柔红霉素毫微粒冻干针剂的研究

目的：制备易再分散、稳定的柔红霉素聚氰基丙烯酸正丁酯毫微粒（ＤＮＲ－ＰＢＣＡ－ＮＰ）冻干针剂。方法：选用适宜支架剂制得ＤＮＲ－ＰＢＣＡ－ＮＰ冻干针剂,并评价其相关理化性质。结果：冻干前后毫微粒形态、粒径、ｐＨ、包封率及载药量均无明显变化,含水量合格,再分散性良好,制剂稳定。其临界相对湿度为７５．３３％。结论：在适宜的处方及工艺条件下制备ＤＮＲ－ＰＢＣＡ－ＮＰ冻干针剂是可行的。     

米托蒽醌聚乳酸缓释毫微粒冻干针剂在动物体内的靶向性研究

目的：比较肝靶向米托蒽醌聚乳酸缓释毫微粒（ＤＨＡＱ－ＰＬＡ－ＮＰ）冻干针剂和ＤＨＡＱ水针剂在小鼠体内的分布规律,验证前者的肝靶向性。方法：采用ＨＰＬＣ法测定静注ＤＨＡＱ－ＰＬＡ－ＮＰ和ＤＨＡＱ水针剂后小鼠血液、心、肝、脾、肺、肾的药物浓度,由此计算各器官的相对百分含量。结果：ＤＨＡＱ－ＰＬＡ－ＮＰ冻干针剂在肝脏的分布明显高于ＤＨＡＱ水针剂,在其它器官中的含量则低于水针剂,给药２４小时后药物在肝中的     

阿柔比星A聚乳酸毫微粒冻干针剂的研制及体外药物释放规律的研究

研究阿柔比星Ａ聚乳酸毫微粒（ＡＣＲＢ－Ａ－ＰＬＡ－ＮＰ）冻干针剂的制备,并对其体外释药进行考察。根据低共熔点测定结果,以及外观、色泽、再分散性等质量参数为指标,制备出ＡＣＲＢ－Ａ－ＰＬＡ－ＮＰ冻干针剂,采用动脉透析法研究其体个释放规律。结果,制得的ＡＣＲＢ－ＡＰＬＡ－ＮＰ冻干针剂色泽均匀,再分散性良好。其体外释药可用３种方式表达。提示ＡＣＲＢ－Ａ－ＰＬＡ－ＮＰ冻干针剂有明显的缓释特征。     

阿柔比星A聚乳酸毫微粒冻干针剂小鼠体内分布研究

对阿柔比星Ａ冻干针剂与阿柔比星Ａ聚乳酸毫微粒冻干针剂给予小鼠尾静脉注射后体内分布比较，采用ＨＯＬＣ法分别测定小鼠给药后血液及主要脏器中ＡＣＲＢ０Ａ浓度。结果阿柔比星Ａ聚乳酸毫微粒在小鼠肝脏中的浓度高于对照品浓度。     

Velvet antler polypeptides promoted proliferation of chondrocytes and osteoblast precursors and fracture healing

目的：研究鹿茸总多肽（ＶＡＴＰ）和鹿茸多肽ＶＡＰＡ,ＶＡＰＢ和ＶＡＰＣ对细胞增殖的影响．方法：分离家兔和人胚软骨细胞及鸡胚头盖骨细胞,体外加入ＶＡＴＰ,ＶＡＰＡ,ＶＡＰＢ和ＶＡＰＣ,观察其对［３Ｈ］ＴｄＲ参入细胞ＤＮＡ合成的影响．整体观察了ＶＡＴＰ促进实验性大鼠桡骨骨折愈合作用．结果：ＶＡＴＰ５０－２００ｍｇ·Ｌ－１和ＶＡＰＢ１２５－５０ｍｇ·Ｌ－１促进软骨及成骨样细胞有丝分裂,ＶＡＰＡ作用较弱,ＶＡＰＣ无效．ＶＡＴＰ１０－２０ｍｇ·ｋｇ－１加速了大鼠桡骨骨折的愈合．结论：ＶＡＴＰ加速骨折愈合的作用与其促进软骨和成骨样细胞的增殖有关     

