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靶向人GSK-3β基因的shRNA真核表达质粒的构建及其在人胰腺癌细胞株PANC-1中的稳定表达

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华中科技大学学报（医学版） 2011年第2期40卷 142-146页

作者：汪理勾善淼周伟王统林刘涛王春友华中科技大学同济医学院附属协和医院普通外科武汉430022

目的构建针对人糖原合成激酶-3β(GSK-3β)基因的shRNA真核表达质粒,并筛选基因沉默效果最明显的shRNA质粒表达载体;转染人胰腺癌细胞株PANC-1,建立稳定表达GSK-3β shRNA的细胞模型。方法针对GSK-3β基因的mRNA序列设计并分别构建3个sh... 详细信息

目的构建针对人糖原合成激酶-3β(GSK-3β)基因的shRNA真核表达质粒,并筛选基因沉默效果最明显的shRNA质粒表达载体;转染人胰腺癌细胞株PANC-1,建立稳定表达GSK-3β shRNA的细胞模型。方法针对GSK-3β基因的mRNA序列设计并分别构建3个shRNA质粒表达载体和1个阴性对照质粒表达载体,经大肠埃希菌扩增,酶切、PCR、测序鉴定,转染胰腺癌PANC-1细胞,采用Real-time PCR检测GSK-3β mRNA被抑制情况。选取抑制效应最强的重组质粒和阴性质粒转染的PANC-1细胞,经G418筛选后,建立稳定表达GSK-3-β shRNA的PANC-1细胞株(实验组)和稳定表达control-shRNA的PANC-1细胞株(阴性对照组)。分别采用荧光显微镜和流式细胞术观察细胞的转染情况;Real-time PCR和Western blot分析GSK-3β的表达。结果①经测序证实,成功构建GSK-3-β shRNA真核表达质粒,插入的DNA片段的序列与设计序列完全一致;②重组质粒瞬时转染PANC-1细胞后,GSK-3β mRNA明显下调(P2号重组质粒效应最强;③重组质粒稳定转染后检测PANC-1细胞中GSK-3β的表达,Real-time PCR和Western blot结果均提示:与未转染的阴性对照组和载体对照组比较,实验组中GSK-3β mRNA和蛋白的表达均显著降低(均P<0.05)。结论成功构建了携带以GSK-3β为靶向的shRNA的重组质粒。经脂质体途径稳定转染PANC-1细胞,该shRNA能够显著抑制GSK-3β的表达。

关键词：胰腺癌糖原合成激酶-3β 转染 RNA干扰短发夹状RNA

GLOBAL ATTRACTIVITY OF POPULATION MODELS WITH DELAYS AND DIFFUSION

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Journal of Systems science & Complexity 2005年第3期18卷 396-403页

作者： QIU Zhipeng Department of Applied Mathematics, Nanjing university of science and technology, Nanjing 210094 Department of Mathematics, university of science and technology of china, Hefei 230026, china. Department of Applied Mathematics Nanjing University of Science and Technology 江苏 Nanjing 210094

In this paper, the asymptotic behavior of three types of population models with delays and diffusion is studied. The first represents one species growth in the patch Ω and periodic environment and with delays recruitment, the second models a single species dispersal among the m patches of a heterogeneous environment, and the third models the spread of bacterial infections. Sufficient conditions for the global attractivity of periodic solution are obtained by the method of monotone theory and strongly concave operators. Some earlier results are extended to population models with delays and diffusion.

关键词： delays population models global attractivity reaction-diffusion

THERMODYNAMIC FORMALISM FOR SUBADDITIVE SEQUENCE OF DISCONTINUOUS FUNCTIONS

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Analysis in Theory and Applications 2010年第2期26卷 174-185页

作者： Xianfeng Ma (Nanjing Normal university/East china university of science and technology, china) Ercai Chen (Nanjing Normal university/Nanjing university, china) Yu Pei (Nanjing Normal university, china) School of Mathematics and Computer science Nanjing Normal Uniersity Nanjing P. R. China Department of Mathematics East China University of Science and Technology Shanghai P. R. China Center of Nonlinear science Nanjing University Nanjing P. R. China

The topological pressure for subadditive sequence of discontinuous functions is defined on any invariant subset having a nested family of subsets in the compact metric space. Two subadditive variational principles associated with two different relatively weak conditions are developed for the defined topological pressure. As an application, we give an example on systems with nonzero Lyapunov exponents.

