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SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

Anti-diabetic effect and mechanism of Korean red ginseng extract in C57BL/KsJ db/db mice

  • Yuan, Hai-Dan;Shin, Eun-Jung;Chung, Sung-Hyun
    • Proceedings of the Ginseng society Conference
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    • 2007.12a
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    • pp.57-58
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    • 2007
  • Purpose: Ginseng is a well-known medical plant used in traditional Oriental medicine. Korean red ginseng (KRG) has been known to have potent biological activities such as radical scavenging, vasodilating, anti-tumor and anti-diabetic activities. However, the mechanism of the beneficial effects of KRG on diabetes is yet to be elucidated. The present study was designed to investigate the anti-diabetic effect and mechanism of KRG extract in C57BL/KsJ db/db mice. Methods: The db/db mice were randomly divided into six groups: diabetic control group (DC), red ginseng extract low dose group (RGL, 100 mg/kg), red ginseng extract high dose group (RGH, 200 mg/kg), metformin group (MET, 300 mg/kg), glipizide group (GPZ, 15 mg/kg) and pioglitazone group (PIO, 30 mg/kg), and treated with drugs once per day for 10 weeks. During the experiment, body weight and blood glucose levels were measured once every week. At the end of treatment, we measured Hemoglobin A1c (HbA1c), blood glucose, insulin, triglyceride (TG), adiponectin, leptin, non-esterified fatty acid (NEFA). Morphological analyses of liver, pancreas and white adipose tissue were done by histological observation through hematoxylin-eosin staining. Pancreatic islet insulin and glucagon levels were detected by double-immunofluorescence staining. To elucidate an action of mechanism of KRG, DNA microarray analyses were performed, and western blot and RT-PCR were conducted for validation. Results: Compared to the DC group mice, body weight gain of PIO treated group mice showed 15.2% increase, but the other group mice did not showed significant differences. Compared to the DC group, fasting blood glucose levels were decreased by 19.8% in RGL, 18.3% in RGH, 67.7% in MET, 52.3% in GPZ, 56.9% in PIO-treated group. With decreased plasma glucose levels, the insulin resistance index of the RGL-treated group was reduced by 27.7% compared to the DC group. Insulin resistance values for positive drugs were all markedly decreased by 80.8%, 41.1% and 68.9%, compared to that of DC group. HbA1c levels in RGL, RGH, MET, GPZ and PIO-treated groups were also decreased by 11.0%, 6.4%, 18.9%, 16.1% and 27.9% compared to that of DC group, and these figure revealed a similar trend shown in plasma glucose levels. Plasma TG and NEFA levels were decreased by 18.8% and 16.8%, respectively, and plasma adiponectin and leptin levels were increased by 20.6% and 12.1%, respectively, in the RGL-treated group compared to those in DC group. Histological analysis of the liver of mice treated with KRG revealed a significantly decreased number of lipid droplets compared to the DC group. The control mice exhibited definitive loss and degeneration of islet, whereas mice treated with KRG preserved islet architecture. Compared to the DC group mice, KRG resulted in significant reduction of adipocytes. From the pancreatic islet double-immunofluorescence staining, we observed KRG has increased insulin production, but decreased glucagon production. KRG treatment resulted in stimulation of AMP-activated protein kinase (AMPK) phosphorylation in the db/db mice liver. To elucidate mechanism of action of KRG extract, microarray analysis was conducted in the liver tissue of mice treated with KRG extract, and results suggest that red ginseng affects on hepatic expression of genes responsible for glycolysis, gluconeogenesis and fatty acid oxidation. In summary, multiple administration of KRG showed the hypoglycemic activity and improved glucose tolerance. In addition, KRG increased glucose utilization and improved insulin sensitivity through inhibition of lipogenesis and activation of fatty acid $\beta$-oxidation in the liver tissue. In view of our present data, we may suggest that KRG could provide a solid basis for the development of new anti-diabetic drug.

