脂质体

Wtp53 has obvious antiblastic effect towards Y79 cells, and the apoptosis rate of Y79 cells was the highest after transfection mediated by ultrasound-microbubble.

结论wtp53基因对RB瘤细胞具有较明显的抑制生长的作用；质粒+微泡+超声辐照，质粒+脂质体转染，以及质粒+超声辐照，均可促进Y79细胞的凋亡，但超声微泡介导的转染引起的凋亡率高于脂质体转染，而二者的凋亡率又明显高于质粒+超声转染。

Homogeneous pressure and the number of passes through the homogenizer affect the entrapment efficiency and average particle size in different degrees.

以乳脂肪球膜磷脂为膜材，采用高压均质法制备免疫球蛋白G脂质体，研究了不同磷脂浓度、不同均质压力和不同均质次数对脂质体的包埋率和平均粒径的影响。

The formulation was optimized on the basis of orthogonal design and its entrapment efficiency was performed by the protamine sedimentation method .

方法采用薄膜蒸发一高压均质法制备槲皮素脂质体，通过正交设计优化处方工艺，以鱼精蛋白沉淀法分离脂质体与游离药物，测定药物的量。

HRP was coupled with the fluorochrome FITC and encapsulated with liposome . pEGFP-N1 plasmid (expressing green fluorescent protein) with encapsulation in commercially available liposome was prepared. The eyes were enucleated 1, 4 and 8 weeks after zonule rupture and anterior segments comprising lenses were incubated in medium containing one of these components. Cryo-sections were made and translocation of fluorescent macromolecule from the medium into the lens and green fluorescent protein expression in the epithelium were observed by fluorescent microscopy.

制备FITC标记的HRP脂质体；制备pEGFP-N1质粒（表达绿色荧光蛋白的质粒），并用脂质体包裹；豚鼠悬韧带部分离断后1周、4周、8周取含晶状体的眼前段标本分别在含有FITC-HRP复合物、pEGFP-N1质粒的培养基中孵育，冰冻切片，荧光显微镜观察FITC-HRP复合物进入晶状体和pEGFP-N1质粒在晶状体内表达的情况，比较悬韧带离断侧与非离断侧的差异。3。

Chapter two, the preparation of liposomes in DPPC/Chol/DPPA mixed system were investigated systematically for the first time. The optimal conditions for preparation and the composition of liposomes were established, the encapsulation efficiency was up to 50.4%.

第二章 首次系统研究了反相蒸发法用于DPPC/Chol/DPPA混合类脂体系的脂质体的制备，确定了脂质体的最佳制备条件和最佳组成，获得了高达50.4％的包裹效率。

After the cell growth curves was recorded, RPE cells of the 3-5th passages were utilized. 2、Three different siRNA (siRNAl,siRNA2,siRNA3) targeting against human cx43 gene and one negative control siRNA were designed and transfected into cultured human RPE cells via liposome reagent. The most effective siRNA can be determined by semi-quantitative reverse transcription PCRRT-PCR. 3、To the most effective siRNA, after transfected into human RPEs with different concentration, the cellular proliferate activities were messured by MTT colorimetry ; the percentages of RPE in different cell circle phase was assayed by FCM; the changes of phenotypical properities were observed with SCM; the protein expression of cx43 was studied through immunocytochemistry stain and Weston blot; the communication intercellular was calculated with FRAP; and the ability of recovery was assessed by using an in vitro wound healing model.4、The total proteins of siRNA1 and RPE were seperated by two-dimensional gel electrophoresis and visualized by silver staining. Proteins with significant expression alterations were selected and their peptide mass fingerprints (PMFs were obtained by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).The PMFs were used to search NCBInr database by Auto MS-Fit software.