关键词： thermodynamic formalism variational principle topological pressure subadditive sequence of functions

Differentially Expression of Tual,a Tubulin-encoding Gene,during Flowering of Tea Plant Camellia sinensis (L.) *** Using cDNA Amplified Fragment Length Polymorphism Technique

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Acta Biochimica et Biophysica Sinica 2006年第9期38卷 653-662页

作者： Wan-Ping FANG~(1,2) Chang-Jun JIANG~1* Mei YU~1 Ai-Hua YE~1 Zhao-Xia WANG~(1,3) 1 Key Laboratory of Tea Biochemistry and Biotechnology,Ministry of Agriculture,Anhui Agricultural university,Hefei 230036,china 2 college Of Horticulture,Nanjing Agriculture university,Nanjing 210095,china 3 Auhui Institute of Education,Hefei 230036,china Key Laboratory of Tea Biochemistry and Biotechnology Ministry of Agriculture Anhui Agricultural UniversityHefei 230036 China college Of Horticulture Nanjing Agriculture UniversityNanjing 210095 China Auhui Institute of EducationHefei 230036 China

The complementary DNA （cDNA） amplified fragment length polymorphism technique wasused to isolate transcript-derived fragments corresponding to genes involved in the flowering of tea *** sequence analysis of an approximately 300 bp differential fragment amplified by primer combi-nation E11M11revealed 80%-84% similarity to the corresponding part of an α-tubulin gene of other *** complete cDNA sequence of this α-tubulin was cloned by the rapid amplification of cDNA ends technique;its full length is 1537 bp and contains an open reading frame of 450 amino acid residues with two N-glycosylation sites and four protein kinase C phosphorylation *** deduced amino acid sequences didshow significant homology to the α-tubulin from other plants that has been reported to be a pollen-specificprotein and could be correlated with plant cytoplasm-nucleus-interacted male *** named this completecDNA *** nucleotide and amino acid sequence data of Tual have been recorded in the GenBanksequence database with the accession *** Tual gene was cloned into the pET-32aexpression system and expressed in Escherichia coli BL21trxB（DE3）.The molecular weight of expressedprotein was deduced to be approximately 49 *** blot analysis was used to identify the temporalexpression of Tual in tea *** studies of the effect of Tual protein on pollen tube growth indicatedthe Tual solution obviously promoted the growth of tea pollen tube.

关键词： cDNA-AFLP Camellia sinensis flower bud α-tubulin Western blot pollen

Quality Control of Postoperative Acute Pain Service

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Journal of huazhong university of science and technology(medical sciences) 1999年第4期19卷 310-313页

作者：张小铭吕阳胡晓敏姚尚龙曾邦雄 Department of Anesthesia Xiehe Hospital Tongji Medical University Wuhan

To establish an effective method of continuous quality control of acute pain service, a retrospective study on incident reporting during postoperative anal-gesia period was conducted. Incidents were reported and analyzed in 1507 patients who received epidural postoperative analgesia, and the results of satisfaction of pain relief was compared with those of incident analysis. In this study, an incident was defined as any factor that might or had affected patient's safety during analgesia period. Our results showed that 1203 incidents were reported in 641 of 1507 patients, of which 122 incidents were critical. 78. 3 % of all incidents were detect-ed by acute pain service stuff. The most common incidents included complica-tions, insufficient analgesia and problems with delivery circuits. Human factors were involved in 28. 9 % of the incidents, most being associated with technical failure due to unskillfu1ness, poor communications between APS stuff and pa-tients and lack of cooperation with surgeons and nurses. The general satisfactionrate of the patients was 90. 8 %. There was a very significant difference between the satisfaction of the patients who suffered from incidents and who did not (P

