大肠癌细胞的药敏试验结果分析及临床意义

目的通过测定大肠癌患者对5-氟脲嘧啶（5-Fu）、顺铂（DDP）、阿霉素（ADM）、丝裂霉素（MMC）、奥沙利伯（OXA）及联用化疗药物的体外敏感性试验，探讨肿瘤药敏试验对大肠癌个体化疗的应用价值。方法采用组织块培养-终点染色-计算机图像分析法（TECIA）。结果36例大肠癌细胞对试验化疗药物的敏感性由高到低依次为OXA、F＋M（5-Fu＋MMC）、F＋D（5-Fu＋DDP）及DDP、5-Fu、O＋F（5-Fu＋OXA）、MMC、ADM。结论TECIA法的体外肿瘤药敏试验在大肠癌的l临床用药及个体化化疗方面具有重要的指导意义。     

通络救脑口服液对实验性类AD大鼠血清细胞因子含量的影响

目的探讨大鼠大脑海马立体定向注射β-淀粉样蛋白1-40对血清TNF-α和IL-1β含量的影响及通络救脑口服液对其干预作用。方法140只Sprague-Dawley(SD)大鼠随机分为空白对照组、假手术组、阴性对照组、阳性对照组、实验组(12,24,48 mg.kg-1.d-1)共7组。采用大脑海马立体定向注射β-淀粉样蛋白1-40诱导Alzheimer's病动物模型后,药物干预4周,第5周应用双抗夹心酶联免疫吸附测定检测TNF-α和IL-1β含量。结果模型组TNF-α,IL-1β含量分别为(120.97±7.32)和(88.54±4.07),TNF-α和IL-1β含量显著多于假手术组(TNF-α为85.43±6.55,IL-1β为68.29±4.37,P<0.01),表明模型组TNF-α和IL-1β含量升高;与模型组比较,盐酸多萘哌齐组(TNF-α为106.86±7.09;IL-1β为1.80±0.23)、通络救脑口服液12,24,48 mg.kg-1.d-1剂量组(TNF-α分别为100.08±9.29、99.97±6.09和85.90±4.40;IL-1β分别为(77.56±6.97),(77.85±3.81)和(67.87±2.38)TNF-α和IL-1β含量升高显著减少。结论大鼠海马注射β-淀粉样蛋白1~40后大鼠血清TNF-α和IL-1β含量明显增高,通络救脑口服液可以降低血清TNF-α和IL-1β的含量,从而发挥其抗老年性痴呆的作用。     

The dietary antioxidants resveratrol and quercetin protect cells from exogenous pro-oxidative damage.

In the colorectal epithelium oxidative stress is observed endogenously in premalignant adenoma cells or induced by nutritional factors like fatty acid hydroperoxides (LOOH). Bioactive phenols like resveratrol and quercetin can quench reactive oxygen species and protect from pro-oxidative damage. Our study used colorectal adenoma and carcinoma cell lines to assess antioxidant protective effects of resveratrol and quercetin. It demonstrated that both compounds efficiently protect from oxidative stress induced by LOOH. Effective concentrations (10 microM resveratrol and 1 microM quercetin) can easily be reached in the intestinal lumen after consumption of plant foods or food supplements. Both compounds prevent LOOH-induced formation of intracellular H2O2, stimulation of cyclooxygenase-2 and vascular endothelial growth factor. For reduction of endogenous H2O2 levels in colorectal tumor cells higher antioxidant-concentrations are needed in all cell lines. Quercetin (10 microM) alone even increased H2O2 in LT97 adenoma cells and stimulated VEGF production. Resveratrol and quercetin also induced 10-30% and 40-60% cell loss respectively by apoptosis. In summary, this indicates that resveratrol and quercetin have little protective capacity in absence of exogenous stress. They are however highly efficient in protecting against nutrition induced oxidative stress damage suggesting that this constitutes the major part of their tumor protective activity.     

Protective effect of basil (Ocimum basilicum L.) against oxidative DNA damage and mutagenesis.

Mutagenic and antimutagenic properties of essential oil (EO) of basil and its major constituent Linalool, reported to possess antioxidative properties, were examined in microbial tests. In Salmonella/microsome and Escherichia. coli WP2 reversion assays both derivatives (0.25–2.0 μl/plate) showed no mutagenic effect. Salmonella. typhimurium TA98, TA100 and TA102 strains displayed similar sensitivity to both basil derivatives as non-permeable E. coli WP2 strains IC185 and IC202 oxyR. Moreover, the toxicity of basil derivatives to WP2 strains did not depend on OxyR function. The reduction of t-BOOH-induced mutagenesis by EO and Linalool (30–60%) was obtained in repair proficient strains of the E. coli K12 assay (Nikoli?, B., Stanojevi?, J., Miti?, D., Vukovi?-Ga?i?, B., Kne?evi?-Vuk?evi?, J., Simi?, D., 2004. Comparative study of the antimutagenic potential of vitamin E in different E. coli strains. Mutat. Res. 564, 31–38), as well as in E. coli WP2 IC202 strain. EO and Linalool reduced spontaneous mutagenesis in mismatch repair deficient E. coli K12 strains (27–44%). In all tests, antimutagenic effect of basil derivatives was comparable with that obtained with model antioxidant vitamin E. Linalool and vitamin E induced DNA strand breaks in Comet assay on S. cerevisiae 3A cells, but at non-genotoxic concentrations (0.075 and 0.025 μg/ml, respectively) they reduced the number of H₂O₂-induced comets (45–70% Linalool and 80–93% vitamin E). Obtained results indicate that antigenotoxic potential of basil derivatives could be attributed to their antioxidative properties.     

