• Journal of Korean Medicine Rehabilitation
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• v.19 no.1
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• pp.23-38
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• 2009

Objectives : The purpose of this morphological studies was to investigate the relations between Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas of rats using peudorabies virus(PRV). Methods : We observed labeled neurons following the injection of PRV, Bartha strain, into the Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas of rats. After survival times of 4 days following the injection of PRV, the rats were perfused, and their spinal ganglia, spinal cord and brain stem were frozen sectioned($35{\mu}m$). These sections were strained by PRV immunohistchemical staining methods and observed with light microscope. Results : The results were as follows. 1. In the spinal ganglia, the overlap areas of PRV labeled neurons projecting to Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas were observed in T10-13 dorsal root ganglia. 2. In the spinal cord, the overlap areas of PRV labeled neurons projecting to Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas were lamina I, IV, V, VII, IX, X, intermediolateral nucleus(IML), intermediomedial nucleus(IMM) in thoracic segments. In lumbar segments, the overlap areas of PRV labeled neuron were lamina I, IV, V, VI, IX, X and IMM. In sacral segments, the overlap areas of PRV labeled neuron were lamina I, IV, V, VI, VII, IX, X. 3. In the brain, the overlap areas of PRV labeled neurons projecting to Sanyinjiao(Sp6), Yinlingquan(Sp9) and pancreas were area postrema, nucleus tractus solitarius, caudoventrolateral reticular nu., medullary reticular nu., lateral paragigantocellular nu., C3 adrenalin cells, gigantocellular nu., raphe pallidus nu., raphe obscurus nu., ambiguus nu., raphe magnus nu., pontine reticular formation, A5 cell group, subcoeruleus nu., locus coeruleus, Barringnton's nu., $K{\ddot{o}}lliker$-Fuse nu., dorsal raphe nu., Edinger-Westphal nu., central gray matter, perifornical nu., dorsomedial hypothalamic nu., arcuate nu., lateral hypothalamic nu., paraventricular hypothalamic nu., hindlimb area. Conclusions : In conclusion, these results suggest that the interrelationship of meridian(spleen meridian), acupoints(Sp6 and Sp9) and viscera(pancreas) may be related the central autonomic centers.

• Journal of Acupuncture Research
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• v.20 no.6
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• pp.89-102
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• 2003

Objective: The expression of CRF(corticotropin releasing factor), CRF-R(receptor) and CRF-BP(binding protein) in CNS neurons projecting to the stomach and ST36 using the pseudorabies virus in the rat was investigated. Methods: After survival times of 5 days following injection of PRV-Ba-Gal, The thirty rats were perfused, and their brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV-Ba-Gal histochemical staining method and(or) CRF, CRF-R and CRF-BP immunohistochemical method. The common expressed areas of the brain projecting to the stomach and zusanli(ST36) following injection of PRV-Ba-Gal were observed with light microscope. Results: 1) The dense accumulation of CRF-immunoreactive terminals is seen in the area postrema, n. tractus solitarius, external zone of the median eminence, with some immunoreactive CRF also present in the internal zone. 2) Aggregates of CRF-R immunoreactive perikarya are found in area postrema, n. tractus solitarius, lateral reticular n., gigantocellular reticular n., locus coeruleues, paraventricular n. of hypothalamus, median eminence, preoptic n., arcuate n. and hind limb area of cerebral cortex. 3) Aggregates of CRF-BP immunoreactive perikarya are found in area postrema, n. tractus solitarius, lateral reticular nucleus, gigantocellular reticular n., locus coeruleues, paraventricular n. of hypothalamus, median eminence and arcuate n.. Conclusions : These results suggest that PRV-Ba-Gal labeled areas projecting to stomach and ST36 may be related to the central autonomic pathways. A part of CNS neurons projecting to the stomach and ST36 were related to expression of CRF, CRF-R and CRF-BP related to the stress in central autonomic center.

• Molecules and Cells
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• v.38 no.9
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• pp.773-780
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• 2015

3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and $10{\mu}g$ 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 $LD_{50}$ PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% ($5{\mu}g$) and 47% ($10{\mu}g$). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.140-147
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• 2016

