Molecular characterization of Echinococcus granulosus in south-eastern Romania: evidence of G1–G3 and G6–G10 complexes in humans

Echinococcus granulosus is the aetiological agent of cystic echinococcosis (CE), which is a public health problem in many eastern European countries, particularly in Romania, where the infection causes a high number of human and animal cases. To shed light on the transmission patterns of the parasite, we performed a genotyping analysis on 60 cyst samples obtained from patients who live in south-eastern Romania and who underwent surgery for liver or lung CE. DNA was extracted from the endocysts or the cyst fluids, and fragments of cytochrome c oxidase subunit 1 and NADH dehydrogenase subunit 1 mitochondrial genes (cox1 and nd1, respectively) were amplified by PCR and sequenced. We found that most of the samples analysed (59/60) belonged to the G1–G3 complex (E. granulosus sensu stricto), which contains the most widespread and infective strains of the parasite. We also identified the first human patient infected by a non-G1–G3 genotype of E. granulosus in this country. As the DNA sequence of this cyst sample showed maximum homology with the G6–G10 complex (Echinococcus canadensis), this is, in all likelihood, a G7 genotype, which is often found in pigs and dogs in most countries of eastern and south-eastern Europe.     

Assessing the risk of epidemic dropsy from black salve use

Epidemic dropsy is a potentially life‐threatening condition resulting from the ingestion of argemone oil derived from the seeds of Argemone mexicana Linn. Exposure to argemone oil is usually inadvertent, arising from mustard cooking oil adulteration. Sanguinarine, an alkaloid present in argemone oil, has been postulated as a causative agent with the severity of epidemic dropsy correlating with plasma sanguinarine levels. Cases of epidemic dropsy have also been reported following the topical application of argemone containing massage oil. Black salve, a topical skin cancer therapy also contains sanguinarine, but at significantly higher concentrations than that reported for contaminated massage oil. Although not reported to date, a theoretical risk therefore exists of black salve inducing epidemic dropsy. This literature review explores the presentation and pathophysiology of epidemic dropsy and assesses the risk of it being induced by black salve.     

加拿大棘球绦虫的基因分型与分子流行病学研究进展

通过比较加拿大棘球绦虫不同基因型的线粒体基因组,尤其是其中的cox1和nad1基因核苷酸序列的差异性,了解加拿大棘球绦虫各基因型的分子遗传标记特征与变异情况,阐明加拿大棘球绦虫基因型在棘球属中的分类地位、命名和进化关系。另外,本文通过对加拿大棘球绦虫各基因型的分子流行病学特征、研究意义、未来研究方向等进行综述,为从事该领域研究的学者和临床工作者提供丰富的分子流行病学信息或资料,并为棘球蚴病分子流行病学调查、预警预报和综合防治策略的制定等提供理论依据和指导。     

Study on the association of helicobacter species with viral hepatitis-induced hepatocellular carcinoma

Helicobacter species are important pathogens and previous studies in mice suggested a link between colonization by Helicobacter hepaticus (H. hepaticus) and hepatocellular carcinoma (HCC). This study aimed at corroborating this potential link in human patients. We used a sensitive and specific Helicobacter ssp PCR assay to screen stool samples from a collective of patients with viral-induced HCC (hepatitis B or hepatitis C) and a control group for presence of Helicobacter ssp DNA. Although retrieving DNA of H. pylori and H. canadensis from stool samples of non-HCC patients, we found no evidence indicating the presence of H. hepaticus in HCC-patients with chronic hepatitis B or hepatitis C. Interestingly we found H. canadensis in a stool sample of a patient presenting with diarrhea. Taken together, our data argue against a pathogenic role of H. hepaticus in viral-induced HCC. Yet, our results do not exclude a role of H. hepaticus in those HCC cases caused by other carcinogens, such as aflatoxin. Moreover, we speculate that H. canadensis might be a novel gastrointestinal pathogen.     

Seasonal Changes in the Concentration of Major Taxanes in the Biomass of Wild Canada Yew (Taxus canadensis. Marsh.)