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A Study on the Gene Expression of Adipogenic Regulators by an Herbal Composition (생약복합물에 의한 지방세포형성 조절자의 유전자 발현 연구)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.5
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    • pp.729-735
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    • 2010
  • In our previous study, it was reported that an herbal mixture, SH21B, inhibits fat accumulation and adipogenesis both in vitro and in vivo models of obesity. SH21B is a mixture composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd, and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). The aim of this study was to investigate the detailed molecular mechanisms of the effects of SH21B on various regulators of the adipogenesis pathway. During the adipogenesis of 3T3-L1 cells, SH21B significantly decreased the expression levels of central transcription factors of adipogenesis, such as peroxisome proliferator-activated receptor (PPAR)$\gamma$ and CCAAT/enhancer binding protein (C/EBP)$\alpha$. To elucidate the detailed molecular mechanism of the anti-adipogenic effects of SH21B, we examined the expression levels of the various pro-adipogenic or anti-adipogenic regulators of adipogenesis upstream of $PPAR{\gamma}$ and C/$EBP{\alpha}$. The mRNA levels of Krox20 and Kruppel-like factor (KLF) 15, which are pro-adipogenic regulators, were significantly down-regulated by SH21B treatment, whereas the mRNA levels of C/$EBP{\gamma}$ and KLF5 were not changed. KLF2 and C/EBP homologous protein (CHOP), which are anti-adipogenic regulators, were significantly up-regulated by SH21B treatment. These results suggest that the molecular mechanism of the anti-adipogenic effect of SH21B involves both the down-regulations of pro-adipogenic regulators, such as Krox20 and KLF15, and the up-regulations of anti-adipogenic regulators, such as KLF2 and CHOP, which results in the suppression of central transcription factors of adipogenesis including $PPAR{\gamma}$ and C/$EBP{\alpha}$.

Failure Behavior and Separation Criterion for Strengthened Concrete Members with Steel Plates (강판과 콘크리트 접착계면의 파괴거동 및 박리특성)

  • 오병환;조재열;차수원
    • Journal of the Korea Concrete Institute
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    • v.14 no.1
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    • pp.126-135
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    • 2002
  • Plate bonding technique has been widely used in strengthening of existing concrete structures, although it has often a serious problem of premature falure such as interface separation and rip-off. However, this premature failure problem has not been well explored yet especially in view of local failure mechanism around the interface of plate ends. The purpose of the present study is, therefore, to identify the local failure of strengthened plates and to derive a separation criterion at the interface of plates. To this end, a comprehensive experimental program has been set up. The double lap pull-out tests considering pure shear force and half beam tests considering combined flexure-shear force were performed. The main experimental parameters include plate thickness, adhesive thickness, and plate end arrangement. The strains along the longitudinal direction of steel plates have been measured and the shear stress were calculated from those measures strains. The effects of plate thickness, bonded length, and plate end treatment have been also clarified from the present test results. Nonlinear finite element analysis has been performed and compared with test results. The Interface properties are also modeled to present the separation failure behavior of strengthened members. The cracking patterns as well as maximum failure loads agree well with test data. The relation between maximum shear and normal stresses at the interface has been derived to propose a separation failure criterion of strengthened members. The present study allows more realistic analysis and design of externally strengthened flexural member with steel plates.

Introduction of Integrated Coastal Management Program and Sustainable Development of Fishing Villages in Cheonsu Bay Region (연안통합관리계획의 도입과 천수만 어촌의 지속가능발전)

  • 김부성
    • Journal of the Korean Geographical Society
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    • v.38 no.2
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    • pp.184-205
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    • 2003
  • Sustainable Development(SD) is an important concept for the future of the coastal area, and for development of fishing villages. Since 1992 UN Conference on Environment and Development in Rio de Janeiro many governments and local authorities throughout the world have been engaged in preparing and implementing $\ulcorner$Agenda 21$\lrcorner$. Many projects which previously would have been identified as environmental protection are now presented under the banner of sustainable development. Integrated Coastal Management (ICM) is an extension of sustainable development. ICM was presented as a framework for resolution of coastal use conflicts. The aim of the present paper is to assess sustainable development potential of fishing villages in Cheonsu Bay Region according to implementation of ICM. Cheonsu Bay Region was known as one of the productive fishing grounds and Cheonsu Bay Region preserved unique characteristics of traditional fishing villages. But this region is now experiencing many changes through the massive reclamation projects like Seosan A B Project. After a brief overview of concepts and history of SD and ICM, the reclamation process and its impacts on both fishery and fishing communities in Cheonsu Bay Region are discussed. According to their changing environmental and socio-economic characteristics after the reclamation, ca 35 representative coastal villages in this region can be classified into 5 types. Many coastal villages shows diversity in their economic activities, as tourism and recreation function becomes more and more important in this region. In present-day Cheonsu Bay Region, it is possible to differentiate fishing village cooperatives(FVO) with high potential of sustainable fishery development, FVOs with medium potential, FVOs with low potential on the basis of 14 selected indicators.

