蛋白质指纹图谱技术探索硒诱导金属硫蛋白对小鼠肺纤维化的防护作用

【目的】利用蛋白质指纹图谱技术探索硒诱导金属硫蛋白（metallothionein,MT）对小鼠肺纤维化的防护作用。【方法】利用表面增强的激光解析离子化-飞行时间-质谱（SELDI-TOF-MS）技术测定硒对MT的诱导,寻找小鼠肺纤维化的异常蛋白表达标志并评估硒代MT的防护作用。【结果】IMAC-Cu芯片表达的蛋白质中发现分子量为6277 Da的一组脱金属MT2蛋白表达增强证实硒对MT的高诱导作用;在WCX2和H4芯片中,发现肺纤维化小鼠中肺组织和血清中存在分子量分别为3880 Da和3 364 Da的两蛋白的表达与正常组比较明显受到抑制,硒诱导MT增高后上述异常表达均有改善,说明有机硒可能对肺纤维化诱导产生的蛋白异常具有一定的防护作用。【结论】利用蛋白质指纹图谱技术分析组织和血清的蛋白特异性表达可以作为肺纤维化疗效监测和评价的一类标志物,为今后进行更多的药物或方法筛选提供了一种可以尝试的实验模式。     

乌拉尔甘草优良品系选育研究(Ⅰ)——4个来源甘草遗传基础的AFLP分析

目的研究4个来源甘草属植物的遗传基础,初步探讨扩增片段长度多态性(AFLP)分子标记技术在甘草品系选育中的应用。方法采用AFLP标记技术对乌拉尔甘草栽培新品系"民勤1号"、"喀什1号"、"阿克苏1号"和常规栽培品系内蒙古乌拉尔甘草进行基因组DNA多态性、指纹图谱和UPGMA聚类分析。结果从64对引物组合中筛选出了8对引物组合进行多态性分析,以多态位点百分率最高的E-AAC/M-CAG组合构建4个来源甘草的指纹图谱。UPGMA聚类分析将4个来源甘草分为4组,表现出不同的亲缘关系。结论"民勤1号"、"喀什1号"、"阿克苏1号"形成了独特的基因构成,可以作为乌拉尔甘草优良栽培新品系选育目标进行深入研究。     

应用表面加强激光解吸电离-飞行时间-质谱技术检测结直肠癌肝转移

目的 寻找与结直肠癌肝转移相关的蛋白质,建立结直肠癌肝转移的血清蛋白质指纹图谱诊断预测模型.方法 应用表面加强激光解吸电离-飞行时间-质谱(SELDI-TOF-MS)技术,对36例结直肠癌无肝转移患者和36例结直肠癌伴肝转移患者的术前空腹外周静脉血标本,进行蛋白质指纹图谱测定,运用Biomarker Wizard软件,建立结直肠癌肝转移的诊断预测模型.用44例结直肠癌患者和44例结直肠癌伴肝转移患者,对所建立的诊断预测模型进行盲法验证.结果 比较36例结直肠癌无肝转移患者和36例结直肠癌伴肝转移患者的血清蛋白质,得到10个差异蛋白峰(P<0.05),质荷比分别为2398、2814、4084、4289、4465、6422、6619、11 482、11 649和13 714.若以P<0.01为标准,则有3个蛋白质峰差异有统计学意义,质荷比分别为2398、2814和13714.建立终末节点数为9的诊断预测模型,其敏感性为91.7%,特异性为97.2%.验证结果显示,敏感性为75.0%,特异性为81.8%.结论 运用SELDI-TOF-MS技术所建立的血清蛋白指纹图谱模型,在预测结直肠癌肝转移中具有非常高的敏感性与特异性,可望成为预测诊断工具.     

