纳米活性碳吸附表阿霉素的结构测定

目的通过测定纳米活性碳吸附的表阿霉素经脱吸附后,脱吸附出来的表阿霉素的结构是否发生改变,来推测纳米活性碳吸附表阿霉素的稳定性,进而为临床使用纳米活性碳吸附表阿霉素行局部淋巴化疗寻找实验依据。方法通过物理脱吸附方法使表阿霉素-纳米活性碳混悬液中纳米活性碳吸附的表阿霉素释放出来,再通过超高效液相色谱质谱联用法（LC-MS法）测定脱吸附后的表阿霉素结构是否发生改变。结果表阿霉素-纳米活性碳混悬液经过脱吸附后提取的表阿霉素行LC-MS法检测提示：脱吸附后提取的表阿霉素样品液与表阿霉素标准液的LC-MS图基本一致。结论被纳米活性碳吸附的表阿霉素,经脱吸附后其结构并未发生改变。     

表阿霉素纳米胶束靶向治疗大鼠移植性肝癌

目的 探讨键合表阿霉素纳米胶束(EPI-NPs)在大鼠移植性肝癌模型肝内的分布及药效.方法 制备具有靶向控释作用的EPI-NPs,Walker-256细胞直接注射法制作大鼠移植性肝癌模型,通过肝动脉灌注给药(EPI含量均为2 mg/kg),利用组织荧光和匀浆荧光技术对比分析EPI-NPs和表阿霉素(EPI)在肝内的分布特点,并作药效对比.结果 EPI-NPs组肿瘤组织的EPI荧光强度高于肝组织(P＜0.05),而EPI组则表现为平均分布;EPI-NPs组在肿瘤及肝内的EPI荧光强度均高于EPI组(P＜0.05).EPI-NPs组大鼠肝移植瘤的体积抑瘤率及质量抑瘤率分别为72.96％和68.19％,EPI组的体积抑瘤率及质量抑瘤率分别为39.16％和35.33％,两者差异均有统计学意义(P＜0.05).EPI-NPs组生存率高于EPI组(P＜0.05).结论 EPI-NPs具有良好的肿瘤组织靶向性和抑瘤效果.     

外磁场协同表阿霉素-羧甲基磁性纳米颗粒对膀胱癌体内外的杀伤作用

目的探讨脉冲外电磁场协同表阿霉素(EPI)-羧甲基葡聚糖氧化铁磁性纳米颗粒(CDMN)对人膀胱癌BIU-87细胞体外增殖活性和裸鼠皮下移植膀胱癌生长及其凋亡的影响。方法用化学共沉淀和氧化还原法制备EPI-CDMN并检测其理化性质,建立膀胱癌BIU-87细胞裸鼠皮下移植瘤动物模型,并设立空白对照组、磁场组、EPI组和EPI-CDMN组作为对照。分别采用噻唑蓝(MTT)比色法、双标流式细胞术、DNA原位末端标记法观察脉冲外电磁场联合EPI-CDMN对BIU-87细胞生长增殖活性和裸鼠皮下瘤生长状况及凋亡的影响。结果EPI-CDMN的直径、比饱和磁化强度分别为8～10 nm、0．22 emu/g。外磁场协同EPI-CDMN可以显著抑制BIU-87细胞增殖,抑制率达(21．82±3．18)%,并诱导细胞发生明显的凋亡,凋亡率为(16．8±3．4)%,均显著高于其他各组(P＜0．05)；裸鼠皮下瘤体积(1．57±0．42)cm~3和重量(2．00±0．21)g,均显著低于其他各组,抑瘤率51．69%和细胞凋亡指数(60．45±6．93)%均明显高于其他各组。结论外磁场可以显著增强EPI-CDMN对膀胱肿瘤的杀伤作用,为抗膀胱肿瘤的磁靶向治疗提供了实验依据。     

