Systematic screening and structural characterization of dipeptides using offline 2D LC-LTQ-Orbitrap MS:A case study of Cordyceps sinensis

Cordyceps sinensis(C.sinensis)is a widely used and highly valuable traditional Chinese medicine.Several dipeptides have been detected in C.sinensis,but current scientific knowledge of its chemical makeup remains limited.In this study,an improved approach that integrates offline two-dimensional liquid chromatography(2D LC)separation,precursor ion list,library screening,and diagnostic ion filtering was established to systematically screen and characterize dipeptides in C.sinensis.Offline 2D LC integrating hydrophilic interaction LC and reverse phase separations was established to eliminate interference and identify the target dipeptides.A library containing the potential 400 dipeptides was created,and a precursor ion list with all theoretical precursor ions was adopted to trigger the MS/MS scan with high sensitivity.To identify dipeptides,the type and connection sequence of amino acids were determined according to the product ions.Ile and Leu residues were differentiated for the first time according to the characteristic ion at m/z 69.07.Ultimately,170 dipeptides were identified or tentatively characterized from C.sinensis,and most are reported for the first time in this species herein.In addition,the identified dipeptides were also applied for discrimination among the three Cordyceps species,and 11 markers were identified.The obtained results provide a deeper understanding of the chemical basis of C.sinensis.

Water extracts of Cordyceps sinensis inhibits proliferation and metastasis via regulating cell cycle and matrix metalloproteinases in melanoma

Background:Melanoma has poor survival rate owing to its aggressiveness and high rates of metastasis and recurrence.Only few effective therapies are available for metastatic melanoma,so,development of novel anti-metastatic drugs is in urgent need.Cordyceps sinensis is a well-known and common Chinese medicine and has been used as anti-cancer adjuvant treatments for many years.In our study,we aim to evaluate the effects of water extracts of Cordyceps sinensis(WECS)in A375 melanoma cells and in zebrafish model of melanoma metastasis.Methods:Initially,we detected the effect of WECS on the viability in A375 melanoma cells by MTT assay and colony formation assay,and then we detected the effect of WECS on adhesion and metastasis by wound healing assay and Transwell chambers assays.Next,we also explored underlying mechanism by Western blot.Notably,in vivo anti-metastasis assays with A375 zebrafish xenograft model also were performed.Results:As expected,our results found that WECS suppressed proliferation,adhesion and metastasis in A375 melanoma cells via regulating AKT/MMP2/MMP9 pathways and cell cycle.Furthermore,we also confirmed anti-metastasis efficacy of WECS with A375 zebrafish xenograft model.Conclusion:Overall,our results indicate that WECS,as a kind of metastasis inhibitor,may be developed as a new therapeutic strategy for metastatic melanoma treatment.

Profiling and identification of aqueous extract of Cordyceps sinensis by ultra-high performance liquid chromatography tandem quadrupole-orbitrap mass spectrometry

Characterization of aqueous extract in traditional Chinese medicine(TCM) is challenging due to the poor retention of the analytes on conventional C18 columns. This study presents a systematic characterization method based on a rapid chromatographic separation(8 min) on a polar-modified C18(Waters Cortecs T3) column of aqueous extract of Cordyceps sinensis. UHPLC-HRMS method was used to profile components in both untargeted and targeted manners by full MS/PIL/dd-MS2 acquisition approach. The components were identified or tentatively identified by reference standards comparison, fragmentation rules elucidation and available databases search. A total of 91 components, including 10 nucleobases, 20 nucleosides, 39 dipeptides, 18 amino acids and derivatives and 4 other components, were characterized from the aqueous extract of C. sinensis. And this was the first time to systematically report the presence of nucleosides and dipeptides in C. sinensis, especially for modified nucleosides. The chemical basis inquiry of this work would be beneficial to mechanism exploration and quality control of C. sinensis and related products. Meanwhile, this work also provided an effective solution for characterization of aqueous extract in TCM.

Discovery of differential sequences for improving breeding and yield of cultivated Ophiocordyceps sinensis through ITS sequencing and phylogenetic analysis

To accelerate the breeding process of cultivated Ophiocordyceps sinensis and increase its yield,it is important to identify molecular fingerprint of dominant O.sinensis.In the present study,we collected 3 batches of industrially cultivated O.sinensis product with higher yield than the others and compared their internal transcribed spacer(ITS)sequences with the wild and the reported.The ITS sequence was obtained by bidirectional sequencing and analyzed with molecular systematics as a DNA barcode for rapid and accurate identification of wild and cultivated O.sinensis collected.The ITS sequences of O.sinensis with detailed collection loci on NCBI were downloaded to construct a phylogenetic tree together with the sequences obtained from the present study by using neighbor-joining method based on their evolution relationship.The information on collection loci was analyzed with ArcGIS 10.2 to demonstrate the geographic distribution of these samples and thus to determine the origin of the dominant samples.The results showed that all wild and cultivated samples were identified as O.sinensis and all sequences were divided into seven phylogenetic groups in the tree.Those groups were precisely distributed on the map and the process of their system evolution was clearly presented.The three cultivated samples were clustered into two dominant groups,showing the correlation between the industrially cultivated samples and the dominant wild samples,which can provide references for its optimized breeding in the future.

Developmental recording of the ghost-moth larvae after ex situ infection by Ophiocordyceps sinensis

Dear Editor,Chinese cordyceps, a valuable Chinese traditional medicine, is a complex of mummified ghost moth caterpillars and fungal stroma of Ophiocordyceps sinensis (Winkler, 2010). O. sinensis has been nominated as the national fungus of China and flagship species of the Tibetan Plateau (Cannon, 2010;Zhang et al., 2012).

