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      • Improvement of agricultural traits of SMV-resistant and drought-tolerant soybean transgenic plants

        Hye Jeong Kim,Jung Hun Pak,Mi-Jin Kim,Ha Neui Hong,Quyen Nguyen Thi,Yeong Hoon Lee,Ki Hyun Ryu,Kook-Hyung Kim,Soo Young Kim,Young Soo Chung 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07

        Korean soybean variety Kwangan was transformed with coat protein (CP), helper component-proteinase (HC-Pro), and ABRE binding factor 3 (ABF3) genes using highly efficient soybean transformation system. Among these genes, CP and HC-Pro were transformed using RNAi technology. Transgenic plants with CP were confirmed for gene introduction and their expression using PCR, real-time PCR, RT-PCR, Southern blot, and Northern blot. To investigate the response of viral infection with CP, T1 plants were inoculated with SMV-infected leaves and confirmed the existence of mosaic symptom in both leaves and seeds. Two transgenic lines with CP were highly resistant to SMV with clear leaves and seeds while SMV-susceptible lines showed mosaic symptom with seed mottling. The transcript levels of T1 plants with CP were also determined by northern blot, suggesting that SMV-resistant T1 plants did not show viral RNA expression whereas SMV-susceptible T1 plants showed viral RNA expression. Currently, the response of viral infection with HC-Pro is investigating to produce SMV-resistant soybean transgenic plants, and the physiological experiment with ABF3 is also carrying out to produce drought-tolerant soybean transgenic plants.

      • KCI등재

        Effect of Electroacupuncture on ERK Activation in Carrageenan-induced Inflammatory Pain Model

        Kim, Ji-Hwan,Lee, Si-Hyoung,Kim, Ha-Neui,Kim, Yu-Ri,Lee, Yong-Tae,Choi, Byung-Tae The Physiological Society of Korean Medicine and T 2010 동의생리병리학회지 Vol.24 No.5

        The present study found that EA pre-treatment effectively attenuated both spinal ERK activation and hyperalgesia against carrageenan-induced inflammation, whereas EA co-treatment with carrageenan injection unexpectedly elevated ERK activation in a synergistic manner and virtually had no analgesic effect. Therefore, we have concluded that the molecular mechanism of EA anagesia may be related to the inhibition of spinal ERK activation. Further experiments are required to find the intermediate candidates which transmits the pain-alleviating signals on the way of inhibiting ERK activation by EA.

      • KCI등재

        A single fraction from Uncaria sinensis exerts neuroprotective effects against glutamate-induced neurotoxicity in primary cultured cortical neurons

        Ha Neui Kim,Ji Yeon Jang,Byung Tae Choi 대한해부학회 2015 Anatomy & Cell Biology Vol.48 No.2

        We identified a neuroprotective single fraction among 62 ones of hexane extract from Uncaria sinensis (JGH43IA) and investigated its effects and mechanisms in primary cortical neurons. Pretreatment with JGH43IA showed a significantly increase cell viability in a dose-dependent manner with a decrease in the lactate dehydrogenase release. When we performed morphological assay and flow cytometry to determination of the type of cell death, pretreatment with JGH43IA showed a significant reduction of glutamate-induced apoptotic cell death. Then we explored the downstream signaling pathways of N-methyl-D-aspartate receptor (NMDAR) with calpain activation to elucidate possible pathways of neuroprotection by JGH43IA. Pretreatment with JGH43IA exhibited a significant attenuation of NMDAR GluN2B subunit activation and a decrease in active form of calpain 1 leading to subsequent cleavage of striatal-enriched protein tyrosine phosphatase (STEP). In addition, pretreatment with JGH43IA showed a marked increase of cAMP responsive element binding protein. These results suggest that JGH43IA may have neuroprotective effects through down-regulation of NMDAR GluN2B subunit and calpain 1 activation, and subsequent alleviation of STEP cleavage. This single fraction from U. sinensis might be a useful therapeutic agent for brain disorder associated with glutamate injury.

      • Effects of Electroacupuncture on <i>N</i>-Methyl-<i>D</i>-aspartate Receptor-Related Signaling Pathway in the Spinal Cord of Normal Rats

        Kim, Ha-Neui,Kim, Yu-Ri,Jang, Ji-Yeon,Shin, Hwa-Kyoung,Choi, Byung-Tae Hindawi Publishing Corporation 2012 Evidence-based Complementary and Alternative Medic Vol.2012 No.-

        <P>This study examined the influence of the N-methyl-D-aspartate receptor (NMDAR) on the modulation of related spinal signaling after electroacupuncture (EA) treatment in normal rats. Bilateral 2 Hz EA stimulations (1-2-3.0 mA) were delivered at acupoints corresponding to Zusanli (ST36) and Sanyinjiao (SP6) in men for 30 min. Thermal sensitization was strongly inhibited by EA, but this analgesia was reduced by preintrathecal injection of the NMDAR antagonist, MK801. Phosphorylation of the NMDAR NR2B subunit, cAMP response element-binding protein (CREB), and especially phosphatidylinositol 3-kinase (PI3K) were significantly induced by EA. However, these marked phosphorylations were not observed in MK801-pretreated rats. EA analgesia was reduced by preintrathecal injection with the calcium chelators Quin2 and TMB8, similar to the results evident using MK801. Phosphorylation of PI3K and CREB induced by EA was also inhibited by TMB8. Calcium influx by NMDAR activation may play an important role in EA analgesia of normal rats through the modulation of the phosphorylation of spinal PI3K and CREB.</P>

