Park, Jong-Lyul;Kim, Mirang;Song, Kyu-Sang;Kim, Seon-Young;Kim, Yong Sung
Genomics & Informatics
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v.13
no.3
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pp.70-75
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2015
MicroRNAs (miRNAs) have been demonstrated to play an important role in carcinogenesis. Previous studies revealed that miRNAs are present in human plasma in a remarkably stable form that is protected from endogenous RNase activity. In this study, we measured the plasma expression levels of three miRNAs (miR-21, miR-27a, and miR-155) to investigate the usefulness of miRNAs for gastric cancer detection. We initially examined plasma miRNA expression levels in a screening cohort consisting of 15 patients with gastric cancer and 15 healthy controls from Korean population, using TaqMan quantitative real-time polymerase chain reaction. We observed that the expression level of miR-27a was significantly higher in patients with gastric cancer than in healthy controls, whereas the miR-21 and miR-155a expression levels were not significantly higher in the patients with gastric cancer. Therefore, we further validated the miR-27a expression level in 73 paired gastric cancer tissues and in a validation plasma cohort from 35 patients with gastric cancer and 35 healthy controls. In both the gastric cancer tissues and the validation plasma cohort, the miR-27a expression levels were significantly higher in patients with gastric cancer. Receiver-operator characteristic (ROC) analysis of the validation cohort, revealed an area under the ROC curve value of 0.70 with 75% sensitivity and 56% specificity in discriminating gastric cancer. Thus, the miR-27a expression level in plasma could be a useful biomarker for the diagnosis and/or prognosis of gastric cancer.
Proceedings of the Korean Vacuum Society Conference
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2013.08a
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pp.88-89
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2013
A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.
Cai, Er-Hui;Gao, Yong-Xin;Wei, Zhong-Zhi;Chen, Wei-Ying;Yu, Ping;Li, Ke
Asian Pacific Journal of Cancer Prevention
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v.13
no.4
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pp.1563-1567
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2012
To investigate the relationship between serum miRNA-21 (miR-21) expression in esophageal squamous cell carcinomas (ESCCs) and its clinicopathologic features, a 1:1 matched case-control study including 21 patients with ESCC and 21 age- and gender-matched healthy controls was carried out. Serum specimens were taken from all subjects. Total RNA was extracted and the stem-loop real time polymerase chain reaction was used to measure serum miR-21 in both groups. Clinical parameters were assessed to determine associations with serum miR-21 concentrations. Serum miR-21 expression in ESCC samples was significantly higher than in paired cancer-free samples (P<0.05). Metastasis was associated with mir-21 expression in serum (P<0.05), ESCC patients with metastasis having 8.4-fold higher serum miR-21 concentrations than healthy controls. There were no statistically significant associations between miR-21 expression and clinicopathologic parameters, such as gender (P>0.05), age (P>0.05), tumor location (P>0.05), cell differentiation (P>0.05), TNM staging (P>0.05), whether chemo/radiotherapy had been administered (P>0.05), or whether surgery had been performed (P>0.05). These findings suggest that the detection of microRNA-21 in serum might serve as a new tumor biomarker in diagnosis and assessment of prognosis of ESCCs.
One of the most significant features of diagnostic ultrasonic instruments is to provide real time information of the soft tissues movements. Echocardiogram has been widely used for diagnosis of heart diseases since it is able to show real time images of heart valves and walls. However, the currently used ultrasonic images are deteriorated due to presence of speckle noises and image dropout. Therefore, it is very important to develop a new technique which can enhance ultrasonic images. In this study, a technique which extracts enhanced binary images in echocardiograms was proposed. For this purpose, a digital moving image file was made from analog echocardiogram, then it was stored as 8-bit gray-level for each frame. For an efficient image processing, the region containing the heat septum and tricuspid valve was selected as the region of interest(ROI). Image enhancement filters and morphology filters were used to reduce speckle noises in the images. The proposed procedure in this paper resulted in binary images with enhanced contour compared to those form the conventional threshold technique and original image processing technique which can be further implemented for the quantitative analysis of the left ventricular wall motion in echocardiogram by easy detection of the heart wall contours.
