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Development of a Duplex RT-PCR Assay for the Simultaneous Detection and Discrimination of Avirulent and Virulent Newcastle Disease Virus (NDV) (뉴캣슬병 바이러스 검출 및 병원성 감별을 위한 Duplex RT-PCR법 개발)

  • Kim, Ji-Ye;Lee, Hyun-Jeong;Jang, Il;Lee, Hee-Soo;Yoon, Seung-Jun;Park, Ji-Sung;Seol, Jae-Goo;Kim, Seung-Han;Hong, Ji-Mu;Wang, Zillian;Liu, Hualei;Choi, Kang-Seuk
    • Korean Journal of Poultry Science
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    • v.44 no.2
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    • pp.93-102
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    • 2017
  • A duplex RT-PCR (dRT-PCR) assay was developed for the simultaneous detection and discrimination of non-virulent and virulent Newcastle disease virus (NDV) in a single PCR tube. Primers targeting the large polymerase protein (L) gene and the fusion protein (F) gene of NDV were designed to detect all NDVs (by common type PCR primers) and virulent NDVs (by pathotype PCR primers), respectively and evaluated experimentally with reference NDV strains and other poultry viral pathogens. PCR products of the expected size of 386 bp were amplified from all NDV samples whereas PCR products of the expected size of 229 bp were amplified from virulent NDV samples alone. Cross reaction was not observed with other avian viral pathogens. The detection limit of NDV by the dRT-PCR was estimated to be $10^3$ 50% egg infectious dose/0.1 mL. In the dRT-PCR using field isolates of NDV, the pathotype PCR primers detected specifically all of virulent field isolates of NDV from Malaysia, Pakistan and China whereas common type PCR primers detected 94.4% (51/54) of field isolates of NDV from China. Three Chinese NDV isolates with false negative result were non-virulent viruses. Our results indicate that the dRT-PCR might provide a rapid and simple tool for rapid simultaneous detection and discrimination of non-virulent and virulent NDVs. Therefore the developed dRT-PCR assay provides a powerful novel means for the rapid diagnosis of Newcastle disease.

Gastric Dilatation and Volvulus in Shih-tzu (시츄에서 발견된 위확대염전)

  • Moon, Joon Ho;Kim, Hyun Ah;Ryu, Minok;Jang, Min;Ji, Seoyeon;Lee, Inhyung;Yoon, Junghee;Choi, Mincheol;Youn, Hwayoung;Lee, Byeong Chun;Jang, Goo
    • Journal of Veterinary Clinics
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    • v.32 no.3
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    • pp.255-258
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    • 2015
  • Gastric dilatation and volvulus (GDV) is an acute and life-threatening disease most commonly affecting large- and giant-breed dogs. However a 17-year-old Shih-tzu (4 kg, spayed female) was hospitalized for acute GDV. Repeated unproductive retching, lethargy, and excessively enlarged abdomen were observed. Physical examination indicated that the patient had suffered from hypothermia ($36.5^{\circ}C$), tachycardia (240 bpm), slowed capillary refill time (> 2 sec.), and pale mucous membrane. Grade III murmur with normal lung sound was auscultated. Abdominal palpation revealed that tympanic regions existed in both the left and right sides. Systolic blood pressure decreased gradually from 220 to 40 mmHg within 4 hours. In blood analysis, slight azotemia was observed by blood urea nitrogen (BUN; 29.1 mg/dl) and creatinine (1.6 mg/dl). Blood lactate concentration (8.13 mmol/l) was severely elevated. Additionally, dilatation and volvulus of the stomach was observed by radiograph. Supportive oxygen, heat, fluid, and drugs were administered with gastric decompressions (e.g., gastrocentesis and nasogastric tube). However the patient entered into comatose status with uncontrollable systolic blood pressure, despite the administration of dobutamine intravenously. The case was closed by euthanasia, considering welfare and age. We finally diagnosed the patient as a GDV, thus this is the first GDV case report in small-breed dog such as Shih-tzu.

