• 대한본초학회지
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• 제20권2호
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• pp.35-42
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• 2005

Objective : This study was performed to investigate the effect of oral administration of Oldenlandiae Diffusae Herba (ODH) on eosinophil, immunoglobulin-E and interleukin-4 in the experimental asthma induced by ovalbumin. Methods : Asthma was induced to Balb/c mouse by i.p. injection and aerosol immunization with ovalbumin. It was observed the change of the eosinophil number in the BALF. And, it was observed the change of the concentrations of IL-4 in BALF and splenocyte, IgE in serum by ELISA. Results : 1. The number of eosinophil in BALF was significantly decreased in ODH group copared with control group. 2. Concentration of IL-4 in serum, BALF and splenocyte was significantly decreased in ODH group compared with control group, respectively. 3. Level of IgE in serum was significantly decreased in ODH group compared with control group. Conclusion : We found that the effect of ODH extract in asthma was implicated in reductions of IL-4 released from Th2 cell, and decreases of IgE from plasma cell. These findings suggest that ODH extract can produce anti-asthmatic effect, which may playa role in allergen-induced asthma therapy.

• 동의생리병리학회지
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• 제19권2호
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• pp.434-440
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• 2005

This study was conducted to find out the effect of Bojungikgitanggamibang(BIT) on growth, learning and memory of rats. The effects of BIT on learning and memory performance were examined in normal or memory impaired mice by using water maze task. Memory was impaired by 192 saporin. Body weight and growth of bone and tail of sample group were not significant compared with those of control groups. Acquisition test of water maze revealed that acqusitive ability of sample group significantly improved on 4,5th day compared with control group, while retentive ability of sample group was not significant. ChAT cell numbers of medial septum of sample group was significant compared with control group, and so was those of CA1, CA2 parts of hippocampus. On ChAT cell numbers of hippocampus, in CA2 part. These results suggest that BIT has an improving effect on the impaired learning through the effects on memory registration and retrieval.

• 대한약침학회지
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• 제5권1호
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• pp.61-79
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• 2002

Objective: The purpose of this study was to investigate acute and subacute toxicity and sarcoma-180 anti-cancer effects of Herbal acupuncture with Anneniacae amarum semen (Haeng-in) in mice and rats. Method: Balble mice were injected intraperitoneany with Haeng-In extract for $LD_{50}$ and acute toxicity test. Sprague-Dawley rats were injected intraperitoneally with Haeng-In extract for subacute toxicity test. TheAnneniacae amarum semen Herbal-Acupuncture was injected on Chung-wan (CV12) of mice with Sarcoma-180 cancer cell line. Results: 1. $LD_{50}$ was uncountable as none of the subjects expired from the treatment groups during the test. 2. The clinical signs and the body weight of mice treated with 0.1cc and 0.2cc Haeng-In extract were not affected during the acute toxicity test. 3. In acute toxicity test of serum biochemical values of mice, total protein and albumin were decreased in treatment group Ⅰ. Glucose was increased, and total cholesterol was decreased in treatment groups. GPT was increased in treatment group Ⅰ. 4. In subacute toxicity test, toxic symptoms were not detected in the treatment groups. 5. In subacute toxicity test, the body weight was increased in treatment groups on 14th and 21st day. 6. In subacute toxicity test. liver weight was increased in treatment group Ⅱ, and spleen weight was increased in treatment group Ⅱ. Lung weight was increased in an the treatment groups.(p<0.05) 7. In subacute toxicity test, severe tissue injury was found in lung and liver, especially treatment group Ⅰshowed more significant lung damage compared to treatment group l. 8. In subacute toxicity test, WBC. MCH and MCHC were increased in an the treatment groups, RBC, HGB and HCT were decreased in treatment group H(p<0.05). 9. In subacute toxicity test of serum biochemical values of rats, triglyceride was decreased in all the treatment groups. ALP was decreased in treatment group Ⅰ. and creatinine was decreased in treatment group Ⅱ. BUN/CR was increased in treatment group Ⅱ(p<0.05). 10. Median survival time of Sarcoma-180 cancer cell treated with Haeng-In was increased in all the treatment groups by twenty percent, compared to the control group(p<0.05). 11. Natural killer cell activity about the Sarcoma-180 cell was decreased at the ratio of 100:1 but was increased at the ratio of 10:1. In treatment group Ⅱ, increase was found at the ratio of 100:1 and 50:1 (p<0.05). 12. Interleukin-2 productivity of the Sarcoma-180 cell was decreased in treatment group I, but was increased in treatment group Ⅱ(p<0.05). Conclusion: According to the results, we can conclude Herbal-acupuncture with Anneniacae amarum semen caused toxicity, and had effects in Sarcoma-180 cancer cell.

