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Improvement of Pregnancy Rate in Preimplantation Genetic Diagnosis with FISH Procedure by the Laboratory Optimization and Experiences (형광직접보합법을 이용한 착상전 유전진단 기법의 최적화와 경험 축적에 의한 임신율의 향상)

  • Lim, Chun-Kyu;Min, Dong-Mi;Lee, Hyoung-Song;Byun, Hye-Kyung;Park, So-Yeon;Ryu, Hyun-Mee;Kim, Jin-Young;Koong, Mi-Kyoung;Kang, Inn-Soo;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.29-39
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    • 2004
  • Objectives: This study was performed to evaluate the laboratory system for successful PGD using fluorescence in situ hybridization (FISH) and the clinical outcome of PGD cycles in five years experiences. Methods: A total of 181 PGD-FISH cycles of 106 couples were performed, and diagnosed chromosome normality in the preimplantation embryos. The laboratory and clinical data were classified by the following optimization steps, and statistically analyzed. Phase I: Blastomere biopsy with two kinds of pipettes, removal of cytoplasmic proteins without treatment of pepsin and culture of biopsied embryos with single medium; Phase II: Blatomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with single medium; Phase III: Blastomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with sequential media. Results: A total of 3, 209 oocytes were collected, and 83.8% (2, 212/2, 640) of fertilization rate was obtained by ICSI procedure. The successful blastomere biopsies were accomplished in 98.6% (2, 043/2, 071) of embryos, and the successful diagnosis rate of FISH was 94.7% (1, 935/ 2, 043) of blastomeres from overall data. Embryo transfers with normal embryos were conducted in 93.9% (170/181) of started cycles. There was no difference in the successful rate of biopsy and diagnosis among Phase I, II and III. However, the pregnancy rate per embryo transfer of Phase III (38.8%, 26/67) was significantly (p<0.05) higher than those of Phase I (13.9%, 5/36) and Phase II (14.9%, 10/67). Conclusions: The laboratory optimization and experience for the PGD with FISH procedure can increase the pregnancy rate to 38.8% in the human IVF-ET program. Our facility of PGD with FISH provides the great possibility to get a normal pregnancy for the concerned couples by chromosomal aberrations.

Decay Rate and Nutrients Dynamics during Decomposition of Oak Roots (상수리나무 뿌리 분해 및 분해과정에 따른 영양염류 변화)

  • 문형태
    • The Korean Journal of Ecology
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    • v.27 no.3
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    • pp.165-171
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    • 2004
  • Weight loss and nutrients dynamics during decomposition of oak roots (diameter classes: R₁〈0.2㎝, 0.5㎝〈R₂〈1㎝, 1㎝〈R₃〈2㎝, 2㎝.〈R₄〈4㎝) (Quercus acutissima) were studied for 33-months in Kongiu, Korea. After 33-months, decomposition rate of R₁, R₂, R₃ and R₄ was 49.6%, 47.5%, 66.4% and 66.1%, respectively. The decomposition constant(k) for R₁, R₂, R₃, and R₄ was 0.249/yr, 0.234/yr, 0.397/yr and 0.393/yr, respectively. Larger diameter class of the root lost more weight than smaller diameter class. N concentration in decomposing oak roots increased in all diameter classes. After 33-months, remaining N in R₁, R₂, R₃ and R₄ was 66.5%, 80.7%, 84.4% and 44.4%, respectively. K concentration in decomposing oak roots decreased in early part of decomposition and then increased in later stage of decomposition. After 33-months, remaining P in R₁, R₂, R₃ and R₄ was 64.7%, 62.4%, 93.1% and 30.7%, respectively. K concentration in decomposing oak roots decreased rapidly in early stage of decomposition. Remaining K in R₁, R₂, R₃ and R₄ was 11.6%, 10.6%, 5.9% and 7.7%, respectively. Ca concentration in decomposing oak roots showed different among diameter classes. After 33-months, remaining Ca in R₁, R₂, R₃ and R₄ was 66.2%, 51.0%, 39.1% and 48.3%, respectively. Initial concentration of Mg in oak root was higher in smaller diameter class. After 33-months, remaining Mg in R₁, R₂, R₃ and R₄ was 15.3%, 29.9%, 24.5% and 69.4%, respectively.

