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Zhang M.,State Key Laboratory for Infectious Disease Prevention and Control China CDC | Zhang M.,Chinese National Institute for Communicable Disease Control and Prevention | Zhang M.,Beijing Institute of Technology | Zhang M.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | And 9 more authors.
Analytical Methods | Year: 2014

The CP4 EPSPS gene is widely used in herbicide-tolerant crops/plants all over the world. In this study, a sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed and validated to quantify the amount of CP4 EPSPS expression in Nicotiana tabacum leaves. The quantification of protein was used to measure the unique peptides of the CP4 EPSPS protein. Two peptides unique to CP4 EPSPS were synthesized and labelled in H 2 18O to give 18O stable isotope labelled peptides which served as internal standards. The validated method resulted in good specificity and linearity. The intra- and inter-day precisions and accuracy for all samples were satisfactory. The results demonstrated that the novel method was sensitive and selective to quantify CP4 EPSPS in the crude extract without time-consuming pre-separation or purification procedures. © the Partner Organisations 2014.


Wang L.,Beijing 302 Hospital | Sun Y.,Beijing 302 Hospital | Zhang Z.,Beijing 302 Hospital | Jia Y.,Beijing 302 Hospital | And 14 more authors.
Hepatology | Year: 2015

There is increasing interest in the role of T follicular helper (Tfh) cells in autoimmunity from the perspective of both their role in breach of tolerance and their effects on the natural history of disease progression. Indeed, the critical role of Tfh cells in autoimmunity is further highlighted based on their location in the germinal center (GC), a pathogenic hot spot for development of autoreactivity. To address the role of Tfh cells in primary biliary cirrhosis (PBC), we comprehensively evaluated the immunobiology of CXCR5+CD4+ Tfh cells in 69 patients with PBC, including a nested subgroup of 16 autoimmune hepatitis (AIH) and 20 healthy controls (HC), followed for 1 year. We report herein several key observations. First, there was an increased frequency of circulating Tfh cells in patients with PBC compared to AIH (P<0.05) and HC (P<0.01). Second, the function of circulating Tfh cells from PBC patients, including interleukin (IL)-21 production (P<0.05), the ability to promote B-cell maturation, and autoantibody production, were greater than HC. Third, the frequency of these cells was significantly decreased in ursodeoxycholic acid (UDCA) responders compared to UDCA-treated nonresponders, in both cross-sectional (P=0.023) and longitudinal studies (P=0.036), respectively. Indeed, similar increases of Tfh cells were noted in liver and spleen. Conclusion: These results significantly extend our understanding of lymphoid subpopulations in PBC and their relative role in disease expression. Our data also provide a novel biomarker for evaluation of the effectiveness of new therapeutic approaches. © 2014 by the American Association for the Study of Liver Diseases.


Liu Q.,First Affiliated Hospital | Qian Y.,First Affiliated Hospital | Chen F.,First Affiliated Hospital | Chen F.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | And 6 more authors.
Acta Biochimica et Biophysica Sinica | Year: 2014

Endotoxin lipopolysaccharide (LPS) plays an important role in the acceleration of inflammatory reaction of hepatitis as the second attack. Compounds that can prevent inflammation by targeting LPS have potential therapeutic clinical application. Epigallocatechin-3-gallate (EGCG) has potent hepatocyte-protective effect and mild anti-hepatitis virus function. Here, we investigated whether EGCG attenuated the severity of inflammatory response in LPS-stimulated L02 hepatocytes. L02 hepatocytes were pretreated with EGCG for 2 h, then stimulated by LPS at 250 ng/ml. The expression levels of chemokine regulated upon activation normal T-cell expressed and secreted (Rantes) and monocyte chemotactic protein-1 (MCP-1), pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interferon-γ, adhesion molecule intercellular adhesion molecule-1 (ICAM-1), oxidant stress molecules nitric oxide (NO), vascular endothelial growth factor (VEGF), and matrix metalloproteinase-2 (MMP-2) were tested by enzyme-linked immunosorbent assay. The expression of total extracellular signal-regulated kinase 1/2 (ERK1/2), phospho-ERK1/2 (p-ERK1/2), p-AKT, total p38, phospho-p38 (p-p38), total p65 and phospho-p65 (p-p65), IκBα, phospho-IκBα (p-IκBα) and TNF receptor associated factor 2 were tested by western blot analysis. Our results showed that pre-treatment with EGCG could significantly reduce the production of TNF-α, Rantes, MCP-1, ICAM-1, NO, VEGF, and MMP-2 in LPS-stimulated L02 hepatocytes in a dose-dependent manner. The effect of EGCG may be related to the inhibition of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways by down-regulation of p-IκBα, p65, p-p65, p-p38, p-ERK1/2, and p-AKT. These results indicate that EGCG suppresses LPS-induced inflammatory response and oxidant stress and exerts its hepatocyte-protective activity partially by inhibiting NF-κB and MAPK pathways. © 2013 The Author.


