• Title/Summary/Keyword: PCR (Polymerase Chain Reaction)

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A Case of Tuberculous Otitis Media in Children (소아에서 결핵성 중이염 1례)

  • Kim, Eun Ah;Ham, Se Chang;Kim, Young Kyoun;Park, Yong Won;Kang, Yun Kyung
    • Pediatric Infection and Vaccine
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    • v.9 no.1
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    • pp.110-116
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    • 2002
  • Tuberculous otitis media is a very rare disease in extrapulmonary tuberculosis and in chronic otitis media nowadays. The characteristic signs seem to have altered from the past ones. And the suspicion index is very low that there is often a considerable delay prior to diagnosis. This might result in irreversible complications such as hearing loss, facial nerve paralysis and labyrinthitis. It is difficult to be defined by culture, so many patients with tuberculous otitis media are subjected to have surgery without a correct etiologic diagnosis. Therefore bacteriologic study of secretion and pathologic examinations of biopsied tissue are mandatory. We report one year and 11 months old boy with tuberculous otitis media having painful otorrhea and perforated tympanic membrane diagnosed by histologic finding and polymerase chain reaction(PCR). And we would like to emphasize the fact that tuberculous otitis media must be considered in the differential diagnosis of persistent suppurative otitis media despite appropriate antibiotic therapy considering the increasing tendency in incidence of Tuberculosis in Korea these days.

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Effects of the Pharmacopuncture with MOK on Immune Regulation by Th1/Th2 Cytokines in L-Thyroxine-Induced Hyperthyroid Rats (L-thyroxine으로 유발된 갑상선기능항진증 랫트에서 MOK 약침의 Th1/Th2 사이토카인 조절에 의한 면역조절 효과)

  • Hwang, Ji Hye;Jung, Hyo Won
    • Korean Journal of Acupuncture
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    • v.35 no.2
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    • pp.91-97
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    • 2018
  • Objectives : This study was performed to investigate immune regulatory effects of the pharmacopuncture with MOK on hyperthyroid rats. Methods : The experimental hyperthyroidism was prepared by the intraperitoneal injection of L-thyroxine(LT4, 0.5 mg/kg) once daily for 2 weeks in Sprague-Dawley(SD) rats. The pharmacopuncture with MOK extract(MOK pharmacopuncture) at doses of 0.3 or 3 mg/kg was injected on acupuncture points in the thyroid glands of hyperthyroid rats once a day for 2 weeks. Propylthiouracil(PTU, 10 mg/kg) as a reference group was subcutaneously injected into the dorsal neck. We measured the levels of $IFN-{\gamma}$ and IL-4 in the sera of rats using enzyme-linked immunosorbant assay(ELISA) and determined the expression of $IFN-{\gamma}$, IL-4, IL-10, and Foxp3 in spleen tissues by reverse transcriptase-polymerase chain reaction(RT-PCR). Results : The treatment of MOK pharmacopuncture in hyperthyroid rats significantly decreased the serum levels of Th1 cytokine, $IFN-{\gamma}$(p<0.01 for MOK 0.3 mg/kg, p<0.05 for MOK 3 mg/kg, and p<0.05 for PTU) and significantly increased the levels of Th2 cytokine, IL-4(p<0.05 for MOK 0.3 mg/kg, p<0.001 for MOK 3 mg/kg, and p<0.05 for PTU) compared to control group. Also, the MOK pharmacopuncture significantly increased IL-4 expression(p<0.05 for MOK 3 mg/kg, and p<0.05 for PTU), IL-10(p<0.05 for MOK 3 mg/kg, and p<0.01 for PTU), and Foxp3(p<0.01 for MOK 0.3 mg/kg, p<0.05 for MOK 3 mg/kg and p<0.01 for PTU) in spleen tissues of hyperthyroid rats compared to control group. Conclusions : Our results suggest that MOK pharmacopuncture can help to ameliorate the pathological progression of hyperthyroidism by regulation of the Th1/Th2 imbalance.