      • Production of soybean transgenic plants with Bt crystal protein genes

        Ha Neui Hong,Jung Hun Pak,Mi-Jin Kim,Hye Jeong Kim,Quyen Nguyen Thi,Yeon Ho Je,Young Soo Chung 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07

        Bacillus thuringiensis (Bt) crystal protein genes encode insecticidal δ-endotoxins that are widely used for the development of insect-resistant crops. Common soybean is a crop of economic and nutritious importance in many parts of the world. Korea soybean variety Kwangan was transformed with Bacillus thuringiensis (Bt) crystal protein genes. These genes were transformed into Kwangan using highly efficient soybean transformation system. Transgenic plants with Bt crystal protein genes were confirmed for gene introduction and their expression using PCR, real-time PCR and RT-PCR. Currently, the confirmation of stable gene introduction with Bt genes is also performing by southern blot analysis and physiology test and agronomic characters are investigating.

      • SCISCIESCOPUS

        Comparative analysis of the beneficial effects of treadmill training and electroacupuncture in a rat model of neonatal hypoxia-ischemia

        Kim, Ha Neui,Pak, Malk Eun,Shin, Myung Jun,Kim, Soo Yeon,Shin, Yong Beom,Yun, Young Ju,Shin, Hwa Kyoung,Choi, Byung Tae D.A. Spandidos 2017 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.39 No.6

        <P>In the present study, we investigated whether treadmill training and electroacupuncture (EA) have autonomous or synergistic beneficial effects on deficits caused by neonatal hypoxia-ischemia in Sprague-Dawley rats. For this purpose, rats subjected to hypoxia-ischemia underwent treadmill training and EA stimulation from 4 to 8 weeks of age. Conventional EA (CEA) and scalp EA (SEA) were delivered by electrical stimulation (2 Hz, 1 mA) at traditional acupoints and at the scalp to the primary motor area, respectively. In the behavioral examination, markedly improved performances in the rotarod test were observed in the rats that underwent treadmill exercise, and in the rats that underwent treadmill exercise and CEA compared to the untreated rats subjected to hypoxia-ischemia. An improvement was also observed in the passive avoidance test in the rats that underwent treadmill training and EA. As shown by western blot analysis, the expression levels of neuronal nuclei (NeuN), 2′,3′-cyclic-nucleotide 3′-phosphodiesterase and myelin basic protein (MBP) exhibited a significant decrease in the contralateral subventricular zone (SVZ) of the rats subjected to hypoxia-ischemia compared to the controls; however, these expression levels increased following treadmill exercise and EA stimulation. As shown by immunohistochemical analyses, the thickness of the corpus callosum and the integrated optical density (IOD) of MBP were significantly increased in the rats subjected to treadmill exercise and EA compared to the untreated rats subjected to hypoxiaa-ischemia. The synergistic effects of treadmill training and EA were also observed in the protein levels and IOD of MBP. A marked increase in the number of bromodeoxyuridine (BrdU)- and BrdU/NeuN-positive cells in the contralateral SVZ was also observed in the rats that underwent treadmill training and EA; the number of BrdU-positive cells was synergistically affected by treadmill training and EA. These results suggest that treadmill training and EA stimulation contribute to the enhancement of behavioral recovery following hypoxia-ischemia via the upregulation of myelin components and neurogenesis. Thus, treatment with EA stimulation, as well as treadmill training offers another treatment option to promote functional recovery in cerebral palsy.</P>

      • Beneficial effects of Jiawei Shenqi-wan and treadmill training on deficits associated with neonatal hypoxic-ischemia in rats

        Kim, Ha Neui,Pak, Malk Eun,Shin, Myung Jun,Kim, Soo Yeon,Shin, Yong Beom,Yun, Young Ju,Shin, Hwa Kyoung,Choi, Byung Tae D.A. Spandidos 2017 Experimental and therapeutic medicine Vol.13 No.5

