• Title/Summary/Keyword: Immunohistochemistry

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Histological Comparative Study of Rabbit Maxillary Sinus Augmentation with Bio-Oss and β-TCP (Bio-Oss와 β-TCP를 이용한 토끼 상악동 거상술 후의 조직학적 비교 연구)

  • Moon, Yong-Suk
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1220-1232
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    • 2018
  • The purpose of this animal study was to evaluate, by histological analysis, bone regeneration in rabbit maxillary sinuses with an anorganic bovine graft (Bio-Oss) and a ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) grafting. Bilateral sinus augmentation procedures were performed in 12 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the Bio-Oss group, Bio-Oss was grafted and in the ${\beta}-TCP$ group, ${\beta}-TCP$ was grafted and covered by replaceable bony windows. The animals were sacrificed at 2, 4, and 8 weeks after the surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin-eosin, Masson trichrome, and tartrate-resistant acid phosphatase stains and also by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), type I collagen, and osteocalcin content. Histologically, new bone formation was found on the surface of Bio-Oss and ${\beta}-TCP$ particles from 2 weeks and continued to 8 weeks. Significant higher new bone formation was revealed in the ${\beta}-TCP$ group than in the Bio-Oss group at 8 weeks. The amount of graft materials was significantly decreased in the ${\beta}-TCP$ group and the number of osteoclasts was significantly increased in the ${\beta}-TCP$ group from 4 to 8 weeks. Immunoreactivity to PCNA was reduced at 8 weeks. The expression of type I collagen was significantly increased in the ${\beta}-TCP$ group at 2 weeks, but was significantly increased in the Bio-Oss group at 8 weeks. Immunoreactivity to osteocalcin was increased from 2 to 8 weeks. These histological results can help in the selection of graft materials for implants. Both Bio-Oss and ${\beta}-TCP$ are proven graft materials, however, these results indicate that ${\beta}-TCP$ showed better bone regeneration results in rabbit maxillary sinus augmentation.

The Effect of Antioxidant Vitamins on Aflatoxin $B_{1}-DNA$ Adduct the Formation in Aflatoxin $B_1$ Administered Mice Liver (항산화비타민이 Aflatoxin $B_1$ 투여 마우스의 간세포에서 Aflatoxin $B_{1}-DNA$ Adduct 형성에 미치는 영향)

  • Park, Seon-Ja;Kang, Sung-Jo;Park, Jung-Hyun;Oh, Sang-Suk;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.33 no.6
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    • pp.669-675
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    • 2001
  • The objective of this study was to examine the effects of antioxidant vitamins on the formation of $AFB_{1}-DNA$ adduct and $AFB_{1}-inducing$ cellular oxidative damage. Intraperitoneal(i.p.)injections of 10 mg/kg vitamin C(VC) and 63.8 mg/kg vitamin E(VE) were repeatedly administrated 4 times with 2 days interval to 6 week old male ICR mice. After one hour of vitamin treatments, 0.4 mg/kg $AFB_1$ was injected in $AFB_1$ plus vitamin treated groups by same way. On the other hands, $AFB_1$ treated group was only injected with $AFB_1$ by the same method described above without vitamins. According to quantitative analysis of the $AFB_1$ in mice serum by indirect competitive ELISA, 12.28 and 18.78 ng/mL were detected in $AFB_1-treated$ groups, but 7.60 and 4.85 ng/mL in $AFB_1$ plus VC and VE treated groups, respectively. 23.78, 25.48 ng/mL of $AFB_1-DNA$ adduct were detected in mice liver of $AFB_1$treated groups, while 5.26, 7.81 ng/mL in $AFB_1$ plus VC and VE treated groups, respectively. Consequently, the differences in the concentrations of $AFB_1$ related materials between vitamin treated and non-treated groups were significant. Immunohistochemistry revealed brownish infiltration of $AFB_1$ around central vein and sinusoid in $AFB_1-treated$ group. This manifestation was distinctly reduced in $AFB_1$ plus VC and VE treated groups.