HPLC法测定双参龙胶囊中人参皂苷Rb1的含量

目的:建立双参龙胶囊中人参皂苷 Rb_1含量测定的高效液相色谱法。方法:采用 Kromasil C_(18)柱(KR100-5C_(18),4.6 mm×250 mm,5μm),乙腈-水(28.7:71.3)为流动相;流速为1.0 mL·min~(-1);检测波长为203 nm。结果:人参皂苷 Rb_1在0.848～4.24μg范围内,线性关系良好(r=0.9991),方法平均回收率为99.3%,RSD=1.2%(n=6)。结论:方法简便,结果准确,可用于双参龙胶囊中人参皂苷 Rb_1的定量。     

高效液相色谱法测定复方吲哚美辛乳膏中吲哚美辛的含量

目的:建立复方吲哚美辛软膏中吲哚美辛含量的高效液相色谱方法。方法:采用 XTerra~C_(18)色谱柱(4.6 mm×150mm,5μm),以 pH 5.0的磷酸盐缓冲液-乙腈(60:40)为流动相,流速为1.0 mL·min~(-1),柱温为30℃,检测波长为320 nm,进样量20μL,用外标法测定。结果:吲哚美辛在6.0～60μg·mL~(-1)范围内线性关系良好(r=0.9999),平均回收率为99.2%,RSD 为0.24%。结论:本方法灵敏度高,操作简便可靠。     

2型糖尿病患者T细胞亚群、TNF-α及IL-10的相关性研究

目的探讨2型糖尿病患者外周血T细胞亚群、血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-10之间的相关性.方法用流式细胞仪分析46例初诊2型糖尿病(DM组)患者和39例正常对照者(NC组)的外周血T细胞亚群水平,双抗夹心酶联免疫吸附法检测血清TNF-α、IL-10的水平.同时检测空腹血糖(FPG)、空腹胰岛素(FIns)、糖化血红蛋白(GHbA1c).结果DM组CD4 、CD4 /CD8份别为(48.89±5.50)%,2.64±0.57明显高于NC组的(34.52±3.04)%、1.52±0.27,差异均有统计学意义(P＜0.05);而CD8 则明显低于NC组[(20.16±1.65)%vs(28.28±3.27)%,P＜0.05];DM组和NC组的血清TNF-o水平[(12.71±5.37)ng/L vs(8.17±2.80)ng/L],差异有统计学意义(P＜0.05);而2组血清IL-10的水平差异无统计学意义[(4.61±1.20)ng/L vs(4.36±0.84)ng/L,P＞0.05].结论机体免疫功能异常与促炎因子的启动在2型糖尿病的发生、发展中发挥着一定的作用.     

HPLC测定丹皮酚软膏（丹皮酚霜）中丹皮酚的含量

目的:采用 HPLC 法测定丹皮酚软膏(霜)中丹皮酚的含量。方法:色谱柱为 Hypersil C_(18)柱(150 mm×4.6 mm,5 μm);流动相:甲醇-水(50:50);流速1.0 mL·min~(-1);检测波长:274 nm。结果:丹皮酚进样量在0.05～0.54 μg范围内呈良好线性关系(r=0.9998),平均回收率(n=6)为98.7%,RSD 为2.0%。结论:本法准确,重现性好。     

抗副溶血弧菌TLH蛋白多克隆抗体的制备及其ELISA双抗体夹心检测法的研究

目的分别制备抗副溶血弧菌及其不耐热溶血毒素(thermolabile hemolysin,TLH)的多克隆抗体,建立检测副溶血弧菌TLH的酶联免疫吸附试验(ELISA)双抗体夹心法。方法以副溶血弧菌及其TLH分别免疫健康雄性新西兰大白兔和雌性BALB/C小鼠,制备兔抗副溶血弧菌多克隆抗体及小鼠抗TLH多克隆抗体,以间接ELISA法检测两种抗体效价,并以棋盘法筛选两种多克隆抗体用于夹心检测TLH的效价。结果兔抗副溶血弧菌多克隆抗体效价达1∶6400,鼠抗TLH多克隆抗体效价为1∶102 400,经筛选确定以1∶800稀释的兔抗副溶血弧菌多克隆抗体包被酶标板,与1∶1600稀释的鼠抗TLH抗体建立了ELISA双抗体夹心法。结论兔抗副溶血弧菌多克隆抗体可与TLH呈特异性反应,以此建立的双抗体夹心法检测副溶血弧菌TLH可获较满意结果,此实验可为进一步建立简便、快速的副溶血弧菌检测方法奠定基础。     

高效液相色谱法测定葫芦素B人血清白蛋白纳米粒中药物含量

目的:建立葫芦素B人血清白蛋白纳米粒中药物含量的测定方法。方法:采用Diamonsil C18柱(200mm×4.6 mm,5μm);以乙腈-水-磷酸(45∶55∶0.1)作为流动相;流速为1 mL.min-1;检测波长为228 nm;柱温30℃。结果:在本实验条件下蛋白纳米粒中辅料对葫芦素B的测定无干扰,在1.0~20 mg.L-1浓度范围内线性关系良好(r=0.999 9,n=5),葫芦素B平均回收率为100.2%,RSD为0.8%(n=3)。结论:本法准确可靠,简便易行,可用于葫芦素B人血清白蛋白纳米粒中药物含量的测定。