        <P>Jiawei Shenqi-wan (JSQW), which comprises Shenqi-wan and two additional medicinal herbs, has been widely used for the treatment of various growth impairments, including cerebral palsy. In the present study, JSQW was administered to hypoxic-ischemic Sprague-Dawley rats that underwent treadmill training from 4–7 weeks of age to examine the beneficial effects of combined JSQW and treadmill therapy. Behavioral examinations were performed and a significant improvement in cylinder test performance was observed in rats treated with treadmill training compared with hypoxic-ischemia rats (P<0.05), as well as a significant improvement in passive avoidance test performance for rats treated with JSQW (P<0.05). The thickness of the corpus callosum and the integrated optical density (IOD) of myelin basic protein (MBP) were significantly increased by treatment with treadmill therapy alone (P<0.01 and P<0.001, respectively) and treatment with both JSQW and treadmill significantly increased the IOD of MBP compared with hypoxic-ischemia rats (P<0.001). Western blot analysis revealed that the expression of neuronal nuclei (NeuN) and doublecortin (Dcx) significantly decreased (P<0.001 and P<0.05, respectively) and MBP expression markedly decreased in the ipsilateral subventricular zone of hypoxic-ischemic rats compared with the control group; however, the expression of NeuN was significantly recovered by treatment with both JSQW and treadmill training (P<0.05). Furthermore, Dcx expression was significantly recovered by treatment with JSQW (P<0.05), and MBP expression was significantly restored by treatment with treadmill training (P<0.01). In the immunohistochemical analyses, a significant increase in the number of bromodeoxyuridine (BrdU) positive cells in this region was observed in treadmill-treated rats (P<0.05), whereas significant increases in the number of Brdu/Dcx or NeuN or glial fibrillary acidic protein double-positive cells were observed only in the group co-treated with JSQW and treadmill (P<0.01, P<0.05 and P<0.001, respectively). These results suggest that JSQW and treadmill training may contribute to behavior recovery following hypoxic-ischemia, and JSQW treatment was particularly effective in promoting memory function via enhancing the differentiation of neuronal progenitor cells. The results of the present study therefore suggest that JSQW may provide an additional treatment option for functional recovery with treadmill training in cerebral palsy.</P>

      • Electroacupuncture Confers Antinociceptive Effects via Inhibition of Glutamate Transporter Downregulation in Complete Freund's Adjuvant-Injected Rats

        Kim, Ha-Neui,Kim, Yu-Ri,Jang, Ji-Yeon,Shin, Hwa-Kyoung,Choi, Byung-Tae Hindawi Publishing Corporation 2012 Evidence-based Complementary and Alternative Medic Vol.2012 No.-

        <P> When we evaluated changes of glial fibrillary acidic protein (GFAP) and two glutamate transporter (GTs) by immunohistochemistry, expression of GFAP showed a significant increase in complete Freund's adjuvant (CFA)-injected rats; however, this expression was strongly inhibited by electroacupuncture (EA) stimulation. Robust downregulation of glutamate-aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1) was observed in CFA-injected rats; however, EA stimulation resulted in recovery of this expression. Double-labeling staining showed co-localization of a large proportion of GLAST or GLT-1 with GFAP. Using Western blot, we confirmed protein expression of two GTs, but no differences in the mRNA content of these GTs were observed. Because EA treatment resulted in strong inhibition of CFA-induced proteasome activities, we examined the question of whether thermal sensitivities and GTs expression could be regulated by proteasome inhibitor MG132. CFA-injected rats co-treated with EA and MG132 showed a significantly longer thermal sensitivity, compared with CFA-injected rats with or without MG132. Both EA and MG132 blocked CFA-induced GLAST and GLT-1 downregulation within the spinal cord. These results provide evidence for involvement of GLAST and GLT-1 in response to activation of spinal astrocytes in an EA antinociceptive effect. Antinociceptive effect of EA may be induced via proteasome-mediated regulation of spinal GTs.</P>

      • alpha-Lipoic acid inhibits inflammatory bone resorption by suppressing prostaglandin E2 synthesis.

        Ha, Hyunil,Lee, Jong-Ho,Kim, Ha-Neui,Kim, Hyun-Man,Kwak, Han Bok,Lee, Seungbok,Kim, Hong-Hee,Lee, Zang Hee Williams Wilkins 2006 JOURNAL OF IMMUNOLOGY Vol.176 No.1

        <P>alpha-Lipoic acid (LA) has been intensely investigated as a therapeutic agent for several pathological conditions, including diabetic polyneuropathy. In the present study, we examined the effects of LA on osteoclastic bone loss associated with inflammation. LA significantly inhibited IL-1-induced osteoclast formation in cocultures of mouse osteoblasts and bone marrow cells, but LA had only a marginal effect on osteoclastogenesis from bone marrow macrophages induced by receptor activator of NF-kappaB ligand (RANKL). LA inhibited both the sustained up-regulation of RANKL expression and the production of PGE2 induced by IL-1 in osteoblasts. In addition, treatment with either prostaglandin E2 (PGE2) or RANKL rescued IL-1-induced osteoclast formation inhibited by LA or NS398, a specific cyclooxygenase-2 (COX-2) inhibitor, in cocultures. LA blocked IL-1-induced PGE2 production even in the presence of arachidonic acid, without affecting the expression of COX-2 and membrane-bound PGE2 synthase. Dihydrolipoic acid (the reduced form of LA), but not LA, attenuated recombinant COX-2 activity in vitro. LA also inhibited osteoclast formation and bone loss induced by IL-1 and LPS in mice. Our results suggest that the reduced form of LA inhibits COX-2 activity, PGE2 production, and sustained RANKL expression, thereby inhibiting osteoclast formation and bone loss in inflammatory conditions.</P>

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