优化工艺制备表阿霉素α-聚氰基丙烯酸正丁酯纳米粒

目的以优化工艺制备表阿霉素α-聚氰基丙烯酸正丁酯纳米粒(EPI-PBCA-NP)。方法采用乳化聚合法制备PBCA-NP,再以二步法制备EPI-PBCA-NP,以形态、包封率和载药量为指标,应用U9(95)均匀试验优化处方工艺：右旋糖酐-70(Dextran-70)用量为10～90 mg,普流罗尼克F68(Pluronic F68)的用量为10～90 mg,表阿霉素的用量为1～7 mg；pH值的范围为1～5。结果制备EPI-PBCA-NP的优化条件为反应液pH为1,Pluronic F68 90．0 mg,Dextran-70 10．0 mg,表阿霉素为9．0 mg,在优化条件下制备EPI-PBCA-NP平均粒径为(135．5±16．8)nm(n=15)；平均包封率(86．78±11．20)%,平均载药量为(18．65±2．89)%。结论本研究通过优化工艺制得粒径满意的纳米粒,其包封率和载药量均高于单纯性表面吸附或包囊方式。     

纳米金对表阿霉素的增敏作用

目的 观察纳米金对表阿霉素在抑制HepG2细胞增殖中如何发挥增敏作用.方法 实验组分为纳米金预处理组和单纯表阿霉素组.常规消化HepG2细胞后种板,各组分别加入2 μg/L的纳米金溶液和无血清培养液100μl,于设定的时间点(30、60、120和240 min)加入1 mg/L的表阿霉素溶液100μl,继续培养24 h后噻唑蓝(MTT)比色法检测两组药物对HepG2细胞的增殖抑制作用;紫外-可见分光光度计(UV-Vis)检测各组HepG2细胞内积聚的表阿霉素量;原子力显微镜(AFM)观察HepG2细胞表面形态及超微结构变化.结果　纳米金预处理组各时间点HepG2细胞增殖抑制率[(50.53±1.38)%、(51.83±0.47)%、(48.66±2.21)%、(43.55±1.01)%]均明显高于对应的单纯表阿霉素组[(37.24±3.49)%、(39.42±2.28)%、(34.98±2.27)%、(28.92±3.80)%],P值＜0.01;纳米金预处理组(60min)表阿霉索在HepG2细胞内积聚的量最多,为(4.01±0.14)μg;AFM检测到纳米金预处理组HepG2细胞膜内陷,粗糙度增加,表面孔径增大,细胞核的饱满程度减少.结论　纳米金可通过增加表阿霉素在HepG2细胞内的积聚量来发挥增敏作用,纳米金预处理60 min对表阿霉素的增敏作用最大.

Abstract：

Objectiye To observe the sensitization of gold nanoparticles on the epirubicin therapy of hepatocellular carcinoma cells (HepG2) and action mechanism. Methods The HepG2 cells were divided into 2 experimental groups: gold nanoparticles pretreatment group and simple epirubicin group. After seeded in a 96-well plate and cultured for 24 h, HepG2 cells were treated with 100 μl of gold nanoparticles (2 μg/L) and serum-free medium respectively. Then 100 μl of epirubicin ( 1 mg/L) was added to both groups at different time points: 30, 60, 120 and 240 min. The inhibition effect of epirubicin on the HepG2cells was assessed by methyl thiazol tetrazolium (MTT) assay. Ultraviolet-visible spectrophotometer (UV-Vis) was applied to detect the epirubicin accumulation in HepG2 cells at different time points, and atomic force microscope (AFM) was used to examine the ultrastructural changes of HepG2 cell surface. Results The inhibition rate of HepG2 cells in gold nanoparticles pretreatment group [(50. 53 ± 1.38 ) %,(51.83 ± 0. 47 ) %, (48. 66 ± 2.21 ) %, (43.55 ± 1.01 ) %] at different time points was higher than the simple epirubicin group remarkably [( 37.24 ± 3.49 ) %, ( 39. 42 ± 2. 28 ) %, ( 34. 98 ± 2. 27 ) %,(28.92 ±3. 80)%] ,P ＜0. 01. The gold nanoparticles pretreatment group (60 rain) had the maximal epirubicin accumulation: (4. 01 ±0. 14) μg. AFM revealed that when treated with gold nanoparticles, the morphologic changes of HepG2 cells were significantly different: there appeared invagination, obvious shrinked cell membrane and much rougher surface. Conclusion Gold nanoparticles can sensitize epirubicin through the enhancement of epirubicin accumulation in HepG2 cells. The pretreatment of gold nanoparticles for 60 min has the maximum sensitization.     

