Publications by authors named "Qingqing Song"

52 Publications

Development and in vivo evaluation of MGF100-1R deletion mutant in an African swine fever virus Chinese strain.

Vet Microbiol 2021 Oct 14;261:109208. Epub 2021 Aug 14.

Shanghai Veterinary Research Institute, CAAS, Shanghai, 200241, China. Electronic address:

African swine fever virus (ASFV) is a large nucleoplasmic DNA virus, in which the genome is around 170-198 kilobases (kb). More than 50 % genes have unknown functions. Here, MGF100-1R gene is chosen to study the primary function and sublocalization. The gene was located at the left variable region of the ASFV genome that belongs to MGF100 families. It located at the cytoplasm without cytotoxic activities. However, it related to induce the transcriptional levels of pro-inflammatory cytokines. A deletion mutant of MGF100-1R gene was constructed based on ASFV Chinese strain GZ201801. The recombinant deletion mutant (ASFV△MGF100-1R) was demonstrated in vitro that the gene is non-essential for virus replication with a similar replication kinetics in bone marrow-derived macrophages (BMDMs) cell cultures when compared to parental virus. In vivo evaluation, ASFV△MGF100-1R was inoculated intramuscularly and led to a similar pathogenesis that caused by the parental ASFV GZ201801, confirming that deletion of MGF100-1R gene from the ASFV genome does not impact virulence.
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http://dx.doi.org/10.1016/j.vetmic.2021.109208DOI Listing
October 2021

Direct infusion-tandem mass spectrometry combining with data mining strategies enables rapid chemome characterization of medicinal plants: A case study of Polygala tenuifolia.

J Pharm Biomed Anal 2021 Sep 24;204:114281. Epub 2021 Jul 24.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China. Electronic address:

Data-independent MS spectrum acquisition after fragmenting the precursor ion cohort with 1 Da bin, termed as MS/MS, offers an opportunity to achieve rapid chemome characterization when being coupled with direct infusion (DI). Some post-acquisition data processing strategies, such as mass defect filtering (MDF), diagnostic fragment ion filtering (DFIF), and neutral loss filtering (NLF), facilitate data extraction from massive dataset, and moreover, molecular weight (MW) imprinting allows rapid capturing those reported components. Here, DI-MS/MS was employed to acquire cubic spectral dataset, and the strategies such as MW imprinting, MDF, DFIF, and NLF, were subsequently applied to filter the structural information. The integrated pipeline was utilized for the chemome characterization of Polygala tenuifolia, a famous edible medicinal plant. To aid information filtering, an in-house chemical library was built by comprehensively collecting structural information from some available databases. A single analytical run was completed within 5 min. For MS spectrum processing, MW imprinting was firstly applied to capture the compounds in the chemical library, and "five-point" MDF frames were employed to pursue saponins, oligosaccharide esters, and xanthones. Regarding MS spectral plot, DFIF and NLF were deployed to search information-of-interest. Structural identification was accomplished by carefully correlating precursor ions and MS spectra, applying the well-defined mass cracking rules, and referring to literature information as well as available databases. A total of 109 compounds, mainly saponins (40 ones), oligosaccharide esters (29 ones), and xanthones (19 ones), were captured and structurally annotated. MS spectra were also implemented for chemome comparison between Polygala tenuifolia and several similar plants belonging to Polygala genus, resulting in the observation of significant inter- and intra-species differences. Above all, DI-MS/MS is a promising choice for high-throughput chemome profiling of, but not limited to, medicinal plants, in particular when being integrated with post-acquisition data processing strategies.
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http://dx.doi.org/10.1016/j.jpba.2021.114281DOI Listing
September 2021

[Chemome profiling of Pien-Tze-Huang by online pressurized liquid extraction-ultra-high performance liquid chromatography-ion trap-time-of-flight mass spectrometry].

Se Pu 2021 May;39(5):478-487

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China.

Pien-Tze-Huang is one of the most famous traditional Chinese medicine prescriptions and consists of several precious medicinal materials, such as Notoginseng Radix et Rhizoma, Bovis Calculus, Snake Gall, and Moschus. However, its formula has not been completely revealed. It is mainly applied for the treatment of acute and chronic viral hepatitis, carbuncle, and boils caused by blood stasis, unknown swelling, bruises, and various inflammation disorders. The chemical composition of Pien-Tze-Huang is extremely complicated. Thus far, extensive attention has been paid to the principal chemical families in Pien-Tze-Huang, such as ginsenosides, bile acids, and muscone derivatives. Comprehensive chemical profiling, although of immense importance for systematic quality control, has not been achieved. Therefore, we configured a platform, namely online pressurized liquid extraction-ultra-high-performance liquid chromatography-ion trap-time-of-flight mass spectrometry (online PLE-UHPLC-IT-TOF-MS), to characterize the chemical profile of Pien-Tze-Huang in detail as well as to conduct source attribution, aiming to clarify the chemome of Pien-Tze-Huang and to provide a reliable method for quality assessment. A sub-microgram amount of Pien-Tze-Huang powder (0.3 mg) was placed in a hollow guard column, which was subsequently filled with clear silica gel. Filter membranes were used to seal the extraction vessel. The vessel was then placed in an adapted guard column holder and maintained in a thermal column oven (70 ℃). Metal tubing was used to connect the outlet of the guard column holder to the mass spectrometer. The extraction phase was maintained for 3 min by employing 0.1%(v/v) formic acid aqueous solution as the extraction solvent with a flow rate of 0.2 mL/min. Moreover, a six-port two-position electronic valve was introduced to automatically switch the system from extraction to elution phases. Within the elution phase, 0.1%(v/v) formic acid aqueous solution and acetonitrile composed the mobile phase, and the extracts were eluted with a gradient program. Because of the elevated temperature and pressure, the physical and chemical properties of water, especially polarity and solubility, were modified. Therefore, warm water could be an eligible green solvent to achieve wide polarity-spanned extraction. In addition, IT-TOF-MS was employed to acquire tandem mass spectrometry information. The mass fragmentation pathways of saponins and bile acids were carefully studied. Finally, according to authentic compounds, mass fragmentation pathways, reference information in the literature, and accessible databanks, a total of 73 signals were observed from Pien-Tze-Huang, of which 71 components were tentatively identified and assigned. Among them, 36 were from Notoginseng Radix et Rhizoma, 15 from Snake Gall, and 9 from Bovis Calculus, while the occurrences of the other 11 components were synergistically contributed by both Bovis Calculus and Snake Gall, through retrieving the in-house chemical database that was built by considering all accessible chemical information from Notoginseng Radix et Rhizoma, Bovis Calculus, Snake Gall, and Moschus. The other two compounds were assigned as unknown compounds. However, none of the components were assigned to Moschus because they mainly contained hydrophobic compounds, such as cycloketones, cholesterol, and sterols, among others, and it was difficult to detect them with the current measurement program. The extraction efficiency of online PLE was assessed by comparing it with the efficiency obtained from ultrasonication at the same time. According to base peak ion current chromatograms (BPCs) and mass spectrometry information, the efficiency of online PLE was greater than that of ultrasonic extraction, even through direct analysis. Online PLE-UHPLC-IT-TOF-MS is not only a tool fit for the concept of green analytical chemistry, but also a reliable analytical pipeline for the direct characterisation of other complicated matrixes. Above all, this study clarified the chemome of Pien-Tze-Huang and provided meaningful information for the quality control of this famous TCM prescription.
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http://dx.doi.org/10.3724/SP.J.1123.2020.10011DOI Listing
May 2021

Luteolin Ameliorates Experimental Pulmonary Arterial Hypertension via Suppressing Hippo-YAP/PI3K/AKT Signaling Pathway.

