• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.72-78
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• 2001

It is possible ot prepare protoplasts of the zygomycete fungus, Phycomyces blakesleeanus, by digesting the cell wall of spore germlings with commercially available chitinase and chitosanase. However, the cells without any cell walls immediately form large aggregates, and thus, it is difficult to isolate the individually separated protoplasts. Inherent problem with the formation of aggregates in preparing protoplasts could be solved by the use of bovine serum albumin (BSA). As a result, we were able to prepare a large number of single protoplsts quickly and easily. We took time-lapse photomicrographs of the formation of protoplasts, and found that there were certain regions of the cell wall of spore germlings that were sensitive to chitinase and chitosanase, although the cell wall of the original spores is known to be insensitive to these enzymes. There are two kinds of cell walls on a spore germling; one with a bound wheat germ agglutinin (WGA), and the other a bound concanavalin A (ConA). Furthermore, only cells with walls which had bound WGA were able to regenerate, while those with walls with bound ConA were not able to regenerate.

• 생명과학회지
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• 제32권9호
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• pp.678-689
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• 2022

HK shiitake mushroom mycelium (HKSMM: 14% β-glucan)은 식품의약품안전처의 개별인정형 간 건강기능식품 원료이다. HKSMM의 50% ethanol 추출물 (HKSMM50)에 대한 면역증강 효과를 Concanavalin A (ConA)로 활성화한 human T lymphocyte Jurkat cells에서 연구하였다. Active hexose correlated compound (AHCC)는 positive control로 사용하였다. ConA로 활성화한 Jurkat cells에 HKSMM50 (0, 25, 50, 100 ㎍ g/ml) 및 AHCC (100 ㎍ g/ml)를 처리하고 3시간 또는 6시간 배양하였다. Jurkat cells의 cytosol과 nucleus에 함유된 transcription factor인 nuclear factor of activated T cells (NFAT) 함량은 Western blotting으로 측정하였다. Interleukin-2 (IL-2)와 interferon-gamma (IFN-γ) 함량 및 cyclooxygenase-2 (COX-2) 활성은 enzyme-linked immunosorbent assay (ELISA) kit로 분석하였다. HKSMM50은 cytosolic NFAT protein 함량은 낮추었고, nuclear NFAT protein함량은 증가시켰다. IL-2와 IFN-γ 함량은 증가되었고, COX-2 활성과 apoptosis는 억제되었다. AHCC 효과는 HKSSM50의 효과와 유사하였다. 이와 같은 결과는 HKSMM50가 ConA로 활성화된 Jurkat cells에서 NFAT protein을 활성화시켜, IL-2와 IFN-γ 함량을 증가시켰음을 의미한다. 또한 HKSMM50은 COX-2 활성과 apoptosis를 억제하였다. 따라서 이와 같은 결과는 HKSMM이 면역증진을 위한 건강기능식품 원료로 사용할 수 있음을 의미한다.

• Journal of Evidence-Based Herbal Medicine
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• 제2권1호
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• pp.19-24
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• 2009

GD68 is newly developed herb complex prescription. The constituent herbs of GD68 were Massa Medicata Fermentata, Atractylodis Rhizoma Alba, Poria, Zingiberis Siccatum Rhizoma, Crataegi Fructus, Saccarum Granorum, Agastachis Herba, Taraxaci Herba, Perillae Herba, Scutellariae Radix, Astragali Radix, Ginseng Radix, Houttuyniae Herba and Halloysitum Rubrum. The aim of this study was to examine feed value of GD68 in duck. The weight gain of ducks fed with supplemental GD68 high compared to those of the control. The feed intake and mortality of ducks fed with supplemental GD68 low compared to those of the control. The moisture, crude lipid and calorie content of the ducks fed GD68 were decreased, but the crude protein content of the ducks fed GD68 was increased. And we investigated the effect of GD68 on the production of cytokines in human T-cell line, MOLT-4 cells. GD68 plus concanavalin A (Con A) increased the interferon-$\gamma$ and interleukin-2 production compared with Con A alone. These results indicate that the supplemental GD68 may improve the production, meat quality and immunity of ducks.