First molecular characterisation of Cryptosporidium and Giardia from Bubalus bubalis (water buffalo) in Victoria,Australia

We conducted a molecular epidemiological survey of Cryptosporidium and Giardia from Bubalus bubalis (water buffalo) on two extensive farms (450 km apart) in Victoria, Australia. Faecal samples (n = 476) were collected from different age groups of water buffalo at two time points (six months apart) and tested using a PCR-based mutation scanning-targeted sequencing-phylogenetic approach, employing markers within the small subunit of ribosomal RNA (designated pSSU) and triose phosphate isomerase (ptpi) genes. Based on pSSU data, Cryptosporidium parvum, Cryptosporidium bovis and Cryptosporidium genotypes 1, 2 (each 99% similar genetically to Cryptosporidium ryanae) and 3 (99% similar to Cryptosporidium suis) were detected in two (0.4%), one (0.2%), 38 (8.0%), 16 (3.4%) and one (0.2%) of the 476 samples tested, respectively. Using ptpi, Giardia duodenalis assemblages A and E were detected in totals of 56 (11.8%) and six (1.3%) of these samples, respectively. Cryptosporidium was detected on both farms, whereas Giardia was detected only on farm B, and both genera were detected in 1.5% of all samples tested. The study showed that water buffaloes on these farms excreted C. parvum and/or G. duodenalis assemblage A, which are consistent with those found in humans, inferring that these particular pathogens are of zoonotic significance. Future work should focus on investigating, in a temporal and spatial manner, the prevalence and intensity of such infections in water buffaloes in various geographical regions in Australia and in other countries.     

当归补血、活血作用的谱效关系研究

用乙酰苯肼(APH)复制血虚小鼠模型,于实验第1,4,7 d皮下注射APH,剂量为200,100,100 mg·kg^-1(0.01 mL·g^-1)。于造模第10 天开始灌胃给予当归提取物 (生药剂量为2.4 g·kg^-1),连续10 d。观察当归不同提取部位对血虚小鼠RBC、Hb、PLT、胸腺、脾脏及体重变化等指标的影响。用灰色关联统计方法将HPLC指纹图谱各共有峰的峰面积和补血、活血药效数据相关联,研究谱效相关性。结果显示,水提醇沉上清(DSC)部位补血、活血效果较好。根据关联度大小确定了DSC中各成分对补血、活血作用贡献程度。其中,阿魏酸的贡献最大,但不应忽略其他成分的贡献。该文研究有效组分的HPLC指纹图谱及谱效关系,确定了DSC部分补血、活血的物质基础,为表征其谱效相关性提供有效途径。     

Effects of Postharvest Processing and Geographical Source on Phytochemical Variation of Corydalis Rhizoma

Objective To assess the relative contributions of postharvest processing and geographical source to phytochemical variation of Corydalis Rhizoma, and rhizome of Corydalis yanhusuo, and to examine what phytochemical components are the most sensitive to the differences of each factor and how they change. Methods HPLC fingerprinting and LC-MS coupled with chemometric approaches were applied. Results The results of principal component analysis(PCA) and hierarchical cluster analysis(HCA) explicitly demonstrated the postharvest processing could produce a greater impact on the phytochemical profiles of Corydalis Rhizoma than geographical source. The contents of most compounds increased after water boiling while decreased after sulphur-fumigation.Protopine, coptisine, and palmatine were the most variable components in processing. Geographical sources also led to a remarkable phytochemical differentiation, in which the environmental variation of the three regions might play a role. Dehydrocorybulbine, coptisine, dehydrocorydaline, and protopine varied most among the three production regions and decreased sequentially in Zhejiang, Shaanxi, and Jiangsu provinces, China. Conclusion Both postharvest processing and geographical source should be enhanced with the priority for the former in the quality control of Corydalis Rhizoma. The application of boiling is supported but the consistency should be improved in practice. Sulphur-fumigation is strongly suggested to be abandoned.     