Front Pharmacol 2021 15;12:663551. Epub 2021 Apr 15.

Department of Cardiovascular Medicine, The Second Xiangya Hospital of Central South University, Hunan, China.

Luteolin is a flavonoid compound with a variety of pharmacological effects. In this study, we explored the effects of luteolin on monocrotaline (MCT) induced rat pulmonary arterial hypertension (PAH) and underlying mechanisms. A rat PAH model was generated through MCT injection. In this model, luteolin improved pulmonary vascular remodeling and right ventricular hypertrophy, meanwhile, luteolin could inhibit the proliferation and migration of pulmonary artery smooth muscle cells induced by platelet-derived growth factor-BB (PDGF-BB) in a dose-dependent manner. Moreover, our results showed that luteolin could downregulate the expression of LATS1 and YAP, decrease YAP nuclear localization, reduce the expression of PI3K, and thereby restrain the phosphorylation of AKT induced by PDGF-BB. In conclusion, luteolin ameliorated experimental PAH, which was at least partly mediated through suppressing HIPPO-YAP/PI3K/AKT signaling pathway. Therefore, luteolin might become a promising candidate for treatment of PAH.
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http://dx.doi.org/10.3389/fphar.2021.663551DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8082250PMC
April 2021

Integrated Strategy Drives Direct Infusion-Tandem Mass Spectrometry as an Eligible Tool for Shotgun Pseudo-Targeted Metabolomics of Medicinal Plants.

Anal Chem 2021 02 13;93(4):2541-2550. Epub 2021 Jan 13.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, East Road of North 3rd Ring, Chaoyang District, Beijing 100029, China.

Direct infusion (DI) has an extraordinary high-throughput advantage. Pseudo-targeted metabolomics (PTM) has been demonstrated integrating the merits of both nontargeted and targeted metabolomics. Herein, we attempted to implant DI into the PTM concept to configure a new strategy allowing shotgun PTM. First, a versatile MS/MSALL program was applied to acquire MS and MS spectra. Second, online energy-resolved MS (online ER-MS) was conducted to obtain breakdown graph as well as optimal collision energy (OCE) for each ion transition paired by precursor ion and the dominant product ion. Third, selected reaction monitoring (SRM) was responsible to output a quantitative dataset with a constant length. Moreover, breakdown graph also served as orthogonal structural evidence when matching MS spectra between DI-MS/MS and an in-house library to strengthen structural annotation confidence. To evaluate and illustrate the utility of the new strategy toward shotgun PTM of medicinal plants, in-depth chemome comparison was conducted within three species, all of which are edible medicinal plants and playing essential roles for turning the deserts into the oases. A total of 185 variables participated in the quantitative measurement program. Each diagnostic ion pair was featured with an OCE. Significant species differences occurred, and echinacoside, acteoside, isoacteoside, 2'-acetyl-acteoside, tubuloside B, mannitol, sucrose, betaine, malate, as well as choline were found to be confirmative chemical markers offering primary contributions toward the species discrimination. After cross-validation with LC-MS/MS, DI-MS/MS fortified with the new strategy is an eligible tool for shotgun PTM, beyond plants.
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http://dx.doi.org/10.1021/acs.analchem.0c04602DOI Listing
February 2021

Liquid chromatography-three-dimensional mass spectrometry enables confirmative structural annotation of cistanoside F metabolites in rat.

J Chromatogr B Analyt Technol Biomed Life Sci 2021 Jan 26;1162:122457. Epub 2020 Nov 26.

School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, PR China.

Clarification the existence forms, including prototype and metabolite(s) is the prerequisite for understanding in-depth the therapeutic mechanisms of a given agent, particularly when oral administration. However, it is still a long distance for unambiguous structural identification of metabolites even employing the cutting-edge MS/MS technique, and the determinant obstacle is produced by its inherent isomer-blind disadvantage. To tackle with this drawback, online energy-resolved mass spectrometry (online ER-MS) was introduced to enable isomeric discrimination after that high-resolution MS/MS provided empirical molecular formula as well as substructures. In-depth metabolic characterization of cistanoside F (CF), an effective natural product, was conducted as a proof-of-concept for the new strategy namely three-dimensional MS that was configured by MS, MS and online ER-MS as 1st, 2nd, and 3rd dimensions, respectively. Sensitive metabolite detection was assisted by predictive multiple-reaction monitoring function on Qtrap-MS, and the empirical formulas of all metabolites were calculated from the quasi-molecular ions yielded from IT-TOF-MS. Subsequently, substructures of each metabolite were constructed by combining the calculated element compositions and the well-defined mass fragmentation pathways. Finally, online ER-MS was responsible to generate optimal collision energies for bonds-of-interest, and enabled rational selection among candidate structures. A total of thirteen metabolites were detected and confirmatively identified in rat after oral treatment of CF using LC-3D MS. Acyl-migration, hydrolysis and sulfation played key roles for the metabolic fate of CF. More importantly, LC-3D MS is an eligible tool to achieve confidence-enhanced structural annotation of metabolites in biological matrices because of the unique isomeric differentiation ability from online ER-MS.
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http://dx.doi.org/10.1016/j.jchromb.2020.122457DOI Listing
January 2021

Confirmative Structural Annotation for Metabolites of ()-7,3'-Dihydroxy-4'-methoxy-8-methylflavane, A Natural Sweet Taste Modulator, by Liquid Chromatography-Three-Dimensional Mass Spectrometry.

J Agric Food Chem 2020 Nov 21;68(44):12454-12466. Epub 2020 Oct 21.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China.

Flavonoids occupy the largest family of natural products and possess a broad spectrum of health benefits. Their metabolites are sometimes the truly effective molecules . It is still challenging, however, to unambiguously identify flavonoid metabolites using conventional LC-MS/MS. Herein, we aimed to pursue auxiliary structural clues to / values in both MS and MS spectra through LC coupled to three-dimensional MS (LC-3D MS). MS, as the first dimension, was in charge of suggesting theoretical molecular formulas, MS, the as second dimension, was responsible for offering substructures, and exactly, online energy-resolved MS (ER-MS), as the third dimension, provided optimal collision energies (OCEs) that reflected the linkage manners among the substructures. Metabolic characterization of a natural sweet taste modulator, namely, ()-7,3'-dihydroxy-4'-methoxy-8-methylflavane (DHMMF), was conducted as a proof-of-concept. Extensive efforts, such as full MS and MS scans on IT-TOF-MS and predictive selected-reaction monitoring mode on Qtrap-MS, were made for in-depth metabolite mining. Seventeen metabolites (-) were captured from DHMMF-treated biological samples, including 17 (-), 10 (, , , , and ), and 2 ( and ) metabolites from urine, plasma, and feces, respectively. Their structures were configured by integrating MS, MS, and OCE information. Except , all metabolites were new compounds. LC-MS/MS-guided chromatographic purification yielded three glucuronyl-conjugated metabolites (, , and ), and NMR spectroscopic assays consolidated the structures transmitted from LC-3D MS. Demethylation, glucuronidation, and sulfation occurred as the primary metabolic pathways of DHMMF. Above all, LC-3D MS bridged LC-MS/MS from putatively structural annotation toward confidence-enhanced identification, beyond the metabolite characterization of flavonoids.
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http://dx.doi.org/10.1021/acs.jafc.0c05154DOI Listing
November 2020

[Direct chemome profiling of using online pressurized extraction-ultrahigh-performance liquid chromatography-ion trap-time-of-flight mass spectrometry].

