Chemotype and genotype combined analysis applied to tomato (Lycopersicon esculentum Mill.) analytical traceability

A large number of fresh fruits and vegetables are primary sources of antioxidants; tomato (Lycopersicon esculentum Mill.) is accepted worldwide as a significant source of antioxidant functional compounds (vitamin C, lycopene, rutin). Many cultivars and hybrids of tomato, having different chemical and nutritional characteristics, are available on the market. Tomato cultivars for industrial processing are very different, not only in fruit characteristics (size, shape), but also in lycopene and antioxidant contents. The aim of this study was the chemotyping and genotyping of the tomato varieties Heinz 3402, Leader and Perfectpeel, (1) to evaluate the genetic traceability of these varieties, and (2) to determine whether their functional antioxidants compounds are useful markers of traceability. Principal component analysis (PCA) was first applied to the Random Polymorphic DNA (RAPD) fingerprints, confirming that this approach is a powerful identification method at intra-specific level. Heinz 3402 showed the highest antioxidant activity, followed by Perfectpeel and Leader varieties. Perfectpeel showed the lower lycopene content, while Leader and Heinz 3402 showed significantly higher values (13.68 and 15.78 mg/100 g, fresh weight, respectively). The highest rutin content was observed in Heinz 3402 (12.46 ± 0.69, mg/100 g, fresh weight), followed by Leader (7.87 ± 0.72) and Perfectpeel (2.70 ± 0.68). Antioxidant capacity was significantly correlated with the lycopene and rutin content. Finally, PCA was applied to chemotype data-sets, confirming both mineral content and functional antioxidant compounds as useful markers to unambiguously identify these high-lycopene content varieties.     

New analysis of the toxic compounds from the Androctonus mauretanicus mauretanicus scorpion venom

Scorpion venoms are very complex mixtures of molecules, most of which are peptides displaying different kinds of biological activity. Indeed, these peptides specifically bind to a variety of pharmacological targets, in particular ionic channels located in prey tissues, resulting in neurotoxic effects. Toxins modulating Na+, K+, Ca2+ and Cl(-) currents have been described in scorpion venoms. In this work, we have used several specific antibodies raised against the most lethal scorpion toxins already described to screen the Moroccan scorpion Androctonus mauretanicus mauretanicus venom in order to characterize new compounds. This immunological screening was also implemented by toxicity tests in mice and with mass spectrometry study, providing new informations on the molecular composition of this venom. In fine, we were able to determine the molecular masses of 70-80 different compounds. According to the immunological data obtained, many toxins cross-react with three sera raised against the most lethal alpha-toxins found in North African scorpion venoms, but not at all with those raised against the main beta-toxins from South and North American venoms. Some of the previously described toxins from Androctonus mauretanicus mauretanicus venom could thus be detected by combining immunological tests, toxicity in mice and molecular masses. Among these toxins, one of them, which showed a mild cross-reaction with the serum raised against AaH I (a highly potent toxin from the venom of Androctonus australis), was identified as Amm III and fully sequenced.     

贵州天麻野生与栽培品种的简单序列重复标记鉴定

目的:利用简单序列重复(SSR)分子标记鉴定贵州天麻4个居群的野生和栽培品种.方法:合成8对SSR引物用于天麻基因组DNA的PCR扩增.结果:除引物对16外均能很好地区分野生天麻、无性繁殖天麻及有性繁殖天麻,其中有性繁殖天麻均扩增出2条带,为杂合子;野生天麻及无性繁殖天麻只扩增出1条带,为纯合子.结论:天麻的自交及杂交是形成纯合与杂合的原因.SSR分子标记可有效地区分纯合子及杂合子,从而使其能够在分子水平鉴定野生和栽培天麻.     

半夏不同种质资源AFLP指纹系谱分析及其应用

目的:获得更多半夏DNA指纹信息,验证AFLP与其他方法是否一致,为半夏及其他中药材道地性研究、种质资源鉴定、亲缘关系研究、品种选育和引种栽培提供分子水平生物学依据。方法:选择全国10个主要产地的野生或栽培半夏为材料,采用扩增酶切片段多态性AFLP方法研究,用NTSYSpc 2.1软件包利用UPGMA法进行聚类分析,构建遗传系统树。结果:筛选12对引物,共扩增出1 673条带。进行聚类分析,结果与利用总生物碱含量差异结果相吻合。结论:本研究所获得的特征带和缺失带体现了AFLP的高特异分辨率在中药半夏品种鉴定上的潜力,绘制的系统进化树不同种源之间遗传关系的远近与其总生物碱含量差异趋势一致。     

