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2022 1

2008 1

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内爆炸 1

冲击波 1

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Construction of Hsp90β gene specific silencing plasmid and its transfection efficiency

JI Yewei, NIE Bin, LI Ping, ZHOU Yuanguo, XU Xiaoyu

《医学前沿（英文）》 2007年第1卷第3期页码 253-257 doi: 10.1007/s11684-007-0048-9

摘要： The purpose of this work was to construct the plasmid that could direct the synthesis of siRNA-like transcripts and thus mediate strong and specific repression of human heat shock protein 90β (Hsp90β) gene expression and to compare the transfection efficiency of the plasmids in varying conditions of transfection. Three 64 nt oligos corresponding to different regions of the target gene were chemically synthesized and annealed and were then ligated with pSUPER EGFP1 plasmid and double-digested with HindIII and Π. Recombinant plasmids were transformed into , DH5a, and the colonies were picked and grown in the Amp-agarose. The presence of positive clones was checked by the means of endodigestion and sequencing. Three cell strains, HepG2, Human umbilicus vein endothelium cells (HUVEC) and HeK293, were cultured. Then the plasmids were transfected into the cells at different ratios of plasmid to Lipofectamine. The transfection efficiency was measured by detection of enhanced green fluorescence protein (EGFP). The presence of positive recombinant clones were verified by double-digestion and sequencing. The bases inserted into the plasmids were correct and the positive colonies were named pSuper-Hsp90β1, pSuper-Hsp90β2 and pSuper-Hsp90β3. After optimizing the ratio of plasmid to Lipofectamine, we achieved high transfection efficiency in HeK293 cells. Transfection efficiency was still low in the HepG2 cells. In conclusion, the si-RNA-synthesizing plasmids targeting Hsp90β were constructed and transfected into cells with different transfection efficiency.

关键词： HindIII transfection efficiency detection enhanced different

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Four-protein model for predicting prognostic risk of lung cancer

《医学前沿（英文）》 2022年第16卷第4期页码 618-626 doi: 10.1007/s11684-021-0867-0

摘要： Patients with lung cancer at the same stage may have markedly different overall outcome and a lack of specific biomarker to predict lung cancer outcome. Heat-shock protein 90 β (HSP90β) is overexpressed in various tumor cells. In this study, the ELISA results of HSP90β combined with CEA, CA125, and CYFRA21-1 were used to construct a recursive partitioning decision tree model to establish a four-protein diagnostic model and predict the survival of patients with lung cancer. Survival analysis showed that the recursive partitioning decision tree could distinguish the prognosis between high- and low-risk groups. Results suggested that the joint detection of HSP90β, CEA, CA125, and CYFRA21-1 in the peripheral blood of patients with lung cancer is plausible for early diagnosis and prognosis prediction of lung cancer.

关键词： lung cancer HSP90β decision tree model prognosis

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化爆冲击波在90°拐角通道内的到时规律

沈俊,李永池,庞伟宾,

《中国工程科学》 2006年第8卷第3期页码 53-57

摘要：

对空气冲击波在90°拐角坑道内传播的到时规律及其影响因素进行了实验研究，在无量纲分析基础上建立了简化的工程模型。通过对实验数据拟合建立了可以对高能炸药在90°拐角坑道内爆炸产生的空气冲击波到达时间进行预计的公式，利用该公式可以进一步求得空气冲击波在90°拐角坑道内的传播速度。

关键词：内爆炸冲击波到时

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Prokaryotic expression and purification of fusion protein HSP70-EGFP and its application in the study

QU Ping, SUI Yanfang, MA Jiahai, CHEN Guangsheng, LIU Libing, CHEN Jiankang, LIU Fang'e

《医学前沿（英文）》 2008年第2卷第1期页码 82-86 doi: 10.1007/s11684-008-0015-0

摘要： To study the endocytic activity of dendritic cells (DCs) by obtaining fusion protein HSP70-EGFP as exogenous antigen and loading it with DCs derived from human peripheral blood. Fusion protein HSP70-EGFP was prokaryotically expressed, isolated and purified. DCs were isolated and cultured from human peripheral blood. The DCs were divided into 3 groups in the endocytic experiment. There were 10 DCs in each group. Group 1 and 2 were respectively incubated for 30 min. with HSP70-EGFP and EGFP. Group 3 was incubated with HSP70 for 30 min, and then incubated for 30 min. with HSP70-EGFP. Subsequently, 3 groups were placed in an incubator at 37°C for 0.5, 1, 2 and 24 h. Flow cytometry (FCM) was adopted to detect the amount of DCs with EGFP inside. IL-12 Eli-spot was adopted to detect the amount of DCs which secreted IL-12. There were 5 types in the experiment: LPS, inactive LPS, HSP70-EGFP, EGFP and no antigen. Fusion protein HSP70-EGFP was successfully obtained and its molecular weight was 97 000. It accounted for 35.32% of the total protein. Under irradiation of an ultraviolet lamp, the protein solution sent out viridescent fluorescence. The result detected by FCM indicated that after incubation for 0.5 h at 37°C, the positive rate in group 1 was 63%, while the other 2 groups were negative. After incubation for 1, 2 and 24 h at 37°C, the positive rates in the 3 groups were above 80%. The IL-12 Eli-spot examination shows that with HSP70-EGFP being loaded, the amount of DCs secreting IL-12 was 134.09 ± 31.78/10 cells, a little lower than that of DCs with LPS loaded (with the average point of 156.36 ± 15.73). There was no significant difference between the 2 groups ( < 0.01). By contrast, both of them were significantly higher than inactive LPS-(33.78 ± 1.40)/10 cells and EGFP-loaded (23.13 ± 4.57)/10 cells DC groups in the amount of DCs secreting IL-12 ( < 0.01). The results suggest that receptor-mediated phagocytosis plays a main role in the preliminary stage of DCs internalizing HSP70-EGFP. With increasing incubation time, pinocytosis begins to dominate. HSP70-EGFP may promote DCs to secret cell factor IL-12.

关键词： significant difference irradiation peripheral preliminary EGFP