• Journal of Acupuncture Research
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• 제19권2호
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• pp.28-38
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• 2002

Objective : This study is aimed to investigate the effects of bee venom and purified bee venom on cell death in synovial cell line. Methods : It was evaluated by using MTT assay, morphological method, flow cytometry, immunocytochemistry analysis, RT-PCR. Results : The result obtained is as follows. 1. The MTT assay demonstrated that synovial cell viability was significantly inhibitted dose-dependently by treatment with BV and PBV in comparison with control. And the inhibitory effect of BV and PBV was almost same. 2. The morphologic study demonstrated that synovial cell showed apoptotic body resulted from apoptosis after treatment with BV and PBV for 6 hours using microscope. 3. The Flow cytometry demonstrated that apoptosis of synovial cell treated with BV and PBV was related with stop of cell cycle in stage of G0/G1. 4. Immunocytochemistry assay demonstrated that COX-II and iNOS were slightly expressed by treatment with BV and PBV in comparison with control group. 5. RT-PCR analysis demonstrated that COX-II were almost down-regulated by high dose treatment with BV and PBV in comparison with control group. iNOS were well down-regulated by treatment with $5{\mu}g/ml$ BV and PBV whereas it was well expressed in control group. Conclusion : These results suggest that bee venom and purified bee venom have significant effect on cell death in synovial cell line and further study is needed in vivo.

• Biomolecules & Therapeutics
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• 제25권6호
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• pp.618-624
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• 2017

Betulinic acid (BA), a natural pentacyclic triterpene found in many medicinal plants is known to have various biological activity including tumor suppression and anti-inflammatory effects. In this study, the cell-death induction effect of BA was investigated in BV-2 microglia cells. BA was cytotoxic to BV-2 cells with $IC_{50}$ of approximately $2.0{\mu}M$. Treatment of BA resulted in a dose-dependent chromosomal DNA degradation, suggesting that these cells underwent apoptosis. Flow cytometric analysis further confirmed that BA-treated BV-2 cells showed hypodiploid DNA content. BA treatment triggered apoptosis by decreasing Bcl-2 levels, activation of capase-3 protease and cleavage of PARP. In addition, BA treatment induced the accumulation of p62 and the increase in conversion of LC3-I to LC3-II, which are important autophagic flux monitoring markers. The increase in LC3-II indicates that BA treatment induced autophagosome formation, however, accumulation of p62 represents that the downstream autophagy pathway is blocked. It is demonstrated that BA induced cell death of BV-2 cells by inducing apoptosis and inhibiting autophagic flux. These data may provide important new information towards understanding the mechanisms by which BA induce cell death in microglia BV-2 cells.

• Journal of Acupuncture Research
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• 제23권2호
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• pp.139-157
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• 2006

In the present study, I have investigated the bee venom (BV) and melittin (a major component of BV) -mediated anti-proliferative effects, and defined its mechanisms of action in cultured rat aortic vascular smooth muscle cells (VSMCs). BV and melittin $(0.4{\sim}0.8\;{\mu}g/ml)$ effectively inhibited 50 ng/ml platelet derived growth factor BB (PDGF-BB)-induced VSMCs proliferations. The regulation of apoptosis has attracted much attention as a possible means of eliminating excessively proliferating VSMCs. In the present study, the treatment of BV and melittin strongly induced apoptosis of VSMCs. I examined the effects on $NF-{\kappa}B$ activation to investigate a possible mechanism for anti-proliferative effects of BV and melittin, the PDGF-BB-induced $I{\kappa}B{\alpha}$ phosphorylation and its degradation were potently inhibited by melittin, and DNA binding activity and nuclear translocation of $NF-{\kappa}B$ p50 subunit in response to the action of PDGF-BB were potently attenuated by melittin. In further investigations, melittin markedly inhibited the PDGF-BB-induced phosphorylation of Akt but not ERK1/2, upstream signals of $NF-{\kappa}B$. Treatment of melittin also potently induced pro-apoptotic protein p53, Bax, and caspase-3 expression, but decreased anti-apoptotic protein Bcl-2 expression. These results suggest that the anti-proliferative effects of BV and melittin in VSMCs through induction of apoptosis via suppressions of $NF-{\kappa}B$ and Akt activation, and enhancement of apoptotic signal pathway. Based on these results, BV acupuncture can be a candidate as a therapeutic method for restenosis and atherosclerosis.

