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Correction: Rapidly improved determination of metabolites from biological data sets using the high-efficient TransOmics tool.
Mol Biosyst
PUBLISHED: 11-20-2014
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Correction for 'Rapidly improved determination of metabolites from biological data sets using the high-efficient TransOmics tool' by Aihua Zhang et al., Mol. BioSyst., 2014, 10, 2160-2165.
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Controllably Interfacing with Metal: A Strategy for Enhancing CO Oxidation on Oxide Catalysts by Surface Polarization.
J. Am. Chem. Soc.
PUBLISHED: 10-13-2014
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Heterogeneous catalysis often involves charge transfer from catalyst surface to adsorbed molecules, whose activity thus depends on the surface charge density of catalysts. Here, we demonstrate a unique solution-phase approach to achieve controllable interfacial lengths in oxide-metal hybrid structures. Resulting from their different work functions, surface polarization is induced by the Ag-CuO interface and acts to tailor the surface charge state of CuO. As a result, the designed hybrid catalysts exhibit enhanced intrinsic activities in catalyzing CO oxidation in terms of apparent activation energy, as compared with their counterparts. Moreover, the CO conversion rate can be enhanced by maximizing the Ag-CuO interfacial length and thus the number of active sites on the CuO. This work provides a new strategy for tuning catalytic performance by controlling interface in hybrid catalysts.
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Urinary metabolic profiling of rat models revealed protective function of scoparone against alcohol induced hepatotoxicity.
Sci Rep
PUBLISHED: 09-29-2014
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Alcohol-induced liver disease (ALD) is a leading cause of non-accident-related deaths in the world. Identification of an early specific signature of ALD would aid in therapeutic intervention. Scoparone is an important constituent of Yinchenhao, and displayed bright prospects in hepatoprotective effect. However, its precise molecular mechanism has not been well explored. The present study was designed to assess the effects and possible mechanisms of scoparone against alcohol-induced liver injury. UPLC/ESI-Q-TOF/MS combined with pattern recognition approaches including PCA, and PLS-DA were integrated to get differentiating metabolites for the pathways and clarify mechanisms of disease, highlight insights into drug discovery. The results indicated four ions in the positive mode were characterized as potential differentiating metabolites which can be regulated by scoparone treatment, and suggested that therapeutic effect of scoparone could regulated the dysfunctions of citrate cycle, sphingolipid metabolism, taurine and hypotaurine.
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Rapidly improved determination of metabolites from biological data sets using the high-efficient TransOmics tool.
Mol Biosyst
PUBLISHED: 06-04-2014
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Metabolomics is a new approach based on the systematic study of the full complement of metabolites in a biological sample. Extracting biomedical information from large datasets is of considerable complexity. Furthermore, the traditional method of assessing metabolomics data is not only time-consuming but it is often subjective work. Here we used sensitive ultra-performance LC-ESI/Q-TOF high-definition mass spectrometry (UPLC-ESI-Q-TOF-MS) in positive ion mode coupled with a new developed software program TransOmics for widely untargeted metabolomics, which incorporates novel nonlinear alignment, deconvolution, matched filtration, peak detection, and peak matching to characterize metabolites as a case study. The TransOmics method can facilitate prioritization of the data and greatly increase the probability of identifying metabolites related to the phenotype of interest. By this means, 17 urinary differential metabolites were identified (less than 10 min) involving the key metabolic pathways including tyrosine metabolism, glutathione metabolism, phenylalanine metabolism, ascorbate and aldarate metabolism, arginine and proline metabolism, and so forth. Metabolite identification has also been significantly improved, using the correlation peak patterns in contrast to a reference metabolite panel. It can detect and identify metabolites automatically and remove background noise, and also provides a user-friendly graphical interface to apply principal component analyses, correlation analysis and compound statistics. This investigation illustrates that metabolomics combined with the proposed bioinformatic approach (based on TransOmics) is important to elucidate the developing biomarkers and the physiological mechanism of disease, and has opened the door for the development of a new genre of metabolite identification methods.
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Fingerprinting and Simultaneous Determination of Alkaloids and Limonins in Phellodendri Amurensis Cortex From Different Locations by High-Performance Liquid Chromatography with Diode Array Detection.
J Chromatogr Sci
PUBLISHED: 05-30-2014
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A sensitive high-performance liquid chromatography method coupled with diode array detection (HPLC-DAD) was developed for the quality control of Phellodendri amurensis cortex (PAC), the quality control included the simultaneous determination of seven major constituents, namely phellodendrine, magnoflorine, jatrorrhizine, palmatine, berberine, obaculactone and obacunone. The chromatographic separation was accomplished on a Diamonsil-C18 column (4.6 mm × 200 mm, 5 ?m) with acetonitrile and 0.1% phosphoric acid (0.02 mol sodium dihydrogen phosphate per liter) by linear gradient elution. The established method was successfully validated by acceptable linearity, limits of detection and quantitation, precision, repeatability, stability and accuracy. The HPLC-DAD fingerprint chromatograph under 220 nm consisting of 21 peaks was constructed for the evaluation of the 11 batches of PAC. The HPLC fingerprints were analyzed by similarity analysis, hierarchical clustering analysis and principal component analysis. The results indicated that the combination of multicomponent determination method and chromatographic fingerprint analysis could be employed for the quantitative analysis and identification of PAC, as well as pharmaceutical products containing this herbal material.
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Induction of protective immune response against both PPRV and FMDV by a novel recombinant PPRV expressing FMDV VP1.
Vet. Res.
PUBLISHED: 05-27-2014
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Peste des petits ruminants (PPR) and foot-and-mouth disease (FMD) are both highly contagious diseases of small domestic and wild ruminants caused by the PPR virus (PPRV) and the FMD virus (FMDV). In this study, a recombinant PPRV expressing the FMDV VP1 gene (rPPRV/VP1) was generated and FMDV VP1 expression did not impair replication of the recombinant virus in vitro and immunogenicity in inducing neutralizing antibody against PPR in goats. Vaccination with one dose of rPPRV/VP1 induced FMDV neutralizing antibody in goats and protected them from challenge with virulent FMDV. Our results suggest that the recombinant PPRV expressing the FMDV VP1 protein is a potential dual live vectored vaccine against PPRV and FMDV.
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Metabolomics approaches and applications in prostate cancer research.
Appl. Biochem. Biotechnol.
PUBLISHED: 05-09-2014
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Prostate cancer is a leading cause of cancer deaths in men worldwide. Although prostate-specific antigen (PSA) has been extensively used as a serum biomarker to detect prostate cancer, this screening method has suffered from a lack of specificities and sensitivities. The successful prevention and treatment of prostate cancer relies on the early and accurate detection of the disease; therefore, more sensitive biomarkers are urgently needed. Prostate has long been known to exhibit unique metabolite profiles, fortunately, metabolomics, the study of all metabolites produced in the body, can be considered most closely related to a patient's phenotype. It may provide clinically useful biomarkers applied toward identifying metabolic alterations in prostate cancer and has introduced new insights into the pathology of prostate cancer. This advanced bioanalytic method may now open door for diagnostics. Metabolomics has a great and largely potential in the field of disease, and the analysis of the cancer metabolome to identify novel biomarkers and targets can now be undertaken in many research laboratories. In this review, we take a closer look at the metabolomics in the field of prostate cancer and highlight the interesting publications as references for the interested reader.
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Two-dimensional g-C(3)N(4): an ideal platform for examining facet selectivity of metal co-catalysts in photocatalysis.
Chem. Commun. (Camb.)
PUBLISHED: 04-30-2014
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Two-dimensional g-C3N4 nanosheets with few-layer thickness, ensuring equivalent charge migrations to various Pd facets, provide an ideal model system for reliably examining the facet selectivity of Pd co-catalysts. It reveals that reduction of CO2 can occur better on Pd{111} facets while H2O prefers to generate H2 on Pd{100}.
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A caryophyllane-type sesquiterpene, caryophyllenol A from Valeriana amurensis.
Fitoterapia
PUBLISHED: 03-25-2014
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A new caryophyllane derivative, caryophyllenol A, and a new germacrane derivative, isovolvalerenal D, together with 11 known sesquiterpenoids, were isolated from a petroleum ether partition of the roots and rhizomes of Valeriana amurensis. Structure elucidation of caryophyllenol A and isovolvalerenal D was accomplished on the basis of various spectroscopic techniques including HRESIMS and 2D NMR analyses. The structure of caryophyllenol A was further confirmed by X-ray crystallography and using quantum-chemical ECD calculation adopting TDDFT method. Caryophyllenol A and other eight sesquiterpenoids were evaluated for sedative activity with the model of Drosophila melanogaster, and eight of them showed the effect of prolonging the total sleeping time (TST) of D. melanogaster, displaying significant sedative action.
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Identification and characterization of the chemical constituents of Simiao Wan by ultra high performance liquid chromatography with mass spectrometry coupled to an automated multiple data processing method.
J Sep Sci
PUBLISHED: 01-31-2014
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The chemical constituents of Simiao Wan (SW), a traditional Chinese medicine preparation, are difficult to determine and remain unclear. To more efficiently detect ions, a multiple data processing approach has been used in the characterization of the compounds. In this study, a rapid and sensitive method based on ultra high performance liquid chromatography with mass spectrometry and the multiple data processing approach was established to characterize the chemical constituents of SW. Ultra high performance liquid chromatography with mass spectrometry coupled with the multiple data processing approach could efficiently remove nonrelated ion signals from accurate mass data. We report the application of the multiple data processing approach for comprehensive detection and rapid identification of chemical constituents of SW. Of note, the total analysis time for separation was less than 20 min without losing any resolution. In the variable, importance in projection plot of orthogonal projection to latent structure-discriminant analysis, a total of 72 ions of interest (37 ions in positive mode, 38 ions in negative mode and three ions in both mode) were extracted or tentatively characterized based on their retention times, exact mass measurement for each molecular ion and subsequent fragment ions. In summary, the methodology proposed in this study could be valuable for the structural characterization and identification of the multiple constituents in the traditional Chinese medicine formula SW.
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The application of metabolomics in traditional Chinese medicine opens up a dialogue between Chinese and Western medicine.