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EXPRESSION OF OD314 DURING AMELOBLAST DIFFERENTIATION AND MATURATION (법랑모세포 분화와 성숙과정에서 OD314의 발현)

  • Park, Joo-Cheol;Ahn, Seong-Min;Kim, Heung-Joong;Jeong, Moon-Jin;Park, Min-Ju;Shin, In-Cheol;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.423-430
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    • 2005
  • Ameloblasts are responsible for the formation and maintenance of enamel which is an epithelially derived protective covering for teeth. Ameloblast differentiation is controlled by sequential epithelial-mesenchymal interactions. However, little is known about the differentiation and maturation mechanisms. OD314 was firstly identified from odontoblasts by subtraction between odontoblast/pulp cells and osteoblast/dental papilla cells, even though OD314 protein was also expressed in ameloblast during tooth formation. In this study, to better understand the biological function of OD314 during amelogenesis, we examined expression of the OD314 mRNA and protein in various stages of ameloblast differentiation using in-situ hybridization and immunohistochemistry. The results were as follows : 1. The ameloblast showed 4 main morphological and functional stages referred to as the presecretory, secretory, smooth-ended, and ruffle-ended. 2. OD314 mRNA was expressed in secretory ameloblast and increased according to the maturation of the cells. 3. OD314 protein was not expressed in presecretory ameloblast but expressed in secretory ameloblast and maturative ameloblast. OD314 protein was distributed in entire cytoplasm of secretory ameloblast. However, OD314 was localized at the proxiamal and distal portion of the cytoplasm of smooth-ended and ruffle-ended ameloblast. These results suggest that OD314 may play important roles in the ameloblast differentiation and maturation.

Induction of Apoptosis by Piceatannol in YD-15 Human Oral Cancer Cells (피세아타놀에 의한 YD-15 구강암세포의 세포자가사멸 유도 효과)

  • Lee, Hae-Nim;Jang, Hye-Yeon;Kim, Hyeong-Jin;Shin, Seong-Ah;Choo, Gang-Sik;Park, Byung-Kwon;Kim, Byeong-Soo;Jung, Ji-Youn
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.7
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    • pp.975-982
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    • 2015
  • Piceatannol (trans-3,4,3',5'-trihydroxystilbene), a natural stilbene, is an analogue of resveratrol. In the present study, possible mechanisms by which piceatannol exerts its pro-apoptotic action in cultured human oral cancer YD-15 cells were investigated. To investigate whether or not piceatannol has effects on cancer cell viability, human oral YD-15 cells were treated with piceatannol (0, 50, and $100{\mu}M$). Piceatannol treatment ($100{\mu}M$) showed the strongest inhibition of cell proliferation and reduced cell viability in a dose-dependent manner. Chromatin condensation detected by DAPI staining significantly increased in a concentration-dependent manner, indicating apoptosis. Piceatannol treatment activated initiator Bax (pro-apoptotic) and cPARP in a concentration-dependent manner. Further, piceatannol induced down-regulation of Bcl-2 (anti-apoptotic). We also evaluated the activity of piceatannol against oral cavity cancer tumors in mice. Piceatannol-treated nude mice bearing YD-15 xenograft tumors exhibited significantly reduced tumor volume and weight due to the potent effect of piceatannol on tumor cell apoptosis, as determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. Immunohistochemistry staining showed elevated expression of cleaved-caspase-3 as well as reduced expression of Ki-67 in the piceatannol-treated group. Therefore, piceatannol can be developed as a cancer preventive medicine due to its growth inhibitory effects and induction of apoptosis in human oral cancer cells.

Effect on bone healing by the application of low intensity pulsed ultrasound after injection of adipose tissue-derived stem cells at the implantation of titanium implant in the tibia of diabetes-induced rat (당뇨유도 백서 경골에 티타늄 임플란트 매식 시 지방조직 유래 줄기세포 주입 후 저출력 초음파 적용이 골치유에 미치는 영향)

