• Title/Summary/Keyword: Immunostain

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The Effect of Woohwangcheongsim-won on Delayed Neuronal Death in Hypoxia (저산소증으로 유발된 지연성 신경세포사에 우황청심원이 미치는 영향)

  • 김민석;정승현;신길조;문일수;이원철
    • The Journal of Korean Medicine
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    • v.23 no.3
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    • pp.145-163
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    • 2002
  • Objectives: The purpose of this investigation was to evaluate the effects of Woohwangcheongsim-won and to study the mechanism for neuronal death protection in hypoxia with Embryonic day 20 (E20) cortical cells of a rat (Sprague Dawley). Methods: E20 cortical cells were dissociated in neurobasal media and grown for 14 days in vitro (DIV). On 14 DIV, Woohwangcheongsim-won was added to the culture media for 24 hrs or 72 hrs. On 17 DIV, cells were given a hypoxic shock and further incubated in normoxia for another three days. On 20 DIV, Woohwangcheongsim-won's effects for neuronal death protection were evaluated by LDH assay, propidium iodide stain and phospho-H2AX immunostain and the mechanisms were studied by Bcl-2, Bak, Bax, caspase family, PKCα, ca1pain I. Results & Conclusions : 1. This study indicated that Woohwangcheongsim-won's effects for neuronal death protection in hypoxia were confirmed by LDH assay, propidium iodide stain and phospho-H2AX immunostain in culture method of Embryonic day 20(E20) cortical neuroblasts. 2. Woohwangcheongsim-won's mechanisms for neuronal death protection in hypoxia are to reduce the membrane damage fraction, to restrain DNA truncate, to restrain inflow of cytochrome c into cellularity caused by Bak diminution, to reduce the caspase cascade intiator caspase-8 and the effector caspase-3, to reduce the calpain I activity and to increase PKCand its activity in the membrane fraction. (J Korean Oriental Moo 2002;23(3):145~163)

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Effect of EGF on In Vitro Oocyte Maturation and Embryo Development and Expression of EGF mRNA in Bovine Oocytes and Embryo II. Detection of Epidermal Growth Factor mRNA in bovine Ova during In Vitro Maturation and after Fertilization In Vitro

  • Kim, Kwang-Sig;Kim, Chang-Keun;Chung, Yung-Chai;Hwang, Seong-Soo;Chang, Won-Kyong;Cheong, Il-Cheong;Park, Jin-Ki;Min, Kwan-Sik;Lee, Yun-Keun
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.29-29
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    • 2001
  • This study was carried out to examine, by the reverse transcription chain reaction(RT-PCR)and Immunostain assays, epidermal growth factor mRNA expression in bovine ova during oocyte maturation in vitro(0-2lh)and after fertilization in vitro(6-144hr: zygotes to blastocysts). In this study, the transcripts of EGF was detected in oocytes using primers for EGF. Transcripts for EGF mRNA was not detected in oocytes through in vitro maturation. But EGF mRNA were present after fertilization up to the 2-cell stage and the blastocyst stage. The highest mRNA levels in 4-cell stage embryos were decreased at 8cell stage and then reincreased upto morulae and blastocysts. The results of this study showed EGF mRNA are present in embryo after fertilization and this factors are involved in the regulation of bovine embryo development.

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Effects of TGF ${\beta}_1$ on the Growth and Biochemical Changes in Cultured Rat Glial Cells (Transforming growth factor ${\beta}_1$이 배양랫트 신경교세포의 성장 및 생화학적 변화에 미치는 영향)