Mao L.,Centers for Disease Control and Prevention | Zhang E.,Chinese National Institute for Communicable Disease Control and Prevention | Zhang E.,State Key Laboratory for Infectious Disease Prevention and Control | Zhang E.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | And 23 more authors.
PLoS ONE | Year: 2016

Anthrax is a continuous threat in China, especially in rural regions. In July 2015, an anthrax outbreak occurred in Xifeng County, Liaoning Province. A total of 10 cutaneous anthrax cases were reported, with 210 people under medical observation. In this study, the general characteristics of human anthrax outbreak occurred in Liaoning Province were described, and all cases were caused by butchering and contacting sick animal. Meanwhile, the phylogenetic relationship between outbreak-related isolates/samples of the year 2015 and previous Bacillus anthracis strains was analyzed by means of canonical single nucleotide polymorphisms (canSNP), multiple-locus variable-number tandem repeat analysis (MLVA) with 15 markers and single-nucleotide repeats (SNR) analysis. There are two canSNP subgroups found in Liaoning, A.Br.001/002 and A.Br.Ames, and a total of six MLVA 15 genotypes and five SNR genotypes were observed. The strain collected from anthrax outbreak in Xifeng County in 2015 was classified as A.Br.001/002 subgroup and identified as MLVA15-29 genotype, with same SNR profile (CL10: 17, CL12: 15, CL33: 29, and CL35: 13). So we conclude that the same clone of B.anthracis caused the anthrax outbreak in Xifeng County in 2015, and this clone is different to previous isolates. Strengthening public health education in China is one of the most important measures to prevent and control anthrax. © 2016 Mao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Ribas de Pouplana L.,Barcelona Institute for Research in Biomedicine | Ribas de Pouplana L.,Catalan Institution for Research and Advanced Studies | Santos M.A.S.,University of Aveiro | Zhu J.-H.,Tsinghua University | And 3 more authors.
Trends in Biochemical Sciences | Year: 2014

The translation of genes into functional proteins involves error. Mistranslation is a known cause of disease, but, surprisingly, recent studies suggest that certain organisms from all domains of life have evolved diverse pathways that increase their tolerance of translational error. Although the reason for these high error rates are not yet clear, evidence suggests that increased mistranslation may have a role in the generation of diversity within the proteome and other adaptive functions. Error rates are regulated, and there appears to be an optimal mistranslation rate that varies by organism and environmental condition. Advances in unbiased interrogation of error types and experiments involving wild organisms may help our understanding of the potentially adaptive roles for protein translation errors. © 2014 Elsevier Ltd.


Toosky M.,Tsinghua University | Javid B.,Tsinghua University | Javid B.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | Javid B.,University of Cambridge
British Medical Bulletin | Year: 2014

Introduction Drug-resistant tuberculosis (DR-TB) is associated with increased mortality and morbidity. This is at least partly due to late diagnosis and ineffective treatment of drug-resistant status. Sources of data Selective search of the literature on DR-TB supplemented by recent guidelines from the World Health Organization. Areas of agreement Better and more rapid diagnosis of DR-TB by new techniques such as Xpert Mtb/RIF are likely to make a substantial impact on the disease. New therapeutics for DR-TB are entering, or about to enter the market for the first time in decades. Areas of controversy It is not clear whether new treatments should be restricted for DR-TB or also used for drug-susceptible tuberculosis. Growing points With several new agents on the horizon, there is the real possibility of an entirely new regimen for tuberculosis. Areas timely for developing research An inexpensive 'near-patient' diagnostic test is still needed. Optimizing new drug combination regimens in a timely manner is urgently required. © The Author 2014.


