Production of human lysozyme-transgenic cloned porcine embryos by somatic nuclear transfer

Due to their physiology and organ size, pigs have significant potential as human disease models and as organ transplantation donors. Genetic modification of pigs could provide benefits for both agriculture and human medicine. In this study, five fetal pig fibroblast cell lines from two species (Wuzhishan and Landrace pigs) were transfected using double-marked human lysozyme (HLY) plasmids (pBC1-HLY-GFP-NEO) by a liposome-mediated method. The ratio of green fluorescent protein (GFP)-expressing cells was >95% in sw7, sw8, slw3 and slw6 cell lines, but only 49.3% in slw9 cells. Cells from the four highly transgenic lines were used as nuclear donors to construct embryos, which were then cultured after fusion and activation by electric stimulation. The rate of cleavage was 76.7%, 48 h after activation. After 7 days, 18.5% of cleaved eggs had developed to the blastocyst stage and 93.3% of blastocysts were GFP-positive. These results indicate that transgenic fetal pig fibroblast cell lines could be obtained by a liposome-mediated method, though the transfection efficiency varied between cell lines. Reconstructed embryos derived from transgenic cells could successfully develop into blastocysts, most of which were GFP-positive.     

吐温-80和脂质体对厚朴酚荧光光谱的影响

分析厚朴酚在吐温-80胶束和脂质体溶液中荧光光谱的变化,发现：1）浓度为2mmol·L^-1的吐温-80胶束对厚朴酚有荧光增敏作用,厚朴酚存在于吐温-80的疏水核中,其荧光光谱发生了明显的红移;2）在4mg·mL^-1的脂质体溶液中,厚朴酚的荧光光谱变得复杂,谱图信息反映厚朴酚同时存在于水相和双脂层膜相中,且在二者之间存在分配平衡,通过测定λ365、λ397处的荧光光谱随厚朴酚浓度的变化,得到了厚朴酚在脂质体膜相和水相中的分配平衡关系;3）脂质体溶液中加入吐温-80,厚朴酚在双脂层膜相中的分布增加,其荧光光谱发生了红移．     

细胞内磁热疗研究概述

热疗即加热疗法,是指通过升高肿瘤温度,利用热能杀灭癌细胞和肿瘤组织的一种物理疗法．通过在磁性纳米粒子表面进行各种修饰,提高磁性纳米粒子进入肿瘤细胞的效率及选择性,在体外交变磁场的作用下促使进入细胞内的纳米磁性粒子产热而杀伤肿瘤细胞,这种方法称为细胞内磁热疗．作者就肿瘤细胞内磁热疗的研究进展作一综述．     

脂质体转移因子的制备及其活性研究

以转移因子为包裹对象,探讨提高脂质体肽类药物的包封率和减缓渗漏率的方法和条件,以及制备过程对活性的影响。采用逆相蒸发法制备、以正交试验设计筛选最佳配方组合,制备负电荷脂质体转移因子。通过巨噬细胞吞噬试验和E-玫瑰花结试验比较不同剂型、不同给药途径对小鼠免疫功能的影响。结果表明：负电荷脂质体转移因子包封率和稳定性均比其他文献报道的有较大的提高,包封率5次平均达43%,30d渗漏率12%,具有一定的稳定性;E-玫瑰花结试验显示：制备过程对转移因子活性无显著影响;巨噬细胞吞噬试验表明：不论注射或口服脂质体转移因子,均有提高机体免疫功能的作用,脂质体转移因子的药效优于游离转移因子。     

脂质体转染人肝星状细胞和肝细胞条件优化

利用Lipofectamine 2000(Lipo)介导LacZ及EGFP基因转染人肝星状细胞系LX-2和人肝细胞系Chang Liver,探讨Lipo用量、质粒用量、转染时间、细胞初始接种密度及不同启动子驱动对转染效率的影响.结果显示,两种细胞均能得到高效转染,Chang Liver细胞转染效率(80%)高于LX-2细胞(60%).Lipo用量为每孔2μL,转染后6 h换液,LX-2细胞应用质粒DNA为每孔0.45μg,Chang Liver细胞应用质粒DNA为每孔0.30~0.45μg,此时转染效率最高.巨细胞病毒(CMV)启动子驱动LacZ转染效率与CMV早期增强子/鸡β肌动蛋白(CAG)启动子驱动增强型绿色荧光蛋白(EGFP)转染效率无显著差异;细胞初始接种密度对转染效率无显著影响.     

