在近几年的留学生文言文教学中，我们发现了留学生学习文言文的一些困难，对此我们做了很多调研和总结。本文认为在留学生的文言文教学中，首先应遵循4个一般原则：第一，解决文字问题，识字是读文言文的基础，要让学生能够识别古字、讹字、通假字等；第二，掌握一定的词汇，了解古今词义的差别，知道词义具有社会性、时代性、系统性；第三，了解汉语语法的特点，知道汉语句法、词法的特殊性，并且古今汉语语法的差异如判断句的表述等；第四，有一定的古文化常识和丰富的历史知识，了解中国朝代变迁和文化习俗，如避讳等等。另外，由于教育对象的变化，在留学生文言文教学中，我们还要遵循其特殊原则，那就是加强古文化常识和古史的介绍。期望这些来自教学实践的研究所得会对留学生文言文教育有所帮助。

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii- specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was as- sessed against 43 strains of mitis group streptococci, in- cluding clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amp- licons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17- F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

An entomopathogenic bacterium, Xenorhabdus nematophila (Xn), is symbiotic to a nematode, Steinernema carpocapsae, and exhibits high pathogenicity to lepidoptera insects. Its metabolites released into the bacterial culture broth and also virulent in oral especially when they are treated with Bacillus thuringiensis (Bt). This study devised a high efficacy microbial insecticide by combining Xn culture broth and Bt. Bt kurstaki (Btk) exhibited relatively higher pathogenicity to Plutella xylostella than Spodoptera exigua larvae. By contrast, Bt aizawai (Bta) showed a reverse pathogenicity pattern. Phase Ⅰ type of Xn (XnK1) was isolated from S. carpocapsae Pochun and exhibited high pathogenicity than phase Ⅱ bacteria. Three bacterial mixtures of Bta+XnK1, Btk+XnK1, and Bta+Btk+XnK1 were prepared and analyzed in their target insects. Bta+XnK1 showed higher pathogenicity than those of Bta alone or Btk+XnK1 in P. xylostella. Btk+XnK1 showed higher pathogenicity than those of Btk alone or Bta+XnK1 in S. exigua. Bta+BtK+XnK1 showed high pathogenicity against both P. xylostella and S. exigua.

Although assisted reproductive technology is very useful to develop novel and therapeutic biomaterials for reproduction, research on molecular mechanism of folliculogenesis in pig is not clear. Therefore, the alteration of gene expression during follicular development in pigs was examined in this study. The expression of folliculogenesis-related genes was quantified in preantral (250～300 μm) and antral (>300 μm in diameter) follicles, and overall gene expression was evaluated by a genome-wide microarray. The microarray results showed that 219 genes were differentially expressed, and of those, 10 and 22 known genes showed higher and less expression at the preantral stage than at antral stages, respectively. Among them, the expression of NR0B1, PPARG, GATA4, and ANXA2 genes related to folliculogenesis was validated by quantitative real-time PCR analysis. The expression of PPARG and GATA4 genes were increased at antral stages, but a significantly stage-specific increase (p<0.05) was only detected in annexin A2 (ANXA2) in antral-stage follicles. The expression of NR0B1 genes was increased at preantral stage and these patterns of gene expression were comparable to the results obtained by microarray analysis. We propose that the systematical regulation of genes supporting specific follicle stage should be employed for improved in-vitro folliculognesis.