关键词： pain postoperative pain relief quality control incident

A Geometric Representation Scheme Suitable for Shape Optimization

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Computer Aided Drafting,Design and Manufacturing 1997年第2期7卷 66-72页

作者： Wang Xuelin and Zhou Ji CAD Center, huazhong university of science and technology, wuhan, Hubei 430074,P.R. china

A physical approach is used to provide shape modification of NURBS curve. The movement of the control polygon of NURBS is simulated by a physical frame structure. The shape of the design boundaries is described by a linear combination of n mode shape vectors, which are introduced by modal analysis. The control points of NURBS curve are then mapped into mode coordinate system, and the curve deformation is then carried out through the changes of mode coordinates. Numerical examples are used to demonstrate the applicability of this approach to structural shape optimization.

关键词： shape optimization NURBS curve finite element method

Piezoelectric and dielectric properties of Cr-doped PSN-PZN-PZT qua-ternary piezoelectric ceramics

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Rare Metals 2005年第2期24卷 185-189页

作者： SUNQingchi WANGLifeng LIUPing NIEQiang SchoolofMaterialsscienceandEngineering TianjinUniversityTianjin300072China SchoolofChemicalEngineeringandtechnology TianjinUniversityTianjin300072China

PSN-PZN-PZT + x wt. %Cr2O3, X = 0.0-0-9, were prepared by conventional mixed oxide techniques at sintering temperatures of 1220 degrees C-1300 degrees C for 2 h. The effect of sintering temperature on the microstructure and the piezoelectric properties was investigated by XRD, SEM, and other conventional measurement. The result indicated that with temperature increasing, the valence of Cr ion from Cr5+ or Cr6+ changes into C3+, and the piezoelectric properties turn hard. With increasing Cr2O3 content, the amount of rhombohedral phases increases and the morphotropic boundary phase is correspondingly shifts to rhombohedral phase. A uniform microstructure and excellent comprehensive properties were obtained at 1240 degrees C as the amount of Cr2O3 is 0.5 wt.%.

关键词： inorganic nonmetallic materials PSN-PZN-PZT conventional solid reaction process Cr2O3 content sintering temperature piezoelectric properties

FORMING PRINCIPLE OF TWO SIDE-DIRECTION BURR AND IT'S PREDICTION IN METAL CUTTING

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Chinese Journal of Mechanical Engineering 2001年第4期14卷 300-304页

作者： Wang Guicheng (School of Machinery Engineering,Jiangsu university of science and technology) 江苏理工大学江苏

The burr is one of the common phenomena occurring i n metal cutting operations The mathematical mechanical model of two side dir ection burr formation and transformation is established with plane stress strain theory,based on the orthogonal cutting The main laws of formation and change of the burr are revealed,and it is confirmed by experiment result,which first realizes prediction of the forming and changing of the two side direction burr in metal cutting operation.

关键词： Metal cutting Two side direction burr Mathe matical mechanical model Prediction Precision and ultraprecision machining

Scan chain design for shift power reduction in scan-based testing

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science china(Information sciences) 2011年第4期54卷 767-777页

作者： LI Jia 1,HU Yu 2 & LI XiaoWei 2 1 The School of Software,Tsinghua university,Beijing 100084,china 2 The Institute of Computing technology,Chinese Academy of sciences,Beijing 100190,china The School of Software Tsinghua University Beijing China The Institute of Computing technology Chinese Academy of Sciences Beijing China

Test power of VLSI systems has become a challenging issue *** scan shift power dominates the average test power and restricts clock frequency of the shift phase,leading to excessive thermal accumulation and long test *** paper proposes a scan chain design technique to solve the above *** on weighted transition metric (WTM),the proposed extended WTM (EWTM) that is utilized to guide the scan chain design algorithm can estimate the scan shift power in both the shift-in and shift-out ***,the wire length overhead of the proposed scan chain design can also be reduced by the proposed distance of EWTM (DEWTM) *** results confirm that the proposed approach can significantly reduce scan shift power with low wire length overhead.

关键词： low power DfT scan-based testing test power reduction scan chain design