• Korean Journal of Clinical Laboratory Science
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• v.41 no.4
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• pp.145-152
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• 2009

Novel influenza A virus, subtype H1N1 of swine-lineage, has been transmitted rapidly to many regions of the world. Rapid detection of the virus is essential to instigate appropriate patient care and public health management and for disease surveillance. The aim of this study is to determine the prevalence of novel influenza A (H1N1) virus in Korea using reverse-transcription real time polymerase chain reaction (rRT-PCR). Novel H1N1 virus was detected in a total of 8,948 nasopharyngeal samples from patients with influenza-like illness throughout Korea from August to September 2009. RNA was extracted from $300{\mu}l$of sample using an RNA extraction kit (Zymo Research, CA, USA). In the present study, Genekam kit (Genekam, Duisburg, Germany) was used to detect novel H1N1 virus. Novel H1N1 virus was found in 1,130 samples from a total of 8,948 samples (12.6%). The highest frequency was found in 10- to 19-year-olds (M: 29.3% vs. F: 16.4%), followed by 20- to 29-year-olds (M: 17.9% vs. F: 15.4%), 40- to 49-year-olds (M: 6.5% vs. F: 8.1%), 50- to 59-year-olds (M: 6.0% vs. F: 5.5%), and 30- to 39-year-olds (M: 4.6% vs. F: 3.8%). The mean positive rate was higher in men than in women (M: 14.7% vs. F: 7.4%). Novel H1N1 virus showed the lowest prevalence in patients over 60 years old. The positive rate increased daily and showed a significant high peak in mid-September 2009. In 19 provinces of Korea, Cheonan (41.1%), Busan (37.3%), Gangneung (33.3%), Jinju (32.1%), Ulsan (24.6%), Deajeon (23.7%) areas showed high frequencies and other provinces were found less than 10% of novel H1N1 virus. Since reverse-transcription real time PCR assay is rapid, accurate, and convenient, it may assist public health laboratories in detecting novel H1N1 virus. Moreover, these data could be useful for the management of patients with influenza-like illness.

• Journal of Food Hygiene and Safety
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• v.25 no.4
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• pp.376-387
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• 2010

Recently, speedy, convenient and easy detection technologies have been developed rapidly and on the contrary, studies on development of traditional detectors applying biochemical characteristics has gradually been decreased. This review examined trend in current studies on detection of food-borne pathogenic microorganisms in the fields of selective media, immuno-assay, Polymerase Chain Reaction (PCR), microarray, terahertz spectroscopy & imagination and so on. Most traditional methods to detect the organisms from food matrix rely on selective media and such a method have disadvantages like long time requirement and distinguishing one species only from each selective medium although they are highly economical. Various new convenient methods such as Enzyme Linked Immuno-sorbent Assay (ELISA), paper-strip kit, fluoroimmunoassay etc. have been developed. The most ideal method for detecting food-borne pathogenic microorganisms in foods should be accurate, convenient, rapid and economical. Additionally, it is needed that capabilities of quantitative analysis and automation to be applied to industries.

• Korean Journal of Pharmacognosy
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• v.42 no.4
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• pp.348-353
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• 2011

The aim of the present study is to investigate the cytokine production inhibitory effect of a Dictamni Radicis Cortex (DRC). DRC has been commonly used as important medicinal herb in China and it used to control eczema, atopic dermatitis, fever and inflammatory diseases. Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the DRC on proinflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of DRC, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the DRC reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the DRC may have an anti-inflammatory property through suppressing inflammatory mediator productions.

• Journal of Acupuncture Research
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• v.27 no.3
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• pp.65-73
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• 2010

Objectives : Recently, Pharmacopuncture therapy has been used for the treatment of inflammatory diseases such as rheumatoid arthritis. Especially, we have been interested in chemical mediators concerned with inflammation such as prostaglandin, nitric oxide. The purpose of this study is investigated that the effect of Ampelopsis Radix Pharmacopuncture solution in RAW 264.7 macrophages, performed several experimental items : those are Prostaglandin $E_2$, Nitric Oxide and Cyclooxygenase-2. Methods : The cytotoxicity of Ampelopsis Radix Pharmacopuncture solution in RAW 264.7 macrophages were measured by MTT assay. In order to observe cyclooxygenase-2 mRNA expression in lipopolysaccharide and interferon-gamma stimulated RAW 264.7 macrophages, RT-PCR was used. Prostaglandin $E_2$ production and Nitric Oxide production was measured by nitric oxide detection kit and Prostaglandin $E_2$ assay kit. Results : 1. The MTT assay demonstrated that cytotoxic effect of Ampelopsis Radix Pharmacopuncture solution in RAW 264.7 macrophages was not appeared. 2. Ampelopsis Radix Pharmacopuncture solution inhibited nitric oxide production in lipopolysaccharide and interferon-gamma stimulated RAW 264.7 macrophages. 3. Ampelopsis Radix Pharmacopuncture solution inhibited cyclooxygenase-2 mRNA expression in lipopolysaccharide and interferon-gamma stimulated RAW 264.7 macrophages. 4. Ampelopsis Radix Pharmacopuncture solution inhibited Prostaglandin $E_2$ production in lipopolysaccharide and interferon-gamma stimulated RAW 264.7 macrophages. Conclusions : On the basis of these results, It was shown that Ampelopsis Radix Pharmacopuncture solution was able to inhibit the production of $PGE_2$ and NO, as well as COX-2 mRNA expression. Our results may provide new mechanism by which Ampelopsis Radix Pharmacopuncture solution accounts for its beneficial effect on accelerating wound healing and anti-inflammation.

