|
|
|
题名
|
作者
|
年代
|
出处
|
被引量
|
| 1 | miR-193a-3p在食管癌细胞放射抵抗作用的初步研究显示文摘目的:探讨miR-193a-3p在食管鳞癌放射耐受性机制中的作用。方法通过6 MV X射线对4个食管癌细胞系进行照射,采用MTT法检测出相对敏感系及耐受系细胞;茎环引物实时定量PCR法检测miR-193a-3p、miR-155、miR-22-3p在2个细胞中的表达,miR-193a-3p作为表达差异较为明显的 microRNA 被挑选进行下一步研究。分别合成并转染 miR-193a-3p 的 mimic (3PM)或antagomiR (3PA)序列及小干扰RNA (si-LOXL4)以提高或抑制其在细胞中的表达水平,MTT法和流式细胞分析术检测miR-193a-3p及其下游基因LOXL4对于放射敏感性的影响。结果筛选出相对放射敏感细胞系( KYSE510)及耐受细胞系( KYSE410);miR-193a-3p在两系细胞中的表达水平差异显著高于miR-155、miR-22-3p (1.00∶21.13);KYSE510细胞中转染mimic提高其表达后,与对照组相比,其放射敏感性降低,细胞凋亡比例显著下降11.10%(P<0.05),而 KYSE410细胞中转染 antagomiR后,其敏感性增加( P<0.05)。作为miR-193a-3p的下游基因,LOXL4的表达抑制也受到miR-193a-3p的调控,转染si-LOXL4降低其表达,与对照组相比放射敏感性也降低,细胞凋亡比例下降7.07%( P<0.05)。结论 miR-193a-3p可能通过调控基因LOXL4促进食管癌细胞放射耐受性。 | 孟芳 钱立庭 丁伯金 周解平 | 2016 | 中华放射肿瘤学杂志2016，25，8： | 7 |
| 2 | Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles显示文摘The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles(MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease(DNase) and Ribonucleases(RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction(PCR) and reverse transcription PCR(RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria. | Jiuhai Wang Zeeshan Ali Nianyue Wang Wenbiao Liang Hongna Liu Fu Li Haowen Yang Lei He Libo Nie Nongyue He Zhiyang Li | 2015 | Science China Chemistry2015，58，11： | 5 |
| 3 | A结构域阳性纤维连接蛋白促进破骨细胞形成及根尖囊肿骨质破坏显示文摘目的:检测纤维连接蛋白(fibronectin,FN)可变剪接亚型蛋白在根尖囊肿纤维囊壁中的表达,研究每种亚型在体外诱导形成破骨细胞的能力,探讨FN的可变剪接亚型与根尖囊肿病变的进展和骨破坏的关系。方法:选择8例经病理诊断确诊为根尖囊肿患者手术标本,采用免疫组织化学方法,研究每种FN亚型在病变组织中表达强度,分离成纤维细胞,采用反转录-聚合酶链反应(reverse transcription-poymerase chain reaction,RT-PCR)方法检测总FN及其3种亚型(EDA+FN、EDB+FN、CS1-FN)mRNA的表达水平,并采用条件培养基体外诱导破骨细胞分化。根据每种FN亚型的相对表达情况,研究FN亚型与破骨细胞分化之间的关系。结果:在FN产生的亚型中,含蛋白结构域A的纤维连接蛋白(EDA+FN)在纤维囊壁中染色强度高于EDB+FN和CS1-FN。囊壁成纤维细胞中,EDA+FN的mRNA水平显著高于EDB+FN(P=0.007)和CS1-FN(P=0.003),且EDA+FN/总FN值在成纤维细胞的所有亚型中都是最高的(EDB+FN/总FN,P<0.001;CS1-FN/总FN,P<0.001)。体外诱导破骨细胞形成结果显示,EDA+FN/总FN与破骨细胞生成数量呈正相关(R=0.776,P=0.024)。结论:根尖囊肿囊壁成纤维细胞产生各种FN亚型,EDA+FN为主要类型,且EDA+FN/总FN与成纤维细胞诱导破骨细胞效率呈正相关。提示成纤维细胞产生的EDA+FN有利于导致微环境中破骨细胞形成,促进根尖囊肿的骨破坏。 | 陈雅文 王海丞 董伟杰 | 2018 | 口腔医学研究2018，34，9： | 2 |
| 4 | Bevacizumab or fibronectin gene editing inhibits the osteoclastogenic effects of fibroblasts derived from human radicular cysts显示文摘Fibronectin (FN) is a main component of extracellular matrix (ECM) in most adult tissues. Under pathological conditions, particularly inflammation, wound healing and tumors, an alternatively spliced exon extra domain A (EDA) is included in the FN protein (EDA+FN), which facilitates cellular proliferation, motility, and aggressiveness in different lesions. In this study we investigated the effects of EDA+FN on bone destruction in human radicular cysts and explored the possibility of editing FN gene or blocking the related paracrine signaling pathway to inhibit the osteoclastogenesis. The specimens of radicular cysts were obtained from 20 patients. We showed that the vessel density was positively associated with both the lesion size (R=0.49, P=0.001) and EDA+FN staining (R=0.26, P=0.022) in the specimens. We isolated fibroblasts from surgical specimens, and used the CRISPR/Cas system to knockout the EDA exon, or used IST-9 antibody and bevacizumab to block EDA+FN and VEGF, respectively. Compared to control fibroblasts, the fibroblasts from radicular cysts exhibited significantly more Trap+MNCs, the relative expression level of VEGF was positively associated with both the ratio of EDA+FN/total FN (R=0.271, P=0.019) and with the number of Trap+MNCs (R=0.331, P=0.008). The knockout of the EDA exon significantly decreased VEGF expression in the fibroblasts derived from radicular cysts, leading to significantly decreased osteoclastogenesis;similar results were observed using bevacizumab to block VEGF, but block of EDA+FN with IST-9 antibody had no effect. Furthermore, the inhibitory effects of gene editing on Trap+MNC development were restored by exogenous VEGF. These results suggest that EDA+FN facilitates osteoclastogenesis in the fibrous capsule of radicular cysts, through a mechanism mediated by VEGF via an autocrine effect on the fibroblasts. Bevacizumab inhibits osteoclastogenesis in radicular cysts as effectively as the exclusion of the EDA exon by gene editing. | Hai-cheng Wang Peng Wang Yuan-wei Chen Yan Zhang | 2019 | Acta Pharmacologica Sinica2019，40，7： | 0 |
您所在的机构暂未开通此部分的功能，请联系管理员