Spermatogonial stem cells (SSC) undergo self-renewal division and generate spermatogenesis to produce mature spermatozoa. Very recently in some species, include rodent and farm animals, SSC has been isolated and cultured in vitro. In this study, we analysed SSC marker of both 5 days old and pubertal pig testis by histological method. In 5day pig testis, prior to set of spermatogenic differentiation, the seminiferous tubules contain a relatively large number of SSCs than in pubertal pig testis. Then putative pig SSCs were successfully isolated from 5 day pig testis, and cultured long term using CD34 positive cell culture media contained GDNF, bFGF, LIF and EGF. The SSC colonies were appeared at 3 days after cells were seed. The SSC colonies were alkaline phosphatase positive and strongly expressed PGP 9.5, PLZF and DBA, but not expressed GATA4 and LH receptors. The SSC colonies were stably proliferated in GDNF, bFGF, LIF and EGF contained CD34 positive cell culture media up to 60 days. This study will be facilitated to study of in vitro and ex vivo spermatogenesis and of production of transgenic pigs using sperm vector.

INTRODUCTION In rodent, molecular markers of spermatogonia, spermatocyte, spermatid and sperm have been identified. It has been reported that DBA, PGP 9.5 and NanpG can be the markers for spermatogonia in pig. For further understanding the spermatogenesis of the pig on morphological and molecular level, we report identification of testicular cells in neonatal and pubertal pig testis, and a putative marker for spermatogonia and spermatid in pig testis. MATERIALS AND METHODS Neonatal (3 day) and pubertal testis (150 day) was cut and fixed in Bouin’s solution for immunohistochemistry (IHC). Gonocytes were isolated from neonatal testis for immunocytochemistry (ICC). Based on references (Phillips et al., 2010), thirteen antibodies (VASA, Oct4, NanoG, PGP 9.5, DAZL, SCF, GFR-alpha 1, PLZF, c-kit, integrin-beta1, Thy1, Sohlh1 and CD9) were used for IHC and ICC. Paraffin section was performed for IHC. Gonocytes were attached to the APS-coated slides for ICC. HRP-conjugated and florescent-labeled secondary antibody was used for IHC and ICC, respectively. RESULTS In histological analysis, spermatogonia and Sertoli cells, which are enclosed by seminiferous tubule and connective tissue, were observed in neonatal testis. However, complete spermiogenesis, including spermatocyte, spermatid and spermatozoa, was not observed in neonatal testis. Spermatocyte, spermatid and elongated spermatid were observed in pubertal testis but matured spermatozoa were not observed. As a result, two antibodies (PGP 9.5 and GFR-alpha1) of thirteen antibodies were available for IHC and ICC. As reported in other studies, PGP 9.5 protein was detected in spermatogonia of ne-onatal in IHC. In addition, it was observed in spermatogonia of pubertal testis. GFR- alpha1 protein was detected in spermatogonia of neonatal testis and spermatid of pubertal testis. In ICC, PGP 9.5 protein was detected in gonocytes as reported in other studies. GFR-alpha1 was also observed in gonocyte isolated from pig testis. In this study, we found that 1) only spermatogonia exist in neonatal pig testis (3 day), 2) GFR-alpha1 is a new marker for spermatogenesis in pig testis.

Interferon induced transmembrane protein-1 (IFITM1) is one of transmembrane protein which is differentially expressed in uterus during estrus cycle and pregnancy, that IFITM1 gene is highly expressed in estrus stage by the effect of estrogen, and in parturition by the effect of PGF2 alpha. This genes are also up-regulated in cells with hyperactivation of the WNT/β-catenin signaling pathway. In this study, to identify a function of IFITM1, the binding partner of IFITM1 were determined using immunoprecipitation and LC- MASSMASS methods. 1, 3 and 5 ug of polyclonal anti-IFITM1 antisera were used for immunoprecipitation, and the 75 kDa of specific band was detected in silver stained polyacylamide gel. This band were chracterized using LC-MASS-MASS, and revealed this band is glucose regulated protein 75 (GRP75) which binds to p53 and inhibits the p53 action in nucleus. To identify the localization of GRP75 in cells, immunocytochemical approach has been applied, and GRP75 is expressed in mitochondria of L929 murine connective tissue cells. Co-localization study between IFITM1 and GRP75 in L929 cell identified that these two proteins were closely expressed in mitochondria. Although the role of the interaction of these two protein need to be clarified in various biological phenomena, this data suggest that close interaction of IFITM1 and GRP75 may regulate cellular functions in uterus on sets of estrus cycle and pregnancy.