表阿霉素诱导肝癌细胞凋亡的研究

目的 研究表阿霉素对肝癌细胞凋亡的影响.方法 在体外培养的肝癌细胞中加入不同浓度的表阿霉素,应用光镜、电镜、DNA凝胶电泳及流式细胞术检测肝癌细胞凋亡的形态学特征、生化学特征.细胞凋亡百分率及细胞周期的变化.结果 在表阿霉素作用下,凋亡的肝癌细胞出现膜小泡、凋亡小体等特征性改变;DNA电泳呈现典型的“梯状”条带;流式细胞术测定,出现典型的凋亡峰,其凋亡百分率随着药物浓度的提高和作用时间的延长而明显升高;同时,G_(1/0)期和S期细胞含量下降,而G_2/M期细胞含量上升.非凋亡细胞则无此表现.结论 表阿霉素可诱导肝癌细胞发生凋亡,并可使肝癌细胞生长受阻于G_2/M期,这可能是其杀伤肿瘤的一种重要机制.     

表阿霉素聚氰基丙烯酸正丁酯磁性纳米粒的制备与表征

目的 制备表阿霉素聚氰基丙烯酸正丁酯磁性纳米粒(EPI-PBCA-MNPs)并进行表征.方法 以化学共沉淀法制得粒径(15.2±4.6)am的Fe3O4纳米磁流体,采用微乳液反相界面聚合法制得EPI-PBCA-MNPs,反应条件:pH=2.0,搅拌速度2000 r/min,然后对其进行相关表征.结果 EPI-PBCA-MNPS粒径为(152.0 ±28.5)nm,分布均匀,胶体溶液长期稳定,平均包封率为(81.30 ±15.20)%,平均载药量为(14.65±2.05)%,饱和磁化强度为:0.35 KA/m.结论 制备的EPI-PBCA-MNPs粒径合适,分布均匀,其主要指标符合肝癌靶向治疗的要求.

Abstract：

Objective To prepare and characterize epirubicin polybutylcyanoacrylate magnetic nanoparticles ( EPI-PBCA-MNPs) for the purpose of offering a new dosage form of high-performance for targeted therapy of hepatoma. Methods Nanosized Fe3 O4 magnetic fluids with the mean particle diameter of (15. 2 ±4. 6) nm were prepared by chemical coprecipitation method. Subsequently, EPI-PBCA-MNPs was prepared by a reversed-phase micro-emulsion polymerization method, of which reactive condition was pH = 2. 0 and agitating velocity = 2000 r/min, and was correlatively characterized. Results EPI-PBCA-MNPs were uniformly and stably distributed in the colloid solution with the mean particle diameter of (152.0 ±28. 5) nm. The mean envelopment rate, drug-loading, saturated magnetization intensity is (81. 30 ±15. 20) % ,(14. 65 ± 2.05) % ,0. 35 KA/m, respectively. Conclusion In this experiment, EPI-PBCA-MNPs is uniformly distributed with satisfactory particle diameter,of which main indexes meet the demand of targeted therapy for hepatoma, offering a new dosage form with applicable potential for anticancer drugs exploration.     

索拉非尼联合表阿霉素对乳腺癌MCF-7细胞作用的研究

目的:探讨索拉非尼(Sorafenib)联合表阿霉素(Epirubicin)对乳腺癌MCF-7细胞的作用。方法:实验分为4组:空白对照组、索拉非尼单药组、表阿霉素单药组、索拉非尼联合表阿霉素组。根据索拉非尼及表阿霉素的IC50值设定联合给药浓度及时间。MTT法测定72h时间点索拉非尼及表阿霉素单用与联合对MCF-7细胞的增殖抑制率。结果:索拉非尼72h的IC50值为1.820μmol/l,表阿霉素72的IC50值为0.99μmol/l。在72h时,两药联合给药表现为协同或相加作用(P<0.05)。结论:索拉非尼和表阿霉素对乳腺癌细胞MCF-7细胞的增殖有抑制作用,联合给药表现出协同或相加作用。     

