HPLC测定兔血浆及淋巴组织中的阿霉素

 目的：测定兔血浆及微量淋巴組织中阿霉素的含量，揭示阿霉素脂质体的靶向定位及生物体内的药物代谢 动力学特性。方法：取样量，血浆为0.5ml，淋巴組织为0.1g，加入磷酸盐緩冲液1ml(0.2mol/L，pH7.8)于冰浴 中超声匀浆，以盐酸柔红霉素为内标，采用有机相[氯仿-甲醇（4 ：1)]一步提取，40℃下氮气吹干，残渣用甲醇溶解， 进样75μl色谱条件为YWG-C18柱（150mmΧΦ4.6mm，粒径为10μm);流动相为甲醇-L腈-0.01mol/L磷酸二氢铵-冰醋酸（50：22：28：0.5);流速为1.2ml/min，激发波长为479nm，发射波长为587nm,检測器灵敏度为 H;室温下操作。结果：本方法线性范围：血浆阿霉素为10-1000ng/ml;淋巴组织阿霉素为0.05-5.0μg/g，两者 的线性相关系数r=0.9999(n=6);日内及日间重现性：RSD%在2.10-6.15之间；药物自血浆及淋巴组织的平均 回收率分别为100.11%和100.5%;灵敏度：血浆及淋巴组织中最低检測浓度分别为2ng/ml和0.025μg/g;方法 简便快速，一个样本从提取处理到色谱分离出结果，可在30min内完成。结论：本方法能够簡便、准确地測出兔血浆 及微量淋巴组织中阿霉素的含量，适于淋巴给药后阿霉素生物体内药物代谢动力学的研究。     

Marginal analysis applied to the dose-response curve

To improve on drug dosage decisions, the method of “marginal analysis,” traditionally found in the economics literature, is applied to the dose-response curve. Marginal analysis is first defined and explained, next the technique is redefined for the clinical setting of dose determination subject to drug efficacy and side effects, and finally an example using adjuvant adriamycin chemotherapy in osteo sarcoma is presented. By not using “marginal analysis,” clinicians may not be maximizing overall therapeutic success or patient survival. Implications for future clinical trials are discussed, and a suggestion for estimating dose-response is proposed. The technique described should be readily applicable to many dosing problems, and need not be restricted to chemotherapy dosing decisions.     

Self-assembled hydrogel nanoparticles responsive to tumor extracellular pH from pullulan derivative/sulfonamide conjugate: characterization,aggregation, and adriamycin release in vitro

Purpose. To investigate some physicochemical properties of self-assembled hydrogel nanoparticles of pullulan acetate (PA) and sulfonamide conjugates, as a potential tumor targeting drug carrier responsive to tumor extracellular pH. Methods. A new class of pH-responsive polymers was synthesized by conjugating a sulfonamide, sulfadimethoxine (SDM), to succinylated pullulan acetate (coohPA). The polymers formed self-assembled PA/SDM hydrogel nanoparticles in aqueous media, which was confirmed by fluorometry and field emission-scanning electron microscopy. The pH-dependent behavior of the nanoparticles was examined by measuring transmittance, particle size and zeta potential. Adriamycin (ADR) was tested for loading into and release from the nanoparticles at various pHs. Results. The mean diameters of all PA/SDM nanoparticles tested were <70 nm, with a unimodal size distribution. The critical aggregation concentrations at pH 9.0 were as low as 3.16 g/mL. The nanoparticles showed good stability at pH 7.4, but shrank and aggregated below pH 7.0. The ADR release rate from the PA/SDM nanoparticles was pH-dependent around physiological pH and significantly enhanced below a pH of 6.8. Conclusions. The pH-responsive PA/SDM nanoparticles may provide some advantages for targeted anti-cancer drug delivery due to the particle aggregation and enhanced drug release rates at tumor pH.     

