• Title/Summary/Keyword: 바이러스 증식

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bovine leukosis (산재성 송아지 백혈병)

  • 임금기;장현철;강문일
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2002.11a
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    • pp.150-150
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    • 2002
  • 40두 규모의 농장에서 사육된 홀스타인 종, 4개월령의 수컷 송아지에서 거세 후 고열, 식욕부진, 기침 등의 증상과 함께 전신 체표 림프절의 종대가 관찰되어 백혈병으로 잠정진단 하고 도태를 권유하였으나 축주가 치료를 원해 항생제와 해열제 및 기타 대증요법을 실시한 후 치료반응이 없어 폐사하여 부검을 실시하였다. 혈액 검사상 이상핵과 다형핵을 가진 다수의 림프구 및 백혈구, 호중구, 림프구 및 단핵구의 증가가 관찰되었다. 또한 백혈병 바이러스에 대한 분리 및 PCR 검사는 음성이었다. 부검 소견으로 체표 림프절, 슬관절 부위 및 비장과 간의 종대가 관찰되었으며 비장 중심에 12x11 cm의 종괴와 폐의 전엽부 유착 및 폐문, 종격동 림프절의 심한 종대가 관찰되었다. 견갑전 림프절을 비롯하여 대퇴골 전, 서혜, 폐문, 이하림프절 등 전신 림프절의 종대 및 소성의 연한 황색의 매끄러운 절단면을 보였다. 전지 관절의 종대와 관절강 내부는 증가된 농성 활액을 보였으며 고관절 강 내에 농성 활액의 증가와 공기 노출 후 젤리양 응고를 보였다. 심장은 장액성 위축과 함께 섬외막성 점상출혈이 나타났다. 병리조직학적 소견으로 비장은 지주 주변에 미성숙형의 세포들의 침윤이 보이며 유사분열상이 다수 관찰되었고 백색 수질에도 유사분열상의 증가와 함께 림프아구성 세포들이 다수 나타났다. 이들 주요한 비정상 세포들은 다형성의 큰 핵을 가진 다양한 림프아구의 형태를 지녔으며 핵내 공포가 인정되었다. 비장의 종괴 주변에는 증식된 섬유조직으로 둘러싸여 있었으며 미세농양 형성되어 있고, 일부 석회화가 진행된 부위도 있었다. 간소엽성 중심성 울혈과 가벼운 간세포내 지방침윤, 혈관 내 림프아구 형태의 세포와 소수의 호중구가 관찰되었다. 간삼조 주변에는 가볍거나 중등도의 단핵세포의 침윤이 미만성으로 관찰되었다. 폐에서는 중등도의 기관지 폐렴과 함께 일부는 무기폐가 관찰되었으며 폐포강과 세기관지내에는 염증성 삼출물이 다량 들어 있었다. 다병소성 미세농양과 함께 괴사가 있었고 실질의 섬유화가 진행되어 있었다. 또한 중등도의 간질성 신장염과 림프절은 지주 주변에 간극 내 비정상 림프구 세포의 형태는 비장의 그것과 유사하였으며 적수와 백수의 구이 힘들며 림프소절이 증가되어 있었다. 한 시야에서 유사분열상이 6-8 개로 그 지수가 매우 높으며 이와 더불어 큰 림프구가 전반에 걸쳐 침윤되어 있었다. 주변부 동(sinus)에는 많은 물질들이 침윤되어 있으며 렴프소절내 미만성의 성상현상이 관찰되었다. 회장은 파이어판내 심한 림프구 소실이 나타났다. 이상의 소견을 바탕으로 본 증례는 산재성 송아지 백혈병으로 진단되었다

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Effects of Several Herbal Medicines on the Replication of Hepatitis B Virus (수종(數種)의 한약재(韓藥材)가 B형 간염(肝炎)바이러스 증식억제(增殖抑制)에 미치는 효과(效果))