• 대한약침학회지
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• 제3권1호
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• pp.65-87
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• 2000

This study was designed to evaluate the effect of the bee venom(BV) aqua-acupuncture on the neuronal activities of catecholaminergic(tyrosine hydroxylase : TH, dopamine ${\beta}$ hydroxylase : D${\beta}$H) system in the brainstem. After the BV aqua-acupuncture was applied on Chok-Samni(ST36) and the gluteal part(Blank locus) in rats. Also, the number of colocalization between catecholamine containing neurons and Fos immunoreactive neurons were analyzed by using the double immunohistochemical technique. The results of the experiments were summarized as follows : 1. In DR and LC, Chok-Samni group and the Blank locus group showed more significant increase in the number of colocalization between TH containing neurons and Fos immunoreactive neurons than the control group. Furthermore, Chok-Samni group showed more significant increase than the Blank locus group. Also, in Arc, Chok-Samni group showed more significant increase than the Blank locus group and the control group. 2. In LC, Chok-Samni group showed more significant increase in the number of colocalization between D${\beta}$H containing neurons and Fos immunoreactive neurons than the Blank locus group and the control group. Also, in A5, Chok-Samni group and the Blank locus group showed more significant increase than the control group. Chok-Samni group showed more significant increase than the Blank locus group. However, there was no significant change in A7. Consequently, the BV aqua-acupuncture increased more potent the number of Fos immunoreactive neurons and the activity of catecholaminergic neurons. Furthermore, the BV aqua-acupuncture was more effective on Chok-Samni than Blank locus group. These results indicate that the BV aqua-acupuncture is very effective therapy to control pain. The therapeutic effect of BV aqua-acupunture may associated with the endogenous modulatory system such as catecholamine. Those data from the study can be applied to establish the effective treatment of the BV for pain control in the clinical field.

• 대한약침학회지
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• 제7권1호
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• pp.63-69
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• 2004

Objective : This study was designed to find out whether NS(normal saline) is able to be constituted as an appropriate control group for CC(Cervi Cornu Parvum herbal acupuncture), CF(Carthami-Flos herbal acupuncture) and BV(bee venom herbal acupuncture). Method : NS and three herbal acupuncture were inserted into Quchi(LI 11) of the subjects. After 5 minutes the subjects completed a questionnaire rating the intensity of 21 kinds of acupuncture sensation; hurting, penetrating, sharp, aching, intense, spreading, radiating, tingling, pricking, stinging, pulling, heavy, dull, numb, electric, shocking, hot, burning, cool, pulsing, and throbbing. We compared subjective evaluations of acupuncture sensation between or among the groups. Results : As for CC half items of the acupuncture sensation were significantly different from NS. As for CF all items were not significantly different from NS. As for CC all items were significantly different from NS except one item. In general the score of CF acupuncture sensation was lower than the others and the score of BV acupuncture sensation was higher than the others from comparison of sensation among herbal acupunctures(CC, CF, BV). Conclusion : We found that NS is able to be an appropriate placebo herbal acupuncture for CF. Further study is needed for new placebo herbal acupuncture for CC and BV.

• 대한임상검사과학회지
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• 제52권4호
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• pp.389-397
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• 2020

Microglial cells function as major immune cells in the brain, playing an important role in the protection and damage of neurons. BV2 microglia, activated by drug stimulation, secrete inflammatory cytokines by activating the nuclear factor kappa-light-chain-enhancer of the activated B cells pathway and are involved in neuroinflammatory and immune responses. The overactivation of microglia by stimuli can cause neuronal damage, leading to brain disease. Noni, a natural product, reduces the activity of microglia to prevent neuronal damage and is a potential natural medicine because it exerts excellent regeneration and anti-inflammatory effects on damaged cells. In this study, when noni was used to treat BV2 cells stimulated by the anti-cancer drug doxorubicin, it reduced the release of pro-inflammatory cytokines from BV2. On the other hand, neuronal damage is a side effect of doxorubicin. Therefore, the cytokines released from doxorubicin-stimulated BV2 cells treated with noni had a positive effect on the neuronal viability compared to those released from doxorubicin-stimulated BV2 cells not treated with Noni. Thus, Noni increases neuronal viability. These results suggest that noni inhibits the release of cytokines by regulating the nuclear factor kappa-light-chain-enhancer of the activated B cells pathway of BV2, thereby inhibiting neuronal damage.

