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Chen Q.,Jiangsu Province Blood Center | Chen Q.,Soochow University of China | Srivastava K.,U.S. National Institutes of Health | Liu Z.,Peking Union Medical College | And 6 more authors.
Transfusion | Year: 2016

BACKGROUND Human neutrophil antigen-3 (HNA-3) alloantibodies can cause fatal transfusion-related acute lung injury (TRALI). Most frequencies of SLC44A2 alleles encoding the HNA-3a/b antigens have been established in Han individuals by polymerase chain reaction with sequence-specific priming (PCR-SSP). We sequenced SLC44A2 gene fragments and determined allele frequencies in three ethnicities of China. STUDY DESIGN AND METHODS Genomic DNA was extracted from 448 samples of 100 blood donors of Yi ethnicity in Xichang, Liangshan; 248 Han in Nanjing, Jiangsu; and 100 Tibetan in Lhasa, Tibet. A PCR-SSP was applied to determine the phase of two single-nucleotide polymorphisms (SNPs); SLC44A2 haplotypes were constructed. RESULTS In the 567 nucleotides of the SLC44A2 gene covered by our sequencing approach in Han individuals, we detected the known 331-44G>A (rs12972963) and 461G>A (rs2288904) polymorphisms. In the 243 nucleotides sequenced in Yi and Tibetan populations, we detected the known 461G>A and 503-15T>C (rs1560711) polymorphisms. A PCR-SSP for the common HNA-3a/b SNP was 100% concordant. The frequencies of the HNA-3a allele were 0.58, 0.66, and 0.69 in Yi, Han (Nanjing), and Tibetan, respectively (0.42, 0.34, and 0.31 for HNA-3b). CONCLUSIONS The Yi population of China had the highest frequency of blood donors at risk of harboring anti-HNA-3a compared to any population studied so far. We confirmed that the underlying SLC44A2∗2 allele is more common in China than in any European or African populations. © 2015 AABB. Source

Zeng R.,Peking Union Medical College | Zeng R.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | Li M.,Peking Union Medical College | Li M.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | And 13 more authors.
Antimicrobial Agents and Chemotherapy | Year: 2014

Aspergillus fumigatus biofilms still present a challenge for effective treatment in clinical settings. While mild heat stress has been introduced as a treatment for infectious diseases, the effectiveness of mild heat stress on A. fumigatus biofilm formation and antifungal susceptibility is still unknown. In the present study, confocal laser scanning microscopy (CLSM) was used to image and quantify Aspergillus fumigatus biofilm formation under three different regimens of continuous mild heat stress: at 37, 39, and 41°C. Furthermore, fungal growth has been investigated under the above conditions in combination with antifungal drugs (amphotericin B [AMB], micafungin [MCF], and voriconazole [VOC]) at early and late stages. CLSM analysis showed that higher temperatures induce earlier germination and greater hyphal elongation but poorer polar growth and reduced biofilm thickness. In the early stage of biofilm formation, the combination of treatment at 39 or 41°C with MCF or VOC produced no visible difference in biomass formation from similar treatments at 37°C with the same drug. Interestingly, AMB treatment at 37°C inhibited early stage biofilm formation to a much greater extent than at 39 and 41°C. At the late stage of biofilm formation, the mild heat treatments at 39 and 41°C with AMB, MCF, and VOC inhibited biomass formation compared to that at 37°C. The present data show that mild heat stress has a negative regulatory effect on biofilm formation in vitro, and antifungal drug improvement with mild heat treatment at late-stage biofilm formation provides useful indications of possible effective strategies for clinical management of aspergillosis. Copyright © 2014, American Society for Microbiology. All Rights Reserved. Source

Li X.,Peking Union Medical College | Li X.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | Cai Q.,Peking Union Medical College | Cai Q.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | And 12 more authors.
Journal of Antimicrobial Chemotherapy | Year: 2014

Objectives: The histone deacetylase (HDAC) has recently been linked to the morphogenesis and virulence of yeast. However, the effects of HDAC on antifungal susceptibility are not well understood. We sought to characterize the action of histone deacetylation on azole resistance in Candida albicans and its possible mechanism of action. Methods: A total of 40 C. albicans strains were studied. Azole susceptibility with or without trichostatin A (TSA) was determined according to the CLSI microdilution method. The null mutants of HDA1 and RPD3 (genes targeted by TSA) were also investigated using drop-plate assays and a rapid acquisition of adaptation to the azole test. Transcriptional levels of HDAC genes and efflux genes were quantified using RT-PCR for both the basal and fluconazole-induced conditions. Results: The inhibition of HDACs by TSA (0.25 mg/L) markedly reduced the trailing growth and the growth of most C. albicans strains. Trailing growth for C. albicans strains was decreased from 2-fold to 256-fold at 48 h. The deletion of HDA1 or RPD3 increased the susceptibility to azoles compared with the WT strain. The expression of HDA1 and RPD3 was up-regulated to different levels, and returned to the level of the susceptible parental strain when stable resistance had formed during the course of acquired fluconazole resistance both in vitro and in vivo. Efflux genes were poorly expressed in mutant strains compared with those of the WT strain. Conclusions: Our results indicate the important role of the Rpd3/Hda1 family in the development of azole resistance in C. albicans. Histone deacetylation may govern the expression of genes related to the early stages of adaptation to azole stress, such as efflux pump genes. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. Source

Wang C.,Peking Union Medical College | Wang C.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | Zhan P.,Peking Union Medical College | Zhan P.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | And 12 more authors.
Mycopathologia | Year: 2014

Invasive aspergillosis (IA) is a major concern in patients with severe immune deficiency. As antifungal susceptibility varies in different fungal pathogens, accurate and timely identification of species is becoming imperative for guidance of therapy and reducing high mortality rates in patients with IA. But, in fact, the diagnosis is challenging and new validated techniques are required for the detection and identification of clinically relevant isolates. The laser capture microdissection (LCM) system enables analysis of cytologically and/or phenotypically defined cell types from heterogeneous tissue and has been used in diagnosis and fungal species identification in pulmonary aspergillosis of white storks. To establish the experimental foundation for clinical application of the system, we microdissected and collected Blankophor-stained single hyphal strands from tissue cryosections of murine model of invasive pulmonary aspergillosis (IPA) with A. fumigatus by LCM, subsequently processed for DNA extraction, PCR sequencing, and species molecular identification. The sensitivity of LCM-PCR sequencing was 89 % (89/100), and the specificity was 100 %. Moreover, the positive predictive value and negative predictive value were 100 and 78.43 %, respectively. The result approved that the LCM-based methods had the potential for accurately diagnosis and rapidly identification fungal pathogens of IPA. © 2014 Springer Science+Business Media Dordrecht. Source

Zhang X.,Peking Union Medical College | Zhang X.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | Liu W.,Peking Union Medical College | Liu W.,Jiangsu Key Laboratory of Molecular Biology for skin Diseases and sTIs | And 7 more authors.
Acta Dermato-Venereologica | Year: 2015

Skin and soft tissue infections caused by rapidly growing non-tuberculous mycobacteria (RG-NTM) have become a growing clinical concern over the past decades. These RG-NTM are ubiquitous environmental organisms and most are resistant to traditional antituberculous agents. In this report, we describe 3 cutaneous infections caused by RG-NTM, namely, Mycobacterium abscessus, M. chelonae, and M. conceptionense, and present the clinical and laboratory characteristics of these infections. © 2015 The Authors. Source

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