  • Cho, Hong-Kun;Ahn, Duk-Kyun;Lee, Song-Deuk
    • The Journal of Korean Medicine
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    • v.19 no.2
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    • pp.244-270
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    • 1998
  • This study was performed to investigate an anti-HBV activities of the aqueous extracts from 10 Korean herbal medicines in the HepG2 2.2.15 cell culture system and the results were as follows: 1. The extracts of 6 plants (Herba Artemisiae Capillaris, Radix et Rhizoma Rhei, Cortex Cinnamomi, Fructus Chebulae, Fructus Rubi and Radix Rubi) decreased, significantly and dose-dependently, the levels of extracellular HBV virion in the concentrations (10, 100, 500 and $1,000\;{\mu}g/m{\ell}$) tested. 2. However, others (Radix lsatidis, Lignum Sappan, Herba Lysimachiae and Fructus Lycii) did not show any effect either on the replication of HBV or on the levels of virion DNA in the culture media of HepG2 2.2.15 cell. 3. Among the 6 plants which showed the inhibitory potency on the production of extracellular HBV virion, Radix et Rhizoma Rhei, Cortex Cinnamomi, Fructus Chebulae, Fructus Rubi and Radix Rubi except Herba Artemisiae Capillaris also showed the inhibition of the replication of intracellular HEV DNA in the range of $100{\sim}500\;{\mu}g/m{\ell}$. Considering the above results, it is thought that 6 plants(Herba Artemisiae Capillaris, Radix et Rhizoma Rhei, Cortex Cinnamomi, Fructus Chebulae, Fructus Rubi and Radix Rubi) possess the anti-HBV activities in the HepG2 2.2.15 cell culture system. We thus suggest that these plants possess a potential as a therapeutic agent for the chronic viral hepatitis. These results might be useful as a basic data for the development of the new preventive drugs for HBV diseases.

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Extract of Linum usitatissimum L. inhibits Coxsackievirus B3 Replication through AKT Signal Modulation (아마인 추출물의 AKT 신호 조절을 통한 콕사키바이러스 증식억제)

  • Shin, Ha-Hyeon;Moon, Sung-Jin;Lim, Byung-Kwan;Kim, Jin Hee
    • Korean Journal of Pharmacognosy
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    • v.49 no.4
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    • pp.291-297
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    • 2018
  • Coxsackievirus B3 (CVB3) is a very well-known causative agent for viral myocarditis and meningitis in human. However, the effective vaccine and therapeutic drug are not developed yet. CVB3 infection activates host cell AKT signaling. Inhibition of AKT signaling pathway may attenuate CVB3 replication and prevent CVB3-mediate viral myocarditis. In this study, we determined antiviral effect of the selected natural plant extract to develop a therapeutic drug for CVB3 treatment. We screened several chemically extracted natural compounds by using HeLa cell-based cell survival assay. Among them, Linum usitatissimum L. extract was selected for antiviral drug candidate. L. usitatissimum extract significantly decreased CVB3 replication and cell death in CVB3 infected HeLa cells with no cytotoxicity. CVB3 protease 2A induced eIF4G1 cleavage and viral capsid protein VP1 production were dramatically decreased by L. usitatissimum extract treatment. In addition, virus positive and negative strand genome amplification were significantly decreased by 1 mg/ml L. usitatissimum extract treatment. Especially, L. usitatissimum extract was associated with inhibition of AKT signal and maintain mTOR activity. In contrast, Atg12 and LC3 expression were not changed by L. usitatissimum extract treatment. In this study, the potential AKT signal inhibitor, L. usitatissimum extract, was significantly inhibited viral genome replication and protein production by inhibition of AKT signal. These results suggested that L. usitatissimum extract is a novel therapeutic agent for treatment of CVB3-mediated diseases.

trans-Cinnamaldehyde-Induced Apoptosis in AGS Cells (AGS 세포주에서 트랜스 신남알데하이드의 세포 사멸 유도)

  • Lee, Sunyi;Jung, Joohee
    • Journal of Food Hygiene and Safety
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    • v.36 no.1
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    • pp.100-104
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    • 2021
  • trans-Cinnamaldehyde (TCA), as one of the active ingredients in cinnamon, has been reported to have antiviral, antibacterial and antifungal effects as well as anti-cancer effects in several cancer cell lines. However, reports of TCA in gastric cancer are rare, and its mechanism is unclear. In this study, we investigated the anti-proliferative effect of TCA and its mechanism in gastric cancer AGS cells. TCA dose-dependently inhibited the cell viability of AGS cells. Our results suggested that TCA induces apoptosis through changes in cell morphology. To elucidate its mechanism, we investigated the expression level of apoptosis-related proteins. TCA induced the expression of p53 and Bax proteins, and then increased the cleaved caspase 9 and cleaved PARP. These results indicated that TCA triggers apoptosis via p53 pathway in AGS cells. Our results suggested that TCA might be a new anticancer drug candidate for gastric cancer.

