• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.157-165
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• 2001

Amplified fragment length polymorphism(AFLP) technique is based on the polymorphism detection through selective PCR amplification of restriction fragments from digested genomic DNA and thus includes the procedures of the total DNA digestion by endonucleases, ligation of adapters to the ends of the fragments, and following the selective amplification of the restricted DNA fragments. This study were aimed to : (1) determine the genetic variability of S aureus strains, (2) estimate genetic diversity within and among these strains, (3) compare phylogenetic relationships among these strains as genetic markers using AFLP techniques. Genomic DNA was digested with a particular combination of three restriction enzymes with specific recognition sites and the DNA fragments were ligated to restriction specific adapters and amplified using the selective primer combinations. In the S aureus strain, the number of scorable AFLP bands detected per each primer combination varied from 29 to 102, with an average of 61.59 using 27 primer combinations. A total of 1,663 markers were generated, 904 bands of which were polymorphic, showing a 33.48% level of polymorphism with these primer combinations. Among the primer combinations, E02/T02, E02/T03, E04/H02, E02/T01 and E04/H03 primer combinations showed a high level of polymorphism with 0.78, 0.76, 0.74, 0.71 and 0.70, respectively. But T03/H01, E01/T02 and E01/T03 primer combinations showed a low level of polymorphism with 0.38, 0.37 and 0.15, respectively, Therefore, the former primer combinations will be the most effective for AFLP analysis of S aureus. In SA1 sub-types the level of polymorphism of S aureus KCTC 1927 was similar to that of S aureus CU 01(0.825) and higher than those of other strains such as S aureus CU 02 (0.715), S aureus KCTC 2199(0.625), S aureus KCTC 1916(0.607) and S aureus KCTC 1621 (0.553). In SA2 sub-types the level of polymorphism of S aureus CU 07 was similar to that of S aureus CU 08(0.935) and higher than those of both S aureus CU 04(0.883) and S aureus CU 05(0.883) and lower than those of S aureus CU 03(0.583). In SA3 subtypes the level of polymorphism of S aureus CU 11 was similar to that of S aureus CU 12(0.913) and lower than that of S aureus CU 15(0.623). The results proved that AFLP marker analysis of S aureus strain could be used to study the epidemiology of mastitis and in addition, common genotype in geographic region could be useful for the development of an effective vaccine or DNA marker for easy diagnosis of mastitis caused by S aureus infection.

• Journal of Oral Medicine and Pain
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• v.34 no.4
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• pp.363-369
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• 2009

Erosive oral lichen planus (EOLP) and recurrent aphthous stomatitis (RAS) are T-cell mediated inflammatory immune disorders. It was investigated mRNA expression pattern of several regulatory factors, such as, CD28, CD45, CD152, CD154, CD279, which influence T lymphocyte in unstimulated whole saliva (UWS) of EOLP and RAS patients. It was collected unstimulated whole saliva during 10 minute in EOLP 18 people, RAS patients 12 people, healthy control 8 people. We investigated mRNA expression of T lymphocyte regulatory factors, such as, CD28, CD45, CD152, CD154, CD279, with real time reverse transcription polymerase chain reaction. In EOLP group, CD45, CD279 expressed higher and CD154 expressed lower than control. In RAS, CD45, CD270 expressed higher and CD28, CD154 expressed lower than control. In addition CD152 salivary mRNA expression of EOLP is higher than that of RAS. The above results were suggested that the mRNA expression of T lymphocyte regulatory factors in unstimulated whole saliva of EOLP and RAS contributes to diagnosis of diseases.

• Journal of Gastric Cancer
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• v.4 no.2
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• pp.109-120
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• 2004

