• Title/Summary/Keyword: FGF staining

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Immunohistochemical Study for the Angiogenesis Factors and Vascular Wall Matrix Proteins in Intracranial Aneurysms (뇌동맥류에서 혈관형성 인자와 혈관벽 기질 단백에 대한 면역조직화학적 연구)

  • Kim, Jae Hong;Yim, Man Bin;Lee, Chang Young;Kim, Sang Pyo
    • Journal of Korean Neurosurgical Society
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    • v.29 no.12
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    • pp.1584-1591
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    • 2000
  • Objective : Until now, it has been little known about the biological mechanisms associated with the genesis, growth, and rupture of intracranial aneurysm. This study was performed to investigate and understand a part of these mechanisms. Materials and Methods : Immunohistochemical stains for angiogenesis growth factors(basic fibroblast growth factor (bFGF) and vascular endothelial growth factor(VEGF)) and selected vascular wall matrix proteins(alpha smooth muscle actin(${\alpha}SMA$) and collagen Type IV) were performed in fixed sections from a normal circle of Willis artery which was taken from the autopsy specimen as a control vessel and 17 aneurysmal wall specimens which was taken during surgical clipping of aneurysms. The staining intensity and distribution of immunoreactivity to angiogenesis growth factors and selected wall matrix proteins in control vessel and aneurysmal wall were examined and compared with each other. The difference of staining intensity according to the size of aneurysm was also investigated. Results : There was no immunoreactivity to bFGF and VEGF in the control vessel. bFGF immunoreactivity was exhibited in 15 of 17 aneurysm specimens around smooth muscle cells within the media of aneurysm. VEGF immunoreactivity was also exhibited in all aneurysm specimens in patches or diffusely affecting all layers of the aneurysmal wall. The degrees of intensity of bFGF and VEGF immunoexpression were proportionate roughly to the size of aneurysm. Strong immunoexpression of both factors were noticed in large aneurysm. A regularly arranged and defined band of immunoreactivity of ${\alpha}SMA$ was noticed in the media of the control vessel, whereas diffuse, faint, irregularly arranged ${\alpha}SMA$ was noticed in the aneurysmal wall. A regularly defined band of collagen Type IV immunoreactivity was also noticed in the subendothelium of the control vessel, whereas diffuse disorganized immunoreactivity of collagen Type IV was noticed in the entire wall of the aneurysm. Conclusion : These results indicate substantial evidences of abnormal expression of angiogenesis factors and changes of selected vascular wall matrix proteins in the wall of intracranial aneurysm. The unbalanced changes of angiogenesis factors and vascular wall matrix proteins in the wall of aneurysm may be one of the biological mechanisms for the growth and rupture of aneurysm.

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The Success of Thread-embedding Therapy in Generating Hair Re-growth in Mice Points to Its Possibly Having a Similar Effect in Humans

  • Shin, Hyun Jong;Lee, Dong-Jin;Kwon, Kang;Lee, Ji-Yeon;Ha, Ki-Tae;Lee, Chang-Hyun;Jang, Yong-Suk;Lee, Byung-Wook;Kim, Byung Joo;Jung, Myeong-Ho;Seo, Hyung-Sik;Jeong, Han-Sol
    • Journal of Pharmacopuncture
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    • v.18 no.4
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    • pp.20-25
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    • 2015
  • Objectives: Recently, thread-embedding therapy (TET) has been widely applied in Korean medicine for cosmetic purposes such as reducing skin wrinkles. An inserted thread was reported to have induced continuous stimulation, followed by support for connective tissue regeneration. However, the potential role of TET in hair-growth has not yet been reported. Methods: We designed this study to evaluate whether TET has a hair-growth-promoting effect. C57 black 6 (C57BL/6) mice were divided into three groups: normal saline-treated, minoxidil-treated, and thread-embedded groups. Normal saline or 5% minoxidil was topically sprayed on the dorsal skin of the mice once a day for 16 days. Medical threads were embedded into the dorsal skin of the mice in a single application. Hair growth activity was evaluated by using dermoscopic and microscopic observations. Sections of the dorsal skin were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), fibroblast growth factor-7 (FGF-7), and fibroblast growth factor-5 (FGF-5) were detected by using immunohistochemical staining. A reverse transcription-polymerase chain reaction (RT-PCR) analysis was adopted to measure the messenger RNA (mRNA) expressions of FGF-7 and FGF-5. Results: TET enhanced anagen development in the hair follicles of C57BL/6 mice. The expressions of BrdU and PCNA, both of which imply active cellular proliferation, were increased by using TET. Moreover, TET increased the expression of FGF-7, an anagen-inducing growth factor, while decreasing the expression of FGF-5, an anagen-cessation growth factor, both at the protein and the mRNA levels. Conclusion: TET enhanced hair re-growth in C57BL/6 mice. TET regulated the expressions of anagen-associated growth factors and activated the proliferation of hair follicular cells in depilated skin lesions. Considering its long-lasting effect, TET may be a good alternative therapeutic for the treatment of alopecia.

