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| 1 | Inhibition of human La protein by RNA interference downregulates hepatitis B virus mRNA in 2.2.15 cells显示文摘AIM: To investigate the role of human La protein in HBV mRNA expression.METHODS: Three human La protein (hLa) specific siRNA expression cassettes (SECs) containing U6+1 promoter were prepared via one-step overlapping extension PCR. After transfection with SECs into HepG2 cells, inhibition effects on hLa expression were analyzed by semi-quantitative RT-PCR and Western blotting. Then, effective SECs were screened out and transfected into 2.2.15 cells, a stable HBV-producing cell line. HBV surface antigen(HBsAg) and e antigen (HBeAg) secretions into culture media were detected by microparticle enzyme immunoassay (MEIA) and HBs and HBe mRNA levels were analyzed by semi-quantitative RT-PCR.RESULTS: SEC products containing U6+1 snRNA promoter,and 3 sites of hLa mRNA specific siRNA were obtained successfully by one-step overlapping extension PCR and could be directly transfected into HepG2 cells, resulting in inhibition of La protein expression in both mRNA and protein levels, among which U6+l-hLa833 was the most efficient,which reduced 18.6-fold mRNA and 89% protein level respectively. In 2.2.15 cells, U6+l-hLa833 was also efficient on inhibition of hLa expression. Furthermore, semi-quantitative RT-PCR showed that HI3s and HBe mRNA levels were significantly decreased by 8-and 66-fold in U6+l-hLa833 transfected cells compared to control. Accordingly, HBsAg and HBeAg secretions were decreased partly posttransfection with SECs.CONCLUSION: PCR-based SECs can be used to mediate RNAi in mammalian cells and provide a novel approach to study the function of La protein. The inhibition of La protein expression can result in a significant decrease ofHBV mRNA, which implies that the hLa protein is also involved HBV RNA metabolism as one of the HBV RNA-stabilizing factors in human cells. | QinNi ZhiChen Hang-PingYao Zheng-GangYang Ke-ZhouLiu Ling-LingWu | 2004 | World Journal of Gastroenterology2004，10，14： | 13 |
| 2 | TESTING DIFFERENT CONJUGATE GRADIENT METHODS FOR LARGE-SCALE UNCONSTRAINED OPTIMIZATION显示文摘In this PaPer we test different conjugate gradient (CG) methods for solving large-scale unconstrained optimization problems.The methods are divided in two groups:the first group includes five basic CG methods and the second five hybrid CG methods.A collection of medium-scale and large-scale test problems are drawn from a standard code of test problems.CUTE.The conjugate gradient methods are ranked according to the numerical results.Some remarks are given. | Yu-hongDai QinNi | 2003 | Journal of Computational Mathematics2003，21，3： | 10 |
| 3 | Inhibition of hepatitis B virus surface antigen expression by small hairpin RNA in vitro显示文摘AIM: To explore the anti-hepatitis B virus effect of RNA interference (RNAi) using small hairpin RNA (shRNA)expression vector.METHODS: Hepatitis B virus surface antigen green fluorescent protein (HBs-GFP) fusion vector and shRNA expression vectors were constructed and cotransfected transiently into HepG2 cells. mRNAs extracted from HepG2 cells were detected by real-time PCR. Fluorescence of HBs-GFP protein was detected by fluorescence-activated cell sorting (FACS). The effective shRNA expression vector was transfected into HepG2.2.15 cells. HBsAg and HBeAg in HepG2.2.15 cells were analyzed by radioimmunoassay (RIA) method.RESULTS: FACS revealed that shRNA targeting at HBsAg reduced the GFP signal by 56% compared to the control.Real-time PCR showed that HBs-GFP mRNA extracted from HepG2 cells cotransfected with pAVU6+27 and HBs-GFP expression plasmids decreased by 90% compared to the empty vector control. The expressions of HBsAg and HBeAg were also inhibited by 43% and 64%, respectively.CONCLUSION: RNAi using shRNA expression vector can inhibit the expression of HBsAg, providing a fresh approach to screening the efficient small interfering RNAs (siRNAs). | Zheng-GangYang ZhiChen QinNi NingXu Jun-BinShao Hang-PingYao | 2005 | World Journal of Gastroenterology2005，11，4： | 8 |
| 4 | 显示文摘 | Fei Qinni Liu Yanhua Gu Tieeheng | 2011 | Journal of Lu- minescence2011，131，： | 1 |
| 5 | 藏木和八子的田园下午茶显示文摘"悠闲的午后,阳光正好时,可以自己一个人慵懒地躺着,可以与好友相伴谈天说地……总之,做自己喜爱的事最理想不过了。
我想要这样一个理想的下午,坐在自己的"小田园"里,与呆萌可爱的兔子和美丽飘香的花草作伴。喝一口茶,做着一款款漂亮精致的手工。" | 藏木Qinni | 2018 | 课堂内外(初中版)2018，，4： | 0 |
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