Purpose: Individual gastric cancers demonstrate complicated genetic alterations. The PCR-based analysis of polymorphic microsatellite sequences on cancer-related chromosomes has been used to detect chromosomal loss and microsatellite instability. For the purpose of preoperative usage, we analyzed the correspondance rate of the microsatellite genotype between endoscopic biopsy and surgical specimens. Materials and Methods: Seventy-three pairs of biopsy and surgical specimens were examined for loss of heterozygosity and microsatellite instability by using 40 microsatellite markers on eight chromosomes. Microsatellite alterations in tumor DNAs were classified into a high-risk group (baselinelevel loss of heterozygosity: 1 chromosomal loss in diffuse type and high-level loss of heterozygosity: 4 or more chromosomal losses) and a low-risk group (microsatellite instability and low-level loss of heterozygosity: 2 or 3 chromosomal losses in diffuse type or $1\∼3$ chromosomal losses in intestinal type) based on the extent of chromosomal loss and microsatellite instability. Results: The chromosomal losses of the biopsy and the surgical specimens were found to be different in 21 of the 73 cases, 19 cases of which were categorized into a genotype group of similar extent. In 100 surgical specimens, the high-risk genotype group showed a high incidence of nodal involvement (19 of 23 cases: $\leq$5 cm; 23 of 24 cases: >5 cm) irrespective of tumor size while the incidence of nodal involvement for the low-risk genotype group depended on tumor size (5 of 26 cases: $\leq$5 cm; 18 of 27 cases: >5 cm). Extraserosal invasion was more frequent in large-sized tumor in both the high-risk genotype group ($\leq$5 cm: 12 of 23 cases; >5 cm: 23 of 24 cases) and the low-risk genotype group ($\leq$5 cm: 7 of 26 cases; >5 cm: 16 of 27 cases). The preoperative prediction of tumor invasion and nodal involvement based on tumor size and genotype corresponded closely to the pathologic tumor stage (ROC area >0.7). Conclusion: An endoscopic biopsy specimen of gastric cancer can be used to make a preoperative genetic diagnosis that accurately reflect the genotype of the corresponding surgical specimen.

• Tuberculosis and Respiratory Diseases
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• v.42 no.5
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• pp.654-659
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• 1995

Background: Tuberculosis has commonly been associated with various hematologic changes. A difference between the changes found in pulmonary tuberculosis and those found in miliary tuberculosis has been discussed. Up to now some worker were investigated hematological changes associated with pulmonary tuberculosis but was not investigated those associated with miliary tuberculosis in korea. Therefore we examimed the peripheral blood and bone marrow findings in miliary tuberculosis patients to determine hematologic changes. Methods: We performed blood sample at admission and bone marrow biopsy within 7days after admission. For evaluation of the hematologic findings, full blood counts and marrow differential counts were defined by the criteria outlined by Dacie and Lewis. Results: 1) Peripheral blood findings: Pancytopenia in 10% of patients, anemia in all patients, leukocytosis in 10% of patients, leukopenia in 20% of patients, thrombocytopenia in 30% of patients, lymphocytopenia in 90% of patients, monocytosis in 40% of patients and neutrophilia in 10% of patients were found at peripheral blood. 2) Bone marrow findings: Lymphocytopenia in 30% of patients, lymphocytosis in 20% of patients, plasmacytosis in 40% of patients, monocytosis in 100% of patients, and hypocellularity in 30% of patients were found at bone marrow. Erythropoiesis was decreased in 30% of patients. Granulopoiesis was decreased in 20% of patients and increased in 20% of patients. Bone morrow granuloma occured in 25% of patients. Conclusion: Hematologic changes of miliary tuberculosis were seen tendency of cytopenic pattern but monocyte was increased at peripheral blood and bone morrow. This findings would provide additional information for the differential diagnosis of miliary tuberculosis.

• Journal of Sasang Constitutional Medicine
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• v.10 no.2
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• pp.283-290
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• 1998

