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A Study the Effects of Three Preparations of Hirudo on the Expression of Pro-inflammatory Cytokines in Human Bronchial Epithelial Cells Line BEAS-2B (수질(水蛭)의 제법(制法)에 따른 BEAS-2B 인간(人間) 기관지상피세포(氣管支上皮細胞)의 염증유발성(炎症誘發性) Cytokines 발현(發顯)에 미치는 영향(影響))

  • Jung, Hee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo;Han, Dong-Ha
    • The Journal of Internal Korean Medicine
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    • v.25 no.4
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    • pp.260-273
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    • 2004
  • Backgrounds : In recent years, asthma has become recognized as a chronic inflammatory disease associated pathologically with airway epithelial inflammation and airway remodeling. Objectives : To evaluate the different effects of Hirudo depending upon pharmaceutical manufactures on the expression and the activities of IL-6 and GM-CSF in airway epithelial cells, samples of Hirudo(水蛭), Hirudo toasted with Talcum(水蛭滑石炒) and Hirudo toasted with Ephedrae Herba(水蛭麻黃炒) were tested. Methods : After inducing enhanced messenger RNA(mRNA) expression and secretion of each cytokine by tumor necrosis factor-alpha(10 ng/ml) treatment, cultured human bronchial epithelial cell line BEAS-2B was added to each sample$(l,\;10,\;100\;&\;1000\;{\mu}g/ml)$. Subsequently, DNA activities were analyzed. Specifically mRNA expression and culture supernatants(protein levels) of IL-6 and GM-CSF from BEAS-2B cells, were analyzed using luciferase reporter gene assay, reverse transcription-polymerase chain reaction(RT-PCR) analysis and enzyme-linked immunosorbent assay. Results : Hirudo toasted with Ephedrae Herba(水蛭麻黃炒) and Hirudo(水蛭) inhibited IL-6 activities in BEAS-2B cells remarkably, and inhibited mRNA expression levels and protein levels in supernatant of IL-6 and GM-CSF at various concentrations, significantly(p<0.05). However, Hirudo toasted with Talcum(水蛭滑石炒) had no effect on mRNA expression levels and showed a slight inhibitory effect on GM-CSF protein levels in supernatant of culture medium. Conclusions : These results strongly suggest that Hirudo toasted with Ephedrae Herba(水蛭麻黃炒) and Hirudo(水蛭) would be serve as effective medicaments in the treatment of airway inflammation and remodeling of asthmatic patients.

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Characterization of Transgenic Tall Fescue Plants Expressing Two Antioxidant Genes in Response to Environmental Stresses (두 가지 항산화유전자를 동시에 발현시킨 형질전환 톨 페스큐 식물체의 환경스트레스에 대한 내성 특성 해명)

  • Lee, Sang-Hoon;Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Seo, Sung;Kwak, Sang-Soo;Kwon, Suk-Yoon;Yun, Dae-Jin;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.2
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    • pp.109-116
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    • 2007
  • Environmental stress is the major limiting factor in plant productivity. As an effort to solve the global food and environmental problems using the plant biotechnology, we have developed transgenic tall fescue (Festuca arundinacea Schreb.) plants via Agrobacterium-mediated gene transfer method. To develop transgenic tall fescue plants with enhanced tolerance to the environmental stresses, both CuZn superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) genes were incorporated in a pIG121 binary vector and the both of the genes were controlled separately by an oxidative stress-inducible sweet potato peroxidase 2 (SWPA2) premoter expressed in chloroplasts. Leaf discs of transgenic plants showed 10-30% less damage compared to the wild-type when they exposed to a wide range of environmental stresses including methyl viologen (MV), $H_2O_2$ and heavy metals. In addition, when $200{\mu}M$ MV was sprayed onto the whole plants, transgenic plants showed a significant reduction of visible damage compared to wild-type plants that were almost damaged. These results suggest that over expression of CuZnSOD and APX genes in transgenic plants might be a useful strategy to protect the crops against a wide range of environmental stresses.