Se Pu 2020 May;38(5):554-563

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China.

An online pressurized extraction-ultrahigh-performance liquid chromatography-ion trap-time-of-flight mass spectrometry (online PLE-UHPLC-IT-TOF-MS) platform was configured for the rapid and direct profiling of the chemical constituents of (). Notably, only a sub-microgram amount (0.5 mg) of was placed in an empty guard column core, which was then filled up with normal-phase silica gel as an extraction vessel. The guard column core was placed inside a guard column holder in the column oven (70℃) and connected to the UHPLC-IT-TOF-MS system via metal tubing. The extraction phase was maintained for 3 min with 0.1% (v/v) formic acid as the extraction solvent. The extraction and elution phases for the entire measurement were segmented by a 6-port/2-channel electronic valve. In the elution phase, 0.1% (v/v) aqueous formic acid-acetonitrile containing 0.1% (v/v) formic acid was the mobile phase, and the components of interest that accumulated at the front of the column were eluted into the IT-TOF-MS system for detection. A total of 48 compounds were observed, and according to the literature, database, and mass fragmentation pathways, 45 compounds, including 10 phenylethanol glycosides, 14 iridoid glycosides, and 21 phenylalanine glycosides, were identified from . This study provides reliable information regarding the chemical composition and quality evaluation of . Moreover, it offers a promising analytical pipeline for the chemical composition characterization of traditional Chinese medicines.
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http://dx.doi.org/10.3724/SP.J.1123.2019.10006DOI Listing
May 2020

Direct Infusion-Three-Dimensional-Mass Spectrometry Enables Rapid Chemome Comparison among Herbal Medicines.

Anal Chem 2020 06 18;92(11):7646-7656. Epub 2020 May 18.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China.

Direct infusion-mass spectrometry (DI-MS) currently serves as an alternative analytical tool for metabolomics owing to the unique high-throughput advantage. Except the inherent shortcoming at a significant matrix effect, there are two other primary technical obstacles dampening its wide applications, such as data alignment and structural annotation. To address these two obstacles, a novel strategy termed as DI-three-dimensional-MS (DI-3D-MS) was proposed here, and chemome comparison among several confusing herbal medicines (HMs) belonging to the family was conducted as a proof-of-concept. Each test sample was directly infused into Qtrap-MS. In the first dimension, stepwise multiple ion monitoring (MIM) program was implemented to universally acquire the quantitative information on all HMs and to generate aligned data files. In the second dimension, MS spectra were universally recorded by enhanced product ion (EPI) experiments that were triggered by MIM via an information-dependent acquisition algorithm. In the third dimension, online energy-resolved MS (ER-MS) was programmed to yield breakdown graphs for all MIM items. Moreover, a data library was built to aid structural identification by involving MS and CE features that were obtained by well-developed LC-MS methods. Qualitative and quantitative potentials of DI-3D-MS were validated toward metabolomics study. Significant species differences were observed, and all materials were grouped into three clusters. After matching MS spectra and breakdown graphs between DI-3D-MS and those in the data library, coumarins ubiquitously existed in each HM, and angular-type pyranocoumarins, linear-type pyranocoumarins, angular-type furanocoumarins, along with ligustilide derivatives offered primary contributions for the classification pattern. Above all, DI-3D-MS is an eligible choice for rapid metabolomics of HMs and other matrices as well.
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http://dx.doi.org/10.1021/acs.analchem.0c00483DOI Listing
June 2020

ECCM Schemes against Deception Jamming Using OFDM Radar with Low Global PAPR.

Sensors (Basel) 2020 Apr 7;20(7). Epub 2020 Apr 7.

National Demonstration Center for Experimental Electrical and Electronic Education, Yangtze University, Jingzhou 434023, China.

In this paper, a type of effective electronic counter-countermeasures (ECCM) technique for suppressing the high-power deception jamming using an orthogonal frequency division multiplexing (OFDM) radar is proposed. Concerning the velocity deception jamming, the initial phases of the pulses transmitted in a coherent processing interval (CPI) are designed to minimize the jamming power within a specific range, forming a notch around the jamming in the Doppler spectrum. For the purpose of suppressing the range deception jamming and the joint range-velocity deception jamming, the phase codes of the subcarriers belonging to the OFDM pulses are optimized to minimize the jamming power, distributing some specific bands in the range and the range-velocity domain, respectively. According to Parseval's theorem, the phase encoding, acting as the coding manner of the OFDM subcarriers can ensure that the energy of each OFDM symbol stays the same. It is worth noticing that the phase codes of the OFDM subcarriers can influence the peak-to-average power ratio (PAPR). Thus, an optimization problem is formulated to optimize the phase codes of the subcarriers under the constraint of global PAPR, which can regulate the PAPRs of multiple OFDM symbols at the same time. The proposed problem is non-convex; therefore, it is a huge challenge to tackle. Then we present a method named by the phase-only alternating direction method multipliers (POADMM) to solve the aforementioned optimization problem. Some necessary simulation results are provided to demonstrate the effectiveness of the proposed radar signaling strategy.
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http://dx.doi.org/10.3390/s20072071DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7181218PMC
April 2020

Family Management Style Improves Family Quality of Life in Children With Epilepsy: A Randomized Controlled Trial.

J Neurosci Nurs 2020 Apr;52(2):84-90

Introduction: Epilepsy is a common chronic disease with transient brain dysfunction and critically influences the quality of patients' family life. The aim of this study was to analyze the effectiveness of family management style on family quality of life in children.

Methods: We randomized 130 children to either the intervention group (n = 65) or the control group (n = 65). Family management style combined with routine care was applied in the intervention group within the first 24 hours after admission, whereas only routine care was applied in the control group. Family management style contains 3 steps: involve families into the intervention group and determine treatment plan, educate parents on how to manage their family, and monitor quality of home management. Scores on the Beach Center Family Quality of Life Scale (FQOL) of 2 groups were collected at 3 time points: within the first 24 hours after admission (T1), 6 months after discharge (T2), and 12 months after discharge (T3). Repeated-measures analysis of variance of FQOL scores was used to evaluate difference.

Results: Full scores and each subscale's scores on FQOL in the control group and the intervention group at T1 had no statistical significance (P > .05). Scores on FQOL at T2 and T3 increased in the intervention group, but there was almost no change in the control group, with statistical significance between the intervention group and the control group (P < .05). Scores on FQOL at T1, T2, and T3 showed that score of subscale except parenting FQOL improved in the intervention group (P < .05), but no difference was shown in the control group (P > .05). There was no difference shown among the control group and the intervention group that interacted with time (P > .05).

