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| 1 | A novel nonviral nanoparticle gene vector: Poly-L-lysine- silica nanoparticles显示文摘DNA delivery is a core technology for gene structure and function research as well as clinical settings. The ability to safely and efficiently targeted transfer foreign DNA into cells is a fundamental goal in biotechnology. With the development of nanobiotechnology, nanoparticle gene vectors brought about new hope to reach the goal. In our research, silica nanoparticles (SiNP) were synthesized first in a microemulsion system polyoxyethylene nonylphenyl ether (OP-10)/cyclohexane/ammonium hydroxide, at the same time the effects of SiNP size and its distribution were elucidated by orthogonal analysis; then poly-L-lysine (PLL) was linked on the surface of SiNP by nanoparticle surface energy and electrostatically binding; lastly a novel complex nanomate-rial-poly-L-lysine-silica nanoparticles (PLL-SiNP) wasprepared. The analysis of plasmid DNA binding and DNase I enzymatic degradation discovered that PLL-SiNP could bind DNA, and protect it against enzymatic degradation. Cell transfection showed that | Shiguo Zhu Hongbin Lu Juanjuan Xiang Ke Tang Bicheng Zhang Ming Zhou Chen Tan Guiyuan Li | 2002 | Chinese Science Bulletin2002，47，8： | 9 |
| 2 | Studies of poly-L-lysine-starch nanoparticle preparation and its application as gene carrier显示文摘Anion starch nanoparticle (StNP) with a diameter of 50 nm was prepared in wa- ter-in-oil microemulsion, with soluble starch as raw materials and POCl3 as crosslinking agent. PLL-StNP was prepared by linking poly-L-lysine (PLL) on the surface of StNP. At the same time, the size of PLL-StNP and its stability in aqueous solution were checked by AFM. The analysis of plasmid DNA binding, DNase I enzymatic degradation, toxicity and transfection were done. We discovered that PLL-StNP may be used as non-virus nanoparticle gene carrier. And we devel- oped the method of preparing PLL-StNP gene carrier and used it in cell transfection. As non-virus gene carrier, PLL-StNP has some advantages, such as large load of DNA, high transfection effi- ciency, low cell toxicity and biodegradability. | XIAO Suyao LIU Xuanming TONG Chunyi LIU Jun TANG Dongying ZHAO Lijian | 2005 | Science China Chemistry2005，48，2： | 7 |
| 3 | Deoxyribozymes inhibit the expression of periodl gene in vitro显示文摘To investigate the effect of two deoxyribozymes targeting period1 (per1) mRNA in vitro for exploring a novel gene therapy approach about circadian rhythm diseases, the specific deoxyribozymes targeting per1 were designed and synthesized chemically following MFold analysis according to its mRNA secondary structure. per1 RNA fragments were prepared by in vitro transcription of pcDNA3.1(+)-per1164:256. The cleavage reactions containing deoxyribozymes and per1 RNA fragments were performed under certain conditions. With the transfection tech- nique mediated by LipofectAMINETM, pcDNA3-per1 and DRz164 or DRz256 were introduced into NIH3T3 cells. The effects of deoxyribozymes on per1 were studied by reverse tran- script-polymerase chain reaction (RT-PCR) and flow cytometry (FCM). When deoxyribozymes and RNA transcripts were incubated under the adopted conditions at 37℃ for 2 h, about 63% of per1164:256 RNA transcripts were cleaved by DRz164 and about 50.5% by DRz256. After co- transfecting pcDNA3-per1 with DRz164 or DRz256, the expression of per1 mRNA was de- creased, as indicated by RT-PCR semi-quantity analysis. FCM analysis showed that Per1 protein was inhibited. Both DRz164 and DRz256 targeting per1 have the specific cleavage activity to- ward per1 mRNA in vitro and can highly block the expression of per1 gene in cellular milieu. | ZHOU Wei WANG Yueqi LIU Yanyou PENG Wenzhen XIAO Jing ZHU Bin WANG Zhengrong | 2005 | Science China(Life Sciences)2005，48，3： | 1 |
