Additive effects of earthworm extract with HpD-laser on the enhancement of reactive oxygen production and the inhibition of[~3H]—thymidine.uptake in mouse ascitic tumor cells

The effects of combined use of earthworm extract(912)and HpD-laser on the produc-tion of reactive oxygen and the biosynthesis of DNA in S_(180) tumor cells were studied throughchemiluminescence measurement and[~3H]-TdR incorporation assay.The results showed that as com-pared with the control,the intensity of chemiluminescence emitted by tumor cells treatedsimultaneously with 912 and HpD-laser was enhanced more than ten-folds,while that treated with912 or HpD-laser alone was increased only 2～4 folds.The[~3H]-TdR incorporation into tumorcells of the former group was inhibited upto 74.1%,and that of the latter groups decreased onlyby 42.2% and 40.0%,respectively.In accordance with these biochemical changes,the ultrastructuraldamage of tumor cells of the former,combinedly treated group appeared to be the most serious.This suggests an additive effect of 912 with HpD-laser on tumor cells.In addition,if free radicalscavengers,such as catalase and superoxide dismutase,were added to the reaction systembefore chemiluminescence assay,the luminescent enhancement effect mentioned above was dramaticallyalleviated,implying the presence of O_2~ and H_2O_2 in the system.Therefore,as to the toxic effecton tumor cells,912 and HpD-laser are not only additive in efficiency,but also similar in theunderlying mechanism of action.     

蚯蚓纤溶酶的纯化及pH值、温度和胰蛋白酶对其纤溶活性的影响

目的纯化并鉴定蚯蚓纤溶酶 ,研究pH、温度和胰蛋白酶对其纤溶活性的影响。方法以赤子爱胜蚓为材料 ,采用硫酸铵盐析、DEAESepharoseCL 6B离子交换色谱、SephadexG 75凝胶过滤和制备电泳等分离技术纯化蚯蚓纤溶酶 ;利用纤维平板和聚丙烯酰胺凝胶电泳研究pH、温度和胰蛋白酶对蚯蚓纤溶酶纤溶活性的影响。结果分离得到纤溶酶 4种单一组分 :EFE 1、EFE 2、EFE 3、EFE 4 ;分子量分别为 2 70 0 0、2 80 0 0、2 30 0 0、330 0 0 ;4种组分对温度的适应范围较广 ;pH值对其纤溶活性的影响不尽相同 ,但都在pH低于 1.5时纤溶活性完全丧失 ;胰蛋白酶对纤溶酶 4种组分均无降解作用 ,对其纤溶活性无影响。结论纤溶酶口服药制剂宜制成肠溶型     

Full-driving soft robotic colonoscope in compliant colon tissue

Robotic colonoscopy is an efficient examination method for finding malignant tumour in its early stage. This research developed a novel robotic endoscope with 13?mm diameter, 105?mm length and 22.3?g weight. A contact biomechanical model is proposed to increase the locomotion safety and efficiency in the soft tissue. The model shows that the friction difference between the robot and the tissue is a key factor to locomotion capability. A soft, full bellow with excellent compatibility was designed to package the robot body. The bellow increased the static friction and decreased the kinetic friction given the change in the contact state. The bellow is divided into three segments. Each segment is composed of a linear locomotor with micromotor, turbine-worm and wire wrapping–sliding mechanism. The robot is tested in in vivo pig colon, which revealed an excellent locomotion capability and safety in soft tissues.     

Earthworm biomarker responses on exposure to commercial cypermethrin

Cypermethrin is a synthetic pyrethroid insecticide used worldwide in agriculture, home pest control, disease vector control, and food safety. It accumulates in soil. Therefore, traces of cypermethrin may frequently appear in vegetables grown in contaminated soil. There is a push now to develop biomarkers as early warning indicators of environmental pollution. In this study, DNA damage (tail DNA%, tail length, and olive tail moment), the micronucleus, neutral red retention (NRR) time, and pinocytic adherence ability of coelomocytes were investigated in Pheretima peguana earthworms exposed to cypermethrin in filter paper tests. The NRR time of earthworm coelomocytes decreased significantly at a concentration of 3.5 × 10^?3 µg · cm^?2 (1/100 LC₅₀) after 48 h exposure, with a highly negative correlation with cypermethrin concentration. Pinocytic adherence ability of coelomocytes also declined significantly at a cypermethrin concentration of 3.5 × 10^?2 µg · cm^?2 (1/10 LC₅₀). The DNA damage to earthworm coelomocytes (tail DNA%, tail length, and olive tail moment) increased considerably at the highest concentration (3.5 × 10^?1 µg · cm^?2) although the correlation between tail DNA% and cypermethrin concentration was low. Thus, physiological biomarkers were more sensitive than the genotoxic effects in earthworms exposed to commercial cypermethrin. Although a suite of earthworm biomarkers could be used to evaluate cypermethrin terrestrial pollution, the NRR test is easier to conduct and a more sensitive indicator. © 2013 Wiley Periodicals, Inc. Environ Toxicol 30: 597–606, 2015.     

Interaction of citrate‐coated silver nanoparticles with earthworm coelomic fluid and related cytotoxicity in Eisenia andrei

Understanding the interaction of nanoparticles with biological fluid is important for predicting the behavior and toxicity of nanoparticles in living systems. The earthworm Eisenia andrei was exposed to citrate‐coated silver nanoparticles (cAgNPs), and the interaction of cAgNPs with earthworm coelomic fluid (ECF), the cytotoxicity of cAgNPs in earthworm coelomocytes was assessed. The neutral red retention assay showed a reduction in lysosomal stability after exposure. The toxicity of silver ions dissolved from cAgNPs in the soil medium was not significant. The aggregation and dissolution of cAgNPs increased in ECF, which contains various electrolytes that alter the properties of nanoparticles, and their subsequent toxicity. Microscopic and dissolution studies demonstrated that the aggregation of cAgNPs rapidly increased, and readily dissolved in ECF. The bioavailability of cAgNPs to earthworms induced lysosomal cytotoxicity. This is the first report to test the interaction and lysosomal cytotoxicity of nanoparticles in earthworm biofluids. Copyright © 2014 John Wiley & Sons, Ltd.     

