• Title/Summary/Keyword: Re-RT

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Expression and Localization of Heat Shock Protein 70 in Frozen-Thawed IVF and Nuclear Transfrred Bovine Embryos

  • Park, Y.J;S.J Song;J.T Do;B.S Yoon;Kim, A.J;K.S Chung;Lee, H.T
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.78-78
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    • 2002
  • The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

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Expression of Serotonin(5-HT) Receptor Isotypes in Reproductive Organs of Male Rat (수컷 흰쥐 생식기관에서의 세로토닌 수용체 아형 유전자 발현)

  • 이성호
    • Development and Reproduction
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    • v.6 no.2
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    • pp.111-115
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    • 2002
  • 5-Hydroxytryptamine(5-HT; serotonin) system has been implicated in the modulation of male sexual behaviors and the secretion of reproductive hormones. In human males, selective serotonin re-uptake inhibitors(SSRIs) are known to improve the major male sexual dysfunction, premature ejaculation, through the central nervous system-mediated pathways. As numerous hormone and local factors, 5-HT may have peripheral role in the regulation of male sexual function. The expression of 5-HT receptor subtypes in the target tissue, however, has not been explored yet. The present study was undertaken to test whether the 5-HT receptor subtypes are expressed in the reproductive tissues of male rat, especially in ejaculatory machinery such as seminal vesicle and vas deferens. To do this, reverse transcription-polymerase chain reaction(RT-PCR) and Southern blot analysis were employed. The transcripts for the 1A, 1B and 2C subtypes of 5-HT receptor were amplified in all the tested tissues. The present study demonstrated the expression of 5-HT receptor in the rat ejaculatory machinery, suggesting that 5-HT may play a pivotal role in the male sexual function via not only central pathway but also peripheral route. Further study on the receptor subtype-specific effect and their harmonized mode of action will be needed to establish the understanding of ejaculation mechanism and drug design.

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The Impact of Bladder Volume on Acute Urinary Toxicity during Radiation Therapy for Prostate Cancer (전립선암의 방사선치료시 방광 부피가 비뇨기계 부작용에 미치는 영향)

  • Lee, Ji-Hae;Suh, Hyun-Suk;Lee, Kyung-Ja;Lee, Re-Na;Kim, Myung-Soo
    • Radiation Oncology Journal
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    • v.26 no.4
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    • pp.237-246
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    • 2008
  • Purpose: Three-dimensional conformal radiation therapy (3DCRT) and intensity-modulated radiation therapy (IMRT) were found to reduce the incidence of acute and late rectal toxicity compared with conventional radiation therapy (RT), although acute and late urinary toxicities were not reduced significantly. Acute urinary toxicity, even at a low-grade, not only has an impact on a patient's quality of life, but also can be used as a predictor for chronic urinary toxicity. With bladder filling, part of the bladder moves away from the radiation field, resulting in a small irradiated bladder volume; hence, urinary toxicity can be decreased. The purpose of this study is to evaluate the impact of bladder volume on acute urinary toxicity during RT in patients with prostate cancer. Materials and Methods: Forty two patients diagnosed with prostate cancer were treated by 3DCRT and of these, 21 patients made up a control group treated without any instruction to control the bladder volume. The remaining 21 patients in the experimental group were treated with a full bladder after drinking 450 mL of water an hour before treatment. We measured the bladder volume by CT and ultrasound at simulation to validate the accuracy of ultrasound. During the treatment period, we measured bladder volume weekly by ultrasound, for the experimental group, to evaluate the variation of the bladder volume. Results: A significant correlation between the bladder volume measured by CT and ultrasound was observed. The bladder volume in the experimental group varied with each patient despite drinking the same amount of water. Although weekly variations of the bladder volume were very high, larger initial CT volumes were associated with larger mean weekly bladder volumes. The mean bladder volume was $299{\pm}155\;mL$ in the experimental group, as opposed to $187{\pm}155\;mL$ in the control group. Patients in experimental group experienced less acute urinary toxicities than in control group, but the difference was not statistically significant. A trend of reduced toxicity was observed with the increase of CT bladder volume. In patients with bladder volumes greater than 150 mL at simulation, toxicity rates of all grades were significantly lower than in patients with bladder volume less than 150 mL. Also, patients with a mean bladder volume larger than 100 mL during treatment showed a slightly reduced Grade 1 urinary toxicity rate compared to patients with a mean bladder volume smaller than 100 mL. Conclusion: Despite the large variability in bladder volume during the treatment period, treating patients with a full bladder reduced acute urinary toxicities in patients with prostate cancer. We recommend that patients with prostate cancer undergo treatment with a full bladder.