建立胃癌高发区胃癌血清蛋白指纹图谱诊断模型的探讨

目的 应用表面加强激光解吸离子化飞行时间质谱(SELDI-TOF-MS)技术分析胃癌血清蛋白质指纹图谱,筛选胃癌特异性血清蛋白组合,建立胃癌诊断模型.方法 利用强阴离子交换芯片(Q10)分别对山东省临朐县胃癌高发区36例胃癌患者和46例浅表性胃炎患者的血清蛋白指纹图谱进行检测,采用ZUCIPDAS蛋白芯片数据分析系统,建立高发区胃癌血清蛋白指纹图谱诊断模型,并利用盲筛法验证模型的正确性.结果 通过对26例胃癌患者及37例浅表性胃炎患者血清蛋白指纹图谱进行检测,筛选出差异具有统计学意义的6个蛋白质峰,质荷比分别为8587、6945、8243、3899、7035和9943,并建立胃癌蛋白指纹图谱诊断模型,该模型诊断胃癌的灵敏度达88.5%,特异度达97.3%.在完成建模后,选择19例患者血清进行盲筛,其中胃癌患者10例,浅表性胃炎患者9例.通过盲筛验证,该模型诊断胃癌的灵敏度为80.0%,特异度为88.9%.结论 初步建立了胃癌高发区胃癌血清蛋白指纹图谱诊断模型,该模型诊断胃癌的灵敏度和特异度均达到80%以上,该方法为胃癌的诊断提供了新的思路.     

同种不同产地牛蒡子DNA指纹图谱特征研究

目的 探讨同种不同产地牛蒡子DNA指纹图谱特征。方法 采用RAPD技术对全国四大商品区的中药牛蒡子生品和其中两个商品区的制品进行了DNA指纹图谱特征研究。结果 四大商品区的生品牛蒡子DNA指纹图谱一致,制品牛蒡子的DNA指纹图谱与生品的有较大差异。结论 DNA指纹图谱用于鉴定同种药材的生品结果可靠。     

兔荧光扩增片段长度多态性分析体系的建立

目的:建立兔荧光扩增片段长度多态性(fluorescent amplification fragment length polymorphism,fAFLP)分析方法并对其在实验动物科学中应用进行探讨。方法:本实验以5个品种(系)兔为实验材料,对DNA提取、预扩增和选择性扩增等步骤进行优化并对结果进行检测。结果:5个品种(系)兔的扩增片段数均在120条以上,各品种(系)特异性条带均能找出,5个品种(系)兔能据此进行区分。结论:fAFLP是一种以PCR为基础的DNA指纹技术,为兔的指纹分析提供了一种快速高效的方法,可用于兔的遗传质量检测。     

肠杆菌基因间重复序列PCR指纹图谱技术分析结核分枝杆菌流行情况

目的利用肠杆菌基因间重复序列为引物的聚合酶链反应（enterbacterial repetitive intergenic consensus-PCR，ERIC-PCR）分型技术分析我市结核分枝杆菌流行情况。方法收集2003年9月至2006年5月门诊痰菌阳性标本，培养并提取DNA做ERIC—PCR指纹图，并利用RAPDPHYLIP及Treeview软件对其进行聚类分析。结果122株临床分离株产生42种不同的指纹图，聚类分析结果显示可分为3簇，成簇率76、2％。指纹图谱与患者年龄、菌株的耐药性及耐药种类有关。结论ERIC—PCR具有不需已知核酸序列，分辨效果好，简便快捷等优点，是进行分子流行病学调查的有效工具。我市结核病呈高水平传播，主要为耐药菌株并以中青年患者传播为主。     

Analysis of B cell subsets following pancreatic islet cell transplantation in a patient with type 1 diabetes by cytometric fingerprinting

Manual gating of bivariate plots remains the most frequently used data analysis method in flow cytometry. However, gating is operator-dependent and cumbersome, particularly with the increasing complexity of modern multicolor immunophenotyping data. A method that can remove operator bias, enable systematic and thorough analysis of complex high-dimensional data, correlate temporal changes in different subsets and lead to biomarker discovery is needed. Here we apply such a method, called cytometric fingerprinting (CF), to data obtained on peripheral blood B cells from an adult patient with type-1 diabetes who underwent pancreatic islet transplantation. We establish that CF can be used to analyze longitudinal trends in immunophenotypic data, and show that results from CF are comparable to those obtained with traditional gating methods. Both methods reveal the appearance of transitional B cells and subsequent accumulation of more mature B cells following immunosuppression and transplantation. This pattern is consistent with a temporally ordered process of B cell auto-reconstitution. We also show the comparative efficiency of fingerprinting in recognizing relative changes in B cell subsets with respect to time, its ability to couple the data with statistical methods (agglomerative clustering) and its potential to define novel subsets.     

血清蛋白质指纹图谱诊断模型在早期胰腺癌中的应用研究

[目的]利用表面增强激光解吸电离飞行时间质谱（SELDI-TOF-MS）技术和生物信息学方法筛选胰腺癌的血清肿瘤标志物。[方法]用弱阳离子交换芯片（CM10）结合并用SELDI-TOF-MS检测54例胰腺癌样本（22例早期胰腺癌,32例晚期胰腺癌）。用支持向量机（SVM）方法建立辨别模型。[结果]利用筛选出的5个蛋白峰（m/z6667.68、8572.38、2958.76、6441.59、5913.36Da）用于建立区分早、晚期胰腺癌的SVM模型,模型的灵敏度和特异性分别为90.9%和78.1%。[结论]SELDI-TOF-MS技术结合生物信息学方法可找到辨别早期胰腺癌和晚期胰腺癌的标志物,并建立区分模型。