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《Drug metabolism and pharmacokinetics》2019,34(5):308-316
LC-MS quantification of drug metabolites is sometimes impeded by the availability of internal standards that often requires customized synthesis and/or extensive purification. Although isotopically labeled internal standards are considered ideal for LC-MS/MS based quantification, de novo synthesis using costly isotope-enriched starting materials makes it impractical for early stage of drug discovery. Therefore, quick access to these isotope-enriched compounds without chemical derivatization and purification will greatly facilitate LC-MS/MS based quantification. Herein, we report a novel 18O-labeling technique using metabolizing enzyme carboxylesterase (CES) and its potential application in metabolites quantification study. Substrates of CES typically undergo a two-step oxygen exchange with H218O in the presence of the enzyme, generating singly- and doubly-18O-labeled carboxylic acids; however, unexpected hydrolytic behavior was observed for three of the test compounds – indomethacin, piperacillin and clopidogrel. These unusual observations led to the discovery of several novel hydrolytic mechanisms. Finally, when used as internal standard for LC-MS/MS based quantification, these in situ labeled compounds generated accurate quantitation comparable to the conventional standard curve method. The preliminary results suggest that this method has potential to eliminate laborious chemical synthesis of isotope-labeled internal standards for carboxylic acid-containing compounds, and can be developed to facilitate quantitative analysis in early-stage drug discovery. 相似文献
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《Clinical neurophysiology》2020,131(1):213-224
ObjectiveSystematically review the abnormalities in event related potential (ERP) recorded in Rett Syndrome (RTT) patients and animals in search of translational biomarkers of deficits related to the particular neurophysiological processes of known genetic origin (MECP2 mutations).MethodsPubmed, ISI Web of Knowledge and BIORXIV were searched for the relevant articles according to PRISMA standards.ResultsERP components are generally delayed across all sensory modalities both in RTT patients and its animal model, while findings on ERPs amplitude strongly depend on stimulus properties and presentation rate. Studies on RTT animal models uncovered the abnormalities in the excitatory and inhibitory transmission as critical mechanisms underlying the ERPs changes, but showed that even similar ERP alterations in auditory and visual domains have a diverse neural basis. A range of novel approaches has been developed in animal studies bringing along the meaningful neurophysiological interpretation of ERP measures in RTT patients.ConclusionsWhile there is a clear evidence for sensory ERPs abnormalities in RTT, to further advance the field there is a need in a large-scale ERP studies with the functionally-relevant experimental paradigms.SignificanceThe review provides insights into domain-specific neural basis of the ERP abnormalities and promotes clinical application of the ERP measures as the non-invasive functional biomarkers of RTT pathophysiology. 相似文献
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目的 观察电针对狗幽门压力、胃黏膜血流量的调控作用及其与血浆、胃黏膜组织中一氧化氮 (NO) ,一氧化氮合酶 (NOS)水平变化的关系 ,以探讨电针对胃黏膜保护作用的机制。方法 将 2 0只狗随机分为 4组 :空白对照组、非经非穴组、上巨虚组、足三里组 (每组 5只 )。采用胃压测量仪、激光多普勒血流仪监测幽门压力、频率及胃黏膜血流量的变化 ,同步测定血浆及胃黏膜组织中NO ,NOS含量 ,并观察变化规律。结果 电针后足三里组幽门括约肌总压力、基础压下降、频率下降 (P <0 .0 5 ) ,胃黏膜血流量显著升高 (4 .5 1± 0 .73→ 6.90± 1.0 1,P <0 .0 1) ,血浆及胃黏膜组织中NO ,NOS含量显著升高 (P <0 .0 5 ) ,上巨虚组仅幽门括约肌总压力下降 ,血浆NO含量上升。但足三里组各项监测指标变化趋势更明显 ,其他组变化无显著性。结论 电针可使狗幽门压力、频率下降 ,使胃黏膜血流量增加 ,与影响幽门压力、胃黏膜血流量的某些活性物质的含量改变有关 ,并具有一定的经络和穴位特异性。 相似文献
16.
作者用自制的[~(125)I]标记激素,参考国外经验,建立了人血清胰岛素抗体、猪胰岛素原抗体和胰多肽抗体等三种放射免疫检测法,并对方法的主要实验条件进行了优选,对方法的质量控制参数作了验证。结果发现:在使用国产胰岛素的糖尿病人血清中,三种抗体的检出率分别为90.8%、48.3%和36.5%;抗体特异性结合值分别为21.2±17.4%、41.8±27.4%和25.6±28.4;而正常人和糖尿病人未用胰岛素者全部为阴性,显示国产胰岛素具有明显的免疫原性。 相似文献
17.
