• Microbiology and Biotechnology Letters
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• v.34 no.2
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• pp.129-135
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• 2006

A gene encoding for a putative microsomal epoxide hydrolase (mEH) of a zebrafish, Danio rerio, was cloned and characterized. The putative mEH protein of D. rerio exhibited sequence similarity with mammalian mEH and some other bacterial EHs. A structural model for the putative mEH was constructed using homology modeling based on the crystallographic templates, 1 qo7 and 1 ehy. The catalytic triad consisting of $Asp^{233}$, $Glu^{413}$, and $His^{440}$ was identified, and the characteristic features such as two tyrosine residues and oxyanion hole were found to be highly conserved. Based on bioinformatic analysis together with EH activity assay, the putative protein was annotated as mEH of D. rerio. Enantiopure styrene oxide with enantiopurity of 99%ee and yield of 33.5% was obtained from racemic styrene oxide by the enantioselective hydrolysis activity of recombinant mEH of D. rerio for 45 min.

• Journal of Acupuncture Research
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• v.33 no.3
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• pp.1-16
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• 2016

Objectives : The objective of this study was to investigate the effects of Gentianae Macrophyllae Radix pharmacopuncture (GP) at Hwando (GB30) in neuropathic pain induced rats. Methods : Neuropathic pain in rats was induced by tibial and sural nerve transection. The rat subjects were divided into 6 groups : normal (Nor, n = 5), control (Con, n = 5), neuropathic pain- induced injected at GB30 with 1 mg/kg GP (GP-A, n = 5), 5 mg/kg GP (GP-B, n = 5) and 20 mg/kg GP (GP-C, n = 5), and neuropathic pain-induced injected with 1mg/kg Tramadol (Tramadol, n＝5). Injections were administered 2 times a week for a total of 5 treatments. After each treatment plantar withdrawal response was measured and after all 5 treatments were completed c-fos, Bax, Bcl-2, mGlu5 and leukocytes in the blood were analyzed. Results : 1. Groups GP-A, GP-B and GP-C showed a meaningful decrease in the withdrawal response of mechanical allodynia compared to the control group. 2. Groups GP-A, GP-B and GP-C showed a meaningful decrease in the expression of c-fos compared to the control group. 3. Groups GP-A and GP-C showed a meaningful increase in the expression of mGluR5 compared to the control group. 4. Groups GP-A, GP-B and GP-C showed a meaningful decrease in Bax/Bcl-2 ratio compared to the control group. Conclusion : These results suggest that Gentianae Macrophyllae Radix pharmacopuncture at Hwando (GB30) could decrease mechanical allodynia and could have analgesic and neuroprotective effects on the model of neuropathic pain.

• Korean Journal of Acupuncture
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• v.25 no.1
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• pp.229-246
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• 2008

Objectives : Acupoints of LR1(Dadun)${\cdot}$HT9(Shaochong), KI10(Yingu)${\cdot}$HT3(Shaohai) are used as controlling the diseases from heart problems. Especially, when they are used, tonification or sedation techniques method is taken as a controlling the medication for an early stroke in the Korean medicine. So the aim of this study is to investigate the anti-apoptotic and neuroprotective effects of Acupuncture techniques of tonification or sedation at LR1${\cdot}$HT9, KI10${\cdot}$HT3 on the focal ischemia induced by intraluminal filament insertion in rats. Materials and Methods : The focal ischemia was induced by intraluminal filament insertion into middle cerebral artery. The animals were divided into five groups (n=7 in each group) : Control group, no therapy group after ischemia induced; AT-1; acupuncture therapy group at LR1${\cdot}$HT9, KI10${\cdot}$HT3 after ischemia-induced. AT-2; acupuncture therapy at right LR1, HT9(forward direction), KI10${\cdot}$HT3(opposite direction) inserting to the direction of route of the each meridian. AT3; acupuncture therapy at right LR1, HT9(twirling forward with the thumb of right hand 9 times), KI10${\cdot}$HT3(twirling forward with the forefinger of right hand 6 times) AT-4; acupuncture therapy at right LR1, HT9(forward direction, twirling forward with the thumb of right hand 9 times), KI10${\cdot}$HT3(opposite direction, twirling forward with the forefinger of right hand 6 times) inserting to the direction of route of the each meridian. The anti-apoptotic and neuroprotective effects of Acupuncture techniques of tonification or sedation at LR1, HT9, KI10${\cdot}$HT3 were observed by Bax, Bcl-2, mGluR5, Cytochrome c, Cresyl violet and ChAT-stain. Results : The intensity of Bax and Bax/Bcl-2 ratio were increased in ACU-2 and ACU-4 group but decreased in ACU-3 group. The intensity of mGluR5 was increased in ACU-1, ACU-2, ACU-3 and ACU-4 group. The intensity of cytochrome c was decreased in ACU-3 and ACU-4 group. The density of neurons stained by Cresyl violet and ChAT were increased in ACU-1, ACU-3 and ACU-4 group. Conclusions : Our study suggests that acupuncture therapy of tonification at LR1, HT9 by twirling forward with the thumb of right hand 9times, sedation at KI10${\cdot}$HT3 by twirling forward with the forefinger of right hand 6 times after perpendicularly inserting needle shows anti-apoptotic and neuroprotective effects on cholinergic neuron in focal cerebral ischemia of the stroke in rats.