  • Jung, Tae-Young;Park, Sang-Jun;Hwang, Dae-Suk;Kim, Yong-Deok;Lee, Soo-Woon;Kim, Uk-Kyu
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.4
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    • pp.301-311
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    • 2011
  • Introduction: This study examined the effect of the application of low intensity pulsed ultrasound on bone healing after an injection of adipose tissue-derived stem cells (ADSCs) during the implantation of a titanium implant in the tibia of diabetes-induced rats. Materials and Methods: Twelve Sprague-Dawely rats were used. After inducing diabetes, the ADSCs were injected into the hole for the implant. Customized screw type implants, 2.0 mm in diameter and 3.5 mm in length, were implanted in both the tibia of the diabetes-induced rats. After implantation, LIPUS was applied with parameters of 3 MHz, 40 mW/$cm^2$, and 10 minutes for 7 days to the left tibiae (experimental group) of the diabetesinduced rats. The right tibiae in each rat were used in the control group. At 1, 2 and 4 week rats were sacrificed, and the bone tissues of both tibia were harvested. The bone tissues of the three rats in each week were used for bone-to-implant contact (BIC) and bone area (BA) analyses and the bone tissues of one rat were used to make sagittal serial sections. Results: In histomorphometric analyses, the BIC in the experimental and control group were respectively, $39.00{\pm}18.17%$ and $42.87{\pm}9.27%$ at 1 week, $43.74{\pm}6.83%$ and $32.27{\pm}6.00%$ at 2 weeks, and $32.62{\pm}11.02%$ and $47.10{\pm}9.77%$ at 4 weeks. The BA in experimental and control group were respectively, $37.28{\pm}3.68%$ and $31.90{\pm}2.84%$ at 1 week, $20.62{\pm}2.47%$ and $15.64{\pm}2.69%$ at 2 weeks, and $11.37{\pm}4.54%$ and $17.69{\pm}8.77%$ at 4 weeks. In immunohistochemistry analyses, Osteoprotegerin expression was strong at 1 and 2 weeks in the experimental group than the control group. Receptor activator of nuclear factor kB ligand expression showed similar staining at each week in the experimental and control group. Conclusion: These results suggest that the application of low intensity pulsed ultrasound after an injection of adipose tissue-derived stem cells during the implantation of titanium implants in the tibia of diabetes-induced rats provided some positive effect on bone regeneration at the early stage after implantation. On the other hand, this method is unable to increase the level of osseointegration and bone regeneration of the implant in an uncontrolled diabetic patient.

Study on Effect of Saengbal-eum-II(Shēngfà-yĭn-II)on Hair Regrowth Promotion in C57BL/6 Mice (생발음(生髮飮)-II 피부도포가 C57BL/6 마우스의 육모촉진에 미치는 효과)

  • Han, Ae-Ri;Sohn, Nak-Won;Chung, Seok-Hee;Kim, Sung-Su;Song, Mi-Yeon
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.4
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    • pp.95-113
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    • 2009
  • Objectives : Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) is a hair care product which is composed of ten plant extracts used in oriental medicine. This study was carried out to investigate the effects of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) on hair regrowth and cytokine changes in a shaving model of C57BL/6 mice. Materials and Methods : Five-week-old mice were acclimated for 1 week at a temperature between $21-23^{\circ}C$, 40-60% relative humidity, and 12h of a light/dark cycle before beginning of the experiment. There were three experimental groups including 50% ethanol (EtOH, control), a positive control of 3% Minoxidil, and 30% Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) in 50% ethanol in 18 female mice. The test compounds were topically treated once a day over 12 days. The hair regrowth was photographically and histologically determined during the experimental period of 12 days. Revelation of EGF, $TGF-{\beta}1$ and IL-6 in hair follicle were also determined using immunohistochemistry. In addition to that, IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue were determined using enzyme-linked immunosorbent assay(ELISA). Results : Hair regrowth in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups was promoted earlier and faster than the control group. Concentrations of hairs and thick-hair ratio in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups were promoted than the control group. EGF was moderately positive in hair follicle of 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups, but negative in the control group. $TGF-{\beta}1$ was not significantly difference between the groups. IL-6 in hair follicle of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group was negative, but weakly positive in 3% Minoxidil and control group. IL-6 and $IL-1{\beta}$ in skin tissue were significantly decreased in Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group, but there was not significantly decreased in 3% Minoxidil and control group. $TNF-{\alpha}$ in skin tissue was significantly decreased in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups. Conclusions : These results suggest that Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) has hair growth promoting activity and it can be used for treatment of alopecia. And these effects relate to EGF revelation of hair follicle and a decrease IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue.

