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目的 观察特异性阻断Hedgehog信号通路对人胰腺癌Mia PaCa-2细胞凋亡的影响,探讨其可能的分子机制.方法 应用特异性Hedgehog通路阻断剂KAAD-eyelopamine阻断Mia PaCa-2细胞的Hedgehog信号通路,噻唑蓝(MTT)法检测阻断Hedgehog信号通路对胰腺癌细胞生长情况的影响;流式细胞技术观察细胞凋亡的变化;Western blot方法检测bax和bcl-2表达的变化.结果 KAAD-cyclopamine明显抑制胰腺癌Mia PaCa-2细胞生长,其作用呈剂量依赖性;阻断Hedgehog信号通路后,胰腺癌细胞凋亡显著增加,细胞中bax表达水平上调,bcl-2表达水平明显下调.结论 特异性阻断Hedgehog信号通路使胰腺癌细胞生长抑制,凋亡明显增加,其机制可能为通过上调bax及下调bcl-2的表达水平实现. 相似文献
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腹腔镜保留脾脏胰体尾切除术适用于胰体尾部良性或低度恶性病变,避免了脾切除术后近、远期并发症,手术方式包括保留脾动静脉的Kimura手术和切除脾动静脉主干、保留胃网膜左血管等侧枝循环的Warshaw手术。腹腔镜下Kimura手术视野清晰,安全可行,术后并发症发生率低,应为保脾胰体尾切除手术的首选。术前检查或术中探查可疑为浸润性恶性病变或病灶与脾血管、脾门关系密切者,应果断放弃保脾术式,改行胰体尾联合脾切除术。 相似文献
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目的 应用慢病毒介导的RNA干扰(RNAi)技术建立稳定低表达胰岛素样生长因子1类受体(IGF1R)基因的人胰腺癌Panc-1细胞株,并观察其对吉西他滨化疗敏感性的影响.方法 设计并合成3条针对IGF1R基因特异性的RNA干扰靶点,构建慢病毒pFU-GW-RNAi载体,经包装后转染293T细胞,获得病毒上清并测定其相应滴度;感染Panc-1细胞,通过实时定量聚合酶链反应(real-time PCR)鉴定RNA干扰效率;筛选出基因沉默效率最高的慢病毒感染人胰腺癌Panc-1细胞,获得稳定转染细胞株.噻唑蓝(MTT)比色法检测干扰IGF1R后Panc-1细胞对吉西他滨敏感性的变化.裸鼠移植瘤模型检测干扰IGF1R前后体内成瘤能力及吉西他滨对移植瘤生长的抑制作用.结果 在Panc-1细胞中成功建立低表达IGF1R基因的细胞株,吉西他滨对Panc-1细胞的半数抑制剂量(IC50)从(0.774±0.001)mg/L下降到(0.330±0.003)mg/L,Panc-1细胞对吉西他滨的敏感性增加了2.35倍.裸鼠移植瘤模型中,干扰IGF1R基因后皮下种植瘤生长缓慢,联合应用吉西他滨后对肿瘤生长的抑制作用进一步增加.结论 慢病毒介导的RNA干扰技术特异性沉默IGF1R基因,能显著增强胰腺癌细胞株Panc-1对吉西他滨的敏感性.Abstract: Objective To construct the lentiviral expression vector for RNA interference of insulin like growth factor-1 receptor (IGF1R) gene in human pancreatic cancer cells and to evaluate its effect on chemosensitivity to gemcitabine.Methods Three sequences of RNA interference targeting IGF1R gene were designed, synthesized and cloned into the pFU-GW-RNAi vector. After transfection of 293T cells with lentiviral vector, the lentivirus was produced and the titer of virus was tested. Panc-1 cells were infected with the lentivirus and the expression of IGF1R mRNA in Panc-1 cells was detected by real-time quantitative polymerase chain reaction (PCR). The changes of gemcitabine sensitivity after RNA interference were examined by methyl thiazol tetrazolium (MTT) assay. In vivo nude mice tumorigenicity model was used to determine the effect of RNA interference targeting IGF1R gene combined with or without gemcitabine on growth inhibition of xenograft tumors.Results A recombinant lentiviral vector expressing shRNA against IGF1R gene was obtained. After RNA interference, the median inhibition concentration (IC50) of gemcitabine against Panc-1 cells was reduced from (0.774±0.001) mg/L to (0.330±0.003) mg/L, indicating that the sensitivity to gemcitabine was increased by 2.35 times after interference. In vivo xenograft formation assay resulted in a significant growth-inhibiting effect by IGF1R RNA interference. Furthermore, the RNA interference targeting IGF1R gene enhanced the antiproliferative effect of gemcitabine on nude mice xenografts.Conclusion The sensitivity of Panc-1 cells to gemcitabine could be enhanced by RNA interference targeting IGF1R gene. 相似文献
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加速康复外科(enhanced recovery after surgery, ERAS)理念及路径正广泛应用于我国临床实践。在此背景下, 中华医学会外科学分会和麻醉学分会共同组织我国数十位普通外科和麻醉科专家, 结合国内外最新文献、我国实际情况以及临床实践经验, 以问题为导向, 以循证为基础, 以基本术式为主要内涵, 历经近1年的筹备、查阅文献、撰写及审校, 于2018年1月在《中国实用外科杂志》及《中华麻醉学杂志》同期发布了《加速康复外科中国专家共识及路径管理指南(2018版)》, 以期在围手术期医学模式下对我国ERAS临床实践的规范化开展起到更好的促进作用。本文对该指南的外科部分进行解读, 诠释其中亮点与热点问题。 相似文献