摘要：目的用前列腺素E2刺激结肠癌细胞,观察和探讨可否引起CacyBP/SIP核转位。方法根据hCacyBP/SIP序列设计引物,PCR扩增、酶切、连接、转化等分子克隆法构建含有钙周期素结合蛋白(CacyBP/SIP)的慢病毒重组质粒,PCR鉴定、测序;将构建好的带有绿荧光蛋白的CacyBP-Lentivirus载体转染至结肠癌SW480细胞,用激光共聚焦显微镜检测CacyBP/SIP表达定位;前列腺素E2刺激后激光共聚焦显微镜、Western blot观察CacyBP/SIP在细胞中的定位。结果激光共聚焦和Westernblot发现:前列腺素E2刺激前CacyBP/SIP定位和表达主要在细胞胞质,用100μmol/L前列腺素E2刺激1h,CacyBP/SIP定位和表达在细胞胞质和胞核。结论前列腺素E2刺激可以引起CacyBP/SIP由胞质转位至细胞核。

摘要：目的利用小片段干扰RNA(small interfering RNA,siRNA)抑制S100A6蛋白的表达,为研究CacyBP/SIP核转位机制提供有利的工具。方法设计S100A6的siRNA序列,用脂质体LipofectamineTM 2000将siRNA转染至人结肠癌SW480细胞中,在荧光显微镜下观察转染效率,用Real-time PCR及Western blot方法检测S100A6在人结肠癌SW480细胞内的mRNA和蛋白表达。结果构建了3对S100A6-siRNA,荧光显微镜下检测转染人结肠癌SW480细胞成功;RT-PCR证明了在SW480细胞内S100A6的mRNA表达显著降低(P<0.05);Western blot证明了SW480细胞内S100A6蛋白表达显著降低(P<0.05);与S100A6si-1、S100A6si-3组相比,S100A6si-2组对S100A6的mRNA和蛋白抑制效果最明显(P<0.05)。结论成功构建了S100A6-siRNA,为CacyBP/SIP核转位机制研究提供了有利的工具。

摘要：目的　扩增人核糖体蛋白 L 2 3(RPL 2 3)编码基因序列 ,构建其正、反义真核表达载体 ,分别转染胃癌细胞SGC790 1及胃癌长春新碱耐药细胞 SGC790 1 /VCR,以进一步研究 RPL2 3基因与胃癌多药耐药的关系 .方法　按文献报道的 RPL 2 3核苷酸序列设计合成 PCR引物 ,利用总 RNA抽取试剂盒从人胃癌长春新碱耐药细胞 SGC790 1 /VCR中提取细胞总 RNA,反转录合成 c DNA,再用 PCR方法扩增目的基因序列 .PCR产物经 DNA序列测定证实后 ,通过双酶切 ,将所获目的基因分别按正、反两个方向定向克隆入真核表达载体 pc DNA3.1 (+)中 ,酶切电泳鉴定 .采用脂质体介导的方法将 pc DNA3.1 (+) - RPL 2 3转染 SGC790 1细胞 ,pc DNA3.1(+) - an RPL2 3转染 SGC790 1 /VCR细胞 ,经 G41 8筛选后 ,随机挑选细胞克隆 .通过斑点杂交检测正、反义转染后 RPL2 3m RNA表达水平的变化 .结果　应用 RT- PCR反应扩增获得大小约 0 .5 kb的特异性片段 .经 DNA序列分析 ,证实与文献报道的人 RPL2 3编码区序列一致 .重组正义真核表达载体 pc DNA3.1 (+) - RPL2 3双酶切产生大小约 5.4kb,0 .5kb的片段 ;重组反义真核表达载体 pc DNA3.1 (+) - an RPL2 3双酶切产生大小约 5.4kb,0 .5kb的片段 ,均与预期结果相符 .转染细胞经筛选后 ,随机挑选 。

摘要：目的构建编码人钙周期素结合蛋白(hCacyBP)基因的亚细胞定位载体并进行序列测定。方法 hCacyBP的模板来自人结肠癌细胞株HCT-116的cDNA序列,根据hCacyBP基因序列设计合成引物,采用PCR方法扩增编码hCacyBP的基因片段;将hCacyBP基因定向克隆到亚细胞定位载体pCMV/myc系列:pCMV/myc/nuc(胞核定位载体)、pCMV/myc/cyto(胞质定位载体)。转化*** DH5α感受态细胞,在氨苄青霉素阳性LB培养基平板上筛选出阳性克隆。重组质粒经PCR及双酶切鉴定并进行序列测定。结果从HCT-116的cDNA模板扩增684 bp的hCacyBP基因片段,分别构建重组质粒pCMV/myc/nuc-hCacyBP、pCMV/myc/cyto-hCacyBP,PCR及双酶切鉴定产物大小与预期值相符合,测序分析进一步表明所克隆的基因为编码hCacyBP的基因片段。结论成功构建hCacyBP基因亚细胞定位载体,这将为研究结肠癌胞核、胞质内的hCacyBP功能和研究该基因异常高表达与结肠癌发生发展的关系奠定实验基础。

摘要：传统医学课程由于过于强调本学科的系统性和完整性,使医学基础与临床应用分离。以"器官系统"为中心的整合医学教学模式在国内外广为使用并取得了良好的效果。宁夏医科大学借鉴国内外先进整合教学理念,对临床医学教育进行"消化营养"课程体系改革,积极探索引导学生运用所学知识去解决临床实际问题,实现基础、临床、预防融会贯通的临床思维模式。本文详细介绍了该课程改革的设计、组织、实施及评价情况,探讨整合式课程改革的经验和教训。

摘要：Background: The aim of this study was to determine the value of small-bowel preparation for patients undergoing capsule endoscopy. Methods: The study design was prospective, randomized, and controlled. Eighty patients referred for capsule endoscopy were randomized into two equal groups. Patients in Group A (mean age 54.40 [15.65] years) ingested 2 L of a polyethylene glycol/electrolyte solution 16 hours before the test, whereas patients in Group B (mean age 59.85 [14.58] years) prepared for the procedure by taking only clear liquids during the prior day. The primary outcome evaluated was the effect of bowel preparation on the quality of capsule images, as assessed with an objective scoring system in which cleansing was graded as “adequate”or “inadequate”by blinded examiners. A secondary outcome was the effect of bowel preparation on diagnostic yield. For this purpose, the results of capsule endoscopy were classified as positive findings, findings of uncertain significance, and no findings. Results: Cleansing of the small intestine was considered “adequate" in 36 patients in Group A (90%) vs. 24 patients of Group B (60%) (p = 0.004). A diagnosis was established in 26 patients in Group A (65%) compared with 12 patients in Group B (30%) (p = 0.003). Conclusions: Bowel preparation before capsule endoscopy improves visualization of the small intestine, which may lead to an improvement in diagnostic yield.