• Journal of Microbiology and Biotechnology
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• 제32권10호
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• pp.1335-1343
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• 2022

COVID-19 is an emerging disease that poses a severe threat to global public health. As such, there is an urgent demand for vaccines against SARS-CoV-2, the virus that causes COVID-19. Here, we describe a virus-like nanoparticle candidate vaccine against SARS-CoV-2 produced by an E. coli expression system. The fusion protein of a truncated ORF2-encoded protein of aa 439~608 (p170) from hepatitis E virus CCJD-517 and the receptor-binding domain of the spike protein from SARS-CoV-2 were expressed, purified and characterized. The antigenicity and immunogenicity of p170-RBD were evaluated in vitro and in Kunming mice. Our investigation revealed that p170-RBD self-assembled into approximately 24 nm virus-like particles, which could bind to serum from vaccinated people (p < 0.001) and receptors on cells. Immunization with p170-RBD induced the titer of IgG antibody vaccine increased from 14 days post-immunization and was significantly enhanced after a booster immunization at 28 dpi, ultimately reaching a peak level on 42 dpi with a titer of 4.97 log₁₀. Pseudovirus neutralization tests showed that the candidate vaccine induced a strong neutralizing antibody response in mice. In this research, we demonstrated that p170-RBD possesses strong antigenicity and immunogenicity and could be a potential candidate for use in future SARS-CoV-2 vaccine development.

• 동의생리병리학회지
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• 제21권5호
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• pp.1210-1218
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• 2007

Recently Atopic Dermatitis(AD) is increasing along with allergic disease. At present, there is no infallible cure for AD. Then AD patients undergo great suffering. This study is carried out to see whether or not the administering Danggwieumja(DG) along with Samhwangseje-gamibang(SG} as a medicine for external aplication, is effective in treating atopic dermatitis. To examine the effectiveness of the above prescription, the author made an observation of diverse immune responses. through the model of NC/Nga atopic mice. Results provided evidence that the DG administration along with SG can be used as a treatment means to atopic dermatitis. The results are as follows: The extent of Clinical skin severities in 13 and 16 week old NC/Nga mice treated with DG and SG, were reduced by 50.9%, 53.9% respectively, compared to the control NC/Nga mice with no drug treatment. IgE, IL-4, IL-5, IL-6, IgM and IgG1 levels in the serum of the NC/Nga mice treated with DG and SG were significantly decreased compared to those of the untreated control mice. In contrary, to the $IFN-{\gamma}$ level, significantly increased. The spleen weight of the NC/Nga mice treated with DG and SG significantly decreased compared to those of the untreated control mice. CCR3 gene expression in the skin tissue of NC/Nga mice treated with DG and SG were highly decreased, and the IL-6 expression significantly decreased, and the $IFN-{\gamma}$ gene expression increased compared to those of the untreated control mice. Histological observation of the ear and dorsal skin tissue of the NC/Nga mice treated with DG and SG, showed that the extents of inflammation and infiltrated immune cells in the epidermal tissue and dermis, were highly reduced compared to those of the untreated control mice. In the model inducing COX-2 activity in RAW 264.7 cell, the denser DG became, the more COX-2 activity was inhibited, compared to those of the untreated control group. $IL-1{\beta}$, and $TNF-{\alpha}$, IL-6 gene expression in RAW 264.7 cell with DG, significantly decreased, compared to those of the untreated control group. According to the assessment of cell toxicity in L929 cell, the rate of cell multiplication increased by 3% in consistency to 100ppm of DG compared to the untreated control group and in more than the 200 ppm consistency, cell toxicity was occurred.