Conclusion: The family management style can effectively improve the family quality of life in children with epilepsy, especially at the satisfaction level of family emotional well-being and disability-related support.
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http://dx.doi.org/10.1097/JNN.0000000000000497DOI Listing
April 2020

Phenylethanol glycosides from Cistanche tubulosa improved reproductive dysfunction by regulating testicular steroids through CYP450-3β-HSD pathway.

J Ethnopharmacol 2020 Apr 24;251:112500. Epub 2019 Dec 24.

Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China; State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China. Electronic address:

Ethnopharmacological Relevance: Cistanche tubulosa (Schenk) R. Wight has been used frequently in traditional folk medicine for treatment of male sexual dysfunction (MSD). Phenylethanol glycosides, the main components of C. tubulosa, possess a variety of pharmacological activities due to their multiple properties. However, the underlying mechanism by which phenylethanol glycosides from C. tubulosa (CPhGs) regulates testicular steroids has not been elucidated to date.

Aim Of The Study: This study is to determine whether CPhGs promotes the reproductive functions of mice through CYP450-3β-HSD pathway of testosterone synthesis.

Materials And Methods: The major compositions of C. tubulosa (CPhGs) were quantified by high performance liquid chromatography (HPLC). The model of reproductive injury in mice were induced by injection of hydrocortisone (HCT). Different doses of CPhGs (72, 145 and 289 mg/kg) and testosterone propionate (TP, positive control drug) were administrated intragastrically for 14 d. The reproductive functions (erectile incubation period, capture and ejaculation incubation period, number of captures and ejaculations) and organ weights (testicle, epididymis, seminal vesicle and penis) were then determined. The levels of luteinizing hormone and testosterone in serum were quantified by radioimmunoassay. The key enzymes in testosterone synthesis pathways such as steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side chain cleavage enzyme (P450scc/CYP11A1) and 3β-hydroxysteroid dehydrogenase (3β-HSD) in the testis were assessed by immunofluorescence (IF) staining or/and Western blot (WB) analysis.

Results: The results illustrated that the low dose of CPhGs (72 mg/kg) had no significant protective effect against the reproductive injury caused by HCT, while the moderate dose of CPhGs (145 mg/kg) improved the damaged reproductive ability and the declined levels of luteinizing hormone and testosterone in the model mice (P < 0.001, P < 0.05, respectively). In particular, high dose of CPhGs (289 mg/kg) was most effective in improving HCT-induced changes in body weight (P < 0.01), reducing the incubation period of the erectile (P < 0.001), capture (P < 0.05) and ejaculation (P < 0.01), and increasing the number of captures and ejaculations (P < 0.01, P < 0.05, respectively). The weights of testcle, epididymis, seminal vesicle and penis (P < 0.001, P < 0.01, P < 0.01, P < 0.001, respectively) were improved by high dose of CPhGs. The levels of testosterone and its upstream luteinizing hormone were up-regulated by high dose of CPhGs (P < 0.001). Meanwhile, the expressions of the key steroidogenic enzymes including CYP11A1 and 3β-HSD were significantly up-regulated after CPhGs treatment (P < 0.001), demonstrated that CPhGs exerted the effect through enhancing testosterone biosynthesis via CYP450-3β-HSD pathway.

Conclusions: CPhGs could significantly protect against HCT-induced deleterious reproductive dysfunction and testis injury. The protective effects were exerted by up-regulating synthesis of testosterone via the CYP450-3β-HSD pathway in Leydig cells.
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http://dx.doi.org/10.1016/j.jep.2019.112500DOI Listing
April 2020

Genome-Wide Identification and Analysis of the Cytochrome B5 Protein Family in Chinese Cabbage ( L. ssp. ).

Int J Genomics 2019 2;2019:2102317. Epub 2019 Dec 2.

College of Life Science, Shandong Normal University, Jinan 250100, China.

Cytochrome B5 (CB5) family proteins play an important role in various oxidation/reduction reactions in cells as the electron donor and are involved in a variety of biotic and abiotic stress processes. However, the function of the in is still unclear. In this study, we carried out genome-wide identification, characterization, and expression analysis of in different tissues under adversities and stresses. It was identified that fifteen were distributed on different chromosomes, which were classified into seven groups (A-G) according to its phylogenetic relationship. Phylogenetic analysis of the CB5 protein sequences from six species showed that the BrCB5s conduct a close evolutionary process with the CB5s of and far from those of . Protein interaction analysis showed that 40 interaction patterns were predicted including two Sucrose Transporter 4 subfamily proteins (SUT 4) and Fatty Acid Hydroxylase 2 protein (FAH 2) can interact with most members of BrCB5s. The expression profile analysis indicated that were differentially expressed in different tissues, and the transcript abundances were significantly different under various abiotic stresses and plant hormone treatments. Our study provides a basis for a better understanding of the characteristics and biological functions of the CB5 family genes in Chinese cabbage during plant development, especially in plant responses to multiple stresses.
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http://dx.doi.org/10.1155/2019/2102317DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6913312PMC
December 2019

Retention Time and Optimal Collision Energy Advance Structural Annotation Relied on LC-MS/MS: An Application in Metabolite Identification of an Antidementia Agent Namely Echinacoside.

Anal Chem 2019 12 14;91(23):15040-15048. Epub 2019 Nov 14.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica , Beijing University of Chinese Medicine , Beijing 100029 , China.

The structural annotation of metabolites now relies heavily on HR-MS/MS information, resulting in ambiguous identities in most cases. More auxiliary evidence is therefore desired to achieve confirmative identification. Herein, we made an attempt to involve retention time () along with optimal collision energy (OCE) as the additionally structural clues, and the applicability validation was conducted via confidence-enhanced metabolite characterization of echinacoside, an antidementia drug candidate within clinical trials. Quantitative structure-retention relationships (QSRR) were modeled via assaying 184 authentic compounds on RPLC, HILIC, and serially coupled RPLC and HILIC (RPLC-HILIC). Online energy-resolved MS was developed to yield breakdown graphs for selected ion transitions, and OCE was demonstrated to be superior to CE toward pointedly denoting the bonds-of-interest. Nineteen metabolites (-) were confidently identified in biological samples from echinacoside-treated rats by analyzing / values first to yield empirical formulas and substructures, and and OCE subsequently contributed to sift the candidate structures. Structural identification was validated by oral administration of three relevant compounds in parallel and chromatographic purification as well. Above all, the integration of retention and dissociation behaviors enabled promoting one step forward for structural annotation confidences merely relied on HR-MS/MS.
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http://dx.doi.org/10.1021/acs.analchem.9b03720DOI Listing
December 2019

Binary code, a flexible tool for diagnostic metabolite sequencing of medicinal plants.