• Journal of Gastric Cancer
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• v.4 no.4
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• pp.268-271
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• 2004

Purpose: Most gastrointestinal stromal tumors (GISTs) have gain-of-function mutations of the KIT or the platelet-derived growth factor receptor alpha (PDGFRA) genes, but approximately $10\%$ of the GISTs are wild types for both the KIT and the PDGFRA genes. The purpose of this study was to investigate the possibility that epidermal growth factor receptor (EGFR) gene mutation might be responsible for the pathogenesis of GIST. Materials and Methods: We analyzed the EGFR gene in 60 GISTs for the detection of somatic mutations by using the polymerase chain reaction (PCR), the single strand conformation polymorphism (SSCP), and DNA sequencing in exon 18, 19, and 21 encoding the kinase domain. Results: The SSCP analysis revealed no evidence of EGFR mutations in exon 18, 19, and 21 in GISTs. Conclusion: The data indicate that the EGFR gene may not be mutated in human GIST and suggest that therapies targeting the mutated EGFR gene products might not be useful in the treatment of GISTs.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.4
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• pp.1851-1857
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• 2014

Objective: The main purpose of this work was to investigate the effect of berberine hydrochloride (BH) on the proliferation, apoptosis, migration, and invasion of CNE-1 nasopharyngeal carcinoma cells. Our results shed light on the functional components of traditional Chinese herbs for potential use in modern medicine. Methods: The CNE-1 cell line was treated with different concentrations of BH and effects on cell viability and proliferation were evaluated using the Cell Counting Kit-8 (CCK-8) assay. Anti-migratory and anti-invasive actions of BH were investigated using wound healing assays and the Millicell Hanging cell culture insert system, respectively. Expression of the epithelial-mesenchymal transition (EMT)-related gene twist (Twist) was analyzed by real-time PCR and Western blotting. Apoptosis was estimated with an annexin-V fluorescein (FITC) apoptosis detection kit, as well as with reference to levels of activated caspase-3 of CNE-1 cells before and after treatment with BH utilizing fluorescence spectroscopy. Results: BH was capable of reducing proliferation and viability of CNE-1 cells in a dose- and time-dependent manner, also demonstrating anti-migratory and anti-invasive capacities which correlated with reduction in expression of Twist. Finally, BH was able to induce significant amounts of apoptosis in CNE-1 cells, as demonstrated by an increase in the activity of caspase-3 and in annexin-V staining following treatment. Conclusion: BH extracted from rhizoma coptidis demonstrated an ability to block proliferation, induce apoptosis, and impair the migration and invasion of the CNE-1 cell line Considering these properties, our results suggest that BH could be an important compound for consideration in the treatment of nasopharyngeal carcinoma.

• Korean Journal of Head & Neck Oncology
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• v.14 no.2
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• pp.199-205
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• 1998

Objective: Telomerase, a specialized ribonucleoprotein polymerase associated with cellular immortality, is expressed by most malignant cells and is inactive in most normal somatic cells. The assays of telomerase activity in various tumors have provided both diagnostic and prognostic information. This study was carried out to determine whether telomerase activity could be useful in distinguishing benign and malignant thyroid diseasees. Materials & Methods: Telomerase activity was determined using Oncor $TRAP_{EZE}^{TM}ELISA$ Telomerase Detection Kit for performing PCR-based telomeric repeat amplification protocol (TRAP) assay followed by ELISA detection in both normal and tumor tissues of 23 adenomatous hyperplasias, 12 follicular adenomas, 4 follicular carcinomas, 16 papillary carcinomas, 4 Hashimoto's thyroiditises and 3 malignant lymphomas. We also examined all cases microscopically to review the status of lymphoid infiltrate. Results: Of the 62 cases, extensive lymphoid infiltrates were contained in 20 tumor tissues(4 Hashimoto's thyroiditises, 3 malignant lymphomas, 6 adenomatous hyperplasias and 7 papillary carcinomas), all of which showed positive telomerase activity. All the normal tissues without lymphoid infiltrates(n=43) did not express telomerase activity. Of 42 tumor tissues without lymphoid infiltrates, 37(88.0%) showed positive telomerase activity: 13 of 17 adenomatous hyperplasias(76.5%), 11 of 12 follicular adenomas(91.7%), 4 of 4 follicular carcinomas(100.0%) and 9 of 9 papillary carcinomas(100.0%). Conclusions: Our methods showed high sensitivity in the detection of telomerase activity and the exclusion of lymphoid infiltrates may be important in telomerase assay. In our work, the measurement of telomerase activity was not useful in distinguishing benign and malignant thyroid diseases.