  • Kim, Yong-Sik;Youn, Yong-Ha;Park, Nan-Hyang;Park, Chan-Woong
    • The Korean Journal of Pharmacology
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    • v.30 no.2
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    • pp.167-179
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    • 1994
  • Recent evidence indicates that glial cells have a wide range of funtions which are critical for maintaining a balanced homeostatic environment in the central nervous system(CNS) peripheral nervous system(PNS). Morever, astrocytes are known to participate in the tissue repair and neuroimmunologic events within the CNS through many kinds of growth factors and cytokines. We investigated the effect of $TGF\;{\beta}_1$, on the growth and biochemical changes of rat glial cells in culture. The proliferative effect was determined by $^3H-thymidine$ uptake and the double immunostain with anti-cell-specific marker and anti-Bromodeoxyuridine(BrdU) antibody. To check the effect of biochemical changes we compared the amounts of glial fibrillar acidic protein(GFAP) and the activity of glutamine synthetase(GS) in astrocyte. And the amounts of myelin basic protein and the activity of 2',3'-cyclic nucleotide phosphohydrolase(CNPase) were measured in oligodendrocyte and the amounts of peripheral myelin in Schwann cell. When $TGF\;{\beta}_1$, was treated for 2 days with cultured glial cell, $TGF\;{\beta}_1$, decreased the $^3H-thymidine$ uptake and proliferation index of double immunostain of astrocytes, which indicates the inhibition of astroglial DNA synthesis, but stimulated the growth of Schwann cell. Also, $TGF\;{\beta}_1$, decrease the GS activity and increased the amounts of GFAP in astrocyte. In the case of Schwann cells the amounts of peripheral myelin was increased when treated with $TGF\;{\beta}_1$. However, $TGF\;{\beta}_1$, didn't show any effect on the proliferation and biochemical changes in oligodendrocyte. These results suggest that $TGF\;{\beta}_1$, might have a critical action in the regulation of proliferation and biochemical changes in glial cells, especially astrocyte.

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Chondrogenesis of Mesenchymal Stem Cell Derived form Canine Adipose Tissue

  • Lee, Byung-Joo;Wang, Soo-Geun;Seo, Cheol-Ju;Lee, Jin-Chun;Jung, Jin-Sup;Lee, Ryang-Hwa
    • Proceedings of the KSLP Conference
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    • 2003.11a
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    • pp.183-183
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    • 2003
  • Background and Objectives : Cartilage reconstruction is one of medical issue in otolaryngology. Tissue engineering is presently being utilized in part of cartilage repair. Sources of cells for tissue engineering are chondrocyte from mature cartilage and bone marrow mesenchymal stem cells that are able to differentiate into chondrocyte. Recent studies have shown that adipose tissue have mesenchymal stem cells which can differentiate into adipogenic, chondrogenic myogenic osteogenic cells and neural cell in vitro. In this study, we have examined chondrogenic potential of the canine adipose tissue-derived mesenchymal stem cell(ATSC). Materials and Methods : We harvested canine adipose tissue from inguinal area. ATSCs were enzymatically released from canine adipose tissue. Under appropriate culture conditions, ATSCs were induced to differentiate into the chondrocyte lineages using micromass culture technique. We used immunostain to type II collagen and toluidine blue stain to confirm chondrogenic differentiation of ATSCs. Results : We could isolate ATSCs from canine adipose tissue. ATSCs expressed CD29 and CD44 which are specific surface markers of mesenchymal stem cell. ATSCs differentiated into micromass that has positive response to immunostain of type II collagen and toluidine blue stain. Conclusion : In vitro, ATSCs differentiated into cells that have characteristic cartilage matrix molecules in the presence of lineage-specific induction factors. Adipose tissue may represent an alternative source to bone marrow-derived MSCs.