Anal Chim Acta 2019 Dec 20;1088:89-98. Epub 2019 Aug 20.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China. Electronic address:

The principle of chromatographic fingerprint is that certain diagnostic metabolites should be always distributed in a given plant and currently, it has been widely accepted as a promising means for medicinal plant authentication. Moreover, the chemical profile is the only evidence to clarify the ingredients of those consumable plant products, e.g. traditional Chinese medicine (TCM) prescriptions. Herein, efforts were made to describe the diagnostic metabolome of medicinal plant or TCM prescription using a binary code sequence. Forty-five well-known medicinal plants along with six relevant prescriptions were employed for concept illustration and proof. Each plant was subjected to chemical characterization, and diagnostic metabolites of all plants were gathered into a chemical pool containing 595 compounds. A robust method enabling the detection of all 595 constituents was then developed using LC coupled to scheduled multiple reaction monitoring. Analyst™ software was responsible for automatically judging the presence (defined as "1") or absence (defined as "0") of each analyte with a defined signal-to-noise threshold (S/N > 100). After converting each medicinal plant to a binary sequence consisting of 595 codes, an in-house database was built by involving all sequences. The potentials of sequence library retrieval towards plant authentication, preliminary chemical characterization, and deformulation of TCM prescriptions were demonstrated after that the diagnostic metabolome of each test sample was translated to a binary code sequence. Above all, binary code is a flexible tool for diagnostic metabolite sequencing of medicinal plants, and it should be an alternative tool of DNA barcoding towards plant authentication.
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http://dx.doi.org/10.1016/j.aca.2019.08.039DOI Listing
December 2019

Optimal collision energy is an eligible molecular descriptor to boost structural annotation: An application for chlorogenic acid derivatives-focused chemical profiling.

J Chromatogr A 2020 Jan 4;1609:460515. Epub 2019 Sep 4.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China. Electronic address:

Tandem mass spectrometry (MS/MS), in particular high-resolution MS/MS, is able to provide element compositions and substructures for the detected signals. However, it is still challenging to configure the whole structures via linking those substructures. Efforts were devoted here to propose and validate optimal collision energy (OCE) to be an auxiliary structural clue to mass-to-charge ratios (m/z), and online energy-resolved MS was developed to yield OCEs. Chlorogenic acid derivatives (CADs) were utilized as the proof-of-concept because diverse isomers usually initiated by the different linkage manners between the quinic acid/shikimic acid and cinnamoyl substituents(s), i.e. caffeoyl group, coumaroyl group, etc. Liquid chromatography-hybrid ion trap-time of flight MS (LC-IT-TOF-MS) was implemented to capture CADs in two well-known herbal medicines namely Lonicerae japonicae Flos and Inulae Flos. Afterwards, hybrid triple quadrupole-linear ion trap MS (Qtrap-MS) was deployed to acquire OCEs for the primary fragmentation pathways of all detected CADs through online energy-resolved MS. On the other side, structural calculations were conducted to figure out the relationships between OCEs and bond properties. Isomeric differences occurred for OCEs, and LC elution program as well as ionization parameters could not affect OCEs. Twenty-four and thirty-one CADs were hunted and putatively identified by LC-IT-TOF-MS in Lonicerae japonicae Flos and Inulae Flos, respectively, and the structural annotation was advanced by applying the OCE-bond property relationships. To verify the structures, CADs-of-interest were purified from Lonicerae japonicae Flos using an automated fraction collector and definitely identified with NMR spectroscopy. Exact consistence occurred for the structural identification of mono-caffeoylquinic acid isomers between LC-MS/MS and NMR analyses. Consequently, OCE is an inherent physicochemical parameter of a given compound and is an eligible structural descriptor to offset the ability of LC-MS/MS towards the chemical profiling of complex matrices.
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http://dx.doi.org/10.1016/j.chroma.2019.460515DOI Listing
January 2020

l-Glutamine Represses the Unfolded Protein Response in the Small Intestine of Weanling Piglets.

J Nutr 2019 11;149(11):1904-1910

State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing, China.

Background: Dysfunction of the endoplasmic reticulum (ER) results in apoptosis, inflammation, and enhanced proteolysis in the small intestine of humans and animals. l-Glutamine (Gln) is required for intestinal mucosal homeostasis in piglets. However, a functional role of the ER in the enterocytes of weanling piglets and its contribution to intestinal mucosal integrity remain largely unknown.

Objective: This study was conducted to test the hypothesis that preweaning administration of Gln alleviates the activation of unfolded protein response (UPR) in the small intestine of weanling piglets.

Methods: Eighteen sow-reared piglets aged 7 d from 3 litters (6 piglets/litter) were assigned randomly into 1 of 3 treatment groups. Piglets were reared by sows until age 24 d, or were reared by sows and orally administered either l-alanine [1.84 g · kg body weight (BW)-1 · d-1] or Gln (1.52 g · kg BW-1 · d-1) twice daily between 7 and 21 d of age, and then weaned to a corn- and soybean meal-based diet. The small-intestinal samples were collected at 24 d of age for analyses of abundance of proteins related to ER stress and apoptosis, concentrations of inflammatory cytokines, and mRNA abundance for genes implicated in protein degradation.

Results: Compared with age-matched suckling piglets, weaning stress increased apoptosis and decreased cell proliferation in the jejunum. The abundance of proteins related to ER stress [binding immunoglobulin protein, activating transcription factor 6α, phosphorylated (p)-inositol-requiring kinase 1α, and p-eukaryotic initiation factor 2α] was elevated by 200% to 320%, and that of apoptotic proteins (CCAAT/enhancer-binding protein homologous protein, p-Jun-N-terminal kinase, caspase-12, cleaved caspase-3, and Bcl-2-associated X) was augmented by 100% to 350% in the jejunum of weanling piglets. The protein abundance for IL-1β, TNF-α, and IL-8 was increased by 100% to 230% in the jejunum of weanling piglets. These alterations in gene and protein expression were markedly abrogated by Gln supplementation. The mRNA concentration of F-Box protein 32 in the jejunum of weanling piglets was increased by 70%, compared with the control group, and was not affected by Gln supplementation.

Conclusion: Our results indicate that preweaning administration of Gln to nursing piglets alleviates the weaning-activated UPR.
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http://dx.doi.org/10.1093/jn/nxz155DOI Listing
November 2019

The solvent effect on the photodeprotection of anthraquinone protected carboxylic acid unravelled by time-resolved spectroscopic studies.

Phys Chem Chem Phys 2019 Jul;21(27):14598-14604

Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong S.A.R., P. R. China.

Time-resolved spectroscopy studies coupled with the results from density functional theory (DFT) computations were utilized to unravel the photodeprotection reaction mechanism(s) of AQ protected p-methoxybenzoic acid (1) and its influence on the solvent was examined. It has been found that in aprotic organic solvents such as acetonitrile only the triplet state species was seen, whereas a hydrogen atom transfer (HAT) reaction takes place in isopropanol (IPA) to yield the aromatic ketyl radical species for 1. Compound 1 undergoes HAT and a proton transfer process sequentially to accomplish the release of the leaving group in protic organic solvent MeOH and MeOH-H2O solutions and photodeprotection is more favourable in the latter solution due to the stronger proton mediation ability of the water molecules and this system also facilitates the potential application of 1 in biological systems. Although the released acid from photodeprotection reaction can also be seen in another aqueous solvent system of THF-H2O, the by-product of the AQ chromophore undergoes side recombination reaction, which means this the solvent is not preferred for the photodeprotection process. This mechanistic work will help in the future design and development of AQ-PPGs for particular applications.
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http://dx.doi.org/10.1039/c9cp01227cDOI Listing
July 2019

Serial hyphenation of dried spot, reversed phase liquid chromatography, hydrophilic interaction liquid chromatography, and tandem mass spectrometry towards direct chemical profiling of herbal medicine-derived liquid matrices, an application in Cistanche sinensis.