Immunohistochemical Analysis of the Bullae in Patients with Primary Spontaneous Pneumothorax (원발성 기흉환자 폐기포의 면역조직화학적 분석)

  • 김광호;윤용한;김정택;백완기;김현태;김영삼;손국희;한혜승;박광원
    • Journal of Chest Surgery
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    • v.36 no.2
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    • pp.86-90
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    • 2003
  • Bulla is an air-filled space within the lung parenchyma resulting from deterioration of the alveolar tissue. Molecular mechanism of the formation of the bulla is not well described. Fibroblast growth factor(FGF)-7, bone morphogenetic protein(BMP) receptor, and transforming growth factor(TGF)-$\beta$ receptor are known to have a stimulatory or inhibitory role in the lung formation. We investigated to see if these growth factor or cytokine receptors are involved in the bulla formation by immunohistochemical staining of bullous lung tissues from patients with primary spontaneous pneumothorax. Material and Method: Bullous lung tissues were obtained from 31 patients with primary spontaneous pneumothorax, including 30 males and 1 female from 15 to 39 years old. The bullous tissues were obtained by video-thoracoscopic surgery and/or mini-thoracotomy and fixed in formalin. Blocks of the specimens were embedded with paraffin and cut into 5-6 ${\mu}{\textrm}{m}$ thick slices. The sections were deparaffinized and hydrated and then incubated with primary antibodies against FGF-7, BMP-RII, or TGF-RII. Result: Of the 31 patients, 24 were TGF-RII positive including 18 strong and 6 weak positives. Observation with high magnification showed that strong immunostaining was detected in the boundary region between bullous and normal lung tissues. In contrast, all of the sections were negative with FGF-7 or BMP-RII antibodies. Conclusion: These results suggest that overexpression of TGF- P RII may be involved in the formation of bulla, although further molecular studies are needed to find out more detailed molecular mechanisms.

Effect of Herbal Decoction for Sitz Bath on Dermoepidermal Recovery to Wound Tissue in Rats (한방좌욕제의 창상치유 효과에 대한 실험적 연구)

  • Kwon, Su-Kyung;Choi, Min-Sun;Yoon, Sang-Ho;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.1
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    • pp.30-41
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    • 2010
  • Purpose: This study was to observe the effect of "herbal decoction for sitz bath" on dermoepidermal recovery to wound tissue in rat's skin. Methods: The samples were assigned to 3 groups: control group : without any treatment, positive control group : potarose 10% solution, experiment group : herbal decoction for sitz bath. We made the open wound of $2{\times}2cm^2$ size that cut deep into the dermis. Treating the open wound for 17 days, we observed the size of the wound diminishing. On 17th days, the cell viability was measured by MTT assay. The effect anti-inflammatory and dermoepidermal recovery were examined by H&E staining, immunohystochemical staining for MIP-2, FGF. Results: The experiment group showed more recovery from the open wound comparing the control group and the positive control group on 10th days after wounding. But there was not remarkable difference between the experiment and positive control group after 17th days post-wounding. The number of MIP-2 positive reacted cell were significantly decreased and that of FGF positive reacted cell were significantly increased than positive control group at 17th days. Conclusion: According to these results, we finally concluded that "herbal decoction for sitz bath" could be effective in recovery to wound tissue.