A process for manufacturing virally-safe porcine bone hydroxyapatite (HA) has been developed to serve as advanced xenograft material for dental applications. Porcine bone pieces were defatted with successive treatments of 30% hydrogen peroxide and 80% ethyl alcohol. The defatted porcine bone pieces were heat-treated in an oxygen atmosphere box furnace at $1,300^{\circ}C$ to remove collagen and organic compounds. The bone pieces were ground with a grinder and then the bone powder was sterilized by gamma irradiation. Morphological characteristics such as SEM (Scanning Electron Microscopy) and TEM (Transmission Electron Microscopy) images of the resulting porcine bone HA (THE Graft$^{(R)}$) were similar to those of a commercial bovine bone HA (Bio-Oss$^{(R)}$). In order to evaluate the efficacy of $1,300^{\circ}C$ heat treatment and gamma irradiation at a dose of 25 kGy for the inactivation of porcine viruses during the manufacture of porcine bone HA, a variety of experimental porcine viruses including transmissible gastroenteritis virus (TGEV), pseudorabies virus (PRV), porcine rotavirus (PRoV), and porcine parvovirus (PPV) were chosen. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the $1,300^{\circ}C$ heat treatment. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.81$$ for PRV, $${\geq_-}6.28$$ for PRoV, and $${\geq_-}5.21$$ for PPV. Gamma irradiation was also very effective at inactivating the viruses. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the gamma irradiation. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.87$$ for PRV, $${\geq_-}6.05$$ for PRoV, and $${\geq_-}4.89$$ for PPV. The cumulative log reduction factors achieved using the two different virus inactivation processes were $${\geq_-}9.30$$ for TGEV, $${\geq_-}11.68$$ for PRV, $${\geq_-}12.33$$ for PRoV, and $${\geq_-}10.10$$ for PPV. These results indicate that the manufacturing process for porcine bone HA from porcine-bone material has sufficient virus-reducing capacity to achieve a high margin of virus safety.

• The Journal of Korean Medicine
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• v.22 no.2
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• pp.31-40
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• 2001

This experimental studies was to investigate location of labeled neurons in CNS following injection of pseudorabies virus(PRV), Bartha strain, into the uterus and Sanyinjiao(Sp6) of rats. After survival times of 4-5 days following the injection of PRV, the rats were perfused, and their brain and spinal cord were frozen sectioned($30\mu\textrm{m}$). These sections were stained by PRV immunohistochemical staining methods, and observed with light microscope. The results were as follows: 1. In the spinal cord, overlap areas of PRV labeled neurons projecting to uterus and Sp6 were observed in lamina VII, IX and X areas of cervical segments. In thoracic segments, overlap areas were observed in lamina IV, VII, X and intermediolateral n.. In lumbar segments, overlap area of PRV labeled neurons were observed in lamina I, V-VII, IX, X and intermediolateral n.. In sacral segments, overlap areas of PRY labeled neurons were observed in lamina N, V, VII, X and sacral parasympathetic n.. 2. In the brain, overlap areas of PR V labeled neurons projecting to the uterus and Sp6 were observed in lateral paragigantocellular n., rostroventrolateral reticular n., raphe obscurus n., raphe pallidus n., raphe magnus n., locus coeruleus n., Barrington's n., A5 cell group, central gray n., paraventricular hypothalamic n. and arcuate n. This results suggest that overlap areas of PRV labeled neurons of the spinal cord projecting to the uterus and Sp6 might be the first-order neurons related to the viscera-somatic sensory and sympathetic preganglionic neurons. PRV labeled neurons of the brain may be the second and third-order neurons response to the movement of smooth muscle of uterus. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitoring the internal environment. These overlap areas of spinal cord and brainmay be related to autonomic centers related to regulation of uterus.

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.463-470
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• 2000

Until now peudorabies virus(PRV) has been used a neurotracer, because of it's properties of retrograde & anterograde transport. But it's anterograde transfort is not perfect, so we tested the applicability of the Bartha strain of PRV(PRV-Ba) isolated from South Korea as a neurotracer in the visual system. We performed immunohistochemical study of the rabbit brain after intravitreal injection of the PRV-Ba. After given survival time(24, 48, 72, 96, 120, 144hrs), the brain was removed and processed immunohistochemical stain for PRV-Ba. The strong PRV immunoreactivity(PRV-ir) were almost oberserved contralaterally in oculomotor neurons, fro example Edinger-Westphal nucleus, trigerminal nucleus of pons and peritrigerminal zone but locus of innervating sensitive neurons. The latter were weak positive and selective. PRV-Ba immunoreactive neurons were stained strongly in nucleus compared to cytoplasm. This study suggests that PRV-Ba isolated from South Korea is also a useful neurotracer in the motor innervated system like other PRV-strain.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.4
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• pp.805-817
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• 2009

