长春新碱通过下调SODD蛋白表达诱导白血病细胞凋亡新机制研究

 目的研究死亡结构域沉默子(silencer of death domains,SODD)、caspase3、caspase8及caspase9在长春新碱诱导Jurkat白血病细胞凋亡过程中的变化,探讨长春新碱诱导肿瘤细胞凋亡的新机制。方法采用Annexin V/PI双标记流式细胞术检测长春新碱(VCR)作用后Jurkat细胞凋亡发生率;采用免疫印迹法分析SODD、caspase3、caspase8及caspase9蛋白表达的变化;ELISA酶联免疫吸附技术检测VCR作用Jurkat细胞后TNF-α分泌的变化;采用RT-PCR检测VCR对细胞,TNFR1mRNA表达的调节。结果Jurkat白血病细胞SODD蛋白高表达且高表达的SODD蛋白抑制肿瘤细胞凋亡,VCR能特异下调SODD蛋白的表达,有效诱导Jurkat细胞凋亡,但并不影响细胞TNF-α的分泌及TNFR1的表达;VCR诱导细胞凋亡过程中caspase3、caspase8酶原呈时间依赖性逐渐被水解剪切,而caspase9在该凋亡过程中无明显变化趋势。结论VCR下调SODD蛋白表达并启动外源性凋亡途径caspases级联(caspase8、caspase3),最终诱导Jurkat细胞凋亡,且VCR下调SODD蛋白的表达无需激活TNF/TNFR1信号途径即可导致凋亡的发生。     

化疗药物对荷人胆管癌裸小鼠移植瘤的药效试验

目的：观察化疗药物对荷人胆管癌裸小鼠移植瘤的治疗效果。方法：采用人中分化胆管癌裸小鼠移植瘤模型。将45只荷人胆管癌裸小鼠分为各治疗组和对照组,裸小鼠治疗组每组5只,对照组10只。分组当天称鼠重,由尾静脉用药1次。给药剂量：丝裂霉素(MMC)4mg／kg,阿霉素(ADM)10mg／kg,长春新碱(VCR)0．20mg／kg,泰素(TAXOL)20mg/kg,顺铂(DDP)10mg／kg,环磷酰胺（CTX)120mg／kg,5-氟脲嘧啶(5-Fu)120mg/kg,NS为0．2mL／只。第21天处死裸小鼠,称鼠重,计算体重变化。治疗期间每周测肿瘤瘤径2次,计算相对肿瘤增殖率T／C(％)。结果：MMC,ADM,VCR,TAXOL,DDP和CTX组第21天相对肿瘤增殖率分别为：1％,23％,39％,47％,51％和86％,裸小鼠体重增加分别为：21％,0％,11％,1％,14％和8％。5-Fu组80％裸小鼠死亡,有药物毒性反应。结论：MMC,ADM,VCR,TAXOL,DDP对荷人中分化胆管癌裸小鼠的移植瘤有明显的抗癌作用,CTX无效。     

Treatment of recurrent malignant supratentorial gliomas with the association of procarbazine,thiotepa and vincristine: A phase II study

Summary Twenty patients previously treated with surgery, radiation therapy and chemotherapy with a nitrosourea for malignant supratentorial gliomas received a combination of procarbazine, thiotepa and vincristin (P.T.V.) at tumor recurrence. Procarbazine was given at a dose of 100 mg/m² per os from day 1 (D1) to day 15 (D15), thiotepa was administered IV at a dose of 35–45 mg/m² at D1, and vincristine at a dose of 1,4 mg/m² at D1 and D8. Courses of therapy were repeated every four weeks. Tolerance was evaluated in 20 patients. Three patients developed peripheral neuropathy after 2 or 3 courses of vincristin which was then discontinued. Blood toxicity over grade I occurred in 8 patients (40%). One patient developed a grade IV pancytopenia. All 20 patients could be evaluated for therapeutic response. A partial response was noted in 3 patients (15%): 1 glioblastoma multiforme, 1 anaplastic oligodendroglioma and 1 anaplastic astrocytoma. In these three patients time to tumor progression was 10, 11 + and 5 months, respectively. Stabilization lasting 4 months was observed in one patient (anaplastic astrocytoma). Estimated median duration of survival for the entire group was 4,5 months following the onset of PTV (13 months following the date of histology).     

Concurrent decline of several antioxidants and markers of oxidative stress during combination chemotherapy for small cell lung cancer

Objectives

To investigate the oxidant effects of adriamycin-containing chemotherapy (CT), we evaluated various antioxidants, total antioxidant capacity (TRAP) and different parameters of oxidative and nitrosative stress during combination CT.

Design and methods

Blood samples were obtained from 16 small cell lung cancer patients at baseline and several times during the first, second and sixth CT cycles.

Results

There were significant decreases in serum urate and serum proteins during all cycles, serum TRAP during the first two cycles, plasma ascorbic acid and serum TBARS during the first cycle, and serum conjugated dienes and plasma alphatocopherol during the last cycle. The baseline levels of tocopherols increased significantly between the first and sixth CT cycles. Higher levels of baseline plasma thiols were associated with better overall survival (p  = 0.008).

Conclusions

Adriamycin-containing CT causes significant oxidative stress as implied by reduced levels of protective antioxidants. Long-term CT treatment seems to enhance lipid peroxidation.     

长春新碱固体脂质纳米粒的制备工艺优化

目的制备长春新碱固体脂质纳米粒(VCR-SLN)并优化其处方组成和制备工艺。方法单因素考察筛选载体、稳定剂及制备工艺,用正交试验进行优化,以包封率、载药量和粒径为指标,筛选最佳处方和制备工艺,并对在优化条件下制备的VCR-SLN进行质量评价。结果以单硬酸酯甘油酯为载体,大豆卵磷脂、泊洛沙姆188为乳化剂,采用复乳-溶剂扩散法制备得VCR-SLN,其平均粒径为156.3 nm,包封率为55.12%,载药量为3.09%。结论复乳-溶剂扩散法适用于制备VCR-SLN。     

The brain slice chamber,a novel variation of the Boyden Chamber Assay,allows time-dependent quantification of glioma invasion into mammalian brain <Emphasis Type="Italic">in vitro</Emphasis>

Summary Glioma cell invasion occurs in a complex micromilieu consisting of neural and glial cells, myelinated fiber tracts, blood vessels and extracellular matrix proteins. The present work describes the brain slice chamber (BSC) as a novel experimental model for assessing invasion of glioma cells into adult mammalian white and gray matter on the basis of the well known Boyden chamber system. As a matrix for invasive tumor cells we used freshly prepared brain tissue from adult pigs. The tissue was sectioned into 40 m slices that were mechanically fixed to a millipore filter. The neural structures and the three-dimensional architecture of the slice was preserved as verified by immunohistochemistry, light- and electron microscopy. Human U-373 and U87 astrocytoma cells stably transfected with green fluorescent protein (GFP) were assessed for their invasiveness into the brain-slices during a 24 h period. Invasion of U-87 GFP cells was quantified at different time intervals by confocal laser scanning microscopy showing more intense invasion into white compared to gray matter. Two cytostatics (vincristin and paclitaxel) which both are known to affect the cytoskeleton, inhibited glioma cell invasion in a dose dependent manner, which makes the presented model system suitable for functional experiments. In conclusion, the BSC represents a valid and rapid experimental model that may be used to describe the invasive behavior of glioma cells within the preserved three-dimensional structure of mammalian brain tissue in vitro.