• Journal of Korean Society of Forest Science
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• v.99 no.1
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• pp.85-95
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• 2010

The purpose of this study was to quantitatively measure both growth performances and nutrient responses of Fraxinus rhynchophylla, Fraxinus mandshurica, Pinus koraiensis, and Abies holophylla seedlings, which are commercially planted in Korea, to nitrogen, phosphorus, and potassium fertilization. We used Dickson's quality index (QI) to compare growth performances and vector diagnosis to interpret nutrient status. Nitrogen fertilization increased more height and root collar diameter growth in F. rhynchophylla and F. mandshurica relative to no fertilization treatment. The QI of F. rhynchophylla and F. mandshurica was the highest on N treatment, but there were no significant differences between treatments for P. koraiensis and A. holophylla. Nitrogen fertilization increased total dry weight by 43, 41, 26, -9% for F. rhynchophylla, F. mandshurica, P. koraiensis and A. holophylla, respectively. In F. rhynchophylla, N fertilization increased N contents with similar N concentrations ("sufficiency"), decreased both P concentrations and P contents ("antagonism"), and decreased K contents with similar K concentrations ("toxic accumulation"). In P. koraiensis, N fertilization decreased N, P, and K concentrations because of more dry weight increases compared to uptaken contents ("dilution"), but N fertilization decreased N, P, and K contents with similar N, P, and K concentrations ("toxic accumulation"). In the light of quality index and vector diagnosis, F. rhynchophylla and F. mandshurica seedlings treated with N fertilization would have high field performance.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.3
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• pp.199-218
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• 2005

Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.