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Identification of novel biomarkers for adult-onset-immunodeficiency(AOID)syndrome using serum proteomics

Asian Pacific Journal of Tropical Medicine 2017年第5期10卷 445-452页

作者： jeerang wongtrakul Thananya Thongtan Sittiruk Roytrakul Jutarat Praparattanapan Jiraprapa Wipasa Benjawan Kumrapich Khuanchai Supparatpinyo Research Institute for Health Sciences Chiang Mai University110 Intavaroros RoadSriphumMuang DistrictChiang Mai 50200Thailand Department of Biochemistry Faculty of MedicineChulalongkorn University1873 Rama 4 RoadPathumwanBangkok 10330Thailand Proteomics Research Laboratory Genome InstituteNational Center for Genetic Engineering and Biotechnology113 Thailand Science ParkPhahonyothin RoadKhlong NuengKhlong LuangPathumthani 12120Thailand Department of Medicine Faculty of MedicineChiang Mai University110 Intavaroros RoadSriphumMuang DistrictChiang Mai 50200Thailand

Objective:To identify the candidate protein biomarkers of adult-onset-immunodeficiency(AOID) syndrome using serum proteomics. Methods:Screening and verification phases were performed in the study. A total of 97 serum samples were classified into three groups:AOID patients with opportunistic infections(active AOID),AOID patients without opportunistic infections(inactive AOID),and healthy control. In the screening phase,pooled sera collected from patients and healthy control in each group were separated by 2D-gel electrophoresis,analyzed for differentially expressed proteins and identified for biomarkers using LC/MS. In the verification phase,the protein candidates were selected for confirmation by western blotting. Results:The analysis revealed 35 differentially expressed proteins. Three proteins including haptoglobin,gelsolin,and transthyretin,were selected for verification. The results showed that the levels of haptoglobin in both active and inactive AOID groups were significantly higher than that in the control group,while the levels of gelsolin in the active AOID group were significantly lower than that in the inactive AOID group. The level of transthyretin in the active AOID group was also significantly lower than that in the control group. Conclusions:The comparison of serum proteins between the three groups revealed three candidates which are related to chronic inflammatory diseases. Haptoglobin and transthyretin biomarkers could be applied in clinical assessment for monitor of disease outcome,including for the study of AOID pathogenesis.

关键词： Adult-onset immunodeficiency syndrome Anti-interferon-gamma autoantibody Haptoglobin Gelsolin Transthyretin Biomarkers

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Nevirapine induces apoptosis in liver(HepG2) cells

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Asian Pacific Journal of Tropical Medicine 2016年第6期9卷 531-536页

作者： jeerang wongtrakul Atchara Paemanee Phitchayapak Wintachai Chutima Thepparit Sittiruk Roytrakul Thananya Thongtan Kanokwan Janphen Khuanchai Supparatpinyo Duncan R.Smith Research Institute for Health Sciences Chiang Mai University Molecular Pathology Laboratory Institute of Molecular Biosciences Mahidol University Proteomics Research Laboratory Genome Institute National Center for Genetic Engineering and Biotechnology Department of Biochemistry Faculty of Medicine Chulalongkorn University Department of Medicine Faculty of Medicine Chiang Mai University

Objective: To generate insights into the mechanism of NVP induced hepatotoxicity. Methods: Liver(HepG2) cells were cultured with various concentrations of NVP. This cell line was chosen because it has low expression of cytochrome P450, allowing evaluation of the effects of NVP rather than specific metabolites. Cytotoxicity was determined using a proliferation assay and cell numbers were monitored using trypan blue exclusion assay for long term culture experiments and apoptosis induction was determined by morphological and biochemical investigation. Results: HepG2 cells treated with the highest concentration of NVP tested(819 μM) initially showed a rounded morphology and all cells had died by week three of exposure. Nuclear condensation and fragmentation, increased Annexin V/propidium iodide staining and caspase 9 activation all supported the induction of apoptosis in HepG2 cells in response to NVP treatment. Conclusions: There is a clear induction of apoptosis in response to NVP which suggests that NVP has significant cytotoxicity, over and above any cytotoxicity of metabolites and may contribute directly to patient hepatotoxicity.

关键词： Nevirapine Apoptosis Hepatotoxicity Cytotoxicity

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