Differences in Clinical Manifestations and Treatment Responses in Influenza Type A and B in a Single Hospital during 2013 to 2015 (단일 병원에서 2013-2015년에 유행한 인플루엔자 A형과 B형의 임상 양상 및 치료반응의 차이)

  • Lee, Sang Min;Park, Sang Kyu;Kim, Ji Hyun;Lee, Jung Ha;Na, So Young;Kim, Do Hyun;Kang, Eun Kyeong;Cho, Sung Min;Kim, Hee Sub
    • Pediatric Infection and Vaccine
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    • v.24 no.1
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    • pp.16-22
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    • 2017
  • Purpose: We suspect there is a difference in the clinical manifestations and treatment response to antiviral drugs for influenza A and B. This study was conducted to investigate this difference. Methods: We collected information on pediatric patients, infected with the influenza virus, admitted to Dongguk University Ilsan Hospital from October 2013 to May 2015. We investigated the clinical manifestations of influenza and differences in treatment response to oseltamivir treatment for the two types of influenza. Results: A total of 138 patients were included. The mean age was $3.5{\pm}4.0$ years. When comparing the diseases associated with influenza A and B, croup (19.2% vs. 1.7%, P=0.001) was more common with influenza A infection. Myositis (0% vs. 6.7%, P=0.021) and gastroenteritis (29.5% vs. 46.7%, P=0.038) were more common with influenza B infection. When comparing the total fever duration from the start of oseltamivir administration, patients treated with oseltamivir within 2 days of fever had the shortest duration. Among the patients treated with oseltamivir, the duration of fever, after the start of oseltamivir treatment, for was shorter for influenza A infection than for influenza B infection ($16.0{\pm}19.1$ hours vs. $28.9{\pm}27.9$ hours, P=0.006). Conclusions: There appear to be differences in the accompanying diseases and antiviral medication responses between the two types of influenza. It is important to administer oseltamivir within 2 days of fever.

Investigation of radical scavenging and anti-inflammatory activity of the extract and fractions of Termitomyces albuminosus (계종버섯 추출물과 분획물의 라디칼 소거 활성과 항염증 활성 분석)

  • Kang, Min-Jae;Kim, Young-Jae;Hwang, Hyeon-Jeong;Kim, Moon-Ho;Sin, Jong-Moo;Shin, Pyung-Gyun;Kim, Gun-Do;Kim, Keun Ki
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.45-52
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    • 2020
  • Termitomyces albuminosus has been recognized to have the best mushrooms in China, in terms of taste and aroma. The efficacy of these mushrooms has been recorded in the botanical list. However, research on the development of their artificial culture methods is necessary. In this study, we prepared an organic solvent extract and a hot water extract to understand the development of compounds and functional foods with antioxidant and anti-inflammatory activities. The IC50 value of DPPH radical scavenging activity of the hot water extract (TA4) was 1.5 mg/mL and the IC50 value of the MeOH fraction (TA2) was 1.93 mg/mL. The anti-inflammatory activity was investigated by the inhibition of NO production. EtOAc fraction (TA1) is a crude extract, but 79% of NO production was inhibited at 100 ㎍/mL. NO was not produced at 200 ㎍/mL. TA1-5-6, from TA1 inhibited NO production by 15% as compared to the positive control at 15 ㎍/mL, and completely inhibited NO production at 30 ㎍/mL. No cytotoxicity was observed at 50 ㎍/mL. TA2-1-5 from the MeOH fraction (TA2) inhibited more than 75% of NO production at 30 ㎍/mL; cytotoxicity was very low even at 50 ㎍/mL. In conclusion, by selective solvent selection, it was possible to manufacture an extract with no cytotoxicity and excellent biological activities. Furthermore, the extracts showed potential for developing various functional foods and drugs.

Effect of Acute and Chronic Treatment with Risperidone on the Serotonin and Dopamine Receptors in the Rat Brain (Risperidone의 급성 및 만성 투여가 흰쥐 뇌의 Serotonin과 Dopamine 수용체에 미치는 영향)

