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이존화,Nam-Soo Kim,전병우,류시윤,손화영,신현진,박지연,김현철,허진,조정곤,박배근 한국임상수의학회 2010 한국임상수의학회지 Vol.27 No.1
Enterocytozoon bieneusi, a microsporidian species, has emerged as an opportunistic pathogen in AIDS patients. This organism has also been identified in a wide range of animals, and the zoonotic potential of human infections is of particular interest. This study revealed that this organism was found with relatively high prevalence in feces of asymptomatic cattle in Korea. Fecal specimens were obtained from a total of 1,720 cattle in a slaughterhouse located in Chungnam province, Daejeon city and Chonbuk province. After removal of fecal debris by sieving and density gradient centrifugation, samples were examined by microscopic examination and then nested polymerase chain reaction (PCR). Microscopic examination with the modified trichrome staining for the fecal specimens revealed 194(11.28%) positive calves for microsporidia spore. PCR using the specific primer for E. bieneusi revealed 79 (4.59%) positive calves. The infection ratio of microsporidia was higher in March than other season.
이존화 대한면역학회 2015 Immune Network Vol.15 No.1
In the present study, we investigated the protection conferredby a live attenuated Salmonella enterica serovarTyphimurium (ST) strain against Salmonella Typhimurium,Salmonella Gallinarum (SG), and Salmonella Enteritidis(SE) infection in layer chickens. Birds were orally primed withthe attenuated ST strain at 7 days of age and then boostedat 4 weeks post prime immunization (PPI). Sequential monitoringof plasma IgG and mucosal secretory IgA (sIgA) levelsrevealed that inoculation with ST induced a significant antibodyresponse to antigens against ST, SE, and SG. Moreover, significant lymphoproliferative responses to the 3Salmonella serovars were observed in the immunized group. We also investigated protection against virulent ST, SE, andSG strain challenge. Upon virulent SG challenge, the immunizedgroup showed significantly reduced mortality comparedto the non-immunized group. The reduced persistenceof the virulent ST and SE challenge strains in the liver,spleen, and cecal tissues of the immunized group suggeststhat immunization with the attenuated ST strain may not onlyprotect against ST infection but can also confer cross protectionagainst SE and SG infection.
이존화,백병걸,강창원,Lee, John-Hwa,Baek, Byeong-Kirl,Kang, Chang-Won The Korean Society of Veterinary Science 2002 大韓獸醫學會誌 Vol.42 No.4
대장균 K99 섬모의 생합성은 8개로 구성된 K99의 특이 유전자의 발현과 숙주유래 인자에 의해 조절되는 다른 유전자들의 발현에 의존된다. 본 연구에서는 K99섬모 유전자군중 제 3지역 발현에 유전조절자의 관련성 여부를 연구하였다. Gel retardation 분석 방법올 통하여 제3지역의 발현에 관련된 유전조절단위를 함유한 fanF 지역의 단백질 인자가 부착됨을 암시하였다. 이 분석방법을 이용한 결과는 또한 이 단백질 인자가 K99 유전자에서 유래되지 않고 대장균 염색체에서 유래됨을 지적하였다. 이를 보다 더 조사하기 위하여 대장균 염색체에 Tn10 transposon 유전자 변이 실험을 수행하였다. K99 유전자군으로부터 제 1지역과 제2지역의 유전자를 제거시키고, 제 3지역의 유전자인 fanG에 transposon TnlacZ를 삽입한 pTL65-1 plasmid을 제작하였다. 이 pTL65-1는 다시 Tn10으로 염색체가 변이된 대장균에 주입하였다. 3개의 pTL65-1 주입된 Tn10 대장균 변이체 내에서 fanG의 발현이 증가되었다. 이들 변이대장균으로부터 Tn10이 어떤 염색체 유전자 부위를 변이 시켰는지 확인하기 위해서 변이부위 유전자를 cloning하여 염기서열을 분석하였다. 이중 2개의 clone이 동일하였으며 지금까지 알려지지 않은 유전자였다. 이들 2개의 변이체 내에서 fanG의 발현은 대조군과 비교해 약 4.2배 증가 되였다. 결론적으로 이들 2개의 clone으로부터 유래된 인자는 지금까지 알려지지 않은 제 3지역의 억제 조절자임을 나타내었다.