放射性碘标记毫微粒给小鼠灌胃后的示踪动力学实验研究

采用氯氨－Ｔ法用＾１２５Ｉ标记ＢＳＡ，再次其掺入到制备毫微粒的材料中，制备成放射标记的毫微粒，粒径在１５０－２１０ｎｍ之间。我们通过核素示踪法和放射自显影，对（两种）毫微粒给小鼠灌胃后的吸收，分布及清除进行研究。     

阿柔比星A聚乳酸毫微粒主要性质研究

目的：对载药毫微粒主要质量指标载药量、包封率及其关系,粒径及其分布进行研究,方法：以阿柔比星Ａ聚乳酸微粒为研究对象,以分光光度测定载药量与包封率,以激光粒度分析仪测定粒径及其分布。结果：阿柔比星Ａ聚乳酸毫微粒平均载药量为１８．５％。平均包封率为８６．７％,平均数目径为８０ｎｍ,平均体积径为２３０ｎｍ。结论：载药量与包封率之间具有一定关系。体积径分布是载药毫微粒粒径分布评价不可忽视的内容。     

阿克拉霉素A毫微囊在大鼠体内的组织分布研究

本文利用反相高效液相色谱法测定了阿克拉霉素Ａ毫微囊（ＡＣＭ－Ａ－ＮＣ）在大鼠体内各组织的分布，结果表明：阿克拉霉素Ａ在肺，脾，小肠和胸腺分布较多，而心，肝、肾中分布较少，与ＡＣＭ－Ａ水溶液对比研究表明：ＡＣＭ－Ａ－ＮＣ在肺内分布量较其水溶液提高３．６倍。在脾中提高１．４位，肝中２．２倍，肾中１．６倍，胸脾中１．２倍，大肠１．１倍，心中约１．２倍。     

α－常春藤皂甙和无患子皂甙B对小鼠肝微粒体细胞色素P－450的作用

目的：研究α常春藤皂甙（Ｈｅｄ）和无患子皂甙Ｂ（ＳａｐＢ）对小鼠肝细胞色素Ｐ４５０的影响与保肝作用的关系．方法：给小鼠ｓｃＨｅｄ，ＳａｐＢ和Ｈｅｄ＋ＳａｐＢ（１∶１５），检测小鼠肝微粒体细胞色素Ｐ４５０的含量．结果：Ｈｅｄ２０ｍｇ·ｋｇ－１，ＳａｐＢ２０ｍｇ·ｋｇ－１和Ｈｅｄ＋ＳａｐＢ２０ｍｇ·ｋｇ－１使肝细胞色素Ｐ４５０分别降低了４０％，５５％和５０％．这种抑制作用在３ｄ后基本恢复正常．苯巴比妥ｉｐ５０ｍｇ·ｋｇ－１使小鼠肝细胞色素Ｐ４５０增加２５倍，Ｈｅｄ＋ＳａｐＢ使苯巴比妥诱导的Ｐ４５０降低了５０％，小鼠肝微粒体体外与Ｈｅｄ＋ＳａｐＢ共孵对Ｐ４５０没有影响．结论：Ｈｅｄ和ＳａｐＢ的保肝作用至少在某一方面是由于降低了肝细胞色素Ｐ４５０而产生的．     

阿克拉霉素A毫微囊在大鼠体内的组织分布研究

本文利用反相高效液相色谱法测定了阿克拉霉素Ａ毫微囊（ＡＣＭ－Ａ－ＮＣ）在大鼠体内各组织的分布．结果表明：阿克拉霉素Ａ在肺、脾、小肠和胸腺分布较多，而心、肝、肾中分布较少．与ＡＣＭ－Ａ水溶液对比研究表明：ＡＣＭ－Ａ－ＮＣ在肺内分布量较其水溶液提高３．６倍，在脾中提高１．４倍，肝中２．２倍，肾中１．６倍，胸脾中１．２倍，大肠１．２倍，小肠１．１倍，心中约１．２倍．     

Indirect Evidence that Drug Brain Targeting Using Polysorbate 80-Coated Polybutylcyanoacrylate Nanoparticles Is Related to Toxicity