• Imaging Science in Dentistry
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• 제52권2호
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• pp.171-179
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• 2022

Purpose: The aim of this study was to assess the accuracy of cone-beam computed tomographic (CBCT) images obtained using different voxel sizes in measuring trabecular bone microstructure in comparison to micro-CT. Materials and Methods: Twelve human skull bones containing posterior-mandibular alveolar bone regions were analyzed. CBCT images were obtained at voxel sizes of 0.075mm(high: HI) and 0.2mm(standard: Std), while microCT imaging used voxel sizes of 0.06 mm (HI) and 0.12 mm (Std). Analyses were performed using CTAn software with the standardized automatic global threshold method. Intraclass correlation coefficients were used to evaluate the consistency and agreement of paired measurements for bone volume (BV), percent bone volume (BV/TV), bone surface (BS), trabecular thickness (TbTh), trabecular separation (TbSp), trabecular number (TbN), trabecular pattern factor(TbPf), and structure model index (SMI). Results: When compared to micro-CT, CBCT images had higher BV, BV/TV, and TbTh values, while micro-CT images had lower BS, TbSp, TbN, TbPf, and SMI values (P<0.05). The BV, BV/BT, TbTh, and TbSp variables were higher with Std voxels, whereas the BS, TbPf, and SMI variables were higher with HI voxels for both imaging methods. For each imaging modality and voxel size evaluated, BV, BS, and TbTh were significantly different(P<0.05). TbN, TbPf, and SMI showed statistically significant differences between imaging methods(P<0.05). The consistency and absolute agreement between micro-CT and CBCT were excellent for all variables. Conclusion: This study demonstrated the potential of high-resolution CBCT imaging for quantitative bone morphometry assessment.

• Journal of Acupuncture Research
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• 제25권2호
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• pp.151-164
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• 2008

Objectives : This study was designed to compare the effects on Korean Western Ontario and McMaster Universities Osteoarthritis Index(KWOMAC), 36-ltems Short-Form Health Survey(SF-36), Visual Analog Scale(VAS) and Nine Point Scale between 'Intramuscular Bee Venom Herbal Acupuncture' and 'Intracutaneous Bee Venom Herbal Acupuncture' in knee osteoarthritis patients for 4weeks. Methods : All the voluntary subject were diagnosed as knee osteoarthritis. Patients were randomly assigned either to two groups : Intramuscular bee venom herbal acupuncture treatment group(IM-BV group) or Intracutaneous bee venom herbal acupuncture treatment group(IC-BV group). The IM-BV group received bee venom herbal acupuncture intramuscularly on muscles closest to the pathologic site on ultrasonography. The IC-BV group received bee venom herbal acupuncture intracutaneously on $LE_{110}$, $LE_{201}$, $ST_{35}$, $ST_{36}$ and $LR_8$. Bee venom herbal acupuncture was applied twice a week for 4 weeks by an oriental medical doctor at Kyung-hee Oriental Medical Hospital. Both groups were checked of pathologic site by ultrasonography before treatment and follow up after four-week treatment. The patients were assessed by KWOMAC and SF-36 before and after four-week treatment. The changes in patients' pain was measured by VAS before treatment and after one-, two-, three- and four-week treatment. The patients' overall treatment outcome assesment was measured by Nine Point Scale after four-week treatment. Results : The results were as follows ; 1. The IM-BV group showed significant improvement on total, pain, stiffness and physical function in KWOMAC, and the IC-BV group showed significant improvement on total and physical function in KWOMAC after 4weeks compared to the pre-treatment. In KWOMAC, the IM-BV group showed more improvement on average compared to the IC-BV group, but there were no significant difference between two groups. 2. The IM-BV group showed significant improvement on VAS after one-week treatment. The IC-BV group showed significant improvement on VAS after two-week treatment. But there were no significant difference between two groups. 3. The IM-BV group showed significant improvement on physical functioning(PF) and bodily pain(BP) in SF-36, and the IC-BV group showed significant improvement on vitality(VT) and bodily pain(BP) in SF-36 after 4weeks compared to the pre-treatment. But there were no significant difference between two groups. 4. 88.2% of IM-BV group and 93.3% of IC-BV group graded their improvement as 'Fair' or better on Nine Point Scale. But there were no significant difference between two groups. Conclusions : This study suggests that in the treatment of knee osteoarthtitis patients, effects of 'Intramuscular Bee Venom Herbal Acupuncture' were not different from 'Intracutaneous Bee Venom Herbal Acupuncture' statistically. But both treatment showed effects on pain and physical function in knee osteoarthtitis patients.