Phytother Res
PUBLISHED: 01-30-2014
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Metabolomics provides an opportunity to develop the systematic analysis of the metabolites and has been applied to discovering biomarkers and perturbed pathways which can clarify the action mechanism of traditional Chinese medicines (TCM). TCM is a comprehensive system of medical practice that has been used to diagnose, treat and prevent illnesses more than 3000?years. Metabolomics represents a powerful approach that provides a dynamic picture of the phenotype of biosystems through the study of endogenous metabolites, and its methods resemble those of TCM. Recently, metabolomics tools have been used for facilitating interactional effects of both Western medicine and TCM. We describe a protocol for investigating how metabolomics can be used to open up 'dialogue' between Chinese and Western medicine, and facilitate lead compound discovery and development from TCM. Metabolomics will bridge the cultural gap between TCM and Western medicine and improve development of integrative medicine, and maximally benefiting the human. Copyright © 2014 John Wiley & Sons, Ltd.
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Designing p-type semiconductor-metal hybrid structures for improved photocatalysis.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 01-29-2014
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A practical strategy is proposed to facilitate the migration of holes in semiconductor (the low rate of which limits photocatalytic efficiency) by taking advantage of the Schottky barrier between p-type semiconductor and metal. A high work function is found to serve as an important selection rule for building such desirable Schottky junction between semiconductor surface facets and metal. The intrinsic charge spatial distribution has to be taken into account when selecting the facets, as it results in accumulation of photoexcited electrons and holes on certain semiconductor facets. Importantly, the facets have a high work function, the same characteristic required for the formation of Schottky junction in a p-type semiconductor-metal hybrid structure. As a result, the semiconductor crystals in the hybrid design may be better enclosed by single facets with high work function, so as to synergize the two effects: Schottky barrier versus charge spatial separation.
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Integration of an inorganic semiconductor with a metal-organic framework: a platform for enhanced gaseous photocatalytic reactions.
Adv. Mater. Weinheim
PUBLISHED: 01-27-2014
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Ultrafast spectroscopy demonstrates that charge transfer can occur between photoexcited inorganic semiconductors and metal-organic frameworks (MOFs), supplying long-lifetime electrons for the reduction of gas molecules adsorbed on the MOF. As a proof of concept, a unique method is developed for synthesizing Cu3 (BTC)2 @TiO2 core-shell structures with macroporous semiconductor shells that allow gas molecules to be captured in the cores.
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Ultra-performance liquid chromatography tandem mass spectrometry combined with automated MetaboLynx analysis approach to screen the bioactive components and their metabolites in Wen-Xin-Formula.
Biomed. Chromatogr.
PUBLISHED: 01-08-2014
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Wen-Xin-Formula (WXF), a famous traditional prescription, has been widely used to treat myocardial ischemia syndrome for thousands of years. However, the constituents absorbed into blood after oral administration of WXF remain unknown. Here, an integrative ultra performance liquid chromatography coupled with electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS) combined with the MetaboLynx approach was established to investigate the absorbed constituents in rats after oral administration of WXF. A hyphenated electrospray ionization and quadrupole-time-of-flight analyzer was used for the determination of accurate mass of the molecule and fragment ions. With this rapid and automated analysis method, a total of 32 peaks were tentatively characterized in vivo based on MS and MS/MS data and comparison with available databasess, 26 of which were parent components and six metabolites. These components mainly were ginsenosides, paeoniflorin, galloyl glucose, berberis alkaloids, phenolic, phenolic glycosides and unsaturated fatty acids, glucuronide products of original berberis alkaloids. The present study demonstrates that integrative UPLC-ESI-Q-TOF-MS technique and MetaboLynx data processing method were successfully applied for the rapid discovery of potentially bioactive components and metabolites from WXF, and proved that the established method could help to explore the effective substances for further research into WXF. Copyright © 2014 John Wiley & Sons, Ltd.
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Metabolomic analysis of diet-induced type 2 diabetes using UPLC/MS integrated with pattern recognition approach.
PLoS ONE
PUBLISHED: 01-01-2014
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Metabolomics represents an emerging discipline concerned with comprehensive assessment of small molecule endogenous metabolites in biological systems and provides a powerful approach insight into the mechanisms of diseases. Type 2 diabetes (T2D), called the burden of the 21st century, is growing with an epidemic rate. However, its precise molecular mechanism has not been comprehensively explored. In this study, we applied urinary metabolomics based on the UPLC/MS integrated with pattern recognition approaches to discover differentiating metabolites, to characterize and explore metabolic pathway disruption in an experimental model for high-fat-diet induced T2D. Six differentiating urinary metabolites were found in the negative mode, and two (2-(4-hydroxy-3-methoxy-phenyl) acetaldehyde sulfate, 2-phenylethanol glucuronide) of which were identified involving the key metabolic pathways linked to pentose and glucuronate interconversions, starch, sucrose metabolism and tyrosine metabolism. Our study provides new insight into pathophysiologic mechanisms and may enhance the understanding of T2D pathogenesis.
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Novel in-ovo chimeric recombinant Newcastle disease vaccine protects against both Newcastle disease and infectious bursal disease.
Vaccine
PUBLISHED: 01-01-2014
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Development of a safe and efficient in-ovo vaccine against Newcastle disease (NDV) and very virulent infectious bursal disease virus (vvIBDV) is of great importance. In this study, a chimeric NDV LaSota virus with the L gene of Clone-30 (rLaC30L) was used to generate a recombinant chimeric virus expressing the VP2 protein of vvIBDV (rLaC30L-VP2). The safety and efficacy of rLaC30L-VP2 in-ovo vaccination was then evaluated in 18-day-old special pathogen free (SPF) chicken embryos and commercial broiler embryos for prevention of NDV and vvIBDV. Hatchability and global survival rate of the hatched birds was not affected by in-ovo rLaC30L-VP2 vaccination. However, rLaC30L-VP2 in-ovo vaccination induced significant anti-IBDV and anti-NDV antibodies in SPF birds and commercial broilers, and 100% of vaccinated chickens were protected against a lethal NDV challenge. In-ovo rLaC30L-VP2 vaccination also provided resistance against vvIBDV challenge in a significant amount of animals. These results suggest that rLaC30L-VP2 is a safe and efficient bivalent live in-ovo vaccine against NDV and vvIBDV.
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Metabolomics in diagnosis and biomarker discovery of colorectal cancer.
Cancer Lett.
PUBLISHED: 11-06-2013
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Colorectal cancer (CRC), a major public health concern, is the second leading cause of cancer death in developed countries. There is a need for better preventive strategies to improve the patient outcome that is substantially influenced by cancer stage at the time of diagnosis. Patients with early stage colorectal have a significant higher 5-year survival rates compared to patients diagnosed at late stage. Although traditional colonoscopy remains the effective means to diagnose CRC, this approach generally suffers from poor patient compliance. Thus, it is important to develop more effective methods for early diagnosis of this disease process, also there is an urgent need for biomarkers to diagnose CRC, assess disease severity, and prognosticate course. Increasing availability of high-throughput methodologies open up new possibilities for screening new potential candidates for identifying biomarkers. Fortunately, metabolomics, the study of all metabolites produced in the body, considered most closely related to a patients phenotype, can provide clinically useful biomarkers applied in CRC, and may now open new avenues for diagnostics. It has a largely untapped potential in the field of oncology, through the analysis of the cancer metabolome to identify marker metabolites defined here as surrogate indicators of physiological or pathophysiological states. In this review we take a closer look at the metabolomics used within the field of colorectal cancer. Further, we highlight the most interesting metabolomics publications and discuss these in detail; additional studies are mentioned as a reference for the interested reader.
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Labeling Thiols on Proteins, Living Cells, and Tissues with Enhanced Emission Induced by FRET.
Sci Rep
PUBLISHED: 10-28-2013
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Using N-(2-Aminoethyl)maleimide-cysteine(StBu) (Mal-Cys) as a medium, protein thiols were converted into N-terminal cysteines. After a biocompatible condensation reaction between the N-terminal cysteine and fluorescent probe 2-cyanobenzothiazole-Gly-Gly-Gly-fluorescein isothiocyanate (CBT-GGG-FITC), a new fluorogenic structure Luciferin-GGG-FITC was obtained. The latter exhibits near one order of magnitude (7 folds) enhanced fluorescence emission compared to the precursor moiety due to fluorescence resonance energy transfer (FRET) effect between the newly formed luciferin structure and the FITC motif. Theoretical investigations revealed the underlying mechanism that satisfactorily explained the experimental results. With this method, enhanced fluorescence imaging of thiols on proteins, outer membranes of living cells, translocation of membrane proteins, and endothelial cell layers of small arteries was successfully achieved.
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Cell metabolomics.
OMICS
PUBLISHED: 08-29-2013
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Abstract Metabolomics technologies enable the examination and identification of endogenous biochemical reaction products, revealing information on the precise metabolic pathways and processes within a living cell. Metabolism is either directly or indirectly involved with every aspect of cell function, and metabolomics is thus believed to be a reflection of the phenotype of any cell. Metabolomics analysis of cells has many potential applications and advantages compared to currently used methods in the postgenomics era. Cell metabolomics is an emerging field that addresses fundamental biological questions and allows one to observe metabolic phenomena in cells. Cell metabolomics consists of four sequential steps: (a) sample preparation and extraction, (b) metabolic profiles of low-weight metabolites based on MS or NMR spectroscopy techniques, (c) pattern recognition approaches and bioinformatics data analysis, (d) metabolites identification resulting in putative biomarkers and molecular targets. The biomarkers are eventually placed in metabolic networks to provide insight on the cellular biochemical phenomena. This article analyzes the recent developments in use of metabolomics to characterize and interpret the cellular metabolome in a wide range of pathophysiological and clinical contexts, and the putative roles of the endogenous small molecule metabolites in this new frontier of postgenomics biology and systems medicine.
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New analytical method for the study of the metabolism of gentiopicroside in rats after oral administration by LC-TOF-MS following picolinoyl derivatization.
J Sep Sci
PUBLISHED: 08-13-2013
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The metabolism of gentiopicroside (GPS) in vivo was studied for the first time by LC-MS following picolinoyl derivatization. Incubation of erythrocentaurin, one of the main in vitro metabolites of GPS by intestinal bacteria, with liver microsome indicated that GPS might be metabolized to a final metabolite 3,4-dihydro-5-(hydroxymethyl)isochroman-1-one (HMIO) in vivo. After hydrolysis with sulfatase, HMIO was successfully detected in rat plasma after oral administration of GPS by LC-MS following picolinoyl derivatization. 4-Methoxyphenyl methanol was used as an internal standard to quantify HMIO in rat plasma. A metabolic pathway of GPS in rats is proposed. The monoterpene compound GPS was found to be metabolized to dihydroisocoumarin, which may be responsible for the pharmacological effect of GPS.