J Pharm Biomed Anal 2019 Sep 22;174:34-42. Epub 2019 May 22.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China. Electronic address:

Inspired by dried blood spots (DBS), "dried spots of herbal medicines" (DSHM) concept was proposed here. In response to this superior sampling means, a new platform integrating dried spot, serially coupled reversed phase liquid chromatography and hydrophilic interaction liquid chromatography (RPLC-HILIC), and tandem mass spectrometry (MS/MS), was configured for directly, comprehensively chemical profiling of HM-derived liquid matrices. As an important original source of Cistanches Herba (Chinese name: Roucongrong) that is a well-known tonic HM, Cistanche sinensis (Csi) was employed to illustrate and validate the applicability. Dried spots (I.D.  3.0 mm) were prepared by loading 2 μL aliquots of Csi extract onto filter paper. Each dried spot was packed into an in-line filter holder (I.D. 3.0 mm × 4.0 mm) and then inserted behind the auto-sampler of a well-defined instrumentation named RPLC-HILIC-MS/MS. Hybrid ion trap-time of flight MS and hybrid triple quadrupole-linear ion trap MS were deployed in combination for the in-depth MS/MS data acquisition of diverse chemical families, such as phenylethanoid glycosides, lignans, iridoids, amino acids, and so forth. To assist chemical profiling, an in-house chemical library was built by collecting as much prior knowledge as possible. A total of 88 components were detected and tentatively annotated in Csi by matching their multi-stage MS spectra with those of authentic standards and literature data. Collectively, DSHM carried all merits of DBS, and the integrative dried spot-RPLC-HILIC-MS/MS platform was a promising analytical tool for direct chemical analysis and rapid quality evaluation of HMs, in particular those traditional Chinese medicine injections.
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http://dx.doi.org/10.1016/j.jpba.2019.05.048DOI Listing
September 2019

Advanced liquid chromatography-mass spectrometry enables merging widely targeted metabolomics and proteomics.

Anal Chim Acta 2019 Sep 11;1069:89-97. Epub 2019 Apr 11.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China.

Either widely targeted metabolomics or quantitative proteomics usually requires unique analytical platform. However, cross-platform omics studies entail higher levels of complexity and uncertainty, and result in a significant obstacle for high throughput assay as well. It is thereby urgent to pursue an integrative approach being capable of merging these two omics terms, namely widely targeted bi-omics. As an eligible analytical tool for large-scale targeted metabolomics, reversed phase liquid chromatography-hydrophilic interaction liquid chromatography-tailored selected reaction monitoring (RPLC-HILIC-tailored SRM) was deployed here to further receive the tryptic peptides as the analytes. Comparative evaluation of metabolites and tryptic peptides, 101 ones in total, between HepG2 and SK-Hep1 cells was conducted as a proof-of-concept. All analytes, regardless of metabolites or peptides, exhibited satisfactory chromatographic behaviors on RPLC-HILIC. Quantitative MS parameters, such as SRM transitions and collision energies (CEs), of either tryptic peptides or metabolites were online optimized in a standard compound-independent manner. It was worthwhile to mention that the signal responses of the peptides-of-choice generated by the optimized CEs were significantly superior to those values suggested by Skyline software. Calibration curves of both metabolites and peptides were constructed by serially diluting a so-called universal metabolome standard (UMS) sample. The quasi-content of each peptide or metabolite was gained according to applying those regressive calibration curves. After subjecting the quasi-content dataset into SIMCA-P software, significant differences took place between the two hepatic cell lines, and not only metabolites but tryptic peptides contributed to the discrimination. Above all, RPLC-HILIC-tailored SRM offered a promising choice towards widely targeted bi-omics attributing to the advantage of simultaneous monitoring metabolites and tryptic peptides.
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http://dx.doi.org/10.1016/j.aca.2019.04.013DOI Listing
September 2019

Toxicity studies for the use of prodrug of voriconazole in rats.

Regul Toxicol Pharmacol 2019 Jun 25;104:8-13. Epub 2019 Feb 25.

China Pharmaceutical University, Nanjing, 210009, China.

It has been reported that voriconazole is used to treat infections caused by invasive aspergillosis, fluconazole-resistant Candida, Actinoplanes and Fusarium. This study was performed to investigate the safety of prodrug of voriconazole (POV) and explore the distribution and metabolism of POV in vivo. The POV for injection was formulated into POV injection. In this study, POV injection was given intravenously at the doses of 0, 30, 60, and 120 mg/kg/d to SD rats for 4 weeks consecutively. Toxicokinetic study was also performed to explore its distribution and metabolism. POV injection was found to be safe and well tolerated. Some statistically significant differences in relative liver weight were observed and several cases of hepatocyte hypertrophy occurred after the 4-week POV injection treatment. Liver-related toxic response could be reversed after recovery period. The results of toxicokinetics showed that POV can rapidly converts to voriconazole in SD rats after administration. The exposure of voriconazole in each group was significantly different between male and female rats. The results showed that the target organ for the toxic effect of POV is liver and the no-toxic-reaction-dose for long-term administration of POV injection was 60 mg/kg/d.
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http://dx.doi.org/10.1016/j.yrtph.2019.02.013DOI Listing
June 2019

A random forest model to predict heatstroke occurrence for heatwave in China.

Sci Total Environ 2019 Feb 2;650(Pt 2):3048-3053. Epub 2018 Oct 2.

National Institute of Environmental Health, Chinese Center for Disease Control and Prevention, Beijing 100021, China. Electronic address:

Extreme heat events have recently become more frequent and represent an increasing risk of damage to public health. However, the existing prediction of heatwave related health effects has limited representativeness and verification. Our study addressed the prediction of heatstroke occurrences based on three years' data of typical cities of hot temperature in China, and examined the importance ranks of model parameters including meteorological and socioeconomic status (SES) factors. The results show that meteorological factors contributed the most to model estimation of the parameters evaluated, and SES parameters, such as the search index, were also important indicators of heatstroke prediction. The model had a satisfying performance compared to traditional linear regression models. The model established in our study can be further applied to extreme weather-related impact research and reduce economic loss due to public health expenses.
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http://dx.doi.org/10.1016/j.scitotenv.2018.09.369DOI Listing
February 2019

Unraveling the Photodeprotection Mechanism of Anthraquinon-2-ylmethoxycarbonyl-Caged Alcohols Using Time-Resolved Spectroscopy.

J Org Chem 2018 11 22;83(21):13454-13462. Epub 2018 Oct 22.

Department of Chemistry , The University of Hong Kong , Pokfulam Road , Hong Kong , P.R. China.

