摘要：目的构建靶向存活素(survivin)基因的短发夹干扰RNA(shRNA)表达载体,导入人大肠癌细胞SW480中,研究靶向抑制survivin基因对SW480细胞侵袭和转移的影响。方法根据siRNA设计原则,在survivin序列中选取设计含19个核苷酸(19nt)的靶序列,间以9个核苷酸的茎环序列,两端分别加上对应的酶切位点,形成shRNA的DNA模板并克隆到shRNA表达载体pRNAT-U6.1/Neo中,获得靶向抑制survivin基因的sihNA表达载体pRNAT-U6.1/Neo-survivin;采用Lipofectamine2000TM转染试剂将干扰质粒pRNAT-U6.1/Neo-survivin导入到大肠癌细胞SW480中;用Westernblotting从蛋白水平检测干扰效果;分别采用肿瘤侵袭粘附实验和明胶酶谱分析法检测pRNAT-U6.1/Neo-survivin对SW480细胞的侵袭和转移潜能的影响。结果Survivin基因的蛋白表达均得到显著抑制;沉默SW480的survivin基因可以显著抑制SW480细胞的侵袭和转移潜能,而且survivin基因表达被抑制后,SW480细胞分泌基质金属蛋白酶明显减少。结论Survivin基因可能在SW480的侵袭和转移潜能中起重要作用,沉默SW480的survivin基因可以显著抑制其侵袭、转移和基质金属蛋白酶分泌,因而survivin影响SW480的侵袭和转移可能与调控基质金属蛋白酶分泌密切相关。

摘要：目的克隆migfilin-N末端基因并使其在大肠杆菌中高效表达,纯化基因表达产物并进行多克隆抗体制备,为研究migfilin与大肠癌的关系奠定基础。方法根据人migfilin全长cDNA系列,设计PCR引物,以人大肠癌cDNA文库为模板克隆migfilin-N末端基因,将扩增产物克隆至大肠杆菌表达载体pGEX-4T-1中,经EcoRI/XhoI双酶切鉴定后,进行DNA序列测定。GST-migfilin-N融合蛋白在硫代半乳糖苷诱导下在大肠杆菌BL21中得到表达。利用谷胱甘肽亲和层析纯化融合蛋白,用纯化的GST-migfilin-N融合蛋白免疫家兔制备多克隆抗体。并用纯化的抗migfilin多克隆抗体在不同的细胞株中用Western blotting检测migfilin的表达。结果成功克隆了migfilin-N端基因片段,表达并纯化了GST-migfilin-N融合蛋白,制备了migfilin特异性抗体。Western blotting检测migfilin在不同细胞株中的表达,结果显示migfilin在肺癌及大肠癌细胞株中高表达。结论migfilin特异性抗体制备成功,为深入探讨migfilin在大肠癌中的作用奠定了基础。

摘要：The present study aimed at determining whether short-term exposure to art in shared common areas in congregate housing units could affect health and health determinants among the residents. Ten residents (mean age 80.4 years) at one block were exposed to visual art environmental enrichment in common areas over a period of three months. Thirteen persons (mean age 86.6 years) living in another block played in-house boule. Cornell’s test, Mini-Mental tests (MMT), and face recognition test were performed to assess depression, cognition, and episodic memory, respectively before and after the intervention. The results show that visual art environmental enrichment in common areas and lack of stimulating and guiding dialogues show a change in depression scores in the intervention group (p = 0.018) and the control group (p = 0.009). MMT scores improved only in the control group (p = 0.003). No changes in episodic memory in any of the groups were observed. It could be concluded that in order to obtain a positive result of short term visual art environmental enrichment, guiding art dialogues conducted by nurses, as described in previous research, should be added to visual art environmental enrichment in healthcare settings.

摘要：目的克隆PAK5-N端基因并诱导其表达,进行多克隆抗体制备,为研究其在牙胚细胞中的生物学功能奠定基础。方法根据人全长PAK5cDNA序列,设计引物;利用PCR技术,以PAK5全长cDNA为模板扩增PAK5-N端基因,将扩增产物克隆至pGEX-4T-1载体中,经EcoRI/XhoI双酶切后进行重组质粒鉴定,DNA测序。以硫代半乳糖苷诱导其在大肠杆菌BL21中表达。利用GST融合蛋白纯化系统进行蛋白纯化,通过免疫家兔制备多克隆抗体,并在牙胚细胞中进行了初步研究。结果和结论成功克隆了PAK5-N端基因,在***中表达了PAK5-NT,并纯化了GST融合蛋白,制备了PAK5特异性抗体。Westernblotting表明PAK5在牙胚细胞中过度表达,为其在口腔细胞中的生物学功能研究提供了依据。