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Highly sensitive determination of new metabolite in rat plasma after oral administration of swertiamarin by liquid chromatography/time of flight mass spectrometry following picolinoyl derivatization.
Biomed. Chromatogr.
PUBLISHED: 08-13-2013
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The metabolism of swertiamarin (STM) in vivo was studied by LC/MS following picolinoyl derivatization. Incubation of erythrocentaurin (ECR), one of the main in vitro metabolites of STM by intestinal bacteria, with liver microsome indicated that STM may be metabolized to the final metabolite 3,4-dihydro-5-(hydroxymethyl) isochroman-1-one (HMIO) in vivo. After hydrolyzation with sulfatase, HMIO was successfully detected in rat plasma after oral administration of STM by LC/MS following picolinoyl derivatization. 4-Methoxyphenyl methanol was used as the internal standard to quantify HMIO in rat plasma. The full metabolic pathway of STM in rats is proposed. STM is first hydrolyzed by bacterial ?-glucusidase to give aglycone, which is readily converted to ECR and nitrogen-containing metabolite. ECR is further reduced to HMIO by both liver and intestinal bacteria and HMIO is finally converted to the new sulfate conjugate metabolite. The monoterpene compound STM was found to be metabolized to dihydroisocoumarin and alkaloid compounds in vivo, which may be responsible for the pharmacological effect of STM. The results may shed light on clinical efficacy of STM and the new analytical method developed may assist in studies of the metabolism of other natural iridoids and secoiridoids in vivo. Copyright © 2013 John Wiley & Sons, Ltd.
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Metabolite profiling and pathway analysis of acute hepatitis rats by UPLC-ESI MS combined with pattern recognition methods.
Liver Int.
PUBLISHED: 07-28-2013
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Metabolomics is comprehensive analysis of low-molecular-weight endogenous metabolites in a biological sample. It could enable mapping of perturbations of early biochemical changes in diseases and hence provide an opportunity to develop predictive biomarkers that could provide valuable insights into the mechanisms of diseases. The aim of this study was to elucidate the changes in endogenous metabolites and to phenotype the metabolic profiling of d-galactosamine (GalN)-inducing acute hepatitis in rats by UPLC-ESI MS.
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Rapid identification and comparative analysis of the chemical constituents and metabolites of Phellodendri amurensis cortex and Zhibai dihuang pill by ultra-performance liquid chromatography with quadrupole TOF-MS.
J Sep Sci
PUBLISHED: 07-21-2013
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Zhibai dihuang pill is a famous traditional Chinese medicine formula containing the herbal medicine Phellodendri amurensis cortex. In this work, an integrative pattern recognition approach including principal component analysis and orthogonal partial least squared discriminant analysis based on ultra-performance liquid chromatography coupled to MS was successfully applied for the rapid discovery of natural compounds from herbal medicines. In a 24 min analysis, 93 compounds were identified or tentatively characterized from Zhibai dihuang pill based on their fragmentation behaviors, 23 of which were from Phellodendri amurensis cortex. A total of 26 metabolites in plasma were identified from Phellodendri amurensis cortex in Zhibai dihuang pill formula, among them, 12 prototypes and 14 metabolites through metabolic pathways of demethylation, methylation, hydrolysis, sulfate conjugation, and glucuronide conjugation, which were seven metabolites more than that of the single drug, suggesting the importance of the compatibility of traditional Chinese medicine. The present study provided important structural information on the metabolism of Zhibai dihuang pill. Furthermore, the results of this work have demonstrated the feasibility of ultra-performance liquid chromatography coupled to MS for the rapid and reliable characterization of metabolites from herbal medicines. Based on these results, this method could be a novel approach for identifying potentially bioactive components in other traditional Chinese medicines.
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Multifunctional fluorescent probe for sequential detections of glutathione and caspase-3 in vitro and in cells.
Anal. Chem.
PUBLISHED: 06-21-2013
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Herein, we report a new "On-On" strategy based on the assembly and disassembly of fluorescein isothiocyanate nanoparticles (FITC-NPs) for sequential detections of glutathione (GSH) and caspase-3 (Casp3) with a multifunctional fluorescent probe 1. Theoretical investigations revealed the underlying mechanism that satisfactorily explained experimental results of such consecutive enhancements of fluorescence. Using this probe, we also successfully imaged the Casp3 activity in apoptotic cells.
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Metabolomics insights into pathophysiological mechanisms of nephrology.
Int Urol Nephrol
PUBLISHED: 06-04-2013
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Kidney diseases (KD), a major public health problem that affects about 10 % of the general population, manifest in progressive loss of renal function, which ultimately leads to complete kidney failure. However, current approaches based on renal histopathological results and clinical parameters lack sensitivity and are not sufficient to characterize the category and progression of nephrology or to predict nephrology progression risk reliably or to guide preventive interventions. The high incidence and financial burden of KD make it imperative to diagnose KD at early stages when therapeutic interventions are far more effective. Nowadays, the appearance of metabolomics (the high-throughput measurement and analysis of metabolites) has provided the framework for a comprehensive analysis of KD and serves as a starting point for generating novel molecular diagnostic tools for use in nephrology. Changes in the concentration profiles of a number of small-molecule metabolites found in either blood or urine can be used to localize kidney damage or assess kidneys suffering from injury. The power of metabolomics allows unparalleled opportunity to query the molecular mechanisms of KD. Novel metabolomics technologies have the ability to provide a deeper understanding of the disease beyond classical histopathology, redefine the characteristics of the disease state, and identify novel approaches to reduce renal failure. This review gives an overview of its application to important areas in clinical nephrology, with a particular focus on biomarker discovery. Great strides forward are being made in breaking down important barriers to the successful prevention and treatment of this devastating disorder.
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Potentiating Therapeutic Effects by Enhancing Synergism Based on Active Constituents from Traditional Medicine.
Phytother Res
PUBLISHED: 05-22-2013
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Shifting current drug discovery tide from finding new drugs to screening natural products may be helpful for overcoming the more investment, fewer drugs challenge. Traditional Chinese medicine (TCM), relying on natural products, has been playing a very important role in health protection and disease control for thousands of years in Asia, whose therapeutic efficacy is based on the synergism, that is, the combinational effects to be greater than that of the individual drug. Based on syndromes and patient characteristics and guided by the theories of TCM, formulae are designed to contain a combination of various kinds of crude drugs that, when combined, generally assume that a synergism of all ingredients will bring about the maximum of therapeutic efficacy. The increasing evidence has shown that multiple active component combinations of TCM could amplify the therapeutic efficacy of each agent, representing a new trend for modern medicine. However, the precise mechanism of synergistic action remains poorly understood. The present review highlights the concept of synergy and gives some examples of synergistic effects of TCM, and provides an overview of the recent and potential developments of advancing drug discovery towards more agile development of targeted combination therapies from TCM. Copyright © 2013 John Wiley & Sons, Ltd.
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Dissection of Biological Property of Chinese Acupuncture Point Zusanli Based on Long-Term Treatment via Modulating Multiple Metabolic Pathways.
Evid Based Complement Alternat Med
PUBLISHED: 05-20-2013
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Acupuncture has a history of over 3000 years and is a traditional Chinese medical therapy that uses hair-thin metal needles to puncture the skin at specific points on the body to promote wellbeing, while its molecular mechanism and ideal biological pathways are still not clear. High-throughput metabolomics is the global assessment of endogenous metabolites within a biologic system and can potentially provide a more accurate snap shot of the actual physiological state. We hypothesize that acupuncture-treated human would produce unique characterization of metabolic phenotypes. In this study, UPLC/ESI-HDMS coupled with pattern recognition methods and system analysis were carried out to investigate the mechanism and metabolite biomarkers for acupuncture treatment at "Zusanli" acupoint (ST-36) as a case study. The top 5 canonical pathways including alpha-linolenic acid metabolism, d-glutamine and d-glutamate metabolism, citrate cycle, alanine, aspartate, and glutamate metabolism, and vitamin B6 metabolism pathways were acutely perturbed, and 53 differential metabolites were identified by chemical profiling and may be useful to clarify the physiological basis and mechanism of ST-36. More importantly, network construction has led to the integration of metabolites associated with the multiple perturbation pathways. Urine metabolic profiling might be a promising method to investigate the molecular mechanism of acupuncture.
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Advancing Drug Discovery and Development from Active Constituents of Yinchenhao Tang, a Famous Traditional Chinese Medicine Formula.
Evid Based Complement Alternat Med
PUBLISHED: 05-09-2013
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Traditional Chinese medicine (TCM) formula has been playing a very important role in health protection and disease control for thousands of years. Guided by TCM syndrome theories, formula are designed to contain a combination of various kinds of crude drugs that, when combined, will achieve synergistic efficacy. However, the precise mechanism of synergistic action remains poorly understood. One example is a famous TCM formula Yinchenhao Tang (YCHT), whose efficacy in treating hepatic injury (HI) and Jaundice syndrome, has recently been well established as a case study. We also conducted a systematic analysis of synergistic effects of the principal compound using biochemistry, pharmacokinetics and systems biology, to explore the key molecular mechanisms. We had found that the three component (6,7-dimethylesculetin (D), geniposide (G), and rhein (R)) combination exerts a more robust synergistic effect than any one or two of the three individual compounds by hitting multiple targets. They can regulate molecular networks through activating both intrinsic and extrinsic pathways to synergistically cause intensified therapeutic effects. This paper provides an overview of the recent and potential developments of chemical fingerprinting coupled with systems biology advancing drug discovery towards more agile development of targeted combination therapies for the YCHT.