蚯蚓纤溶酶增加胃癌细胞对放射线敏感性的实验研究

目的探讨蚯蚓纤溶酶对胃癌细胞株MGC803的放射增敏作用及机制。方法采用克隆形成实验观察蚯蚓纤溶酶的放射增敏作用;流式细胞术观察药物对胃癌细胞株MGC803周期分布的影响;RT-PCR法检测药物对细胞内硒谷胱甘肽过氧化物酶(SeGPx)基因表达的影响。结果蚯蚓纤溶酶对胃癌细胞有放射增敏效应,100uku/L、300uku/L蚯蚓纤溶酶的放射增敏比分别为1.06和1.14。射线照射后MGC803细胞发生G1期阻滞,蚯蚓纤溶酶对细胞G1期阻滞无明显影响;PCR检测发现射线可诱导的SeGPxmRNA表达下调,蚯蚓纤溶酶能够增强射线诱导的SeGPxmRNA表达下调作用。结论蚯蚓纤溶酶能够增加胃癌细胞对放射线的敏感性,增敏机理可能与药物下调细胞内抗氧化酶SeGPx基因表达有关。     

蚯蚓提取物中蛋白质含量测定方法的研究

目的寻找能够精确反映蚯蚓提取物中蛋白质含量的测定方法。方法采用改良Lowry法和考马斯亮蓝染料结合法(Bradford法)测定蚯蚓提取物蛋白质含量;分别使用2法测定标准牛血清白蛋白(BSA)水溶液在碱性蛋白酶水解前后的蛋白质含量。结果改良Lowry法的测定结果明显高于Bradford法。BSA水溶液酶解前,2法测定的蛋白质含量相当;使用改良Lowry法,BSA水溶液酶解前后的蛋白测定值相近;而使用Bradford法测定BSA水溶液蛋白质含量时,酶解后的测定值较酶解前出现大幅下降。结论在测定多肽含量多的供试品中蛋白质含量时,改良Lowry法测定更为精确。蚯蚓提取物中存在大量的多肽成分,所以对其进行蛋白质含量测定时,改良Lowry法能够更加精确、真实地反映蚯蚓提取物的蛋白质含量。     

补肾活血方对恒河猴PCO模型卵巢COX-1mRNA表达的影响

目的探讨补肾活血方对恒河猴多囊卵巢(PCO)模型卵巢局部环氧化酶-1(COX-1mRNA)表达的影响。方法选择健康育龄雌性恒河猴(6只)随机分为正常对照组、模型对照组、补肾活血方组;采用皮下注射丙酸睾丸酮(T)和绒毛膜促性腺激素(HCG)建立PCO模型,分别灌胃给予生理盐水、补肾活血方,采用原位杂交技术观察各组卵巢局部COX-1mRNA表达的变化。结果各组卵巢组织均可见COX-1基因表达的信号,以颗粒细胞表达最丰富;其中模型组卵泡膜细胞呈现异常高表达;补肾活血方组表达水平接近正常对照组。结论恒河猴PCO模型卵巢局部COX-1mRNA异常表达,可能与多囊卵巢综合征(PCOS)发病有关;补肾活血方有可能通过降低COX-1mRNA的表达而达到改善PCOS无排卵的状况。     

Ecotoxicity of chromium (III) to Eisenia fetida, Enchytraeus albidus, and Folsomia candida

Despite growing concerns about the potential adverse effects of elevated chromium concentrations in the environment, only a few terrestrial toxicity data are available. Therefore, chronic toxicity of chromium(III) was assessed for Eisenia fetida, Enchytraeus albidus, and Folsomia candida, the three invertebrates for which standard test protocols are available. The 21-day EC(50) for the cocoon production of E. fetida was 892 (679-1110) mg Cr/kg dry weight. Based on the reproduction, a 42-days EC(50) of 637 (355-791) mg Cr/kg dry weight was observed for E. albidus, while for F. candida a 28-day EC(50) of 604 (254-3380) mg Cr/kg dry weight was noted. Although the presented data can be considered a step forward in the assessment of the potential risks of chromium(III) in terrestrial environments, further research is needed to evaluate the influence of soil parameters on the toxicity of chromium and to quantify the effect of aging on bioavailability.     

水蛭地龙萃取液对大鼠局灶性脑缺血再灌注损伤粘结合蛋白多糖-1的影响

目的：探讨水蛭地龙萃取液对急性脑梗死粘结合蛋白多糖-1的表达及可能的抗炎机理。方法：采用线栓法制作大鼠大脑中动脉（MCAO）阻断模型，用HE染色、免疫组织化学方法，观察水蛭地龙萃取液对脑缺血2h再灌注4、24、72h和7d的粘结合蛋白多糖-1阳性细胞的分布、数量及缺血脑组织的病理变化。结果：水蛭地龙萃取液组各时间点均可上调粘结合蛋白多糖-1的表达（P〈0．01），其中72h最明显；水蛭地龙萃取液治疗24、72h时间点较对照组多核炎性细胞的浸润明显减少，而4h点两组炎性细胞浸润均不明显，7d水蛭地龙萃取液组能明显减少单核细胞的浸润。结论：水蛭地龙萃取液可以上调粘结合蛋白多糖-1的表达，粘结合蛋白多糖-1的上调能减少炎性细胞的浸润。