Gastrointestinal Stromal Tumor (GIST) of the Stomach: Clinicopathologic Analysis and Outcome (위에 발생한 위장관 간질성 종양의 임상병리학적 특성과 치료성적)

  • Ryu Je-Seock;Lee Sung-Ryul;Choi Sae-Byeol;Park Sung-Soo;Lee Ju-Han;Kim Seung-Joo;Kim Chong-Suk;Chae Yang-Seok;Mok Young-Jae
    • Journal of Gastric Cancer
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    • v.5 no.1
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    • pp.40-46
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    • 2005
  • Purpose: Gastrointestinal stromal tumors (GISTs) are mesenchymal neoplasms of the gastrointestinal tract. GISTs are positive for the expression of c-Kit protein at immunohistochemistry, and their clinical presentations vary. This retrospective study was performed to evaluate the clincopathologic characteristics of GISTs and to define the prognostic factors. Materials and Methods: 40 patients who underwent a complete resection of a GIST during the period $1996\~2003$ at the Department of Surgery, Korea University College of Medicine, were studied. We divided them into low- and high-risk. groups by using tumor size and mitotic count: 23 cases were low risk, and 17 were high risk. Clinicopathologic features, immunohistochemical findings, and prognoses were compared between the low- and the high-risk groups. Results: The mean age of the 40 patients was $61.3\pm11.1$years, and the male-to-female ratio was 1:1.1. There was no significant difference in age and sex between the groups. A comparative analysis revealed tumor size, mitotic count, clinical symptoms, preoperative pathologic diagnosis, ulceration, and necrosis to be variables that had statistically significant differences between the high- and the low-risk groups. In the univariate analysis, tumor size, mitotic count, ulceration, necrosis, and abnormal endoscopic ultrasound findings were associated with disease-free survival, but in the multivariate analysis, mitotic activity was the only independent factor associated with disease-free survival. 8 patients had recurrences during the follow-up period, and four of them were treated with STI-571 (imatinib mesylate, $Gleevec^{(R)}$). The treated patients have survived until now; however, two of non-treated patients died from disease progression. Conclusion: Based on this study, tumor size, ulceration, and necrosis are significant factors affecting survival, and mitotic activity may be a useful prognostic marker. STI-571 may be used in an adjuvant setting because the drug has shown anticancer activity in patients with recurrence or metastasis.

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The Immunohistochemical Analysis for the Expression of Survivin, an Inhibitor of Apoptosis Protein, in Non-small Cell Lung Cancer (비소세포폐암에서 아포프토시스 억제 단백질 Survivin 발현에 관한 면역조직학적 분석)

  • Ko, Mi-Hye;Myoung, Na-Hye;Lee, Jae-Whan;Cho, Eun-Mi;Park, Jae-Seuk;Kim, Keun-Youl;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.909-921
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    • 2000
  • Background : Defects in apoptotic signaling pathways play important role in tumor initiation, progression, metastasis and resistance to treatment. Several proteins which may promote tumorigenesis by inhibiting apoptosis were identified. The survivin protein is the member of inhibitor of apoptosis protein(IAPs) family which inhibits apoptosis. Unlike other IAPs, it is expressed in during the fetal period but not in adult differentiated tissues. Many reports have stated that survivin is selectively expressed in many cancer cell lines and cancer tissues. We performed immunohistochemical analysis for survivin expression in non-mall cell lung cancer to get evaluate its clinical implication. Methods : Twenty nine surgically resected lung cancers were examined. Immunohistochemical staining were performed by immuno-peroxidase technique using avidin-biotinylated horseradish pemxidase complex in the formalin-fixed, paraffin-embedded tissue $4{\mu}m$ section. Anti-survivin polyclonal antibody was used for primary antibody and anti-p53 monoclonal antibody was also used to analyze the correlation between survivin and p53 expression. The survivin expression scores were determined by as the sum of the stained area and intensity. Results : Immunohistochemical analysis showed cancer specific expression of survivin in 20 of 29 cases (69.0%). Western blot analysis also showed the selective survivin expression in tumor tissue. There was no correlation between survivin expression and clinicopathological parameters and prognosis. We analyzed the ∞π'elation between survivin expression and p53 expression, but found none. Conclusion: We confirmed the tumor specific expression of survival in non-small cell lung canær. But this expression was not correlated with clinical parameters as well as histology, tumor stage, recurrence, and survival rate. Also it was not statistically correlated with the expression of p53.