Effects of electromagnetic stimulation on neurogenesis and neuronal proliferation in rat hippocampal slice culture (실험 쥐 해마조직배양에서 전자기 자극이 신경조직발생 및 증식에 미치는 영향)

  • Kim, Deok-Soo;Choi, Eung Sang;Chae, Soo Ahn
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.558-564
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    • 2006
  • Purpose : Transcranial electromagnetic stimulation(TMS) is a noninvasive method which stimulates the central nervous system through pulsed magnetic fields without direct effect on the neurons. Although the neurobiologic mechanisms of magnetic stimulation are unknown, the effects on the brain are variable according to the diverse stimulation protocols. This study aims to observe the effect of the magnetic stimulation with two different stimulation methods on the cultured hippocampal slices. Methods : We obtained brains from 8-days-old Spague-Dawley rats and dissected the hippocampal tissue under the microscope. Then we chopped the tissue into 450 µm thickness slices and cultured the hippocampal tissue by Stoppini's method. We divided the inserts, which contained five healthy cultured hippocampal slices respectively, into magnetic stimulation groups and a control group. To compare the different effects according to the frequency of magnetic stimulation, stimulation was done every three days from five days in vitro at 0.67 Hz in the low stimulation group and at 50 Hz in the high stimulation group. After N-methyl-D-aspartate exposure to the hippocampal slices at 14 days in vitro, magnetic stimulation was done every three days in one and was not done in another group. To evaluate the neuronal activity after magnetic stimulation, the $NeuN/{\beta}$-actin ratio was calculated after western blotting in each group. Results : The expression of NeuN in the magnetic stimulation group was stronger than that of the control group, especially in the high frequency stimulation group. After N-methyl-D-aspartate exposure to hippocampal slices, the expression of NeuN in the magnetic stimulation group was similar to that of the control group, whereas the expression in the magnetic non-stimulation group was lower than that of the control group. Conclusion : We suggest that magnetic stimulation increases the neuronal activity in cultured hippocamal slices, in proportion to the stimulating frequency, and has a neuroprotective effect on neuronal damage.

Evaluation of Cabbage- and Broccoli-genetic Resources for Resistance to Clubroot and Fusarium Wilt (뿌리혹병 및 시들음병에 대한 저항성 양배추와 브로콜리 유전자원 탐색)

  • Lee, Ji Hyun;Jo, Eun Ju;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Research in Plant Disease
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    • v.20 no.4
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    • pp.235-244
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    • 2014
  • Clubroot and Fusarium wilt of cole crops (Brassica oleracea L.) are destructive diseases which for many years has brought a decline in quality and large losses in yields all over the world. The breeding of resistant cultivars is an effective approach to reduce the use of chemical fungicides and minimize crop losses. This study was conducted to evaluate the resistance of 60 cabbage (B. oleracea var. capitata) and 6 broccoli (B. oleracea var. italica) lines provided by The RDA-Genebank Information Center to clubroot and Fusarium wilt. To investigate resistance to clubroot, seedlings of the genetic resources were inoculated with Plasmodiophora brassicae by drenching the roots with a mixed spore suspension (1 : 1) of two isolates. Of the tested genetic resources, four cabbage lines were moderately resistant and 'K166220' represented the highest resistance to P. brassicae. The others were susceptible to clubroot. On the other hand, to select resistant plants to Fusarium wilt, the genetic resources were inoculated with Fusarium oxysporum f. sp. conglutinans by dipping the roots in spore suspension of the fungus. Among them, 17 cabbage and 5 broccoli lines were resistant, 16 cabbage lines were moderately resistant, and the others were susceptible to Fusarium wilt. Especially, three cabbage ('IT227115', 'K161791', 'K173350') and two broccoli ('IT227100', 'IT227099') lines were highly resistant to the fungus. We suggest that the resistant genetic resources can be used as a basic material for resistant B. oleracea breeding system against clubroot and Fusarium wilt.

Effect of Dietary Fish Oil on Lipid Peroxidation in Rats Liver and Brain During Postnatal Development (어유섭취가 출생후 발달과정의 흰 쥐의 간과 뇌조직의 지질과산화와 그 관련기능에 미치는 영향)