Incidence of Escherichia coli and Its Susceptibility to Antimicrobials in Childhood Urinary Tract Infection (소아 요로감염에서 Escherichia coli의 빈도와 항생제 감수성에 대한 연구)

  • Chi Hye-Mi;Kwahk Jae-Hyok;Lee Jun-Ho;Park Hye-Won
    • Childhood Kidney Diseases
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    • v.10 no.1
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    • pp.18-26
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    • 2006
  • Purpose : Empirical antimicrobial treatment is indicated before bacteriological results are available for young children with febrile UTI to minimize renal scarring. To ensure appropriate therapy, knowledge of the prevalence of causative organisms and their susceptibility patterns to antimicrobials is mandatory. We performed a retrospective analysis investigating the local prevalence and resistance patterns of uropathogens, primarily E. coli, isolated from community-acquired UTIs. Methods : A total of 103 positive urine cultures from children with febrile UTI collected at Bundang CHA General Hospital from February 2004 to February 2005 were analyzed. Inclusion criteria were fever higher than $37.5^{\circ}C$, significant bacteriuria with single strain growth of at least 10s colony forming units/mL urine, and leukocyturia >5/HPF. Results : E. coli(89.3%) was the leading uropathogen followed by Enterococcus spp.(3.9%) Klebsiella spp.(2.9%), Citrobctcter spp.(1.9%) and Enterobacter spp.(1.9%). E. coli strains revealed a low proportion of antimicrobial susceptibility to ampicillin(AMP; 27.2%) ampicillinsulbactam(AMS; 34.8%) and trimethoprim-sulfamethoxazole(SXT; 65.2%). Susceptibility patterns to cephalosporins were as follows; cefazolin(1st generation; 91.3%), cefoxitin(2nd; 100%), ceftriaxone(3rd; 97.8%) and cefepime(4th; 97.8%). Three E. coli isolates produced ex tended - spectrum beta-lactamase(ESBL). Conclusion : Empirical treatment with AMP, AMS and SXT, which are commonly used in pediatric clinics, is not recommended for childhood UTI due to high incidence of resistance. The high level of susceptibility to cephalosporins makes these drugs reasonable alternatives. However the emergence of ESBL-producers, even though they are quite few, may have an impact on cephalosporin treatment in the future. (J Korean Soc Pediatr Nephrol 2006;10:18-26)

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Antioxidant and Anti-aging Effects of Extracts from Leaves of the Quercusaliena Blume on Human Dermal Fibroblast (피부 섬유아세포에서 갈참나무 잎 추출물의 항산화 및 항노화 효능)

  • Choi, Sun-Il;Lee, Jong Seok;Lee, Sarah;Yeo, Joohong;Jung, Tae-Dong;Cho, Bong-Yeon;Choi, Seung-Hyun;Sim, Wan-Sup;Han, Xionggao;Lee, Jin-Ha;Kim, Jong Dai;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.140-145
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    • 2018
  • The skin of the human body occupies the largest surface area of the body and acts as a protection for the person's internal organs. As such, the skin is a major target of oxidative stressors, and these oxidative stressors are known to contribute to skin aging over the course of time. For the most part, an antioxidant is an effective approach to utilize to prevent symptoms related to the reactive oxygen species (ROS)-induced aging of the skin. Therefore, we investigated the antioxidant and anti-aging activity of the leaves of the Quercusaliena Blume extract (QBE). In our study, we confirmed that the cell viability tested with XTT {2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide innersalt} assay was not affected up to a concentration of $100{\mu}g/mL$. In addition, the cell viability of HDF cells induced by hydrogen peroxide was recovered from 81% to 104% after treatment with QBE, which showed the greater protective effect than that of ascorbic acid. Treatments of QBE dose-dependently inhibited reactive oxygen species (ROS) production in HDF cells induced by hydrogen peroxide, which correlated with their protective effects on cell viability. Since QBE treatment exhibited the suppression effect of skin aging by decreasing the ROS production, QBE could be used as a not only natural anti-aging but also antioxidant resource.

Quality characteristics of spread jam using low-salt Lentinula edodes soybean paste (저염표고된장을 활용한 스프레드잼의 품질특성)

  • Ha, Neul-I;Jeong, Hee-Gyeong;Jin, Seong-Woo;Kim, Kyung-Je;Koh, Young-Woo;Im, Seung-Bin;Jeong, Sang-Wook;Yun, Kyeong-Won;Kim, Ki-Man;Seo, Kyoung-Sun
    • Journal of Mushroom
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    • v.19 no.3
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    • pp.200-209
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    • 2021
  • In this study, a low-salt Lentinula edodes soybean paste (LSLESP) with improved palatability and storability relative to traditional Korean soybean paste was developed by utilizing low quality log cultivation of L. edodes. We also conducted quality characteristic analyses of spread jam made from low-salt L. edodes soybean paste (SJLLESP). The lowest salinity content and the highest L (brightness) value was found in LSLESP. According to proximate compositions analysis, crude protein, crude fat, and ash content were higher in commercial soybean paste than in LSLESP. Glucose, fructose, and maltose were the only free sugars detected in commercial soybean paste, whereas arabinose and fucose were additionally detected in LSLESP. Only two types of organic acid, were detected in common in commercially available soybean paste and LSLESP. Four types of organic acid, were detected in SJLLESP due to its added ingredients. The major free amino acids in commercially available soybean paste and LSLESP are histidine, glutamic acid, and arginine. Total amino acid, content was higher in LSLESP (54.81 mg%) than in commercial soybean paste (49.26 mg%). Total free amino acid, content in SJLLESP was 43.01 mg%. Ergosterol and β-glucan contents were highest in SJLLESP. The elevated ergosterol and β-glucan content in low-salt LSLESP relative to commercial soybean paste was significant. In conclusion, LSLESP and SJLLESP contain useful components from L. edodes, and offer the advantage of low salinity. LSLESP and SJLLESP could thus contribute to the development of health foods using L. edodes.