Anthraquinone (AQ) compounds have been used as photolabile protecting groups (PPGs) to protect alcohols, ketones, and carboxylic acids. However, because of the lack of direct spectroscopic information for the transient species and intermediates related with the deprotection reaction(s), the photorelease mechanism(s) of these systems are still largely unknown. In this contribution, we detail a time-resolved spectroscopic investigation using anthraquinone-protected galactose (1) and adenosine (2) to investigate the photodeprotection reaction mechanism(s) of these kinds of AQ-PPGs. It was found that, in THF-HO solvents, the ketyl radical species generated for 1 and 2 was found to be a reactive intermediate that then formed a dihydroxyanthracene species. The photodeprotection then occurred with the accompaniment of a decarboxylation process. These results provide an improved understanding for how AQ-PPGs work and will help assist in the design and applications of selected anthraquinone derivatives as a PPG platform, especially in aqueous environments more relevant for use in biological systems.
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http://dx.doi.org/10.1021/acs.joc.8b02252DOI Listing
November 2018

Serially coupled reversed phase-hydrophilic interaction liquid chromatography-tailored multiple reaction monitoring, a fit-for-purpose tool for large-scale targeted metabolomics of medicinal bile.

Anal Chim Acta 2018 Dec 15;1037:119-129. Epub 2017 Dec 15.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China. Electronic address:

The acquisition of high-quality quantitative dataset is the prerequisite for large-scale targeted metabolomics. However, the goal is usually dampened by the complexity of the biological matrices in terms of structural diversity, concentration span, and polarity range. We attempted herein to propose an analytical approach being able to circumvent these technical obstacles, and medicinal bile was employed as a proof of concept. In the liquid chromatography (LC) domain, reversed phase LC and hydrophilic liquid interaction chromatography were coupled in series, namely RPLC-HILIC, to yield appropriate chromatographic behavior for each component regardless of the polarity. In-depth chemical characterization and widely quantitative analysis were successively conducted in the mass spectrometry (MS) domain. Hybrid ion trap-time of flight MS was jointly deployed with hybrid triple quadrupole-linear ion trap MS for metabolite searching because of their orthogonal natures. Afterwards, a tailored MRM strategy that integrated online parameter optimization, ion intensity tailoring, and scheduled monitoring, was subsequently constructed to accomplish comprehensive quantitation although authentic compounds weren't involved and concentration span was quite large. Calibration curve was constructed for each detected identity by preparing and serially diluting the universal metabolome standard (UMS) solution that merged chemical information from all bile samples. Quasi-contents of 164 components including bile acids, sterols, eicosanoids, amino acids, lipids, flavonoids, and so forth, were gained by applying those regressive calibration curves and replaced the role of peak areas to serve as the variables for multivariate statistical analysis. In particular, the concentration ratio between tauroursodeoxycholic acid (TUDCA) and taurohyodeoxycholic acid (THDCA) that were exactly co-eluted as a single peak was calculated from the intensity specific value of m/z 498 > 80 over 498 > 124. Different metabolome occurred among various animal bile samples, and significant variations were also observed for different batches of bear bile powders. Bile acids acted as the indicative components in either discrimination. Overall, RPLC-HILIC-tailored MRM enabled quantitative metabolome profiling of medicinal bile and was thereby a promising analytical tool for large-scale targeted metabolomics.
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http://dx.doi.org/10.1016/j.aca.2017.11.072DOI Listing
December 2018

From H NMR-based non-targeted to LC-MS-based targeted metabolomics strategy for in-depth chemome comparisons among four Cistanche species.

J Pharm Biomed Anal 2019 Jan 6;162:16-27. Epub 2018 Sep 6.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China. Electronic address:

The great orthogonality between H NMR spectroscopy and LC-MS implies that their deployments in series could offer an opportunity to gain the qualified molecular markers via comparative metabolomics, and an attempt was made here to propose an integrated strategy namely "from H NMR-based non-targeted to LC-MS-based targeted metabolomics". In-depth chemome comparisons of Cistanche plants, such as C. deserticola, C. salsa, C. tubulosa, and C. sinensis, that possess dramatic economic and ecological benefits for the arid regions in the northwest China attributing to their dramatic medicinal and edible values, were employed to verify the applicability. H NMR-based non-targeted matabolomics acted as the survey experiment to find those signals offering decisive contributions towards the species discrimination, and the signals were translated to a set of putative identities, eighteen ones in total, through matching with authentic compounds and referring to some accessible databases. Afterwards, an advanced LC-MS platform assembling reversed phase liquid chromatography, hydrophilic interaction liquid chromatography, and tailored multiple reaction monitoring, was introduced to simultaneously quantify those eighteen potential markers in a single analytical run, because those candidates exhibited great polarity span as well as wide content range. Significant species differences occurred amongst their chemome patterns. Echinacoside, acteoside, betaine, mannitol, 6-deoxycatalpol, sucrose, and 8-epi-loganic acid were disclosed as the markers enabling the discrimination of those four species. The findings offered an alternative tool to differentiate Cistanche plants. More importantly, the strategy namely "from H NMR-based non-targeted to LC-MS-based targeted metabolomics" facilitates the pursuit of molecular markers among analogue plants, and thereby provides a promising choice for in-depth chemome comparison.
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http://dx.doi.org/10.1016/j.jpba.2018.09.013DOI Listing
January 2019

Authentic compound-free strategy for simultaneous determination of primary coumarins in Peucedani Radix using offline high performance liquid chromatography-nuclear magnetic resonance spectroscopy-tandem mass spectrometry.

Acta Pharm Sin B 2018 Jul 21;8(4):645-654. Epub 2018 Feb 21.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China.

Herein, a strategy is proposed for the simultaneous determination of primary coumarins in Peucedani Radix (Chinese name: Qianhu). The methodology consists of three consecutive steps: 1) Semi-preparative LC in combination with a home-made automated fraction collection module to fragment the universal metabolome standard into ten fractions (Frs. I-X); 2) LC-accurate MS/MS and quantitative H NMR spectroscopy conducted in parallel to acquire the qualitative and quantitative data of each fraction; 3) Robust identification and quantification of components by use of LC coupled to multiple reaction monitoring. In this final step, the most significant fractions (Frs. III-X) were pooled to serve as the pseudo-mixed standard solution. Meticulous online parameter optimization was performed to obtain the optimal parameters, including ion transitions and collision energies. Concerns were particularly paid onto pursuing the parameters being capable of monitoring -specific isomers, notably praeruptorin E 3'-isovaleryl-4'-angeloylkhellactone. The quantitative performance of the method was validated according to diverse assays. Eleven primary coumarins (-) were unambiguously identified and absolutely quantified, even though no external reference compound was used. Above all, the integrated strategy not only provides a feasible pipeline for the quality assessment of Peucedani Radix, but more importantly, shows the potential for authentic compound-free quantitative evaluation of traditional Chinese medicines.
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http://dx.doi.org/10.1016/j.apsb.2018.01.005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6090009PMC
July 2018

Integrated approach for confidence-enhanced quantitative analysis of herbal medicines, Cistanche salsa as a case.