• 대한한방내과학회지
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• 제30권4호
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• pp.845-857
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• 2009

Background and Objectives : Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) and bee venom (BV) have been reported to induce apoptosis in various cancer cell lines in vitro and to show antitumor activity against a variety of tumors in animal models. However, the comparative effect of VCA and BV on the anti-cancer effect and mechanisms of action has not been determined. In this study, the effect in a human hepatocellular carcinoma cell line, Hep G2 cells, was examined. Methods : Cytotoxic effects of VCA and BV on Hep G2 cells were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay in litro. The apoptotic cell death was then confirmed by propidium iodide staining and DNA fragmentation analysis. The mechanisms of action were examined by the expression of anti-apoptotic proteins and activation of mitogen-activated protein kinases. The involvement of kinase was examined in VCA or BV-induced apoptosis by using kinase inhibitors. Results : VCA and BV killed Hep G2 cells in a time and dose-dependent manner. Treatment of Hep G2 cells with VCA activated poly (ADP-ribose) polymerase-1 (PARP-1) known as a marker of apoptosis, and mitogen-activated protein kinases signaling pathways including MAPK/ERK, p38 MAPK and JNK. BV also activated PARP-1, MAPK/ERK. and p38 MAPK but not JNK. The expression level of anti-apoptotic molecule, Bcl-X, was decreased by VCA treatment but not by BV. Finally, the phosphorylation level of ERM proteins involved in the cytoskeleton homeostasis was decreased by both stimuli. VCA-induced apoptosis was partially inhibited by in the presence of JNK and p38 inhibitor, but BV only by p38 inhibitor. Conclusions : VCA-induced apoptosis is dependent on the activation of p38 and JNK. while BV-induced apoptosis is mediated by p38 activation in Hep G2 cells.

• 농약과학회지
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• 제12권3호
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• pp.283-290
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• 2008

베큘로바이러스는 일부 나비목 해충을 대상으로 방제하는 데 사용되고 있다. 그러나 화학농약에 비해 느린 살충효과 및 좁은 적용 해충으로 응용 범위에 한계를 갖고 있다. 본 연구는 이러한 한계를 극복하고자 곤충의 면역억제을 통해 바이러스 병원력을 제고시킬 수 있는 기술을 소개한다. 폴리드나바이러스는 일부 맵시벌 및 고치벌에 공생하는 곤충 DNA 바이러스 분류군이다. 프루텔고치벌(Cotesia plutellae) 유래 CpBV(Cotesia plutellae bracovirus)는 브라코바이러스에 속한 폴리드나바이러스로서 면역어제를 발휘하는 여러 유전자를 함유하고 있다. 이 가운데 7개의 CpBV유전자를 선발하고 이를 야생형Autographa California multiple nucleopolyhedrovirus(AcNPV)에 재조합하였다. 이들 재조합 베큘로바이러스를 이용하여 파밤나방(Spodoptera exigua)과 배추좀나방(Plutella xylostella)을 대상으로 생물 검정한 결과, 이들 대부분은 야생형의 바이러스와 유사하거나 우수한 살충력을 나타냈다. 특히 CpBV-ELP를 포함한 재조합 베큘로바이러스가 대조바이러스에 비해 살충시간을 약 2 일 이상단축시킴으로 가장 우수하였다. 이 재조합 베큘로바이러스는 농도에 따른 살충력증가와 배추를 가해하는 파밤나방을 대상으로 한 바이러스 살포 처리가 뚜렷한 방제효과를 나타내어 현장 적용 가능성을 제시하였다. 또한 본 연구는 이 재조합 바이러스의 살충력 제고 현상을 CpBV-ELP의 항바이러스 기작 억제라는 측면에서 고찰했다.