The Effect of Ulmus Root-bark Dressing in Fibroblast Growth Factor and Vascular Endothelial Growth Factor of Induced Pressure Ulcer in Rats (느릅나무 근피드레싱이 쥐에 유발된 욕창의 섬유아세포성장인자와 혈관내피성장인자에 미치는 효과)

  • Na, Yeon Kyung
    • Journal of Korean Biological Nursing Science
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    • v.15 no.4
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    • pp.257-263
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    • 2013
  • Purpose: The purpose of this study was to investigate the effect of Ulmus root-bark dressing in fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) of induced pressure ulcers in rats. Methods: 54 male Sprague-Dawley rats were used and randomly divided into 2 groups. The rats were anesthetized and pressure ulcers were induced at 140 mmHg for three hours, using a personally-designed pressing apparatus. Ulmus dressing was applied in the experimental group (n=27) and saline gauze dressing in the control group (n=27). Each of the dressings was changed every other day, and after a month, the wounds were examined by microscopy biweekly for 20 weeks. Results: After 4 weeks, the epidermis of the wounds was recovered, but inflammatory infiltration of the dermis was remained. After 6 weeks, inflammatory cells were diminished and the number of capillaries was decreased. Examined by immunofluorescence staining, the FGF positive cells in the experimental group changed negatively after 18 weeks, but the control group still existed even after 20 weeks. VEGF positive cells in the experimental group also changed negatively after 20 weeks, but the control group still existed. Conclusion: The findings of this study demonstrate that Ulmus dressing is effective in minimizing scar formation and inflammatory reaction by decreasing FGF and VEGF in the terminal phase of wound healing.

Hair-growth Promoting Effect of Microneedle Roller Therapy (미세침요법의 모발성장효과)

  • Lee, Chang Hyun;Lee, Ji Yeon;Shin, Hyun Jong;Ha, Ki Tae;Seo, Hyung Sik;Jeong, Han Sol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.1
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    • pp.16-21
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    • 2014
  • Micro needle roller therapy has been used for cosmetic purposes, such as reducing skin winkles and improving elasticity of skin. It is claimed that micro needle roller therapy has potentials for connective tissue regeneration by facilitating collagen synthesis. Therefore, there seems to be a possibility that connective tissue regenerating potential of micro needle roller therapy could influence the hair growth cycle. This study, we investigated the hair growth-promoting effects of micro needle roller therapy. C57BL/6 mice were devided into three groups as follows: normal saline-treated, minoxidil-treated, and micro needle roller therapy-received group. Hair growth activity was evaluated by handscopic and microscopic observations. Sections of dorsal skin were stained with hematoxylin and eosin. Expression of BrdU, FGF, and VEGF was detected by immunohistochemical staining. Micro needle roller therapy enhanced the development of hair follicle during anagen. Immunohistochemical analysis revealed that micro neeld roller therapy incresed the expression of BrdU and FGF in the hair follicles of C57BL/6 mice. Furthermore, micro needle roller therapy upregulated mRNA expression of VEGFR-2, FGF-2, EGF - growth factors that play a central role in hair follicle development during anagen. These results suggest that Micro needle roller therapy can potentially be used for the treatment of alopecia.

Cornu cervi pantotrichum Pharmacopuncture Solution Facilitate Hair Growth in C57BL/6 Mice

  • Lee, Seon-Yong;Lee, Dong-Jin;Kwon, Kang;Lee, Chang-Hyun;Shin, Hyun Jong;Kim, Jai Eun;Ha, Ki-Tae;Jeong, Han-Sol;Seo, Hyung-Sik
    • Journal of Pharmacopuncture
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    • v.19 no.2
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    • pp.122-128
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    • 2016
  • Objectives: Cornu cervi pantotrichum (CCP) has been widely used in Korean and China, as an anti-fatigue, anti-aging, and tonic agent to enhance the functions of the reproductive and the immune systems. Because CCP has various growth factors that play important roles in the development of hair follicles, we examined whether CCP pharmacopuncture solution (CCPPS) was capable of promoting hair growth in an animal model. Methods: One day after hair depilation, CCPPS were topically applied to the dorsal skin of C57BL/6 mice once a day for 15 days. Hair growth activity was evaluated by using macro- and microscopic observations. Dorsal skin tissues were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), and fibroblast growth factor (FGF)-7 were examined by using immunohistochemical staining. A reverse transcription polymerase chain reaction (RT-PCR) analysis was also conducted to measure the messenger RNA (mRNA) expression of FGF-7. Results: CCPPS induced more active hair growth than normal saline. Histologic analysis showed enlargement of the dermal papilla, elongation of the hair shaft, and expansion of hair thickness in CCPPS treated mice, indicating that CCPPS effectively induced the development of anagen. CCPPS treatment markedly increased the expressions of BrdU and PCNA in the hair follicles of C57BL/6 mice. In addition, CCPPS up regulated the expression of FGF-7, which plays an important role in the development of hair follicles. Conclusion: These results reveal that CCPPS facilitates hair re-growth by proliferation of hair follicular cells and up-regulation of FGF-7 and suggest that CCPPS can potentially be applied as an alternative treatment for patients with alopecia.

