• Title/Summary/Keyword: Tissue growth

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Ginsenoside R $b_2$ and Rc Formation and Inorganic Elements Uptake in Ginseng Hairy Roots Cultures (인삼모상근 배양에서 Ginsenoside R $b_2$ 및 Rc 생성과 무기이온 흡수)

  • 양덕조;윤길영;최규명;유승희
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.461-468
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    • 2000
  • For the enhancement of ginsenoside production in hairy roots cultures of Panax ginseng, the uptake rate of inorganic elements and ginsenoside contents were investigated by different concentrations of about phosphorus (P $O_{4}$$^{[-10]}$ ) and nitrogen (N $H_{4}$$^{+}$, N $O_{3}$$^{[-10]}$ ) sources. According to increased phosphorus and nitrogen sources, the uptake rate of $Mg^{2+}$ and F $e^{2+}$ in ginseng hairy roots were significantly increased. The uptake rate of F $e^{2+}$ in 5.15 mM N $H_{4}$$^{+}$ was higher at 47.5% than that in 20.6 mM, whereas that of C $u^{2+}$ in 10.3 mM were higher at 123.1% than that in 41.2 mM. These results indicated that phosphorus and nitrogen sources act not only elevated growth of hairy roots but also the uptake-enhancement of the irons and other ions. The optimum concentration of phosphorus and nitrogen sources for the contents of free sugars were different to kinds of free sugars. The optimum concentration of phosphorus and nitrogen sources for the ginsenoside formation in ginseng hairy roots cultures were highest at the most low concentration of all. The contents of ginsenoside-R $b_2$and -Rc in 0.31 mM P $O_{4}$$^{[-10]}$ were increased to 44.7% and 29.9% than that in 0.62 mM P $O_{4}$$^{[-10]}$ , respectively. The contents of ginsenoside-R $b_2$ and -Rc in 5.15 mM N $H_{4}$$^{+}$ were increased to 21.7% and 31.9% than that in 10.30 mM N $H_{4}$$^{+}$, respectively. The contents of ginsenoside-R $b_2$and -Rc in 4.7 mn N $O_{3}$$^{[-10]}$ were also increased to 17.6% and 25.5% than that in 9.4 mM N $O_{3}$$^{[-10]}$ , respectively. These results indicate that enhancement of the ginsenoside formation in ginseng hairy roots was feasible by new medium modulation of concentration of phosphorus and nitrogen sources.rogen sources.

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Plant regeneration and transformation of grape (Vitis labrusca L.) via direct regeneration method (포도 (Vitis labrusca L.)의 직접 재분화 방법을 이용한 식물체 재분화와 형질전환)

  • Kim, Se Hee;Shin, Il Sheob;Cho, Kang Hee;Kim, Dae Hyun;Kim, Hyun Ran;Kim, Jeong Hee;Lim, Sun-Hyung;Kim, Ki Ok;Lee, Hyang Bun;Do, Kyung Ran;Hwang, Hae Seong
    • Journal of Plant Biotechnology
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    • v.40 no.4
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    • pp.210-216
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    • 2013
  • Efficient regeneration methods and transformation system are a priority for successful application of genetic engineering to vegetative propagated plants such as grape (Vitis labrusca L.). This research is to establish shoot regeneration system from plant explants for 'Campbell Early', 'Tamnara', 'Heukgoosul', 'Heukbosek' using two types of plant growth regulators supplemented to medium. The highest adventitious shoot regeneration rate of 5% was achieved on a medium containing of Murashige and Skoog (MS) inorganic salts and Linsmaier and Skoog (LS) vitamins, 2 mg/L of TDZ and 0.1 mg/L of IBA. Leaf tissue of 'Campbell Early', was co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, gus gene as reporter gene and resistance to kanamycin as selective agent, the other Agrobacterium strains, GV3101 containing the vector pB7 WG2D carrying with mPAP1-D gene. mPAP1-D is a regulatory genes of the anthocyanin biosynthetic pathway. 'Campbell Early' harboring mPAP1-D gene was readily able to be selected by red color due to anthocyanin accumulation in the transformed shoot. These results might be helpful for further studies to enhance the transformation efficiency in grape.