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An OTHBVS Cell Line Expresses the Human HBV Middle S Protein

  • Park, Sung-Gyoo;Guhung Jung
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.86-89
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    • 1999
  • An OTHBVS cell line from HepG2 was established. This cell line stably expresses the human hepatitis B virus (HBV) middle S protein that includes the preS2 region which is important for HBV particle entry into the hepatocyte. To establish this cell line, the middle S open reading frame (ORF), with a promoter located in the 5' region and enhancer located in the 3' region, was cloned downstream from the metallothionine (MT) promoter of the OT1529 vector. In this vector, expression of the middle S protein was constructed to be regulated by its own promoter and enhancer. Expression of the large S protein which contains the preS1 region in addition to the middle S protein was designed to be regulated by the MT promoter. When extracts of OTHBVS cells were examined with an S protein detection kit (RPHA, Korea Green Cross Co.), an S protein was detected. Total mRNA of OTHBVS cell examined by northern blot analysis with an S ORF probe revealed small/middle S transcripts (2.1 kb). When the MT promoter was induced by Zn, large S transcripts (2.4 kb) were detected. The GP36 and GP33 middle S proteins were presumably detected, but large S proteins were not detected by immunostain analysis using anti-preS2 antibody.

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Localization of Amygdaloid Nucleus Innervating the Stomach Using Genetically Engineered PRV-BaBlu in Rat Brain (유전자 조작된 PRV-BaBlu를 이용한 흰쥐 위 신경지배 편도핵의 동정)

  • Song, Ju-Min
    • The Journal of Korean Physical Therapy
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    • v.23 no.5
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    • pp.35-41
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    • 2011
  • Purpose: This study was carried out to investigate the spatiotemporal localization of the amygdaloid nucleus innervating the rat stomach using PRV-BaBlu, which has been known to be an excellent type of neurotracer with the ability to transpass the neuronalsynaptic cleft. Methods: Ninety Sprague-Dawley rats (250~300 g) that were injected with PRV-BaBlu into the stomach were randomly divided into 3, 4 and 5 day groups (each group n=30). $2{\mu}l$ of PRV-BaBlu, a genetically modified strain of PRV-Bartha with the lac-Z gene,was injected into the rat stomach and immunostained with a mouse anti-${\beta}$-galactosidase at 3, 4 and 5 days after the virus injection. Results: The PRV-BaBlu infected the neurons in the amygdaloid nucleus, and the degree of viral infection in experimental animals showed a tendency to increase significantly with time (p<0.05). The neurons between the left and right amygdaloid nucleus significantly differ (p<0.05). Conclusion: This showed that PRV-BaBlu was an excellent neurotracer for localizing the amygdaloid nucleus, and the amygdaloid nucleus has a sensory input and motor output on stomach movement, influencing emotional behavior.

Effects of Saengjinyanghyutang-gamibang and Rosa rugosa on the Pancreas in the Streptozotocin Treated Mice for Induction of Diabetes (생진양혈탕가미방과 매괴화가 Streptozotocin으로 유발된 고혈당 생쥐의 췌장에 미치는 영향)

  • 홍광표;나창수;장경선;김희철;박민희;김정상
    • The Journal of Korean Medicine
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    • v.22 no.4
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    • pp.79-89
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    • 2001
  • Objectives : This study has been carried out to investigate the effects of the Saengjinyanghyutang-gamibang and Rosa rugosa extracts on pancreas of the hyperglycemia mice induced with streptozotocin. Methods : We examined immunohistochemistry for insulin and COX-2, ultrastructural changes of acini by electron microscope, and changes of the blood glucose and BUN levels. Results : The ${\beta}-cells$ on Langerhnan's islet were destructed by administration of streptozotocin, so that few insulin-positive cells were observed in the control group. However, a lot of insulin-positive cells were observed in the experimental groups. These cells had recovered from the damage. As a result of COX-2 immunostain, COX-2 expression were highest in the control group other than the Saengjinyanghyutang-gamibang and Rosa rugosa extracts administered groups. As the electron microscopical observation, the centroacinar cells and acini of pancreas were destructed or damaged by administration of streptozotocin in the control group, but these recovered !Tom the damage in the other experimental groups. The levels of serum glucose were decreased remarkably on the Rosa rugosa and Saengjinyanghyutang-gamibang extracts administered groups compared with control group. Conclusions : These results suggest that administration of the Rosa rugosa and Saengjinyanghyultang-gamibang extracts to the mice reduced the damage induced by streptozotocin.

