜电位明显降低),从而引起不育。     

Survivin在食管癌中的表达及其与bcl-2表达关系的相关研究

目的探讨Survivin在食管癌组织中的表达及其与bcl-2蛋白表达的相关性。方法应用免疫组织化学SP法,检测Survivin、bcl-2蛋白在68例食管癌组织和20例正常食管组织中的表达。结果Survivin蛋白在正常食管组织中低表达或不表达,68例食管癌组织中,49例表达阳性,占72.1%。Survivin蛋白表达与分化程度、淋巴结转移密切相关(P<0.05)。食管癌组织bcl-2蛋白表达阳性、阴性组中,Survivin蛋白阳性表达率分别为94.7%(36/38)和43.3%(13/30),两者比较,差异有统计学意义(P<0.05)。Survivin蛋白表达与bcl-2蛋白表达密切相关。结论Survivin在食管癌组织中表达上调,通过抑制细胞凋亡,在食管癌的发生中起到重要作用;凋亡相关基因bcl-2的上调与Survivin的表达可能在食管癌变中起协同作用。     

PTEN及Bcl-2蛋白在膀胱移行细胞癌中的表达及其意义

目的：探讨PTEN及Bcl-2蛋白在膀胱移行细胞癌的表达规律及其临床意义。方法：应用免疫组织化学链亲和素-生物素-霉复合物（SP）方法检测43例膀胱移行细胞癌和7例正常黏膜组织中PTEN及Bcl-2蛋白的表达．分析二者的表达与膀胱癌病理参数的关系。结果：（1）G1、G2、G3肿瘤PTEN表达阳性率分别为85．7％、80．4％、73．3％，浸润性肿瘤和表浅性肿瘤表达阳性率分别为70．4％和93．8％，说明PTEN表达阳性率与肿瘤病理分级、临床分期有关。（2）G。、G2、G3肿瘤Bcl-2表达阳性率分别为14．2％、57．14％、66．67％，浸润性肿瘤和表浅性肿瘤表达阳性率分别为59．3％和43．8％，说明Bcl-2表达阳性率与肿瘤病理分级、临床分期有关。（3）随着肿瘤恶性程度的增高和临床分期进展，PTEN蛋白阳性率呈下降，Bcl-2表达呈增高趋势，二者表达呈负相关关系。结论：PTEN的抑癌作用可能与Bcl-2有关．二者的异常表达在膀胱移行细胞癌的发生、发展过程中起重要作用。二者同时检测有助于判断预后。