STUDIES ON THE CYSTINE COMPONENT IN THE SERICULTURAL PROTEINS OF BOMBYX MORI L. (가잠사단백질의 각과정에서의 Cystine 성분에 대한 연구)

  • Choe, Byong-Hee
    • Journal of Sericultural and Entomological Science
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    • v.2
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    • pp.1-31
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    • 1962
  • The purpose of this treatise is to prove the presence of cystine in silk fiber through wide sampling throughout all the sericultural processes of Bombyx mori.; also to show that disulfide cross linkages exist in the silk fiber. The conclusions reached were as follows: 1. Crystalline cystine was obtained from silk fibroin using Folin's Method. 2. Analytical data showing the cystine content of silk fiber and its related materials were obtained using Sullvan's Method as follows: Material Percent Cystine A. Mulberry leaf protein 0.175 B. Silkworm egg 0.33 C. Silkworm Body, matured, fat extracted, without silk gland 0.41 D. Silk gland, matured 1.23 E. Silkworm feces none F. Silkworm pupa, fat extracted 0.30 G. Silkworm moth, fat extracted 0.60 H. Raw Silk 0.22 I. Fibroin 0.175 J. Sericin 0.30 3. The presence of cystine in the silkworm was substantiated the existence of 0.175 % methionine in mulberry leaves and 0.12% methionine in the silk gland. 4. Part of the sulfhydryl compounds in the silk gland is believed to transfer to serine and methionine, with the former being secreted into the liquid silk finally as silk fiber and the latter used for nutritive purposes in the growing of silk gland tissue. 5. The cystine content is variable by mulberry species, silkworm species, sex, breeding process, and other culturing environments. 6. Hybrid silkworms require more nutritive amino acids for effective growth than the original parents, and secrete less of them as silk fiber. 7. From such an observation, the amino acid composition of silk fiber is believed to be fairly flexible. Cystine if included in the amorphous part of the fiber, especially in sericin. 8. The result from enriching the silkworm diet with pure cystine or wool cystine did not result in any advantage, therefore it is believed that the natural cystine and methionine contents in the mulberry leafaregoodenoughforsilkwormnutrition. 9. The disulfide cross linkage in silk fiber was verified by using the Harris Method. Contraction took place following the treatment of the fiber with various salts and acids. Comparisons were made with wool fiber. 10. During these experiments, the fibrious structure of silk fiber and the net-globular liquid form were photographed microscopically. It is believed that the globules of liquid silk are net-formed by the inter attraction of the OH ion of the globular peptide and the H ion of water as shown by the hair cracking behavior of the film. The net-globular protein precipitation from the mulberry protein solution showed that mulberry is a proper diet for the formation of fibrous protein in the silk fiber. 11. The significance of the presence of cystine in silk fiber as emphasized in this paper should result in modification of the general conception that cystine is absent from this fiber. NOTICE: A part of this treatise was presented at the annual Korea Sericultural Society meeting held in 1961.

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The Correlation between Bronchoscopic Morphology and Pathologic Type in Bronchogenic Carcinoma (폐암의 기관지 내시경 소견과 세포형의 연관성에 관한 연구)

  • Lee, Hyuk-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.4
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    • pp.389-396
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    • 1994
  • Background : The two most important purposes of fiberoptic bronchoscopy in lung cancer patients are obtaining tissue diagnosis and staging. The direct sign of lung cancer on FOB includes visible tumor, with smooth or nodular surface, with or without necrosis and infiltration. Variant cell types of lung cancer have their characteristic biological behaviors respectively. For example, squamous cell carcinoma grows slowly, invades locally and has easy necrosis resulting in cavitation, whereas adenocarcinoma shows early metastasis, small cell carcinoma shows rapid growth and higher early metastasis rate. Based on this, it could be hypothesized that each cell type may have characteristic bronchoscopic finding. Method : To answer this question, we reviewed 106 cases which were diagnosed as primary lung cancer and had bronchoscopically visible specific cancerous lesions. Results : The results were as follows. 1) Squamous cell carcinoma accounted for 66 cases(62.2%), adenocarcinoma 15 cases(14.2%), large cell carcinoma 3 cases(2.8%). 2) The endobronchial tumor lesion was arbitrarily classified into 5 types according to gross characteristics. Type A, multilobulating mass with necrosis, accounted for 24.5%, type B, multilobulating mass without necrosis, 25.5%, type C, round beefy mass, 9.4%, type D, infiltration with mucosal irregularity, 6.6%, and type E, infiltration without mucosal irregularity, 34%. 3) The analysis of correlation between endobronchial tumor pattern and specific cell type revealed that squamous cell carcinoma had relation with the morphologic type B and small cell carcinoma had relation with the morphologic type E, but adenocarcinoma had no preponderance in morphologic type. The gross appearance had influence on the diagnostic yields of biopsies and the diagnostic yields of lobulating mass types(type A, B) were higher than those of other types. Conclusion : From the above observations, it could be concluded that squamous cell carcinoma and small cell carcinoma have relations with specific types of bronchoscopic morphology, but not the case in adenocarcinoma.