  • 박명희
    • Journal of Nutrition and Health
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    • v.20 no.2
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    • pp.111-121
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    • 1987
  • Lipie peroxide formation, antiperoxidative s system and body adaptability for handling lipid p peroxide were examined in the first and second g generations of rats fed fish oil. Mackerel oil(MO) was used and four other dietary oils and fat, i.e. soybean oil(SO), perilla oil(PO), rapeseed oil(RO) and beef tallow(BT) were also employed to compare the effect of fish oil. Synthetic diets containing these five dietary fats at the level of 1O%(w/w), were given to the correspond­m ing groups of male and female rats weighing about 70 grams. After 34 days of feeding, male a and female rats were mated and their offsprings were raised throughout suckling (17, 26 days) and weanling (39 days) periods. Liver lipid pero­x xide level was highest in MO group of both first (mother rats after lactation) and second genera­t tions of 17 and 26 days old, but not of 39 days old. During suckling period, liver lipid peroxide level was well matched to total unsaturation of dietary fat. Brain lipid peroxide levels were not different among five groups. Liver $alpha$-tocopherol a and reduced glutathione (GSH) levels were lowest in MO fed first generation. In second generation, $alpha$-tocopherol level was also low in MO group, although the effect was less pronoun­c ced, but GSH level was not different from other groups. Oxidized glutathione (GSSG) level did not consistently vary by change in dietary fat. Glutathione peroxidase activity increased as young rats grew up to 39 days. Superoxide d dismutase activity change was insignificant by a age, but was shown as lowest in MO group. At the age of 26 and 39 days, liver glutatione peroxidase activity was increased as was level of lipid peroxide, suggesting that this is the one of the mechanisms responsible for body adapta­b bility for protection against the accumulation of lipid peroxide.

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Effect of Different Soil Water Potentials on Growth Properties of Northern-Highbush Blueberry (토양수분포텐셜이 북부형 하이부쉬 블루베리의 생육에 미치는 영향)

  • Kim, Hong-Lim;Kwack, Yong-Bum;Kim, Hyoung-Deug;Kim, Jin-Gook;Choi, Young-Hah
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.2
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    • pp.161-167
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    • 2011
  • The soil moisture has an important effect on growth and development of highbush blueberry (HB), mainly because the root system, devoid of root hairs, is superficial. Moreover, the texture and organic matter content of Korean soil is different from the main producing counties, such as USA and Canada. To facilitate the growth and development of HB and long-term maintenance of productivity, the research related to soil moisture condition in Korea should be the priority. This study was performed to investigate the growth properties of the HB in various soil moisture conditions in order to determine the irrigation trigger point and optimum soil water potential. The texture of soil used in this experiment was loam. For the experiments, the soil was mixed with peatmoss at a rates 30% (v/v). Irrigation was scheduled at -3, -4, -5, -8, -15 and -22 kPa soil water potential then investigated leaf macronutrient, bush growth, and fruit properties. The leaf K content of HB showed the same trend in the soil water potential, but Leaf P and Mg content was highest in -5 and -22 kPa, respectively. The productivity and growth amount of HB showed the peak at the range of -4~-8 kPa as normal distribution pattern, and greatly decreased at above -15 kPa. Total dry weight and Cane diameter were highest at -4 kPa, plant width, fruit weight and yield were highest at -5 kPa, and plant height, cane number and shoot tension were highest at -8 kPa. Soluble solids content showed same trend in the soil water potential, but titratable acidity, anthocyanins and total polyphenols were not significantly different. Therefore, the optimal soil water potential for the development and a maximum production of HB were a range of -4~-8 kPa, and the recommended ideal irrigation trigger point was within -15 kPa.

Correlation between Steroid Hormone Metabolites and Leiomyomas of Uterus (자궁근종과 성호르몬 대사물과의 연관성)