Journal of the Institute of Electronics and Information Engineers
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v.51
no.6
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pp.50-59
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2014
In mobile healthcare service, the accurate detection and the notification of the emergency situation are important to chronic patients' life. In the existing healthcare service, the medical staff or medical service provider always judges patients' health status by monitoring from the measured from bio-data. However, it is difficult to monitor many patients in real-time simultaneously, because the medical staff should monitor the health status continuously. Furthermore, an emergency condition diagnosis based solely on the statistical level of the bio-data may be difficult, since the emergency judgment of the bio-data might differ depending on the health characteristics of each person such as age, history of disease, gender, etc. In order to solve this problem, this article presents an mobile healthcare system for emergency bio-data management using a personalized emergency policy. The salient feature of the proposed mobile healthcare system is that the characteristics of the health status of an unique patient is defined to the policy, which is used to judge the emergency condition of the bio-data measured from the patient. The prototype of proposed mobile healthcare system has been built to demonstrate the design concept.
Owing to the risk of fetal loss associated with prenatal diagnostic procedures (amniocentesis, chorionic villus sampling), noninvasive prenatal diagnosis (NIPD) is ultimate goal of prenatal diagnosis. The discovery of circulating cell-free fetal DNA (cffDNA) in maternal plasma in 1997 has opened up new probabilities for NIPD by Dr. Lo et al. The last decade has seen great development in NIPD. Fetal sex and fetal RhD status determination by cffDNA analysis is already in clinical use in certain countries. For routine use, this test is limited by the amount of cell-free maternal DNA in blood sample, the lack of universal fetal markers, and appropriate reference materials. To improve the accuracy of detection of fetal specific sequences in maternal plasma, internal positive controls to confirm to presence of fetal DNA should be analyzed. We have developed strategies for noninvasive determination of fetal gender, and fetal RhD genotyping using cffDNA in maternal plasma, using real-time quantitative polymerase chain reaction (RT-PCR) including RASSF1A epigenetic fetal DNA marker (gender-independent) as internal positive controls, which is to be first successful study of this kind in Korea. In our study, accurate detection of fetal gender through gestational age, and fetal RhD genotyping in RhD-negative pregnant women was achieved. In this assay, we show that the assay is sensitive, easy, fast, and reliable. These developments improve the reliability of the applications of circulating fetal DNA when used in clinical practice to manage sex-linked disorders (e.g., hemophilia, Duchenne muscular dystrophy), congenital adrenal hyperplasia (CAH), RhD incompatibility, and the other noninvasive pregnant diagnostic tests on the coming soon. The study was the first successful case in Korea using cffDNA in maternal plasma, which has created a new avenue for clinical applications of NIPD.
Journal of the Korea Academia-Industrial cooperation Society
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v.17
no.9
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pp.292-301
/
2016
Because damages arising from the occurrence of foot-and-mouth disease (FMD) are very great, it is essential to make a preemptive diagnosis to cope with it in order to minimize those damages. The main symptoms of foot-and-mouth disease are body temperature increase, loss of appetite, formation of blisters in the mouth, on hooves and breasts, etc. in a cow or a bull, among which the body temperature check is the easiest and quickest way to detect the disease. In this paper, an algorithm to detect FMD from the hooves of cattle was developed and implemented for preemptive coping with foot-and-mouth disease, and a hoof check test is conducted after the installation of a high-resolution camera module, a thermo-graphic camera, and a temperature/humidity module in the cattle shed. Through the algorithm and system developed in this study, it is possible to cope with an early-stage situation in which cattle are suspected as suffering from foot-and-mouth disease, creating an optimized growth environment for cattle. In particular, in this study, the system to cope with FMD does not use a portable thermo-graphic camera, but a fixed camera attached to the cattle shed. It does not need additional personnel, has a function to measure the temperature of cattle hooves automatically through an image algorithm, and includes an automated alarm for a smart phone. This system enables the prediction of a possible occurrence of foot-and-mouth disease on a real-time basis, and also enables initial-stage disinfection to be performed to cope with the disease without needing extra personnel.