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Expression of COX-2 and IDO by Uteroglobin Transduction in NSCLC Cell Lines (비소세포폐암 세포주에서 Uteroglobin Transduction이 COX-2 및 IDO의 발현에 미치는 영향)

  • Park, Gun Min;Lee, Sang-Min;Yim, Jae-Joon;Yang, Seok-Chul;Yoo, Chul Gyu;Lee, Choon-Taek;Han, Sung Koo;Sim, Young-Soo;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.66 no.4
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    • pp.274-279
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    • 2009
  • Background: Uteroglobin (UG) is a secretary protein that has strong immunomodulatory properties, and which is synthesized in most epithelia including lung tissue. Overexpression of UG is associated with decreased expression of cyclooxygenase (COX)-2 and suppression of cancer cell growth. Indoleamine 2,3-dioxygenase (IDO) catalyzes tryptophan along the kynurenine pathway, and both the reduction in local tryptophan and the production of tryptophan metabolites contribute to the immunosuppressive effects of IDO. Methods: In this study, we investigated the pattern of expression of COX-2 and IDO, and the effect of UG transduction in the expression of COX-2 and IDO in several non-small cell lung cancer cell lines, especially A549. Results: Both COX-2 and IDO were constitutionally expressed in A549 and H460 cells, and was reduced by UG transduction. In A549 cells, the slightly increased expression of COX-2 and IDO with the instillation of interferon-gamma (IFN-$\gamma$) was reduced by UG transduction. However, the reduced expression of COX-2 and IDO by UG transduction was not increased with IFN-$\gamma$ instillation in A549 cells. In both the A549 COX-2 sense and the A549 COX-2 anti-sense small interfering RNA (siRNA)-transfected cells, IDO was expressed; expression was reduced by UG transduction, irrespective of the expression of COX-2. Conclusion: The results suggest that the anti-proliferative function of UG may be associated with the immune tolerance pathway of IDO, which is independent of the COX-2 pathway.

Radiation Effect of X-Ray and Thermal Neutron on Robinia pseudoacacia L. and Some Other Species (아까시나무외 몇 수종(樹種)에 대(對)한 X-Ray와 Thermal Neutron의 조사효과(照射効果))

