• Journal of Korean Neurosurgical Society
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• 제46권5호
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• pp.479-483
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• 2009

Objective : A mouse model of spinal cord injury (SCI) could further increase our basic understanding of the mechanisms involved in injury and repair of the nervous system. The purpose of this study was to investigate whether methods used to produce and evaluate photochemical graded ischemic SCI in rats, could be successfully adapted to mice, in a reliable and reproducible manner. Methods : Thirty female imprinting control region mice (weighting 25-30 g, 8 weeks of age) were used in this study. Following intraperitoneal injection of Rose bengal, the translucent dorsal surface of the T8-T9 vertebral laminae of the mice were illuminated with a fiber optic bundle of a cold light source. The mice were divided into three groups; Group 1 (20 mg/kg Rose bengal, 5 minutes illumination), Group 2 (20 mg/kg Rose bengal, 10 minutes illumination), and Group 3 (40 mg/kg Rose bengal, 10 minutes illumination). The locomotor function, according to the Basso-Beattie-Bresnahan scale, was assessed at three days after the injury and then once per week for four weeks. The animals were sacrificed at 28 days after the injury, and the histopathology of the lesions was assessed. Results : The mice in group 1 had no hindlimb movement until seven days after the injury. Most mice had later recovery with movement in more than two joints at 28 days after injury. There was limited recovery of one joint, with only slight movement, for the mice in groups 2 and 3. The histopathology showed that the mice in group 1 had a cystic cavity involving the dorsal and partial involvement of the dorsolateral funiculi. A larger cavity, involving the dorsal, dorsolateral funiculi and the gray matter of the dorsal and ventral horns was found in group 2. In group 3, most of the spinal cord was destroyed and only a thin rim of tissue remained. Conclusion : The results of this study show that the photochemical graded ischemic SCI model. described in rats, can be successfully adapted to mice, in a reliable and reproducible manner. The functional deficits are correlated an increase in the irradiation time and, therefore, to the severity of the injury. The photothrombotic model of SCI, in mice with 20 mg/kg Rose bengal for 5 minutes illumination, provides an effective model that could be used in future research. This photochemical model can be used for investigating secondary responses associated with traumatic SCI.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.46-46
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• 2003

Interleukin-10 (IL-10) is a homodimeric protein with a wide spectrum of anti-inflammatory and immune activities. It inhibits cytokine production and expression of immune surface molecules in various cell types. The transgenic mice carrying the human IL-10 gene in conjunction with the bovine $\beta$-casein promoter produced the human IL-10 in milk during lactation. Transgenic mice were generated using a standard method as described previously. To screen transgenic mice, PCR was carried out using chromosomal DNA extracted from tail or toe tissues with a primer set. In this study, stability of germ line transmission and expression of IL-10 gene integrated into host chromosome were monitored up to generation F15 of a transgenic line. When female mouse of generation F9 was crossbred with normal male, generation F9 to F15 mice showed similar transmission rates (66.0$\pm$20.13%, 61.5$\pm$16.66%, 41.1$\pm$8.40%, 40.7$\pm$20.34%, 61.3$\pm$10.75%, 49.2$\pm$18.82%, and 43.8$\pm$25.91%, respectively), implying that the IL-10 gene can be transmitted stably up to long term generation in the transgenic mice. For ELISA analysis, IL-10 expression levels were determined with an hIL-10 ELISA and a mIL-10 ELISA kit in accordance with the supplier's protocol. Expression levels of human IL-10 from milk of generation F9 to F13 mice were 3.6$\pm$1.20 mg/ml, 4.2$\pm$0.93 mg/ml, 5.7$\pm$1.46 mg/ml, 6.3$\pm$3.46 mg/ml, and 6.8$\pm$4.52 mg/ml, respectively. These expression levels are higher than in generation F1 (1.6 mg/ml) mice. We concluded that transgenic mice faithfully passed the transgene on their progeny and successively secreted target proteins into their milk through several generations, although there was a little fluctuation in the transmission frequency and expression level between the generations.