• 한국응용과학기술학회지
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• 제40권5호
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• pp.955-964
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• 2023

Bee pollen is a valuable apitherapeutic product and has been known to have diverse biological activities, including antimicrobial, anti-inflammatory, and even anticancer activity. However, its effect on the immune system is not well studied and is rather controversial. This study intended to elucidate the biological activity of bee pollen on immunity. For this purpose, we used lyophilized bee pollen after wet grinding, which shows increased extraction of bioactive components and enhanced biological activity. First, lyophilized bee pollen after wet grinding significantly increased the proliferation of splenocytes isolated from normal mice. On the other hand, lyophilized bee pollen after wet grinding dose-dependently reversed splenocyte proliferation by concanavalin A or lipopolysaccharide. To clarify the activity of bee pollen on immunity lyophilized bee pollen after wet grinding was administered daily to mice for five weeks and isolated splenocytes. In this study, there was no significant difference in the population of immune cells and the size of spleen between bee pollen- and sterile water-treated groups. However, proliferation of splenocyte isolated from bee pollen-administered animals was boosted by both concanavalin A and lipopolysaccharide. Finally, kaempferol, a well-known flavonoid from bee pollen, dose-dependently increased splenocyte proliferation by both Con A and LPS. On the other hand, naringenin, another flavonoid in the bee pollen, dose-dependently inhibited the proliferation of splenocytes by Con A and LPS. Together, these data indicate that bee pollen may be able to prime the immunity to boost immune reaction after inflammation.

• 대한한의학회지
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• 제22권1호
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• pp.46-52
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• 2001

Objectives: This experimental study was carried out to evaluate the effects of PSM(polysaccharide of mushroom) on the immune activity. Methods: The following were performed; Immunotoxicity testing for immunopathology, IgO production & LPS mitogen response for humoral immunity, DTH, ConA mitogen response for cell-mediated immunity, and macrophage adherence & phagocytosis for nonspecific immunity in vitro or in vivo. Results: PSM showed a protective effect on cyclophosphamide-induced leukopenia, increased IgG production and lymphoproliferative responses to LPS; inhanced DTH and lymphoproliferative response Con A; and activated macrophage adherence and phagocytosis to SRBC. Conclusions: It is suggested that PSM can be used for cancer patients with immunosuppression and adapted to many other diseases.

• 농업과학연구
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• 제36권1호
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• pp.41-50
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• 2009

Fucoidan is a uniquely-structured sulfated fucose-rich polysaccharide derived from brown algae. Recently, the abalone glycosidase-digested fucoidan extract (fucoidan extract) derived from seaweed Cladosiphon novae-caledoniae Kylin (Mozuku) draws much attention because of its clinical anti-cancer effect in Japan. Here, we report the cancer cells-specific apoptosis inducing effects of the fucoidan extract. The fucoidan extract suppressed the growth of various anchorage-dependent and -independent cancer cells. The fucoidan extract contained low molecular weight components, which induced apoptosis of human leukemic HL 60 cells but not of human lymphocytes. It was shown that the fucoidan extract lead caspase 3/7 activation and loss of mitochondrial membrane potential in HL 60 cells. Another function of the fucoidan extract was also observed. It has been known that sugar chain expression on the surface of cancer cell membrane changes dependent on their malignancy. The analysis on sugar chain expression profiling using FITC-labeled lectins revealed that the expression of concanavalin A (Con A) binding sugar chain was enhanced by the treatment of human lung adenocarcinoma A549, human uterine carcinoma HeLa and human fibrosarcoma HT1080 cells with the fucoidan extract. Con A-induced apoptosis of cancer cells was stimulated in a dose-and time-dependent manner by the treatment with the fucoidan extract but not of human normal fibroblast TIG-1 cells.