  • Kim, Chung Suk;Lee, Suk Koo;Hyun, Sin Kyu
    • Journal of Korean Society of Forest Science
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    • v.17 no.1
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    • pp.1-15
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    • 1973
  • In an effort to improve the major tree species in Korea, the seed of Robinia pseudoacacia, Pinus rigida, Pinus densiflora, Pinus thunbergii and Larix leptolepis were treated with X-ray and thermal neutron at the Brookhaven National Laboratory, and germination rate of the seed and some characteristics of the seedlings from irradiated seed were investigated and the results were summarized as follows. 1. The germination rate of the irradiated seed of Robinia pseudoacacia, Pinus densiflora, Pinus thunbergii and Pinus rigida was decreased, when the irradiation time of thermal neutron increased from 3 hours to 9 hours. The seed of Larix leptolepis was completely died out in all range of irradiation time. 2. The seed of Pinus densiflora, Robinia pseudoacacia and Pinus rigida showed low germination rate, when the dosage of radiation increased in the range of 10,000r-30,000r X-ray. This dosage of radiation was almost lethal to the seed of Pinus thunbergii and Larix leptolepis. 3. The growth rate of radiated Robinia pseudoacacia has been decreased when the dosage of X-ray and thermal neutron increased. However, the trees treated with thermal neutron for 3 hours showed 14.9 percent-increase in seedling height and some thornless individuals appeared in this treatment. 4. Individuals with variegated leaf, rugose leaf and albino were appeared in X-ray and thermal neutron treatment. 5. Abnormal mitosis of somatic cell, cell with two nucleoli, cell with two nuclei and chromosome clump in mitosis of somatic cell were observed in Robinia pseudoacacia irradiated with thermal neutron. 6. Resistanty against pawdery mildew was decreased in Robinia pseudoacacia radiated with X-ray and thermal neutron. 7. Length of stomata did not show any difference however number of stomata per unit area decreased in Robinia pseudoacacia radiated with thermal neutron. The leaves of Robinia pseudoacacia radiated with thermal neutron were thicker than those of non-treated one, but width of palisade tissue was decreased. The most sensitive one among those species to the thermal neutron treatment was Larix leptolepis, followed by Pinus densiflora, Robinia pseudoacacia, Pinus thunbergii and Pinus rigida in the order. In X-ray treatment, the most sensitive one was Larix leptolepis, followed by Pinus densiflora, Pinus thunbergii, Pinus rigida and Robinia pseudoacacia in the order. Morphological, cytological variation of the radiated Robinia pseudoacacia seemed to indicate some possibility to be used for tree improvement.

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3-Dimensional Micro-Computed Tomography Study on Bone Regeneration with Silk Fibroin, rh-Bone Morphogenetic Protein Loaded-Silk Fibroin and Tricalcium Phosphate Coated-Silk Fibroin in Rat Calvaria Defect

  • Pang, Eun-O;Park, Young-Ju;Park, Su-Hyun;Kang, Eung-Sun;Kweon, Hae-Yong;Kim, Soeng-Gon;Ko, Chang-Yong;Kim, Han-Sung;Nam, Jeong-Hun;Ahn, Jang-Hun;Chun, Ji-Hyun;Lee, Byeong-Min
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.34 no.1
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    • pp.1-11
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    • 2012
  • Purpose: The purpose of this study was to evaluate the bone regeneration capacity of silk fibroin (SF) when combined with beta tricalcium phosphate (${\beta}$-tricalcium phosphate [TCP]) and rh-bone morphogenetic protein (BMP) in vivo by micro-computed tomography (CT), soft x-ray, and histological analysis. Methods: A total of 56 critical size defects formed by a trephine bur made on 28 adult female Spague-Dawley rats were used for this study and the defect size was 5.0 mm in diameter. The defects were transplanted with (1) no graft material (raw defect), (2) autogenous bone, (3) SF ($10{\mu}g$), (4) SF-BMP ($10{\mu}g$, $0.8{\mu}g$ each), and (5) SF+${\beta}$-TCP ($10{\mu}g$). At 4 and 8 weeks after operation, the experimental animals were sacrificed. Samples were evaluated with soft x-ray, histological examinations and 3-dimensional micro-CT analysis. Results: In the 3-dimensional micro-CT evaluation, bone volume and bone surface data were higher in the SF-BMP ($12.8{\pm}1.5$, $138.6{\pm}45.0$ each) (P<0.05) and SF-TCP ($12.3{\pm}1.5$, $144.9{\pm}30.9$ each) group than in the SF group ($6.1{\pm}3.3$, $77.2{\pm}37.3$ each) (P<0.05), except for the autogenous group ($15.0{\pm}3.0$, $190.7{\pm}41.4$ each) at 4 weeks. At 8 weeks, SF-BMP ($16.8{\pm}3.5$, $173.9{\pm}34.2$ each) still revealed higher (P<0.05) bone volum and surface, but SF-TCP ($11.3{\pm}1.5$, $1132.9{\pm}52.1$ each) (P=0.5, P=0.2) revealed the same or lower amount compared with the SF group ($13.8{\pm}2.7$, $127.5{\pm}44.8$ each). The % of bone area determined by radiodensity was higher in the SF-TCP ($31.4{\pm}9.1%$) and SF-BMP ($36.2{\pm}16.2%$) groups than in the SF ($19.0{\pm}10.4$) group at the period of 4 weeks. Also, in the histological evaluation, the SF-BMP group revealed lower inflammation reaction, lower foreign body reaction and higher bone healing than the SF group at postoperative 4 weeks and 8 weeks. The SF-TCP group revealed lower inflammation at 4 weeks, but accordingly, as the TCP membrane was absorbed, inflammatory and foreign body reaction are increased at 8 weeks. Conclusion: The current study provides evidence that the silk fibrin can be used as an effective grafted material for tissue engineering bone generation through a combination of growth factor or surface treatment.

