• 한국동물위생학회지
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• 제37권4호
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• pp.253-261
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• 2014

Porcine circovirus disease (PCVD) is a major problem of swine industry worldwide, and diagnosis of PCV2, causal agent of PCVD, has been doing in clinical laboratories of pig disease by polymerase chain reaction (PCR) methods. But the PCR analyses have a serious problem of misdiagnosis by contamination of DNA, in particular, from carryover contamination with previously amplified DNA or extracted DNA from field samples. In this study, an uracil DNA glycosylase (UNG)-based direct PCR (udPCR) without DNA extraction process and DNA carryover contamination was developed and evaluated on PCV2 culture and field pig samples. The sensitivity of the udPCR combined with dPCR and uPCR was same or better than that of the commercial PCR (cPCR) kit (Median diagnostics, Korea) on PCV2-positive serum, lymph node and lung samples of the pigs. In addition, the udPCR method confirmed to have a preventing ability of mis-amplification by contamination of pre-amplified PCV2 DNA from previous udPCR. In clinical application, 170 pig samples (86 tissues and 84 serum) were analysed by cPCR kit and resulted in 37% (63/170) of positive reaction, while the udPCR was able to detect the PCV2 DNA in 45.3% (77/170) with higher sensitivity than cPCR. In conclusion, the udPCR developed in the study is a time, labor and cost saving method for the detection of PCV2 and providing a preventing effect for DNA carryover contamination that can occurred in PCR process. Therefore, the udPCR assay could be an useful alternative method for the diagnosis of PCV2 in the swine disease diagnostic laboratories.

• 대한치과기공학회지
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• 제33권1호
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• pp.25-35
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• 2011

Purpose: There are some advantages of the acrylic resin denture base ; appropriate strength, volume safety, simple processing apparatus, and low cost. But, it have a weakness for fracture by intense pressure or shock. However, the repairs for resin denture base are possible using various materials and techniques. There is a few studies in repairs for resin denture base, but not clinical researches. And there is no studies in absorbed saliva into the region of fracture and bond strength. This study is to observe re-bond strength of resin denture base after repairing under saliva absorption. Methods: The samples were made of heat curing resin and the rectangular parallelepiped specimens which were 50mm long, 10mm wide and 3mm high. The four different groups immersed in the artificial saliva for 2 weeks were prepared, 1) no repaired control samples, 2) immediately repaired samples, 3) repaired samples after 1 day dry, and 4) repaired samples after 3 days dry. The prepared samples were repaired by two different curing materials, self curing resin and heat curing resin method. Each groups composed of 10 specimens were experimented with the three point bending tests for bonding strength measuring Results: There were under condition absorbed in the artificial saliva and repaired by self curing resin method, repaired specimens after 1 day and 3 days dry groups had higher values of bonding strengths than control group, and bonding strengths of immediately repaired samples were similar to those of control samples (p<0.05). There were under condition immersed in the artificial saliva and repaired by heat curing resin method, immediately repaired samples showed similar values to bonding strengths of control groups, and repaired samples after 1 day and 3 days dry groups were lower than those of control group (p>0.05). Conclusion: In this study, the repairs for resin denture base were remarkably high values of bonding strengths than those of the past, and showed that have stable bonding strengths independent of saliva absorption of denture base, so present repairs for resin denture base can be performed, regardless of saliva conditions.

• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7575-7581
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• 2014

MicroRNAs (miRNAs) play an essential role in the development and progression of nasopharyngeal carcinomas (NPC). Despite advances in the field of cancer molecular biology and biomarker discovery, the development of clinically validated biomarkers for primary NPC has remained elusive. In this study, we investigated the expression and clinical significance of miRNAs as novel primary NPC diagnostic biomarkers. We used an array containing 2, 500 miRNAs to identify 22 significant miRNAs, and these candidate miRNAs were validated using 67 fresh NPC and 25 normal control tissues via quantitative real-time PCR (qRT-PCR). Expression and correlation analyses were performed with various statistical approaches, in addition to logistic regression and receiver operating characteristic curve analyses to evaluate diagnostic efficacy. qRT-PCR revealed five differentially expressed miRNAs (miR-93-5p, miR-135b-5p, miR-205-5p and miR-183-5p) in NPC tissue samples relative to control samples (p<0.05), with miR-135b-5p and miR-205-5p being of significant diagnostic value (p<0.01). Moreover, comparison of NPC patient clinicopathologic data revealed a negative correlation between miR-93-5p and miR-183-5p expression levels and lymph node status (p<0.05). These findings display an altered expression of many miRNAs in NPC tissues, thus providing information pertinent to pathophysiological and diagnostic research. Ultimately, miR-135b-5p and miR-205-5p may be implicated as novel NPC candidate biomarkers, while miR-93-5p, miR-650 and miR-183-5p may find application as relevant clinical pathology and diagnostic candidate biomarkers.