实验方法：1、培养原代的人RPE细胞，经过细胞角蛋白、S-100和神经胶质原纤维酸性蛋白免疫细胞化学鉴定后，通过AO/PI染色技术确定培养细胞的存活率，描记其生长曲线，第3-5代用于以下细胞实验2、生物合成针对人cx43基因的三条小干扰RNA和一条阴性RNA通过脂质体转染RPE细胞后，通过RT-PCR的方法确定抑制效率最高的干扰片断3、将该片段以不同浓度通过阳离子脂质体转染培养的人RPE细胞后，采用MTT法观察其对细胞的增殖力的作用；通过流式细胞仪观察其对细胞周期的影响；通过扫描电镜观察其对细胞形态的影响；通过免疫细胞化学和Weston blot观察其对cx43蛋白表达的作用；采用激光共聚焦和荧光淬灭恢复技术观察荧光恢复速率平均百分率，评价其对细胞间通讯功能的影响；通过制作RPE细胞损伤模型，观察其对损伤修复能力的作用4、分离纯化转染siRNA的RPE组和正常对照组RPE细胞的全部蛋白质，应用等电聚焦电泳和SDS-PAGE双向电泳技术，银染显示分离出的蛋白质斑点，经凝胶图像分析软件对两个样本进行胶图分析，寻找差异蛋白点。

After light radiation time of one hour,the IC50 of cytotoxicity of C60 liposomes is 154.9 μg/ml.During the concentration of C60 liposomes are 16.86 μg/ml and 33.72 μg/ml separately,the plaque reduction rate of direct virucidal activity to HSV-1F strain is positive correlation to the time of light radiation,the correlation coefficients rate 0.8510（P.005） and 0.9766 （P.005） respectively.

光照1 h，C60脂质体对Vero细胞的细胞毒为154.9 μg/ml.C60脂质体浓度为16.86 μg/ml及33.72 μg/ml时，对HSV-1F株直接杀灭的空斑减数率与光照时间成正相关，相关系数r分别为0.8510（P.005）和0.9766（P.005）；当C60脂质体浓度为16.86 μg/ml时，对HSV-1F株感染阻断的空斑减数率与光照时间正相关，相关系数r依次为0.84（P.005）及0.9923（P.005），说明C60的抗病毒作用具有时间效应，主要是光敏动力学机制起作用。

The CD59 gene and MCP gene were mixed with liposome, then used to produce transgenic mice by 3 methods. Firstly, the gene/ liposome complex was directly injected into 5 male mice's testis. The 5 transfected males copulated with 25 females each 7 days later.

将分别含CD59基因和MCP基因的表达载体与脂质体混合制成基因-脂质体复合物，分别采用睾丸注射、输精管注射和精子/基因-脂质体复合物共孵育三种方法生产转双基因小鼠。

Results Brucine and its liposome had significant inhibiting effect on transplanted tumor growth in mice. Inhibitory rates of brucine on Heps were 35.05% [1.61mg/], 43.70% [3.23mg/], and 46.09% [6.46mg/], while of its liposome on Reps was 45.41% 1.61mg/1 and 58.19% [3.23 mg/, respectively. The inhibitory rate of brucine on S180 was 37.59% [1.61 mg/], 36.13% 3.23mg/, and 38.87% [6.46 mg/, while of its liposome on S180 was 53.00% [1.61mg/] and 48.37% [3.23mg/], respectively. However, both of them had no distinct effect on prolonging the life of mice with transplanted tumor.

结果 马钱子碱及其脂质体能有效抑制实体瘤模型荷瘤小鼠体内肿瘤生长，1.61、3.23、6.46mg/马钱子碱对Heps小鼠的抑瘤率分别为35.05%、43.70%和46.09%，而1.61、3.23mg/马钱子碱脂质体的抑瘤率分别为45.41%和58.19%；1.61%、3.23、6.46mg/马钱子碱对S180小鼠的抑瘤率分别为37.59%、36.13%和35.57%，而1.61和3.23mg/马钱子碱脂质体的抑瘤率分别为53.00%和48.37%，明显高于马钱子碱；但两者对腹水瘤模型荷瘤小鼠的生存时间均无延长作用。

METHODS: Lipofectamine method: The BMMSCs were obtained from the tibias and femurs of the guinea pigs. The third passage BMMSCs were cultured with plasmid-lipofectamine mixture for 6 hours, followed by fetal bovine medium for 48 hours. Immunohistochemistry was performed for transient expression. G418 was added after 48 hours.

①脂质体法：取体外分离、培养的第3代豚鼠骨髓间充质干细胞，将质粒-脂质体混合物加入含细胞的培养基中培养6 h，再加入胎牛血清的培养基，孵育48 h 后行免疫组织化学检测，即为瞬时表达。48 h后加入含G418培养基筛选。

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。