• Journal of Photoscience
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• v.9 no.2
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• pp.305-307
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• 2002

Spectroscopic titration of D 193N and D 193E mutants of pharaonis phoborhodopsin (ppR) were performed to evaluate the pK$_{a}$ of the Schiff base Asp 193 corresponds to Glu204 of bacteriorhodopsin (bR). The pK$_{a}$ of the Schiff base (SBH$^{+}$) of D193N was 10.1~10.0 (at XH$^{+}$) and 11.4~11.6 (at X) depending on the protonation state of a certain residue (designated by X) and independent on CI$^{[-10]}$ , while those of the wild-type and D193E were> 12. pK$_{a}$ of XH$^{+}$ were; 11.8~11.2 at the state of SB, 10.5 at SBH$^{+}$ state in the presence of CI$^{[-10]}$ , and 9.6 at SBH$^{+}$ without CI$^{[-10]}$ These imply the presence of a long-range interaction in the extracellular channel.r channel.

• Mycobiology
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• v.43 no.3
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• pp.311-318
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• 2015

Culture filtrates of six different edible mushroom species were screened for antimicrobial activity against tomato wilt bacteria Ralstonia solanacearum B3. Hericium erinaceus, Lentinula edodes (Sanjo 701), Grifola frondosa, and Hypsizygus marmoreus showed antibacterial activity against the bacteria. Water, n-butanol, and ethyl acetate extracts of spent mushroom substrate (SMS) of H. erinaceus exhibited high antibacterial activity against different phytopathogenic bacteria: Pectobacterium carotovorum subsp. carotovorum, Agrobacterium tumefaciens, R. solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. axonopodis pv. citiri, and X. axonopodis pv. glycine. Quantitative real-time PCR revealed that water extracts of SMS (WESMS) of H. erinaceus induced expressions of plant defense genes encoding ${\beta}$-1,3-glucanase (GluA) and pathogenesis-related protein-1a (PR-1a), associated with systemic acquired resistance. Furthermore, WESMS also suppressed tomato wilt disease caused by R. solanacearum by 85% in seedlings and promoted growth (height, leaf number, and fresh weight of the root and shoot) of tomato plants. These findings suggest the WESMS of H. erinaceus has the potential to suppress bacterial wilt disease of tomato through multiple effects including antibacterial activity, plant growth promotion, and defense gene induction.

• Journal of Oriental Neuropsychiatry
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• v.22 no.4
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• pp.201-218
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• 2011

Objectives : The aim of this study was to investigate the effects of Guibi-Tang(GBT) on focal brain ischemic injury induced by intraluminal filament insertion in rats. Methods : The ischemia was induced by intraluminal filament insertion into middle cerebral artery. Sprague-Dawley rats were divided into five groups, normal group(n=8); control group was ischemia induced and no treatment(n=8); GBT 1X group was ischemia induced and administrated 42.2 mg/ml/kg of Guibi-Tang orally(n=8); GBT 3X group was ischemia induced and administrated 126.6 mg/ml/kg of Guibi-Tang orally(n=8); GBT 6X group was ischemia induced and administrated 253.2 mg/ml/kg of Guibi-Tang orally(n=8) for 21 days. mGluR5, Bax, Bcl-2 and cytochrome C were investigated to observe the effects of Guibi-Tang on apoptosis. The effects of Guibi-Tang on neuroprotective/apoptotic agents in cresyl violet, choline acetyltranferase(ChAT) with ischemic injury were investigated. Results : The intensity of mGluR5 mRNA in the hippocampal CA1 was increased in normal and GBT(Guibi-Tang) 1X groups compared with the control group. The intensity of Bax mRNA in the hippocampal CA1 was decreased in normal and GBT 1X groups compared with the control group. However it was increased unexpectedly in GBT 3X group. The intensity of Bcl-2 mRNA in the hippocampal CA1 was increased in normal and GBT 1X groups compared with the control group. The intensity of Bax/Bcl-2 ratio in the hippocampal CA1 was decreased in normal and GBT 1X groups compared with the control group. The intensity of cytochrome C protein in the hippocampal CA1 was decreased in normal, GBT 1X and GBT 6X groups compared with the control group. The density of neurons stained by cresyl violet and ChAT was increased in normal and GBT 1X groups compared with the control group. Conclusions : These results suggest that Guibi-Tang may have protective effect on vascular dementia.

