银杏内酯B柔性纳米脂质体的制备及体外透皮研究

目的：制备银杏内酯B(GB)柔性纳米脂质体，并对其体外透皮规律进行研究。方法：采用薄膜分散法制备银杏内酯B柔性纳米脂质体，并对其形态及粒径大小进行分析；采用改良的 Franz 扩散池进行体外透皮吸收试验，比较银杏内酯B醇溶液、银杏内酯B柔性纳米脂质体及普通银杏内酯B纳米脂质体的经皮累积渗透量及渗透速率。结果：此方法制得的脂质体平均包封率为(89.52±1.76)%，平均粒径为(208.3±25.49) nm，Zata电位为-49.2 mV。柔性纳米脂质体8 h的累积透过量为189.97 μg·cm^-2，8 h的渗透速率为23.75 μg·cm^-2·h^-1。结论：柔性纳米脂质体包封率较高，稳定性良好，可显著促进银杏内酯B的透皮吸收。     

脂质体包裹反义核酸对调控效果和降解时间的影响

目的 观察脂质体Dotap包裹反义脱氧寡核苷酸对酪氨酸酶(TYR)的调控效果和降解时间的影响,寻找适合于临床应用的高效转染、低降解率的载体和应用方法。方法 应用DNA合成仪合成针对TYR的反义脱氧寡核苷酸,32P标记、脂质体包埋,加入黑色素细胞培养体系进行点杂交,行黑素含量、酪氨酸酶mRNA表达量检测以及降解时间的同位素液闪检测。结果 经脂质体包埋的反义脱氧寡核苷酸使细胞黑素含量由(64±2.3)pg/cell下降至(23±1.5)pg/cell,并可使基因表达水平由160%降低至82%;同位素液闪显示经脂质体包埋的反义脱氧寡核苷酸能使其半衰期由8 h延长至12 h。结论 脂质体可以明显增强反义核酸的调控效果,并可延长其作用半衰期。     

The Experiment of Gene Therapy for Gallbladder Carcinoma Cell in Vitro with Herpes Simplex Virus Thymidine Kinase/Ganciclovir

Objective To evaluate preliminarily the effect of HSV - tk/GCV system on gallhladder carcinoma cells in vitro.Methods Recombinant retroviral vector PLtkSN containing tk suicide gene was transfacted into gallbladder carcinoma cells, mediated by LipofectAMINETM 2000 liposome, and the growth - inhibiting rotes of GBC, SD/tk cells in the different GCV concentrations were measured by MTr methods; GBC - SD cells were mixed in the different proportions, and death rotes of the mixed cells treated with GCV were detected to verify by stander effect. Resuts GCV could lead GBC - SD/tk cells to significant death, and there was striking difference compared with the control cells （p〈0.01）. When the concentration of GCV was 1,10, 50,100,500ug/ml, the killing rate of GBC - SD/tk cells was respecfively 6.8%, 2.5.2%, 54.5%, 66.3%, 89.3%, indicating dose - dependent phenomenon; With GBC - SD/tk cells in the proportion of 0, 10%, 20% ,50%, 70%, 100%, the killing rate of the mixed cells treated with GCV was 0, 19.7%, 40.3%, 77.7%, 88.0%, 93.5%, respectively. It was apparent that bystander effect was observed in the experiment. Condusion HSV - tk/GCV system could be a potential tool for the treatment of carcinoma.     

Cholera Toxin B-Mediated Targeting of Lipid Vesicles Containing Ganglioside GM1 to Mucosal Epithelial Cells