大鼠皮质神经元的体外培养和纯化 总被引:8,自引:2,他引:6
目的:探讨大鼠皮质神经元分离纯化的有效方法。方法:取胎鼠大脑皮质细胞,分别在不同的培养液中进行原代培养,利用相差显微镜对培养的细胞进行动态观察;并以免疫细胞化学技术对神经元进行染色。结果:在相差显微镜下可见加用N2添加剂和胎牛血清培养的神经元生长良好,胞体形态多样,突起数目多,长度长,与邻近神经元的胞体或突起形成接触,而胶质细胞则大量死亡,神经元纯化率达94%以上,未加用N2添加剂培养的神经元胶质细胞大量存在,神经元纯化率达50%左右。结论:N2添加剂和胎牛血清联合应用可使大鼠皮质神经元得到良好的生长和有效纯化。 相似文献
18.
目的观察含补阳还五汤的药物血清对体外培养大鼠脑皮层神经元生长的影响,为补阳还五汤治疗缺血性脑损伤提供实验依据。方法以体外培养新生SD乳鼠大脑皮层神经元为观察对象,采用四唑盐(MTT)比色法测定细胞生长情况。观察含药血清对常规培养及在缺氧状态下神经元生长的影响。结果补阳还五汤对常规培养及在缺氧条件下培养的神经元有显著促进生长作用,与空白血清组比较有显著差(P<0.05)。结论大鼠以补阳还五汤灌胃给药后,其血清中的药物成份有促进体外培养神经元细胞生长的作用。 相似文献
19.
J. LEAKE S. J. KELLIE† J. PRITCHARD† J. M. CHESSELLS† R. A. RISDON 《Histopathology》1989,14(3):255-268
A review of the pathological material from 42 children with non-Hodgkin's lymphoma seen over a 44 month period revealed 10 large cell tumours. Of these, six were classified as peripheral T-cell lymphoma, an entity rarely reported in childhood. Three patients were boys and three girls (median age 9.5 years), and extranodal presentation was a feature of two patients. Five had high-grade tumours; of these, three were classified as large cell anaplastic, Ki-1 positive and two as pleomorphic large cell. The remaining patient had a low-grade tumour of angioimmunoblastic type. T-cell subsets were examined in three cases and showed the following phenotypes: CD4-, CD8-; CD4+, CD8-; CD4-, CD8+. Three of the patients with high-grade tumours died, with a mean survival of 22 weeks. The remaining patients are alive and clinically disease-free for between 10 and 24 months after treatment. 相似文献
20.
P F Maness 《Journal of neuroscience research》1986,16(1):127-139
The advent of recombinant DNA technology has led to the identification in the DNA of normal animal cells of over 30 proto-oncogenes that are homologous to retroviral transforming genes. One of these encodes a protein kinase (pp60c-src) of unknown function, that is preferentially synthesized in brain and neural retina. Here the expression of pp60c-src in the peripheral nervous system was examined in sensory neurons from chick dorsal root ganglia with antisera raised against the transforming protein of Rous sarcoma virus (pp60v-src) expressed in Escherichia coli carrying the cloned v-src gene. This antiserum recognizes pp60c-src specifically in normal chicken cells. Western immunoblotting showed that dorsal root ganglia of stage 30 (day 6.5) chick embryos contained elevated levels of pp60c-src. Immunoperoxidase staining of neuron-enriched cultures prepared from chick dorsal root ganglia showed pp60c-src immunoreactivity in cells with neuronal morphology; flat, fibroblastic cells contained no detectable immunoreactivity. Indirect double immunofluorescence with pp60src antibodies and monoclonal antibodies against the 200-kD subunit of neurofilament protein confirmed that the cells expressing pp60c-src were neurons. Ninety-six percent of the neurofilament-positive cells were immunoreactive with pp60src antibodies, and conversely, all pp60c-src-positive cells were immunoreactive with neurofilament antibodies. pp60c-src immunofluorescence appeared to be distributed over the cell body, processes, and growth cones. These results clearly demonstrate that pp60c-src is a product of neurons and is expressed in sensory neurons in culture. 相似文献