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Establishment of canine kidney cell line for canine distemper virus replication
황규계,전경익 충북대학교 동물의학연구소 2015 Journal of Biomedical and Translational Research Vol.16 No.1
Kidney cells of canine embryos were separated into single cells using collagenase and dispase. Primary culture was conducted using these cells. To remove fibroblasts, these cells were treated with edetate disodium dihydrate (Na2EDDA), and pure epithelial cells were separated. Recombinant retrovirus particles that manifest teromerase were produced and inoculated into primary culture cells to produce immortalized canine cell strains (JNUCK-1 and JNUCK-2). To examine the characteristics of the produced cell strains, the growth curve, maximum cultured households, and expressed proteins (keratin) were identified. The JNUCK-1 and JNUCK-2 cell lines showed division ability until the 30th generation without growth retardation. JNUCK-1 and JNUCK-2 cell lines clearly expressed telomerase until the 25th generation. The canine distemper virus (CDV) was inoculated into the JNUCK-1 and JNUCK-2 cell lines, as well as in the Madin–Darby canine kidney (MDCK) cell line. The maximum titer of CDV from the JNUCK-1 cell strain was about 200 times higher than that from the MDCK cell strain. However, the JNUCK-2 cell strain produced a lower titer than the MDCK cell strain. We established a new canine kidney epithelial cell line (JNUCK-1) that could produce CDV with high titer.
황규계,전복환,박호선,박송용,김경호,문홍모 대한바이러스학회 1992 Journal of Bacteriology and Virology Vol.22 No.2
Varicella-zoster viruses(VZV) were isolated from the vesicles of the children of typical chicken pox in Korea. These viruses were passed many times in human embryonic lung cells and guinea pig embryonic lung cells. After having been passed many times, these viruses were tested for available biologic and biophysical markers of the attenuated strains in compared with wild strains of VZV. Infectivity ratio of the attenuated strains between guinea pig embryo lung cells and human embryo lung cells was always higher than that of wild strains. This result correlates with the passage history of the attenuated strains in guinea pig embryo lung cells. The attenuated viruses were temperature sensitive at 39C unlike the wild viruses. We, however, could not find any differences between wild strains and attenuated strains on restriction fragment length polymorphism(RFLP) pattern.
국내에서 분리한 수두바이러스의 Restriction Fragment Length Polymorphism분석
황규계,박송용,김성진,유연우,김경호 대한바이러스학회 1991 Journal of Bacteriology and Virology Vol.21 No.2
Varicella-zoster viruses(VZV) were isolated from vesicle fluid of seven korean patients who were 1 to 4 years old and propagated in human embryonic lung fibroblast. We classify the seven VZV isolates in 3 groups by restriction fragment length polyrnorphism(RFLP) analysis using restriction endonucleases-Bam HI, Bgl I, Sma I and Pst I . These 3 types of VZV isolates were different from Ellen strain and Webster strain obtained from ATCC. Compared with Oka strain, two types of them showed different RFLP pattern but the other type showed the same RFLP pattern with above four restriction endonucleases.
박송용,황규계,최문기,유연우,백승복,김경호 대한바이러스학회 1991 Journal of Bacteriology and Virology Vol.21 No.1
A human embryonic lung fibroblast cell line has been established to isolate and propagate varicella-zoster virus (VZV) from Korean patients. The cells were confirmed to be free from extraneous agents as well as mycoplasma. Chromosome analysis of the established cell line revealed that it is less variable than the control cell lines such as WI-38 and MRC-5 obstained from ATCC. From every vesicle fluids of seven varicella patients who were 1 to 4 years old, viruses could be isolated. After the propagation of the viruses, they were proved to be VZV by CPE, IFA, western blot analysis, and electron microscopic photographs. In addition, culture conditions for the multiplication of VZV isolated from Korean patients were established.