  • Bai, Sang-Wook;Jung, Byung-Hwa;Chung, Bong-Chul;Jeon, Jin-Dong;Lee, Hyun-Jung;Kwon, Han-Sung;Chung, Kyung-Ah;Kim, Sei-Kwang;Park, Ki-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.4
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    • pp.279-286
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    • 2001
  • Objective: To elucidate 1) whether there are any differences in the urine concentrations of steroid hormone metabolites between patients with leiomyoma and normal controls 2) the correlation between urinary profiles of steroid hormones and leiomyomas of the uterus according to their type, location, volume, and weight. Materials of Methods : The study population consisted of 37 premenopausal patients with uterine leiomyoma and the control group consisted of 25 premenopausal normal volunteer women without uterine leiomyoma. Confirmation of the existence of uterine leiomyoma was done by ultrasonography and histopathological examination after surgery. The volume of the leiomyoma was estimated by trans-abdominal and/or trans-vaginal ultrasonography. The Leiomyomas were divided into 3 types (subserosal, intramural and submucosal). Seventeen patients had subserosal type of leiomyoma, 10 with the intramural type and 10 with the submucosal type. The locations of the leiomyoma were also divided into 3 groups (fundus, body and isthmus). Seventeen patients showed a fundus location, 10 in body, and 10 in isthmus. We compared urinary profiles of the endogenous steroids between patients with leiomyomas and normal controls, and also investigated the relationship between urinary profiles of the endogenous steroids and leiomyomas according to their type, location, volume and weight by using highly sensitive Gas Chromatography-Mass Spectrometry (GC-MS) system. Results: The mean ages of the patients with leiomyomas and the control group were $43.1{\pm}5.6$ and $40.6{\pm}7.2$ years, the weights were $63.4{\pm}7.3$ and $59.4{\pm}8.1\;kg$, and their heights were $155.4{\pm}4.8$ and $159.3{\pm}4.8\;cm$ respectively. Seventeen patients had subserosal, 10 had intramural, and 10 had submucosal leiomyomas. There were 17 patients with leiomyoma located in fundus, 10 in body and 10 in isthmus. $17{\beta}$-estradiol, 5-AT, 11-keto ET, $11{\beta}$-hydroxy An, $11{\beta}$-hydroxy Et, THS, THA, THE, a-cortolone, a-cortol, $\beta$-cortol, $11{\beta}$-OH Et/$11{\beta}$-OH An and E2/E1 were significantly increased in patients with leiomyoma than in the control group. $17{\beta}$-estradiol was significantly increased in the intramural and the submucosal types than in the subserosal type. There was no significant difference in the concentrations of urinary steroids according to the locations of leiomyomas. There was no significant relationship between the concentration of urinary steroids and the volume of the leiomyomas. $17{\beta}$-estradiol significantly decreased as the weight of uterus increased (r=-0.322, p=0.04). Conclusion: The concentrations of steroid hormone metabolites were generally increased in patients with leiomyoma but were not significantly related to the volume and weight of the leiomyomas. Our study suggests that steroid hormones may be involved in the initiation of leiomyomas but may not be involved in their progression. In addition, the concentrations of steroid hormone metabolites are not related to the leiomyoma type and location.

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Index Cases in Pediatric Surgery - a National Survey by the Korean Association of Pediatric Surgeons, 2000 - (소아외과의 지수 질환 - 대한소아외과학회 정회원을 대상으로 한 2000년도 전국 조사 -)

  • Lee, M.D.;Kim, S.Y.;Kim, W.K.;Kim, I.K.;Kim, S.C.;Kim, S.K.;Kim, J.E.;Kim, J.C.;Kim, H.H.;Park, K.W.;Park, W.H.;Seo, J.M.;Song, Y.T.;Oh, S.M.;Yoo, S.Y.;Lee, D.S.;Lee, S.K.;Lee, S.C.;Chung, S.Y.;Chung, S.U.;Jung., E.S.;Jung, P.M.;Cho, M.H.
    • Advances in pediatric surgery
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    • v.7 no.2
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    • pp.147-156
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    • 2001
  • Membership qualifications and recognition of the subspecialty training programs by the Korean Association of Pediatric Surgeons(KAPS) have been determined by criteria based on the number of neonatal and total pediatric surgical cases registered, since 1989 (Index cases). The numbers are based on a survey of the 14 founding members of the society by Jung et al. in 1987. The current survey is to review the present status of the index cases from 1997 to 1999 among 36 members of the KAPS. Two surveys were undertaken for this study. In the first survey, cases were collected by the registration form, composed of neonatal(N), important pediatric surgical cases (I), tumor and other similar operations(T) and other common pediatric surgical cases(O). Thirty members responded. and the result was discussed at the Topic Discussion section of the 15th Annual Congress of KAPS, 2000. The second survey analyzed additive data, proposed during discussion at the Congress. Twenty-three members responded. This report is the analysis of the both surveys. The average numbers of the cases/year/surgeon of N, I, T, O and total were 19.5, 51.8, 9.5, 77.1 and 356.5. respectively. The number of index cases(N+I+T)/year/surgeon was 80.8. The ratio of (N+I+T)/total cases was 0.3. Seventeen of 30 members have more than 20 cases of N per year. Twelve members have more than 150 cases, and 13 do more than 100 cases of I per year. Fourteen members have more than 10 cases of T per year. Nineteen and 10 members experienced more than 150 and 100 of index cases(N+I+T) per year. A new list of the index cases and coding system are proposed for the future regular update.