Long-term follow-up of a phase II trial studying a weekly doxorubicin-based multiple drug adjuvant therapy for stage II node-positive carcinoma of the breast

Background. Combination chemotherapy improves outcomes in women with breast cancer (BC) that involves axillary nodes. This single-arm study aimed to evaluate the effectiveness of an intensive doxorubicin-based multi-drug regimen as adjuvant therapy in women with stage II, node positive breast cancer. Patients and methods. Between 7/80 and 8/85, 654 women, aged 25–73, who had a mastectomy for stage IIB BC were accrued. Patients with prior RT, chemotherapy, or surgical or radiation castration within 1 year of diagnosis were excluded. Treatment consisted of: 6 weekly courses of IV cyclophosphamide (C) 400 mg/m², doxorubicin (A) 10 mg/m², vincristine (V) 1 mg/m², fluorouracil (F) 400 mg/m², and a tapering course of prednisone followed by 12 courses of C 400 mg/m², A 20 mg/m², V 1 mg/m², F 400 mg/m² given every 2 weeks. Patients with estrogen receptor positive tumors received Tamoxifen 10 mg bid between weeks 8 and 30. Treatment did not exceed 8 months. Median follow-up is 13.1 years. Results. Six hundred thirty six patients are eligible. Fewer positive (+) nodes, premenopausal status, and positive progesterone receptor status are significantly (p < 0.05) associated with longer survival. At 10 years, 61% were relapse-free in the 1–3 +node group compared to 37 and 21% in the 4–9 and ≥10 +node groups, respectively (p = 0.0001). Relapse-free survival at 10 years is 50% for premenopausal and 45% for postmenopausal patients. Severe or life-threatening hematological toxicity was seen in 6/630 (<1%) patients. Four patients had severe (grade 3) neurotoxicity which resolved. No cardiac toxicity was observed. Conclusion. This adjuvant regimen compares favorably to other published adjuvant treatments with similar length of follow-up.     

bak基因过表达对阿霉素杀伤诱导EJ细胞凋亡的增敏作用

目的目的：探讨bak基因过度表达在膀胱癌EJ细胞凋亡途径中的作用以及对阿霉素的可能的增敏作用，并研究其作用机制。方法用脂质体法，膀胱癌细胞中转入bak／pcDNA3的设为实验组，空转pcDNA3的设为对照组，用原位杂交法检测bak在癌细胞中的表达，同时用阿霉素处理后，利用MTT法测出细胞生存率，进而用荧光分光光度计检测细胞内阿霉素的聚集量。比色法检测癌细胞内Caspase-3活性改变。结果脂质体法转基因后，bak的mRNA呈阳性表达，细胞生存率较对照组明显降低（P〈O．05）。癌细胞Caspase-3活性增强（P〈O．05），细胞内阿霉素聚集量元明显增加（P〉O．05）。结论bak基因对阿霉素杀伤膀胱癌细胞具有增敏作用，但并非通过帮助阿霉索进入细胞发挥作用，其作用机制与激活Caspase-3有关。     

复合抗氧化剂对大鼠肝癌阿霉素化疗外周血的保护作用

目的观察大鼠肝癌模型腹腔阿霉素化疗引起的外周血的变化以及复合抗氧化剂的保护作用。方法采用Walker-256肿瘤细胞株接种大鼠皮下。得到实体瘤，将实体瘤分割成小块植入大鼠肝脏，制成大鼠移植性肝癌模型，将肝癌模型大鼠随机分成3组，分别为模型组、单纯化疗组和抗氧化剂保护组，同时选取经手术但未接种肿瘤的大鼠作为阴性对照组。抗氧化剂保护组每日给予复合抗氧化剂灌胃，单纯化疗组和抗氧化剂保护组大鼠每2天进行1次腹腔阿霉素化疗。剂量为2mg／kg体重，第8周末处死大鼠，取抗凝全血测定外周血各项指标。结果与阴性组和模型组相比较。单纯化疗组的白细胞数（WBC）、淋巴细胞百分比（LYM％）、淋巴细胞数（LYM）、红细胞数（RBC）、红细胞压积（HCT）、血红蛋白（HGB）、平均血小板体积（MPV）显著降低（P〈0．05）。单核细胞百分比（MON％）、中性粒细胞百分比（GRA％）、单核细胞数（MON）、中性粒细胞数（GRA）、平均红细胞体积（MCV）、红细胞分布活力（RDW）、平均红细胞血红蛋白含量（MCH）、血小板数（PLT）、血小板分布活力（PDW）显著升高，而抗氧化剂保护组与单纯化疗组相比呈现出了明显的拮抗保护作用。结论大鼠肝癌腹腔阿霉素化疗可以造成外周血损伤，引起血象的明显改变。而复合抗氧化剂可以起到有效地保护作用。     