Sequences and Phylogenic Analysis of Squid New Kinesin Superfamily Proteins (KIFs) (오징어과의 Kinesin Superfamily Proteins (KIFs)의 유전자분석 및 계통분석)

  • Kim, Sang-Jin;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.22 no.3
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    • pp.293-297
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    • 2012
  • The movement of vesicles from the neuronal cell body to specific destinations requires molecular motors. The squid giant axon represents a powerful model for studies of the axonal transport mechanism because the axoplasm can readily be separated from the sheath by simple extrusion. In a previous study, vesicular movements in the axoplasm of the squid giant axon were inhibited by the kinesin antibody. In the present study, we cloned and sequenced the cDNAs for squid brain KIFs. Amplification of the conserved nucleotide sequences of the motor domain by polymerase chain reaction (PCR) using first-strand cDNAs of the squid optic lobe identified six new KIF proteins. Motif analysis of the motor domains revealed that the squid KIFs are homologous to the consensus sequences of the mouse KIFs. The phylogenetic tree generated by using the maximum parsimony (MP) method, the neighbor-joining (NJ) method, the minimum evolution (ME) method, and the maximum likelihood (ML) method showed that squid KIFs are closest to mouse KIFs. These data prove the phylogenetic relationships between squid KIFs and mouse ones.

Use of Restriction Fragment Length Polymorphism Analysis to Differentiate Fungal Strains in Sunchang Meju

  • Jung, Jong-Hyun;Seo, Dong-Ho;Bhoo, Sung-Hee;Ha, Suk-Jin;Kim, Jong-Sang;Kim, Jeong-Hwan;Kwon, Dae-Young;Cha, Jae-Ho;Park, Cheon-Seok
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.888-891
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    • 2008
  • Twenty-three fungal strains were isolated from meju that had originated from the Sunchang province, the famous location for making fermented soybean foods in Korea. The restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rDNA (ITS-RFLP) was applied to differentiate the isolated fungal strains. First, the ITS region by polymerase chain reaction (PCR) with specific primers was amplified and then cleaved the products with different restriction enzymes. Cleavage of the amplified fragments with the restriction enzymes AluI, HaeIII, HhaI, and TaqI revealed extensive polymorphisms. The ITS-RFLP results highly correlated with ITS sequence analysis. All of the 23 fungal strains were classified into 5 groups by ITS-RFLP analysis. Aspergillus oryzae was the major fungal strain isolated from Sunchang meju (12 out of 23), while Aspergillus fumigatus was the next most frequently isolated strain (7 out of 23). In contrast, it was found that Fusarium asiaticum, Aspergillus sydowii, and Arthrinium sp. were the minor fungal strains in meju.

Differential Expression of Adipocyte-Related Molecules in the Distal Epididymal Fat of Mouse during Postnatal Period

  • Lee, Ki-Ho;Kim, Nan Hee
    • Development and Reproduction
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    • v.23 no.3
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    • pp.213-221
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    • 2019
  • The epididymal fat of mouse is a part of visceral fat deposit and is divided into the distal or proximal part based on its histochemical characteristics. Even though the formation of the epididymal fat pad begins before the birth, a detailed adipogenic procedure of the epididymal fat has not been revealed. The epididymal fat pad becomes enlarged and expended with age, and expressional changes of numerous genes are associated with the maturation of fat tissues. In the present research, expressional patterns of adipose tissue-related genes in the distal epididymal fat of mouse at 2, 5, 8, and 12 months of postnatal age were determined by a quantitative real-time polymerase chain reaction (PCR) analysis. The lowest transcript levels of fatty acid binding protein 4 (Fabp4), lipoprotein lipase (Lpl), delta like non-canonical Notch ligand 1 (Dlk1), peroxisome proliferator-activated receptor gamma (Pparg), leptin (Lep), adiponectin (Adipoq), and resistin (Retn) were detected at 2 months of age, except fatty acid synthase (Fasn) showing the lowest level at 5 months of age. Even though expression of Lep and Fabp4 were gradually increased until 12 months of age, significant increases of Pparg and Adipoq transcript levels were continued until 8 months of age. The transcript levels of Lpl, Rent, Dlk1, and Fasn were significantly increased at 8 months of age, compared with those at 2 months of age. The current findings suggest that the expansion of the distal epididymal fat of mouse during postnatal period would be companied with differential expression of various adipocyte-associated molecules.