Selective Expansion of TCR $V{\beta}3$+CD4+T Cells in Collagen-induced Arthritis in DBA/1 Mice (콜라겐 유도 관절염에서 콜라겐 항원 특이 $V{\beta}3$+CD4+T 세포의 선택적 증식)

  • Lee, Jae-Seon;Cho, Mi-La;Lee, Jung-Eun;Min, So-Youn;Yoon, Chong-Hyeon;Kim, Wan-Uk;Min, Jun-Ki;Park, Sung-Hwan;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.78-88
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    • 2005
  • Background: Collagen-induced arthritis (CIA) in mice is animal model of autoimmune disease known as rheumatic arthritis in human. We investigated CII-specific CD4+ T cell receptor usage in CIA mice. Methods: In CIA model, draining lymph node (dLN) CD4+ T cells and splenocytes at $3^{rd},\;5^{th},\;8^{th}$ week, we investigated CII-specific T cell proliferation, production of IL-17, IFN-${\gamma}$, TNF-${\alpha}$, IL-4 and IL-10. And we also performed anti-CII IgG Ab measurements in serum level, TCRV ${\beta}$ usage and T cell clonality with RT-PCR-SSCP analysis. Also, we performed proliferative response against CII when CII-specific T cell subset is deleted. Results: CIA mice showed more increase in the serum level of anti-CII IgG than normal mice after induction of arthritis. And the level of anti-CII IgG2a in CIA mice was increased after $3^{rd}$ week after primary immunization, while anti-CII IgG1 was decreased. Draining LN CD4+ T cells have proliferated against CII stimulation at $3^{rd}$ week after $1^{st}$immunization. CD4+T cells derived from dLN of CIA mice produced proinflammatory cytokine IFN-${\gamma}$, IL-17 etc. Draining LN CD4 T cells of CIA presented higher proportion of CD4+V ${\beta}3$+subset compared to those of normal mice at $3^{rd}$ week after $1^{st}$ immunization, and they were increased in proportion by CII stimulation. Draining LN CD4+ T cells without TCRV ${\beta}3+/V{\beta}8.1/8.2+/V{\beta}$10b+cells were not responsive against CII stimulation. But, CII-reactive response of TCRV ${\beta}3-/V{\beta}8.1/8.2-/V{\beta}$10b- T cells was recovered when $V{\beta}3+$ T cells were added in culture. Conclusion: Our results indicate that CD4+$V{\beta}3+$ T cells are selectively expanded in dLN of CIA mice, and their recovery upon CII re-stimulation in vitro, as well as the production Th1-type cytokines, may play pivotal role in CIA pathogenesis.

The Effects of Sibjeondaebotanggamibang on the Treating of Wound (십전대보탕가미방(十全大補湯加味方)의 창상(創傷) 치유(治癒) 효과(效果))