The Journal of the Institute of Internet, Broadcasting and Communication
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v.20
no.1
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pp.163-169
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2020
Recently, research on automation and unmanned operation of machines in the industrial field has been conducted with the advent of AI, Big data, and the IoT, which are the core technologies of the Fourth Industrial Revolution. The machines for these automation processes are controlled based on the data collected from the sensors attached to them, and further, the processes are managed. Conventionally, the abnormalities of sensors are periodically checked and managed. However, due to various environmental factors and situations in the industrial field, there are cases where the inspection due to the failure is not missed or failures are not detected to prevent damage due to sensor failure. In addition, even if a failure occurs, it is not immediately detected, which worsens the process loss. Therefore, in order to prevent damage caused by such a sudden sensor failure, it is necessary to identify the failure of the sensor in an embedded system in real-time and to diagnose the failure and determine the type for a quick response. In this paper, a deep neural network-based fault diagnosis system is designed and implemented using Raspberry Pi to classify typical sensor fault types such as erratic fault, hard-over fault, spike fault, and stuck fault. In order to diagnose sensor failure, the network is constructed using Google's proposed Inverted residual block structure of MobilieNetV2. The proposed scheme reduces memory usage and improves the performance of the conventional CNN technique to classify sensor faults.
Background: Ventilator-associated pneumonia (VAP) requires prompt and appropriate treatment. Since methicillin-resistant Staphylococcus aureus (MRSA) is a frequent pathogen in VAP, rapid identification of it, is pivotal. Our aim was to evaluate the utility of quantitative polymerase chain reaction (qPCR) as a useful method for etiologic diagnoses of MRSA pneumonia. Methods: We performed qPCR for mecA, S. aureus-specific femA-SA, and S. epidermidis-specific femA-SE genes from bronchoalveolar lavage or bronchial washing samples obtained from clinically-suspected VAP. Molecular identification of MRSA was based on the presence of the mecA and femA-SA gene, with the absence of the femA-SE gene. To compensate for the experimental and clinical conditions, we spiked an internal control in the course of DNA extraction. We estimated number of colony-forming units per mL (CFU/mL) of MRSA samples through a standard curve of a serially-diluted reference MRSA strain. We compared the threshold cycle (Ct) value with the microbiologic results of MRSA. Results: We obtained the mecA gene standard curve, which showed the detection limit of the mecA gene to be 100 fg, which corresponds to a copy number of 30. We chose cut-off Ct values of 27.94 (equivalent to $1{\times}10^4$ CFU/mL) and 21.78 (equivalent to $1{\times}10^5$ CFU/mL). The sensitivity and specificity of our assay were 88.9% and 88.9% respectively, when compared with quantitative cultures. Conclusion: Our results were valuable for diagnosing and identifying pathogens involved in VAP. We believe our modified qPCR is an appropriate tool for the rapid diagnosis of clinical pathogens regarding patients in the intensive care unit.
Breast cancer is the most prevalent type of cancer among women around the world, and mortality is primarily caused by micro-metastatic disease. The complex mechanisms of breast cancer invasion and metastasis are intrinsically related to the malignant cell type so that early detection of micro-metastases can help prolongation of survival for patient. The aim of the present research work was evaluation of the expression status of mammoglobin protein as a candidate molecular marker in the negative sentinel lymph node (SLN). Fifty tumor specimens, and 50 normal adjacent breast tissue samples from the same patients were selected on the basis of having more than 10% tumor content for RNA extraction from SLNs. Tumor samples and normal adjacent breast tissue were archived in the form of frozen fresh tissue in liquid nitrogen. Real-time PCR was performed on a Bioner life express gradient thermal cycler system. Mammoglobin gene overexpression in breast cancer metastasis was investigated. Single marker results were mammaglobin 66.7% and CK19 50.0%, with 58.3% for the two in combination. Due to improved outcome with at least 3 genes (83.3%), it seems, triple marker evaluation will be most likely useful for detecting micro-metastases instead of studying separate genes.
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