SERPINB3 (also known Squamous cell carcinoma antigen 1, SCCA1) is involved in apoptosis, immune response, cell migration and invasiveness of cells. It has been investigated in various types of squamous cell carcinoma. Therefore we investigated the functional role of SERPINB3 gene in human epithelial ovarian cancer (EOC) using laying hens, the most relevant animal model. In 136 laying hens, EOC was found in 10 (7.4%). We compared the expression and localization of SERPINB3 using RT-PCR, quantitative RT-PCR, in situ hybridization and immunohistochemistry, and SERPINB3 activation was detected in chicken and human ovarian cancer cell lines using immunofluorescence microscopy. Thereafter, we examined the prognostic value of SERPINB3 expression in patients with EOC by multivariate linear logistic regression and Cox’ proportional hazard analyses. In present study, SERPINB3 mRNA was induced in cancerous ovaries (p< 0.01), and it was only expressed in the glandular epithelium(GE) of cancerous ovaries of laying hens. SERPINB3 protein was localized predominantly to the nucleus of glandular epithelium in cancerous ovaries of laying hens, and it was abundant in the nucleus of both chicken and human ovarian cancer cell lines. In 109 human patients with EOC, 15 (13.8%), 66 (60.6%) and 28 (25.7%) of those patients showed weak, moderate and strong expression of SERPINB3 protein, respectively. Strong expression of SERPINB3 protein was a prognostic factor for platinum resistance (adjusted OR, 5.94; 95% Confidence Limits, 1.21-29.15). Therefore SERPINB3 may play an important role in ovarian carcinogenesis and be a novel biomarker for predicting platinum resistance and a poor prognosis for survival in patients with EOC. This research was funded by the World Class University (WCU) program (R31-10056), Basic Science Research Program (2010- 0013078) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology and by the Next-Generation BioGreen 21 Program (No.PJ008142), Rural Development Administration, Republic of Korea.

Rabies is one of the most dreadful diseases known to human. Annually, more than 55,000 human deaths occur throughout the world. The main transmitters are dogs. In South Korea, urban rabies is eliminated after massive national vaccine programme but rabies is still present in wildlife around northern part of the country near the border. Occasionally, rabies cases are still reported and there are spill over cases from racoon dogs. No human case was reported since 2005. Therefore, risk of rabies from exporting domestic dogs and cats from South Korea is very low. Hence, foreign rabies can be introduced by importing wild carnivores and unvaccinated dogs and cats under the age of three months since the South Korean legislation does not cover them. Therefore, it is essential to update current import regulation to minimise the risk of rabies.

A new cultivar "KNR11-1" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2598 and KNR2610. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15 - 16℃. The period of harvesting including primordia formation of "KNR11-1" was one day faster than that of control strain KNR2312. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has the teeth of a comb. The yield was 90.4 g/850cc plastic bottle. The fruit body has a longer shelf life than control strain KNR2312 at 4℃. It is able to store up to 67.5 days in a cold room. Analysis of the genetic characteristics of the new commercial strain "KNR11-1" showed a different profile as that of the control strain KNR2312 when Random Amplified Polymorphic DNA primers were used. The new cultivar of Pleurotus eryngii is characterized by the improved storability after harvesting.

Serpins are a superfamily of related protease inhibitors with common structural features and inhibitory mechanisms. However, SERPINA 14 in mammals does not have inhibitory activity against most known proteases. Rather, it may have an immunoregulatory role in mammals to prevent rejection of the fetal allograft by inhibiting lymphocyte proliferation and natural killer cell function. In the pig, SERPINA14 is involved in iron transport to the fetus by binding to and stabilizing the iron-binding protein uteroferrin (ACP5). In chickens, these very little known about serpins in chickens. Therefore, we investigated the expression patterns of serpin genes in the oviduct of adult hens and in the oviduct of 37-day-old chicks treated with an estrogen analogue, diethylstilbestrol (DES). Results indicated that SERPINB3 and SERPINB11 genes were highly expressed in oviducts of DES-treated chicks, but not in oviducts of control chicks. Both SERPINB3 and SERPINB11 transcripts were localized specifically to the gland-like areas of oviducts of DES-treated chicks. Immunohistochemical analyses confirmed that SERPINB3 and SERPINB11 proteins were present in the gland-like area and luminal epithelium of the oviducts of DES-treated chicks. Collectively, the results suggest that SERPINB3 and SERPINB11 are expressed in response to estrogens and they have distinct functions related to development and differentiation of the mature oviduct in hens.