Abstract

Seasonal variations in taxane concentrations in Taxus canadensis. Marsh. (Canada yew) were measured at 15 sampling dates from September 2004 to September 2005 by clipping 3-year-old shoots from the same 15 plants. Four age classes of needles and stems were separated and individually weighed and analyzed for the three major taxanes in T. canadensis.: 10-deacetylbaccatin III (10-DAB III), 9-dihydro-13-acetylbaccatin III (9-DHAB III), and paclitaxel (PAC). The dry weight fraction of each foliage age class decreased with increasing age; that is, proportionately more young than old foliage (current > current + 1 year > current + 2 year) was present in a 3-year-old stem. The ratio of needle:stem dry weight also decreased with increasing foliage age. Taxane concentrations varied widely, depending on the tissue, its age, and time of year. Peak concentrations were observed for a short period at the end of the growing season (August–September). The lowest taxane levels were found during the period of active growth: April through July. PAC and 9-DHAB III concentrations were higher in needles, but young stems contained the highest amounts of 10-DAB III. Levels of all three taxanes were higher in young than in old stems, likely due to increasing amounts of taxane-poor stem wood. Composite taxane concentrations were calculated for 3-year-old foliage from the component needle and stem weights and taxane analyses. It is suggested that summer harvesting between April and July (the low point in annual taxane levels) yields the poorest quality biomass, and harvesting from August onward is preferable.     

小飞蓬活性成分对乳腺癌MCF-7细胞的作用

目的通过体外实验研究小飞蓬（Erigeron canadensis L.）活性成分对乳腺癌MCF-7细胞生长、凋亡和侵袭的影响。方法以5-氟尿嘧啶（5-FU）作用组为阳性对照组,以不加药组为阴性对照组,小飞蓬组为实验组作用MCF-7细胞。采用CCK8试剂盒测定药物对MCF-7的生长抑制率。绘制在不同药物浓度作用下的细胞生长曲线。观察不同药物浓度作用下细胞的形态学变化。利用免疫细胞化学染色技术检测药物作用后MCF-7细胞Caspase-3、MMP-9表达情况。结果 （1）小飞蓬活性成分可抑制MCF-7细胞的生长,IC50为0.29 mg.mL-1;（2）生长曲线显示,药物作用后的MCF-7细胞生长较对照组缓慢,形态学观察显示细胞逐渐变为长,黏附性减弱;（3）药物作用后的MCF-7细胞Caspase-3的表达强于不加药物组（P〈0.001）,MMP-9的表达明显弱于对照组（P〈0.001）。结论小飞蓬活性物质可抑制MCF-7细胞生长、侵袭,诱导凋亡。     

Single-Laboratory Validation of a Method for the Detection and/or Quantification of Select Alkaloids in Goldenseal Supplements and Raw Materials by Reversed-Phase High-Performance Liquid Chromatography

Abstract

Quality of botanical products and raw materials is important to manufacturers, regulators, researchers, and consumers. Many modern botanical quality-assurance schemes set specifications for select phytochemicals and measure against those specifications as one determinant of quality. While numerous publications describe procedures for determining compounds of interest in plant species, few methods have been systematically evaluated for accuracy, precision, or reliability, and often the analysis of finished products is not within the scope of the method. Hydrastis canadensis. L., commonly referred to as Goldenseal, is an economically important North American medicinal plant that has been subject to adulteration in commerce. The phytochemicals of interest in the plant are the alkaloids hydrastine, berberine, and canadine. Of interest is also palmatine, an alkaloid found in potential adulterant species but not in goldenseal. In this study, goldenseal materials in raw, capsule, and tablet form, including an Echinacea/ Goldenseal combination product, were extracted with acidified water and acetonitrile and their hydrastine, berberine, canadine, and palmitine content determined by HPLC. The analytical method was optimized and evaluated in a single-laboratory validation study. Calibration curves for hydrastine and berberine were linear from 10 to 150 μ g/mL. The limits of detection for palmatine and canadine were determined to be 0.5 μ g/mL, which translates to detection of levels of 0.004% w/w in test samples. Chromatographic resolution was achieved for all analytes in an isocratic 12.5-min chromatographic run employing a binary mobile phase. Triplicate determinations performed on 10 test materials by two analysts on 3 days resulted in relative standard deviations ranging from 0.9% to 3.4%.     