본 조사에서 확인된 관속식물상은 98과 310속 402종 52 변종 7품종 1아종 462종류로 확인되었다. 한반도 고유종은 국립생물자원관(2011)이 지정한 한반도고유종은 눈주목, 외대으아리 및 할미밀망 등 총 10종류가 확인되었다. 식물 구계학적 식물종은 Ⅴ등급 종은 목련과 왕벚나무로 2종, Ⅳ등급종은 산복사, 측백나무 및 꽃창포 등 3종, Ⅲ등급종 은 단풍나무, 탱자나무 및 노각나무 등 3종, Ⅱ등급 종은 석창포, 주목 및 개고사리 등 3종 그리고 Ⅰ등급 종은 이팝 나무, 회양목 및 거지덩굴 등 8종이 확인되었다. 전주지역 소류지 7곳에서 부유식물은 개구리밥과 좀개구리밥으로 2 분류군, 부엽식물은 마름, 애기마름 및 가는가래 등 6분류 군, 침수식물은 말즘과 물수세미로 2분류군 그리고 정수식 물은 고마리, 왕버들 및 논냉이 등 총 52분류군이 확인되었 다. 귀화식물종은 총 45종류로 국화과가 노랑코스모스, 미 국가막사리, 실망초 및 개망초 등 16종으로 가장 많았고, 다음은 벼과로 메귀리, 큰김의털 및 들묵새 등 7종이, 메꽃 과 4종, 현삼과 3종, 마디풀과, 명아주과, 십자화과, 콩과 및 바늘꽃과는 각 2종 그리고 자리공과, 석죽과, 비름과, 대극과 및 소태나무과 등은 각 1종씩 확인되었다. 환경부 생태계교란식물은 돼지풀, 털물참새피와 물참새피 3종류가 출현하였다. 수질은 COD 기준으로 10이 초과되는 소류지 는 구주제가 16.23, 성곡제 14.59, 황학제 13.79 및 오송제 10.66으로 매우 나쁜 상태를 보였으며, 10이하인 소류지는 지사제 9.48과 서은제 8.65로 나쁜 상태이며, 학소제만 4.74 로 보통상태였다. 소류지 생태공원 이용실태 응답자의 50% 이상이 주 2회 이하로 방문하는 것으로 응답하였으며 생태 공원을 이용함에 있어 가장 큰 이점이 약 50%의 응답자가 스트레스 해소 및 생활 활력의 증가를 꼽았다. 탐방객 중 한 부류는 위락, 편의시설이 잘 구비된 곳보다는 불편하더 라도 한적한 곳을 찾고 싶다는 응답자가 50%를 넘었다.

The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

Germ cell-specific hyaluronidases such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5) are in part responsible for dispersal of the cumulus cell mass, which is a critical step in establishing fertilization in mammals. In this study, we identified two testis-hyaluronidases, SPAM1 and Hyal5, in hamster and rat. These two genes were expressed specifically in the testis. At the protein level, hamster SPAM1 and Hyal5 display 78.7% and 75.4% identity with mouse SPAM1 and Hyal5. Further, the activity of the enzymes with respect to cumulus cell dispersion did not differ, although we observed that the enzymatic activity differed in pH range. These studies suggest that different sperm hyaluronidases are capable of dispersing the cumulus cell mass despite differences in enzyme activity.

느타리 버섯류의 새로운 품종을 개발하기 위하여 고품질의 짙은 회색느타리 신품종을 육성하였다. 2003년부터 2004년까지 느타리 유전자원의 특성검정을 하였다. 2008년에 육종모본수한과 ‘농기201호’의 단핵체간 교잡하여 04-154 교잡주를 육성하였다. 이 교잡주 04-154와 품종 ‘청풍’의 단핵체와 다시 교잡하여 우수한 Po2008-275를 선발하여 특성검정, 확대재배를 실시하여 농작물 직무육성 신품종 선정심의회에서 ‘구슬’로 명명되었다. 주요특성으로 균사 생장 적온이 25~30℃이며 버섯 원기형성 및 발생 온도는 10~16℃이였다. 자실체의 갓 색깔은 짙은 회색으로 자연 상태에서 봄, 가을에 재배가 알맞은 특성을 가지고 있다. 균사체 배양기간은 25~30일이며 균 긁기 후 초발이소요일수는 3~5일로 온도가 높을수록 단축된다. 자실체형태는 얕은 깔때기형이다. 대굵기는 16.8±1.6㎜, 대길이는 51.0±4.4㎜로 다른 느타리 종에 비해 자실체 대가 굵고 다소 짧은 편이다. 자실체 수량은 상자 당(43 x 43 x 11cm) 1,545g으로 04-154 계통친이 100일 때 ‘구슬’ 137, ‘청풍‘이 146이었다. 가변특성으로는 감자배지와 버섯완전배지에서 균사를 배양한 결과 버섯완전배지에서 생장이 양호하였다. 또한 3종류의 primer를 이용하여 새로운 품종 ‘구슬’과 모균주에 대한 DNA profile을 분석한 결과 primer URP 1, primer URP 2, primer URP 5에서 양친주의 주요 밴드를 가지며 대조구인 ‘춘추2호’와는 뚜렷하게 구분되었다. 신품종 느타리 ‘구슬’은 소비자들이 가장 선호하는 짙은 회색의 갓을 나타내어 고품질을 요구하는 소비자를 만족시키는데 기여 할 것으로 기대된다.