• 한국동물학회지
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• 제32권4호
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• pp.337-350
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• 1989

본 연구에서는 SDS/polyacrylamide젤 전기영동과 2차원 전기영동을 사용하여 Gerris paludum(소금쟁이)의 난 형성 동안 난소 단백질과 혈림프 단백질을 비교하였다. 또한 난물질의 구성성분인 glycoprotein의 양상을 살펴 보았다. 결과는 다음과 같다. 난을 갖지 않는 5령의 난소 단백질의 양상을 성체의 산란 무 난소 단백질 퍼턴과 유사하였다. 난 발달동안의 난소의 단백질중 M.W 66,000-110,000 사이의 단백질이 난을 갖지 않은 난소 단백질에 비해 많은 양이 존재하였다. 난을 갖지 않은 난소에서는 glycoprotein은 나타나지 않았다. glycoprotein은 5령의 난 발달이 진행됩에 따라 점차적으로 증가하였다. 산란후 성체의 난소에서는 5령의 난을 갖지 않는 난소에서처럼 glycoprotein은 나타나지 않았다. 또한, M.W 66,000-110,000 사이의 혈림프 단백질은 5령의 난 형성 동안 양적으로 증가하였다. 이러한 결과는 난소 단백질 중 일부를 혈림프로부터 얻을 것이라는 사실을 제시하는 것이다. 발달하고 있는 난의 glycoprotein은 이 단계의 혈림프에서도 점차 증가하였다. 낮은 분자 영역에서 하나의 band (M.W. 9,500)가 난을 갖지 않는 5령과 성체의 혈림프에서 분리되었다. 이 단백질은 고분자량의 영역(M.W. 66,000-110,000)에서 나타나는 glycoprotein 전구체라고 생 각된다. 2차원 전기영동에서는 난을 갖지 않은 난소 단백질이 난을 가지고 있는 난소 단백질에 비해 적은 spots로 분리되었고 그 양도 적었다. M.W 45,000-110,000 사이의 band들은 2차원 전기영동에서 산성쪽으로 분리되어 나타났다. 특히, M.W. 102,000의 분자량을 가지는 band는 4개의 spots bl, b2, b3 및 b4로 분리되었다. 반면에 M.W 24,000이하의 단백질들은 산성쪽의 단백질보다 염기성쪽으로 분리되었다. Gepris paludum의 난황 형성과정에 있어서 세포내 기관(= 세포 소기관)의 기작이 난세포의 난황단백질 합성과 관계를 가지고 있었다.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.469-490
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• 1996

The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

• 대한한의학회지
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• 제23권4호
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• pp.113-124
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• 2002

Objects: This research was conducted to investigate the protective effect of Bupleuri Radix against ischemic damage using PC12 cells and global ischemia in gerbils, Methods: To observe the protective effect of Bupleuri Radixon ischemic damage, viability and changes in activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase and production of malondialdehyde (MDA) were observed after treating PC12 cells with Bupleuri Radix during ischemic damage. Gerbils were divided into three groups: a normal group, a 5-minute two-vessel occlusion (2VO) group and a Bupleun Radix administered group after 2VO. The CCAs were occluded by microclip for 5 minutes, Bupleuri Radix was administered orally for 7 days after 2VO. Histological analysis was performed on the 7th day. For histological analysis, the brain tissue was stained with 1 % of cresyl violet solution. Results: 1. Bupleuri Radix has a protective effect against ischemia in the CA1 area of the gerbil's hippocampus 7 days after 5-minute occlusion. 2. In the hypoxia/reperfusion model using PC12 cells, the Bupleuri Radix has a protective effect against ischemia in the dose of 0.2{\;}\mu\textrm{g}/ml,2{\;}\mu\textrm{g}/ml{\;}and{\;} 20{\;}\mu\textrm{g}/ml$. 3. Bupleuri Radix increased the activities of glutathione peroxidase and catalase. 4. The increased activity of superoxidedismutase (SOD) by ischemic damage might have been induced as an act of self-protection. This study suggests that Bupleuri Radix has some neuroprotective effect against neuronal damage following cerebral ischemia in vivo with a widely used experimental model of cerebral ischemia in Mongolian gerbils. Bupleuri Radix also has protective effect on a hypoxia/reperfusion cell culture model using PC12 cells. Conclusions: Bupleuri Radix has protective effect against ischemic brain damage during the early stages of ischemia.