The Effect of Seminal Plasma on Chilling and Freezing of Canine Spermatozoa
유명조,이존화,김인식,박진호,권중기,김종훈,유일정,김범석 한국임상수의학회 2007 한국임상수의학회지 Vol.24 No.4
Seminal plasma (SP) is usually removed from semen that is to be cryopreserved. However, some reports indicate that SP has beneficial effects on spermatozoa during chilling and freezing. The purpose of this study was to determine the effect of SP on sperm survival by adding SP to the extender before cooling and freezing canine spermatozoa. In replicate experiments, ejaculates obtained from four healthy dogs (1-4 years old) of various breeds were pooled, centrifuged at 300 × g for 10 min at 25oC, and the supernatant of seminal plasma was decanted. Spermatozoa were suspended in egg yolk-Tris (EYT) buffer. The study comprised two experiments: [Exp 1] Sperm were suspended in EYT extender containing either 0, 20, 40, 80 or 100% SP and were slowly cooled to 4oC for 2 h or held at 25 oC as controls. Sperm concentration was adjusted to 2 × 108/ml. [Exp II] Sperm samples, each of which contained 1 × 108/ml, were assigned to nine groups to be frozen. In the first four groups, sperm in EYT containing either 20, 40, 80 or 100% SP were cooled to 4oC, then diluted to contain final concentrations of EYT + 0.6 M glycerol and then were frozen. The final concentrations of SP were 10, 20, 40 or 50%. In the other four groups, sperm in EYT alone were first cooled slowly to 4oC, then diluted to contain final concentrations of EYT + 0.6 M glycerol plus 10, 20, 40 or 50% SP and then were frozen. Spermatozoa, which chilled in EYT alone and diluted to contain final concentrations of EYT + 0.6 M glycerol without seminal plasma, and then frozen, was regarded as control. Spermatozoa were frozen at 25oC/min of cooling rate in plastic straws that were suspended above liquid nitrogen and thawed in water at 38oC for 1 min. Sperm survival was assayed by determining progressive motility and integrity of plasma and acrosome membranes. Progressive motility was determined by microscopic examination at 200 × magnification. Membrane integrity was assessed by use of a double fluorescent dye, and acrosome integrity by staining sperm with Pisum sativum agglutinin. The results of the first experiment showed that adding SP did not improve motility of spermatozoa compared to those incubated without SP regardless of temperature. The results of the second experiment showed that spermatozoa suspended in EYT+0.6 M glycerol containing SP exhibited the higher progressive motility before being frozen (P < 0.05). However, frozen-thawed spermatozoa that had suspended in EYT + 0.6 M glycerol containing SP showed the similar or lower viability (P < 0.05). In summary, although seminal plasma did not affect spermatozoa that were chilled in EYT without cryoprotectant (CPA), addition of seminal plasma to EYT containing CPA did significantly improved progressive motility of canine spermatozoa that were chilled.
PC12 세포에서 신경전달물질 방출을 저해하는 생리활성물질 FS11052의 탐색
이윤식,이존화,Lee, Yun-Sik,Lee, John Hwa 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.2
We established an in vitro experimental system using the following procedure. We first introduced tritium-labeled norepinephrine ([$^3$H]-NE) into PC12 cells, The [$^3$H]-NE incorporated into PC12 cells were then stimulated by a high concentration (60 mM) of $K^+$ buffer during 12 minutes. Then, we collected $100{\mu}l$ supernatant and counted the amount of [$^3$H]-NE release from PC12 cells with a scintillation counter. After screening fungal, Streptomyces spp. or bacterial product using this experimental sytem, we obtained FS11052 from Streptomyces spp. which inhibited [$^3$H]-NE release from PC12 cells. FS11052 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons, The inhibitory effect was seen even when the PC12 cells were treated with low $K^-$ buffer containing ionomycin ($1{\mu}M$) as an ionopore. This result suggests that the inhibitory action of FS11052 on neurotransmitter release appeared after the influx of $Ca^{2+}$.
Jin, Song Jun,Lee, John Hwa,Kang, Chang Won 한국수의공중보건학회 2001 예방수의학회지 Vol.25 No.1
한우의 골 관절염에서 IGF-I/IGFBPs axis의 변동을 연구하기 위하여 활액 내 IGF-I의 농도, IGFBPs의 patterns 및 IGF-I과 IGFBP-3의 protease activity를 정상 한우 혈청과 비교 분석하여 다음과 같은 결과를 얻었다. 한우 골관절염 활액 내 총 IGF-I의 농도는 정상 활액 보다 증가하였고 free IGF-I의 농도는 정상 활액 보다 감소하였다.(n = 30, p < 0.05). 한우 골 관절염 활액 내 IGF-I proteolytic activity는 정상 활액 보다 활성화되었다. Western ligand blotting의한 IGFBPs의 partterns에서 정상 활액과 골 관절염 활액에서 IGFBP-3, -2, -1 및 -4을 관찰하였다. 이 중 IGFBP-2는 골 관절염 활액에서 정상 활액보다 증가하였으며, Western immunoblotting에서의 IGFBP-3는 골 관절염 활액에서 정상 활액보다 증가하였다. 정상 활액 내 IGFBP-3 proteolytic activity는 정상 혈청과 골 관절염 활액보다 활성이 증가하였으며, 이는 EDTA에 의하여 차단되었다. 정상 활액과 골 관절염 활액 내 IGF-I의 150 kDa과 50 kDa carrier proteins는 유사하였다. 따라서 한우 골 관절염 활액 내 IGF-I/IGFBPs axis 변화는 골 관절염에 관여할 것으로 사료된다.