Purpose. To investigate the mechanism underlying the entry of the analgesic peptide dalargin into brain using biodegradable polybutylcyanoacrylate (PBCA) nanoparticles (NP) overcoated with polysorbate 80. Methods. The investigations were carried out with PBCA NP and with non biodegradable polystyrene (PS) NP (200 nm diameter). Dalargin adsorption was assessed by HPLC. Its entry into the CNS in mice was evaluated using the tail-flick procedure. Locomotor activity measurements were performed to compare NP toxicities. BBB permeabilization by PBCA NP was studied in vitro using a coculture of bovine brain capillary endothelial cells and rat astrocytes. Results. Dalargin loading was 11.7 µg/mg on PBCA NP and 16.5µg/ mg on PS NP. Adding polysorbate 80 to NP led to a complete desorption. Nevertheless, dalargin associated with PBCA NP and polysorbate 80 induced a potent and prolonged analgesia, which could not be obtained using PS NP in place of PBCA NP. Locomotor activity dramatically decreased in mice dosed with PBCA NP, but not with PS NP. PBCA NP also caused occasional mortality. In vitro, PBCA NP (10 µg/ml) induced a permeabilization of the BBB model. Conclusions. A non specific permeabilization of the BBB, probably related to the toxicity of the carrier, may account for the CNS penetration of dalargin associated with PBCA NP and polysorbate 80.     

Preparation of carboxy‐PEG‐PLA nanoparticles loaded with camptothecin and their body distribution in solid tumor–bearing mice

Carboxy‐polyethylene glycol‐poly(DL‐lactic acid) block copolymer was synthesized by reductive amination of carboxy‐polyethylene glycol‐amine and aldehyde‐ended poly(DL‐lactic acid). The obtained product, PA‐PEG‐CB, composed of pure carboxy‐polyethylene glycol‐poly(DL‐lactic acid) block copolymer and poly(DL‐lactic acid), was used to prepare nanoparticles loaded with camptothecin (CPT), designated CPT‐NP. CPT‐NP with 500–600 nm and approximately 1% (w/w) drug content were obtained by emulsification/evaporation, while nanoparticles with a drug content of only less than 0.1% (w/w) were prepared by dialysis. CPT‐NP produced by emulsification/evaporation were used for in vitro and in vivo studies. CPT‐NP released CPT gradually in phosphate‐buffered saline, pH 7.4, at 37°C over 48 h. Biodistribution profiles were compared between CPT‐NP and CPT solution after i.v. injection into mice bearing sarcoma 180 solid tumor. CPT‐NP retained CPT approximately 10 times more in plasma during the initial 8 h than CPT solution. The tumor level of CPT was more than 10 times higher with CPT‐NP for the observation period (24 h), though the CPT concentration in the liver and spleen was much higher with CPT‐NP. CPT‐NP may be useful for tumor localization of CPT. Drug Dev Res 70: 512–519, 2009. © 2009 Wiley‐Liss, Inc.     

Galactose-modified cationic liposomes as a liver-targeting delivery system for small interfering RNA

We have developed a galactose-modified cationic liposome for delivery of small interfering RNA (siRNA) to the liver. The liposomes were designed to be transported into hepatocytes via the asialoglycoprotein receptor, which recognizes galactose residues. The liposomes contained a novel galactose-modified lipid, 1,2-dioleoyl-sn-glycerol-3-phosphatidyl-N-(1-deoxylactito-1-yl)ethanolamine (GDOPE). Delivery of siRNA to hepatocytes by the liposomes was evaluated by measuring the gene-silencing activity of liposome : siRNA complexes in two human hepatoma cell lines. A formulation with a cationic lipid : GDOPE ratio of 3 : 5 by weight, LIC-G5, showed the strongest activity. In mice, intravenous injection of LIC-G5 complexed with (3)H-labeled siRNA led to accumulation of radioactivity in the liver. When the hepatic cellular uptake was determined after intravenous injection into mice followed by collagenase liver perfusion, the distribution of siRNA to parenchymal cells was 1.9 times higher when LIC-G5 rather than nongalactosylated LIC was used as the carrier. The concentration of siRNA accumulated was 45 μg/ml, 30 times the concentration that produced strong gene silencing in vitro and therefore presumably sufficient for a therapeutic effect. Because increasing the cationic-lipid content of a liposome carrier generally enhances the uptake of siRNA by the liver at the expense of increased cell toxicity, we used only a moderate amount of cationic lipid in our galactose-modified carrier. LIC-G5 enhanced the uptake of siRNA by the liver without cytotoxic effects and is a promising candidate delivery system for liver-targeted siRNA therapy.     