• Translational and Clinical Pharmacology
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• 제26권4호
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• pp.160-165
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• 2018

Indobufen ($Ibustrin^{(R)}$), a reversible inhibitor of platelet aggregation, exists in two enantiomeric forms in 1:1 ratio. Here, we characterized the anti-platelet effect of S- and R-indobufen using response surface modeling using $NONMEM^{(R)}$ and predicted the therapeutic doses exerting the maximal efficacy of each enantioselective S- and R-indobufen formulation. S- and R-indobufen were added individually or together to 24 plasma samples from drug-naïve healthy subjects, generating 892 samples containing randomly selected concentrations of the drugs of 0-128 mg/L. Collagen-induced platelet aggregation in platelet-rich plasma was determined using a Chrono-log Lumi-Aggregometer. Inhibitory sigmoid $I_{max}$ model adequately described the anti-platelet effect. The S-form was more potent, whereas the R-form showed less inter-individual variation. No significant interaction was observed between the two enantiomers. The anti-platelet effect of multiple treatments with 200 mg indobufen twice daily doses was predicted in the simulation study, and the effect of S- or R-indobufen alone at various doses was predicted to define optimal dosing regimen for each enantiomer. Simulation study predicted that 200 mg twice daily administration of S-indobufen alone will produce more treatment effect than S-and R-mixture formulation. S-indobufen produced treatment effect at lower concentration than R-indobufen. However, inter-individual variation of the pharmacodynamic response was smaller in R-indobufen. The present study suggests the optimal doses of R-and S-enantioselective indobufen formulations in terms of treatment efficacy for patients with thromboembolic problems. The proposed methodology in this study can be applied to the develop novel enantio-selective drugs more efficiently.

• Tuberculosis and Respiratory Diseases
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• 제53권6호
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• pp.635-643
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• 2002

연구배경 : 완전 자동화된 액체배양법과 기존의 고체배양법을 이용하여 폐결핵환자의 객담내 mycobacterium의 신속검출율 및 양성판정시까지의 시간을 비교하고자 하였다. 방 법 : 2002년 1월 1일부터 2002년 6월 30일 사이에 본원에 입원 혹은 외래에서 폐결핵으로 치료중인 환자 127명의 객담검체를 이용하였으며, 배양 중 오염된 28개의 검체(MB/BacT system 18개, L-J배지 14개, 4검체는 두 배양법 모두에서 오염)를 제외한 99명의 객담검체를 대상으로 하였다. 검체는 4% NaOH로 전처치한 후 4300rpm으로 20분간 원심분리한 후 멸균된 phosphate buffer(pH 6.8)를 첨가하여 2ml로 만들어 MB/BacT bottle에 0.5ml, Lowenstein-Jensen배지에 0.25ml를 접종한 후 $35-37^{\circ}C$에 6-12주간 배양하면서 균검출율과 양성판정시까지의 시간을 비교하였다. 결 과 : MB/Bact system과 L-J배양볍 중 어느 한 방법에서 균성장이 발견된 검제는 677B (67.7%) 였다. 두배양법 모두에서 균성장이 발견된 겸체는 5252.5%) 개였으며, MB/BacT에서만 발견된 검체는 15개 (15.2%) 였으나 L-J배양법에서만 발견된 경우는 없었다. 객담검체의 ZN염색법으로 항산균 집균도말겸사상 양성인 검체는 58개, 음성인 검체는 41개였다. 58개의 도말양성 검체 가운데 MB/BacT system에 양성을 보인 검체는 %개 (96.6%), L-J배지양성은 467B (79.3%) 였으며, 균성장 평균 발견시간은 각각 11.0일과 23.5일이었다. 41개의 도말음성검체 가운데 MB/BacT system에 양성을 보인 검체는 11개 (26.8%), L-J배지양성은 6개 (14.6%)였으며, 균성장 평균 발견시간은 각각 13.7일과 27.6일 이었다. 전체적으로 MB/BacT system과 L-J배양법에서 균성장 발견시 간은 각각 $13.3{\pm}8.3$일, $27.2{\pm}0.9$일이었다. 결 론 : 완전 자동화된 MB/BacT system은 L-J배지에 비해 mycobacterium의 검출율이 높고 발견시간이 단축되었다. MB/BacT는 방사성 물질을 사용하지 않는 장점이 있으므로 모든 검사실에 사용하기에 적당할 것으로 보이며, 균동정을 위하여 특이한 DNA probe와 병용하면 CDC가 권고한 결과보고 시간에 근접할 수 있을 것으로 생각되었다.