表阿霉素预防浅表性膀胱肿瘤术后复发的研究

目的研究表阿霉素预防浅表性膀胱肿瘤术后复发的可能机制。方法通过细胞培养、免疫组织化学研究不同浓度表阿霉素对膀胱肿瘤EJ细胞株的Ki-67的表达的影响，RT—PCR方法检测术前24h分别接受生理盐水、阿霉素、表阿霉素膀胱灌注患者膀胱肿瘤组织Fas—mRNA的表达。结果0、30、60、90mg／L表阿霉素各浓度组Ki-67的阳性率分别为79．60±3．50、60．73±3．31、51．57±3．93、45．83±3．75（P〈0．01）；表阿霉素较生理盐水可以明显促进肿瘤组织中Fas—mRNA的表达（P〈0．01）表阿霉素组和阿霉素组比较差异无统计学意义（P〈0．05）。结论表阿霉素可能通过下调Ki-67抑制膀胱肿瘤细胞增殖，促进Fas—mRNA的表达诱导凋亡达到抗肿瘤作用．     

纳米金增加阿霉素耐药肝癌细胞株敏感性的实验研究

目的：观察纳米金（GNP）人耐药肝癌细胞株耐药性的逆转作用。
　　方法：采用氯金酸柠檬酸三钠还原法制备并鉴定;分别用GNP与阿霉素（ADM）组单独或联合作用于ADM耐药的肝癌细胞株HepG2/ADM,以未处理的HepG2/ADM细胞为对照,用MTT法检、流式细胞术检测细胞的增殖与凋亡情况;紫外分光光度计检测HepG2/ADM细胞经ADM单独作用以及GNP与ADM联合作用后细胞内ADM浓度。
　　结果：与对照细胞比较,ADM单独作用及GNP与ADM联合作用后,HepG2/ADM的增殖均明显抑制、凋亡率明显升高,但后者的作用明显强于前者（均P<0.05）,而GNP单独作用对细胞的增殖与凋亡无明显影响（均P>0.05）;GNP+ADM作用后,HepG2/ADM细胞内的ADM含量较ADM单独作用后的ADM含量明显增加[（2.92±0.13）μg/Lvs.（1.68±0.74）μg/L,P<0.05]。
　　结论：GNP可增加HepG2/ADM细胞对ADM的敏感性,该作用可能与其增加HepG2/ADM细胞内的ADM浓度有关。     

纳米炭混悬液在非小细胞肺癌外科治疗中的临床应用

目前,外科治疗是非小细胞肺癌(non-small cell lung cancer,NSCLC)主要的治疗方法之一。手术治疗NSCLC的标准术式是切除原发病灶和清扫肺门及纵隔淋巴结。尽管如此,早期肺癌患者术后的复发率仍然很高。其原因主要为两个方面,一是清扫淋巴结不彻底;二是淋巴结微转移(micrometastasis,MM)。纳米炭混悬液的出现为我们解决上述问题提供了一种新的思路。纳米炭混悬液是一种淋巴示踪剂,其主要作用是淋巴示踪和靶向化疗。淋巴示踪使术中对淋巴结的清扫更彻底、更完全;靶向化疗可以将术中残留的肿瘤细胞进一步清除,以防止术后肿瘤复发。在胃肠道肿瘤和乳腺肿瘤的外科治疗中,纳米炭混悬液已得到较广泛的应用,作用和疗效肯定;而纳米炭混悬液在肺癌患者外科治疗中的应用是一种尝试,它可在术中给胸外科医生提供一定的帮助,使肺癌患者获得更好的预后。现对纳米炭混悬液的特点、应用方法、在NSCLC外科治疗中的临床应用进行综述。     