Capillary electrophoresis as a metabolomic tool in antioxidant therapy studies

The development of an approach by which two CE methods operating with opposite polarities and orthogonal capillary electrophoretic separation modes (method 1: normal polarity cyclodextrin modified MEKC (CD-MEKC) and method 2: reversed polarity CZE) for the sequential application to urinary samples from a type I diabetes metabolomics investigation is discussed. During method development, problematic MEKC profile drift issues arising from the high glucose content of the diabetic animal urine samples required some electrolyte modifications involving the use of hexafluoroisopropanol (HFIP) to circumvent the drift. Data derived from both methods were subsequently subjected to alignment, normalization and multi-dimensional scaling (MDS) procedures. In such a way, classification of samples derived from control and diabetic animals receiving a placebo from those receiving an antioxidant nutraceutical, was successfully demonstrated. Such a strategy is a cost effective and comprehensive metabolomics tool useful for describing UV absorbing metabolite disease-related changes in nutra/pharma-ceutical studies.     

新疆维吾尔族肾癌血清蛋白指纹图谱的建立

目的 筛选并建立新疆维吾尔族肾癌血清蛋白指纹图谱诊断模型. 方法 采用弱阳离子交换蛋白质芯片(CM10蛋白芯片)及表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术对45例维吾尔族肾癌患者和45例正常对照者血清指纹图谱进行检测,结果用ZUCI-蛋白芯片数据分析系统软件包进行分析,通过支持向量机运算建立区分肾癌蛋白指纹图谱诊断模型,留一法交叉验证作用评估模型,判别效果. 结果 两组血清中筛选出M/Z为4296、4305、5914、5935、6116、6887、8085、8142、8573共9个差异有统计学意义(P＜0.05)的标志蛋白,所建立的诊断模型诊断肾癌的灵敏性为100％ (45/45),特异性为91％ (41/45).进一步用50例未知血清标本盲法测试该模型,双盲验证后的灵敏性和特异性分别为93％ (28/30)和85％ (17/20). 结论 SELDI-TOF-MS结合支持向量机建立维吾尔族肾癌血清蛋白质指纹图谱模型具有较高的敏感性与特异性,值得进一步研究和应用.     

Cortical responses to noninvasive perturbations enable individual brain fingerprinting

BackgroundIn recent years, it has become increasingly apparent that characterizing individual brain structure, connectivity and dynamics is essential for understanding brain function in health and disease. However, the majority of neuroimaging and brain stimulation research has characterized human brain function by averaging measurements from groups of subjects and providing population-level inferences. External perturbations applied directly to well-defined brain regions can reveal distinctive information about the state, connectivity and dynamics of the human brain at the individual level.ObjectivesIn a series of studies, we aimed to characterize individual brain responses to MRI-guided transcranial magnetic stimulation (TMS), and explore the reproducibility of the evoked effects, differences between brain regions, and their individual specificity.MethodsIn the first study, we administered single pulses of TMS to both anatomically (left dorsolateral prefrontal cortex- ‘L-DLPFC’, left Intra-parietal lobule- ‘L-IPL) and functionally (left motor cortex- ‘L-M1’, right default mode network- ‘R-DMN, right dorsal attention network- ‘R-DAN’) defined cortical nodes in the frontal, motor, and parietal regions across two identical sessions spaced one month apart in 24 healthy volunteers. In the second study, we extended our analyses to two independent data sets (n = 10 in both data sets) having different sham-TMS protocols.ResultsIn the first study, we found that perturbation-induced cortical propagation patterns are heterogeneous across individuals but highly reproducible within individuals, specific to the stimulated region, and distinct from spontaneous activity. Most importantly, we demonstrate that by assessing the spatiotemporal characteristics of TMS-induced brain responses originating from different cortical regions, individual subjects can be identified with perfect accuracy. In the second study, we demonstrated that subject specificity of TEPs is generalizable across independent data sets and distinct from non-transcranial neural responses evoked by sham-TMS protocols.ConclusionsPerturbation-induced brain responses reveal unique “brain fingerprints” that reflect causal connectivity dynamics of the stimulated brain regions, and may serve as reliable biomarkers of individual brain function.