眼用诺氟沙星毫微粒制备工艺的初选及优化

目的:寻找眼用诺氟沙星毫微粒最佳的处方和制备工艺。方法:选择氰基丙烯酸正丁酯为载体材料,用乳化聚合法制备NFLXPBCANP,通过单因素试验初选、均匀设计优化处方和制备工艺。结果:在优化条件下制备的NFLXPBCANP为乳白色胶体溶液,毫微粒外形圆整光滑、分布均匀、不粘连,平均粒径553nm,包封率90.14%,载药量22.70%。结论:该优化条件可作为眼用诺氟沙星毫微粒最佳的处方和制备工艺。     

Physicochemical characterisation of cationic polybutylcyanoacrylate-nanoparticles by fluorescence correlation spectroscopy.

The aim of this study was to compare different physical and chemical methods with fluorescence correlation spectroscopy (FCS) in order to characterise cationic acrylate nanoparticles (NP), which can deliver oligonucleotides (ON) into mammalian cells. These positively charged nanoparticles were prepared from diethylaminoethyl dextran (DEAE-dextran) and poly(n-butyl-2-cyanoacrylate) (PBCA). NP consists of PBCA oligochains with an average size of PBCA 9 mer and were formed by entrapping DEAE-dextran and dextran 70,000 in high amounts into the particle matrix. The oligochain length of PBCA was investigated by mass-spectroscopy (MALDI TOF). The molecular weight of a particle with d = 108 nm was estimated to be approximately 3.6 x 10(8) Da. The mean size of the nanoparticles were in a range of dh = 130-140 nm, as determined independently by FCS and dynamic light scattering. Atomic force microscopy and scanning electron microscopy images confirm this size range. Furthermore, the particle mass of the PBCA-NP was estimated by FCS measurements. For this approach two new methods for fluorescence labelling of cationic particles were developed. Fluorescent labelled dextran 70,000 was entrapped into the particle matrix; in addition, the derivatisation of hydroxyl groups of the NP was achieved with 5-([4,6-dichlorotriazin-2-yl]amino) fluorescein (DTAF). ON can be localised in a complex with the NP by dual-colour fluorescence cross correlation spectroscopy measurements. The zetapotential of the unloaded NP was positively charged with about +39 mV and decreased down to -40 mV on addition of excess ON. After centrifugation quantification of the ON loading onto the particles by strong anion exchange high performance liquid chromatography (SAX HPLC) and FCS showed that approximately 20 microg ON per 100 g NP was adsorbed. The FCS measurements of the ON adsorption in situ was found to be much higher with approximately 95 microg ON per 100 g NP.     

骨髓靶向柔红霉素毫微粒的研究

采用乳液聚合法制备了柔红霉素聚氰基丙烯酸正丁酯毫微粒（１）,并对其形态,粒径分布,载药性,动物体内经时变化过程及骨髓靶向性进行了研究。结果表明１平均微粒径为７０ｎｍ,分布范围３０－２２０ｎｍ,包封率为９７．０％,载药一达５５．７％;小鼠胸脉注射相同剂量１及柔红霉素后,前者股骨内峰浓度提高到１．６２倍,ＡＵＣ提高到４．８７倍,总靶向效率从５．１７％提高到２４．１９％,表明１具骨髓靶向性。     