• 한방안이비인후피부과학회지
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• 제19권2호
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• pp.1-18
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• 2006

Objective : Although management of asthma has become increasingly effective, its cure remains elusive, necessitating a new modality to prevent or eliminate causes triggering clinical progress. Based in the clinical experiences, a novel decoration Cheongsangbiyeum (CSB), has been developed to treat asthma, which consists of Polyporus, Semen Myristicae, Pericarpium citri Reticulatae, Rhizoma Cimicifugae, Cortex Albizziae, Fructus Rubi, Rhizoma Zedoariae, and Rhizoma Rhei. In the current study, its anti-asthmatic efficacy was evaluated using a mouse model of asthma. Methods : Experimental allergic asthma was induced by repeated intraperitioneal sensitization and intranasal challenge of ovalbumin (OVA). Water extract of CSB (1 mg/mouse/day) was administrated orally whereas control mice on given with identical volume of phosphate-buffered saline (PBS) for 5 days during the course of antigen challenge. When airway hyperreactivity(AHR) measured by ${\bata}-methacoline-induced$ airflow obstruction was compared, AHR of CSB-treated mice was significantly lower than those of control mice, indicating that CM extract can attenuate an asthmatic symptom. Airway recruitment of leukocytes and eosinophils was also markedly reduced by CSB treatment suggesting that oral treatment of CSB can alleviate the airway inflammation. For a better understanding of possible mechanisms underlying anti-asthmatic effet of CSB, cytokine (IL-4, IL-5, IL-13 and $IFN{\gamma}$ levels in bronchoalveola lavage fluid (BALF) and lung tissues were determined. Results : The results showed that cytokine levels were significantly lowered by CSB treatment. Additionally, number of draining lymph node cells was significantly lower than those of control mice. These data indicate that CSB suppress in vivo allergen-specific response. However, notably, levels of type 2 cytokines such as IL-5 and IL-13 were more profoundly influenced. Moreover, in vitro OVA-specific proliferative response and type 2 cytokine (IL-4, IL-5 and IL-13) production lymph node cells was markedly decreased in CSB-treated mice, whereas their $IFN{\gamma}$ production was not significantly altered Thrse data clearly showed a preferential inhibition of type 2 T cell (Th2) response by CSB treatment. This finding was also supported by serum antibody data showing that levels of OVA-specific type 2 antibodies, IgE and IgG1, in CSB-treated mice were significantly lower than in control mice, while type 1 antibody, IgG2a level m rather higher than controls, although the difference was in significant. Conclusions : In conclusion, oral administration of CSB attenuates asthmatic manifestations including AHR ad airway recruitment of eosinophils in a mouse model which possibly results from selective inhibition of Th2 cell response to allergen. Our data suggest a potential clinical application of CSB for control of allergic asthma.

• 한국가금학회지
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• 제33권2호
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• pp.141-149
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• 2006

본 실험은 항생제 $BMD^(R)$ premix(Bacitracin methylene disalicylate), 유기산 복합제인 $Lactacid^(R)$((주) 은진인터내셔날) 그리고 essential oil 제품인 $Immunocin^(R)$((주) 은진인터내셔날) 이 육계에 미치는 영향을 비교 평가하기 위하여 생산성, 영양소 이용율, 혈액 성상, ND titer, IgG, 그리고 장내 미생물 균총 등을 조사하기 위해 실시하였다. 사양 시험은 육계 ($Ross^(R)$종) 1,000수 (암 수 각각 500수)를 공시하여 4처리 5반복, 반복당 암 수 동수로 50수씩을 floor pen(가로: 2.0m, 세로: 2.4m) 에 완전임의 배치하였다. 사양시험 기간은 35일, 시험 기간 동안 물과 사료를 자유 채식시켰고 24시간 점등하였다. 시험구는 대조구, $BMD^(R)$ premix(Bacitracin methylene disalicylate 2.5%) 0.1 % 첨가구, $Lactacid^(R)$ 0.5% 첨가구, 그리고 $Immunocin^(R)$ 0.1% 첨가구등 4첨가구를 두었다. 생산성에서 essential oil 복합제 $Immunocin^(R)$ 0.1% 첨가구가 $BMD^(R)$ 25ppm 첨가한 항생제구와 동일한 수준의 생산성 개선 효과를 보였는데 (P<0.05) 특히 사료 요구율이 유의하게 향상되었다(P<0.05). 그러나 유기산 복합제인 $Lactacid^(R)$ 0.5% 첨가구는 항생제구보다 생산성 개선 효과가 없었다. IgG 농도는 $Immunocin^(R)$구가 첨가구들 중에서 가장 높았으며(P<0.05) 혈액의 leukocytes 와 stress index는 $Immunocin^(R)$ 첨가구가 낮은 경향을 나타냈다. 장내 미생물의 Cl. perfringens 수는 첨가구간에는 유의차가 없었으며 E. coli 수는 항생제구와 $Immunocin^(R)$ 구가 가장 적었다(P<0.05). 영양소 이용 율에 있어서 조단백질은 $Lactacid^(R)$ 구와 $Immunocin^(R)$ 구 높았다(P< 0.05). NFE 이용율은 생산성 개선 효과는 없었으나 $Lactacid^(R)$ 구가 가장 높았다(P<0.05). 결론적으로 essential oil 제제인 $Immunocin^(R)$을 육계 사료의 첨가제로 사용시 항생제와 같은 수준의 생산성 개선 효과를 보였으나 유기산제제인 $Lactacid^(R)$는 대조구와 차이가 없었다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권2호
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• pp.246-252
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• 2010