Purpose. To determine whether the non-toxic pentameric B subunit of Cholera toxin (CTB) binding to ganglioside GM1 on both the lipid vesicles and epithelial cells may provide a means to target lipid vesicles to mucosal cells expressing surface GM1. Methods. Sonicated lipid vesicles containing ganglioside GM1 were prepared. Inter-vesicle cross-linking due to pentameric CTB binding to these GM1 vesicles was determined with a sub-micron particle analyzer. Association of CTB to GM1 vesicles was analyzed with continuous sucrose gradient centrifugation. CTB-mediated binding of GM1 vesicles to human mucosal epithelial cells (Caco-2 and HT-29), mucous membranes of mouse trachea, and nasal tissues were detected with fluorescent labeled vesicles. Results. An increase in lipid particle size due to binding of CTB to lipid vesicles and inter-vesicles cross-linking was detected. At a 30-to-1 mole ratio of membrane-bound GMl-to-CTB, optimum increase in GM1 vesicle aggregation, was detected. Under such conditions, all the added CTB molecules were associated with GM1 vesicles. Time course analysis showed that inter-vesicles cross linking by CTB was detectable within 10 min. and reached a maximum value at 60 min. CTB associated GM1-vesicles bind to mucosal epithelial cells HT-29 and Caco-2 with similar affinity [K_d = 7.8 × 10^–4 M lipid (Caco-2) and 7.6 × 10^–4 M lipid (HT-29)]. GM1 mediated binding specificity was demonstrated by blocking with anti-GMl antibody and the insignificant degree of CTB-associated GM1 vesicle binding to GM1 deficient C6 cells. Conclusions. The CTB-mediated GM1 binding to multiple membrane surfaces provides selective localization of GM1 vesicles to GM1 expressing mucosal cells and tissues. The strategy may be useful in localizing drugs and proteins to gut and respiratory tract mucosa.     

弹性义齿修复的临床效果观察

目的：了解弹性义齿修复的临床效果，为临床应用提供参考。方法：152例门诊缺牙患者，制作弹性义齿修复体188件，上颌130件，下颌58件。Kennedy一类缺损修复6件，Kennedy二类缺损修复4件，Kennedy三类缺损修复102件，Kennedy四类缺损修复76件。通过1－4年的观察，从形态和功能、固定和稳定、颜色和美观以及对组织保健判断对义齿的满意程度，并进行临床效果评价。结果：79例95件弹性义齿具有良好的美观效果，颜色满意，感觉舒适，满意和较满意率达100％，Kennedy三、四类缺损修复率为94．74％。弹性义齿对Kennedy一类缺损修复的满意和比较满意率为66．67％；对Kennedy二类缺损修复的满意和比较满意率为50．00％；对Kennedy三类缺损修复的满意和比较满意率为96．15％；对Kennedy四类缺损修复的满意和比较满意率为84．21％。结论：(1)弹性义齿是一种美观、舒适、对基牙预备甚少的可摘局部义齿修复形式。（2）弹性义齿较适合咀嚼压力小而缺牙数目不多的前牙和前磨牙缺失的修复。     

吸氧对纤维支气管镜检查患者相关血气指标的干预和影响

目的:了解中等浓度吸氧在纤维支气管镜(FB)检查中的安全性和价值。方法:在行FB检查人群中随机选择25例，分别观察FB置人气道(置镜)前后及中等浓度吸氧前后患动脉血氧分压(PaO2)、动脉血二氧化碳分压(PaCO2)和血氧饱和度(SpO2)的变化。结果:置镜使患SpO2和Pa02有显下降(P＜0．001)，而PaCO2差异无显性(P＞0．05)。接着吸氧5L／min后，Sp02和PaO2与置镜后比较，有明显提高(P＜0．001)，而PaCO2变化无统计学意义(P＞0．05)。l3例患置镜前和吸氧后PaO2及SpO2比较，差异均具明显显性(P＜0．01)，而PaCO2无明显变化(P＞0.05)。结论:吸中等浓度氧对普通行FB检查的患是安全的，可完全补偿因FB置入所引起的PaO2下降，而对PaCO2影响很小，对于有原发PaCO2升高的患，安全性尚有待进一步研究。     

脂质体携载SOD的实验研究

本文采用去垢剂透析法成功制备出携载ＳＯＤ的脂质体（Ｌ－ＳＯＤ），其直径为１０２ｎｍ，对ＳＯＤ的包裹率为２１％，９０％以上酶活性存在于脂质体内部。将Ｌ－ＳＯＤ注射到大鼠静脉血中，其半衰期超过４ｈ，明显长于天然ＳＯＤ（８ｍｉｎ）。表明Ｌ－ＳＯＤ优于天然ＳＯＤ，具有临床应用前景。     

维生素E脂质体的制备

用超声波制备了维生素Ｅ脂质体，测定了脂质体的包封率及放置后的稳定性；观察了脂质体的形态，粒度分布。０．２％维生素Ｅ脂质体用于治疗老年性角化斑，试用２０例总有效率达９５％，优于１例维生素Ｅ乳剂。