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Studies of the Effects of Acupuncture Stimulation at Huatuo Jiaji(EX B2) Points on Axonal Regeneration of Injured Sciatic Nerve in the Rats (화타협척혈 침자극에 의한 손상 말초신경의 재생효과에 관한 연구)

  • Kim, Dae-Feel;Park, Young-Hoi;Keum, Dong-Ho
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.4
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    • pp.39-61
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    • 2008
  • Objectives : The present study was performed to investigate whether acupuncture stimulation in the rats affected regeneration properties of the injured sciatic nerve. A differential effect of acupuncture stimulation on the one point near the spinal nerve root controlling sciatic nerve activity and the other point in the peripheral area subordinated by injured nerve was compared. Materials and Methods: Rat sciatic nerves were injured by crush, and the effects on axonal regeneration on injured sciatic nerves were evaluated by acupuncture stimulation at two different regions. In proximal acupuncture stimulation group, acupuncture stimulation was performed on Huatuo Jiaji(EX B2) points located from L5 to S1 vertebral levels to stimulate the nearest spinal nerve root that innervates sciatic nerves. In distal acupuncture stimulation group, acupuncture stimulation was performed on Zusanli(ST 36) and Weizhong(BL 40) points to stimulate at peripheral area dominated by injured sciatic nerves. Acupuncture stimulation was given every other days for 1 or 2 weeks. Sciatic nerve tissues collected from acupuncture stimulation experimental groups, injury control group, and intact animal group were used for protein analysis by Western blotting or Hoechst nuclear staining. To determine axonal regeneration, Dil fluorescence dye was injected into the sciatic nerve 0.5 cm distal to the injury site in individual animal groups and Dil-labeled cells by retrograde tracing were measured in the DRG at lumbar 5 or in the spinal cord. DRG sensory neurons prepared from individual animal groups were used to measure the extent of neurite outgrowth and for immunofluorescence staining with anti-GAP-43 antibody. Results : Animal groups given proximal or distal acupuncture stimulation showed upregulation of GAP-43 and Cdc2 protein levels in the sciatic nerve at 7 days after injury. Cdk2 protein levels were strongly induced by nerve injury, but did not show changes by acupuncture stimulation. Phospho-Erk1/2 protein levels were elevated by acupuncture stimulation above those present in the injury control animals. These increase in regeneration-associated protein levels appeared to be related with increase cell proliferation in the injured sciatic nerves. Hoechst 33258 staining of sciatic nerve tissue to visualize nuclei of individual cells showed increased Schwann cell number in the distal portion of the injured nerve 7 and 14 days after injury and further increases by acupuncture stimulation particularly at the proximal position. Measurement of axonal regeneration by retrograde tracing showed significantly increased Dil-labeled cells in proximal acupuncture stimulation group compared to distal acupuncture stimulation group and injury control group. Finally, an evaluation of axonal regeneration by retrograde tracing showed increased number of Dil labeled cells in the DRG at lumbar 5 or in the ventral horn of the spinal cord at lower thoracic level at 7 days after nerve injury. Conclusions : The present data show that the proximal acupuncture stimulation at Huatuo Jiaji(EX B2) points governing injured sciatic nerves was more effective for axonal regeneration than the distal acupuncture stimulation. Further studies on functional recovery or associated molecular mechanisms should be critical for developing animal models and clinical applications.

Dismantling and Restoration of the Celadon Stool Treasure with an Openwork Ring Design (보물 청자 투각고리문 의자의 해체 및 복원)

  • KWON, Ohyoung;LEE, Sunmyung;LEE, Jangjon;PARK, Younghwan
    • Korean Journal of Heritage: History & Science
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    • v.55 no.2
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    • pp.200-211
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    • 2022
  • The celadon stools with an openwork ring design which consist of four items as one collection were excavated from Gaeseong, Gyeonggi-do Province. The celadon stools were designated and managed as treasures due to their high arthistorical value in the form of demonstrating the excellence of celadon manufacturing techniques and the fanciful lifestyles during the Goryeo Dynasty. However, one of the items, which appeared to have been repaired and restored in the past, suffered a decline in aesthetic value due to the aging of the treatment materials and the lack of skill on the part of the conservator, raising the need for re-treatment as a result of structural instability. An examination of the conservation condition prior to conservation treatment found structural vulnerabilities because physical damage had been artificially inflicted throughout the area that was rendered defective at the time of manufacturing. The bonded surfaces for the cracked areas and detached fragments did not fit, and these areas and fragments had deteriorated because the adhesive trickled down onto the celadon surface or secondary contaminants, such as dust, were on the adhesive surface. The study identified the position, scope, and conditions of the bonded areas at the cracks UV rays and microscopy in order to investigate the condition of repair and restoration. By conducting Fourier-transform infrared spectroscopy(FT-IR) and portable x-ray fluorescence spectroscopy on the materials used for the former conservation treatment, the study confirmed the use of cellulose resins and epoxy resins as adhesives. Furthermore, the analysis revealed the addition of gypsum(CaSO4·2H2O) and bone meal(Ca10 (PO4)6(OH)2) to the adhesive to increase the bonding strength of some of the bonded areas that sustained force. Based on the results of the investigation, the conservation treatment for the artifact would focus on completely dismantling the existing bonded areas and then consolidating vulnerable areas through bonding and restoration. After removing and dismantling the prior adhesive used, the celadon stool was separated into 6 large fragments including the top and bottom, the curved legs, and some of the ring design. After dismantling, the remaining adhesive and contaminants were chemically and physically removed, and a steam cleaner was used to clean the fractured surfaces to increase the bonding efficacy of the re-bonding. The bonding of the artifact involved applying the adhesive differently depending on the bonding area and size. The cyanoacrylate resin Loctite 401 was used on the bonding area that held the positions of the fragments, while the acrylic resin Paraloid B-72 20%(in xylene) was treated on cross sections for reversibility in the areas that provided structural stability before bonding the fragments using the epoxy resin Epo-tek 301-2. For areas that would sustain force, as in the top and bottom, kaolin was added to Epo-tek 301-2 in order to reinforce the bonding strength. For the missing parts of the ring design where a continuous pattern could be assumed, a frame was made using SN-sheets, and the ring design was then modeled and restored by connecting the damaged cross section with Wood epos. Other restoration areas that occurred during bonding were treated by being filled with Wood epos for aesthetic and structural stabilization. Restored and filled areas were color-matched to avoid the feeling of disharmony from differences of texture in case of exhibitions in the future. The investigation and treatment process involving a variety of scientific technology was systematically documented so as to be utilized as basic data for the conservation and maintenance.

Multiplication of Infectious Flacherie and Densonucleosis Viruses in the Silkworm, Bombyx mori (가잠의 전염성 연화병 및 농핵병 바이러스 증식에 관한 연구)

  • 김근영;강석권
    • Journal of Sericultural and Entomological Science
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    • v.25 no.2
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    • pp.1-31
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    • 1984
  • Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

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