CORRELATION BETWEEN VASCULAR ENDOTHELIAL GRWOTH FACTOR SIGNALING AND MINERALIZATION DURING OSTEOBLASTIC DIFFERENTIATION OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골세포 분화과정에서 골기질 형성정도와 혈관내피세포성장인자 신호와의 상관관계)

  • Park, Bong-Wook;Byun, June-Ho;Ryu, Young-Mo;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.3
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    • pp.197-205
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    • 2007
  • Angiogenesis is a essential part for bone formation and bone fracture healing. Vascular endothelial growth factor (VEGF), one of the most important molecules among many angiogenic factors, is a specific mitogen for vascular endothelial cells. VEGF-mediated angiogenesis is required for bone formation and repair. However, the effect of VEGF on osteoblastic cells during osteogenesis is still controversial. In recent days, substantial progress have been made toward developing tissue-engineered alternatives to autologous bone grafting for maxillofacial bony defects. Periosteum has received considerable interest as a better source of adult stem cells. Periosteum has the advantage of easy harvest and contains various cell types and progenitor cells that are able to differentiate into a several mesenchymal lineages, including bone. Several studies have reported the bone formation potential of periosteal cells, however, the correlation between VEGF signaling and cultured human periosteal cell-derived osteogenesis has not been fully investigated yet. The purpose of this study was to examine the correlation between VEGF signaling and cultured human periosteal-derived cells osteogenesis. Periosteal tissues of $5\;{\times}\;20\;mm$ were obtained from mandible during surgical extraction of lower impacted third molar from 3 patients. Periosteal-derived cells were introduced into the cell culture and were subcultured once they reached confluence. After passage 3, the periosteal-derived cells were further cultured for 42 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and ${\beta}-glycerophosphate$. We evaluated the alkaline phosphatase (ALP) activity, the expression of Runx2 and VEGF, alizarin red S staining, and the quantification of osteocalcin and VEGF secretion in the periosteal-derived cells. The ALP activity increased rapidly up to day 14, followed by decrease in activity to day 35. Runx2 was expressed strongly at day 7, followed by decreased expression at day 14, and its expression was not observed thereafter. Both VEGF 165 and VEGF 121 were expressed strongly at day 35 and 42 of culture, particularly during the later stages of differentiation. Alizarin red S-positive nodules were first observed on day 14 and then increased in number during the entire culture period. Osteocalcin and VEGF were first detected in the culture medium on day 14, and their levels increased thereafter in a time-dependent manner. These results suggest that VEGF secretion from cultured human periosteal-derived cells increases along with mineralization process of the extracellular matrix. The level of VEGF secretion from periosteal-derived cells might depend on the extent of osteoblastic differentiation.

The biologic effects of safflower(Carthamus tinctorius $Linn\acute{e}$) extract and Dipsasi Radix extract on periodontal ligament cells and osteoblastic cells (홍화 추출물이 치주인대세포, 조골세포 활성도에 미치는 영향)