• 한국작물학회지
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• 제52권2호
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• pp.146-152
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• 2007

국내육성 벼 주요품종 중 오봉벼, 일품벼 및 아랑향찰벼 과피의 단회 경구투여에 의한 독성을 관찰하기 위하여, 동물체중 당 2.5 g, 5.0 g 및 10.0 g의 용량으로 암수 각각 10마리씩 ICR계 마우스에 1회 경구투여한 후 14일간 실험동물의 사망률, 일반증상, 체중변화, 부검소견 및 혈액 생화학적 분석을 수행하였다. 1. 모든 시험물질의 최대 투여가능용량인 10.0 g/kg 투여시 암수 모든 동물군의 사망 예는 관찰되지 않았고, 또한 그 이하의 시료 투여군에서도 사망동물은 없었으며, 시험물질 투여 후 나타내는 외관상의 이상증상도 확인되지 않았다. 2. 실험동물의 체중 변화는 암수 모두에서 시일이 경과함에 따라 체중 증가가 일시적으로 억제되는 경향을 나타냈으나 시험물질 투여군과 대조군간의 유의성 있는 차이는 나타나지 않았다. 3. 시료투여 14일 후 치사된 동물의 장기를 관찰한 결과, 암수 모든 실험군의 간장, 신장, 비장, 심장, 폐 및 뇌에서 시험물질 투여에 따른 어떤 육안적 이상 소견도 발견되지 않았다. 4. 혈액생화학적 검사 결과, ALT와 AST 활성도가 모든시료 투여군에서 정상수치를 나타냈으며, 10.0 g/kg 1회 경구 투여한 모든 실험군의 경우에도 정상 수치를 나타내는 것으로 보아, 고품질 우리 쌀인 오봉벼, 일품벼 및 아랑향찰 벼 과피 추출물을 고용량으로 투여 시에도 간 기능에 어떠한 독성도 나타내지 않음을 확인할 수 있었다. 5. 오봉벼, 일품벼 및 아랑향찰벼 과피 추출물에 대한 급성 경구 독성시험에서 상기의 일반상태, 체중변화 및 부검소견 등에 별다른 독성이 관찰되지 않았으며, 최대투여 가능용량인 10.0 g/kg에서도 사망 예가 발견되지 않았다. 따라서 암수 마우스에 대한 경구 $LD_{50}$치는 최대 투여가능 용량인 10.0 g/kg 이상으로 평가되었다. 따라서 본 시험 결과, 오봉벼, 일품벼 및 아랑향찰벼 과피 EtOH 추출물은 투여가능 최대용량에서도 독성이 없는 안전한 식품임을 확인하였고, 건강증진에 도움을 주는 기능성 식품소재로서의 개발 가능성을 예측할 수 있었으며, 고용량으로도 임상사용이 가능함을 확인할 수 있었다.

• IMMUNE NETWORK
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• 제4권1호
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• pp.53-59
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• 2004

Background: Minor histocompatibility HY antigen, as a transplantation antigen, has been known to cause graft rejection in MHC (major histocompatibility complex) matched donor-recipient. The aim of our study is to investigate the role of male antigen (HY) disparity on MHC matched pancreatic islet transplantation and to examine the mechanism of the immune reaction. Methods: Pancreatic islets were isolated and purified by collagen digestion followed by Ficoll gradient. The isolated islets of male C57BL6/J were transplanted underneath the kidney capsule of syngeneic female mice rendered diabetic with streptozotocine. Blood glucose was monitored for the rejection of engrafted islets. After certain period of time, tail to flank skin transplantation was performed either on mouse transplanted with HY mismatched islets or on sham treated mouse. The rejection was monitored by scoring gross pathology of the engrafted skin. Results: HY mismatched islets survived more than 300 days in 14 out of 15 mice. The acceptance of second party graft (male B6 islets) and the rejection of third party graft (male BALB/c islets) in these mice suggested the tolerance to islets with HY disparity. B6 Skin with HY disparity was rejected on day $25{\pm}7$. However, HY mismatched skin transplanted on the mice tolerated to HY mismatched islets survived more than 240 days. Tetramer staining in these mice indicated the CTL recognizing MHC Db/Uty was not deleted or anergized. Conclusion: The islet transplantation across HY disparity induced tolerance to HY antigen in C57BL6 mouse, which in turn induced tolerance to HY mismatched skin, which otherwise would be rejected within 25 days. The MHC tetramer staining suggested the underlying mechanisms would not be clonal deletion or anergy.