In order to improve the availability of phytase and probiotics together, a phytase gene from Aspergillus ficuum has been expressed in Lactobacillus. In this study, the transformed Lactobacillus with phytase gene was fed to pigs to determine its effect on pig production, feed conversion and gut microbes. Forty eight, 60-day-old, castrated pigs (Duroc${\times}$Landrace${\times}$Pietrain) were assigned to 6 groups, 8 pigs for each group. Group 1 was the control, group 2 was added with chlortetracycline (500 mg/kg), group 3 was added with the transformed Lactobacillus (500 mg/kg) with 20% (w/w) of calcium monohydrogen phosphate (CMP, $CaHPO_{4}$) removed, group 4 was added with the natural Lactobacillus (500 mg/kg) with 20% (w/w) of CMP removed, group 5 was added with the transformed Lactobacillus (500 mg/kg) with 40% (w/w) of CMP removed, group 6 was added with phytase (500 mg/kg) with 40% (w/w) of CMP removed. The results showed: i) the average daily gain (ADG) was improved in groups 2, 3 and 4 (p<0.05); ii) the diarrhea rates in the groups added with Lactobacillus were lower than in the other groups (p<0.05), in which the transformed Lactobacillus had more effect on reducing digestive disease; iii) the transformed Lactobacillus was most effective in improving the digestibilities of crude protein (CP), calcium (Ca), phosphorus (P), compared with the other groups (p<0.05); iv) Lactobacillus could increase lactic acid bacterium number and ammonia concentrations, and decrease pH values and E. coli number in pig feces (p<0.05); v) the phytase activity in the feces of pigs fed with the transformed Lactobacillus was 133.32 U/g, which was higher than in group 4 (9.58 U/g, p<0.05), and was almost the same as group 6 (135.94 U/g); vi) the transformed Lactobacillus could increase serum concentrations of IgA, triglyceride, and glutamic oxaloacetic transaminase activity (p<0.05), and had no significant effect on other serum indexes (p>0.05).

• 한국동물위생학회지
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• 제43권1호
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• pp.23-29
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• 2020

Hepatitis E Virus (HEV) infection is a worldwide disease and the primary cause of acute viral hepatitis in the world. It can be isolated from many different species including pigs. HEV is a zoonotic pathogen and foodborne disease. The main animal reservoir is domestic pigs. It is usually asymptomatic in pig but it is a public health concern, causing acute hepatitis in humans of varying severity. This study focused on the presence of HEV in pig and pork product. One hundred feces and one hundred fifty serum samples were randomly collected from pigs in slaughterhouses in Gwangju from November in 2018 to February in 2020. In addtion, seventy-five pork products were collected from markets in Gwangju. Feces and pork product samples were examined for the presence of HEV RNA using an reverse-transcription realtime PCR (RT-qPCR) assay. Serum samples were tested for the presence of HEV-specific IgG antibodies using Enzyme-linked immunosorbent assay (ELISA). HEV antigen and antibody positive rates were 3.0% (3/100) and 19.3% (29/150), respectively, in Gwangju and nearby areas such as Jeonnam and Jeonbuk. However, HEV antigen was not detected from any of pork product in this study. In conclusion, the prevalence of HEV should be continuously monitored because HEV was sporadically detected in Gwangju and nearby areas.