• 한국수산과학회지
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• 제34권2호
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• pp.114-118
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• 2001

저염창란젓갈 제조 공정에 시 숙성공정에 교반과정을 도입하여 숙성기간의 단축과 제품의 균일화를 시도하였다. 먼저 시험구의 경우에는 원료창란을 연속교반 염장한 후 생성된 유출수를 제거하고 $0\pm2^{\circ}C$에서 4시간마다 10rpm으로 10분간 교반숙성하였으며, 대조구의 경우는 정치염장한 후 유출수를 포함하여 $0\pm2^{\circ}C$에서 정치숙성하였다. 60일간 숙성시키면서 이들의 품질변화를 측정한 결과 숙성일수가 경과할수록 pH는 낮아졌으며, 숙성 60일째에는 초기 pH 7.0에서 $6.2\~6.3$ 정도로 떨어졌으며 시험구가 대조구 보다 대체로 낮은 값을 나타내었다. 또한 시험구의 경우 숙성 30일째에Brix는 27.4, VBN은 $54.3 mg\%$ , 아미노태질소는 $87.9 mg\%$를 나타내었는데 비하여 대조구는 경우 숙성 50일째 Brix는 27.1, VBN은 $57.8 mg\%$, 아미노태질소는 $96.6mg\%$를 나타내었다. 숙성 중 미생물 변화는 대조구의 경우 50일째 $1.9\times10^6CFU/g$, 시험구의 경우 30일째 $2.6\times10^6CFU/g$으로 최대로 증가한 후 서서히 감소하는 경향을 보였다. 또한 관능검사 평가 결과도 대조구의 경우 50일째, 시험구의 경우 30일째에 최고값을 나타내어 창란젓갈의 최적 숙성기간은 대조구의 경우 50일, 시험구의 경우 30일로, 교반숙성 시키는 경우가 정치숙성에 비해 20일정도 공정일수를 단축할 수 있었다.

• 사상체질의학회지
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• 제27권2호
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• pp.270-287
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• 2015

Objectives To evaluate the neuroprotective effects of modified Yuldahanso-tang (MYH) in a Parkinson's disease mouse model. Methods 1) Four groups (each of 8 rats per group) were used in this study. 2) The neuroprotective effect of MYH was examined in a Parkinson's disease mouse model. C57BL/6 mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30 mg/kg/day), intraperitoneal (i.p.) for 5 days. 3) The brains of 2 mice per group were removed and frozen at $-20^{\circ}C$, and the striatum-substantia nigra part was seperated. The protein volume was measured by Bradford method following Bio-Rad protein analyzing kit. Using mouse/Rat Dopamine ELISA Assay Kit. 4) The brains of 2 mice per group were separated and removed. TH-immunohistochemical was examined in the MPTP-induced Parkinson's disease mice to evaluate the neuroprotective effects of MYH on ST and SNpc. 5) Two mice out of each group were anesthetized and skulls were opened from occipital to frontal direction to take out the brains. The brains added TTC solution for 20 minutes for staining. 6) The water tank used for morris water maze test was filled with $28^{\circ}C$ water, and a round platform of 10cm in diameter was installed for mice to step on. The study was carried out once a day within 30 seconds, keep exercising to step on the platform in the pool. 7) The brains of two mice out of each group were fixed in 10% formaldehyde solution and paraphillin substance was infiltrated. They were fragmented by microtome, and observed under an optical microscope after Hematoxylin & Eosin staining. 8) A round acrylic cylinder with its upper side open was filled with clean water and depressive mouse models were forced to swim for 15 minutes. After 24 hours the animals were put in the same equipment for 5 minutes and were forced to swim. 9) The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of Neurotransmitters in MPTP-MYH group. Results 1) MYH possess Dopamine cell protective effect on MPTP-induced injury in striatum and substantia nigra pars compacta. 2) MYH inhibits the loss of tyrosine hydroxylase-immunoreacitive (TH-IR) cells in the striatum and substantia nigra pars compacta on MPTP-induced injury in C57BL/6 mice. 3) MYH possesses improvement effect on MPTP-induced memory deterioration in C57BL/6 mice through the reduction of prolongated Sort of lost time by MPTP injection using the Morris water maze test. 4) MYH possesses hippocampal neuron protective effect on MPTP-induced injury in C57BL/6 mice. 5) MYH possesses improvement effect on MPTP-induced motor behaviour deficits and depression in C57BL/6 mice through the reduction of prolongated losing motion by MPTP injection using the Forced swimming test. 6) MYH increases serotonin product amount on MPTP-induced injury in C57BL/6 mice. Conclusions This experiment suggests that the neuroprotective effect of MYH is mediated by the increase in Dopamin, TH-ir cell, Hippocampus and Serotonin. Furthermore, MYH essential oil may serve as a potential preventive or therapeutic agent regarding Parkinson's disease.