  • Choi, Yun-Young;Son, Hye-Kyung;Kim, Chang-Yoon;Lee, Chul;Lee, Hee-Kyung;Moon, Dae-Hyuk
    • The Korean Journal of Nuclear Medicine
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    • v.31 no.1
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    • pp.9-18
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    • 1997
  • The therapeutic efficacy of antipsychotic drugs is generally attributed to their ability to block dopamine $D_2$ receptors. Classical $D_2$ antagonists are not effective to treat negative symptoms and produce extrapyramidal side effects On the other hand, atypical antipsychotic agents ameliorate negative symptoms without producing extra-pyramidal side effects, and it is reported to be associated with blockade of serotonin $5-HT_2$ receptors. The purpose of this study was to evaluate the effect of risperidone on neuroreceptors in the rat brain by Quantitative autoradiography method. In acute treatment group, risperidone was injected into Peritoneal cavity of male Wistar rats with dose of 0, 0.1, 0.25, 0.5, 1.0 and 2.0mg/kg in each group(5/group), and they were decapitated after 2 hours. In chronic treatment group, risperidone was injected with dose of 0, 0.1, and 1mg/kg(I.P.) for 21 days and decapitated after 24 hours following last treatment. The effect of risperodone on the binding of [$^3H$]spiperone to $5-HT_2$ and $D_2$ receptors were analysed in 4 discrete regions of the striatum, nucleus accumbens, and frontal cortex by quantitative autoradiography Acute treatment with risperidone reduced cortical $5-HT_2$ specific [$^3H$]spiperone binding to 32% of vehicle-treated control. Subcortical $5-HT_2$ specific [$^3H$]spiperone binding was not affected at all dose groups whereas a significant reduction (57%) in $D_2$ specific [$^3H$]spiperone binding was observed in risperidone treated group at doses of 1-2mg/kg. Chronic treatment with risperidone produced a decrease in the maximal number of cortical $5-HT_2$ receptors to 51% and 46% of control in 0.1mg/kg & 1mg/kg treated group respectively. In conclusion, risperidone is a cortical serotonin receptor antagonist with relatively weak antagonistic action on dopamine receptors. These effects oil neuroreceptors may explain the therapeutic effect of risperidone as a atypical antipsychotic agents.

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Monitoring Methylmercury in Abyssal Fish (심해성 어류 중 메틸수은 모니터링)

  • Kim, Seong-Cheol;Jang, Jin-Wook;Kim, Hyun-Ah;Lee, Sang-Ho;Jung, Young-Ji;Kim, Ji-Yeon;Ahn, Jong-Hoon;Park, Eun-Hye;Ko, Yong-Seok;Kim, Dong-Sul;Kim, Sang-Yub;Jang, Young-Mi;Kang, Chan-Soon
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.383-389
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    • 2010
  • The aim of this study was to determine the methylmercury (MeHg) levels in abyssal fish species. The MeHg in the fishes was extracted with hydrochloric acid and toluene and then purified using an L-cysteine solution. The extract was analyzed with a gas chromatography-electron capture detector (GC-${\mu}ECD$) with a thermon Hg-capillary column. The detection limit and the recovery of the method were 0.002 and 84.2-98.5% (mean, 93.4%), respectively. The MeHg content in 492 abyssal fishes ranged from 0.037 to 2.009 mg/kg. The levels of MeHg [range, mg/kg (mean)] were significantly dependent on fish species and presented as the following; 0.157-2.009 (0.546) in Scalloped hammerhead shark, 0.211-0.878 (0.501) in Blue shark, 0.121-0.993 (0.482) in Spiny dogfish, 0.243-0.658 (0.397) in Salmon shark, 0.074-1.958 (0.353) in Blacktip shark, 0.038-0.807 (0.302) in Southern hake, 0.099-0.511 (0.300) in Scorpion fish, and 0.037-0.133 (0.067) in Ling. The monitoring results showed that the estimated weekly intake of MeHg from sharks, Southern hake, and Ling were lower than the provisional tolerable weekly intake recommended by the Joint FAO/WHO expert committee on food additives.

Induction of c-Jun Expression by Breast Cancer Anti-estrogen Resistance-3 (BCAR3) in Human Breast MCF-12A Cells (정상적인 인간유방상피세포인 MCF-12세포에서 유방암 항에스토젠 내성인자-3 (BCAR3)에 의한 c-Jun 발현 유도 연구)