Sasang Constitutional Medicine focuses on the different constitutional manifestations of the individual's nature and emotions. The nature and emotions drive the ascending and descending of Qi in the body. And this dynamics of the Qi's ascent and descent shapes the different types of structures, functions and temperaments. Although Sasang Constitutional Medicine has many advantages, its diagnosis of the constitution still depends on the doctors' own idea and has no objective identification. So many doctors in Korea have been trying to solve this problem. Recently, there are several efforts to find out the relationship between genetic information and constitution. By the way, May, 1998 there is a astonishing report about the gene which determines the human performance, that is ACE(angiotensin converting enzyme). And it suggests that the I allele was associated with improved endurance performance. ACE has three genotype including II, ID and DD. "I" means insertion and "D" means deletion. We determined the type of the Sasang constitution with QSCCII questionaire and the one's ACE genotype with PCR of the 127 people and we discovered the relationship between the constitution and the ACE genotype. The result is as follow. Among 39 people who have the II genotype, 7(18%) belong to Taeum(Taiyin), 9(23.1%) belong to Soyang(Shaoyang) and 23(59%) belong to Soeum(Shaoyin). Among 62 people who have the ID genotype, 18(29%) belong to Taeum(Taiyin), 21(33.9%) belong to Soyang(Shaoyang) and 23(37.1%) belong to Soeum(Shaoyin). Among 26 people who have DD genotype, 11(42.3%) belong to Taeum(Taiyin), 4(15.4%) belong to Soyang(Shaoyang) and 11(42.3%) belong to Soeum(Shaoyin). This data indicates that there are implicable relationship between the Sasang constitution and ACE genotype. Especially people who have II genotype have much possibility to be a Soeum(Shaoyin) person (59%) and Soyang(Shaoyang) person have less possibility to have DD genotype (15.4%). With this conclusion, we suggest further study of relationship between the Sasang constitution and ACE genotype and we think that other polymorphism can be a candidate of the partner of Sasang constitution.

• Tuberculosis and Respiratory Diseases
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• v.69 no.6
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• pp.426-433
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• 2010

Background: Persistent cough has recently been found to be associated with Chlamydia pneumoniae infection. We aimed to investigate the infection rate of C. pneumonia in adult patients with chronic cough. Methods: We recruited 68 patients with persistent cough lasting in excess of 3 weeks, who visited Kangdong Sacred Heart Hospital from January 2005 to August 2005. On the first visit, chest and paranasal sinuses radiography, skin prick test of common allergens, and induced sputum samples for C. pneumoniae were performed in all of patients. Further evaluation for diagnosis included a methacholine provocation test and eosinophil counts in induced sputum. Results: The most common cause of chronic cough was upper airway cough syndrome (UACS) (26.5%), followed by eosinophilic bronchitis (20.6%) and cough variant asthma (16.2%). Idiopathic chronic cough was the cause in 33.8% of patients. The mean duration of cough was 11.7 months. C. pneumoniae was isolated by polymerase chain reaction (PCR) from one patient who had upper respiratory air way syndrome. Conclusion: Chlamydia pneumoniae appears to have a minor role as a cause of chronic cough in patients.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.363-370
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• 2014

Campylobacter jejuni is a prevalent foodborne pathogen worldwide. Human infection by C. jejuni primarily arises from contaminated poultry meats. Genes expressed in vivo may play an important role in the pathogenicity of C. jejuni. We applied an immunoscreening method, in vivo-induced antigen technology (IVIAT), to identify in vivo-induced genes during human infection by C. jejuni. An inducible expression library of genomic proteins was constructed from sequenced C. jejuni NCTC 11168 and was then screened using adsorbed, pooled human sera obtained from clinical patients. We successfully identified 24 unique genes expressed in vivo. These genes were implicated in metabolism, molecular biosynthesis, genetic information processing, transport, and other processes. We selected six genes with different functions to compare their expression levels in vivo and in vitro using real-time RT-PCR. The results showed that the selected six genes were significantly upregulated in vivo but not in vitro. In short, these identified in vivo-induced genes may contribute to human infection of C. jejuni, some of which may be meaningful vaccine candidate antigens or diagnosis serologic markers for campylobacteriosis. IVIAT may present a significant and efficient method for understanding the pathogenicity mechanism of Campylobacter and for finding targets for its prevention and control.

• Journal of Gastric Cancer
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• v.19 no.3
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• pp.301-314
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• 2019

