Development of a Mobile Application, "Wild Flowers of Bukhansan National Park (version 1.0)", for Identification of Plants in Bukhansan National Park

Kim, Sang-Tae,Lee, Seung-Yeon,Kim, Seung-Chul,Byun, Hye-Won,Lee, Sang-Tae,Kim, Mu-Yeal,Hong, Seok-Pyo,Chung, Young-Jae,Park, Ki-Ryong,Lee, Chung-Hee,Lee, Joong-Ku,Heo, Kyeong-In,Lee, Ji-Ye,Lee, Eun-Je National Science Museum of Korea 2011 Journal of Korean nature Vol.4 No.3

We developed the educational purpose mobile application, named "Wild Flowers of Bukhansan National Park (version 1.0)", aiming for easy identification of wild flowers for students and visitors in the park. When visitors find a flower or part of plant in the park, visitors can search for its name utilizing the pictures and characters provided in their own smartphone mobile devices or tablet PCs. The application provides pictures of wild flowers in the park and character-based searching system based on 12 diagnostic features (e.g., growth form, leaf arrangement, flower symmetry, petal color, petal number, sepal number, etc). We adopted the complete floristic survey of Chung and Lee (1962) and added species that we confirmed their distribution in the park during the development of this application. In summary, number of vascular plants in this park was estimated to be 428 taxa including 100 families, 280 genera, 327 species, 1 subspecies, 50 varieties, and 5 formas. We provided a total of 588 pictures representing 358 taxa and each taxon includes multiple pictures in many cases. Included identification quizzes can be an efficient educational tool as well as fun activity for students and visitors who are learning plant species in Korea. Our next step will include GPS function in the application for indicating visitor's location and for providing previously reported sites of the species that we interested in the map of the park. The future application which includes GPS function will be a valuable tool for the monitoring of rare plants, plant researches related to the climate changes, etc. We currently provide Korean iPhone version only, and English version and both of android versions will be serviced soon.

Distribution of the Fairy Pitta (Pitta nympha) in the South Korea: A focus on protected areas

Gil-Pyo Hong(홍길표),Jae-Hoon Kim(김재훈),Seul-Gi Seo(서슬기),So-Yeon Cho(조소연),Bo-Yeon Hwang(황보연),Jong-Hyun Park(박종현),Seung-Yeon Lee(이승연),Ga-Young Kim(김가영),Wan-Hee Nam(남완희),Ha-Cheol Sung(성하철) 한국조류학회 2021 한국조류학회지 Vol.28 No.2

국내 팔색조(Pitta nympha) 분포 현황을 알아보고자 전국 국립공원 등 보호지역 중심으로 총 19개 지역을 조사하였고, 전문가 네트워크를 통해 자료를 보강하였다. 전국 19개 지역을 조사한 결과 북한산, 지리산 등 내륙지역 7개 국립공원에서는 팔색조가 확인되지 않았고, 경남 거제, 남해, 해남, 장흥 고흥 등 남해안과 인접한 12개 지역에서 팔색조 서식이 확인되었다. 또한 전문가 네트워크를 통해 수집한 팔색조 출현 위치 정보 82건을 분석한 결과, 전국 8개 광역지자체 모두 팔색조 서식이 확인되었고, 내륙지역(26.8%)보다 해안지역(73.2%)에서 더 많이 출현하였다. 연구 결과를 종합해보면, 우리나라의 팔색조는 주로 제주도와 남해안 지역을 중심으로 많이 분포하며, 그 보다 적은 개체수가 내륙 지역에 넓게 분포하고 있는 것으로 나타났다. The present study investigated the current distribution of Fairy Pitta (Pitta nympha) in South Korea. A total of 19 study areas, including protected areas such as national and provincial parks in South Korea were surveyed, and additional data obtained from an expert network. According to the results, Fairy Pitta was absent in seven inland national parks, including Mt. Bukhan and Chiri parks. Fairy Pitta inhabited 12 areas near the southern coast of South Korea, including Geoje, Namhae, Haenam, Jangheung, Goheung, and Gyeongnam Province. The analysis results of 82 datasets obtained from experts with regard to the locations where Fairy Pitta have been spotted showed that the bird inhabits all the eight metropolitan regions in South Korea, and is more frequently spotted in coastal areas (73.2%) than in inland areas (26.8%). The findings demonstrate that Fairy Pitta is mainly distributed in Jeju Island and the southern coast of South Korea, and is distributed in inland areas at relatively lower numbers.