• Clinical and Experimental Reproductive Medicine
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• 제33권1호
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• pp.61-61
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• 2006

목 적: 본 연구는 methylenetetrahydrofolate reductase (MTHFR C677T와 A1298C) 유전자 돌연변이형이 자연유산아 발생의 원인 유전자로 작용하는지에 대해 알아보고자 시도하였다. 연구방법: 95명의 자연유산아 조직과 대조군으로 100명의 정상 소아의 혈액 그리고 449명의 정상 성인의 혈액을 채취하여 DNA를 분리하여 사용하였다. 유전자형은 분리된 DNA를 이용하여 중합효소 연쇄반응과 제한효소 절편다형 분석방법으로 결정하였다. 결 과: 자연유산아 그룹은 소아대조군에서 보다 MTHFR 677CC 형 (p=0.014)은 높게, 677CT형 (p=0.063)은 낮게 나타났다. 성인대조군과의 비교에서도 MTHFR 677CT 형의 빈도는 현저히 낮게 나타났다 (p=0.032). 그리고 MTHFR 677CC/1298AC 조합형 유전자의 경우 소아대조군 (p=0.034)과의 비교에서는 현저히 높은 빈도를 나타냈으나, 성인대조군 (p=0.063)과의 비교에서는 높은 경향성은 있었으나 통계학적으로 유의한 차이는 없었다. 결 론: MTHFR 677CC와 MTHFR 677CC/1298AC 유전자형은 자연유산아 발생의 위험인자일 가능성이 높으며, 지속적인 연구가 요구된다.

• 대한의생명과학회지
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• 제22권2호
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• pp.37-45
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• 2016

High risk human papillomavirus (HR-HPV) is major risk factor for uterine cervical cancer. There are approximately 15 types of HR-HPV. Liquid based cytology samples (116 samples) with high grade cervical lesions belonging to cervical intraepithelial neoplasia (CIN) 2, CIN 3, carcinoma in situ (CIS) and squamous cell carcinoma (SCC) were used after histologic confirmation. HR-HPV genotype assay was conducted using DNA chips. The HR-HPV infection rate was 81.9% with SCC samples showing the highest HR-HPV infection rate of 31%. CIN 3, CIS and CIN 2 showed infection rates of 25%, 16.4% and 9.5%, respectively. According to age with HR HPV infection rate, the 30~39 years-old group showed the highest infection rate by 92.3%. According to distribution with HR HPV genotyping, HPV 16 showed the highest infection rate by 42.3% whereas HPV 33 and HPV 58 showed infection rates of 11.7% and 10.8%, respectively. HPV 18 which is the second most common infected HPV genotype in the world showed 3.6%. Of the three most common oncogenic HR-HPV genotypes in CIN 2, we detected HPV 16, 35, 58; CIN 3 was HPV 16, 33, 58; CIS was HPV 16, 58, 33 (35/52); and SCC was HPV 16, 33, and 18 (31/52/58). Among the HPV 18, CIN 2, CIN 3, CIS and SCC showed 0.9%, 0.9%, 0% and 1.8%, respectively. The most often used preventive vaccines for cervical cancers use HPV 16 and HPV 18 as targets. However, results derived from this study suggest that a preventive vaccine against HPV 16 and HPV 18 would not be optimal for populations in this study.