Antibacterial Activity and Probiotic Potential of Lactobacillus plantarum HKN01: A New Insight into the Morphological Changes of Antibacterial Compound-Treated Escherichia coli by Electron Microscopy

  • Sharafi, Hakimeh;Maleki, Hadi;Ahmadian, Gholamreza;Zahiri, Hossein Shahbani;Sajedinejad, Neda;Houshmand, Behzad;Vali, Hojatollah;Noghabi, Kambiz Akbari
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.225-236
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    • 2013
  • Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.

Profiles of Bacillus spp. Isolated from the Rhizosphere of Suaeda glauca and Their Potential to Promote Plant Growth and Suppress Fungal Phytopathogens

  • Lu, Ping;Jiang, Ke;Hao, Ya-Qiao;Chu, Wan-Ying;Xu, Yu-Dong;Yang, Jia-Yao;Chen, Jia-Le;Zeng, Guo-Hong;Gu, Zhou-Hang;Zhao, Hong-Xin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1231-1240
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    • 2021
  • Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

Clinical and Genetic Features of Korean Inherited Arrhythmia Probands

  • Joo Hee Jeong;Suk-Kyu Oh;Yun Gi Kim;Yun Young Choi;Hyoung Seok Lee;Jaemin Shim;Yae Min Park;Jun-Hyung Kim;Yong-Seog Oh;Nam-Ho Kim;Hui-Nam Pak;Young Keun On;Hyung Wook Park;Gyo-Seung Hwang;Dae-Kyeong Kim;Young-Ah Park;Hyoung-Seob Park;Yongkeun Cho;Seil Oh;Jong-Il Choi;Young-Hoon Kim
    • Korean Circulation Journal
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    • v.53 no.10
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    • pp.693-707
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    • 2023
  • Background and Objectives: Inherited arrhythmia (IA) is a more common cause of sudden cardiac death in Asian population, but little is known about the genetic background of Asian IA probands. We aimed to investigate the clinical characteristics and analyze the genetic underpinnings of IA in a Korean cohort. Methods: This study was conducted in a multicenter cohort of the Korean IA Registry from 2014 to 2017. Genetic testing was performed using a next-generation sequencing panel including 174 causative genes of cardiovascular disease. Results: Among the 265 IA probands, idiopathic ventricular fibrillation (IVF) and Brugada Syndrome (BrS) was the most prevalent diseases (96 and 95 cases respectively), followed by long QT syndrome (LQTS, n=54). Two-hundred-sixteen probands underwent genetic testing, and 69 probands (31.9%) were detected with genetic variant, with yield of pathogenic or likely pathogenic variant as 6.4%. Left ventricular ejection fraction was significantly lower in genotype positive probands (54.7±11.3 vs. 59.3±9.2%, p=0.005). IVF probands showed highest yield of positive genotype (54.0%), followed by LQTS (23.8%), and BrS (19.5%). Conclusions: There were significant differences in clinical characteristics and genetic yields among BrS, LQTS, and IVF. Genetic testing did not provide better yield for BrS and LQTS. On the other hand, in IVF, genetic testing using multiple gene panel might enable the molecular diagnosis of concealed genotype, which may alter future clinical diagnosis and management strategies.

THE COMPARISON OF STREPTOCOCCUS MUTANS ISOLATED FROM OCCLUSAL SURFACES OF CARIES AND NON-CARIES TEETH (우식치아와 정상치아의 교합면에서 분리한 Streptococcus mutans의 비교)