제주지역 모기의 계절적 발생소장 및 Real Time RT-PCR을 이용한 Flavivirus 감염조사(2017)
이제욱,황규계,Lee, Che-Wook,Hwang, Kyu-Kye 한국응용곤충학회 2018 한국응용곤충학회지 Vol.57 No.3
기후온난화 현상이 지속되고 있는 제주지역에서 환경적으로 다른 지역 모기의 계절적 발생밀도를 조사하기 위해 제주시의 국제공항, 항만구역과 축사 그리고 서귀포 도심지의 11지점을 선정하여 3월부터 11월까지 매달 2회씩 Black light trap과 BG sentinel trap을 이용하여 모기를 채집하였다. 채집된 모기는 5속 7종. 6,042마리였으며, 이 중 빨간집모기(Culex pipiens)가 4,159마리(68.8%)로 우점종이었으며 흰줄숲모기(Aedes albopictus)는 1,348마리(24.4%)였다. Black light trap를 이용한 채집에서 중앙동주민센터는 트랩당 72.8마리를 채집하여 모기 밀도가 가장 높게 나타났으며 제주국제공항은 트랩당 1.4마리로 가장 낮게 나타났다. BG sentinel trap을 이용한 채집에서는 항만에서 트랩당 71.7마리로 가장 많았고 도심지의 걸매생태공원에서 28.3마리로 가장 낮았다. 시기별로 모기의 밀도는 5월부터 증가하기 시작하여 8월에 1,156마리(19.1%)로 가장 높은 밀도를 나타내었다. 채집된 암컷모기를 종별, 시기별, 지점별로 나누어 pool당 50마리 이하로 설정하여 총 364 pools에서 flavivirus 존재여부를 real time RT-PCR로 검사하였으나, 검출되지 않았다. This study examined the seasonal, regional distribution of mosquito vectors related with disease vectors in the Jeju. From March to November, sample were collected from 11 points in four environmentally different sites in Jeju Island. Samples were collected twice a month using a black-light trap and a BG sentinel trap. Overall, five genera and seven species types of 6,042 female mosquitos were collected. Among the collected mosquitos, 4,159 (68.8%) and 1,348 (24.4%) were Culex pipiens and Aedes albopictus, respectively, making them the dominant species. Additionally, collection using the black light trap produced 72.8 entities per trap in Jungang-dong service center in the center of the city, which was the highest value, while the lowest amount of 1.4 per trap was recovered from the airport. When the BG sentinel trap was used, the largest recovery was observed in the port, where there were 71.7 entities per trap, while the lowest amount of 28.3 entities per trap was recovered at Gealmae Eco Park. The overall number of mosquitoes collected started to increase from May, and reached the largest value of 1,156 (19.1%) in August. Trapped mosquitoes are created 364 pools of up to 50 grains per pool, by season, by environmental, and by species. When the pools were used, no flaviviral infection was observed upon real time RT-PCR.
사람과 기니픽 세포주들에 대한 한탄바이러스의 감수성 및 증식
박호선,황규계,전복환,김혜숙,박송용 대한바이러스학회 1993 Journal of Bacteriology and Virology Vol.23 No.1
Hantaan virus(ROK-84-105) was propagated on various normal human diploid cell lines and guinea pig embryonic lung(GEL) cells. Viral antigens were detected by immunofluorescence assay in each cell lines snd immunoblotting pattern of viral proteins in these cells was identical with that of in Vero E6 cells. So, we found that Hantaan virus was susceptible to not only Vero E6 cells and A549 cell but also normal human diploid cell lines and GEL cells. Contiuous propagation of Hantaan virus in human diploid cell lines and GEL cells was possible and yield of the virus in human embryonic lung fibroblast(LuMA) was enough to rnake a vaccine.
Incidence of Wolf Teeth in Jeju Ponies and Jeju Pony Crossbreds
양재혁,쟈넷한,황규계,임윤규 한국임상수의학회 2015 한국임상수의학회지 Vol.32 No.3
Wolf teeth have been reported with incidence rate of 13% to 31.9% in horses of both sexes. However,there is no report about the incidence of wolf teeth in pony breeds. In order to determine the incidence of wolf teethin Jeju Ponies and Jeju Pony crossbreds, oral examination and palpation of the interdental space of ponies at the JejuRace Park were performed from 2006 to 2010. Results of this study showed that a total of 90 (3.4%) of the 2,675examined Jeju Ponies and Jeju Pony crossbreds had wolf teeth. Among 30 Jeju Ponies that had wolf teeth, 22 weremale (73.3%) and 8 were female (26.7%). Among 60 Jeju Pony crossbreds that had wolf teeth, 25 were male (41.7%)and 35 were female (58.3%). In Jeju Ponies, wolf teeth were most common in 3-years-old, followed by ≥ 4-yearsold,then 2-years-old. For Jeju Pony crossbreds, wolf teeth were most common in 3-years-old, followed by 2-yearsold,then ≥ 4-years-old. In conclusion, the incidence of wolf teeth in Jeju Ponies and Jeju Pony crossbreds wasconsiderably lower than what has been reported for horses in other countries.