Thrips Infesting Hot Pepper Cultured in Greenhouses and Variation in Gene Sequences Encoded in TSWV (시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이)

  • Kim, Chulyoung;Choi, Duyeol;Kang, Jeong Hun;Ahmed, Shabbir;Kil, Eui-Joon;Kwon, Gimyeon;Lee, Gwan-Seok;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.60 no.4
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    • pp.387-401
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    • 2021
  • Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.

Detection of Serum Hepatitis B Virus DNA According to HBV Markers in Chronic Hepatitis B Liver Disease (만성 B형 간질환에서 간염 B virus 표식자 발현에 따른 DNA의 검출)

  • Lee, Dong-Jun;Choi, Jin-Su;Kim, Joon-Hwan;Lee, Heon-Ju
    • Journal of Yeungnam Medical Science
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    • v.14 no.1
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    • pp.155-167
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    • 1997
  • The identification of serum HBV DNA is very important for the assessment of the disease activity in persistent infection, for the evaluation of the infectivity of an individuals blood. The dot blot, however, has limited sensitivity and sometimes inconsistent with other serological markers and clinical settings. Using the most important recent advance in molecular biology, the polymerase chain reaction(PCR), specific DNA sequences can be amplified more than a million-fold in a few hours and with this technique the detection of the extreme low level of DNA is possible. This study was to determine sensitivity of the PCR for the detection of serum HBV DNA in comparison with dot blot analysis and to investigate the serum HBV DNA status and clinical significance of PCR in patients with chronic HBsAg positive liver disease. The subjects of this study were 17 patients with asymptomatic HBsAg carriers(9 HBeAg positive patients, 8 anti-HBe positive patients), 91 chronic hepatitis B(50 HBeAg positive patients, 41 anti-HBe positive patients), 57 liver cirrhosis(21 HBeAg positive patients, 36 anti-HBe positive patients), 27 hepatocellular carcinoma(10 HBeAg positive patients, 17 anti-HBe positive patients). The results were summerized as following; The detection rates of HBV DNA by dot blot, PCR were 58.9%, 72.2% in HBeAg positive patients, 34.3%, 53.9% in anti-HBe positive patients. The detection rates of HBV DNA by PCR in HBeAg negative patients were 25.0% in asymptomatic HBsAg carriers, 61.0% in chronic hepatitis B, 52.8% in liver cirrhosis, 52.9% in hepatocellular carcinoma. The positive rate for HBV DNA is a significant difference between HBeAg positive and negative asymptomatic HBsAg carriers, but not significantly difference in other groups. In conclusions, this study confirmed that the PCR is much more sensitive than the dot blot analysis in detecting the HBV DNA in the sera of patients with chronic liver disease. The presence of HBV DNA in the serum was detected by PCR with higher sensitivity and it suggested that active viral replication is still going on in most patients with chronic HBsAg positive liver disease irrespective of HBeAg/anti-HBe status, and PCR may be used as a prognostic factor in asymptomatic HBsAg carriers.

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Effects of Baicalin on Gene Expression Profiles during Adipogenesis of 3T3-L1 Cells (3T3-L1 세포의 지방세포형성과정에서 Baicalin에 의한 유전자 발현 프로파일 분석)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Chung, Sang-In;Cho, Soo-Hyun;Yoon, Yoo-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.1
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    • pp.54-63
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    • 2010
  • Baicalin, a flavonoid, was shown to have diverse effects such as anti-inflammatory, anti-cancer, anti-viral, anti-bacterial and others. Recently, we found that the baicalin inhibits adipogenesis through the modulations of anti-adipogenic and pro-adipogenic factors of the adipogenesis pathway. In the present study, we further characterized the molecular mechanism of the anti-adipogenic effect of baicalin using microarray technology. Microarray analyses were conducted to analyze the gene expression profiles during the differentiation time course (0 day, 2 day, 4 day and 7 day) in 3T3-L1 cells with or without baicalin treatment. We identified a total of 3972 genes of which expressions were changed more than 2 fold. These 3972 genes were further analyzed using hierarchical clustering analysis, resulting in 20 clusters. Four clusters among 20 showed clearly up-regulated expression patterns (cluster 8 and cluster 10) or clearly down-regulated expression patterns (cluster 12 and cluster 14) by baicalin treatment for over-all differentiation period. The cluster 8 and cluster 10 included many genes which enhance cell proliferation or inhibit adipogenesis. On the other hand, the cluster 12 and cluster 14 included many genes which are related with proliferation inhibition, cell cycle arrest, cell growth suppression or adipogenesis induction. In conclusion, these data provide detailed information on the molecular mechanism of baicalin-induced inhibition of adipogenesis.