Egg yolk immunoglobulin (IgY) is the antibody in egg yolk and can be produced in egg yolk by immunizing hens with antigens. IgY is functionally equivalent to mammalian IgG. It is found in the serum of the chicken and is passed from the mother chicken to the embryo via the egg yolk, a process that results in a high concentration of IgY in the egg yolk. The objective of this study was to evaluate the efficacy of specific IgY against enterotoxigenic Escherichia coli (ETEC) K99, Salmonella Typhimurium and Salmonella Choleraesuis that cause porcine bacterial diseases. To prepare specific IgY, Hy-Line Brown chickens were vaccinated with killed vaccine complex including E. coli K99, S. Typhimurium and S. Choleraesuis. The chicken egg yolk antibodies were purified from egg yolk by ammonium sulfate precipitation and the quality of the final preparation was confirmed by sodium dodecyl sulfate polyacrylamide gel electro-phoresis (SDS-PAGE). Titres of specific IgY in final preparations were measured by an enzyme-linked immunosorbent assay (ELISA). Antibody titers peaked at 3 weeks regardless the bacterial types and the similar patterns of immune response were observed for respective pathogens. In growth inhibitor test, specific IgY showed inhibitory effect on bacterial growth. After 0, 3, 6 and 12 hour of incubation with specific IgY (100 ㎍/㎖, 250 ㎍/㎖, 500 ㎍/㎖), there was a significant decrease in the growth (A 600nm ) of E. coli K99, S. Typhimurium and S. Choleraesuis compared to nonspecific IgY and controls. In BALB/c mice, the effect of specific IgY (100 ㎎/㎏, 250 ㎎/㎏) on bacterial challenges was investigated by intramuscular injection and oral administration of bacteria. Mice treated with specific IgY showed high survival rate though there was no significant differences on blood biochemichal examinations between treated and untreated groups. These results indicate the potential of specific IgY for the treatment of porcine bacterial diseases caused by E. coli K99, S. Typhimurium and S. Choleraesuis.

In this study, a method was developed using molecular biological technique to distinguish an authenticity of meats for processed meat products. The genes for distinction of species about meats targeted at 12S or 16S genes in mitochondrial DNA and the species-specific primers were designed by that PCR products' size was around 200bp for applying to processed products. The target materials were 10 species of livestock products and it checked whether expected PCR products were created or not by electrophoresis after PCR using species-specific primers. The results of PCR for beef, pork, goat meat, mutton, venison, and horse meat were 131, 138, 168, 144, 191, and 142 bp each. The expected PCR products were confirmed at 281, 186, 174, and 238 bp for chicken, duck, turkeymeat, and ostrich. Also, non-specific PCR products were not detected in similar species by species-specific primers. The method using primers developed in this study confirm to be applicable for composite seasoning including beefs and processed meat products including pork and chicken. Therefore, this method may apply to distinguish an authenticity of meats for various processed products.

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.

Peroxiredoxin II (Prdx II; a typical 2‐Cys Prdx) has been originally isolated from erythrocytes, and its structure and peroxidase activity have been adequately studied. Mice absent to Prdx II proteins had heinz bodies in their peripheral blood, and morphologically abnormal cells were detected in the dense red blood cell (RBC) fractions, which contained markedly higher levels of reactive oxygen species (ROS). In this study, a labeling experiment with the thiol‐modifying reagent biotinylated iodoacetamide (BIAM) in Prdx I‒/‒ mice revealed that a variety of RBC proteins were highly oxidized. To identify oxidation‐sensitive proteins in Prdx II‒/‒ mice, we performed RBC comparative proteome analysis in membrane and cytosolic fractions by nano‐UPLC‐MSE shotgun proteomics. We found oxidation‐sensitive 54 proteins from 61 peptides containing cysteine oxidation, and analyzed comparative expression pattern in healthy RBCs of Prdx II+/+ mice, healthy RBCs of Prdx II‒/‒ mice, and abnormal RBCs of Prdx II‒/‒ mice. These proteins belonged to cellular functions related with RBC lifespan maintain, such as cytoskeleton, stress‐induced proteins, metabolic enzymes, signal transduction, and transporters. Furthermore, protein networks among identified oxidation sensitive proteins were analyzed to associate with various diseases. Consequently, we expected that RBC proteome may provide clues to understand redox‐imbalanced diseases.

Despite of the absence of hyperacute rejection and acute humoral xenograft rejection, the organ graft of the a1,3-galactosyltransferase (GalT) gene knockouted (KO) and complement regulatory protein (CRP) expressing pig into a nonhuman primate is rejected by development of a thrombotic microangiopathy and/or a consumptive coagulopathy. Thus further introduction of genes to overcome the coagulation incompatibilities between pig and primate under GalT KO/CRP genetic background has been strongly suggested. CD73 (ecto-5'-nucelotidase) is an enzyme attached via a glycosyl phosphoinositol anchor to the extracellular membrane of endothelial cells, which catalyses the hydrolysis of adenosine triphosphate to adenosine. Loss of activity of CD73 results in activation and aggregation of platelets by a reduced capacity to convert nucleotides to adenosine. In previous study, we reported generation of GalT KO fibroblasts concurrently expressing membrane cofactor protein and produced cloned pigs by nuclear transfer of the fibroblast cells (1). In this study, we constructed a vector for expression of human CD73 under control of promoter of pig Icam2 gene expressed specifically at endothelial cells. This vector was introduced into porcine fibroblasts using the nucleofection technology, by which we had forty three fibroblasts clones carrying pIcam2- CD73 vector. Somatic cell nuclear transfer resulted in generation of two transgenic piglets survived.