具有近红外荧光的二氧化钛纳米粒的载药与体外释放

目的 制备具有近红外荧光的纳米二氧化钛（TiO2）,并考察纳米粒的载药及体外释放性能。 方法 通过水热法合成掺杂钐的TiO2（Sm-TiO2）,采用透射电镜（TEM）对其进行表征,测定其荧光光谱,并考察对多柔比星（DOX）的载药量及体外释放曲线。结果 所制备的纳米粒分散均匀,外观呈梭状,长度100～200 nm,发射波长640～670 nm,在水中的载药量达11.5%,体外释放具有pH敏感性。结论 所制备Sm-TiO2有良好的近红外荧光发光效果、较高的载药量及可控的体外释放,可以作为新型药物载体深入研究。     

艾地苯醌纳米结构脂质载体的体外释放研究

目的：研究能够提高艾地苯醌生物利用度,并延长其体内循环时间的纳米脂质载体系统,运用透析袋扩散法,考察其体外释放特性。方法：通过研究不同形式的艾地苯醌载药方式（纳米结构脂质载体（NLC）、纳米乳（NE）及其水凝胶）在不同的环境（PH1.2、PH6.8、PH7.4）以及不同的摇床转速等变量情况下的体外释放特性,得以模拟在胃液、肠液和体液环境下,脂质纳米载体、水凝胶的缓释和控释性能。结果：艾地苯醌不同载药方式体外释放特性不同,NE剂型的释放速度较快24 h时约有22%的药物被释放;NLC剂型体外释放速率次之,在24 h时约有17%的药物被释放;自由药物体外释放速率最慢,在24 h时只有5%的药物被释放。水凝胶有明显的药物缓释作用;另外,在一定范围内,摇床转速越高,药物的缓释效果更明显。结论：艾地苯醌纳米结构脂质载体的体外释放研究方法简单、快速,可以用于用以控制制剂质量,评价制剂工艺。     

Basophil Histamine Release in Asthma Patients after in Vitro Provocation with Althesin and Etomidate

Our aim was to compare the histamine-releasing effect of etomidate and Althesin on basophil leukocytes from asthmatic patients and normal persons. Blood from eight asthmatic patients and six normal persons was tested for histamine release after in vitro provocation with etomidate and althesin. In the group of asthmatic patients there was a significantly higher histamine release after provocation with althesin than after provocation with etomidate at all concentrations (P less than 0.05, P less than 0.01, P less than 0.02). There was significantly higher histamine release for asthmatic patients than for normal persons after provocation with althesin at all concentrations (P less than 0.05, P less than 0.01, P less than 0.02). There was no difference between the asthma group and the normal group after provocation with etomidate. Data were analysed using Wilcoxon's and Mann-Whitney's rank sum tests. We conclude that asthmatic patients may risk bronchospasm during induction of anaesthesia with althesin, and that etomidate may be suitable intravenous anaesthetic for asthmatic patients.     

Investigation of Plasma Protein Adsorption on Functionalized Nanoparticles for Application in Apheresis

Particles with specific ligands for the adsorption of plasma proteins can be used in therapeutic or preparative apheresis. The development of these particles may benefit from an improved knowledge of the relationship between protein adsorption and the structure of ligands. Nanoparticles were functionalized with aliphatic diamines of increasing chain length; with the amino acids lysine, tryptophan, histidine, and their corresponding amines; and with tryptophan and histidine spaced with diamines of different length. Suitable protocols were developed for the washing of particles and the subsequent desorption of proteins adsorbed from human plasma. The adsorption pattern, as well as the quantification of the overall adsorption of proteins on these modified particles, was investigated with gel electrophoresis. This was followed by immuno-blotting which yielded specific assessments of bound human serum albumin and fibrinogen. The comparison of protein adsorption with surface charge density and measured hydrophobicities yielded no simple correlations although in general more hydrophobic ligands bound higher quantities of protein. The detection of human serum albumin yielded similar results because it was observed for overall protein adsorption while the adsorption of fibrinogen expressed a different pattern. In this case, particular nanoparticles functionalized with aliphatic diamines bound significantly higher amounts of fibrinogen than all other ligands.     