돼지 미성숙 난포란의 회수 및 체외수정에 관한 연구

박성원,김정혁,홍승표,이지삼,정장용,박희성 진주산업대학교 농업기술연구소 1999 農業技術硏究所報 Vol.12 No.-

본 연구는 돼지의 체외수정에 있어서 난포란의 회수방법과 체외수정란 및 체외발달율을 조사하여 체외수정의 기본 원리를 밝히고자 본 연구를 수행하였다. 도축장으로부터 채집한 난소의 난포에서 흡입법, 세절법 및 흡입후세절법 등으로 난포란을 채취하여 등급별로 분류하여 회수율을 조사하였으며, 채취한 난포란은 액상정액과 정소상체미부 정액을 사용하여 체외수정을 시킨후 TCM-199 와 NCSU-23 배양액으로 체외배양을 실시하였다. 본 연구에서 얻은 결과를 요약하면 다음과 같다. 회수방법당 각각 37개의 난소에서 회수한 난포란의 수는 흡입법이 1,365개, 세절법이 1,884개 및 흡입후 세절법이 3.830개로써 난소당 난포란의 수는 각각 36.9, 50.9 및 103.5개로써 회수방법간에 유의적(P<0.05)인 차이가 있었다. 액상정액과 정소상체미부 정액을 사용하였을 때 수정율은 각각 83.0 및 83.% 로써 정액간에 유의적(P<0.05) 인 차이는 없었다. 체외수정이 이루어진 난자들의 분할율도 액상정액(60.8%) 과 정소상체미부 정액(69.0%) 간에 유의적(P<0.05) 인 차이는 없었다. 2-4세포기로의 발달율은 24.6 및 28.4%로써 배양액간에 위의적(P<0.05)인 차이는 없었으며, 상실배로의 발달율도 TCM-199 와 NCSU-23 배양액이 각각 14.8 및 24.5% 로써 이들간에 유의적(P<0.05)인 차이는 없었다. To improve the efficiency of in vitro production of embryos with follicular oocytes in pig, the recovery rates, in vitro fertilization and development, and the time required for collectng and processing oocytes by aspiration with or without slicing were evaluated comparatively. The oocytes were collected by aspiration of follicles(2-6mm) with or without slicing ovaries after aspiration, and classified into grade I-IV oocytes by the morpholoy of cumulus cells attached and the homogeneity of cytoplasmic granules. The grade Ⅰ-Ⅱ oocytes were matured invitro for 46 hours in TCM-199 and NCSU-23 supplemented with 1㎍/㎖ FSH, 2IU/㎖ hCG, 1㎍/㎖ estradiol-17 at 39℃ under 5% CO_2 in air. They were fertilized in vitro by epididymis sperm and ejaculated sperm treated with hypotaurin for 24 hours. and then the zygotes were co-cultured in vitro with pFF(porcine follicular fluid) for 7-9 days. The results obtained were as follows: The number of ocytes recovered per ovary was averaged 30.7 by aspiration, 50.8 by slicing and 103.5 by aspiration post slicing. The number of grade Ⅰ oocytes recovered was total 0.2% by aspiration, 0.1% by slicing and 0.1% by aspiration post slicing. The oocytes recovered per ovary was significantly(P<0.05) higher as recovery methods. The cleavage rates of the embryos obtained from 83.0(ejaculate sperm) and 83.1(epididymis sperm) respectively(P<0.05). In vitro developmental rates(9.9 and 18.6%) of embryos cultured in TCM-199 and NCSU-23 up to morula stage were non significantly(P<0.05) different between the culture media.