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Rapid discovery and global characterization of chemical constituents and rats metabolites of Phellodendri amurensis cortex by ultra-performance liquid chromatography-electrospray ionization/quadrupole-time-of-flight mass spectrometry coupled with pattern
Analyst
PUBLISHED: 04-24-2013
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To discover and screen the constituents or metabolites absorbed into blood after oral administration of herbal medicines tends to be more and more difficult. In this work, an integrative pattern recognition approach of principal component analysis (PCA) and orthogonal partial least squared discriminant analysis (OPLS-DA) was successfully applied for rapid discovery of natural compounds from herbal medicines. A rapid, sensitive, and reliable ultra performance liquid chromatography coupled with electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS) method with Masslynx™ software was established to characterize the chemical constituents and rats metabolites of Phellodendri amurensis cortex (Guan Huangbai, GHB). The analysis was performed on a Waters UPLC HSS T3 column (2.1 × 100 mm, 1.8 ?m) using gradient elution system. A hyphenated electrospray ionization and quadrupole-time-of-flight analyzer was used for the determination of accurate mass of the protonated or deprotonated molecule and fragment ion in both negative and positive modes. A total of 46 peaks were obtained, 41 of which were tentatively characterized from GHB. In the S-plot of OPLS-DA, 24 interested ions (17 ions in positive mode and 6 ions in negative mode) were extracted, among them, 12 absorbed prototype components of GHB and 12 metabolites were identified in vivo. Major metabolic reactions of GHB were demethylation, methylation and glucuronidation. This is the first report on systematic analysis of chemical constituents and in vivo metabolites of GHB. It is concluded that UPLC-MS coupled with pattern recognition approach for the identification of herbal constituents in biological samples has been successfully developed. The method can also be applied to rapid discovery and global characterization of the constituents in rat serum after oral administration of other herbal medicines.
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Profiling and identification of the absorbed constituents and metabolites of schisandra lignans by ultra-performance liquid chromatography coupled to mass spectrometry.
Biomed. Chromatogr.
PUBLISHED: 04-21-2013
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Schisandra chinensis Baill grows wild in Russia, China, Korea and Japan, and its fruit has been found to be effective in amnesia and insomnia. It is enriched in schisandra lignans (SL) that are major components responsible for therapeutic action. However, there are no reports on the biotransformation analysis of SL. An ultra-performance liquid chromatography/electrospray-ionization high-definition mass spectrometry (UPLC-Q-TOF-HDMS) method was developed to investigate the metabolism of SL in vivo. MS was performed on a Waters Micromass high-definition system with an electrospray ionization source in positive ion mode and automated MetaboLynx software analysis with excellent MS accuracy and enhanced MS data acquisition. An improved mass defect filter (MDF) method employing both drug and core structure filter templates was applied to the processing of UPLC-Q-TOF-HDMS data for the detection and structural characterization of metabolites. In this study, 30 metabolites were detected and identified in vivo, and demethylation and hydroxylation were confirmed as the primacy metabolic pathway for SL in rat plasma. In conclusion, the presently developed methodology was suitable for biotransformation research of SL and will find wide use in metabolic studies for other herbal medicines.
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Proteomics analysis of hepatoprotective effects for scoparone using MALDI-TOF/TOF mass spectrometry with bioinformatics.
OMICS
PUBLISHED: 03-20-2013
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Abstract Scoparone is an active ingredient of Yinchenhao (Artemisia annua L.), a well-known Chinese medicinal plant, and has been utilized in prevention and therapy of liver damage. However, the molecular drug targets associated with the pharmacological effects of scoparone are largely unknown. In the present article, we extend the previous research on Yinchenhao through a study of its active ingredient and thus the putative targets of scoparone. We employed two-dimensional gel electrophoresis, and all proteins expressed were identified by MALDI-TOF/TOF MS and database research. Protein-interacting networks and pathways were also mapped and evaluated. The possible protein network associated with scoparone was constructed, and contribution of these proteins to the protective effect of scoparone against the carbon tetrachloride-induced acute liver injury in rats are discussed herein. Hepatoprotective effects of scoparone on liver injury in rats were associated with regulated expression of six proteins which were closely related in our protein-protein interaction network, and appear to be involved in antioxidation and signal transduction, energy production, immunity, metabolism, and chaperoning. These observations collectively provide new insights on the molecular mechanisms of scoparone action against hepatic damage in rats.
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Recent advances in natural products from plants for treatment of liver diseases.
Eur J Med Chem
PUBLISHED: 03-04-2013
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Liver disease is any condition that may cause liver inflammation or tissue damage and affects liver function. Natural products that are found in vegetables, fruits, plant extracts, herbs, insects, and animals, have been traditionally used for treating liver diseases. They are chemical compounds that usually have biological activities for use in drug discovery and design. Many natural products have been clinically available as potent hepatoprotective agents against commonly occurring liver diseases. This review summarizes the current progress in the basic, clinical, and translational research on natural products in treatment of various liver diseases. Furthermore, we will focus on the discovery and biological evaluation of the natural products, which shows potential as a new therapeutic agent of liver diseases.
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Power of metabolomics in diagnosis and biomarker discovery of hepatocellular carcinoma.
Hepatology
PUBLISHED: 02-15-2013
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Hepatocellular carcinoma (HCC) is the commonest primary hepatic malignancy and the third most common cause of cancer-related death worldwide. Incidence remains highest in the developing world and is steadily increasing across the developed world. Current diagnostic modalities, of ultrasound and ?-fetoprotein, are expensive and lack sensitivity in tumor detection. Because of its asymptomatic nature, HCC is usually diagnosed at late and advanced stages, for which there are no effective therapies. Thus, biomarkers for early detection and molecular targets for treating HCC are urgently needed. Emerging high-throughput metabolomics technologies have been widely applied, aiming at the discovery of candidate biomarkers for cancer staging, prediction of recurrence and prognosis, and treatment selection. Metabolic profiles, which are affected by many physiological and pathological processes, may provide further insight into the metabolic consequences of this severe liver disease. Small-molecule metabolites have an important role in biological systems and represent attractive candidates to understand HCC phenotypes. The power of metabolomics allows an unparalleled opportunity to query the molecular mechanisms of HCC. This technique-driven review aims to demystify the metabolomics pathway, while also illustrating the potential of this technique, with recent examples of its application in HCC.
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Metabolomics coupled with proteomics advancing drug discovery toward more agile development of targeted combination therapies.
Mol. Cell Proteomics
PUBLISHED: 01-29-2013
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To enhance the therapeutic efficacy and reduce the adverse effects of traditional Chinese medicine, practitioners often prescribe combinations of plant species and/or minerals, called formulae. Unfortunately, the working mechanisms of most of these compounds are difficult to determine and thus remain unknown. In an attempt to address the benefits of formulae based on current biomedical approaches, we analyzed the components of Yinchenhao Tang, a classical formula that has been shown to be clinically effective for treating hepatic injury syndrome. The three principal components of Yinchenhao Tang are Artemisia annua L., Gardenia jasminoids Ellis, and Rheum Palmatum L., whose major active ingredients are 6,7-dimethylesculetin (D), geniposide (G), and rhein (R), respectively. To determine the mechanisms underlying the efficacy of this formula, we conducted a systematic analysis of the therapeutic effects of the DGR compound using immunohistochemistry, biochemistry, metabolomics, and proteomics. Here, we report that the DGR combination exerts a more robust therapeutic effect than any one or two of the three individual compounds by hitting multiple targets in a rat model of hepatic injury. Thus, DGR synergistically causes intensified dynamic changes in metabolic biomarkers, regulates molecular networks through target proteins, has a synergistic/additive effect, and activates both intrinsic and extrinsic pathways.
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Systems biology approach opens door to essence of acupuncture.
Complement Ther Med
PUBLISHED: 01-21-2013
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For World Health Organization proposed Health for All, we should be to promote the use of traditional medicine. Traditional Chinese acupuncture has a history over 3000 years and is effective in the treatment of many conditions with few side effects. Acupuncture, an intrinsic part of traditional Chinese medicine (TCM), which utilizes fine needles to pierce through specific anatomical points (called "acupoints"), has been extensively used and has emerged as an important modality of complementary and alternative therapy to Western medicine. It is vital and necessary to explore the underlying biological mechanisms of acupuncture. Systems biology has become practically available and resembles acupuncture in many aspects and is current key technology that serves as the major driving force for translation of acupuncture medicine revolution into practice, will advance acupuncture therapy into healthcare for individuals. High-throughput genomics, proteomics and metabolomics in the context of systems biology have been able to identify potential candidates for the effects of acupuncture and provide valuable information toward understanding mechanisms of the therapy. To realize the full potential of TCM acupuncture, we describe the current status of principles and practice of acupuncture integrated with systems biology platform in the post-genomic era. Some characteristic examples are presented to highlight the application of this platform in omics and systems biology approaches to acupuncture research and some of the necessary milestones for moving acupuncture into mainstream health care.
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Ultra-performance LC-ESI/quadrupole-TOF MS for rapid analysis of chemical constituents of Shaoyao-Gancao decoction.
J Sep Sci
PUBLISHED: 01-12-2013
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Shaoyao-Gancao decoction (SGD), a traditional Chinese formulae containing Paeoniae Radix and Glycyrrhizae Radix, is commonly used to relieve abdominal pain. It has attracted increasingly much attention as one of the most popular and valuable herbal medicine in clinic. However, the systematic analysis of chemical constituents of SGD are difficult to determine and thus remain unclear. In this paper, a rapid, sensitive, and reliable ultra-performance LC-ESI/quadrupole-TOF high-definition MS (UPLC-ESI-Q-TOF-MS) with automated MetaboLynx analysis in negative ion mode were established to characterize the chemical constituents of SGD. The analysis was performed on a Waters UPLC(TM) HSS T3 (2.1 × 100 mm, 1.8 ?m) using gradient elution system. MS/MS fragmentation behavior was proposed for aiding the structural identification of the components. With the optimized conditions, a total of 58 peaks were tentatively characterized by comparing the retention time and mass spectrometry data and retrieving the reference literatures. Of note, 44 ingredients were identified from Glycyrrhizae Radix, and 14 were from Paeoniae Radix. It is concluded that a rapid and robust platform based on UPLC-ESI-Q-TOF-MS was successfully developed for globally identifying multiple-constituent of traditional Chinese medicine prescriptions. This is the first report on systematic analysis of chemical constituents and in vivo metabolites of SGD.
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Pharmacokinetic study of schisandrin, schisandrol B, schisantherin A, deoxyschisandrin, and schisandrin B in rat plasma after oral administration of Shengmaisan formula by UPLC-MS.