• Archives of Plastic Surgery
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• 제38권4호
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• pp.438-444
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• 2011

Purpose: Adipose-derived stromal cells (ASCs) are readily harvested from lipoaspirated tissue or subcutaneous adipose tissue fragments. The stromal vascular fraction (SVF) is a heterogeneous set of cell populations that surround and support adipose tissue, which includes the stromal cells, ASCs, that have the ability to differentiate into cells of several lineages and contains cells from the microvasculature. The mechanisms that drive the ASCs into the osteoblast lineage are still not clear, but the process has been more extensively studied in bone marrow stromal cells. The purpose of this study was to investigate the osteogenic capacity of adipose derived SVF cells and evaluate bone formation following implantation of SVF cells into the bone defect of human phalanx. Methods: Case 1 a 43-year-old male was wounded while using a press machine. After first operation, segmental bone defects of the left 3rd and 4th middle phalanx occurred. At first we injected the SVF cells combined with demineralized bone matrix (DBM) to defected 4th middle phalangeal bone lesion. We used P (L/DL)LA [Poly (70L-lactide-co-30DL-lactide) Co Polymer P (L/DL)LA] as a scaffold. Next, we implanted the SVF cells combined with DBM to repair left 3rd middle phalangeal bone defect in sequence. Case 2 was a 25-year-old man with crushing hand injury. Three months after the previous surgery, we implanted the SVF cells combined with DBM to restore right 3rd middle phalangeal bone defect by syringe injection. Radiographic images were taken at follow-up hospital visits and evaluated radiographically by means of computerized analysis of digital images. Results: The phalangeal bone defect was treated with autologous SVF cells isolated and applied in a single operative procedure in combination with DBM. The SVF cells were supported in place with mechanical fixation with a resorbable macroporous sheets acting as a soft tissue barrier. The radiographic appearance of the defect revealed a restoration to average bone density and stable position of pharyngeal bone. Densitometric evaluations for digital X-ray revealed improved bone densities in two cases with pharyngeal bone defects, that is, 65.2% for 4th finger of the case 1, 60.5% for 3rd finger of the case 1 and 60.1% for the case 2. Conclusion: This study demonstrated that adipose derived stromal vascular fraction cells have osteogenic potential in two clinical case studies. Thus, these reports show that cells from the SVF cells have potential in many areas of clinical cell therapy and regenerative medicine, albeit a lot of work is yet to be done.

• IMMUNE NETWORK
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• 제5권2호
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• pp.78-88
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• 2005

Background: Collagen-induced arthritis (CIA) in mice is animal model of autoimmune disease known as rheumatic arthritis in human. We investigated CII-specific CD4+ T cell receptor usage in CIA mice. Methods: In CIA model, draining lymph node (dLN) CD4+ T cells and splenocytes at $3^{rd},\;5^{th},\;8^{th}$ week, we investigated CII-specific T cell proliferation, production of IL-17, IFN-${\gamma}$, TNF-${\alpha}$, IL-4 and IL-10. And we also performed anti-CII IgG Ab measurements in serum level, TCRV ${\beta}$ usage and T cell clonality with RT-PCR-SSCP analysis. Also, we performed proliferative response against CII when CII-specific T cell subset is deleted. Results: CIA mice showed more increase in the serum level of anti-CII IgG than normal mice after induction of arthritis. And the level of anti-CII IgG2a in CIA mice was increased after $3^{rd}$ week after primary immunization, while anti-CII IgG1 was decreased. Draining LN CD4+ T cells have proliferated against CII stimulation at $3^{rd}$ week after $1^{st}$immunization. CD4+T cells derived from dLN of CIA mice produced proinflammatory cytokine IFN-${\gamma}$, IL-17 etc. Draining LN CD4 T cells of CIA presented higher proportion of CD4+V ${\beta}3$+subset compared to those of normal mice at $3^{rd}$ week after $1^{st}$ immunization, and they were increased in proportion by CII stimulation. Draining LN CD4+ T cells without TCRV ${\beta}3+/V{\beta}8.1/8.2+/V{\beta}$10b+cells were not responsive against CII stimulation. But, CII-reactive response of TCRV ${\beta}3-/V{\beta}8.1/8.2-/V{\beta}$10b- T cells was recovered when $V{\beta}3+$ T cells were added in culture. Conclusion: Our results indicate that CD4+$V{\beta}3+$ T cells are selectively expanded in dLN of CIA mice, and their recovery upon CII re-stimulation in vitro, as well as the production Th1-type cytokines, may play pivotal role in CIA pathogenesis.