Apoptosis-Induced Effects of Extract from Artemisia annua Linné by Modulating Akt/mTOR/GSK-3β Signal Pathway in AGS Human Gastric Carcinoma Cells (AGS 인체 위암 세포에서 Akt/mTOR/GSK-3β 신호경로 조절을 통한 개똥쑥 추출물의 Apoptosis 유도 효과)

  • Kim, Eun Ji;Kim, Guen Tae;Kim, Bo Min;Lim, Eun Gyeong;Kim, Sang-Yong;Kim, Young Min
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.9
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    • pp.1257-1264
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    • 2016
  • Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.

Expression of SARS-3CL Protease in a Cell-Free Protein Synthesis System (무세포 단백질 합성법을 이용한 활성형 SARS-3CL protease의 발현)

  • Park, Sun-Joo;Kim, Yong-Tae
    • Journal of Life Science
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    • v.22 no.4
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    • pp.552-558
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    • 2012
  • Severe acute respiratory syndrome (SARS) is a severe respiratory infectious disease caused by a novel human coronavirus, SARS-CoV. The 3CL protease is a key enzyme in the proteolytic processing of replicase polyprotein precursors, pp1a and pp1ab, which mediate all the functions required for viral genomic replication and transcription. Therefore, this enzyme is a target for the development of chemotherapeutic agents against SARS. A large quantity of active SARS-3CL protease is required for development of anti-SARS agents. Here we have constructed overexpression vector for the production of the SARS-3CL protease. The gene encoding SARS-3CL protease was amplified using polymerase chain reaction and cloned into the pET29a expression vector, resulting in pET29a/SARS-3CLP. Recombinant SARS-3CL protease was successfully synthesized by the dialysis mode of the cell-free protein expression system, and purified by three-step fast protein liquid chromatography using HighQ and MonoP column chromatographies and Sephacryl S-300 gel filtration. In addition, the produced SARS-3CL protease was found to be an active mature form. This study provides efficient methods not only for the development of anti-SARS materials from natural sources, but also for the study of basic properties of the SARS-3CL protease.

Development of Monoclonal Antibody to Cytomegalovirus Maior Immediate-early(α) Protein (거대세포바이러스 조기항원 단백질에 대한 단세포군 항체 제조에 관한 연구)

  • Park, Ji Ho;Sohn, Young Mo;Park, Kyu Hyun;Lee, Won Young
    • Pediatric Infection and Vaccine
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    • v.6 no.2
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    • pp.253-260
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    • 1999
  • Purpose : The purpose of this study is to make and use monoclonal Ab which reacts with CMV major immediate early(${\alpha}$) protein(p72). Methods : Normal human fibroblast(Foreskin derived) was cultured in Eagle's minimal essential medium(MEM) containing 10% cowfetus serum and mouse chondroblast was cultured in P3X63 Ag8.653(ATCC. Maryland USA) to maintain $5{\times}10^5/ml$ cell counts. CMV(KJHJ90) from congenital CMV infected infant's urine was multiplied and used for Ab making. CMV Ag was injected 4 times, 1 week interval into the peritoneal space of 6~8 weeks old mice. And then lymphocyles and fibroblasts taken from spleen were obtained and conjugated. After the conjugated cell cultured, we chose the cell that had high Ab titer using indirect immunofluerescent method. Results : Among the 28 monoclonal antibodies obtained LPC12 and LPC23 reacted highly with nucleus of AD169 infected cell. Purified AD169 after SDS-PAGE, molecular weight of Ag, which reacted with purified monoclonal Ab, was obtained using Western blotting. Monoclonal Ab of LPC12 and LPC23 clone reacted most highly with 72 kd Ag. Conclusion : LPC12 and LPC23 clonal Ab with AD 169(P63-27) is useful on early diagnosis of CMV infection.

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