體驗的 自我의 崩壞와 故鄕喪失의 詩的 形象化

朴昇杓 동의대학교 인문과학연구소 1996 인간과 문화 연구 Vol.1 No.-

Diese Arbeit will ein Gesamtlild der Lyrik des 19. Jahrhunderts entwerfen und zuerst einige ausgewahlte von Morike, der Droste-Hulshoff betrachten. In Morikes Dicdtung finden wirtrotz aller individueller Eigenart wesentliche Voraussetzungen und Entwichklungaszuge der Lyrik in den beiden drsten Dertteln des 19. Jahrhunderts. Die neuen Ansatze von Morikes Lyrik scheinen uns in der Dichtung der Droste entwichkelt. Die lyrichen Moglichkeiten der Zweiten Dichter erscheinen beschrankter als die Mortkes. Die bedeutenden Gedichte der Droste sind in einem Zeitraum von zehn Jahren nststanden und zeigen geringe Wandlungen, nur der Zwiespalt der Zeit pragt sich in ihr aus. Die beide Dichter stehen auf der Schwelle der Zeit, zwischen den Epochen. So entspricht die Entwichlungsstufe ihrer Gedichte ihrem geistes geschichtlichen Standort. Im Gegensatz zur Nacht der Romantik steht der Realismus des 19. Jahrhunderts unter dem Zeicchen des hellen Tages. Sie empfand aber die Helligkeit wie eine Ungunst des Schicksals. Auf dem Prinzip der Trennung, der Entasagung beruht das Wesen des menschen und die wirklichkeit des Ichs besteht bei ihnen aus dem Leben in der Angst. So finden wir in ihren lyrischen Gedicdten die Verlorenheit der Heimat des Glasubens, Gebrochenheit des harmonischen Lebens, wenn auch sie sich immer wieder an ihre Geistigkeit Klammern wollten.

심리훈련프로그램이 고혈압자의 안정시 심박수 및 혈압에 미치는 영향

최승욱,박철용,표내숙 한국스포츠심리학회 2003 한국스포츠심리학회지 Vol.14 No.2

본 연구는 복식호흡 및 점진적 이완훈련을 병행한 심리훈련으로 고혈압자의 안정시 심박수 및 혈압하감에 미치는 영향을 비교?검정하므로서 심리훈련이 고혈압자의 혈압하강에 효과적인 방안임을 제시하고자 실시된 실험 연구로서 중년 여성을 심리훈련군, 운동군, 통제군에 임의할당하여 4주간의 프로그램에서 실험실시 전, 실험중, 실험실시 후의 안정시 심박수 및 혈압을 측정하여 집단간 및 측정시기간의 변화를 비교하였다. 2002년 8월 5일부터 2002년 9월 6일까지 부산시에 거주하는 고혈압자로 40세에서 60세의 중년여성 21명을 대상으로 1주 동안 2회의 혈압 및 안정시 심박수를 측정한 후 심리훈련군은 주 2회 복식호흡 및 점진적 이완훈련을, 운동군은 유산소 운동을 주 3회 4주동안 실시하였고, 통제군은 어떠한 처치도 하지 않았다. 실험 2주 후 혈압 및 안정시 심박수를 측정하고, 실험 종료 후에 사후 조사를 실시하였다. 자료처리는 SPSS Program 10.0을 이용하여 집단간의 비교와 관련변수의 변화는 유의수준은 a=.05에서 다중 반복측정 분산분석(repeated measure ANOVA)을 이용하였고, 집단간 측정시기별 분석은 일원배치 분산분석(one-way ANOVA)을 이용하였으며 사후검증은 Tukey 방법(Tukey-test)에 의한 다중비교를 실시한 결과 다음과 같은 결론을 얻었다. 첫째, 안정시 심박수는 심리훈련군이 운동군과 통제군 보다 유의하게 감소하였다. 둘째, 수축기 혈압은 심리훈련군이 운동군과 통제군 보다 유의하게 감소하였다. 셋째, 이완기 혈압은 심리훈련군이 운동군과 통제군에 비해 점차적으로 감소하였으나, 통계적인 유의한 감소는 없었다. This research aims to prove that mental practice that includes abdominal breathing and progressive relaxation is an effective method in lowering the blood pressure by comparingㆍapproving the effect on the heart rate and the blood pressure under rest, which was proceed to divide middle-aged women into mental practiced group, aerobic trained group and controlled group randomly and to compare each group randomly and to compare each group and the period of measurement with the data of heart rate and blood pressure were measured before the test, during the test and after the test under rest from the program was performed for 4 weeks. The research was carried out from Aug. 05 to Sep. 06, 2002 with the measurement that includes the woman hypertensives from forties to sixties who reside in Busan and then, With the instruction of instruction of aerobic exercise 3 times a week for 4 weeks to aerobic trained group and non-instruction to controlled group. 2 weeks after the test, the blood pressure and heart rate were measured and post investigation was made after then. For the process of data using SPSS Program 10.0, repeated measure ANOVA was used at significant level ?=.05 in the comparison of each group and the change of relevant variable, one-way ANOVA was used in the analysis of the period of the test among each group and multiple comparison by Tukey test was made in the post approval. From this process of date, the following conclusion was drawn. First, the heart rate of the mental practiced group was decreased significantly in comparison to that of aerobic trained group and controlled group. Second, the systolic blood pressure of mental practiced group was decreased significantly in comparison to that of aerobic trained group and controlled group. Third, the diastolic blood pressure of mental practiced group was decreased steadily in comparison to that of aerobic trained group and controlled group, but statistical significant decrease was not found Consequently, its been proved that the mental practice which consists of abdominal breathing and progressive relaxation didn't affect on the diastolic blood pressure but did affect on the control of heart rate and the systolic blood pressure under rest.