Some tissues retain extensive regeneration potential through out adult life and remain as active sites of cell production. Various cell types present in tissues are being produced through proliferation and progressive specialization from a pool of stem cells. In this regard, adult stem cells (ASCs) are multipotent progenitor cells with an ability to proliferate in vitro and undergo extensive self-renewal and differentiation into a wide range of cell types, including adipocytes, chondrocytes, osteocytes, myocytes, cardiomyocytes and neurons. In addition, recent studies showing the abilities of ASCs in generating oocytes-like cells (OLCs) present new perspectives to understand the specification and interaction during the germ cell formation and oogenesis. In the present study, ASCs were established from skin, adipose and ovarian tissues of minipigs. Isolated cells exhibited a fibroblast-like morphology with higher proliferation potential and stronger alkaline phosphatase (AP) activity. ASCs from all tissues expressed pluripotent transcriptional factors, such as Oct-3/4, Nanog and Sox-2 and phenotypic markers, including CD29, CD44, CD90 and vimentin. Further, ASCs were successfully dIfferentiated into osteocytes, adipocytes and neuron-like cells. Upon induction in oogenesis specific media, all ASCs were capable of differentiation into OLCs by exhibiting distinct morphological features. Generated OLCs expressed a range of germ cell specific markers, such as Vasa, deleted in Azoospermia-like (DAZL) factor, stella, c-kit, c-Mos, synaptonemal complex protein 3 (SCP-3), growth differentiation factor 9b (GDF- 9b), zona pellucida C (ZPC) and follicle stimulating hormone receptor (FSHR) at different time points of induction. Differentiated OLCs were also positive for the expression of Vasa and DAZL protein markers. Our findings showing that OLCs can be generated from ASCs of different tissue origin may offer pig as a suitable model for designing transgenic application strategies for reproductive tissue therapy. However, further studies are needed to understand the cellular and molecular mechanisms involved in germ cell differentiation from tissue specific stem cells.

Rhynchium brunneum is a widely distributed wasp species in South Eastern Asia. R. brunneum females were collected from rural provinces of Cambodia, and their total RNA and venom were extracted on site. To search for novel substances in venom, a subtracted cDNA library specific to the venom gland and sac was constructed. A total of 1118 expressed sequenced sequence tags (ESTs) were sequenced and assembled into 349 contigs (107 multiple sequences and 242 singletons). In this result, we found the putative neurotoxin (DTX protein precursor), antimicrobial peptides (teratocyte-specific caboxylesterase) together with typical major components of wasp venom (venom hyaluronidase, arginine kinase, phospholipase A2, serine/theonine protein phosphatase). Additional in-depth annotation would be required for further characterization of many unidentified genes found in the EST library.

2008년 3월부터 10월까지 총 7회에 걸쳐 주왕산국립공원 일대의 식물상을 조사하였다. 주왕산국립공원지역에서 확인된 환경부 지정 멸종위기야생식물종은 둥근잎꿩의비름, 망개나무, 깽깽이풀, 노랑무늬붓꽃과 가시오갈피나무, 솔나리, 연잎꿩의다리 7분류군 이었으며, 식물구계학적 특정식물종은 총 88분류군으로 V등급에 해당하는 분류군은 깽깽이풀, 망개나무, 솔나리 등 8과 9분류군, IV등급에 해당하는 분류군은 승마, 회리바람꽃, 돌마타리 등의 4과 5분류군, III등급에 해당하는 분류군은 애기석위, 가침박달, 등칡 등 16과 18분류군, 2등급으로는 백작약, 나도바람꽃, 개불알꽃 등의 14과 18분류군, 1등급은 늦고사리삼, 홀아비꽃대, 백선 등 30과 38분류군이 확인되었다. 한국 고유 식물종은 호랑버들, 검팽나무, 할미밀망, 노랑갈퀴, 정영엉겅퀴 등의 총 15분류군이 확인되었다. 귀화식물은 닭의덩굴, 나도닭의덩굴, 다닥냉이, 족제비싸리, 컴프리, 수박풀 등의 총 21분류군으로 조사되었다.