• Parasites, Hosts and Diseases
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• 제52권2호
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• pp.151-162
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• 2014

The technique of stem cells or hepatocytes transplantation has recently improved in order to bridge the time before whole-organ liver transplantation. In the present study, unfractionated bone marrow stem cells (BMSCs) were harvested from the tibial and femoral marrow compartments of male mice, which were cultured in Dulbecco's modified Eagle's medium (DMEM) with and without hepatocyte growth factor (HGF), and then transplanted into Schistosoma mansoni- infected female mice on their 8th week post-infection. Mice were sacrificed monthly until the third month of bone marrow transplantation, serum was collected, and albumin concentration, ALT, AST, and alkaline phosphatase (ALP) activities were assayed. On the other hand, immunohistopathological and immunohistochemical changes of granuloma size and number, collagen content, and cells expressing OV-6 were detected for identification of liver fibrosis. BMSCs were shown to differentiate into hepatocyte-like cells. Serum ALT, AST, and ALP were markedly reduced in the group of mice treated with BMSCs than in the untreated control group. Also, granuloma showed a marked decrease in size and number as compared to the BMSCs untreated group. Collagen content showed marked decrease after the third month of treatment with BMSCs. On the other hand, the expression of OV-6 increased detecting the presence of newly formed hepatocytes after BMSCs treatment. BMSCs with or without HGF infusion significantly enhanced hepatic regeneration in S. mansoni-induced fibrotic liver model and have pathologic and immunohistopathologic therapeutic effects. Also, this new therapeutic trend could generate new hepatocytes to improve the overall liver functions.

• 한국식품영양과학회지
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• 제31권1호
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• pp.155-159
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• 2002

AEY의 acetone 추출물이 mouse liver microsome의 산화에 미치는 영향을 조사하였다. Mouse liver microsome에 함유된 단백질 AEY 처리가 CEY 처리구에 비해 상대적으로 7~48%정도가 증가하였다. 지방산 분석 결과, CEY 처리에 비해 AEY 처리구에서 stearic acid를 포함하는 포화지방산의 함량비가 상대적으로 높았으며, oleic acid, linoleic acid 등을 함유하는 불포화지방산의 함량비가 현저히 감소하였다. AEY 처리구는 F $e^{2+}$가 관여하는 Asc/F $e^{2+}$, NADPH/F $e^{+2}$ 를 산화유도물질로 사용한 경우에 TC와 비슷하거나 보다 강력한 항산화 효과를 보였다. ABIN이나 CuOOH가 산화유도물질로 작용할 시에는 AEY처리에 의해 유의성있는 항산화 효과가 나타나지 않았다.

• Toxicological Research
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• 제11권1호
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• pp.15-21
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• 1995

Several populations of lymphocytes possess receptors for autonomic neurotransmitter, which make lymphocytes susceptible to autonomic stimulation. This study was to evaluate the functional alternation of effector cells of the immune system. Female Balb/C mice, 15-20 g, were injected with MA subcutaneously under various conditions. Mixed lymphocyte reaction (MLR) showed certain T cell subsets were affected by MA. The level of interleukin-2 (IL-2) production was inhibited due to a defect in expression of the IL-2 receptor. In mice injected with 20 mg MA/kg, 1 day before assay, phagocytosis of peritoneal macrophages showed $14.07\pm3%$, which was similar degree to 5 mg MA/kg treatment for 4 consecutive days. Phagocytosis was almost recovered to that of control after 4 day in 20 mg/kg injected mice. Maximum inhibition of plaque forming cell (PFC) occurred when MA was given early, indicating the inductive time point of antibody production was affected. The cortisol level increased in the MA treated group (0.05, 0.20, and $0.08{\mu}g$/dl for control, low, and high dose-MA treated mice, respectively). Based on these results, MA has general suppression effects on the immune systems by functional alteration of effector cells. Considering the increment of serum cortisol levels, MA partially impacts the neuroendocrine system to lead to failure of immune response.