J Sep Sci
PUBLISHED: 01-10-2013
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Shengmaisan (SMS) is a traditional Chinese medicine prescription widely used for the treatment of cardiovascular diseases in Asia. Its lignans are major components responsible for therapeutic action. A rapid and specific UPLC-Q-TOF/MS has been developed and validated for simultaneous quantification of the five main bioactive components, i.e. schisandrin, schisandrol B, schisantherin A, deoxyschisandrin, and schisandrin B, in rat plasma after oral administration of SMS. All calibration curves showed excellent linearity within the test ranges. Validation proved the repeatability of the method was good and recovery was satisfactory. The separation of these compounds was carried out on a Waters ACQUITY HSS T(3) column (2.1 × 100 mm, 1.8 ?m) by linear gradient elution using a mobile phase consisting of 0.01% formic acid in water and ACN containing 0.01% formic acid. In this work, plasma pharmacokinetic characteristics of lignans components after oral administration SMS were investigated using UPLC-Q-TOF/MS method. MS was performed on a Waters Micromass high-definition technology with an ESI source. Data were analyzed and estimated by compartmental methods and pharmacokinetic parameters calculated using WinNonlin Professional version 6.1. Results demonstrated that the proposed UPLC-Q-TOF/MS method was successfully applied to pharmacokinetic study of all components in rat plasma after oral administration of the SMS.
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Urinary metabolic profiling identifies a key role for glycocholic acid in human liver cancer by ultra-performance liquid-chromatography coupled with high-definition mass spectrometry.
Clin. Chim. Acta
PUBLISHED: 01-08-2013
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Metabolomics has been proposed to be a hallmark of cancer, yet a systematic characterization of a metabolite and metabolic pathways in human hepatocarcinoma (HCC) remains a challenge.
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Involvement of early growth response-2 (Egr-2) in lipopolysaccharide-induced neuroinflammation.
J. Mol. Histol.
PUBLISHED: 01-02-2013
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Early growth response-2 (Egr-2) protein is a transcription factor, which belongs to Egr family which involve in modulating the peripheral immune response, by means of the induction of differentiation of lymphocyte precursors, activation of T and B cells. Egr-2 plays essential roles in peripheral nerve myelination, adipogenesis, tissue repair and fibrosis, immune tolerance; however, its regulation and role in central nervous system (CNS) remain poorly understood. In contrast to Egr-1, which has been extensively investigated, the regulation and function of Egr-2 remains less well characterized. To elaborate whether Egr-2 was involved in CNS injury, we performed a neuroinflammatory model by lipopolysaccharide (LPS) lateral ventral injection in adult rats. Egr-2 expression was strongly induced in active glia cells (astrocytes and microglias) in inflamed brain cortex. In vitro studies indicated that the upregulation of Egr-2 may be involved in the subsequent glia cellular activation following LPS exposure; and knock down of Egr-2 in primary mixed glial cultures (MGC) by siRNA showed that Egr-2 promoted the synthesis of TNF-?. Collectively, these results suggested Egr-2 may be important in host defense in CNS immune response, which might provide a potential target to the treatment of neuroinflammation.
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Metabolomics and proteomics annotate therapeutic properties of geniposide: targeting and regulating multiple perturbed pathways.
PLoS ONE
PUBLISHED: 01-01-2013
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Geniposide is an important constituent of Gardenia jasminoides Ellis, a famous Chinese medicinal plant, and has displayed bright prospects in prevention and therapy of hepatic injury (HI). Unfortunately, the working mechanisms of this compound are difficult to determine and thus remain unknown. To determine the mechanisms that underlie this compound, we conducted a systematic analysis of the therapeutic effects of geniposide using biochemistry, metabolomics and proteomics. Geniposide significantly intensified the therapeutic efficacy as indicated by our modern biochemical analysis. Metabolomics results indicate 9 ions in the positive mode as differentiating metabolites which were associated with perturbations in primary bile acid biosynthesis, butanoate metabolism, citrate cycle (TCA cycle), alanine, aspartate and glutamate metabolism. Of note, geniposide has potential pharmacological effect through regulating multiple perturbed pathways to normal state. In an attempt to address the benefits of geniposide based on the proteomics approaches, the protein-interacting networks were constructed to aid identifying the drug targets of geniposide. Six identified differential proteins appear to be involved in antioxidation and signal transduction, energy production, immunity, metabolism, chaperoning. These proteins were closely related in the protein-protein interaction network and the modulation of multiple vital physiological pathways. These data will help to understand the molecular therapeutic mechanisms of geniposide on hepatic damage rats. We also conclude that metabolomics and proteomics are powerful and versatile tools for both biomarker discovery and exploring the complex relationships between biological pathways and drug response, highlighting insights into drug discovery.
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Urinary metabolic biomarker and pathway study of hepatitis B virus infected patients based on UPLC-MS system.
PLoS ONE
PUBLISHED: 01-01-2013
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Hepatitis B virus (HBV) is the fatal consequence of chronic hepatitis, and lack of biomarkers has been a long standing bottleneck in the clinical diagnosis. Metabolomics concerns with comprehensive analysis of small molecules and provides a powerful approach to discover biomarkers in biological systems. Here, we present metabolomics analysis applying ultra-performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry. (UPLC-Q-TOF-HDMS) to determine metabolite alterations in HBV patients. Most important permutations are elaborated using multivariate statistical analysis and network analysis that was used to select the metabolites for the noninvasive diagnosis of HBV. In this study, the total 11 urinary differential metabolites were identified and contributed to HBV progress involving several key metabolic pathways by using pathway analysis with MetPA, which are promising biomarker candidates for diagnostic research. More importantly, of 11 altered metabolites, 4 metabolite markers were effective for the diagnosis of human HBV, achieved a satisfactory accuracy, sensitivity and specificity, respectively. It demonstrates that metabolomics has the potential as a non-invasive tool to evaluate the potential of these metabolites in the early diagnosis of HBV patients. These findings may be promising to yield a valuable insight into the pathophysiology of HBV and to advance the approaches of diagnosis, treatment, and prevention.
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Pattern recognition approaches and computational systems tools for ultra performance liquid chromatography-mass spectrometry-based comprehensive metabolomic profiling and pathways analysis of biological data sets.
Anal. Chem.
PUBLISHED: 12-19-2011
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Metabolomics represents an emerging and powerful discipline that provides an accurate and dynamic picture of the phenotype of biosystems through the study of potential metabolites that could be used for therapeutic targets and discovery of new drugs. Metabolomic network construction has led to the integration of metabolites associated with the caused perturbation of multiple pathways. Herein, we present a method for the construction of efficient networks with regard to that Jujuboside B (JuB) protects against insomnia as a case study. UPLC/ESI-SYNAPT-HDMS coupled with pattern recognition methods including PCA, PLS-DA, OPLS-DA, and computational systems analysis were integrated to obtain comprehensive metabolomic profiling and pathways of the large biological data sets. Among the regulated pathways, twelve biomarkers were identified and tryptophan metabolism, phenylalanine, tyrosine, tryptophan biosynthesis, arachidonic acid metabolism, and phenylalanine metabolism related network were acutely perturbed. Results not only supplied a systematic view of the development and progression of insomnia but also were used to analyze the therapeutic effects of JuB, a widely used anti-insomina medicine in clinics. The results showed that JuB administration could provide satisfactory effects on insomnia through partially regulating the perturbed pathway. We have constructed a metabolomic feature network of JuB to protect against insomnia. The most promising use in the near future would be to clarify pathways for the drugs and get biomarkers for these pathways, to help guide testable predictions, provide insights into drug action mechanisms, and enable us to increase research productivity toward metabolomic drug discovery.
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Metabolomics study on the toxicity of aconite root and its processed products using ultraperformance liquid-chromatography/electrospray-ionization synapt high-definition mass spectrometry coupled with pattern recognition approach and ingenuity pathways an
J. Proteome Res.
PUBLISHED: 12-02-2011
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The mother and lateral root of Aconitum carmichaelii Debx, named "Chuanwu" (CW) and "Fuzi", respectively, has been used to relieve joint pain and treat rheumatic diseases for over 2000 years. However, it has a very narrow therapeutic range, and the toxicological risk of its usage remains very high. The traditional Chinese processing approach, Paozhi (detoxifying measure),can decompose poisonous Aconitum alkaloids into less or nontoxic derivatives and plays an important role in detoxification. The difference in metabolomic characters among the crude and processed preparations is still unclear, limited by the lack of sensitive and reliable biomarkers. Therefore, this paper was designed to investigate comprehensive metabolomic characters of the crude and its processed products by UPLC-Q-TOF-HDMS combined with pattern recognition methods and ingenuity pathway analysis (IPA). The significant difference in metabolic profiles and changes of metabolite biomarkers of interest between the crude and processed preparations were well observed. The underlying regulations of Paozhi-perturbed metabolic pathways are discussed according to the identified metabolites, and four metabolic pathways are identified using IPA. The present study demonstrates that metabolomic analysis could greatly facilitate and provide useful information to further comprehensively understand the pharmacological activity and potential toxicity of processed Aconite roots in the clinic.
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[Insertion of glycoprotein gene between P and M gene influences the pathogenicity of the rabies virus Flury LEP].
Wei Sheng Wu Xue Bao
PUBLISHED: 11-22-2011
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To study the biological characteristics and pathogenicity of a recombinant rabies virus Flury LEP (low egg passage) that has two glycoprotein genes (G gene).
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Modern analytical techniques in metabolomics analysis.
Analyst
PUBLISHED: 11-21-2011
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Metabolomics is the comprehensive assessment of endogenous metabolites and attempts to systematically identify and quantify metabolites from a biological sample. Small-molecule metabolites have an important role in biological systems and represent attractive candidates to understand disease phenotypes. Metabolites represent a diverse group of low-molecular-weight structures including lipids, amino acids, peptides, nucleic acids, organic acids, vitamins, thiols and carbohydrates, which makes global analysis a difficult challenge. The recent rapid development of a range of analytical platforms, including GC, HPLC, UPLC, CE coupled to MS and NMR spectroscopy, could enable separation, detection, characterization and quantification of such metabolites and related metabolic pathways. Owing to the complexity of the metabolome and the diverse properties of metabolites, no single analytical platform can be applied to detect all metabolites in a biological sample. The combined use of modern instrumental analytical approaches has unravelled the ideal outcomes in metabolomics, and is beneficial to increase the coverage of detected metabolites that can not be achieved by single-analysis techniques. Integrated platforms have been frequently used to provide sensitive and reliable detection of thousands of metabolites in a biofluid sample. Continued development of these analytical platforms will accelerate widespread use and integration of metabolomics into systems biology. Here, the application of each hyphenated technique is discussed and its strengths and limitations are discussed with selected illustrative examples; furthermore, this review comprehensively highlights the role of integrated tools in metabolomic research.