Chitosan 및 Chitosan Lactate를 이용한 Glipizide의 서방형 제제설계

김상희,박승현,김하형,이광표 중앙대학교 약학연구소 1996 약학 논총 Vol.10 No.-

Glipizide(GPZ) is a second generation non-halogenated sulphonylurea hypoglycemic agent in maturity onset diabetes mellitus which stimulates the secretion of endogenous insulin from pancreatic β-cells. GPZ is almost completely absorbed with peak serum concentration being attained 1 to 3 hours after an oral administration. However, administration of GPZ at a high dose can occasionally induce the severe hypoglycemia, especially in the patients with renal impairment as well as impaired liver function due to some of the administered dose being excreted in the urine as unchanged drug and first-pass hepatic metabolism. Therefore, in order to develop an oral sustained-release preparation which reduces the side effect, chitosan and chtosan lactate were employed as a matrix for sustained-release tablet of GPZ in this experiment. Chitosan is a natural, biocompatible and biodegradable polymer, showing extrmemly low toxicity and it has been used as a vehicle for sustained felease preparations of drugs. Chitosan and chitosan lactate tablets of GPZ were prepared by directed compression using Hydraulic Lab. Press. at 340kg/cm^2 for 30sec. The tablet formulation of GPZ with chitosan alone ded not show sustained release properties at any concentrations(30-80%) of chitosan, but the release of GPZ from chitosan tablet containing citric acid shiowed prolonged characteristics. Chitosan readily forms a gel at lower pH, but shows poor gel-forming ability at higher pH. The effect of contents of chitosan lactate(10, 30 and 50%) or GPZ (5,10, 30 and 40%) on GPZ release profiles was studied. As the contents of chitosan lactate or GPZ increased, the release of GPZ decreased. And chitosan lactate showed sustained release properties even at lower concentration. In general, on the basis of dissolution kinetics, the release of GPZ from chitosan and chitosan lactate matrix tablet were followed Higuchi's square root of time equation and Bambe's equation. Therefore in conclusion, it might be possible to design the sustained release formulation of GPZ using chitosan and chitosan lactate.

높은 입체선택성을 나타내는 고리카보네이트의 친핵성 치환반응과 광학활성인 β-Hydroxy Ester의 합성

강석구,박동철,노호식,윤승현,홍덕표 성균관대학교 기초과학연구소 1995 論文集 Vol.46 No.1

The nucleophilic ring opening of the cyclic carbonates of optically active threo-2,3-dihydroxy esters afforded α-substituted β-hydroxy esters with highly regio- and stereoselectivity under mild conditions. The α-substituted β-hydroxy esters were transformed to β-hydroxy esters

만성폐쇄성폐질환의 중증도와 폐고혈압 및 폐성심으로 진행과의 상관관계

김형호,박철진,이준,장광표,하성일,이재록,하재화,권세훈,권용운,윤성호,이승일 朝鮮大學校 附設 醫學硏究所 2007 The Medical Journal of Chosun University Vol.32 No.2