Floristic Characteristics of Vascular Plants in the Maryong-san(Mt. 406.7m) and Daeso-san(Mt. 286m) Area (마룡산(406.7m, 울진군)과 대소산(286m, 영덕군) 일원의 관속식물)

  • Kim, Young-Chul;Chae, Hyun-Hee;Park, You-Cheol;Lee, Seon-Mi;Nam, Gi-Heum
    • Korean Journal of Environment and Ecology
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    • v.35 no.5
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    • pp.438-479
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    • 2021
  • Basic information such as flora and their distribution are the key factors to consider for preserving plant diversity on a regional level. The number of plant species identified through a survey is closely related to the survey area and the number of habitats distributed within the survey area. In this study, we surveyed the flora of Mt. Maryong-san (PH) and Mt. Daeso-san (YH) located in Uljin-gun, Gyeongsangbuk-do, six and nine times, respectively. We planned to include a variety of habitats in the survey routes. As a result, we identified a total of 594 taxa composing of 112 families, 349 genera, 548 species, 10 subspecies, 34 varieties, and 2 formulas in the Mt. Maryong-san (PH) area and a total of 434 taxa composing of 89 families, 270 genera, 400 species, 9 subspecies, and 25 varieties in the Mt. Daeso-san (YH) area. Of the plants observed in the 21 habitats, the plants distributed in the transition zone around the valley and the dry grassland accounted for the highest proportion. The traditional graveyards located on the coast also served as the major habitat for the distribution of some plant species. In the two graveyards, 19 taxa of endemic plant species were surveyed. Moreover, the identified floristic target species included 1 taxon for grade V, 11 taxa for grade IV, and 24 taxa for grade III. A total of 77 taxa of naturalized plants were identified, and four of them were invasive alien plants as designated by the Ministry of Environment. The differences in the plants distributed in the area of Mt. Maryong-san (PH) and Mt. Daeso-san (YH) were mainly due to the difference in the size of each habitat distributed within the survey area. Moreover, it was judged that the entire vicinity of Mt. Daeso-san (YH) was not connected to the surrounding mountains and was an isolated area surrounded by farmland and residential areas.

Immune-enhancing and Anti-inflammatory Effects of HK Shiitake Mushroom Mycelium (HKSMM) using Balb/c Mice (Balb/c 마우스를 이용한 HK 표고버섯 균사체(HKSMM)의 면역증강 및 항염효과)

  • Kim, Hun Hwan;Ha, Sang Eun;Park, Min Young;Jeong, Se Hyo;Bhagwan, Bhosale Pritam;Abuyaseer, Abusaliya;Kim, Jeong Ok;Ha, Yeong Lae;Kim, Gon Sup
    • Journal of Life Science
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    • v.32 no.6
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    • pp.447-454
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    • 2022
  • In this study, we confirmed the effect of HK shiitake mushroom mycelium (HKSMM) on immune enhancement in Balb/c mice. Experimental animals were divided into five groups: negative control (NC), positive control (PC; 1,000 mg/100 g; AHCC), T1 (500 mg/100 g; HKSMM), T2 (1,000 mg/100 g; HKSMM), and T3 (2,000 mg/100 g; HKSMM), and dissection was performed at four and six weeks. COX-2 and iNOS concentrations were significantly lower in the six-week experimental group than in the control group, and the NO results were also similar. Results of the confirmation of the factors related to the NF-κB (p-p65 and p-IκBα) and MAPK (pERK, pJNK, and p38) signaling pathways revealed that the HKSMM-fed experimental group significantly decreased compared with the control group. A comparative analysis of the number and size of white pulp in the spleen tissue showed that those of the experimental group were significantly higher than those of the control group in a concentration-dependent manner. These results suggest that HKSMM has both immune-enhancing and anti-inflammatory effects in Balb/c mice, indicating that it can be used as a health functional food ingredient.