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Lee, Dae-Sik,Li, Cheng Guo,Ihm, Chunhwa,Jung, Hyungil Elsevier 2018 Sensors and actuators. B Chemical Vol.255 No.1
<P><B>Abstract</B></P> <P>In spite of the invasiveness and pain related with it, blood sampling is necessary in point-of-care systems to obtain biochemical information. In this context, a substitute micro-scale needle for minimally invasive and painless blood sampling has been developed. In conventional microneedle fabrication methods, it is difficult to combine the features of a sharp tip shape, appropriate length, and a hollow structure simultaneously. Thus, here we report a sharp, hollow, and ultrahigh aspect ratio microneedle long enough to get to the blood vessels which is capable of minimizing the pain caused by nerve pricking. It is equipped with a fluidic system to handle blood. The structure of the microneedle was precisely formed using a drawing lithography method and a sharp angle tip was formed using a laser-cutting method.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A sharp, hollow, and ultra-high-aspect-ratio microneedlelong enough to get to blood vessels and a blood-handling device are proposed. </LI> <LI> The structure of the microneedle was precisely formed using a drawing lithography and a sharp angle tip was formed using a laser-cutting method. </LI> <LI> The proposed structure shows a reasonable rigidness for penetration of skin without breaking. </LI> <LI> The microneedle with a height of 1800μm, an inner radius of 30μm, an outer radius of 50μm, and a bevel angle of 60°, works well for whole blood sampling. </LI> </UL> </P>
Development of a Novel Diagnostic Biomarker Set for Rheumatoid Arthritis Using a Proteomics Approach
Mun, Sora,Lee, Jiyeong,Lim, Mi-Kyoung,Lee, You-Rim,Ihm, Chunhwa,Lee, Seung Hoon,Kang, Hee-Gyoo Hindawi 2018 BioMed research international Vol.2018 No.-
<P><B>Background</B></P><P> Rheumatoid arthritis (RA) is an autoimmune disease that starts with inflammation of the synovial membrane. Studies have been conducted to develop methods for efficient diagnosis of RA and to identify the mechanisms underlying RA development. Blood samples can be useful for detecting disturbance of homeostasis in patients with RA. Nanoliquid chromatography-tandem mass spectrometry (LC-MS/MS) is an efficient proteomics approach to analyze blood sample and quantify serum proteins.</P><P><B> Methods</B></P><P> Serum samples of 18 healthy controls and 18 patients with RA were analyzed by LC-MS/MS. Selected candidate biomarkers were validated by enzyme-linked immunosorbent assay (ELISA) using sera from 43 healthy controls and 44 patients with RA.</P><P><B> Results</B></P><P> Thirty-eight proteins were significantly differentially expressed by more than 2-fold in healthy controls and patients with RA. Based on a literature survey, we selected six candidate RA biomarkers. ELISA was used to evaluate whether these proteins effectively allow distinguishing patients with RA from healthy controls and monitoring drug efficacy. SAA4, gelsolin, and vitamin D-binding protein were validated as potential biomarkers of RA for screening and drug efficacy monitoring of RA.</P><P><B> Conclusions</B></P><P> We identified a panel of three biomarkers for RA which has potential for application in RA diagnosis and drug efficacy monitoring. Further, our findings will aid in understanding the pathogenesis of RA.</P>
Estimation of Age of Bloodstains by Mass-Spectrometry: A Metabolomic Approach
Seok, Ae Eun,Lee, Jiyeong,Lee, You-Rim,Lee, Yoo-Jin,Kim, Hyo-Jin,Ihm, Chunhwa,Sung, Ho Joong,Hyun, Sung Hee,Kang, Hee-Gyoo American Chemical Society 2018 ANALYTICAL CHEMISTRY - Vol.90 No.21
<P>Bloodstains are common evidence in crime scenes, containing significant information, including genetic information. Although efforts have been made to reliably determine the time of incident by analyzing the elapsed time of the bloodstain, there has been limited success. To identify candidate metabolites in bloodstains over time, we prepared bloodstain samples using filter paper and analyzed the metabolites by high-performance liquid chromatography-mass spectrometry (HPLC-MS)/MS over a 21-day period. Using Venn diagrams and by multivariate analysis, we selected 62 candidate molecular features. We found by partial least-squares discriminant analysis (PLS-DA) that the group can be classified with an accuracy of 75.0%, and the <I>R</I><SUP>2</SUP> and <I>Q</I><SUP>2</SUP> values were 0.7513 and 0.6998, respectively. Five metabolites were successfully identified based on candidate molecular features. The level of two metabolites, <SMALL>L</SMALL>-tryptophan and ergothioneine, decreased with time. The concentration of candidate metabolites that we propose reliably increased or decreased with time, thus, enabling the measurement of elapsed time of the bloodstain. This study is the first to identify markers used to analyze the elapsed time of bloodstains through metabolomics analysis.</P> [FIG OMISSION]</BR>