S-adenosylhomocysteine hydrolase-like protein 1 (AHCYL1), also known as IP3 receptor- binding protein released with IP3 (IRBIT), is a member of the AHCY-like protein family. AHCYL1 protein regulates IP3-induced Ca2+ release in the cytoplasm of cells and, therefore, is likely to be an important gene regulating various biological processes in the oviduct of chickens. Inmammals, expression is greatest during activation of dendritic cells which are antigen presenting cells associated with immunoregulatory processes in blood and skin. However, the identification of the AHCYL1 gene in chickens has not been investigated. In the present study, we first used RT-PCR to demonstrate AHCYL1 gene expression in adult chicken organs and oviducts of immature chickens treated with DES (diethylstilbesterol, a synthetic estrogen agonist). The results indicated that AHCYL1 mRNA is expressed in chicken reproductive organs (testis, ovary and oviduct). Inaddition, expression of AHCYL1 mRNA increased in response to DES-treated immature oviducts compared to the non-treated control immature oviducts of chickens. Interestingly, AHCYL1 was abundant in the cytoplasm of luminal and glandular epithelia, but not in other cell-types such as stroma and connective tissues, of the chicken oviduct. These results suggest that AHCYL1 is a novel estrogen-stimulated gene associated with development of the chicken oviduct, as well as functions of oviductal glandular and luminal epithelia that may include activation of resident immune cells, such as dendritic cells.

Mitochondria diseases have been reported to involve structural and functional defects of complex I-V. Especially, many of these diseases are known to be related to dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I). The dysfunction of mitochondria complex I is associated with neurodegenerative disorders, such as Parkinson's disease, Huntington's disease, and Leber’s hereditary optic neuropathy (LHON). Mammalian mitochondrial proton-translocating NADH–quinone oxidoreductase (complex I) is largest and consists of at least 46 different subunits. In contrast, the NDI1 gene of is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. The gene using a recombinant adeno-associated virus vector (rAAV-NDI1) was successfully expressed in AML12 mouse liver hepatocytes. The NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the gene failed to survive. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced cells was not affected by rotenone which is inhibitor of complex I, but was inhibited by antimycin A. Furthermore, these results shown that Ndi1 can be functionally expressed in the AML12 mouse liver hepatocytes. It is conceivable that the gene is powerful tool for gene therapy of mitochondrial diseases caused by complex I deficiency. In the future, I will attempt to functionally express the NDI1 gene in mouse embryonic stem (mES) cell.

A member of mice interferon inducible transmembrane protein like family, IFITM1, is reported to play an important role in primordial germ cell (PGC) formation and this protein reported for anti‐proliferation. This study conducted to investigate the mIFITM1 expression in reproductive organs of male mice. mIFITM1 expression in testis and epididymis were revealed by immunostaining and immuno blot. Moreover, we showed that mIFITM1 is related to development of mouse testis. mIFITM1 protein expression as markedly upregulated around 5‐ 15day after birth in testis, but postnatal 21 56 day detected pattern was decreasing by immunoblot. In the other reproductive organ, epididymis, we observed that mIFITM1 protein was strongly expressed in caput, corpus and cauda. We analyzed IFITM1 gene expression under conditions of androgen manipulation. Total RNAs were obtained from the epididymis of adult mice that castrated and injected. Testosterone replacement for animals 7day after surgery. In castrated animals, A clear decrease was found in the IFITM1 protein level on Interestingly, castrated mice was revealed that mIFITM1 expression is strong. At that time, the injected catration mice decrease in IFITM1 gene expression. We also found same result from the immunoblot analysis. Our data suggest that the function of the IFITM1 expression in sperm is remain unclear but mIFITM1 expression will be important to postnatal development of mice testis and spermatogenesis. Also, mIFITM1 regulated not only sexual maturation by gene expression in the reproductive organs but also by hormone.

The occurrence and rapid range expansion of a fulgorid, Lycorma delicatula(White), has recently been reported in korea. It was previously known to occur in China, Japan, Vietnam and India. The first occurrence fo L. delicatula was reported in 2004 in the Cheonan. It has one generation per year and overwinters as egg masses on the bark of host trees. The number of egg per egg mass was 32.69±6.49. The cumulative ratio of hatchability at different temperature was the highest at 25℃ and the lowest at 10, 30℃. The egg hatchability was investigated in different regions in Chung-buk. Since L. delicatula eggs has been stored in -20±1, 0±1, 5±1℃ conditions for 1 ∼60 days, the cumulative ratio of hatchability was 2% stored in -20±1℃ for 3days, But hatchability was 52, 48% stored in 0±1℃ for 3, 14days and 82, 68% stored in 5±1℃ for 3, 14days. And hatchability was 0% for 60days. By straight regression equation(the growth ratio/treatment degree) the growth zero degree of L. delicatula was 10.4℃. The hatchability of L. delicatula in Cheongwon-gun, Okcheon-gun, Cheongju-city where’s winter lowest temperature was over -19℃ was over 79%. But The hatchability of L. delicatula in Jincheon-gun where’s winter lowest temperature was less than -19℃(continuos two days) was 35%.