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[Dao-di herbs and its change of cultivated origin place].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 10-29-2011
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The origin place change is an almost universal rule for many Chinese medicines according to the formation and change history of ancient and recent Dao-di herbs. Among them, the quality and output are the essence and driving force of the formation and change of Dao-di herbs origin place. More and more species and quantities of Chinese medicine were cultivated along with development of Chinese medicine and modern manufacturing technology, and its production regions far exceed natural location zones, the rationality of origin place change should be based on drug quality evaluation. Under the guidance of the formation mode of Dao-di herbs, characteristics of Dao-di herbs were summarized, its characteristics and applied limits were suggested, some traditional Chinese medicinal materials only with higher output, cultivated materials in distribution area for wild, or materials with poor genetic resources failing to answer with Dao-di herbs. Chinese medicines authenticity should strengthen the study of germplasm and drug quality assessment, the suitability of producing area should be assessed according to output and quality under the guidance of suitable locations regionalization.
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Metabolomics study on Fuzi and its processed products using ultra-performance liquid-chromatography/electrospray-ionization synapt high-definition mass spectrometry coupled with pattern recognition analysis.
Analyst
PUBLISHED: 10-26-2011
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The lateral root of Aconitum carmichaelii Debx is named "Fuzi" which is widely distributed across Asia and North America and has been used to relieve joint pain and treat rheumatic diseases for over two thousand years. However, it has very narrow therapeutic ranges and despite the toxicological risk, its usage remains very high. A traditional Chinese processing approach (Paozhi, detoxifying measure) is necessary to remove the poisonous Aconitum alkaloids mainly deriving from the diester diterpene alkaloids (DDAs) including aconitine, mesaconitine and hypaconitine. They can be decomposed into less or non-toxic derivatives through Paozhi that plays an essential role in detoxification. Processed Fuzi is mainly focused on the three main forms of Yanfuzi (YFZ), Heishunpian (HSP) and Baifupian (BFP) which are highly desirable in order to guarantee the clinical safety and their low toxicity in decoctions. The difference in metabolomic characters between Fuzi and its processed preparations is still completely unclear. Therefore, this paper was designed to investigate a comprehensive metabolome of Fuzi and its processed products by ultra-performance liquid-chromatography/electrospray-ionization synapt high-definition mass spectrometry (UPLC-Q-TOF-HDMS) combined with pattern recognition methods. The difference in metabolic profiles between Fuzi and its processed preparations was well observed by the principal component analysis (PCA) of the MS spectra. Significant changes of 19 metabolite biomarkers were detected in the Fuzi samples and three preparations. The underlying regulations of Paozhi-perturbed metabolic pathways were also discussed according to the identified metabolites. The present study proves that UPLC-Q-TOF-HDMS based metabolomic analysis greatly contributes to the investigation of Fuzi metabolism through Paozhi techniques, and provides useful information to further comprehensively understand the pharmacological activity and potential toxicity of processed Fuzi in a clinical environment.
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Pharmacokinetics screening for multi-components absorbed in the rat plasma after oral administration traditional Chinese medicine formula Yin-Chen-Hao-Tang by ultra performance liquid chromatography-electrospray ionization/quadrupole-time-of-flight mass s
Analyst
PUBLISHED: 10-12-2011
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Traditional Chinese medicine (TCM) has been widely used in many oriental countries for thousands of years and played an indispensable role in the prevention and treatment of diseases, especially the complicated and chronic ones. It is a very complex mixture containing hundreds or thousands of different components. Pharmacokinetic study on active constituents in TCM preparations is a good way for us to explain and predict a variety of events related to the efficacy and toxicity of TCM. A selective and sensitive method of ultra performance liquid chromatography coupled with electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS/MS) was first developed to screen the potentially bioactive components in vivo, using the semi-quantitative determination of multicomponents in the rat plasma after a single oral administration of Yin-Chen-Hao-Tang (YCHT), a famous TCM formula for liver disorders. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) were built to evaluate the differences of pharmacokinetic behaviors (time-course) of the absorbed components of YCHT. Here, we report that the developed method was successfully applied to monitoring the pharmacokinetic time-course of 21 compounds in rat plasma, and were grouped in 3 separate clusters using pattern recognition approaches (both HCA and PCA). Comparing the body dynamics of each composition, the initial choice of the following 9 compounds as the candidate components was: 7-methoxycoumarin-6-hydroxyl sulfate, genipingentiobioside, geniposide, 6,7-dimethylesculetin, peak 16, chimaphylin, 6-dementhoxycapillarisin, capillarisin, rhein. Pharmacokinetics based-UPLC-ESI-Q-TOF-MS/MS combined with HCA and PCA approaches can provide a reliable and suitable means of identifying and screening potentially bioactive components contributing to pharmacological effects of TCM, further prospecting natural products in the search for new leads in drug discovery.
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Potential drug targets on insomnia and intervention effects of Jujuboside A through metabolic pathway analysis as revealed by UPLC/ESI-SYNAPT-HDMS coupled with pattern recognition approach.
J Proteomics
PUBLISHED: 10-10-2011
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Potential metabolites from the metabolic pathways could be therapeutic targets and useful for the discovery of broad spectrum drugs. UPLC/ESI-SYNAPT-HDMS coupled with pattern recognition methods including PCA, PLS-DA, OPLS-DA and Heatmap were integrated to examine the global metabolic signature of insomnia and intervention effects of Jujuboside A (JuA). Six unique pathways of the insomnia were identified using Ingenuity Pathway Analysis (IPA) software. The VIP-value threshold cutoff of the metabolites was set to 10, above this threshold, were filtered out as potential target biomarkers. Sixteen distinct metabolites were identified from these pathways, and 6 of them can be considered for rational drug design. It was further experimental validation that the changes in metabolic profiling were restored to their baseline values after JuA treatment according to the multivariate data analysis. Potential metabolite network of the insomnia was preliminarily predicted JuA-target interaction networks, and could be further explored for in silico docking studies with suitable drugs. Thus, our method is an efficient procedure for drug target identification through metabolic analysis. It can guide testable predictions, provide insights into drug action mechanisms and enable us to increase research productivity toward metabolomic drug discovery.
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Recent and potential developments of biofluid analyses in metabolomics.
J Proteomics
PUBLISHED: 10-04-2011
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Metabolomics, one of the omic sciences in systems biology, is the global assessment and validation of endogenous small-molecule metabolites within a biologic system. Analysis of these key metabolites in body fluids has become an important role to monitor the state of biological organisms and is a widely used diagnostic tool for disease. A majority of these metabolites are being applied to metabolic profiling of the biological samples, for example, plasma and whole blood, serum, urine, saliva, cerebrospinal fluid, synovial fluid, semen, and tissue homogenates. However, the recognition of the need for a holistic approach to metabolism led to the application of metabolomics to biological fluids for disease diagnostics. A recent surge in metabolomic applications which are probably more accurate than routine clinical practice, dedicated to characterizing the biological fluids. While developments in the analysis of biofluid samples encompassing an important impediment, it must be emphasized that these biofluids are complementary. Metabolomics provides potential advantages that classical diagnostic approaches do not, based on following discovery of a suite of clinically relevant biomarkers that are simultaneously affected by the disease. Emerging as a promising biofocus, metabolomics will drive biofluid analyses and offer great benefits for public health in the long-term.
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Metabolomic study of insomnia and intervention effects of Suanzaoren decoction using ultra-performance liquid-chromatography/electrospray-ionization synapt high-definition mass spectrometry.
J Pharm Biomed Anal
PUBLISHED: 08-29-2011
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Metabolomics is the comprehensive assessment of endogenous metabolites of a biological system in a holistic context, and its property consists with the global view of traditional Chinese medicine (TCM). Suanzaoren decoction (SZRD), an ancient TCM formulae, has been used for treating insomnia for centuries, and its mechanism remains unclear completely. This paper was designed to explore globally metabolomic characters of the insomnia and the therapeutic effects of SZRD. Ultra-performance liquid-chromatography/electrospray-ionization synapt high-definition mass spectrometry (UPLC/ESI-SYNAPT-HDMS) combined with pattern recognition approaches including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA) and orthogonal projection to latent structures discriminate analysis (OPLS-DA) were integrated to approximate the comprehensive metabolic signature and discover differentiating metabolites. The changes in metabolic profiling were restored to their baseline values after SZRD treatment according to the PCA score plots. Altogether, the current metabonomic approach based on UPLC/ESI-SYNAPT-HDMS indicate 20 ions (9 in the negative mode, 11 in the positive mode) as "differentiating metabolites". The alterations in these metabolites were associated with perturbations in amino acid and fatty acid metabolism, in response to insomnia through immune and nervous system. Of note, we found that SZRD increases sleep activity and exhibits binding affinity for serotonin receptors. These results implicate the therapeutic effects of SZRD may mediate through serotonergic activation. Our findings also show the robust UPLC/ESI-SYNAPT-HDMS techniques is promising for metabolites profiling analysis of TCM and open new perspectives to using metabolomics platform to resolve special TCM issues.
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Ultra-performance liquid chromatography coupled with electrospray ionization/quadrupole-time-of-flight mass spectrometry for rapid analysis of constituents of Suanzaoren decoction.
J Sep Sci
PUBLISHED: 07-18-2011
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A rapid, sensitive, specific and reliable ultra-performance liquid chromatography coupled with electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS) method with MassLynx™ MassFragment was developed for the analysis of Suanzaoren decoction (SZRD), a Chinese herbal prescription. The analysis was performed on a Waters UPLC BEH C(18) column using gradient elution system. A hyphenated electrospray ionization and Q-TOF analyzer was used for the determination of accurate mass of the protonated or deprotonated molecule and fragment ion in both negative and positive modes. The chromatographic separation was achieved by UPLC, which used a column with 1.7 ?m particle packing which enabled higher speed of analysis, peak capacity, greater resolution and increased sensitivity. The constituents of SZRD were identied and confirmed according to the mass spectrometric fragmentation mechanisms, MS/MS fragment ions, relevant literature and the establishment of an in-house molecular formula database. With this method, a total of 22 compounds of SZTD were tentatively identied based on MS and MS/MS data and comparison with available databases. It is concluded that a rapid and robust platform based on UPLC-ESI-Q-TOF-MS was established, which is useful for identifying multiple-constituent of traditional Chinese medicine (TCM) prescriptions. Our present results proved that the established method could provide helpful chemical information for further pharmacological mechanism research of SZRD.
