脂质体的

Methods Eukaryotic expression plasmid pCMVCD was constructed , and identified by re2 striction endoenzyme digestion. CD gene was transfected into NSCs from new2born Wistar rats using Lipofec2tamine2000. Positive clones (named NSCs/ CD cells) were screened by G418 presence. 52Fluorocytosine (52FC) ad2ministration of different concentrations were incubated with NSCs/ CD cells. NSCs/ CD cells viability ratios were mea2 sured by MTT assay.

通过构建真核表达质粒pCMVCD ，限制性内切酶消化鉴定后，采用Lipofectamine 2000 脂质体介导法转染新生大鼠室管膜下区神经干细胞(Neural stemcells ， NSCs)，G418 筛选阳性克隆，加入不同浓度的52氟胞嘧啶(52Flourocytosine ， 52FC)，MTT比色法测定NSCs的生存率。

Methods:Marrow stromal cells were obtained from the dual ilium of three adult dogs.

从狗的髂骨穿刺骨髓分离骨髓基质细胞，应用脂质体转染VEGF165基因，以RT-PCR检测转染的基因表达。

Interval observations under thefluorescent microscop were performed with excition waves of 470—490 nm.The results displayed that GFP genes were expressed effectively in E.octocarinatus, and GFP was distributed unifimly around macronuclearand diffused to the entire cell gradually. The chromosomes can be kept inthe macronucleus for 90h.

用优化的脂质体转化方法将含有人工染色体的重组质粒pBTub-tel2转化到处于有性生殖分裂间期阶段的游仆虫细胞中，结果表明，GFP基因在游仆虫细胞中得以高效表达，36-48小时绿色荧光均匀分布在细胞核周围，逐渐扩散到整个细胞，染色体能够在大核中保持约90小时。

Fluorescence in situ hybridization was carried out to confirm the integration of HBV DNA into male pronucleus and its replication with cell division in embryonic development.(Reverse transcriptase polymerase chain reaction,RT-PCR) and immunofluoresence assay were performed to observe the expression of the HBV gene in two-cell stage.

材料与方法：成熟雄鼠麻醉后双侧睾丸注射经脂质体DOSPER包裹的HBV质粒，手术后雄鼠与超排雌鼠合笼交配，用荧光原位杂变(Fluorescence in situ hybridization，FISH)分别检测单细胞胚和二细胞胚间期核中HBV DNA的存在与复制，用逆转录聚含酶链反应(Reverse transcriptase-polymerase chain reaction，RT-PCR)和免疫荧光方法检测HBV基因在二细胞胚胎中的表达。

METHODS: MyoD cDNA fragments were extracted from plasmids pEMSV-MyoD with polymerase chain reaction, and PCR was used to clone the whole-length gene of MyoD. After adding CACC sequence at 5' end, MyoD gene was cloned by orient topology into transfer ventor, pENTR/D-TOPO. Objective gene was transferred into adenoviral expression vector DNA via pENTR/D-TOPO vector. The recombinant adenoviral vectors transfected into HEK293A cells by using lipofectamine were packaged and amplified.

从pEMSV-MyoD质粒上用聚合酶链反应法扩增出MyoD cDNA片段，再通过聚合酶链反应使MyoD基因加上CACC序列接头，经过定向拓扑克隆使目的基因连接到转移载体上，再通过LR酶促反应，将目的基因转移到腺病毒表达载体DNA上，获得MyoD基因重组的腺病毒DNA，用脂质体转染法转染HEK293A细胞，包装扩增出MyoD基因重组的腺病毒。

These transgenic cells were able to secrete hEGF protein to certain extent, and have the biological activities to enhance the growth rate of Hacat cells in vitro. Conclusion: Adult epidermal keratinocytes in vitro transfected with exogenetic hEGF cDNA can express and secrete active hEGF.

质粒pcDNA3.1-hEGF在脂质体介导下成功转染成人皮肤角质形成细胞，转基因细胞能分泌有生物活性的EGF；体外培养的成人皮肤角质形成细胞可见少量EGF分泌。

The first -order rate constants for the transmembrane transport of the complxes into human erythrocytes were detemined and the mechanism was studied on the basis of the interactions of these complexes with cellular membranes and liposome by ESR,fluorescence technique, mercapto measurement and gelchromatography.

测定了它们跨人红细胞膜的一级反应动力学常数。用膜蛋白巯基滴定、荧光标记、自旋标记ESR谱等方法研究了它们与膜蛋白、膜脂的相互作用；用脂质体模拟了这些配合物的细胞摄入。

The therapy groups displayed significant improvements in neuromotor functions and survivals of neuron and oligodendrocyte compared with control groups. These results demonstrated the neuroprotect roles of GGF2, and cationic liposome-mediated GGF2 transgene therapy has potential possibility.

结果：1、大鼠LFP致伤后3—7天，GGF2 mRNA在海马CAl区、CA2区、CA3区、DG及皮层表达明显增加。2、阳离子脂质体介导GGF2转基因治疗可促进大鼠伤后行为学的恢复，可促进损伤区神经元及少突胶质细胞的存活。3、原核表达方法得到GGF2重组蛋白。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

Results: it was efficient to transfect chicken embryonic fibroblast cells by the method of lipofectamine mediation. the expression levels of protein were significantly different.

结果 脂质体介导的方法能有效转染鸡胚胎成纤维细胞，实验中egfp蛋白相对表达量随质粒量的增加而明显升高（p.01）。

The basic concept of FOP can be summarized as to further optimize effective prescription according to the standard of curative effects and with the aid of modern science and technology and theories of traditional Chinese medicine.

其基本内涵可概括为：以确有疗效的中药复方为研究对象，以现代科学技术和传统中医药理论为技术支持，以该复方所治病证的药效响应为评价标准，以优化重组疗效更优的新复方为研究目的。

Ever since our world has been a world, native forests have been indiscriminately exploited by man.

自从我们的世界一直是世界原生森林被任意剥削人。