• Journal of Microbiology and Biotechnology
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• 제22권9호
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• pp.1301-1306
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• 2012

We combined real-time RT-PCR and real-time PCR (R/P) assays using a hydrolysis probe to detect Mycobacterium tuberculosis complex (MTBC)-specific 16S rRNA and its rRNA gene (rDNA). The assay was applied to 28 non-respiratory and 207 respiratory specimens from 218 patients. Total nucleic acids (including RNA and DNA) were extracted from samples, and results were considered positive if the repeat RT-PCR threshold cycle was ${\leq}35$ and the ratio of real-time RT-PCR and real-time PCR load was ${\geq}1.51$. The results were compared with those from existing methods, including smear, culture, and real-time PCR. Following resolution of the discrepant results between R/P assay and culture, the overall sensitivity, specificity, positive predictive values (PPV), and negative predictive values (NPV) of all samples (including non-respiratory and respiratory specimens) were 98.2%, 97.2%, 91.7%, and 99.4%, respectively, for R/P assay, and 83.9%, 89.9%, 72.3%, and 94.7%, respectively, for real-time PCR. Furthermore, the R/P assay of four patient samples showed a higher ratio before treatment than after several days of treatment. We conclude that the R/P assay is a rapid and accurate method for direct detection of MTBC, which can distinguish viable and nonviable MTBC, and thus may guide patient therapy and public health decisions.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5167-5170
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• 2012

Chilean red chili peppers contaminated with aflatoxins were reported in a previous study. If the development of gallbladder cancer (GBC) in Chile is associated with a high level of consumption of aflatoxin-contaminated red chili peppers, such peppers from other countries having a high GBC incidence rate may also be contaminated with aflatoxins. We aimed to determine whether this might be the case for red chili peppers from Bolivia and Peru. A total of 7 samples (3 from Bolivia, 4 from Peru) and 3 controls (2 from China, 1 from Japan) were evaluated. Aflatoxins were extracted with acetonitrile:water (9:1, v/v) and eluted through an immuno-affinity column. The concentrations of aflatoxins B1, B2, G1, and G2 were measured using high-performance liquid chromatography (HPLC), and then the detected aflatoxins were identified using HPLC-mass spectrometry. In some but not all of the samples from Bolivia and Peru, aflatoxin B1 or aflatoxins B1 and B2 were detected. In particular, aflatoxin B1 or total aflatoxin concentrations in a Bolivian samples were above the maximum levels for aflatoxins in spices proposed by the European Commission. Red chili peppers from Bolivia and Peru consumed by populations having high GBC incidence rates would appear to be contaminated with aflatoxins. These data suggest the possibility that a high level of consumption of aflatoxin-contaminated red chili peppers is related to the development of GBC, and the association between the two should be confirmed by a case-control study.

• Genomics & Informatics
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• 제14권1호
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• pp.34-40
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• 2016

Due to advances in omics technologies, numerous genome-wide studies on human samples have been published, and most of the omics data with the associated clinical information are available in public repositories, such as Gene Expression Omnibus and ArrayExpress. While analyzing several public datasets, we observed that errors in gender information occur quite often in public datasets. When we analyzed the gender description and the methylation patterns of gender-specific probes (glucose-6-phosphate dehydrogenase [G6PD], ephrin-B1 [EFNB1], and testis specific protein, Y-linked 2 [TSPY2]) in 5,611 samples produced using Infinium 450K HumanMethylation arrays, we found that 19 samples from 7 datasets were erroneously described. We also analyzed 1,819 samples produced using the Affymetrix U133Plus2 array using several gender-specific genes (X (inactive)-specific transcript [XIST], eukaryotic translation initiation factor 1A, Y-linked [EIF1AY], and DEAD [Asp-Glu-Ala-Asp] box polypeptide 3, Y-linked [DDDX3Y]) and found that 40 samples from 3 datasets were erroneously described. We suggest that the users of public datasets should not expect that the data are error-free and, whenever possible, that they should check the consistency of the data.