• The Journal of Internal Korean Medicine
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• v.33 no.4
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• pp.465-475
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• 2012

Objectives : This study was performed to investigate the antineoplastic effect of ethanol extracts from Sanguisorbae Radix on cholangiocarcinoma cells that was established from biliary tract cancer tissue. Materials and Methods : Two cholangiocarcinoma cell lines, SNU-1079 and SNU-1196, were studied. The mRNA expression of Caspase 3, 8, 9, Bcl-2, Bax, P53, and P21 was examined by RT-PCR. Cell viability was determined by MTT assay. The cell cycle was analyzed by flow cytometry and apoptosis by cell death detection ELISA kit. Results : Proliferation of SNU-1079 and SNU-1196 was inhibited by Sanguisorbae Radix treatment in a dose-dependent manner. All cells treated with Sanguisorbae Radix showed increased dose- and time-dependent apoptosis. The expression of caspase 3, 8, 9, p53, and p21 was increased in all cells after the treatment of Sanguisorbae Radix. The expression of Bcl-2 was decreased in SNU-1196 and Bax expression was increased in all cells after the treatment of Sanguisorbae Radix. Conclusions : These results suggest that Sanguisorbae Radix would be beneficial in the treatment of cholangiocarcinoma.

• Journal of Animal Science and Technology
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• v.49 no.5
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• pp.559-568
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• 2007

Recently, copy number variations (CNV) of genes or genomic segments have been intensively studied and various analysis methods have been developed. In this study, quantitative oligonucleotide ligation assay (qOLA) was applied to investigate CNV of KIT gene in the Landrace breed. A combined assay using qOLA and pyrosequencing, 6 genotype classes, I1/I1 or I3/i (IBe), I1/I2 or I3/IP, I1/I3, I1/IP or I2/i (IBe), I2/I2and I2/IP, were identified from 44 Landrace pigs. Genotype assignment using grouping features of measurements on a scatter plot showed 100% agreement with those using a statistical assignment by PROC FASTCLUS procedure implemented in the SAS package. Two versions (3100 and 3130) of ABI sequencers gave the same genotyping results, indicating there was no influence on qOLA by different versions of instrument, however, the means of standard deviation and coefficient of variation from the qOLA on a ABI 3130 (2.33 and 4.10) was lower than those from the qOLA on a ABI 3100 (2.67 and 4.81). Effect of proteinase K treatment on the PCR product followed by qOLA was very clear because noise peaks were disappeared and the observed ration fit better to the reference ratio corresponding to each genotype.

• Korean Journal of Food Science and Technology
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• v.43 no.2
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• pp.161-168
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• 2011

The purpose of this study was to evaluate microbiological contamination of fresh vegetables in Korea. Twenty types of vegetables were tested for total aerobic bacteria, coliforms, Escherichia coli, yeast and mold, and pathogenic bacteria such as Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella, E. coli O157:H7, Cronobacter sakazakii, Shigella, and Campylobacter. Levels of total aerobic bacteria and coliform on 20 vegetables were between 3.74 and 8.04 log CFU/g, and 0.16 and 5.02 log CFU/g, respectively. The highest contamination levels of total aerobic bacteria were observed on water dropwort, sprouts, mungbean sprout, and ballflower root. There was no significant difference (p>0.05) in microbial contamination levels of total aerobic count, coliform, E. coli, yeast and mold between organic and nonorganic vegetables. When isolation methods using selective agars were applied, L. monocytogenes, B. cereus, Salmonella and Campylobacter were isolated from some fresh vegetable samples. Results of API kit tests showed that L. monocytogenes was identified on Chinese cabbage, cucumber, soybean sprouts, and iceberg lettuce while Salmonella was identified on Korean leek. Furthermore, Campylobacter jejuni was also identified in more than 50 of the 100 samples. However, when positive samples from API kit were tested for real-time PCR or 16S rRNA sequencing method, only B. cereus from perilla leaf, carrot, water dropwort, and sprouts showed positive results. These results indicate that selective agar and API kit detection methods might result in false positive results for some pathogens. Therefore, studies need to improve isolation or confirmation methods for such pathogens.