| 4 | The study of human PDGF-B gene transferred to cat corneal endothelial cells显示文摘AIM: To demonstrate that human platelet-derived growth factor-B (PDGF-B) cDNA could be expressed in primary cultured cat corneal endothelia cells by using gene transfer techniques; to explore a useful tool for the further studies of the molecular mechanisms of corneal endothelium failure and provide a potential effective genetic therapy for the blind patients. METHODS: Human PDGF-B cDNA was isolated from human placent by RT-PCR and inserted into pcDNA4 vector to construct recombinant eukaryotic expression plasmid pcDNA4-PDGF-B. The full length was confirmed by the DNA sequencing analysis. By tearing endothelium technique we obtained pure single layer of cat corneal endothelial cells. The pcDNA4-PDGF-B eukaryotic expression vector was transferred into cat corneal endothelial cells by EffecteneTM lipofectine. The transfection efficiency of EffecteneTM lipofectine in pcDNA4-B was detected with pcDNA4-GFP. 5 days later, RT-PCR was used to check the PDGF-B expression. Cell viability was tested by modified tertrozalium salt (MTT) method. Cell morphology was observed under inverted phase contrast microscope. RESULTS: The human PDGF-B cDNA was isolated successfully from healthy parturien placent tissue and the sequence was confirmed by computer automatic sequence and PCR analysis. Pure single layer cat corneal endothelial cells were successfully cultured by tearing endothelium technique. EffecteneTM lipofectine transfection technique could be effectively used to transfer pcDNA4-PDGF-B into cat corneal endothelial cells in vitro, the transfection efficiency was 30%. RT-PCR result showed that human PDGF-B gene was highly expressed in transfected cat corneal endothelial cells. The expressed PDGF-BB protein promoted the viability of cat corneal endothelial cells.CONCLUSION: Human platelet-derived growth factor-B (PDGF-B) cDNA could be highly expressed in cultured cat corneal endothelial cells by gene transfection techniques. Expressed PDGF-BB protein significantly promoted the viability of cat corneal endothelial cells, thus provided a potential effective method for corneal endothelium blindness genetic therapy. | Wen-Juan Luo, Chuan-Fu Wang | 2012 | International Journal of Ophthalmology(English edition)2012，5，1： | 1 |
| 5 | RETINOIC ACID NUCLEAR RECEPTORα (RARα),A MAJORROLE IN MEDIATING RETINOIDS INHIBITION OF GROWTH IN HU显示文摘RETINOICACIDNUCLEARRECEPTORα(RARα),AMAJORROLEINMEDIATINGRETINOIDSINHIBITIONOF GROWTHINHUMANBREASTCARCINOMACELLSShaoZhimin邵志敏;... | 邵志敏 沈镇宙 | 1995 | Chinese Journal of Cancer Research1995，7，2： | 0 |
| 6 | INHIBITED NEOPLASTIC PHENOTYPE BY THE c-Ha-ras ANTISENSE RNA显示文摘A 2.0 kb fragment DNA plasmid which expresses antisense to the upstream first exon of c-Ha-ras oncogene was transfected into Ha-ras transformed cell lines, GCM-3T3 and REF-4.3. The transfection leads to the inhibition of malignant behaviour, shown by decreasing of growth speed, colony forming ability on soft agar, tumorigenicity in nude mice and increasing ot differentiation degree. In GCM-3T3 cells the lung metastasis frequency became much less (from 60% to 12.5%) after the transfection and the expression of ras oncogene product, and p21 protein was obviously decreased in the transfected cells. This work has first shown the inhibitory effect of antisense RNA on neoplastic behaviour in China. | 路桂荣 武纯静 柯杨 熊辛 李延钧 刘景梅 许毅 胥薇 李吉友 邓国仁 鄂征 | 1991 | Science China Chemistry1991，34，12： | 0 |
| 7 | Functional expression of gap junction gene Cx43 and the myogenic differentiation of rhabdomyosarcoma cells显示文摘Rhabdomyosarcoma (RD) cells express low levels of the gap junction protein connexin 43 (Cx43), and its mRNA, and display very weak gap junctional intercellular communication (GJIC) as detected by Cx43 immunofluorescence, slot-blot and dye-transfer methods. These cells grow rapidly and show aberrant and incomplete myogenic differentiation. To investigate the role of gap junctions in these cells, the expression of Cx43 with relation to cell growth and myogenic differentiation in RD single-cell subclones-RDL3 and RDL6 is studied. The subclone RDL3 grows slowly and displays better myogenic differentiation. The expression of Cx43, its mRNA and the GJIC in RDL3 is comparable to that in normal myoblasts. Another subclone RDL6 which grows rapidly, but is poorly differentiated, expresses very low levels of Cx43 and its mRNA, and very weak GJIC. By using the calcium phosphate precipitate transfection technique, a full-length cDNA-encoding Cx43 and a pSV2neo have been introduced into the RDL6 cells. Several | 林仲翔 张志谦 韩亚玲 C.C.G.Naus K.R.Yu H.Holtzer | 1995 | Science China Chemistry1995，38，3： | 0 |
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