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A Case of Malignant Triton Tumor on Supraclavicular Area (쇄골 상부에 발생한 악성 Triton 종양 1예)

  • Lim, Sang Ho;Park, Hee Tack;Hong, Ki Hwan
    • Korean Journal of Head & Neck Oncology
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    • v.29 no.2
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    • pp.54-57
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    • 2013
  • Malignant triton tumor(MTT) is a rare type of malignant peripheral nerve sheath tumor(MPNST) with focal rhabdomyoblastic differentiation. MTT constitutes about 5% of all MPNSTs and described the first case of a MTT in a patient with Von Recklinghausen disease by Masson in 1932. MTT is commonly seen in the head, neck, extremities and trunk. It can occur in sporadic form or over a setting of neurofibromatosis-1(NF-1). The diagnosis can be confirmed based on morphologic grounds supported by an immunostain such as S-100 protein. Desmin, myo-D1 and myogenin are immunostains positive for rhabdomyoblasts. MTT has an aggressive biological behavior so prognosis of this rare and highly malignant tumor is poor and optimal treatment remains unclear. But modern treatment consisted of radical excision and postoperative radiotherapy has improved the prognosis of such cases.

SDS-PAGE and Immunoblot Patterns of Echinostoma hortense in Experimentally Infected Rats (SDS-PAGE 및 면역이적법에 의한 호르텐스극구흡충 항원분획과 항체반응 양상)

  • Yong-Suk Ryang;Yoon-Kyung Jo;Ji-Sook Lee
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.73-76
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    • 1998
  • The authors characterized the antigen proteins and some specific antibodies from Echinostoma hortense. Crude antigen extracted from E. hortense worm was analyzed by SDS-PAGE of the crude antigen showed 46 profiles between 200.2 - 8.2kDa, among which 200.2, 107.9, 86.8, 75, 69.8, 46.8, 43.5, 34.5, 20.9, 13.6, 12.6, 11.7, and 8.2kDa, protein profiles were strong. EITB resolved the specific IgG antibody into 17 profiles between 193 - 13.7kDa, among which 198, 123.4, 100.8, 91.1, 88.1, 62.8, 34.2, 32, 29.9, 18, 15.7, 13.7kDa profiles showed strong immunostain.

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Neurotropism and Expression Pattern of lacZ Inserted PRV-Bartha in Geniculohypothalamic Tract Tracing (lacZ가 삽입된 PRV-Bartha 종의 신경친화성 및 무릎시상하부로 추적시 발현양상 분석)

  • Kim, Jin-Sang;Park, Eun-Se;Cheon, Song-Hee;Kim, Min-Hee;Bang, Hyun-Soo;Kwon, Young-Shil;Lee, Bong-Hee;Kim, Young-Chul
    • Toxicological Research
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    • v.22 no.4
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    • pp.403-409
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    • 2006
  • To localize the connection between intergeniculate nucleus and suprachisasmatic nucleus through geniculohypothalamic tract in postnatal mongolian gerbil, we injected lacZ inserted PRV-Bartha strain into suprachiasrnatic nucleus and tried to immunostain against it with Rb134 and mouse $anti-{\beta}-galactosidase$. The numbers of immunoreactive neurons in intergeniculate leaflet were $8{\pm}3.2$ in P1 Period, $10{\pm}4.1$ in P3 Period and $13{\pm}6.2$ in P7 Period, and was statistically significant (p<0.05) and had tendency to increase with time consuming. The results showed that intergeniculate leaflet had projected some axons into suprachiasrnatic nucleus through geniculohyptothalamic tract in postnatal mongolian gerbil. But we could not exclude the possibility of direct projections from dorsal and ventral geniculate nuclei into suprachisamatic nucleus completely.