Background: From many previous studies, Pulmonary hypertension is the known independent predictive factor of the mortality in COPD. Also pulmonary hypertension is the major cardiovascular complication of COPD and is associated with the progression to cor pulmonale and poor prognosis. Author want to analyze the correlation between pulmonary hypertension and the severity of COPD which are classified by forced expiratory volume in one second, and the extent of progression to corpulmonale. Methods: Retrospectively we investigated the medical records of 118 patients with COPD who had pulmonary function test and echocardiogaphy more than one times during one-year follow-up (from June, 2005 to May, 2006) at respiratory division in chosun university hospital, and then 50 patients were enrolled in this study. We classified the severity from FEVl in pulmonary function test based on the GOLD guideline, also this was comparably analyzed with RVSP, RVIDd, Visual Grading from echocardiography. Results: 16 patients out of 50 were classified as moderate severity, 25 patients were severe, and 9 patients were very severe group. RVSP was higher in more severe groups than less severe groups but when it comes to pulmonary hypertension that RVSP is over 35 mmHg, there was no valid difference lies in those groups. As FEVl decreases RVSP and RVIDd increases, and observed relationship between RVSP which was classified as below 35 mm and above 35 mm groups and visual grading which was classified as normal and dilatated groups revealed valid correlation, Conclusion: As pulmonary function decreases, progression to pulmonary hypertension and cor pulmonale increases but there was no remarkable difference of prevalence in above moderate severity groups. 연구배경 폐고혈압은 이전의 많은 연구에서 만성폐쇄성폐질환에서 독립적인 사망의 예측인자로 알려져 있다. 또한 폐고혈압은 만성폐쇄성폐질환의 주요한 심혈관계 합병증이며, 폐성심으로의 진행 및 불량한 예후와도 관련되어 있다. 이에 저자는 폐고혈압과 1초간 노력성호기량으로 분류되는 만성폐쇄성폐질환의 중증도 및 폐성심으로의 진행 정도를 비교 분석하여 이들의 상관관계를 밝혀보고자 한다. 방법 2005년 6월부터 2006년 5월까지 1년 동안 본 병원 호흡기 내과에서 폐기능검사와 심장초음파 검사를 각각 1회 이상 시행한 적이 있는 118명에 한하여 의무기록을 후향적으로 조사하여 이 중에서 50명을 대상으로 하였다. 폐기능검사 시행 결과 얻은 1초간 노력성호기량을 바탕으로 GOLD guideline에 따라 중증도를 분류하였으며 이를 심초음파 시행 결과 얻은 우심실수축기압, 확장기말 우심실내부간격, 시각등급과 비교분석하였다. 결과 대상 환자 50명중 16명은 중등증, 25명은 중증, 9명은 최고중증 그룹으로 분류되었다. 우심실수축기압은 중증도가 높은 그룹에서 낮은 그룹에 비해 더 높게 나타났으나 폐고혈압에 해당하는 우심실수축기압이 35 mmHg이상인 환자에 있어사는 각 그룹간에 유의한 차이를 보이지 않았으며, 확장기말 우심실내부간격, 시각등급 또한 중증도와 유의한 차이를 보이지 안항ㅆ다. 1초간 노력성호기량이 감소함에 따라서는 우심실수축기압, 시각등급은 증가하였으며 우심실수축기압을 35mmHg 미만과 이상인 그룹으로, 시각등급을 정상과 확장이 있는 그룹으로 분류하여 비교한 결과에서는 유의한 상관관계를 나타냈다. 결론 폐기능이 감소할수록 페고혈압 및 폐성심으로의 진행이 증가하였으나 폐기능검사에서 중등도 이상의 그룹에서 유병률의 차이는 크지 않았다.