  • Park, Ho-Won;Jung, Tae-Sung;Jung, Jin;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.1
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    • pp.129-141
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    • 2001
  • When oral microorganisms were sampled from occlusal surfaces of caries and non-caries teeth, $3.43\times10^5$ CFU and $3.47\times10^3$ CFU of bacteria were counted on MSB agar plates, respectively. All the 20 colonies isolated from a caries surface were Streptococcus mutans but, only two of 20 colonies were identified as Streptococcus mutans by API test. S. mutans SM1 from caries tooth and S. mutans SM2 from non-caries tooth showed the same results except for $\alpha-galactosidase$ activity on sugar fermentation tests and biochemical tests. For the bacterial replication, both SM1 and SM2 were actively multiplicated at pH 5.5. And the viability of SM1 was high at 20% of sucrose, while that of SM2 was high at 5% of sucrose in the media. SM1 actively replicated at 16mM of $CaCl_2$, 160mM of KCl, and 6.4mM of $MgCl_2$, and the replication of SM2 was increased at 16mM of $CaCl_2$, 40mM of KCl, 6.4mM of $MgCl_2$. At 1mM of sodium bicarbonate and sodium phosphate, both bacteria were actively multiplicated. SM1 and SM2 were actively replicated at 1mM and 10mM of Tris, respectively. For potassium phosphate buffer, SM1 grew well proportionally to the concentration up to 100mM, while the growth of SM2 were inhibited by the increase of concentration. The 4.6 kb of gtf gene was amplified with a pair of primer, gtfB-F961 and gtfC-R5574 by polymerase chain reaction from the chromosomal DNA of SM1 and SM2. When 4.6kb bands were eluted from gel and were treated with restriction enzyme, EcoR I produced the same RFLP like 0.8kb and 3.8kb of DNA fragments for S. mutans GS-5, SM1 and SM2. By Hind III, the PCR products weren't digested for S. mutans GS-5 and SM1, but 3 fragments such as 2.4kb, 1.8kb and 400bp were examined for SM2. These results indicated the difference between gtf genes of SM1 and SM2. BamH I treatment showed 4 fragments for SM1 and SM2, while the 3 fragments for S. mutans GS-5. The PCR products were not digested by Kpn I, Sma I, Xho I and Pst I.

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Effects of Vitamin $K_1$ on the Developmental and Survival Rate of Porcine In Vitro Fertilized Embryos (Vitamin $K_1$의 첨가가 돼지 체외 수정란의 발달과 생존율에 미치는 효과)

  • Park, Hum-Dai;Zhu, Yi-Chen;Park, Yong-Soo
    • Journal of Embryo Transfer
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    • v.29 no.1
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    • pp.73-81
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    • 2014
  • The in vitro production of porcine embryos was essential to increase of blastocyst development rate and select of high quality blastocyst in early stage. There were a lot of reports about in vitro porcine embryo development, but there was no report about the selection of high quality embryos. Therefore, in this study, we investigated the effect of vitamin $K_1$ (vit $K_1$) on the development and survival rate of porcine in vitro fertilized embryos. When vit $K_1$ was treated for 24 hr at day 1 in vitro culture, blastocyst development rate in the control group ($35.5{\pm}3.2%$) was significantly lower compared to $1.0{\mu}M$, $3.0{\mu}M$, or $6.0{\mu}M$ groups ($14.5{\pm}4.3$, 0.0, or 0.0%; p<0.05). The survival rates of blastocysts at day 8 in $1.0{\mu}M$, $3.0{\mu}M$ or $6.0{\mu}M$ of vit $K_1$ treated groups ($22.2{\pm}2.9$, 0.0 or 0.0%) were significantly lower than that of the control group ($31.8{\pm}2.6%$; p<0.05). We were added at $1.0{\mu}M$, $3.0{\mu}M$ or $6.0{\mu}M$ vit $K_1$ for different durations of time at day 1 in vitro culture. The development rate and survival rate in the group of $1.0{\mu}M$ vit $K_1$ for 6 hr was $26.5{\pm}2.9%$ and $47.2{\pm}2.8%$, respectively, which were differed significantly in the group of 12 hr (p<0.05). In the group of $3.0{\mu}M$ vit $K_1$, the blastocyst development in control group was $36.4{\pm}3.1%$ but, the survival rate $41.7{\pm}3.2%$ in the group of 3.0 hr was significantly higher than that of the control group (p<0.05). In the group of $6.0{\mu}M$ vit $K_1$, the control group's the blastocyst development was $32.0{\pm}2.8%$ and the 0.5 hr supplement group's survival rates was $42.9{\pm}1.8%$ higher than other groups. We added vit $K_1$ at day 1, day 2, day 4 and day 6 of in vitro culture, on the based the results of supplemented concentration and duration. In the group of $1.0{\mu}M$ 6.0 hr addition, the blastocyst development rate of day 4 and the survival rate of day 2 were the highest in each group. In the groups of $3.0{\mu}M$ 3.0 hr addition or $6.0{\mu}M$ 0.5 hr addition, the blastocyst development ($59.5{\pm}4.1%$ and $50.0{\pm}3.6%$) and survival rates ($72.7{\pm}5.4%$ and $79.2{\pm}4.0%$) on day 4 were significantly higher than that of control and other experiment groups (p<0.05). Meanwhile, the number of cells in blastocysts that produced by vit $K_1$ supplementation was $53.4{\pm}5.8$, $49.4{\pm}3.8$ and $51.5{\pm}4.5$ respectively, which were significantly higher than that of $40.2{\pm}2.3$ in the control group (p<0.05). There was no difference of the number of apoptotic cells between control and experiment groups. In addition, gene expression of survival blastocyst, the Bax mRNA expression was similar between the control and the experiment groups. However, Bcl-xL mRNA expression's in the group of $6.0{\mu}M$ 0.5 hr on day 4 was highest among control and experiment groups (p<0.05). In this study suggested that the control of concentration, duration and time was effective on the survival and cell number of porcine blastocyst derived from in vitro. We are not know what the exact reasons of the effect of vit $K_1$ on embryo development and need to fur ther study. However, vit $K_1$ might be using the selection of high quality porcine blastocyst.