Li M.H.,Chinese National Institute for Viral Disease Control and Prevention | Li M.H.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | Fu S.H.,Chinese National Institute for Viral Disease Control and Prevention | Fu S.H.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | And 8 more authors.
Biomedical and Environmental Sciences | Year: 2014

Objective To determine the molecular characterization of full-length genome of Japanese encephalitis virus (JEV) genotype V. Methods The full-length nucleotide sequences of JEV strains isolated from different locations and sources were used in sequence and phylogenetic analysis. Results The full-length genome of genotypes V JEV, XZ0934, and Muar strain were composed of 10 983 and 10 988 nucleotides respectively and shared a lower level of identity with JEV genotypes I-IV, ranging from 78.4% (G I, KV1899) to 79.7% (G III, JaGAr01), for the nucleotide sequences, and from 90.0% (G I, KV1899) to 91.8% (G III, JaGAr01) for the amino acid sequences. The open reading frame (ORF) of JEV genotype V spanned nucleotides 96 to 10 397 and encoded 3 433 amino acids. Interestingly, a comparison with JEV genotype I-IV revealed that 3 nucleotides (encoded with a serine residue) were inserted in the NS4A gene of JEV genotype V, and the insertion of nucleotides was also found in downstream of the ORF stop codon in 3'-untranslated region. Moreover, numerous amino acid mutations were observed in 3 functional domains of the E gene of JEV genotype V. Conclusions The molecular characterization of JEV genotype V is significantly different from that of the known genotypes I-IV. The mutations located in the coding region and the non-coding region may be molecular markers of JEV genotype V and warrant further studies to determine their effects on biology and immunogenicity of genotype V strains. © 2014 The Editorial Board of Biomedical and Environmental Sciences.


Zhang W.,Chinese National Institute for Communicable Disease Control and Prevention | Zhang W.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | Du P.,Chinese National Institute for Communicable Disease Control and Prevention | Du P.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | And 6 more authors.
Journal of General and Applied Microbiology | Year: 2014

We compared pairs of 1,226 bacterial strains with whole genome sequences and calculated their average nucleotide identity (ANI) between genomes to determine whether whole genome comparison can be directly used for bacterial species definition. We found that genome comparisons of two bacterial strains from the same species (SGC) have a significantly higher ANI than those of two strains from different species (DGC), and that the ANI between the query and the reference genomes can be used to determine whether two genomes come from the same species. Bacterial species definition based on ANI with a cut-off value of 0.92 matched well (81.5%) with the current bacterial species definition. The ANI value was shown to be consistent with the standard for traditional bacterial species definition, and it could be used in bacterial taxonomy for species definition. A new bioinformatics program (ANItools) was also provided in this study for users to obtain the ANI value of any two bacterial genome pairs (http://genome.bioinfo-icdc.org/). This program can match a query strain to all bacterial genomes, and identify the highest ANI value of the strain at the species, genus and family levels respectively, providing valuable insights for species definition. © 2014 Applied Microbiology, Molecular and Cellular Biosciences Research Foundation.


Wan Q.,Zhejiang University | Zhou Z.,Zhejiang University | Ding S.,Zhejiang University | Ding S.,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease | And 2 more authors.
Journal of Interferon and Cytokine Research | Year: 2015

Interleukin-17 (IL-17) has been proved to be involved in the pathogenesis of several autoimmune diseases, including lupus, rheumatoid arthritis, multiple sclerosis, and inflammatory bowel disease. The regulation of IL-17 signal transduction is less studied. miR-30a has been identified to be downregulated in these human autoimmune diseases and their related animal models. However, how it functions in IL-17-mediated inflammation and the pathogenesis of these diseases remain unknown. In this study, we showed that miR-30a inhibits IL-17-mediated NF-κB and MAPK activation, leading to the reduced production of inflammatory cytokines and chemokines. miR-30a also reduced mRNA stability triggered by IL-17 stimulation. These suppressive effects of miR-30a were mediated by directly targeting Traf3ip2 mRNA (coding for Act1). Thus, we concluded that the downregulation of miR-30a in autoimmune diseases may exacerbate IL-17-mediated inflammation, which may serve as a potential target for the therapy of these diseases. © Copyright 2015, Mary Ann Liebert, Inc.

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