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Simultaneous in vivo RP-HPLC-DAD quantification of multiple-component and drug-drug interaction by pharmacokinetics, using 6,7-dimethylesculetin, geniposide and rhein as examples.
Biomed. Chromatogr.
PUBLISHED: 07-03-2011
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Increasing evidence has demonstrated that multidrug combinations could amplify the therapeutic efficacies of each agent. Interestingly, the pharmacological effect of traditional Chinese medicine (TCM) is usually attributed to the drug-interaction property (synergism) of multiple active constituents. Pharmacokinetics is a useful means of evaluating the drug interactions of major active compounds in TCM. A simple, sensitive and reliable RP-HPLC-DAD method has been developed to simultaneously quantify 6,7-dimethylesculetin (D), geniposide (G) and rhein (R), which are the active ingredients in Yin-Chen-Hao-Tang (YCHT), performing drug-interaction pharmacokinetics studies in vivo. Plasma samples were prepared using methanolic precipitation, a filtration step, and then injection of the methanolic extract onto a Nova-Pak C?? Guard-Pak™ guard column with a gradient mobile phase. Triple-wavelength diode array detection was set at ?(max) values of 343?nm for D, 241?nm for the G, and 259?nm for R. Our results successfully demonstrate that this method has excellent and satisfactory selectivity, sensitivity, linearity, precision, accuracy and recovery. In healthy rats, the estimated pharmacokinetic parameters (i.e. C(max) , AUC and Cl) of D, G and R, when administered with COC (a combination of D, G and R), were C(max) 16.05?mg/L, AUC 108.96?mg?h/L and Cl 0.36?L/h for D; C(max) 9.35?mg/L, AUC 64.71?mg?h/L and Cl 0.88?L/h for G; and C(max) 14.18?mg/L, AUC 57.98?mg?h/L and Cl 1.77?L/h for R. Here, we report that the COC combination could significantly increase the plasma level and slow the elimination rate compared with any one or two of the three individual compounds, which may indicate a drug-drug interaction.
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An in vivo analysis of the therapeutic and synergistic properties of Chinese medicinal formula Yin-Chen-Hao-Tang based on its active constituents.
Fitoterapia
PUBLISHED: 06-27-2011
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6,7-Dimethylesculetin (D), geniposide (G) and rhein (R) are the three major active ingredients of Yin-Chen-Hao-Tang (YCHT), a famous Chinese herbal formula, which has been shown to be clinically effective for treating hepatic injury (HI) syndrome. The present study was conducted to investigate the therapeutic and synergistic effects of COC (combination of D, G and R) on HI rats by combining pharmacokinetic with biochemical analysis strategy. Plasma was analyzed by using reversed-phase high performance liquid chromatography (RP-HPLC). Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) models were built to evaluate the therapeutic and synergistic effects of COC at the biochemical level. Here, we report that the COC combination could increase the plasma level, slow elimination rate, exert a more robust therapeutic effect than any one or two of the three individual compounds by hitting multiple targets in a rat model of HI. Overall, this beneficially accounts for the popular view that traditional Chinese medicine (TCM) formula usually takes multi-component to exert their therapeutic effects. We suggest that dissecting the mode of action of clinically effective formula to be capable of producing a sufficient effect at low doses.
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Newcastle disease virus-vectored rabies vaccine is safe, highly immunogenic, and provides long-lasting protection in dogs and cats.
J. Virol.
PUBLISHED: 06-01-2011
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Effective, safe, and affordable rabies vaccines are still being sought. Newcastle disease virus (NDV), an avian paramyxovirus, has shown promise as a vaccine vector for mammals. Here, we generated a recombinant avirulent NDV La Sota strain expressing the rabies virus glycoprotein (RVG) and evaluated its potential to serve as a vaccine against rabies. The recombinant virus, rL-RVG, retained its high-growth property in chicken eggs, with titers of up to 10?·? 50% egg infective doses (EID??)/ml of allantoic fluid. RVG expression enabled rL-RVG to spread from cell to cell in a rabies virus-like manner, and RVG was incorporated on the surface of the rL-RVG viral particle. RVG incorporation did not alter the trypsin-dependent infectivity of the NDV vector in mammalian cells. rL-RVG and La Sota NDV showed similar levels of sensitivity to a neutralization antibody against NDV and similar levels of resistance to a neutralization antibody against rabies virus. Animal studies demonstrated that rL-RVG is safe in several species, including cats and dogs, when administered as multiple high doses of recombinant vaccine. Intramuscular vaccination with rL-RVG induced a substantial rabies virus neutralization antibody response and provided complete protection from challenge with circulating rabies virus strains. Most importantly, rL-RVG induced strong and long-lasting protective neutralization antibody responses to rabies virus in dogs and cats. A low vaccine dose of 10?·³ EID?? completely protected dogs from challenge with a circulating strain of rabies virus for more than a year. This is the first study to demonstrate that immunization with an NDV-vectored vaccine can induce long-lasting, systemic protective immunity against rabies.
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Future perspectives of personalized medicine in traditional Chinese medicine: a systems biology approach.
Complement Ther Med
PUBLISHED: 05-24-2011
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Deconstruction of molecular pathways and advances in enabling technology platforms have opened new horizons for disease management, exploring therapeutic solutions to each individual patient beyond the one-size fits all practice. Application of personalized medicine paradigms aims to achieve the right diagnosis and right treatment for the right patient at the right time at the right cost. With the potential to transform medical practice across global communities, personalized medicine is emerging as the flagship of modern medicine. In recent years, the health care paradigm has shifted from a focus on diseases to a major hot of personalized traditional Chinese medicine (TCM) with holistic approach. TCM focuses on health maintenance, emphasizes on enhancing the bodys resistance to diseases and especially showes great advantages in early intervention, personalized and combination therapies, etc. Systems biology, a new science of the 21st century, becomes practically available and resembles TCM in many aspects such as study method and design, and is current key component technologies that serves as the major driving force for translation of the personalized medicine revolution of TCM principles into practice, will advance personalized therapy principles into healthcare management tools for individuals and populations. Such system approach concepts are transforming principles of TCM to modern therapeutic approaches, enable a predictive and preventive medicine and will lead to personalized medicine. To realise the full potential of personalized TCM, we describe the current status of principles and practice of TCM integrated with systems biology platform. Some characteristic examples are presented to highlight the application of this platform to personalized TCM research and development as well as some of the necessary milestones for moving TCM into mainstream health care.
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Generation of a recombinant rabies Flury LEP virus carrying an additional G gene creates an improved seed virus for inactivated vaccine production.
Virol. J.
PUBLISHED: 05-16-2011
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The rabies Flury Low Egg Passage virus (LEP) has been widely used as a seed virus to generate inactive vaccine. Here, we established a reverse genetic system for LEP and generated a recombinant LEP virus (rLEP-G) that carries two identical G genes. This recombinant virus showed similar properties to those of LEP with respect to in vitro growth, neurotropism index, and virulence in mice. rLEP-G produced 4.3-fold more G protein than did LEP in BHK-21 cells. The inactivated vaccine generated from rLEP-G induced significantly higher virus neutralization titers in mice and dogs than those produced in response to LEP-derived vaccine. Our results suggest that rLEP-G is an improved seed virus candidate for inactivated rabies virus vaccine manufacture.
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Ultra-performance liquid chromatography coupled to mass spectrometry as a sensitive and powerful technology for metabolomic studies.
J Sep Sci
PUBLISHED: 04-13-2011
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Metabolomics is the comprehensive assessment of endogenous metabolites of a biological system. These large-scale analyses of metabolites are intimately bound to advancements in ultra-performance liquid chromatography-electrospray (UPLC) technologies and have emerged in parallel with the development of novel mass analyzers and hyphenated techniques. Recently, the combination of UPLC with MS covers a number of polar metabolites, thus enlarging the number of detected analytes in the widely used separation sciences. This technology has rapidly been accepted by the analytical community and is being gradually applied to various fields such as metabolomics and traditional Chinese medicine (TCM). Given the power of the technology, metabolomics has become increasingly popular in drug development, molecular medicine, traditional medicine and other biotechnology fields, since it profiles directly the phenotype and changes thereof in contrast to other "-omics" technologies. Hyphenated UPLC/MS technique is becoming a useful tool in the study of body fluids, represents a promising hyphenated microseparation platform in metabolomics and has a strong potential to contribute to disease diagnosis. This review describes the applications of UPLC/MS in metabolomic research, and comparison role of HPLC/MS, NMR and GC/MS, highlights its advantages and limitations with certain characteristic examples in the life and TCM sciences.
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Establishment of a stable CHO cell line with high level expression of recombinant porcine IFN-?.
Cytokine
PUBLISHED: 04-01-2011
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A CHO cell clone (CHO-PoIFN-?) with stable porcine IFN-? expression under control of CMV promoter was selected under G418 pressure. In a 25cm(2) cell culture flask (5 ml culture medium), the cumulative protein yield of recombinant PoIFN-? reached 2.3×10(6) IU/ml. This cells clone maintained stable expression for at least 20 generations even in the absence of G418 selection pressure. The expressed recombinant PoIFN-? could induce the expression of porcine Mx protein in PK15 cells, and activate the chicken Mx promoter-controlled luciferase reporter gene expression, confirming that the recombinant PoIFN-? has the biological activity of natural porcine type-I interferon. In addition, the recombinant PoIFN-? fully protected PK15 cells against 1000 TCID(50) of porcine transmissible gastroenteritis virus and pseudo-rabies virus infection, demonstrating its high potential in therapeutic applications. This is the first report of establishing a mammalian cell line with stable expression of porcine IFN-?.
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Rapid and global detection and characterization of the constituents in ShengMai San by ultra-performance liquid chromatography-high-definition mass spectrometry.
J Sep Sci
PUBLISHED: 03-20-2011
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An ultra-performance liquid chromatography-high definition mass spectrometry (UPLC-HDMS) method was developed for detection and characterization of the chemical constituents in ShengMai San (SMS), a traditional Chinese medical formula (TCMF). The full-scan LC-MS/MS data sets combined with extra mass were acquired within 14 min using UPLC-HDMS in the MS(E) mode in a single injection. As a result, 92 compounds were identified by comparing the accurate mass and fragments information with that of the authentic standards as well as by MS analysis and the correlative references data. These constituents included ginsenosides, lignans, steroidal saponins and homoisoflavanones. Among them, 25-hydroxyginsenosides were discovered in SMS for the first time. Compare with the previous studies, our research detected more compounds and presented more rapid by applying UPLC-HDMS. It is concluded that a rapid and effective method has been established based on UPLC-HDMS with the utilization of MS(E) , which shows high sensitivity and resolution that is suitable for identifying the constituents of SMS, and this method could be applied to other TCMF.