Purpose: Peritoneal carcinomatosis in gastric cancer (GC) patients results in extremely poor prognosis. Malignant ascites samples are the most appropriate biological material to use to evaluate biomarkers for peritoneal carcinomatosis. This study identified exosomal MicroRNAs (miRNAs) differently expressed between benign liver cirrhosis-associated ascites (LC-ascites) and malignant gastric cancer-associated ascites (GC-ascites), and validated their role as diagnostic biomarkers for GC-ascites. Materials and Methods: Total RNA was extracted from exosomes isolated from 165 ascites samples (73 LC-ascites and 92 GC-ascites). Initially, microarrays were used to screen the expression levels of 2,006 miRNAs in the discovery cohort (n=22). Subsequently, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analyses were performed to validate the expression levels of selected exosomal miRNAs in the training (n=70) and validation (n=73) cohorts. Furthermore, carcinoembryonic antigen (CEA) levels were determined in ascites samples. Results: The miR-574-3p, miR-181b-5p, miR-4481, and miR-181d were significantly downregulated in the GC-ascites samples compared to the LC-ascites samples, and miR-181b-5p showed the best diagnostic performance for GC-ascites (area under the curve [AUC]=0.798 and 0.846 for the training and validation cohorts, respectively). The diagnostic performance of CEA for GC-ascites was improved by the combined analysis of miR-181b-5p and CEA (AUC=0.981 and 0.946 for the training and validation cohorts, respectively). Conclusions: We identified exosomal miRNAs capable of distinguishing between non-malignant and GC-ascites, showing that the combined use of miR-181b-5p and CEA could improve diagnosis.

• Korean Journal of Veterinary Service
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• v.42 no.1
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• pp.39-42
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• 2019

Tetanus is an acute, often fatal, and infectious disease of all species of domestic animals caused by the neurotoxin of Clostridium tetani (C. tetani). This disease is usually known to develop after microbial contamination in the deep or penetrating wound sites. In February 2017, a farmer who was raising 76 cows injected foot and mouth disease vaccine to three or more cows with one syringe. Their clinical symptoms were observed 2 to 16 days after the vaccination. The initial symptoms were stiffness, rigidity of the neck and limbs, pricked ears, and prolapse of the third eyelid. Subsequently, there was recumbency with extension of the limbs, convulsions and opistotonus and the affected 20 cows were all died. Two dead cows were submitted to Animal and Plant Quarantine Agency for disease diagnosis. At necropsy, a focal edematous abscess of 15 to 20 cm in diameter was grossly observed in the subcutaneous and intramuscular tissue of scapular region and filled with a large amount of greenish pus. The feed was full in oral cavity and slightly observed in the trachea and lobes of lung. Histopathologically, focal granulomatous nodules with eosinophilic materials in the tissue were observed. In the lung, aspiration pneumonia and gram (+) bacteria were seen. The C. tetani was isolated in samples anaerobically cultured using reinforced clostridial medium and identified by PCR. To our knowledge, no previous outbreak of tetanus in cattle has affected such a high number of animals; neither has it been associated with misuse of the same syringe and needle to administer multiple individuals.

• Korean Journal of Clinical Laboratory Science
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• v.53 no.1
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• pp.41-48
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• 2021

Essential thrombocythemia (ET) is a clonal bone marrow stem cell disorder, primarily involving the megakaryocytic lineage. The WHO 2016 guidelines include the molecular detection of JAK2, MPL, and CALR mutations as a major diagnostic criterion for ET. This study aimed to determine the frequency of JAK2 V617F, MPL W515L, and CALR mutations in Iraqi Kurdish patients afflicted with ET, and to analyze their clinical and hematological features. A total of 73 Iraqi Kurdish patients with ET were enrolled as subjects, and analysis was achieved utilizing real-time PCR. The frequency of JAK2 V617F, CALR, and MPL W515L mutations was determined to be 50.7%, 22%, and 16.4%, respectively. No statistically significant difference was obtained when considering the age and gender among different genotypes. The JAK2 V617F mutated patients had significantly higher white blood cell counts and hemoglobin levels than the CALR-positive patients (P-value=0.000, 0.007, respectively), MPL W515L-positive patients (P-value=0.000, 0.000, respectively), and triple negative patients (P-value=0.000, 0.000, respectively). Also, the JAK2 V617F mutated patients showed higher platelet count as compared to the MPL W515L-positive patients (P-value=0.02) and triple negative patients (P-value=0.04). Furthermore, significantly lower white blood cell count and hemoglobin levels were associated with CALR positivity (P-value=0.000, 0.01, respectively), MPL W515L-positivity (P-value=0.001, 0.000, respectively), and triple negativity (P-value=0.000, 0.000, respectively), as compared to patients with combined mutations. In conclusion, apart from a relatively high frequency of MPL W515L mutation, our data is comparable to earlier reports, and highlights the importance of genotyping the JAK2 V617F, MPL W515L, and CALR mutations for accurate diagnosis of patients with ET.