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Thioredoxin Peroxidase Manifestation in Radiation-Induced White Rat Lung Tissues (방사선 조사후 손상된 백서 폐조직에서의 Thioredoxin Peroxidase의 발현)

  • Chong, Seong-Cheoll;Park, Joon-Seong;Park, Jee-Won;Lee, Sun-Min;Park, Kwang-Joo;Hwang, Sung-Chul;Lee, Yi-Hyeong;Hahn, Myung-Ho;Oh, Young-Taek;Kim, Hyung-Joong
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.5
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    • pp.650-659
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    • 1999
  • Background/Aims: It is well recognized that all aerobic cells have the protective mechanisms in order to minimize the tissue damage induced by various reactive oxygen species(ROS). Thioredoxin peroxidase(TPX) which has been recently identified and characterized functions to convert peroxide to water. The protein is also found in various subtypes(TPX-A & B, MER5, HS22 and HORF-06) and is known to be ubiquitous in most human cells. Especially, ischemic brain injuries, partial hepatectomy and radiation induced DNA damages. In treating lung cancer, radiation therapy has a major place in the local control and the relief of symptoms, but radiation induced free radical injury and resulting pulmonary fibrosis has been the major drawback of the therapy. However, little is known about the protective mechanisms and biologic modulations against radiation-induced tissue damages. Methods: Eighteen mice were divided into six groups, 3 in each group, and fifteen had received 900cGy of radiation. The mice were sacrificed according to the pre determined time schedule; immediate, 1, 2, 3 and 6 weeks after irradiation. Extracts were made from the lungs of each mice, Western blot analysis of various subtypes of TPX were done after SDS-P AGE. Examination of H & E stained slides from the same irradiated specimens and the control specimens were also performed. Results: No difference in the intensity of the immunoreactive bands in the irradiated lung samples of the mice compared to the unirradiated control was observed regardless of the time intervals, although H & E examination of the sample specimens demonstrated progressive fibrotic changes of the irradiated lung samples. Conclusion: In conclusion, according to our data, it is suggested that various thioredoxin peroxidase subtypes and catalase which are known to be increased in many repair processes may not be involved in the repair of the radiation injury to the lung and subsequent fibrosis.

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Epidemiological Studies of Avian Reovirus Infection in Broilers in Korea (국내 육계에서의 조류 레오바이러스 감염에 대한 역학 조사)

  • Kim, J.M.;Kim, M.J.;Song, J.S.;Mo, I.P.
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.85-99
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    • 2008
  • Avian reovirus (ARV) is a causative agent of viral arthritis/tenosynovitis, and malabsorption syndrome in broiler. The characteristics of malabsorption syndrome caused by ARV are diarrhea, poor feed conversion and stunting. Therefore, ARV infection has been recognized as one of the most important disease in the poultry industry because of economical losses. However, few study of ARV infection in broiler industry has been conducted in Korea. To evaluate the presence of ARV infection in broiler farms, epidemiological survey such as serological test and virus isolation has been conducted. For the serological survey using ELISA method, we selected five broiler farms which were located at different area and had a history of growth retardation, lameness, diarrhea and poor feathering. From these farms serum samples were collected at 1 day, 14 days and market age. All these farms had no history of vaccination against ARV. In addition to serological survey, we tried to isolate ARV from birds of designated farms at market age and collected feces and tissue samples such as cecal tonsil, intestine and liver. We were identified ARV by RT-PCR and transmissible electron microscopy. The samples were inoculated into 9-day-old embryonated eggs via the chorioallantoic membrane to observe the pock formation. For the pathogenicity test of ARV isolates, we inoculated with the isolates to the right footpad of 3-week-old SPF chicks and observed clinical signs and pathological changes for 14 days after challenge. Most broilers sampled for serological survey have maternal antibodies which were widely distributed at 1 day and decreased by 14 days. However, at the market age several broiler farms showed fairly high antibody titer against ARV. This increase of antibody titer at market age means the possible infection of ARV during the grow-out period. Among total 15 samples for the isolation of ARV. 2 samples were positive by RT-PCR and finally identified as a ARV. We inoculated these isolates in the SPF birds and observed that the antibody titer was increased from 7 days after challenge. However, we did not find any clinical signs both control and challenge groups. Based on the above results, it is clear that the ARV infection has been circulated in the broiler industry and caused significant economic losses. Further study is needed to evaluate the virulence of the isolates in the digestive system of broiler and the molecular characteristics of isolates.