Expression Analysis of Anthocyanin Biosynthetic Genes of Tassel and Silks in Gwangpyeongok and Dacheongok (광평옥과 다청옥의 수이삭과 수염에서 안토시아닌 생합성 유전자 발현 분석)

  • Go, Young Sam;Bae, Hwan Hee;Choi, Yu Chan;Son, Jae Han;Ha, Jun Young;Shin, Seong Hyu;Jung, Tae Wook
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.3
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    • pp.240-247
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    • 2021
  • Anthocyanins are known to be involved in various functions such as antioxidant and antibacterial activities in plants. Although studies on anthocyanins in corn have been conducted recently, basic research related to anthocyanin biosynthesis is insufficient. In this study, we examined the molecular biological and physicochemical properties related to anthocyanin biosynthesis in the tassel and silks of Gwangpyeongok and Dacheongok cultivars. Anthocyanins were not synthesized in either the tassel or silks in Gwangpyeongok, whereas were synthesized in both in Dacheongok. The total anthocyanin content was approximately 30 times higher in the tassel and silks of Dacheongok than in those of Gwangpyeongok. In addition, C-3-G was measured only in the tassel of Dacheongok, and C-3-G, Pg-3-G, and M-3-G were 45.2 times, 27.3 times, and 37.6 times higher, respectively, in the silks of Dacheongok than of Gwangpyeongok. Expression of F3'H, DFR, and GST genes decreased in the tassel, and that of F3'H and DFR genes decreased in the silks of Gwangpyeongok. It was further confirmed that transcription factor P1 and R1 regulate the expression of anthocyanin biosynthetic genes in the tassel and silks, respectively, in Gwangpyeongok. Linoleic acid (C18:2) decreased by 6.6% and 10.9%, and linolenic acid (C18:3) increased by 8.5% and 8.5%, in the tassel and silks, respectively, of Gwangpyeongok compared to those of Dacheongok. Palmitic acid (C16:0) increased by 4.1% and oleic acid (C18:1) decreased by 2.1% in the silks of Gwangpyeongok compared to that in Dacheongok. In addition, the total fatty acid content in the tassel and silks increased by 10.3% and 30.4%, respectively, in Gwangpyeongok compared to that in Dacheongok. However, no significant results were observed in the analysis of phytosterol components. These results may be utilized as useful resources for the development of functional corn containing a large amount of anthocyanins.