  • Oh, Myung-Ju;Kim, Ji-Hyun;Jhun, Byung Hak
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1383-1391
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    • 2016
  • Anti-estrogen drugs such as tamoxifen have been used for treating patients with ER-positive, early breast cancer. However, resistance to anti-estrogen treatment is inevitable in most patients. Breast cancer anti-estrogen resistance-3 (BCAR3) has been identified as the protein responsible for the induction of tamoxifen resistance in estrogen-dependent human breast cancer. We have previously reported that BCAR3 regulates the cell cycle progression and the signaling pathway of EGF and insulin leading to DNA synthesis. In this study, we investigated the functional role of BCAR3 in regulating c-Jun transcription in non-tumorigenic human breast epithelial MCF-12A cells. A transient transfection of BCAR3 increased both the mRNA and protein of c-Jun expression, and stable expression of BCAR3 increased c-Jun protein expression. The overexpression of BCAR3 directly activated the promoter of c-jun, AP-1, and SRE but not that of $NF-{\kappa}B$. Furthermore, single-cell microinjection of BCAR3 expression plasmid in the cell cycle-arrested MCF-12A cells induced c-Jun protein expression, and co-injection of dominant negative mutants of Ras, Rac, and Rho suppressed the transcriptional activity of c-Jun in the presence of BCAR3. Furthermore, stable expression of BCAR3 increased the proliferation of MCF-12A cells. The microinjection of inhibitory materials such as anti-BCAR3 antibody and siRNA BCAR3 inhibited EGF-induced c-Jun expression but did not affect IGF-1 induced upregulation of c-Jun. Taken together, we propose that BCAR3 plays a crucial role in c-Jun protein expression and cell proliferation and that small GTPases (e.g., Ras, Rac, and Rho) are required for the BCAR3-mediated activation of c-Jun expression.

Antiglycemic Effect of Carnosine in Diabetic Mice (당뇨 마우스에서 카르노신의 혈당강하 효과)

  • Hue, Jin-Joo;Kim, Jong-Soo;Kim, Jun-Hyeong;Nam, Sang-Yoon;Yun, Young-Won;Jeong, Jae-Hwang;Lee, Beom-Jun
    • Journal of Food Hygiene and Safety
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    • v.24 no.4
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    • pp.391-397
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    • 2009
  • Carnosine is a dipeptide ($\beta$-alanyl-L-histidine) found in mammalian brain, eye, olfactory bulb and skeletal muscle at high concentrations. Its biological functions include antioxidant and anti-glycation activities. The objectives of this study were to investigate anti-diabetic effects of carnosine as determined by blood glucose levels in glucose tolerance test (GTT) and insulin tolerance test (ITT), insulin level and serum biochemical and lipid levels in male C57BL/6J db/db mice. There were five experimental groups including normal (C57BL/6J), control (vehicle), and three groups of carnosine at doses of 6, 30, and 150 mg/kg b.w. Carnosine was orally administered to the diabetic mice everyday for 8 weeks. There was no significant difference in body weight changes in carnosine-treated groups compared to the control. The treatments of carnosine significantly decreased the blood glucose level in the diabetic mice compared with the control (p < 0.05) after 5 weeks. The treatments of carnosine also significantly decreased the blood glucose levels in GTT and ITT and glycosylated hemoglobin (HbA1c), compared with the control (p < 0.05). Carnosine at the dose of 6 mg/kg significantly decreased the serum insulin level compared to the control (p < 0.05). Carnosine significantly increased total proteins but significantly decreased lactate dehydrogenase and blood urea nitrogen compared with the control (p < 0.05). Carnosine also significantly decreased glucose, LDL, and triglyceride in the serum of diabetic mice compared to the control (p < 0.05). These results suggest that carnosine has a hypoglycermic effect resulting from reduction of glucose and lipid levels and that high carnosine-containing diets or drugs may give a benefit for controlling diabetes mellitus in humans.

Therapeutic Efficacy of Lamivudine in Children and Adolescents with Chronic Hepatitis B (만성 B형 간염 소아청소년 환자에서의 라미부딘 치료 효과)

  • Choi, Yujung;Bae, Kil Seoung;Kim, Ki Hwan;Koh, Dae Kyun;Kim, Jong-Hyun
    • Pediatric Infection and Vaccine
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    • v.25 no.2
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    • pp.72-81
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    • 2018
  • Purpose: This prospective study aimed to investigate the therapeutic efficacy of lamivudine in children with chronic hepatitis B virus (HBV) infection. Methods: During July 2003 through October 2015, children with chronic hepatitis B who visited our institution were included in this study. Fifty-five patients, who received first-line treatment of lamivudine (3 mg/kg, 100 mg maximum) for over three months, were enrolled. After initiating lamivudine, alanine aminotransferase (ALT), HBV-DNA, and HBV markers were followed up at 1 month, 3 months, and every 3 months, thereafter. The treatment endpoint was determined as 1) normalization of ALT, 2) HBeAg seroconversion, and 3) anti-HBe positivity for twelve consecutive months. Results: Thirty-one male (56.4%) and 24 female (43.6%) patients were included. The mean age at treatment initiation was 8.1 years. The mean duration of treatment was 23.4 months. ALT normalization was found in 98.2% (54 of 55). Anti-HBe seroconversion was found in 70.6% (36/51). Loss of HBsAg was found in 10.9% (6/55). All biochemical responses occurred under age seven. The rate of virologic response (defined as HBV-DNA <2,000 IU/mL) at six months after treatment initiation was 78.7% (37/47). At twelve months after reaching treatment endpoint, 87.2% (34/39) maintained their virologic response. Resistance to lamivudine was found in 16.4% (9/55). Conclusions: Lamivudine treatment in Korean pediatric patients with chronic hepatitis B showed better outcomes compared with other studies that implemented similar protocols in foreign populations. Further studies are needed to investigate the efficacy of newly recommended antiviral drugs on the Korean pediatric population.