• Toxicological Research
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• 제23권1호
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• pp.65-72
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• 2007

This study was conducted to obtain information of the oral dose acute toxicity of PGB-2, a novel polyglucosamine polymer produced from Citrobacter sp. BL-4 (a new strain) in male and female mice. Mortality, body weight changes, clinical signs were monitored during 14 days after single oral dose of test article at dose levels of 2000, 1000, 500, 250 and 125 ml/kg. Gross lesions, organ weight and histopathology of principal organs were examined after necropsy. As the results, we could not find any mortalities, clinical signs, changes in the body weight and gross findings except for white foci in the liver. In addition, no PGB-2-treatment related abnormal changes on the organ weight and histopathology of principle organs were detected except for atypical signs of liver. White liver foci were confirmed as focal infiltration of inflammatory cells. The results suggest that the PGB-2 is relatively safe in mice but the possibility of hepatotoxicity could not be excluded. The $LD_{50}$ and approximate LD in mice after single oral dose of PGB-2 were considered over 2000 mg/kg, respectively. In future, the potential hepatotoxicity of PGB-2 should be evaluated through the repeat dose toxicity test prior to develop as a new agent.

• 대한한의학방제학회지
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• 제12권2호
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• pp.95-108
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• 2004

Objective: We made the genital organs inflammatory mice model by vaginal injection of Lipopolysaccharide(LPS), and we intended to make study about anti-inflammatory effects of Gangyeong-Tang, among oral and rectal medication and Herbal-Acupuncture treatment. Method: The female ICR(20-30g) mice were used, the temperature was controlled within $22{\pm}0.5^{\circ}C$ and water and food was not limited. The environment was manipulated to simulate 12 hours of day and 12 hours of night. After LPS injection into vagina, we confirmed on-set of inflammation. 2 days before LPS injection, we started to medicate Gangyeong-Tang in Oral and Rectal and Aqua-acupucture treatment. After 3days from LPS injection, we mesured the White Blood Cell(WBC), Interrleukin-6(IL-6), Tumor Necrosis Factor-${\alpha}$(TNF-${\alpha}$) in blood which was collected from the Retro-orbital Plexus. Results: 1. We made the genital organs inflammatory mice model by vaginal injection of LPS successfully. 2. The number of WBC was decreased significantly as we medicated Gangyeong- Tang in Oral 1g/kg dose, 3g/kg dose and Rectal 1g/kg dose, rectal 3g/kg dose. 3. The concentration of IL-6 was decreased significantly as we medicated Gangyeong-Tang in all group of the Oral, Rectal, Herbal-Acupuncture treatment. 4. The concentration of TNF-${\alpha}$ was decreased significantly as we medicated Gangyeong-Tang in Oral 3g/kg dose and Rectal 1g/kg dose, Rectal 3g/kg dose group. Herbal-Acupuncture treatment group datas showed reductive tendency. Conclusion: As a result of this experiment, we made the genital organs inflammatory mice model by vaginal injection of LPS successfully and demonstrated anti-inflammatory effect of Gangyeong-Tang.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.17-17
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• 2003

The milk of transgenic animals may provide an attractive vehicle for large-scale production of hEPO. Since glycosylation is cell type specific, recombinant human EPO (rhEPO) produced in different host cells contain different patterns of oligosaccharides, which could affect the biological functions. However, there have been no reports on the characteristics of rhEPO derived from milk of transgenic animals. To address this objective, several transgenic mice by using pWAPhEPO and/or pBC1hEPO expression vector were produced. However, 2 lines of pWAPhEPO founder female mouse died during late gestational day (day 18) before offspring could be obtained. They showed a severe splenomegaly, Unlike those of pWAPhEPO, mammary gland epithelial cells from biopsies of lactating pBC1hEPO transgenic mice had marked immunoreactivity to EPO and any activity was not detected in other tissues. The expression level of rhEPO is about 0.7% of mammary gland cellular total soluble proteins and an amount of 300~500 mg/L rhEPO is secreted into milk. Furthermore, the pBC1hEPO transgenic mice transmitted this character to their progeny in mendelian manner. In order to determine the extent of glycosylation variation, N-linked oligosaccharide structures present in the milk-derived rhEPO were characterized. Most of milk-derived rhEPO is fully glycosylated. the biological activity of milk-derived rhEPO was comparable to that of purified CHO-derived rhEPO, and milk-derived rhEPO showed relatively stable after freezing and thawing. Taken together, the results illustrate the potential of transgenic animals in the large-scale production of biopharmaceuticals.