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검색결과 168건 처리시간 0.036초

뱀장어 근육내 Ryanodine Receptor의 기능 및 면역학적 성질 (Functional and Immunological Properties of Ryanodine Receptor in the Eel Skeletal Muscle)

  • 석정호;이연수;남장현;최숙정;홍장희;이재흔
    • 대한약리학회지
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    • 제31권2호
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    • pp.207-217
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    • 1995
  • To investigate the functional and immunological properties of the Ca-release channel in the sarcoplasmic reticulum(SR) of the eel skeletal muscle, $[^3H]ryanodine$ binding, SDS gel electrophoresis, $^{45}Ca\;release$ studies, and immunoblot assay were carried out in the SR of the eel skeletal muscle. Maximal binding sites(Bmax) and $K_D$ values of $[^3H]ryanodine$ for Ca-release channel of the SR of the eel skeletal muscle were $19.44{\pm}1.40\;pmole/mg$ protein and $15.55{\pm}1.69\;nM$, respectively. $[^3H]Ryanodine$ binding to RyR was increased by calcium and AMP. The SR of the eel skeletal muscle has two high molecular weight bands on the SDS PAGE. The mobility of upper band was more slower than the single band of the rabbit skeletal muscle, and that of the lower band was similar with the single band of canine cardiac muscle. Vesicular $^{45}Ca-release$ was activated by calcium. Ca-induced $^{45}Ca-release$ was significantly inhibited by $MgCl_2(2\;mM)$, ruthenium red$(10\;{/mu}M)$ or tetracaine(1 mM), but not by high concentration of calcium itself. AMP-induced $^{45}Ca-release$ was slightly occurred only in the absence of calcium, it was not inhibited by $MgCl_2$ or ruthenium red. Caffeine also increased $^{45}Ca-release$ from the SR vesicles, but it was not affected by $MgCl_2$ or ruthenium red. Polyclonal Ab against rat skeletal muscle RyR is reacted with that of rabbit, but not reacted with that of the eel skeletal muscle. These results suggested that ryanodine receptor of the SR of the eel skeletal muscle is showing some similar properties with that of mammalian skeletal muscle, but might be an another isotype channel having two bands which is less sensitive to AMP, not cross-reacted with antisera against rat RyR, and not inhibited by high concentration of calcium.

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LncRNA CRNDE Promotes the Progression of B-cell Precursor Acute Lymphoblastic Leukemia by Targeting the miR-345-5p/CREB Axis

  • Wang, Weimin;Wu, Feifei;Ma, Ping;Gan, Silin;Li, Xue;Chen, Li;Sun, Ling;Sun, Hui;Jiang, Zhongxing;Guo, Feng
    • Molecules and Cells
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    • 제43권8호
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    • pp.718-727
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    • 2020
  • The imbalance between the proliferation and apoptosis of B-cell precursors is an important contributor to the pathogenesis of B-cell precursor acute lymphoblastic leukemia (BCP-ALL), while its specific regulatory mechanism remains perplexing. This study aimed to expound the underlying mechanism of the proliferation and apoptosis of BCP-ALL cells from the perspective of non-coding RNA. In this study, long non-coding RNA colorectal neoplasia differentially expressed (LncRNA CRNDE) was upregulated in the bone marrow of BCP-ALL patients and BCP-ALL cell lines (NALM-6 and RS4;11). Functionally, LncRNA CRNDE knockdown restrained cell proliferation and boosted cell apoptosis in NALM-6 and RS4;11 cells. The subsequent investigation confirmed that LncRNA CRNDE bound to miR-345-5p and negatively regulated miR-345-5p expression. The overexpression of miR-345-5p suppressed cell proliferation and boosted cell apoptosis in NALM-6 and RS4;11 cells. Further experiments revealed that miR-345-5p downregulated cyclic AMP response element-binding protein (CREB) expression by targeting its mRNA directly. CREB overexpression reversed the effect of miR-345-5p mimic on cell proliferation and apoptosis in NALM-6 and RS4;11 cells. Finally, in vivo experiments showed that LncRNA CRNDE knockdown prolonged the survival of mice xenotransplanted with NALM-6 cells. In conclusion, LncRNA CRNDE upregulated CREB expression by suppressing miR-345-5p, thus promoting cell proliferation and reducing cell apoptosis in BCP-ALL.