• 동의생리병리학회지
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• 제32권4호
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• pp.255-260
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• 2018

Chung-Sang (CS) is an experimental herbal remedy that is formulated to treat respiratory diseases implicated by inflammation. The herbs comprised of CS are frequently prescribed for treating various inflammatory symptoms: Menthae haplocalycis Herba, Magnoliae Flosis, Xanthii Fructus, Herba Asari, and Caryphylli Flos. Here, we prepared the extract of CS with boiling water (wCS) or with 50 % ethanol (eCS) and examined whether the two different extracts of CS exhibit a toxicity to cultured cells and mice. RAW 264.7 cells were treated with wCS or eCS, and the cytotoxicity of these extracts to RAW 264.7 cells was determined by an MTT assay. Although the production of intracellular reactive oxygen species that are detrimental to the cells was not increased by the extracts, the cytotoxicity to the cells was evident from 10 mg/ml of wCS and 100 mg/ml of eCS, suggesting that eCS is less cytotoxic. When mice (n = 10/group) received a single intratracheal wCS or eCS daily for 14 days, wCS yielded 40 % mortality, whereas eCS showed none. Both wCS and eCS did not significantly affect the weight of the body and of vital organs, except the lung. Biochemical analyses of mice blood indicated no damage to liver or kidney. However, unlike eCS, wCS significantly increased the level of IgE in serum. Collectively, our results show that eCS was less toxic than wCS, suggesting that CS prepared with 50 % ethanol is preferential over the conventional way of preparing CS.

• 한국가금학회지
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• 제35권1호
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• pp.71-78
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• 2008

본 실험은 Herbs와 Plant extracts(PE)가 산란계의 생산성과 소장내 미생물 균총 및 면역 체계에 미치는 영향을 조사하기 위해 실시하였다. 사양 시험은 67주령의 산란계(Hy-line Brown) 1,440수를 선별하여 A형 2단 케이지에 대조구 포함 총 9처리구로 구성하여 처리당 8반복 반복당 20수씩 randomized block design으로 배치하였다. 시험 기간 동안 물과 사료는 자유 섭취하게 하였으며, 일반적인 점등 관리(자연 일조 + 조명; 16 hr)를 실시하였다. 시험구는 대조구, 항생제구; Avilamycin 6 ppm, $Herb-Mix^{(R)}$ 0.2%구, PE-BIOSTRONG $510^{(R)}$ 0.02% 구, $PE-APEX^{(R)}$ 0.02%구, $PE-Digestarom^{(R)}$ 0.02%구, $PE-Phello-zyme^{(R)}$ 0.1%구, $PE-Galicin^{(R)}$을 0.05%구, PE-CRINA $Poultry^{(R)}$ 0.05%구 등 총 9처리구였다. 일계 산란율은 처리구간에 유의적 차이는 없었지만, 첨가구들이 대조구보다 높은 경향을 나타냈다. 사료 섭취량과 사료 요구율은 처리구간에 유의한 차이를 보였다. 사료 섭취량은 첨가구들이 대조구보다 높았다. 사료 요구율은 T8(PE-Galicin)구가 모든 첨가구중에 가장 높았다. 난각과 난황 색깔 지수는 처리구간에 유의한 차이가 있었다. 난각 색깔 지수는 첨가제간에 유의한 차이가 있었으며, 난황 색깔 지수는 T9(PE-CRINA Poultry)가 대조구보다 유의하게 높았다. Haugh unit은 처리구간에 유의적인 차이는 나타나지 않았다. Leukocytes와 erythrocytes 관련 모수들은 처리간에 유의한 차이가 있었다. WBC 수준과 stress index는 일반적으로 첨가구들이 대조구보다 높았다. RBC 수준은 Herb-Mix와 PE구들이 대조구보다 낮은 경향을 보여주었다. 혈청 IgG 농도는 처리구간에 유의적인 차이는 없었지만 모든 첨가구들이 대조구보다 높았다. 첨가구들에서 Lactobacillius는 증가하고 Cl. perfrigens는 감소하였다. E. coli는 첨가구들이 대조구보다 유의하게 낮았다. 결론적으로 herbs와 plant extract는 산란계에서 산란율을 증가시키는 경향이 있었고 혈액 성상 및 장내 미생물 균총에 유의한 영향을 미쳤다.