  • Rhyu, In-Chul;Lee, Yong-Moo;Ku, Young;Bae, Ki-Whan;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.27 no.4
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    • pp.867-882
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    • 1997
  • Safflower(Carthamus tinctorius $Linn\acute{e}$ has been traditionally used for the treatment of blood stasis, and Dipsasi Radix has been used as a drug for fracture in Chinese medicine. The purpose of present study was to examine the biologic effects of safflower extract and Disasi radix extracts on the periodontal. ligament cells and osteoblastic cells and on the wound healing of rat calvarial defect. The ethanolic extract of safflower blossom, safflower seed and Dipsasi Radix(125, 250, and 500 ${\mu}g/ml$) were prepared as test group, and PDGF-BB(lOng/ml) and unsafonifiable fraction of Zea Mays L.(125, 250, and 500 ${\mu}g/ml$) were employed as positive control. The effects of each agents on the growth and survival, ALPase activity, expression of PDGF-BB receptor, chemotactic response of PDL cell and ATCC human osteosarcoma MG63 cells in vitro were examined. The tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8mm defect in rat calvaria after oral administration of 3 different dosages groups : 0.02, 0.1 and 0.35g/kg, per day. It was also employed the same dosages of unsaponifiable fraction of Zea Mays L. as positive controls. Safflower blossom extract, safflower seed extract, and Dipsasi Radix extract stimulate the cellular activity of MG63 cells in concentration range of $125-500{\mu}g/ml$, and safflower bolssom extract and safflower seed extract stimulate also the cellular activity of periodontal ligament cells in concentration range of $250-500{\mu}g/ml$. In activity of ALPase, $250-500{\mu}g/ml$ of safflower blossom extracts showed significant stimulating effects on MG63 cells, and the same concentration range of safflower seed extracts showed significant effect on periodontal ligament cells. In the recovery on PDGF-BB receptor expression which was depressed by $IL-1{\beta}$, $125-250{\mu}g/ml$ of safflower blossom extracts and $250-500{\mu}g/ml$ of safflower seed extracts showed significant increasing effect on MG63 cells, and $500{\mu}g/ml$ of safflower blossom extract and $250-500{\mu}g/ml$ of safflower seed extracts showed significant effect on periodontal ligament cells. In chemotactic response, among all tested group, safflower seed extracts only were chemotactic to MG63 cells and periodontal ligament cells in concentration range of $125-500{\mu}g/ml$. Also in the view of bone regeneration in rat calvarial defect model, the only group that was orally administrated 0.35g/kg, day of safflower seed extract showed significant new bone formation. These results suggested that safflower extracts might have a potential possibilities as an useful drug for adjunct to treatment for regeneration of periodontal defect.

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Effect of $CO_2$ Enrichment on the Differentiation of Multi-shoots and Saponin contents in Tissue culture of Korean ginseng (Panax ginseng C. A. Meyer) (인삼(人蔘) 조직배양(組織培養)에서 $CO_2$처리(處理)가 multi-shoot 분화(分化) 및 사포닌 함량(含量)에 미치는 영향(影響))

  • Chung, Chan-Moon;Bae, Kil-Kwan
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.4
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    • pp.296-302
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    • 1999
  • This experiment was conducted to study the effect of $CO_2$(0, 2, 500, 5, 000, 10, 000ppm) enrichment by enabling ventilation on micropropagation of multi-shoot and on the saponin contents in vitro in Korean ginseng (Panax ginseng C. A. Meyer). Embryo was cultured in Murashige and Skoog medium added 3mg/ l of Indolbutyric acid, Benzyladenin and Gibberellic acid $(GA_3)$, respectively. $CO_2$, enrichment had little effects on the number of adventitious buds and shoots originated from adventitious buds. The ratio of differentiated shoots to adventitious buds were about 50% in $CO_2$, enrichment treatment. The shoots originated from adventitious bud showed more rapid growth and had larger leaf area than the shoots originated from the leaf primordia did. The number of shoot primordia was the highest in 2, 500ppm of $CO_2$ enrichment treatment. On the contrary, 10,000ppm of $CO_2$, enrichment made smaller the number of shoot primordia and ratio of shoots to shoot primordia. The range of shoots differentiated was from shoot primordia were 15. 4 to 23. 9. The rate of dry weight of cultured shoots showed lowest (7. 5%) in control and highest (8. 59%) in 2, 500ppm of $CO_2$, enrichment. Rate of in vitro flower in control was 7.6% and that in 2500ppm of $CO_2$ was about twice (15.7-16.3%) as much as in control. Flower number per a embryo cultured was about 1.2-1.3. In the multi-shoots with callus enriched by 2, 500ppm of $CO_2$, the contents of crude saponin and ginsenosides in multi-shoots alone were higher than in multi-shoots with callus. The characteristics of ginsenosides in multi-shoots were especially the higher content of ginsenoside Rd, Re, and $Rg_1$.

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