Analysis of Cordyceps by multi-column liquid chromatography

Cordyceps is a famous traditional Chinese medicine (TCM) that has been used in China for hundreds of years. In the present study a multi-column liquid chromatography (MC-LC) system was developed for the qualitative analysis of macromolecules and micromolecules in Cordyceps. The MC-LC system includes a size exclusion pre-column, a size exclusion column (SEC) and a reversed phase column (RP) which were controlled by column-switching valves. The sample was separated by the size exclusion pre-column into two fractions (macromolecules and micromolecules). These fractions were further separated on SEC and RP columns, respectively. A diode array detector (DAD) and a mass spectrometer (MS) were used to detect the components. This MC-LC method was utilized for analysis of Cordyceps samples. Two macromolecular peaks and 15 micromolecular peaks were found in Cordyceps, and 11 of the micromolecular peaks were identified as adenosine-5'-monophosphate (AMP), phenylalanine, uridine, hypoxanthine, inosine, guanine, guanosine, deoxyadenosine-5'-monophosphate (dAMP), adenosine, adenine and cordycepin (or its isomer). This method is useful for quality control of Cordyceps. (C) 2017 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V.

Efficacy and Safety of All-oral Emitasvir and Sofosbuvir in Patients with Genotype 1b HCV Infections without Cirrhosis

Publications>Journals>Journal of Clinical and Translational Hepatology>Article Full Text ORIGINAL ARTICLE OPEN ACCESS Efficacy and Safety of All-oral Emitasvir and Sofosbuvir in Patients with Genotype 1b HCV Infections without Cirrhosis Huiying Rao1,Xingxiang Yang2,Youwen Tan3,Qin Ning4,Daokun Yang5,Jiefei Wang6,Yongfeng Yang7,Sujun Zheng8,Dongliang Yang9,Jinlin Hou10,Qing Xie11,Caiyan Zhao12,Lunli Zhang13,Xiaorong Mao14,Tong Sun15,Lang Bai16,Fuchun Zhang17,Jinglan Jin18,Yingren Zhao19,Maorong Wang20,Wen Xie21,Yingjie Ma22,Jun Quan23,Xuebing Yan24,Ping An25,Feng Lin26,Jidong Jia27,Xiaoxuan Hu28,Zuojiong Gong29,Jie Wu30,Yongping Chen31,Zhansheng Jia32,Minghua Lin33,Guiqiang Wang34,Yueyong Zhu35,Yingjun Zhang*,36,Hongming Xie36,Lin Luo36,Qingyun Ren36,Rui Huang1 and Lai Wei*,37 Author information Journal of Clinical and Translational Hepatology 2020;8(3):255-261DOI:10.14218/JCTH.2020.00031 Abstract Background and Aims:Emitasvir is a new type of hepatitis C virus(HCV)nonstructural protein 5A(NS5A)inhibitor,and the data of phase 2 trial has shown emitasvir-sofosbuvir to have good safety and tolerance.We conducted this phase 3 trial to further verify the efficacy and safety.Methods:We evaluated the antiviral activity and safety of a 12-week regimen of emitasvir phosphate(100 mg)combined with sofosbuvir(400 mg)once daily in non-cirrhotic patients with genotype 1 HCV infection.The primary endpoint was a sustained virological response at 12 weeks(SVR12)after the end of treatment.Results:Of the 362 patients enrolled in the trial,39.8%were male,99.2%had HCV genotype 1b,0.8%had genotype 1a and 79.8%were treatment-naïve.The average age was 47.2 years.All patients completed the treatment and follow-up.All 3 patients with genotype 1a achieved SVR.Two genotype 1b treatment-naïve patients experienced virologic relapse.The rate of SVR12 was 99.7%(358/359),and SVR24 was 99.4%(357/359)in genotype 1b.Overall,36.2%had resistance-associated substitutions(RASs)in NS5A and 98.3%had RASs in NS5B at baseline.The RASs at baseline had no effect on the rates of response.Serious adverse events were reported in 16 patients and were not related to emitasvir-sofosbuvir.Most adverse events did not require therapy.Conclusions:The 12 weeks of treatment with emitasvir-sofosbuvir was a highly efficient and safe treatment for a wide range of patients with HCV genotype 1b infection without cirrhosis,who had not been treated or who had been treated with interferon-based regimen previously.

Nanostructured boron agents for boron neutron capture therapy:a review of recent patents

Boron neutron capture therapy(BNCT)is a potential radiation therapy modality for cancer,and tumortargeted stable boron-10(10B)delivery agents are an important component of BNCT.Currently,two low-molecular-weight boron-containing compounds,sodium mercaptoundecahydrocloso-dodecaborate(BSH)and boronophenylalanine(BPA),are mainly used in BNCT.Although both have suboptimal tumor selectivity,they have shown some therapeutic benefit in patients with high-grade glioma and several other tumors.To improve the efficacy of BNCT,great efforts have been devoted for the development of new boron delivery agents with better uptake and favorable pharmacokinetic profiles.This article reviews the application and research progress of boron nanomaterials as boron carriers in boron neutron capture therapy and hopes to stimulate people’s interest in nanomaterial-based delivery agents by summarizing various kinds of boron nanomaterial patents disclosed in the past decade.

Erratum to:Developmental recording of the ghost-moth larvae after ex situ infection by Ophiocordyceps sinensis

This paper contains errors in the authors’affiliations.Wenjia Li,is a Ph.D.candidate in the University of Chinese Academy of Sciences,Jiangchun Wei and Xingzhong Liu are professors in the University of Chinese Academy of Sciences,which is missing in the original version.