Ju, Hyoungseok,Lim, Byungho,Kim, Minjin,Noh, Seung‐,Moo,Kim, Woo Ho,Ihm, Chunhwa,Choi, Bo Youl,Kim, Yong Sung,Kang, Changwon Wiley Subscription Services, Inc., A Wiley Company 2010 Cancer Vol.116 No.18
<P><B>Abstract</B></P><P><B>BACKGROUND:</B></P><P>A primary inhibitor of plasminogen activators, SERPINE1 (serpin peptidase inhibitor 1, clade E, member 1, also known as plasminogen activator inhibitor type 1), is an important regulator in tumorigenesis and is highly expressed in many cancers.</P><P><B>METHODS:</B></P><P>Five tag single nucleotide polymorphisms (SNPs) and 1 insertion polymorphism within <I>SERPINE1</I> were genotyped in 1101 unrelated Korean individuals (a case group of 612 patients with gastric cancer and a control group of 489 healthy individuals). Associations with susceptibility to diffuse‐type gastric cancer (DGC) and intestinal‐type gastric cancer were assessed using multivariate logistic regression analyses adjusted for age and sex. Allelic differences in the contribution to gene expression were measured using luciferase assays.</P><P><B>RESULTS:</B></P><P>SNP c.1162+162C>T (rs2227692) in intron 7 was associated with susceptibility to DGC but not with susceptibility to intestinal‐type gastric cancer. The minor allele‐carrying genotypes C/T and T/T had 1.6‐fold greater odds of DGC than the C/C genotype (<I>P</I> = .00084). This SNP was linked to a repeat‐number variation c.1162+604AAAG(11_17), a deletion (del) variation c.1162+664_1162+673del, and another SNP c.1162+859T>A (rs2070683) in intron 7 based on the sequencing of 5 patients and 5 controls. The risk haplotype of the 4 variations exhibited a 30% greater gene expression level than the nonrisk haplotype in luciferase reporter assays (<I>P</I> = .025). In contrast, DGC susceptibility was not associated with the c.−1969_−1968insG polymorphism (rs1799768) in the promoter, commonly known as 4G/5G, in which the minor 5G allele is less active in transcription than the major 4G allele.</P><P><B>CONCLUSIONS:</B></P><P>An association between <I>SERPINE1</I> and DGC susceptibility was observed with 4 correlated polymorphisms in intron 7 rather than the 4G/5G polymorphism in the promoter, although all polymorphisms affected gene expression. Cancer 2010. © 2010 American Cancer Society.</P>
Lee, You-Rim,Lee, Jiyeong,Seok, Ae Eun,Kim, Hyo-Jin,Lee, Yoo-Jin,Ihm, Chunhwa,Sung, Ho Joong,Hyun, Sung Hee,Kang, Hee-Gyoo Elsevier 2019 Forensic science international Vol.294 No.-
<P><B>Abstract</B></P> <P>Analysis of the components of bloodstains found at crime scenes can provide important information for solving the crime. However, components of blood and bloodstains vary with volume and various other unpredictable factors. Therefore, it is necessary to specify the volume of the initial liquid blood droplet and standardize the analysis. In this study, internal standard metabolites that remained constant in a certain amount of bloodstain, long after deposition of the stain, were identified. Liquid chromatography–electrospray ionization–tandem mass spectrometry of the metabolites extracted from the bloodstain samples at various time points (0, 7, 14, 21, and 28 days) was performed. The coefficient of variation (CV) of the obtained molecular features was calculated for each criterion: time point, subject, and all data (time and subject, triplicate of each). Five molecular features with average CVs of less than or equal to 5% were selected as candidates. Partial least squares discriminant analysis and principal component analysis showed that the effect on the candidates was very low over time. The fold-change value of abundances was confirmed according to time. Stigmasterol exhibited the most stable pattern; <SMALL>L</SMALL>-methionine remained stable until day 14 and after day 21. This study was the first attempt to identify internal standard metabolites that were maintained at a constant level in a bloodstain for a sufficiently long time. Analysis of internal standard metabolites in bloodstains will facilitate determination of the initial blood volume from which the bloodstain was made. Moreover, this method will provide an approach for standardization of bloodstains to obtain absolute quantitative information of bloodstain components at crime scenes.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Internal standard metabolites of bloodstains were identified. </LI> <LI> Five molecular features having average CVs of≤5% were selected as candidates. </LI> <LI> PLS-DA and PCA showed that the effect on the candidates was very low over time. </LI> <LI> Stigmasterol exhibited the most stable pattern. </LI> <LI> <SMALL>L</SMALL>-methionine remained stable until day 14 and after day 21. </LI> </UL> </P>