Enhanced biological effect of fermented soy-powder milk with Lactobacillus brevis increasing in γ-aminobutyric acid and isoflavone aglycone contents (가바와 비당체 이소플라본이 증가된 Lactobacillus brevis 발효 콩-분말 두유의 생리활성 증진 효과)

  • Hwang, Chung Eun;Kim, Su Cheol;Lee, Jin Hwan;Hong, Su Young;Cho, Kye Man
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.245-255
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    • 2018
  • The research was aimed to analyze the functional constituents (GABA and isoflavone), radical (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and hydroxyl) scavenging activities and enzyme (${\alpha}-glucosidase$ and lipase) inhibitory effects of soypowder milk (SPM) and fermented soy-powder milk (FSPM) with varied Lactobacillus brevis. Ten ${\gamma}-aminobutyric$ acid (GABA) producing lactic acid bacteria that showed 96-99% similarity with L. brevis, according to 16S rRNA gene sequence analysis, were isolated from fermented kimchi. The conversion rates of GABA were obtained 66.96-93.51, 63.76-84.58, and 57.05-69.75% in monosodium glutamate, gluten and soy protein, respectively. The levels of pH and glutamic acid of FSPM were found lower than those of SPM, but the acidity and GABA contents were higher. The GABA conversion rate of FSPM with BMK484 strain was attained the highest 69.97%. The contents of isoflavone glycoside ($1290.93{\mu}g/g$) was higher in SPM, but the content of isoflavone aglycone ($287.27-501.9{\mu}g/g$) was higher in FSPM. The levels of isoflavone aglycone such as daidzein, glycitein and genistein, were found as the highest 240.2, 61.24 and $200.45{\mu}g/g$, respectively, when FSPM was made with BMK484 strain. The DPPH, ABTS and hydroxyl radical scavenging and ${\alpha}-glucosidase$ and pancreatic lipase inhibitory activities of FSPM made with BMK484 strain were the relatively higher 60.31, 88.10, 61.25, 52.71, and 39.37%, respectively. Therefore, the L. brevis can be used as a material capable of simultaneously enhanced GABA and isoflavone aglycone in FSPM.

Stable Transmission and Continuous Expression of Human Interleukin-10 Transgene in the Offspring of Transgenic Mice (형질전환 생쥐의 후대에서 인간 Interleukin-10 유전자의 안정적 전이와 지속적인 발현)

  • Zheng Z. Y.;Koo D. B.;Han Y. M.;Lee K. K.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.203-207
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    • 2004
  • The transgenic mice carrying human Interleukin-10 (hIL-10) gene in conjunction with bovine (3 -casein promoter express hIL-10 in milk during lactation. In this study, stability of germ line transmission and expression of hIL-10 transgene integrated into host chromosome were monitored up to generation F8 of transgenic mice. When male mouse of generation F8 was crossbred with normal females, approximately half of offspring (50.9±5.8%) were identified as transgenic mice. Generation F9 to F15 mice also showed similar transmission rates (66.0±20.1%, 61.5±16.7%, 41.1±8.4%, 40.7±20.3%, 61.3±10.8%, 49.2±18.8% and 43.8±25.9%, respectively), implying that hIL-10 transgene can be transmitted stably up to long term generation in the transgenic mice. Expression levels of human IL-10 from milk of generation F9 to F14 mice were 3.6± 1.2 mg/ml, 4.2±0.9 mg/ml, 5.7±1.5 mg/ml, 6.3±3.5 mg/ml, 6.8±4.5 mg/ml and 6.8±3.1 mg/ml, respectively, which was showed high-level expression compared with that of generation F1 (1.6 mg/ml) mice. In conclusion, our results suggest that transgenic mice can be continuously passed their transgenes to the progeny through the breeding program with the same productivity of human IL-10 protein in their milk.