The Effects of Gyejijakyakjimo-Tang on the Allergic Contact Dermatitis induced by DNCB (계지작약지모탕(桂枝芍藥知母湯)이 DNCB로 유발된 알레르기성 접촉피부염에 미치는 영향)

  • Kim, Sung-Ho;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.1
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    • pp.8-22
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    • 2010
  • Objectives : The purpose of this study is to investigate the effects of Gyejijakyakjimo-Tang on the Allergic Contact Dermatitis caused by 2,4-dinitro-chlorobezene(DNCB). Methods : Twenty eight mice were divided into four groups ; normal, control, experimental group A and B. Control and experimental groups were induced allergic contact dermatitis by DNCB. Experimental group A was orally administered the Gyejijakyakjimo-Tang and experimental group B was orally administered the prednisolone. In this study, ear thickness measurement, auricle microphotograph observation, MPO(Myeloperoxidase) activity measurement, Reverse transcription-polymerase chain reaction(RT-PCR) analysis of the mRNA level of TNF-$\alpha$, IL-$1{\beta}$ were performed on these four groups. In addition, the effect of Gyejijakyakjimo-Tang on cell viability and the effect of Gyejijakyakjimo-Tang on the compound 48/80-induced histamine release from HMC and RPMC were measured. Results: 1. Both experimental group A and B had decreased ear thickness compared with control group In contact hypersensitivity assay. 2. In experimental group A, inflammatory edema was similarly observed comparing to control group. Nevertheless, inflammatory edema was obviously reduced in experimental group B. In both experimental group A and B, pathological lesion of dermatitis were alleviated. In addition, the numbers of infiltrated inflammatory cells were decreased compared with control group. 3. Compared to the normal group, there was a noticeable increase in MPO activity in control group. However, in experimental group A and B, it showed remarkable inhibition of the increase in MPO activity comparing with control group. 4. The level of expression of TNF-$\alpha$, IL-$1{\beta}$ in experimental group A and B were meaningfully lower than those in control group. 5. In MTT assay, the concentrations of Gyejijakyakjimo-Tang that were used on the test had no cytotoxicity. 6. Gyejijakyakjimo-Tang dose-dependently inhibited the compound 48/80-induced histamine release from both HMC and RPMC. Conclusions : According to above experiments, Gyejijakyakjimo-Tang was effective on allergic contact dermatitis.

Immune-enhancing effects of polysaccharides with different molecular weights obtained from Gloiopeltis furcata (불등풀가사리 다당류의 분자량에 따른 면역증강 효과)

  • Lee, Dae-Hoon;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.25 no.1
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    • pp.1-6
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    • 2018
  • This study investigated the immune-enhancing effects of polysaccharides extracted from Gloiopeltis furcata (red seaweed) with different molecular weights. A crude polysaccharide mixture was hydrolyzed using acid treatment (0.1 N HCl) and three molecular weight fractions were generated and filtered using centrifugation: (${\leq}10kDa$, 10 to 100 kDa, and 100 kDa. Nitric oxide (NO) production in RAW264.7 cells treated with $0.01-0.5{\mu}g/mL$ polysaccharides ${\geq}100kDa$ was $12.28-19.05{\mu}M$. Treatment with polysaccharides ${\geq}100kDa$ increased cytokine levels, including TNF-${\alpha}$ and IL-6 levels, in a dose-dependent manner. Polymerase chain reaction analysis also revealed marked increases in iNOS and COX-2 mRNA expression levels. These findings lead us to conclude that macrophage activation induced by polysaccharides ${\geq}100kDa$ was greater than that induced by polysaccharides ${\leq}10kDa$ or between 10 and 100 kDa. The polysaccharides ${\geq}100kDa$ extracted from Gloiopeltis furcata investigated herein are potentially useful as natural immune-enhancing agents. These findings provided further insights into the potential use of ${\geq}100kDa$ as immunopotentiator or new function food.