脂质体磷脂与抗体蛋白的偶联方法

为使蛋白质分子(如抗体)易于在脂质体表面发生交联,同时能很容易地控制表面覆盖的范围,文中探讨一种简单而易于控制的免疫脂质体制备方法．通过添加一定比例的直链醛,并与卵磷脂、胆固醇混合,采用逆相蒸发法制备脂质体．研究结果表明,双层脂质分子中直链醛(十一醛)的亲水醛基暴露在水相中,能与抗体蛋白中的氨基形成席夫碱,并能通过改变最初与脂质混合时所使用的醛量来改变表面覆盖的范围．所制备的免疫脂质体抗体活性大大提高．     

Cypate介导的靶向热敏纳米脂质体用于肿瘤成像和光热触发的药物释放

本文合成了一种新型多功能的纳米靶向脂质体药物递送系统,即所述的光热敏感脂质体（PTSL）,它的水溶性层掺杂了阿霉素作为治疗剂,同时它的疏水层包裹了Cypate作为诊断剂外层磷脂修饰了靶向穿膜肽H_4-1.测试了PTSL的物理性质,稳定性以及脂质体的装载性能,合成的纳米复合材料具有针对性,稳定性和精准性,具有载药量（DL）9±1.5%和包封率（EE）82.7±2.1%.其Tm值落在43.1℃,其温度可以被认为能够显示有效的热疗法,但不燃烧的正常组织的合适温度.提高脂质体的Tm温度达到热敏脂质体的热敏温度,脂质体崩解同时包裹的化疗药物释放.与此同时,在细胞器水平上,PTSLs主要位于细胞质.当暴露在近红外光下,所述的Cypate可以产生单线态氧,从而引起膜的破裂.该脂质体可以实现靶向输送,增强光化学内化释放,化疗和热疗的功能.实验表明该PTSL是一种有前途的药物递送系统,用于肿瘤的诊断和靶向治疗.     

A new method for uniform local heating deep in body using ultrasound phased-array system

A new method for targeted heating of deep tissue was developed by using an ultrasound phased-array system which can generate various multiple foci patterns by electronically changing its amplitude or phase pattern. This method involves using a technique of combining switching and rotating of multiple foci patterns to create a uniform temperature over tissue volumes in various size. Using this method, the target tissue deep in the body can be heated to a specified temperature, which gives conditions for thermo-sensitive liposomes release. A simulation study for a 108-element, spherically sectioned array was performed to determine an optimal heating scheme from a set of multiple focus fields which were produced by inputting different combinations of phases and amplitudes. Comparisons of a static multiple foci field, the switched fields and the switched-rotated fields indicated that the technique of combining switching and rotating of multiple foci patterns has advantages of both lowering the peak temperature and evening the temperature distribution. The simulation results also show that the therapeutic heating zones in various size employing the combined method. These results offer significant data for designing thermotherapy equipment for tumor-specific drug release with thermo-sensitive liposomes.     

来源不同的卵磷脂脂质体载药性能的研究

本文研究了三种不同来源的卵磷脂(蛋黄卵磷脂,大豆卵磷脂、猪脑卵磷脂)脂质体的重要载药性质:包封率、渗漏。进一步测定了三种脂质体的粘度分布,微粘度及氧化指数,並研究了脂质体结构参数与其载药性的关系。实验证明,三种卵磷脂中,蛋黄卵磷脂制成的脂质体药药性能最佳。     

可聚合脂质体的研究 Ⅰ．含共轭双炔基团的两杂性单体及其脂质体

可聚合脂质体的研究Ⅰ．含共轭双炔基团的两杂性单体及其脂质体李子臣，何巍，李福绵（北京大学化学系北京１００８７１）关键词１０，１２－二十二碳二炔－１，２２－二酸（ＤＤＣＡ），脂质体，１０，１２－二十二碳二炔－１，２２－二醇（ＤＤＯＬ），光聚合聚合型脂质...