• Journal of Acupuncture Research
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• 제34권2호
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• pp.39-48
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• 2017

Objectives : In this study, we investigated the immunomodulatory and antioxidant effect of MOK, a pharmacopuncture medicine, in concanavalin A (Con A)-stimulated mouse splenocytes. Methods : Primary splenocytes were isolated from ICR mice. The splenocytes were treated with MOK extract (1.25, 2.5, 5, 10, and 20 mg/mL) for 30 min and then stimulated with Con A (200 ng/mL) for the indicated times. Cell viability of the splenocytes was measured using an MTT assay. The mRNA expression of Th1/Th2 cytokines ($IFN-{\gamma}$, IL-4, IL-10, and Foxp3) and antioxidant enzymes (HO-1 and MnSOD) was measured by RT-PCR. Results : Addition of MOK extract at 2.5, 5, and 10 mg/mL in Con A-stimulated splenocytes significantly decreased the production of $IFN-{\gamma}$ and significantly increased the expression of IL-4, IL-10, and Foxp3 mRNA. MOK extract also increased the mRNA expression of HO-1 and MnSOD in splenocytes. Conclusion : MOK extract modulated the Th1/Th2 immune response via the regulation of cytokine levels in splenocytes and exerted an antioxidant effect via the upregulation of antioxidant proteins.

• Toxicological Research
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• 제33권4호
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• pp.283-290
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• 2017

The host immune system is the first line of host defense, consisting mainly of innate and adaptive immunity. Immunity must be maintained, orchestrated, and harmonized, since overactivation of immune responses can lead to inflammation and autoimmune diseases, while immune deficiency can lead to infectious diseases. We investigated the regulation of innate and adaptive immune cell activation by Artemisia capillaris and its components (ursolic acid, hyperoside, scopoletin, and scopolin). Macrophage phagocytic activity was determined using fluorescently labeled Escherichia coli, as an indicator of innate immune activation. Concanavalin A (ConA)- and lipopolysaccharide (LPS)-induced splenocyte proliferation was analyzed as surrogate markers for cellular and humoral adaptive immunity, respectively. Neither A. capillaris water extract (WAC) nor ethanol extract (EAC) greatly inhibited macrophage phagocytic activity. In contrast, WAC suppressed ConA- and LPS-induced proliferation of primary mouse splenocytes in a dose-dependent manner. Similarly, EAC inhibited ConA- and LPS-induced splenocyte proliferation. Oral administration of WAC in mice decreased ConA- and LPS-induced splenocyte proliferation, while that of EAC suppressed LPS-induced splenocyte proliferation. Repeated administration of WAC in mice inhibited ConA- and LPS-induced splenocyte proliferation. Ursolic acid, scopoletin, and scopolin reduced ConA- and LPS-induced primary mouse splenocyte proliferation, while hyperoside did not show such activity. These results indicate that A. capillaris and its components, ursolic acid, scopoletin, and scopolin, suppress ConA- and LPS-induced adaptive immune cell activation. The results suggest that A. capillaris is useful as a regulator of adaptive immunity for diseases involving excessive immune response activation.

• 대한수의학회지
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• 제27권1호
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• pp.41-45
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• 1987

The present study was carried out to investigate the optimal conditions on blastogenesis of guinea pig lymphocytes. A microculture system in conjuction with a semiautomatic multiple sample harvester was used to study the in vitro optimal condition of guinea pig lymphocytes. Careful analysis of lymphocyte transformation to concanavalin A (Con A) and lipopolysaccharide (LPS) mitogen determined optimal conditions as: (a) 10% fetal bovine serum in RPMI-1640 medium (b) 48-hour culture period.

• 약학회지
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• 제49권1호
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• pp.25-29
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• 2005

This study was performed to investigate the potential of pear (Pyrus pyrifolia) as a immune-modulating functional food by assay of splenocytes proliferation and induction of cytokines (IFN-${\gamma}$, IL-4) in vitro. When mouse splenocytes were exposed to various concentration (0.16, 0.31, 0.63, 1.25, 2.50 mg/ml) of pear methanol extracts (P-M) without mitogens, splenocytes proliferation (SP) was significantly increased. Also, SP to mitogens, concanavalin A (Con A) and lipopolysaccharide (LPS) were significantly increased by P-M when compared with controls. When splenocytes were cultured with P-M in the presence of Con A, cytokine (IFN-${\gamma}$, IL-4) levels in culture supernatant were significantly enhanced in a dose-dependent manner except 2.5 mg/ml when compared with control group. Therefore, our study suggest that the pear has the potential of being an immune-modulating functional food.