Recombinant human alpha fetoprotein synergistically potentiates the anti-cancer effects of 1′-S-1′-acetoxychavicol acetate when used as a complex against human tumours harbouring AFP-receptors

Purpose

Previous in vitro and in vivo studies have reported that 1′-S-1′-acetoxychavicol acetate (ACA) isolated from rhizomes of the Malaysian ethno-medicinal plant Alpinia conchigera Griff (Zingiberaceae) induces apoptosis-mediated cell death in tumour cells via dysregulation of the NF-κB pathway. However there were some clinical development drawbacks such as poor in vivo solubility, depreciation of biological activity upon exposure to an aqueous environment and non-specific targeting of tumour cells. In the present study, all the problems above were addressed using the novel drug complex formulation involving recombinant human alpha fetoprotein (rhAFP) and ACA.

Experimental Design

To study the synergistic effect of both agents on human cancer xenografts, athymic nude (Nu/Nu) mice were used and treated with various combination regimes intraperitoneally. Serum levels of tumour markers for carcinoembryonic antigen (CEA) and prostate specific antigen (PSA) were assessed using sandwich ELISA. IHC and Western blotting were also conducted on in vivo tumour biopsies to investigate the involvement of NF-κB regulated genes and inflammatory biomarkers. Quantification and correlation between drug efficacies and AFP-receptors were done using IF-IC and Pearson''s correlation analysis.

Results

Mice exposed to combined treatments displayed higher reductions in tumour volume compared to stand alone agents, consistent with in vitro cytotoxicity assays. Milder signs of systemic toxicity, such as loss in body weight and inflammation of vital organs were also demonstrated compared to stand alone treatments. Tumour marker levels were consistent within all rhAFP/ACA treatment groups where levels of CEA and PSA were initially elevated upon commencement of treatment, and consecutively reduced corresponding to a decrease in tumour bulk volume. Both IHC and Western blotting results indicated that the combined action of rhAFP/ACA was not only able to down-regulate NF-κB activation, but also reduce the expression of NF-κB regulated genes and inflammatory biomarkers. The efficacy of rhAFP/ACA complex was also found to be weakly negatively correlated to the level of surface AFP-receptors between tumour types.

Conclusions

This drug complex formulation shows great therapeutic potential against AFP-receptor positive tumours, and serves as a basis to overcome insoluble and non-specific anti-neoplastic molecules.     

高良姜饮片HPLC指纹图谱及高良姜素含量测定研究

目的:采用高效液相色谱法建立高良姜饮片的HPLC指纹图谱,同时测定样品中高良姜素的含量,为其质量控制提供新方法。方法:以Phenom enex C18柱(4.6 mm×250 mm,5μm)为色谱柱,流动相为乙腈-0.2%磷酸溶剂系统,线性梯度洗脱,检测波长为280 nm(高良姜素含量测定为235 nm);采用指纹图谱相似度软件进行数据分析。结果:确定了高良姜饮片9个共有峰,建立了14批药材的共有图谱,并比较了样品中高良姜素的含量。结论:指纹图谱信息结合有效成分含量测定,可较为全面评价高良姜饮片的质量。     

高良姜油抗实验性胃溃疡作用及其机制研究

[目的]研究高良姜油抗胃溃疡（GU）的作用及其机制。[方法]ICR小鼠随机分成7组,分别为：正常对照组,模型组,奥美拉唑组（0.014 g/kg）,良附丸组（2 g/kg）,高良姜油高、中、低剂量（8、42、ml/kg）组,预防给药6d后,采用利血平致GU小鼠模型,收集小鼠胃液,测定胃液量、总酸度、胃蛋白酶活性及血清一氧化氮（NO）、超氧化物歧化酶（SOD）及丙二醛（MDA）水平。[结果]与模型组相比,高良姜油高、中剂量组,良附丸组,奥美拉唑组均能抑制利血平致GU小鼠模型胃液量、总酸度及胃蛋白酶活性,且能增加血清NO水平和SOD活性,降低MDA水平;但中剂量组胃蛋白酶活性变化无统计学意义。[结论]高良姜油抗GU作用机制可能与其抑酸、抗氧化、增强保护因子有关;高良姜油高剂量抗氧化与增加血清NO水平的作用与良附丸相当。     

高良姜素提取工艺改进研究

目的优化从中药高良姜中提取高良姜素工艺研究.方法采用乙醇提取,石油醚脱脂,氯仿萃取,上聚酰胺柱得到高良姜素.采用紫外、红外方法进行检测.结果运用聚酰胺柱层析一次分离,获得有效成分高良姜素.结论本研究优化了高良姜提取工艺.     