小飞蓬化学成分研究

目的:研究小飞蓬的化学成分。方法:采用HP20大孔树脂、硅胶、Sephadex LH-20和十八烷基硅烷键合硅胶(ODS)等多种色谱技术,对小飞蓬进行分离纯化;根据理化常数和波谱数据鉴定所分离化合物的结构。结果:从小飞蓬醇提物的水部位分离了6个化合物,分别鉴定为:Eugenyl-βPsd(1)、野黄芩苷(2)、木犀草素-7-O--βD-葡萄糖醛酸苷(3)、槲皮素(4)、异槲皮素苷(5)和木犀草素(6)。结论:化合物1、3、5和6为首次从小飞蓬中分离得到。     

Accelerated onset of chronic wasting disease in elk (Cervus canadensis) vaccinated with a PrPSc-specific vaccine and housed in a prion contaminated environment

Chronic wasting disease (CWD) is a fatal prion disease affecting multiple cervid species. Effective management tools for this disease, particularly in free-ranging populations, are currently limited. We evaluated a novel CWD vaccine in elk (Cervus canadensis) naturally exposed to CWD through a prion-contaminated environment. The vaccine targets a YYR disease-specific epitope to induce antibody responses specific to the misfolded (PrP^Sc) conformation. Female elk calves (n = 41) were captured from western Wyoming and transported to the Thorne-Williams Wildlife Research Center where CWD has been documented since 1979. Elk were held in contaminated pens for 14 to 20 days before being alternately assigned to either a vaccine (n = 21) or control group (n = 20). Vaccinated animals initially received two vaccinations approximately 42 days apart and annual vaccinations thereafter. Vaccination induced elevated YYR-specific antibody titers in all animals. Elk were genotyped for the prion protein gene at codon 132, monitored for clinical signs of CWD through daily observation, for disease status through periodic biopsy of rrectoanal mucosa-associated lympoid tissue (RAMALT), and monitored for YYR-specific serum antibody titres. Mean survival of vaccinated elk with the 132MM genotype (n = 15) was significantly shorter (800 days) than unvaccinated elk (n = 13) of the same genotype (1062 days; p = 0.003). Mean days until positive RAMALT biopsy for 132MM vaccinated elk (6 7 8) were significantly shorter than unvaccinated 132MM elk (990; p = 0.012). There was, however, no significant difference in survival between vaccinated (n = 4) and control (n = 5) elk with the 132ML genotype (p = 0.35) or in timing of positive RAMALT biopsies of 132ML elk (p = 0.66). There was no strong (p = 0.17) correlation between YYR-specific antibody titers and survival time. Determining the mechanism by which this vaccine accelerates onset of CWD will be important to direct further CWD vaccine research.     

加杨芽鳞提取物的抗肿瘤作用研究

[目的]建立杨树芽鳞提取物(PBE)的制备方法,研究其抗肿瘤活性并探讨其作用机制.[方法]采集加杨芽鳞,提取醇溶性成分PBE.以顺铂为阳性对照,通过酸性磷酸酶法检测PBE对体外培养肿瘤细胞的抑制作用,并进行腹腔接种腹水型肝癌H22细胞的荷瘤小鼠抗癌实验.在光镜与电镜下观察癌细胞的形态学改变.以流式细胞仪和荧光显微镜分析细胞凋亡状况.[结果]PBE对肿瘤细胞系BEL-7402、CNE、NCI-H460、S180的增殖均有明显抑制作用,IC 50分别为8、6、4和12μg/m1,最小有效剂量分别为0.4、0.5、0.8和1.0μg/ml.与顺铂不同的是,PBE对人正常肝细胞L02未见明显抑制作用.体内实验显示PBE可延长荷瘤小鼠的生存期.在PBE作用下癌细胞出现明显形态学变化,包括分裂相细胞减少、细胞表面微绒毛样突起明显增多、细胞连接消失、胞质内积累大量囊泡等.细胞凋亡率未发生明显变化.[结论]PBE对多种肿瘤细胞有显著抑制和杀伤作用.其作用机理可能与改变膜限区室结构、影响细胞表面分子表达、影响微丝系统有关,而非与凋亡途径活化有关.