  • Jung, Hoon;Lee, Hyun-Jae;Kim, Binnara;Lee, Chi-Ho;Lee, Eun-Jung;Heo, Dong-Seok;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.3
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    • pp.51-69
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    • 2014
  • Objectives This study was aimed to investigate the effects of Sibjeondaebotanggamibang (SJT) on the wound-induced rats. Methods It was observed the effects of anti-oxidation and anti-inflammation by using of lipopolysaccharide (LPS)-treated RAW 264.7 cells. For the observing on SJT anti-oxidation, it needed to mesure the total amount of polyphenol, DPPH scavenging ability, ABTS scavenging ability and the value of ROS production. In order to observe on the anti-inflammation of SJT, it was mesured the value of No and Cytokine (TNF-${\alpha}$, IL-$1{\beta}$, IL-6). It needed to make a scar (around $2{\times}2cm^2$) on the top of the fascia in the back of the rats and then the rats were divided into 4 groups (n=6). Control group was not treated at all, whereas SJ group was orally medicated SJT, Terra group was per-cutaneously applied Terramycin, and SJ+Terra group was both orally medicated SJT and percutaneously applied Terramycin per day for three weeks. The size of wound was measured with Digimatic Caliper and the blood samples (WBC, neutrophil, monocyte, lymphocyte) were analyzed using Minos-ST, which were collected by cardiac puncture. The effect on inflammatory cytokine (TNF-${\alpha}$, IL-$1{\beta}$, IL-6), immunological cells in synovial fluid was measured. To measure the wound factor expressed by wounded skin sample, we extracted RNA and to investigate MMP-1,2,9 we used RT-PCR. For performing histopathological examinations, we paralyzed the rats by ether, and extracted wounded skin tissues, which were measured by H & E, and monitored on the optical microscope. Results 1. DPPH and ABTS scavenging activity of SJT was increased concentration-dependantly, and ROS scavenging activity was significantly increased (10, $100{\mu}g/ml$). 2. NO production was significantly reduced in SJT treated cells ($100{\mu}g/ml$), both TNF-${\alpha}$ and IL-6 in SJT treated cells (1, 10, $100{\mu}g/ml$), and IL-$1{\beta}$ in SJT treated cells (1, $100{\mu}g/ml$). 1. The size of wound was significantly decreasing in SJ group, Terra group, SJ+Terra group. 2. WBC was significantly reduced in SJ and SJ+Terra group, monocyte in SJ+Terra group. Neutrophil was also reduced in SJ, SJ+Terra group but meaningless. 3. TNF-${\alpha}$ and IL-6 were significantly reduced in SJ group, Terra group, SJ+Terra group, and IL-$1{\beta}$ in SJ+Terra group. 4. mRNA expression in MMP-1 was significantly reduced in SJ group. 5. Collegan production and chronic inflammation were significantly decreased in SJ group, Terra group, SJ+Terra groups. Re-epithelization on the skin in Terra group, SJ+Terra groups was decreased. Conclusions According to this in vitro experiment, Sibjeondaebotanggamibang (SJT) has the effects of anti-oxidative and anti-inflammatory. By in vivo experiment, SJT has the effects of anti-inflammatory. Moreover, the progress of recovery was found visually, heamatologically, genetically and histopathologically. In conclusion, it could be thought that SJT has effect on the treating of wound.

Amended Soil with Biopolymer Positively Affects the Growth of Camelina sativa L. Under Drought Stress (가뭄 조건 하에서 바이오폴리머 혼합 토양이 Camelina sativa L.의 생장에 미치는 긍정적 영향)

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Lee, Hyeon-Sook;Sin, Jung-Ho;Kim, Eun-Suk;Woo, Hyo-Seop;Ahn, Sung-Ju
    • Ecology and Resilient Infrastructure
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    • v.5 no.3
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    • pp.163-173
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    • 2018
  • The biopolymer (BP) used in this study is mainly composed of xanthan gum and ${\beta}$-glucan derived from microorganism and has been introduced as a novel material for soil stabilization. However, the broad applicability of BP has been suggested in the field of geotechnical engineering while little information is available about the effects of BP on the vegetation. The goal of this study is to find the BP effects on the growth of Camelina sativa L. (Camelina) under drought condition. For more thorough evaluation of BP effects on the plant growth, we examined not only morphological but also physiological traits and gene expression patterns. After 25 days of drought treatment from germination in the soil amended with 0, 0.25, 0.5, and 1% BP, we observed that the BP concentration was strongly correlated the growth of Camelina. When plants were grown under drought stress, Camelina in 0.5% BP mixture showed better physiological parameters of the leaf stomatal conductance, electrolyte leakage and relative water content compared to those in control soil without BP. Plant recovery rate after re-watering was higher and the development of lateral root was lower in BP amended soil. RNA expression of Camelina leaf treated with/without drought for 7 and 10 days showed that aquaporin genes transporting solutes at bio-membrane, CsPIP1;4, 2;1, 2;6 and TIP1;2, 2;1, were induced more in the plants with BP amendment and drought treatment. These results suggest that the soil amended with BP has a positive effect on the transport of nutrients and waters into Camelina by improving water retention in soil under drought condition.