Dimethyl-β-Cyclodextrin을 이용한 Omeprazole의 대체투여경로 개발

정미분,박승현,김하형,이광표 중앙대학교 약학연구소 1996 약학 논총 Vol.10 No.-

For the purpose of increaing the stability and bioavailability of omeprazole(OME), inclusion complexes of OME with demethyl-β-cycoldextrin(DMβCD) and hydroxypropyl-β-cyclodextrin(HPβCD) were prepared by freeze drying method. The interactions of OME with DMβCD and HPβCD in aqueous solutions and in solid states were investigated by solubility study. IR spectroscopy and DSC. The type of OME-DMβCD inclusion complex was found to be classified as Bs-type on phase solubility diagram, and the stoichiometric ratio of OME-DMβCD incousion complex was a 1:2 and stability constant was 359.46M^-1. The OME-HPβCD inclusion complex appeare to be AL type. The stoichiometric ratio was found to be 1:1, and stability constant was 34M^-1. Clear differences in a IR spectra and DSC curves were observed between the inclusion complex formation at pH 6.8. The mean dissolution time(MDT). which represents the degree of the rapidity of dissolution rate, was increased in the order of OME-DMβCD I.C(0.60) < OME-HPβCD I.C.(0.71) < OME-DMβCD P.M.(7.23) < OME-HPβCD P.M.(10.06). In order to compare simultaneously the intrinsic absorptivity of OME in the jejunum and nasal cavity of rats, the in situ nasal and jejunal perfusion method was employed as a new trial. The in situ recirculation study revealed that OME-CDs inclusion complexes rate proportionally increased the absorption of OME in nasal cavity(N) and the jejunum(J) of rats. The decreasing order of apparent OME permeability(Papp:cm/sec×10-5), corrected for surface area of absorption was as follows : 8.77±0.18(N, OME-DMβCD). > 5.97±0.17(N. OME-HPβCD) > 4.84±0.02(J. OME-DMβCD). 3.50±0.25(J. OME-HPβCD) at IPB pH 7.4. The present results suggest that OME-DMβCD inclusion complex may serve as a potential means for the nasal delivery of OME.

한우 체외수정란의 동결보존 및 핵이식에 관한 연구 : Ⅱ. 체외수저란의 동결방법이 동결.융해후 생존율에 미치는 영향 Ⅱ. Effects of Survival Rates of Frozen-Thawed Embryo Frozen Methods

정장용,박희성,홍승표,김정혁,桑潤滋,離素娟 진주산업대학교 농업기술연구소 1999 農業技術硏究所報 Vol.12 No.-

본 연구는 수정란의 동결기법을 확립하고자 배반포기에 있는 한우 체외수정란을 이용하여 유리화 및 직접이식 동결방법으로 동결을 실시하여 동결·융해후 생존성, 동해방지제의 종류, 평형 및 할구의 손상여부 등에 대한 영향을 조사한 결과는 다음과 같다. 유리화 동결방법중 GESD-VS와 EFS 용액을 동해방지제로 사용하였을 때 동결·융해 수정란의 생존율은 각각 69.8 및 37.3%로써 이들간에 유의적(P<0.05)인 차이가 있었으며, 동결기를 이용한 직접이식 동결법은 생존율이 56.0%로써 유리화 동결방법과는 유의적(P<0.05)인 차이가 없었다. 동결·융해후 생존한 수정란의 발달율은 GESD-VS용액을 사용하였을때가 53.4%로써 EFS용액을 사용하였을때의 24.0%보다는 유의적(P<0.05)으로 높게 나타났다. 1 및 2단계 평형시 단계별 생존율은 각각 11.7 및 23%로써 이들간에 유의적(P<0.05)인 차이가 없었으며, 동결·융해 수정란의 발달율도 각각 10 및 16.8%로써 평형방법간에 유의적(P<0.05)인 차이가 없었다. 신선수정란, 유리화 동결수정란 및 동결비를 이용한 직접이식 동결법으로 동결하였을 때 동결·융해 수정란의 할구수는 각각 98.7±2.3개, 69.1±2.6 및 84.2±2.7개로써 동결방법들간에 유의적(P<0.05)으로 할구수의 차이가 많았다. This study was carried out to investigate the effect of cryoprotectants, equilibration step, number of blastomeres on survival and developmental rates of bovine in vitro fertilized blastocysts(Korean native cattle) frozen/thawed by vitrification and frozen for direct transfer(1.8M ethylene glycol). The bovine oocytes were collected from 2-5mm follicles, matured for 24hrs in 5% CO_2 incubator and then fertilized with cauda epididymis sperm. Blastocyst at day 7-9 ater IVF were frozen in vitrification and frozen for direct transfer(1.8M ethylene glycol). Survival and developmental rates of frozen-thawed IVF embryos were examined. The results were as follow ; The survival rate of IVF blastocyst after vitrification in GESD-VS, EFS and 1.8M ethylene glycol solution were 69.8, 37.3 and 56.0%, respectively(P<0.05). When equilibrated in 1 step(11.7%) was similar to the result from 2 step(23.0%), and no significantly(P<0.05) different between the equilibrated steps. The number of blastomere at blastomere at blastocyst of bovine embryos frozen/thawed in fresh, vitrification and freezing for direct transfer were 98.7±2.3, 69.1±2.6 and 84.2±2.7, respectively, and were significantly(P<0.05) different between the freezing methods.