The notion that dental amalgam is a potential source of mercury exposure remains a controversial issue. However, there are few epidemiological analyses that have addressed whether this occurs in children. We aimed in our current study to identify the relationship between dental amalgam filling surfaces and the blood mercury levels in a cohort of 711 South Korean children aged between 8-9 years. Oral examinations were conducted to detect the number of amalgam filling surfaces on the teeth of these individuals. Blood samples were also taken from these children to assess the levels of mercury accumulation in the body. The amalgam filling surfaces were classified into four groups based on their number: 0 (n = 368), 1-5 (n = 219), 6-10 (n = 89), and 11+ (n = 35). The blood mercury levels in the children with more than 10 amalgam surfaces was 0.47 μg/L higher on average than those with no amalgam surfaces after adjusting for the frequency of fish or seafood consumption, age, and gender (P < 0.05). We found from our data that a higher number of dental amalgam fillings correlated with a higher blood mercury level in Korean children. Further studies are needed to investigate whether these elevated mercury levels exert neurotoxic or nephrotoxic effects.

A new cultivar "KNR10-1" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2594 and KNR2610, respectively. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15 - 16℃. The period of harvesting including primordia formation of "KNR10-1" was one day faster than that of control strain KNR2312. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has the teeth of a comb. The yield was 90±12.9g/850cc plastic bottle. The fruit body has a longer shelf life than control strain KNR2312 at 4℃. Analysis of the genetic characteristics of the new commercial strain "KNR10-1" showed a different profile as that of the control strain KNR2312 when RAPD (Random Amplified Polymorphic DNA) primers were used. This new variety of King Oyster mushroom is characterized by the improved storability after harvesting.

In East-Asia, before modern times, writings in several countries were written in Han Character. At that time, because the world of East-Asia belonged to Han Character and Classics cultural sphere, for the political, diplomatic and cultural communication naturally they needed Han Character writing. This Han Character writing were used around nobility. The middle ages was a society for nobility, thus language of ruling activity was mainly Han Character. As a result, the lower class was neglected from Han Character writing, the language of ruling class. Until 20th century the countries in East-Asia that wrote sentences in Han Character were China, Korea, Japan, Vietnam and so on. Of course, although these countries used Han Character to write, each level, contents and historic process of Han Classics are different. But there was no difference in that they had used sentences in Han Character. From the beginning of 20 century, all of east-Asia countries writing pattern and education system going different own way. However, now in the each East-Asia country, Han character and subject (old classics with Han character) education is very important. Therefore, we need to do mutual recognition about language education in Asian sphere have Han Character in common being developed newly .Also, we need to recognize that not only Han Character vocabulary but also Han Classics have a connection and exchange studies about it. I hope in this symposium, we need to the chance to understand new relation sharing 'Han Character vocabulary' in Asian sphere. We can expect scientific development through trades of various learning and education methods or theory about Han Classics and Han Character in each country. So, I suggest as follows for this exchange. 1. Holding international conference on learning and teaching of Han Characters in Asian languages regularly. Annually or biennially, holding a symposium which have principles of own expense, attending 10 or so representing each countries. 2. Carrying out common study to develop. It is hard for the moment , building a database of basic data for each countries' formal change and sentence study and developing a database of Han Classics and Han Character 3. Concluding simple treaty about international exchange among the scholars who attended to conferences. That is, it is not only symposium but exchange between department and department or department and faculty. 4. Publishing international academic journal in AHCI level which represent Han Classics and Han Character education in Asian Han Character culture sphere. In this case, it is not around certain society in certain country but is around scholars and society in Asian sphere. Fifth, holding a conference in circulation system to open and manage the homepage concerned about this. In homepage, it will offer the contents about symposium, notice and results, collecting project, VOD service about key note lecture during symposium, links of each country's society and thesis services published at international academic journal and so on. These are can be hard for the moment, if we push ahead actively, we can do them. Just, it is important we share what recognition for ourselves. Without forming a bond of sympathy, international can be stopped at one-off event. Therefore I suggest exchange with the suggestions . Of course we can't accomplish all of them at once. It can be desirable that we expects gradual development by starting from the part of them. Finally, study and exchange through this symposium doesn't stop at self-contentment but is for the development of Asia for future and peaceful bonds.