A Literal Study of Feature of the Preventive Medicine in Oriental Medical Science (한의(韓醫) 예방의학(豫防醫學)의 특징(特徵)에 관(關)한 문헌적(文獻的) 고찰(考察))

  • Lee Sang-Woon;Lee Sun-Dong
    • Journal of Society of Preventive Korean Medicine
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    • v.1 no.1
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    • pp.85-104
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    • 1997
  • As the concern about health is increased, the importance of preventive medicine that prevent disease in advance to overcome boundary of disease remedy gets emphasized in the whole world. The fundamental thoughts of oriental medicine are the harmony between the human body and nature, the unified idea regarding mind in the same light with body, and the symmetry of the cosmo dual forces. And oriental medicine is a kind of study that has developed on the ground of prevention thought. from old days, it has been developed the preventive medicine at is called 'Yangseng(養生; recuperation)' etc. with the clinical medicine. The preventive medicine of oriental medicine was taking a serious of the first preventive medicine of an incurable disease thought and the prevention thought that belongs to the second and third preventive medicine is described through the whole oriental medicine. Also the clinical and preventive medicine to apply to the clinics have an in- separable relation. Therefore I inspected the part of the preventive medicine described in some oriental medicine books and studied the characteristics of preventive medicine of oriental medicine as follows; First, the preventive medicine of oriental medicine has the characteristics that is emphasized generally in the first preventive medicine and wholly it is composed in the system of the first, second and third preventive medicine. It has the presentative theory such as 'Jungkijonae sabulkaghan(正氣存內 邪不可干; If good energy is in body, a disease. dosen't occur)', 'Husajukpung Phijeyushi(虛邪賊風 避之有時; When e infectious disease like plague break out, they must avoid the place occurring the disease)', 'Chuninsangeung(天人相應; The harmony of nature and human)' etc.. This is intimately related to the incurable disease thought of the inside diameter and oriental medicine has pursued that. Second, due to the most prerequisite theory of disease production, the balanced condition of environment, the cause of a disease and host is called the health. As oriental medicine has the system of aetiology like that, we can see the host and environments are importance most of all. Namely we can think of the relation of host >> environment > the cause a disease Up to date Jungkijonae Sabulkaghan(正氣存內 邪不可干) that the most oriental medicine doctors have had a knowledge is not whole theory but a pan of oriental medicine science and it is included in oriental medicine theory to avoid infectious disease such as Husajukpung Phijeyushi(虛邪賊風 避之有時). Third, according to the natural result of the first and second contents, we can know that its characteristics stress the remedy without drugs. Because Jungkijonae Sabulkaghan(正氣存內 邪不可干), Husasukpung Phijeyushi(虛邪賊風 避之有時), and Chuninsangeung(天人相應) mean that they prevent disease in condition of freedom from ailment, We can prevent the disease production through the positive preventive methods such as exercise, spirit, food and innate prevention etc.. fourth, the preventive medicine of oriental medicine has developed with therapeutics and it contains all oriental medical methods such as host, environment, exercise, acupuncture and moxibustion, innate or postnatal methods. Also it is the general preventive medicine that has fundamental philosophy of oriental medicine; for examples, the unified idea, the harmony of nature and human, and the unity of mind and body. fifth, to develope the above scientific merit and special features, the preventive medicine of Oriental medicine must be objectified and reemergent stud? gets more and more essential from now on. Especially we need to have the scientific concern of Oriental medicine about the cause of a disease, environmental hygiene, industrial sanitation, and personal hygiene etc..

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