Human Embryonic Stem Cell Transplantation in Parkinson′s Disease (PD) Animal Model: II. In Vivo Transplantation in Normal or PD Rat Brain

  • Choe Gyeong-Hui;Ju Wan-Seok;Kim Yong-Sik;Kim Eun-Yeong;Park Se-Pil;Im Jin-Ho
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.19-19
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    • 2002
  • This study was to examine whether the in vitro differentiated neural cells derived from human embryonic stem (hES, MB03) cells can be survived and expressed tyrosin hydroxylase(TH) in grafted normal or PD rat brain. To differentiate in vitro into neural cells, embryoid bodies (EB: for 5 days, without mitogen) were formed from hES cells, neural progenitor cells(neurosphere, for 7-10 days, 20 ng/㎖ of bFGF added N2 medium) were produced from EB, and then finally neurospheres were differentiated into mature neuron cells in N2 medium(without bFGF) for 2 weeks. In normal rat brain, neural progenitor cells or mature neuron cells (1×10/sup 7/ cells/㎖) were grafted to the striatum of normal rats. After 2 weeks, when the survival of grafted hES cells was examined by immunohistochemical analysis, the neural progenitor cell group indicated higher BrdU, NeuN+, MAP2+ and GFAP+ than mature neuron cell group in grafted sites of normal rats. This result demonstrated that the in vivo differentiation of grafted hES cells be increased simultaneously in both of neuronal and glial cell type. Also, neural progenitor cell grafted normal rats expressed more TH pattern than mature neuron cells. Based on this data, as a preliminary test, when the neural progenitor cells were grafted into the striatum of 6-hydroxydopamine lesioned PD rats, we confirmed the cell survival (by double staining of Nissl and NeuN) and TH expression. This result suggested that in vitro differentiated neural progenitor cells derived from hES cells are more usable than mature neuron cells for the neural cell grafting in animal model and those grafted cells were survived and expressed TH in normal or PD rat brain.

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Effect of Parthenogenetic Mouse Embryonic Stem Cell (PmES) in the Mouse Model of Huntington′s Disease

  • 이창현;김용식;이영재;김은영;길광수;정길생;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.80-80
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    • 2003
  • Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor, cognitive, and psychiatric symptoms, accompanied by marked cell death in the striatum and cortex. Stereotaxic injection of quinolinic acid (QA) into striatum results in a degeneration of GABAergic neurons and exhibits abnormal motor behaviors typical of the illness. The objective of this study was carried out to obtain basic information about whether parthenogenetic mouse embryonic stem (PmES) cells are suitable for cell replacement therapy of HD. To establish PmES cell lines, hybrid F1 (C57BL/6xCBA/N) mouse oocytes were treated with 7% ethanol for 5 min and cytochalasin-B for 4 hr to initiate spontaneous cleavage. Thus established PmES cells were induced to differentiate using bFGF (20ng/ml) followed by selection of neuronal precursor cells for 8 days in N2 medium. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days, then a final differentiation step in N2 medium for 7 days. To establish recipient animal models of HD, young adult mice (7 weeks age ICR mice) were lesioned unilaterally with a stereotaxic injection of QA (60 nM) into the striatum and the rotational behavior of the animals was tested using apomorphine (0.1mg/kg, IP) 7 days after the induction of lesion. Animals rotating more than 120 turns per hour were selected and the differentiated PmES cells (1$\times$10$^4$cells/ul) were implanted into striatum. Four weeks after the graft, immunohistochemical studies revealed the presence of cells reactive to anti-NeuN antibody. However, only a slight improvement of motor behavior was observed. By Nissl staining, cell mass resembling tumor was found at the graft site and near cortex which may explain the slight behavioral improvement. Detailed experiment on cell viability, differentiation and migration explanted in vivo is currently being studied.

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Hepatocyte Growth Factor is the Key Cytokine in Stimulating Potential Stem Cells in the Cord Blood into Hepatic Lineage Cells

  • Ryu, Kyung-Ha;Cho, Su-Jin;Woo, So-Youn;Seoh, Ju-Young;Jung, Yun-Jae;Han, Ho-Seong
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.117-123
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    • 2007
  • Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.