• Fisheries and Aquatic Sciences
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• v.14 no.3
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• pp.174-178
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• 2011

This study aimed to determine the degree of hydrolysis and angiotensin-I-converting enzyme (ACE)-inhibitory activity of Giant Jellyfish Nemopilema nomurai (jellyfish) hydrolysates. The degree of hydrolysis using six proteolytic enzymes (Alcalase, Flavozyme, Neutrase, papain, Protamex, and trypsin) ranged from 13.1-36.8% and the inhibitory activities from 20.46-79.58%. Using papain hydrolysate, we newly isolated and characterized ACE-inhibitory peptides with a molecular weight of 3,000-5,000 Da that originated from jellyfish collagen. The purified peptide (FII-b) was predicted to be produced from an alpha-2 fragment of the type IV collagen of jellyfish. The N-terminal sequence of FII-b was Asp-Pro-Gly-Leu-Glu-Gly-Ala-His-Gly- and showed 87% identity to the collagen type IV alpha-2 fragment of Rattus norvegicus and a predicted protein from Nematostella vectensis, indicating that the ACE-inhibitory peptide originated from the collagen hydrolysate and had an $IC_{50}$ value of 3.8 ${\mu}g$/mL. The primary structure of the fragment is now being studied; this peptide represents an interesting new type of ACE inhibitor and will provide knowledge of the potential applications of jellyfish components as therapies for hypertension.

• Journal of Acupuncture Research
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• v.33 no.4
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• pp.49-63
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• 2016

Objectives : This research was performed to investigate the effects of Jodeungsan pharmacopuncture(PA-J) of focal brain ischemia induced by middle cerebral artery occlusion(MCAO) in rats. Methods : The subjects were divided into 4 groups : control, acupuncture, pharmacopuncture PA-J1(11.43 mg / 250 g / $40{\mu}{\ell}$) and pharmacopuncture PA-J2(2.29 mg / 250 g / $40{\mu}{\ell}$). The focal brain ischemia was induced by intraluminal filament insertion into the middle cerebral artery. After 3 days of MCAO, Jodeungsan pharmacopuncture treatment was performed on the GB20, and the day after being treated with pharmacopuncture, the Morris water maze test was carried out on the assigned group. The series of processes were administered 6 times. Thereafter mGluR5, density of neuronal cell and ChAT were measured. Results : The results were as follows. 1. The distance to target significantly decreased in the 2nd trial of the Acu group on the water maze test for short-term memory. 2. The distance to target significantly decreased in the 4th trial of the PA-J2 group on the water maze test for long-term memory. 3. The intensity of mGluR5 significantly increased in the PA-J1 group compared with the control group. 4. The neuroprotective effect on the hippocampal CA1 significantly increased in the PA-J1 and PA-J2 groups compared with the control group. 5. The density of ChAT in the hippocampal CA1 significantly increased in the PA-J1 and PA-J2 groups compared with the control group. Conclusion : These results suggest that Jodeungsan pharmacopuncture may improve memory and cognitive impairment and also have neuroprotective effects on focal brain ischemia.

• Food Science of Animal Resources
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• v.37 no.3
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• pp.402-409
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• 2017

A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

• Journal of Acupuncture Research
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• v.31 no.4
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• pp.57-70
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• 2014

Objectives : The purpose of this study is to examine if Clematidis Radix(CR) pharmacopuncture may be effective to the neuropathic pain(mechanical allodynia) in a rat model of neuropathic pain. Methods : To produce the model of neuropathic, L5 spinal nerve was ligated by 6-0 silk thread. After neuropathic surgery, the author examined if the animals exhibited the plantar withdrawal response of allodynia. The plantar withdrawal response was assessed by dynamic plantar aesthesiometer three days after the neuropathic surgery, CR pharmacopuncture was injected at $BL_{23}$ 1time/week for 6 weeks. After that, the author examined the plantar withdrawal response of rats' leg by dynamic plantar aesthesiometer. And also the author examined mGluR5, Bax, Bcl-2, Bax/Bcl-2 ratio in spinal cord, and c-Fos. Also the author observed the change of aspartate aminotransferase(AST), alanine aminotransferase(ALT) count in the blood serum of neuropathic rats. Results : 1. The withdrawal response of allodynia decreased in the PT3, PT4 group as compared with control group. 2. The mGluR5 increased in the PT1, PT2, PT3, and PT4 group. 3. The Bax and Bax/Bcl-2 ratio decreased in the PT4 group. 4. The c-Fos increased in the PT1 group, and decreased in the PT4 group. 5. The changes of AST in blood serum decreased in the every group excluded control group and the changes of ALT in blood serum isn't shown the signigicant change. Conclusions : These results are suggested that CR pharmacopuncture at BL23 decreased mechanical allodynia, can act on anti-apoptotic, neuroprotective effects and liver fuction in the model of neuropathic pain.