草果古今药方纵横

中药与方剂是中医学理法方药中的重要组成部分,也是中医用于防治疾病的重要手段。数千年来,经过前人长期的实践与认识,从药物的认识,到单方、复方的应用,沉积了无数先人的智慧成果,是中医药学留给世人的一个伟大宝库。文章通过综合古今对于草果的记载,并对其出处、功能、主治范围进行分析,一方面不仅可以反映历代对于草果的应用与发展演进过程,集中体现医家组方之思路以及对于疾病的综合思维,同时通过对草果相对完整而系统的论述,更可进一步作为临床研究的基础。     

益智仁中总黄酮含量测定

目的：考察不同产地益智仁中总黄酮的含量,为益智仁的质量标准研究提供依据.方法：用AlCl3-NaNO2-NaOH显色,利用紫外分光光度法测定益智仁中总黄酮的含量,检测波长为510 nm.结果：在19.2～38.4 μg/ml范围内,吸收度与浓度呈良好的线性关系,回归方程：A=0.015 1c＋0.038 1,r=0.999 8;回收率为100.44%,RSD=0.51%.结论：该方法可靠、结果稳定、重现性好,可作为益智仁中总黄酮的含量测定方法.     

高良姜与香附配伍的不同提取方法的研究

目的:通过测定高良姜与香附配伍的不同提取方法中高良姜素的含量,确定高良姜与香附配伍的提取方法。方法:采用HPLC法,以乙腈-0.5%磷酸(42:58)为流动相,检测波长:266nm,流速:1.0ml·min-1;结果:高良姜与香附的提取方法为乙醇合并提取时,高良姜素的含量高于其他提取方法。结论:高良姜与香附的提取方法为乙醇合并提取时,有助于高良姜中有效成分高良姜素的溶出。     

益智仁中多糖的含量测定

目的 考察不同产地益智仁中多糖的含量,为中药益智仁的质量标准研究提供一定的依据.方法 用苯酚硫酸显色,利用紫外分光光度法,测定益智仁中多糖的含量,检测波长488 nm.结果 在3.45～10.35 mg·L-1范围内,吸收度与浓度呈良好的线性关系,其回归方程:A=0.078C 0.0291,相关系数r=0.999 4;回收率为100.41%,RSD=0.75%.结论 该方法可靠、结果 稳定、重现性好,可作为益智仁中多糖的含量测定方法.     

Evaluation of the antifungal potential of Brazilian Cerrado medicinal plants

Therapeutic limitations, development of fungal drug resistance, drug-related toxicity, drug interactions and insufficient bioavailability of the currently available antifungal drugs have made the development of drugs necessary that would be able to treat the emerging fungal infections. The Cerrado is the second greater biome of Brazil and it was identified as one of the most distinguished biomes of South America, becoming an important source of innovative vegetal molecules to treat several conditions. Thus, the objective of this study was to evaluate the antifungal potential of Cerrado plants, mainly those used to treat infections and wounds. A total of 57 extracts were screened by the agar-well diffusion technique against Candida albican s and Trichophyton rubrum . The most promising extracts were tested in smaller concentrations and their minimal inhibitory concentrations (MIC) were determined by microdilution method. Results were analysed statistically by anova tests. Extracts of Kielmeyera coriacea , Renealmia alpinia , Stryphnodendron adstringens and Tabebuia caraiba were very active against T. rubrum , presented geometric means of the MIC values between 170.39 and 23.23 μg ml⁻¹. Extracts of Cerrado plants are of particular interest as source of new agents for the treatment of dermatophytic infections.     

草果古今药方纵横

中药与方剂是中医学理法方药中的重要组成部分,也是中医用于防治疾病的重要手段。数千年来,经过前人长期的实践与认识,从药物的认识,到单方、复方的应用,沉积了无数先人的智慧成果,是中医药学留给世人的一个伟大宝库。文章通过综合古今对于草果的记载,并对其出处、功能、主治范围进行分析,一方面不仅可以反映历代对于草果的应用与发展演进过程,集中体现医家组方之思路以及对于疾病的综合思维,同时通过对草果相对完整而系统的论述,更可进一步作为临床研究的基础。