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Increased Expression of Phospholipase C-$\gamma1$ Activator Protein, AHNAK in Human Lung Cancer Tissues (인체 폐암조직에서 Phospholipase C-$\gamma1$의 활성화 단백, AHNAK의 발현양상)

  • Oh, Yoon-Jung;Park, Chun-Seong;Choi, So-Yeon;Cheong, Seong-Cheoll;Lee, Sun-Min;Hwang, Sung-Chul;Lee, Yi-Hyeong;Hahn, Myung-Ho;Lee, Kyi-Beom;Ryu, Han-Young;Ha, Mahn-Joon;Bae, Yoon-Su;Rhee, Seo-Goo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.3
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    • pp.347-355
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    • 1999
  • Background: Phospholipase C(PLC) plays a central role in cellular signal transduction and is important in cellular growth, differentiation and transformation. There are currently ten known mammalian isozymes of PLC reported to this date. Hydrolysis of phosphatidylinositol 4,5-bisphosphate($PIP_2$) by PLC produces two important second messengers, inositol 1,4,5-trisphosphate($IP_3$) and diacylglycerol. PLC-${\gamma}1$, previously, was known to be activated mainly through growth factor receptor tyrosine kinase. Other mechanisms of activating PLC-yl have been reported such as activation through tau protein in the presence of arachidonic acid in bovine brain and activation by $IP_3$, phosphatidic acid, etc. Very recently, another PLC-${\gamma}1$ activator protein such as tau has been found in bovine lung tissue, which now is considered to be AHNAK protein. But there has been no report concerning AHNAK and its associated disease to this date. In this study, we examined the expression of the PLC-${\gamma}1$ activator, AHNAK, in lung cancer specimens and their paired normal. Methods: From surgically resected human lung cancer tissues taken from twenty-eight patients and their paired normal counterparts, we evaluated expression level of AHNAK protein using immunoblot analysis of total tissue extract Immunohistochemical stain was performed with primary antibody against AHNAK protein. Results: Twenty-two among twenty-eight lung cancer tissues showed overexpression of AHNAK protein (eight of fourteen squamous cell lung cancers, all of fourteen adenocarcinomas). The resulting bands were multiple ranging from 70 to 200 kDa in molecular weight and each band was indistinct and formed a smear, reflecting mobility shift mainly due to proteolysis during extraction process. On immunohistochemistry, lung cancer tissues showed a very heavy, dense staining with anti-AHNAK protein antibody as compared to the surrounding normal lung tissue, coresponding well with the results of the western blot Conclusion: The overexpression of PLC-${\gamma}1$ activator protein, AHNAK in lung cancer may provide evidence that the AHNAK protein and PLC-${\gamma}1$ act in concerted manner in carcinogenesis.

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Immunomodulatory effects of β-1,3/1,6-glucan and lactic acid bacteria in LP-BM5 murine leukemia viruses-induced murine acquired immune deficiency syndrome (면역결핍 모델에서 β-1,3/1,6-glucan과 유산균을 이용한 in vivo 면역 활성 조절 효과)

  • Kim, Min-Soo;Kim, JoongSu;Ryu, Min Jung;Kim, Ki hong;Hwang, Kwontack
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1158-1167
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    • 2017
  • In this study, ${\beta}$-1,3/1,6-glucan, lactic acid bacteria, and ${\beta}$-1,3/1,6-glucan+lactic acid bacteria were tested for 10 weeks using an immunodeficient animal model infected with LP-BM5 murine AIDS virus On the immune activity. Cytokines production, plasma immunoglobulin concentration, T cell and B cell proliferation were measured. As a result, the T cell proliferative capacity which was weakened by immunization with LP-BM5 murine AIDS virus increased significantly T cell proliferative capacity compared with the red ginseng control group. B cell proliferative capacity was significantly higher than the infected control group. Increased B cell proliferation was reduced. In the cytokine production, IL-2, IL-12 and IL-15 in the Th1-type cytokine increased the secretion of IL-2, IL-12 and IL-15 compared to the infected control. The proliferative capacity of the treated group was higher than that of the mixed treatment group. TNF-${\alpha}$ was significantly decreased compared with the infected control group. The IL-4, IL-6 and IL-10 levels were significantly inhibited in the infected control group and the Th1/Th2 type cytokine expression was regulated by immunohistochemistry. IgE, IgA, and IgG levels were significantly lower in the immunoglobulin secretion assay than in the control. As a result, the immunomodulatory effect of ${\beta}$-1,3/1,6-glucan+lactic acid bacteria was confirmed by mixing with LP-BM5 murine AIDS virus-infected immunodeficient animal model.