脂质体

Methods: Genes of MCP and DAF were connected by 10aa coding sequence of enriching Gly and Ser. Then insert into pcDAN3.1 vector, constituting pcDNA-MD3 expression plasmid of hMCP-DAF fusion gene. The genes were transformed by liposome into the pig endothelial cell; human serum handles masculine cloning, identify the function that restrain breaking of person alexin.

将MCP、DAF两个基因以富含Gly和Ser的10aa编码序列作铰链串接，并接入表达载体pcDNA 3.1中，构成含hMCP-DAF的真核表达质粒(pcDNA-MD3)；以脂质体转染猪内皮细胞，人血清处理阳性克隆细胞，鉴定抑制人补体溶破的功能；应用显微注射将线性基因导入猪受精卵；PCR检测导入基因整合率。

GFP gene eukaryotic express plasmids were transfected into ES cells. The cells were observed under the inverted fluorescent microscope. The clones which expressed the powerful green fluorescence were chosen to be named ES-GFP cells and cultured continuously. The examination of ES cell totipotent including Alkine phosphatase staining, embryonic body formation in vitro and teratomas formation in nude mice was implemented.

1。采用脂质体方法将GFP质粒转染ES-D3细胞株，经筛选后倒置荧光显微镜观察细胞形态及荧光状态，挑取荧光最强的克隆，命名为ES-GFP细胞，进一步扩增培养，观察细胞生长及荧光表达情况，行细胞分化全能性鉴定包括碱性磷酸酶（alkine Phosphtase，AKP）染色、体外胚胎体（embryonic body，EB）形成实验及裸鼠体内成瘤实验。2。

Anisodamine causes interdigitation in DPPG, whose phospholipid molecules insert into the opposite lipid layer with the ends of the acyl chains near to the 5th carbons of the opposing lipid molecules, whereas anisodamine can not cause interdigitation in DPPC existed generally in membranes.

山莨菪碱诱导DPPG脂质体交插结构，其脂酰链末端插到对面分子层脂酰链第五个碳原子的位置，而生物膜中普遍存在的DPPC不能被山莨菪碱诱导形成交插相，但DPPG／DPPC混合物则能形成交插相，即伴随DPPG的交插，DPPC分子也发生交插。

The expression of CD59 on the cell membrane was tested by cell immunohistochemistry and flow cytometry. Results pALTER plasmid containing CD59 was cut with restriction enzymes and a 496 bp fragment obtained by electrophoresis, which was complete conformity with the insert.

将两种含有不同突变体的人CD59全长cDNA序列重组pALTER质粒，应用阳离子脂质体导入法与pcDNA共转染CHO细胞，用G418筛选阳性克隆，应用细胞免疫组化及流式细胞仪检测CD59在CHO细胞表面的表达。

Methods: Two recombinant palter plasmid containing whole CDNA sequences of HMCD59 transfected CHO cells by lipofectamine with PCDNA respectively. Then G418 was used to select positive clone.

分别构建两种含有HMCD59全长CDNA序列的重组pALTER质粒，并与pCDNA3质粒，按3：1比例混合后，运用脂质体介导法共转染CHO细胞(编号为HM5-CHO及HM6-CHO)，用新霉素类似物G418初步筛选出阳性克隆。

Inaddition,we extract total RNA from cultural U251 cell of humanglioma,make coding gene extron amplification of TfR through ReverseTranscription-Polymerase Chain Reaction,construct expressionplasmid of pCDNA3.1-TfR after digestion of enzyme andconjunction,do transfection to U251 cell by liposome afteridentification,cause excess expression,detect the efficiency of transientexpression of TfR protein by pEGFPN1-TfR, sieve positive cell ofpCDNA3.1-TfR through G418, choose positive cell clone, detect theexpression of TfR in anteroposterior period through Western blot,FCM,cell immunochemistry and mRNA through RT-PCR, identify the effect of transfection.

另外，培养人脑胶质瘤U251细胞，进行细胞总RNA抽提，运用RT-PCR方法进行TfR基因的完整编码区外显子的扩增，经过酶切、连接后构建pCDNA3.1-TfR的表达质粒，鉴定正确后按脂质体转染方法转染人胶质瘤U251细胞，使其过量表达，通过pEGFPN1-TfR检测其瞬时转染的效率；用G418对稳定表达pCDNA3.1-TfR的细胞进行筛选，挑选阳性细胞克隆，运用Western blot、流式细胞检测技术和细胞免疫化学以及mRNA半定量的方法检测TfR在转染前后的表达水平，鉴定转染的效果。

The pcDNA〓-apoE〓 plasmid was transfected to SK-N-SH Neuroblastoma cell by liposome. It could be screened by G〓. The immunohistochemical assay was shown that neuroblastoma cell transfected by pcDNA〓-apoE〓 expressed the recombinant apoE protein. The intracellular expressed recombinant protein could inhibit the death of neuroblastoma cell induced by beta amyloidal peptides 25-35 fragment.

用pcDNA〓—apoE〓真核表达载体与脂质体共转染神经成纤维胶质瘤细胞，用G〓选择压力筛选，细胞免疫荧光检测表明，pcDNA〓—apoE〓转染的神经成纤维胶质瘤细胞表达了apoE〓重组蛋白，这种内源性的apoE〓重组蛋白对25μM Aβ25—35多肽诱导的神经成纤维胶质瘤细胞的死亡具有拮抗作用。

Encapsulated psoralen were evaluated by dialyzation using psoralen gel as the control. Changes of liposome????encapsulated psoralen and release rate were detected during a 3????week storage at 4 ℃ for evaluating the stability of the liposomal formulation.

以同浓度的补骨脂素凝胶为对照，用透析法检测补骨脂素脂质体凝胶的体外释药模式，并对其在4 ℃下贮存3周的释药稳定性进行研究。

All the transfected and un-transfected rMSCs were attached to Allo-DBMs. These new biomaterials were implanted in muscle bags and segmental radius defects of the New Zealand white rabbits, and some controlled material groups were established for comparison. All the biomaterials and the controlled materials were assessed by gross observation, radiographical and histological methods.

取兔自体骨髓基质细胞培养扩增，用脂质体介导的方法转染人骨发生蛋白2基因，将转染和未转染人骨发生蛋白2基因的自体骨髓基质细胞附和于异体兔脱钙骨基质形成新的生物植骨材料，连同空白及单纯脱钙骨基质对照组植入兔前肢肌袋和桡骨缺损。2、4、6、8周分别取材进行大体，放射线和病理观察。

Changes of liposome encapsulated psoralen and release rate were detected during a 3 week storage at 4 ℃ for evaluating the stability of the liposomal formulation.

以同浓度的补骨脂素凝胶为对照，用透析法检测补骨脂素脂质体凝胶的体外释药模式，并对其在4 ℃下贮存3周的释药稳定性进行研究。

The labia have now been sutured together almost completely.The drains and the Foley catheter come out at the top.

此刻阴唇已经几乎完全的缝在一起了，排除多余淤血体液的管子和Foley导管从顶端冒出来。

To get the business done, I suggest we split the difference in price.

为了做成这笔生意，我建议我们在价格上大家各让一半。