肾康注射液拮抗马兜铃酸对人近端肾小管上皮细胞作用的研究

目的：探讨肾康注射液（SKI）能否拮抗马兜铃酸钠盐（AA-Na）诱发的人近端肾小管上皮细胞（HKC）的促纤维化效应。方法：AA-Na（10mg/L）加或不加SKI（8mg/ml）与HKC孵育,然后检测转化生长因子-β1（TGF-β1）、结缔组织生长因子（CTGF）、金属蛋白酶组织抑制物-1（TIMP-1）及纤溶酶原激活物抑制物-1（PAI-1）的mRNA表达（孵育12h,RT-PCR方法检测）和蛋白质表达（孵育36h用免疫印迹法检测胞内CTGF,孵育24h用ELISA法检测上清中其他因子）。结果：AA-Na能显著上调HKC对TGF-β1、CTGF、TIMP-1、PAI-1的表达,与对照组比较,mRNA表达分别上调1.84,1.58,1.62,1.29倍,蛋白质表达分别上调1.12,1.63,1.42,1.29倍,P均〈0.05;加SKI后,上述因子的高表达均被显著抑制,与AA-Na组比较,mRNA表达的抑制率分别为41.6%,43.7%,43.8%及24.4%,蛋白质表达的抑制率分别为34.3%,43.3%,31.1%及21.9%,P均〈0.05。结论：AA-Na能刺激HKC显著上调促细胞外基质（ECM）合成因子（TGF-β1、CT-GF）及抗ECM降解因子（TIMP-1、PAI-1）的mRNA及蛋白质表达,而SKI能拮抗AA-Na的上述作用。     

Choice of Corticosteroid Solution and Outcome After Injection for Trigger Finger

Background: Many techniques for injection of trigger fingers exist. The purpose of this study was to determine whether the type of steroid or technique used for trigger finger injection altered clinical outcomes. Methods: Six hand surgeons at a single institution were surveyed regarding their injection technique, preferred steroid used, and protocol for repeat injection or indication for surgery for symptomatic trigger finger. A retrospective chart review of patients who underwent trigger finger injections was performed by randomly selecting 35 patients for each surgeon between January 2013 and December 2015. Demographic data at the time of presentation were collected. Outcome data during follow-up appointments were also recorded. Results: A total of 210 patient charts were reviewed. Demographic data and initial presenting grade of triggering were similar among all groups. There was no significant difference in clinical course or eventual outcomes noted with injection technique. There were 70 patients in each steroid cohort. Patients receiving triamcinolone required additional injections compared with those receiving methylprednisolone and dexamethasone. Eventual surgical intervention was significantly higher in those patients receiving methylprednisolone. The methylprednisolone group also underwent operative release significantly earlier. Conclusions: Trigger finger injections with triamcinolone demonstrate a higher rate of additional injections when compared with dexamethasone and methylprednisolone. Patients who underwent methylprednisolone injection had surgical release performed earlier and more frequently than the other 2 groups. The choice of corticosteroid significantly affected clinical outcome in this study population. Clinicians performing steroid injections for trigger finger may wish to consider these results when selecting a specific agent.     

载顺铂的纳米羟基磷灰石对A549肺腺癌细胞体外胀亡的影响

目的探讨载顺铂的纳米羟基磷灰石对体外培养的人肺腺癌细胞A549增殖及胀亡的影响。方法用机械融合法制备载顺铂的纳米羟基磷灰石粒子,将肺腺癌A549细胞暴露于该粒子中培养。进行形态学观察,MTT法检测载顺铂的纳米羟基磷灰石粒子体外作用于人肺癌A549细胞的量效和时效关系,流式细胞仪分析细胞周期的时相变化。结果载顺铂的纳米羟基磷灰石粒子在体外对人肺癌A549细胞系具有明显的抑制作用,并呈现良好的量效和时效关系。肺癌细胞的生长阻滞于G2/M期,细胞体积增大,胞质空泡化,染色质凝集,胞浆溶解。结论载顺铂的纳米羟基磷灰石粒子具有体外抗肺腺癌系A549的作用,细胞增殖阻滞于G2/M期,阻断细胞周期的进展,导致癌细胞溶解坏死,诱导细胞凋亡是其可能的机制之一。