Lung surfactant DPPG phospholipid inhibits vaccinia virus infection

Vaccinia virus (VACV) was used as a surrogate of Variola virus (genus Orthopoxvirus), the causative agent of smallpox, to study orthopoxvirus infection via the respiratory airway. Lung surfactant, a physiological barrier to infection encountered by the virus, is predominantly composed of phospholipids whose role during orthopoxvirus infection has not been investigated. An attenuated Lister strain, derived from the traditional smallpox vaccine and the Western Reserve (WR) strain, lethal for mice infected by the respiratory route, were examined for their ability to bind various surfactant phospholipids. Dipalmitoyl phosphatidylglycerol (DPPG) was found to interact with both VACV strains. DPPG incorporated in small unilamellar vesicle (SUV-DPPG) inhibited VACV cell infection, unlike other phospholipids tested. Both pre-incubation of virus with SUV-DPPG and pretreatment of the cell with SUV-DPPG inhibited cell infection. This specific DPPG effect was shown to be concentration and time dependent and to prevent the first step of the viral cycle, i.e. virus cell attachment. Cryo-electron microscopy highlighted the interaction between the virus and SUV-DPPG. In the presence of the phospholipid, virus particles displayed a hedgehog-like appearance due to the attachment of lipid vesicles. Mice infected intranasally with VACV-WR pre-incubated with SUV-DPPG survived a lethal infection. These data suggest that DPPG in lung surfactant could reduce the amount of orthopoxvirus particles able to infect pneumocytes at the beginning of a respiratory poxvirus infection. The knowledge acquired during this study of virus-DPPG interactions may be used to develop novel chemotherapeutic strategies for smallpox.     

肝靶向万乃洛韦毫微粒的研究

为提高核苷类药物对乙型肝炎的疗效并降低其毒性,用乳化聚合法制备了万乃洛韦聚氰基丙烯酸正丁酯毫微粒,对其形态、大小及其分布、体外释药特性、载药量、初步稳定性、动物体内的分布和体外肝细胞的摄取情况进行了研究。结果表明,该毫微粒粒径d_av=104.77±11.78nm;载药量11.20%;包封率84.85%;体外释药符合双相动力学规律;对肝细胞具有通透性;静注后15min有74.49%集中在肝脏。提示,万乃洛韦毫微粒对于提高万乃洛韦对病毒性乙型肝炎的治疗效果和降低其对肾脏的毒性有意义。     

免疫抑制剂干扰联合瘤细胞接种构建肝癌移植瘤小鼠模型

目的：探讨构建肝癌移植瘤动物模型的最佳方法。方法选用BALB/c小鼠160只,随机分为地塞米松组、肝癌细胞注射组、地塞米松加肝癌细胞注射组及空白对照组,每组40只,雌、雄各半。地塞米松组在小鼠腹腔内注射地塞米松;肝癌细胞注射组于不同时间在肝脏直接接种H22肝癌细胞株3次;地塞米松加肝癌细胞注射组将上述2种方法联合使用;空白对照组注射生理盐水。1个月后,颈椎脱臼处死小鼠,取其肝组织,计算肝指数,苏木精-伊红染色（HE）后观察肝组织病理变化。结果地塞米松组、空白对照组未发现肝组织癌变,肝癌细胞注射组、地塞米松加肝癌细胞注射组癌变的百分比分别为71.4%（20/28）、91.2%（21/23）。结论肝癌细胞直接接种小鼠肝脏及联合地塞米松干扰均可建立肝癌移植瘤动物模型,但联合地塞米松干扰建模的成功率明显升高。     

Synthesis of high loading and encapsulation efficient paclitaxel-loaded poly(n-butyl cyanoacrylate) nanoparticles via miniemulsion

The manufacture of stable paclitaxel-loaded poly(n-butyl cyanoacrylate) (PBCA) nanoparticles containing high loading and encapsulation efficiency simultaneously were achieved in the presence of pluronic F127 via miniemulsion. It was found that both drug loading and encapsulation efficiencies of PBCA nanoparticles prepared by miniemulsion were higher (approximately three times) than those obtained by emulsion with similar paclitaxel content in the feed monomer (1%, w/w). Furthermore, the loading and encapsulation efficiencies increased concurrently (to a maximum of 4 and 80%, respectively) with increasing paclitaxel content and these nanoparticles were spherical in shape and with size near 100 nm. XRD patterns revealed that paclitaxel in particles was distributed in the molecular or amorphous state or in the form of small crystals. The in vitro drug release profile of drug-loaded PBCA nanoparticles prepared from miniemulsion exhibited a gradual release; more than 80% (w/w) of the paclitaxel was released after 96 h.