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Potential role of metabolomics apporoaches in the area of traditional Chinese medicine: as pillars of the bridge between Chinese and Western medicine.
J Pharm Biomed Anal
PUBLISHED: 01-27-2011
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Traditional Chinese medicine (TCM) is a holistic approach to health that attempts to bring the body, mind and spirit into harmony. Entering 21st century, TCM is getting more and more popular in the whole world for improving health condition of human beings and preventing or healing diseases, especially shows great advantages in early intervention, combination therapies and personalized medicine, etc. However, like almost all other ethnopharmacology, TCM also faces severe challenges and suffers from insufficient modern research owing to lack of scientific and technologic approaches, restricts the development of TCM in the world. Fortunately, a novel analytical technique, metabolomics (or metabonomics), adopts a top-down strategy to reflect the function of organisms from terminal symptoms of metabolic network and understand metabolic changes of a complete system caused by interventions in holistic context. Its property consists with the holistic thinking of TCM, may beneficially provide an opportunity to scientifically express the meaning of evidence-based Chinese medicine, such as Chinese medicine syndromes (CMS), preventive treatment, action of Chinese medicine, Chinese medical formulae (CMF) and acupuncture efficacy. This review summarizes potential applications of robust metabolomics apporoaches in the area of traditional oriental medicine, and highlights the key role of metabolomics to resolve special TCM issues.
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Metabolomic study of a rat fever model induced with 2,4-dinitrophenol and the therapeutic effects of a crude drug derived from Coptis chinensis.
Am. J. Chin. Med.
PUBLISHED: 01-08-2011
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This study describes the metabonomics of fevers in animal models and the therapeutic effects of Rhizoma coptidis extract (RCE) on them. The rat urinary samples were analyzed by UPLC/ ESI-Q-TOF/MS, combined with principal component analysis (PCA). Nine ions were chosen to characterize the similarities and differences in the responses to fever. The ion at m/z 206.0278 was unambiguously identified to be xanthurenic acid. This study demonstrated that the metabonomic approach can readily distinguish between febrile and healthy individuals. This data support the contention that the metabonomic approach represents a promising new technology for the development of rapid-throughput in vivo fever screening. Furthermore, this approach can detect the interfering effects of RCE. This investigation has led the authors to believe that metabonomics is a valid approach for explaining the therapeutic effects of traditional Chinese medicine on fevers.
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Metabolomics: towards understanding traditional Chinese medicine.
Planta Med.
PUBLISHED: 11-05-2010
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Metabolomics represent a global understanding of metabolite complement of integrated living systems and dynamic responses to the changes of both endogenous and exogenous factors and has many potential applications and advantages for the research of complex systems. As a systemic approach, metabolomics adopts a "top-down" strategy to reflect the function of organisms from the end products of the metabolic network and to understand metabolic changes of a complete system caused by interventions in a holistic context. This property agrees with the holistic thinking of Traditional Chinese Medicine (TCM), a complex medical science, suggesting that metabolomics has the potential to impact our understanding of the theory behind the evidence-based Chinese medicine. Consequently, the development of robust metabolomic platforms will greatly facilitate, for example, the understanding of the action mechanisms of TCM formulae and the analysis of Chinese herbal (CHM) and mineral medicine, acupuncture, and Chinese medicine syndromes. This review summarizes some of the applications of metabolomics in special TCM issues with an emphasis on metabolic biomarker discovery.
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The pharmacological effects of morroniside and loganin isolated from Liuweidihuang Wan, on MC3T3-E1 cells.
Molecules
PUBLISHED: 09-06-2010
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Liuweidihuang wan (LW), initially a well-known formula for curing "wu chi wu ruan", is commonly used nowadays for clinical treatment of postmenopausal osteoporosis (PO), but the identity of the effective substance(s) remains unclear. The present study was designed to evaluate the effects of morroniside and loganin isolated from LW on the proliferation, differentiation and apoptosis of MC3T3-E1 cells, as well as the possible mechanism of action. Morroniside and loganin had no effects on the proliferation of MC3T3-E1 cells, but both susbtances could improve the activity of alkaline phosphatase (ALP), and increase the contents of collagen type I and osteocalcin. Simultaneously, the mRNA expression of caspase-3, capase-9, RANKL was down-regulated and that of bcl-2 was up-regulated, which partially explains the anti-osteoporosis mechanism in MC3T3-E1 cells. In conclusion, morroniside and loganin may directly promote the differentiation and inhibit the apoptosis of MC3T3-E1 cells, and accordingly indirectly reduce bone resorption, which makes them promising natural drugs leads for treating PO in the near future.
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[Elucidation of compatibility principle and scientific value of Chinese medical formulae based on pharmacometabolomics].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 08-17-2010
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By using UPLC coupled with MS, the analysis of the constituents absorbed into blood after oral administration of Yinchenhao decotion with different combination had been carried out, the body fate of constituents detected in the blood was elucidated also; At the same time, the overall body respones after multi-input of the medical formule were evaluated by metabolomic analysis. These data explained the scientific values in both drug metabolisim and metabonomics, the results had given some new evidences for the potentialities of pharmacometabonomics to elucidate the compatibility principle of Chinese medical formule. As a conclusion, concept of "Pharmacometabolomics of Chinmediformulae" has been defined.
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Molecular basis of neurovirulence of flury rabies virus vaccine strains: importance of the polymerase and the glycoprotein R333Q mutation.
J. Virol.
PUBLISHED: 06-10-2010
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The molecular mechanisms associated with rabies virus (RV) virulence are not fully understood. In this study, the RV Flury low-egg-passage (LEP) and high-egg-passage (HEP) strains were used as models to explore the attenuation mechanism of RV. The results of our studies confirmed that the R333Q mutation in the glycoprotein (G(R333Q)) is crucial for the attenuation of Flury RV in mice. The R333Q mutation is stably maintained in the HEP genome background but not in the LEP genome background during replication in mouse brain tissue or cell culture. Further investigation using chimeric viruses revealed that the polymerase L gene determines the genetic stability of the G(R333Q) mutation during replication. Moreover, a recombinant RV containing the LEP G protein with the R333Q mutation and the HEP L gene showed significant attenuation, genetic stability, enhancement of apoptosis, and immunogenicity. These results indicate that attenuation of the RV Flury strain results from the coevolution of G and L elements and provide important information for the generation of safer and more effective modified live rabies vaccine.
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Rapid and global detection and characterization of aconitum alkaloids in Yin Chen Si Ni Tang, a traditional Chinese medical formula, by ultra performance liquid chromatography-high resolution mass spectrometry and automated data analysis.
J Pharm Biomed Anal
PUBLISHED: 03-04-2010
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An improved method employing Metabolynx XS with mass defect filter (MDF), a post-acquisition data processing software, was developed and applied for global detection of aconitum alkaloids in Yin Chen Si Ni Tang, a traditional Chinese medical formula (TCMF). The full-scan LC-MS/MS data sets with extra mass were acquired using ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) with the MS(E) mode in a single injection. To remove the interferences, Metabolynx XS was optimized to extract the ions of aconitum alkaloids located at the lower abundance. As a result, 62 ions were assigned rapidly to aconitum alkaloids and identified tentatively by comparing the accurate mass and fragments information with that of the authentic standards or by mass spectrometry analysis and retrieving the reference literatures. Compared with the previous studies on Fuzi-containing TCMF, the report detected more aconitum alkaloids, and the analysis process was accelerated by automated data processing. It is concluded that the screening capability of Metabolynx XS with MDF, together with the utilization of MS(E) in structural elucidation, can facilitate a rapid and comprehensive searching and effective structural characterization of aconitum alkaloids in TCMF.
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Thyroxine and reserpine-induced changes in metabolic profiles of rat urine and the therapeutic effect of Liu Wei Di Huang Wan detected by UPLC-HDMS.
J Pharm Biomed Anal
PUBLISHED: 02-26-2010
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The promise of metabonomics, a new "omics" technique, to validate Chinese medicines and the compatibility of Chinese formulas has been appreciated. The present study was undertaken to explore the excretion pattern of low molecular mass metabolites in the male Wistar-derived rat model of kidney yin deficiency induced with thyroxine and reserpine as well as the therapeutic effect of Liu Wei Di Huang Wan (LW) and its separated prescriptions, a classic traditional Chinese medicine formula for treating kidney yin deficiency in China. The study utilized ultra-performance liquid chromatography/electrospray ionization synapt high definition mass spectrometry (UPLC/ESI-SYNAPT-HDMS) in both negative and positive electrospray ionization (ESI). At the same time, blood biochemistry was examined to identify specific changes in the kidney yin deficiency. Distinct changes in the pattern of metabolites, as a result of daily administration of thyroxine and reserpine, were observed by UPLC-HDMS combined with a principal component analysis (PCA). The changes in metabolic profiling were restored to their baseline values after treatment with LW according to the PCA score plots. Altogether, the current metabonomic approach based on UPLC-HDMS and orthogonal projection to latent structures discriminate analysis (OPLS-DA) indicated 20 ions (14 in the negative mode, 8 in the positive mode, and 2 in both) as "differentiating metabolites".
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Pharmacokinetics-based elucidation on disparity in clinical effectiveness between varieties of Zhi Zhu Wan, a Traditional Chinese Medical formula.
J Ethnopharmacol
PUBLISHED: 01-28-2010
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Zhi Zhu Wan (ZZW) is a classical Chinese medical formulation used for the treatment of functional dyspepsia that attributed to Spleen-deficiency Syndrome. ZZW contains Atractylodes Rhizome and Fructus Citrus Immaturus, the later originates from both Citrus aurantium L. (BZZW) and Citrus sinensis Osbeck (RZZW). The present study is designed to elucidate disparities in the clinical efficacy of two ZZW varieties based on the pharmacokinetics of naringenin and hesperetin. MEHTOD: After oral administration of ZZWs, blood sample was collected from healthy volunteers at designed time points. Naringenin and hesperetin were detected in plasma by RP-HPLC, pharmacokinetic parameters were processed using mode-independent methods with WINNONLIN.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.