阿霉素明胶微球的制备及体外释药特性

目的对阿霉素明胶微球(ADM-GMS)的制备工艺及体外释药特性进行研究.方法用乳化交联法制备阿霉素明胶微球,采用正交试验设计法考察影响制备工艺的各因素.用扫描电镜观察微球表面形态.并对阿霉素明胶微球的粒径大小及其分布、载药量、包封率、体外释药、稳定性等进行了测定.结果微球形态圆整,大小均匀、表面光滑,平均粒径为69.154μm,87.32%的微球粒径分布在50～120μm.微球平均载药量为2.13%,包封率为42.62%.微球体外降解和释药特性满足要求,且37℃下性质稳定.结论该制备工艺稳定,制得的阿霉素明胶微球有望成为临床用动脉栓塞术治疗癌症的新制剂.     

海昆肾喜防治阿霉素肾病大鼠作用的实验研究

目的：探讨海昆肾喜（FPS）防治阿霉素肾病大鼠肾小球硬化的作用机制。方法：将54只Wistar雄性大鼠，随机分为6组，每周测体重，每2周收集24小时尿量，测24小时尿蛋白定量，10周后处死，分离血清观察各项生化指标，应用HE、六胺银染色及Mallory染色，观察肾组织病理学改变，实验数据用SPSS11．0进行统计学处理。结果：海昆肾喜能够减轻阿霉素肾病大鼠肾组织病理损害，增加阿霉素肾病大鼠体重，减少尿蛋白排泄、提高血TP和Alb水平，降低BUN和Scr，改善肾功能。结论：海昆肾喜能减轻阿霉紊肾病大鼠的肾脏病理损害、改善肾功能、调节血脂紊乱和升高白蛋白水平，对阿霉素肾病大鼠肾小球硬化有一定的保护作用。     

不同剂量电离辐射对大肠癌细胞株HCT-8的多柔比星敏感性的研究

目的研究不同剂量电离辐射作用后多柔比星（阿霉素）对大肠癌多药耐药细胞株HCT-8细胞毒活性的影响,进一步探讨逆转大肠癌多药耐药性的方法.方法体外培养大肠癌细胞株HCT-8,以400 ng/ml阿霉素作为HCT-8细胞耐药模型刺激浓度制备细胞耐药模型.实验模型分为5组：假照射组;大剂量组（2 Gy）;0.05 Gy＋2 Gy组;0.1 Gy＋2 Gy组;0.2 Gy＋2 Gy组.采用MTT法测定给予阿霉素后大肠癌多药耐药细胞株HCT-8的存活率.结果与假照射组相比,2 Gy大剂量照射组及0.2 Gy＋2 Gy组照射后HCT-8细胞存活率明显降低（P＜0.05）,先给予低剂量照射（0.05 Gy,0.1 Gy）后,再给予大剂量照射,HCT-8细胞存活率降低更明显（P＜0.01）.与单纯2 Gy大剂量照射组比较,0.1 Gy＋2 Gy组HCT-8细胞存活率明显降低（P＜0.05）.结论先给予低剂量照射（0.1 Gy）后,再给予大剂量照射,可提高大肠癌多药耐药细胞株HCT-8对阿霉素的敏感性.     

miz1基因siRNA表达载体的构建及影响HeLa细胞对阿霉素的敏感性

目的:构建针对转录因子miz1基因的siRNA表达载体,观察其影响HeLa细胞对阿霉素的敏感性.方法:设计并合成针对miz1基因的siRNA寡核苷酸,克隆入siRNA表达载体,脂质体瞬时转染入HeLa细胞中,用RT-PCR检测miz1基因的mRNA表达水平,用 Western Blot检测Miz1蛋白表达水平,用MTT法检测HeLa细胞活力.结果:DNA测序和酶切鉴定证明针对miz1基因的siRNA表达载体构建成功,RT-PCR和Western Blot表明其能降低miz1基因的mRNA表达和蛋白表达. 与对照组相比,转染上述干扰载体后的HeLa细胞,在加入阿霉素12 h后,细胞活力明显降低(P＜0.05),并且干扰质粒与阿霉素存在交互作用(P＜0.05),以加入阿霉素12 h和24 h后最明显.结论:针对miz1基因的siRNA能够抑制人宫颈癌细胞系HeLa中miz1基因的表达并能增强HeLa细胞对阿霉素的敏感性.