Diagnosis and gI antibody dynamics of pseudorabies virus in an intensive pig farm in Hei Longjiang Province

  • Wang, Jintao;Han, Huansheng;Liu, Wanning;Li, Shinian;Guo, Donghua
    • Journal of Veterinary Science
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    • v.22 no.2
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    • pp.23.1-23.10
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    • 2021
  • Background: Pseudorabies (PR), caused by the pseudorabies virus (PRV), is an endemic disease in some regions of China. Although there are many reports on epidemiological investigations into pseudorabies, information on PRV gI antibody dynamics in one pig farm is sparse. Objectives: To diagnose PR and analyze the course of PR eradication in one pig farm. Methods: Ten brains and 1,513 serum samples from different groups of pigs in a pig farm were collected to detect PRV gE gene and PRV gI antibody presence using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Results: The July 2015 results indicated that almost all brain samples were PRV gE gene positive, but PRV gI antibody results in the serum samples of the same piglets were all negative. In the boar herd, from October 2015 to July 2018 three positive individuals were culled in October 2015, and the negative status of the remaining boars was maintained in the following tests. In the sow herd, the PRV gI antibody positive rate was always more than 70% from October 2015 to October 2017; however, it decreased to 27% in January 2018 but increased to 40% and 52% in April and July 2018, respectively. The PRV gI antibody positive rate in 100-day pigs markedly decreased in October 2016 and was maintained at less than 30% in the following tests. For 150-day pigs, the PRV gI antibody positive rate decreased notably to 10% in April 2017 and maintained a negative status from July 2017. The positive trend of PRV gI antibody with an increase in pig age remarkably decreased in three tests in 2018. Conclusions: The results indicate that serological testing is not sensitive in the early stage of a PRV infection and that gilt introduction is a risk factor for a PRV-negative pig farm. The data on PRV gI antibody dynamics can provide reference information for pig farms wanting to eradicate PR.

Molecular characterization of Malaysian fowl adenovirus (FAdV) serotype 8b species E and pathogenicity of the virus in specific-pathogen-free chicken

  • Sabarudin, Nur Syazana;Tan, Sheau Wei;Phang, Yuen Fun;Omar, Abdul Rahman
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.42.1-42.16
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    • 2021
  • Background: Inclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH. Objectives: To characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes. Methods: Suspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes. Results: The hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard. Conclusions: FAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.

Radiation Dose from Computed Tomography Scans for Korean Pediatric and Adult Patients

  • Won, Tristan;Lee, Ae-Kyoung;Choi, Hyung-do;Lee, Choonsik
    • Journal of Radiation Protection and Research
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    • v.46 no.3
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    • pp.98-105
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    • 2021
  • Background: In recent events of the coronavirus disease 2019 (COVID-19) pandemic, computed tomography (CT) scans are being globally used as a complement to the reverse-transcription polymerase chain reaction (RT-PCR) tests. It will be important to be aware of major organ dose levels, which are more relevant quantity to derive potential long-term adverse effect, for Korean pediatric and adult patients undergoing CT for COVID-19. Materials and Methods: We calculated organ dose conversion coefficients for Korean pediatric and adult CT patients directly from Korean pediatric and adult computational phantoms combined with Monte Carlo radiation transport techniques. We then estimated major organ doses delivered to the Korean child and adult patients undergoing CT for COVID-19 combining the dose conversion coefficients and the international survey data. We also compared our Korean dose conversion coefficients with those from Caucasian reference pediatric and adult phantoms. Results and Discussion: Based on the dose conversion coefficients we established in this study and the international survey data of COVID-19-related CT scans, we found that Korean 7-year-old child and adult males may receive about 4-32 mGy and 3-21 mGy of lung dose, respectively. We learned that the lung dose conversion coefficient for the Korean child phantom was up to 1.5-fold greater than that for the Korean adult phantom. We also found no substantial difference in dose conversion coefficients between Korean and Caucasian phantoms. Conclusion: We estimated radiation dose delivered to the Korean child and adult phantoms undergoing COVID-19-related CT examinations. The dose conversion coefficients derived for different CT scan types can be also used universally for other dosimetry studies concerning Korean CT scans. We also confirmed that the Caucasian-based CT organ dose calculation tools may be used for the Korean population with reasonable accuracy.