This study was to investigate central localization of neurons projecting to the urinary bladder and urinary bladder-related acupoints(Wijung, $BL_{40}$) and neurons of immunoreactive to hormones and hormone receptors regulating urinary bladder function by using peudorabies virus(PRV). In this experiment, Bartha's strain of pseudorabies virus was used in rats to trace central localization of urinary bladder-related neurons and urinary bladder-related acupoints($BL_{40}$) which can regulate urinary system. PRV was injected into the urinary bladder and acupoints($BL_{40}$) related urinary system. After six days survival of rats, mainly common labeled neurons projecting to the urinary bladder and urinary bladder-related acupoints were identified in spinal cord, medulla, pons and diencephalon by PRV immunohistochemical staining method. First-order PRV labeled neurons projecting to urinary bladder and urinary bladder-related acupoints were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled preganglionic neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in the lateral horn area(sacral parasympathetic nucleus and intermediolateral nucleus), lamina V-X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting to urinary bladder and Wijung($BL_{40}$) was L5-S2 spinal ganglia and T12-L1 spinal ganglia, respectively. In the brainstem, the neurons were labeled most evidently and consistently in the nucleus of tractus solitarius, area postrema, dorsal motor nucleus of vagus nerve, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), C3 adrenalin cells, parapyramidal area(lateral paragigantocellular nucleus), locus coeruleus, subcoeruleus nucleus, A5 cell group, Barrington's nucleus and periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the paraventricular nucleus and a few ones were in the lateral hypothalamic nucleus, posterior hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus, median eminence, perifornical nucleus, periventricular nucleus and suprachiasmatic nucleus. In cerebral cortex, PRV labeled neurons were marked mostly in the frontal cortex, 1,2 area, hind limb area, agranular insular cortex. Immunoreactive neurons to Corticotropin releasiing factor(CRF), Corticotropin releasiing factor-receptor(CRF-R), c-fos and serotonin were a part of labeled areas among the virus-labeled neurons of urinary bladder and Wijung($BL_{40}$). The commonly labeled areas were nucleus tractus solitarius, area postrema, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), locus coeruleus, A5 cell group, Barrington,s nucleus, arcuate nucleus, paraventricular nucleus, frontal cortex 1, 2 area, hind limb, and perirhinal(agranular insular) cortex. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of urinary bladder-relate organs and it was revealed by tracing PRV labeled neurons projecting urinary bladder and urinary bladder-related acupoints. These commonly labeled areas often overlap with the neurons connected with hormones and hormone receptors related to urination.

• Korean Journal of Veterinary Research
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• v.43 no.1
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• pp.11-24
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• 2003

This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.

• Applied Microscopy
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• v.40 no.4
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• pp.211-218
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• 2010

The mammalian ovary is innervated by sympathetic and sensory neurons which contribute to regulating several aspects of ovarian function, including blood flow, steroidogenesis and follicular development. The existence of a neural connection between central neurons and the ovary has been rarely reported, but the mechanism underlying integration of ovarian activity to broader neuroendocrine responses has not been reported. We have now used a viral transneuronal tracing technique combined with a conventional retrograde labeling procedure of CT-HRP to demonstrate that oxytocin-producing neurons of the hypothalamus are synaptically connected to the ovary. Since ovarian activity is suppressed but the activity of oxytocin neurons is increased during breast feeding. Our finding that the oxytocinergic neural connection is likely to provide a direct transsynaptic mechanism by which the central nervous system maintains the state of infertility that accompanies lactation in mammals.

• Korean Journal of Acupuncture
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• v.17 no.1
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• pp.81-100
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• 2000

Transsynaptic tracer이며 신경친화성 virus인 pseudorabies virus(PRV)를 방광(膀胱), 방광유(膀胱兪), 위중(委中) 및 중추(中極)에 주입(注入)한 후 4일간의 생존기간이 경과한 후 희생시켜 면역조직화학침액법(免疫組織化學染色法)에 의하여 뇌척수에 표지된 공통된 영역들을 비교하여 관찰한 결과는 다음과 같다. 1. 방광벽(膀胱壁), 방광유(膀胱兪), 위중(委中) 및 중추(中極)에서 척수에 투사된 영역은 흉수(胸髓), 요수(腰髓) 및 천수(薦髓)에 모두 표지되었으며 공통적으로 표지된 부위는 척수(脊髓)의 층판 IV, V, VII, IX, X영역에 표지되었으나 주로 강하게 표지된 공통된 영역은 층판 VII의 중간외측핵, 가슴기둥 및 층판 X영역이었다. 2. 방광벽(膀胱壁), 방광유(膀胱兪), 위중(委中) 및 중추(中極)에서 뇌(腦)에 투사된 공통된 영역은 연수(延髓)에서는 A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus에서 양성반응을 나타내었다. 솔기핵의 경우 아핵인 불명솔기핵, 창백솔기핵 및 큰솔기핵에서 양성반응을 보였다. 다리뇌에서는 청색반점, Barrington's nucleus, A5세포군 및 삼차신경운동핵에서 양성반응을 보였고, 중뇌에서는 눈돌림신경핵, 눈돌림신경섬유 및 다리핵에서 양성반응을 보였다. 간뇌에서는 시상하부(視床下部)의 뇌실(腦室)곁핵과 시상의 뇌실곁핵에서 양성반응을 보였고 대뇌(大腦)에서는 septal nucleus, 피질(皮質)의 뒷다리영역, 마루엽, 이마엽에서 양성반응을 보였다. 이상의 결과를 종합하면 방광(膀胱)에서 투사되는 뇌척수의 영역과 방광유(膀胱兪)나 위중(委中)에서 투사되는 공통된 표지영역들은 방광(膀胱)과 족태양방광경(足太陽膀胱經) 그리고 그 경락(經絡)의 경혈(經穴)들이 어떤 상관성(相關性)을 가지고 연결(連結)되어 있다는 사실을 실험적으로 알 수 있었다. 특히 방광(膀胱)과 방광유(膀胱兪), 위중(委中)에서 투사된 공통된 표지영역, 즉 배뇨중추인 Barrington's nucleus에 표지되는 것은 내장(內臟)-경락(經絡)이 central autonomic pathway에 의하여 서로 연결되었음을 입증하는 중요한 결과(結果)라고 사려(思慮)된다.