J Chromatogr A 2018 Aug 22;1561:56-66. Epub 2018 May 22.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China. Electronic address:

Although far away from perfect, it is practical to assess the quality of a given herbal medicine (HM) through simultaneous determination of a panel of components. However, the confidences of the quantitative outcomes from LC-MS/MS platform risk several technical barriers, such as chemical degradation, polarity range, concentration span, and identity misrecognition. Herein, we made an attempt to circumvent these obstacles by integrating several fit-for-purpose techniques, including online extraction (OLE), serially coupled reversed phase LC-hydrophilic interaction liquid chromatography (RPLC-HILIC), tailored multiple reaction monitoring (MRM), and relative response vs. collision energy curve (RRCEC) matching. Confidence-enhanced quantitative analysis of Cistanche salsa (Csa), a well-known psammophytic species and tonic herbal medicine, was conducted as a proof-of-concept. OLE module was deployed to prohibit chemical degradation, in particular E/Z-configuration transformation for phenylethanoid glycosides. Satisfactory retention took place for each analyte regardless of polarity because of successive passing through RPLC and HILIC columns. Optimum parameters for the minor components, at the meanwhile of inferior ones for the abundant ingredients, ensured the locations of all contents in the linear ranges. The unequivocal assignment of the captured signals was achieved by matching retention times, ion transitions, and more importantly, RRCECs between authentic compounds and suspect peaks. Diverse validation assays demonstrated the newly developed method to be reliable. Particularly, the distribution of mannitol rather than galactitol was disclosed although these isomers showed identical retention time and ion transitions. The contents of 21 compounds-of-interest were definitively determined in Csa as well as two analogous species, and the quantitative patterns exerted great variations among not only different species but different Csa samples. Together, the fortification of OLE-RPLC-HILIC-tailored MRM with RRCEC matching could fully address the demands from confidence-enhanced quantitative analysis of HMs.
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http://dx.doi.org/10.1016/j.chroma.2018.05.045DOI Listing
August 2018

R-lipoic acid overdosing affects platelet life span via ROS mediated autophagy.

Platelets 2018 Nov 6;29(7):695-701. Epub 2017 Oct 6.

a Jiangsu Center for Safety Evaluation of Drugs, Jiangsu Province Institute of Materia Medica, School of Pharmaceutical Sciences, Nanjing Tech University , Nanjing 211800 , China.

R-lipoic acid (ALA), a powerful antioxidant valuable for the treatment of diabetes and its complications, has been reported to exhibit an antiplatelet activity in vitro. The aim of this study was to investigate the effect and mechanism of ALA on platelets in vivo. Sprague-Dawley (SD) male rats were intravenously administered with low-dose ALA (20 mg/kg/d), high-dose ALA (80 mg/kg/d) and saline, respectively. Platelets count and bone marrow smear were evaluated and the expressions of markers related to apoptosis and autophagy were measured. Platelet clearance analysis was conducted out on mice. The results showed that high-dose ALA administration could significantly decrease platelet count by 43% compared with control group, whereas, megakaryocytes showed no difference in the number. Moreover, high-dose ALA administration led to significant reduction in half-life of circulating platelets, indicative of enhanced rate of platelet clearance. Interesting, high-dose ALA administration could increase the level of reactive oxygen species (ROS) in platelets and induce autophagy without affecting apoptosis. Our finding also showed that high ALA-induced autophagy in platelets was mediated by class III PtdIns3K activity, which could be reversed by 3-methyladenine (3-MA). Moreover, AKT and MAPK/ERK pathways were also observed to be involved in the regulation of autophagy in platelets. Thus, high-dose ALA could induce autophagy in platelets through modulating the activity of class III PtdIns3K, which was associated with decreased count of circulating platelets and shortened lifespan of platelets.
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http://dx.doi.org/10.1080/09537104.2017.1356450DOI Listing
November 2018

Time-Resolved Spectroscopic Study on the Photoredox Reaction of 2-(p-Hydroxymethyl)phenylAnthraquinone.

Sci Rep 2017 08 22;7(1):9154. Epub 2017 Aug 22.

Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong S.A.R., P. R. China.

In this work, we report a combined time-resolved spectroscopic and density functional theory computational study on 2-(p-hydroxymethyl)phenylanthraquinone (PPAQ) in which the benzyl alcohol moiety is significantly farther away from the AQ ketone group compared to the compound 2-(1-hydroxyethyl) 9,10-anthraquinone (HEAQ) so as to investigate the photophysical and photochemical reactions of PPAQ in several solvents, especially for the photoredox reaction in a pH 2 aqueous solution. The results here indicate that PPAQ undergoes the photoredox reaction via a two-step pathway and that the low efficiency of the photoredox reaction of PPAQ compared to the related HEAQ molecule is caused by the longer distance between the benzyl alcohol moiety and the AQ ketone moieties.
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http://dx.doi.org/10.1038/s41598-017-09192-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5567363PMC
August 2017

Simultaneous determination of components with wide polarity and content ranges in Cistanche tubulosa using serially coupled reverse phase-hydrophilic interaction chromatography-tandem mass spectrometry.

J Chromatogr A 2017 Jun 15;1501:39-50. Epub 2017 Apr 15.

Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China. Electronic address:

To meet the demands from plant physiologists and pharmacognosists, sustainable efforts are being devoted by the analytical chemists from all over the world to search an approach being capable of simultaneously monitoring primary along with secondary metabolites. The key technical bottlenecks currently lie at affording satisfactory chromatographic and spectrometric performances for both hydrophilic and hydrophobic substances that span a great content range. Herein, reverse phase liquid chromatography was directly coupled with hydrophilic interaction chromatography, namely RPLC-HILIC, to integrate their merits, whereas dilution pumps were employed to tackle the mismatching for the mobile phase between them. On the other side, inferior parameters rather than the optimal ones were applied for those abundant ingredients to advance the upper limits of quantitation, such as echinacoside (1250.0μg/mL), mannitol (100.0μg/mL), and acteoside (125μg/mL), in mass spectrometer domain. As a desert parasitic plant as well as tonic materials, Cistanche tubulosa (CT) has drawn widely interests from plant physiologists and pharmacognosists regarding its quantitative metabolome. Simultaneous determination of 23 abundant and minor ingredients covering most chemical families in CT, i.e. amino acid, nucleoside, organic acid, phenylethanoid glycoside, lignan, and iridoid, was attempted to understand the physiologic patterns as well as pharmacological values of the crude materials. Although the analytes spanned wide polarity (-3.326≤cLogP≤1.421) and content (more than 5 orders of magnitude) ranges, satisfactory chromatographic and spectrometric behaviors were gained for all analytes. Reliable quantitation was demonstrated via various method validation assays, such as recovery, linearity, sensitivity, and precision. The contents of 23 hydrophilic and hydrophobic substances were quantified in 20 batches of CT. Significant variations occurred for the contents of those components. Echinacoside (1.35-387.50mg/g) was usually observed as the most abundant component, whereas ferulic acid (<0.0043mg/g) always exhibited trace distributions. Above all, the integrated equipment setup could serve as a fit-for-purpose tool for in-depth quality evaluation of CT and more importantly, for comprehensively understanding the metabolome of plants.
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http://dx.doi.org/10.1016/j.chroma.2017.04.034DOI Listing
June 2017
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