• 한국식품영양과학회지
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• 제39권11호
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• pp.1611-1618
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• 2010

본 연구는 in vitro 실험결과 아토피 피부염의 억제 효능이 기대되는 비파엽 추출물(BI군), 삼백초 추출물(SA군), 비파엽 추출물과 삼백초 추출 혼합물(FB군) 그리고 삼백초와 비파엽 추출 혼합 발효물(FA군)을 경구 투여하여 DNCB 처리한 아토피 실험동물을 대상으로 아토피 질환 개선에 대한 유효성을 평가하였다. 혈청 중의 IgE 수치를 측정한 결과, DNCB군에서 $227.21{\pm}60.04$ ng/mL를 나타내었으며, FA군에서는 $154.78{\pm}78.99$ ng/mL로 IgE 생성량이 감소되었다. 혈청내의 히스타민 농도는 DNCB군에서 $1.90{\pm}0.04$ ng/mL 였으며, FA, FB, BI, SA군에서 각각 $1.47{\pm}0.20$ ng/mL, $1.41{\pm}0.07$ ng/mL, $1.61{\pm}0.05$ ng/mL, $1.08{\pm}0.07$ ng/mL로 DNCB군과 비교 시 유의적인 감소를 나타내었다. 세라마이드 총 함량을 측정한 결과, FA군에서 $1,350{\pm}357\;{\mu}g/g$으로 DNCB와 비교 시 세라마이드 함량이 유의적으로 증가되었다. IFN-$\gamma$의 경우, DNCB군에서 $2.5{\pm}0.5$ ng/mL로 분석되 었으며, FA, FB, BI 및 SA군에서 각각 $7.1{\pm}1.3$ ng/mL, $7.7{\pm}0.2$ ng/mL, $11.5{\pm}0.6$ ng/mL 및 $6.9{\pm}0.2$ ng/mL로 DNCB 군과 비교 시 모두 유의적인 증가를 나타내었다. 특히 비파엽 및 삼백초 추출물 혼합 발효물은 등 부위 피부의 육안 증상을 관찰한 결과, 가피형성과 발적 감소가 가장 우수하여 아토피 피부염 질환 개선에 도움을 줄 수 있을 것으로 기대된다.

• 한국미생물·생명공학회지
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• 제36권4호
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• pp.343-352
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• 2008

프랑킨센스는 감람나무 종에서 얻어지는 gum resin으로, 성분은 5-9% 방향정 에센셜 오일, 알코올-용해성인 65-85% resin과 수용성인 gum 잔여물로 구성된 복합물이다. 프랑킨센스의 알코올 용해성인 resin 성분들의 항염증 작용은 잘 알려져 있으나, 방향성 에센셜 오일 성분이 알러지성 천식에 작용을 하는지의 여부는 보고되지 않았다. 실험은 프랑킨센스 에센설 오일(BSEO)이 ovalbumin(OVA)으로 유발된 알러지성 천식 생쥐 모델에 미치는 영향을 조사하기 위해 수행하였다. BALB/c 생쥐는 OVA로 복강감작 후 OVA 기도 투여로 면역반응을 유발시켰다. 실험그룹은 0.3% BSEO를 8주간 흡입시켰다. OVA로 감작, 유발시킨 BALB/c 생쥐에서 기도내 호산구 침윤증가, 점액분비 증가와 기도과민성이 나타났다. 이에 비하여, BSEO 처치군에서 BALF내 호산구수, 술잔세포의 과증식, 기도과민성이 감소되었다. BALF내 사이토카인 분석 결과, BSEO는 Th1 사이토카인인 IFN-$\gamma$를 증가시켰으며 Th2 사이토카인인 IL-4, IL-5와 IL-13을 감소시켰다. 또한, OVA-specific IgE와 eoxtain 분비를 억제시켰다. BSEO 흡입 군에서 종격동 림프절의 $CD4^+$, $CD3^+/CCR3^+$, 및 $B220^+/CD23^+$ 세포들 또한 감소되었다. 이상의 결과에서 BSEO는 Th1/Th2 관여 면역조절인자로 판단되며, BSEO 흡입으로 간단하고 경제적인 방법으로 알러지성 기도 염증 치료가 가능할 것으로 사료되었다.