3T3-L1 전구지방세포에서 개구리자리(Ranunculus sceleratus) 추출물의 AMPK 신호전달을 통한 지방생성 억제 효과 (Extract of Ranunculus sceleratus Reduced Adipogenesis by Inhibiting AMPK Pathway in 3T3-L1 Preadipocytes)

  • 김예지;조성필;이희주;홍금란;김경현;류시윤;정주영
    • 한방비만학회지
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    • 제22권1호
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    • pp.30-37
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    • 2022
  • Objectives: Adipogenesis is the process by which pre-adipocytes are differentiated into adipocytes. It also plays an important role in adipocyte formation and lipid accumulation. Ranunculus sceleratus (R. sceleratus) extracts are used for the treatment of various diseases such as hepatitis, jaundice, and tuberous lymphadenitis in oriental medicine. However, its effect on adipogenesis has not yet been studied. In this study, we investigated the effects of R. sceleratus on adipogenesis in 3T3-L1 cells. Methods: Cells were treated with 50, 100, and 200 ㎍/ml of R. sceleratus and cell viability was evaluated. To differentiate the 3T3-L1 preadipocytes, a 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI) solution were used. The accumulation of lipid droplets was determined by Oil Red O staining. The expression levels of adipogenesis-related proteins were also determined. Results: MDI solution differentiated the preadipocytes into adipocytes and accumulation of lipids was observed in the differentiated 3T3-L1 cells. Interestingly, the amount of lipid droplets was reduced after R. sceleratus treatment. In addition, the expression levels of key adipogenic transcription factors, such as CCAAT/enhancer-binding proteins-𝛼 (C/EBP-𝛼) and peroxisome proliferator-activated receptors-𝛾 (PPAR-𝛾) were also reduced after R. sceleratus treatment. Furthermore, R. sceleratus increased AMP-activated kinase (AMPK) phosphorylation and decreased sterol regulatory element-binding protein-1 expression. Conclusions: Our results showed that R. sceleratus reduced preadipocyte differentiation by inhibiting C/EBP-𝛼 and PPAR-𝛾 levels via the AMPK pathway. Therefore, we suggest that R. sceleratus may be potentially used as an anti-adipogenic agent.

Evaluation of DNA Extraction Methods from Low Copy Number (LCN) DNA Samples for Forensic DNA Typing

  • Eom, Yong-Bin
    • 대한의생명과학회지
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    • 제15권3호
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    • pp.229-232
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    • 2009
  • DNA isolation for PCR-based short tandem repeat (STR) analysis is essential to recover high yields of amplifiable DNA from low copy number (LCN) DNA samples. There are different methods developed for DNA extraction from the small bloodstain and gloves, commonly found at crime scenes. In order to obtain STR profiles from LCN DNA samples, DNA extraction protocols, namely the automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ method, the automated $QIAcube^{TM}$ method, the automated $Maxwell^{(R)}$ 16 DNA $IQ^{TM}$ Resin method, and the manual $QIAamp^{(R)}$ DNA Micro Kit method, were evaluated. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA Quantification Kit and DNA profiled by $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ Kit. Results were compared based on the amount of DNA obtained and the completeness of the STR profiles produced. The automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ and $QIAcube^{TM}$ methoas produced reproducible DNA of sufficient quantity and quality trom the dried blood spot. This two automated methods showed a quantity and quality comparable to those of the forensic manual standard protocols normally used in our laboratory. In our hands, the automated DNA extraction method is another obvious choice when the forensic case sample available is bloodstain. The findings of this study indicate that the manual simple modified $QIAamp^{(R)}$ DNA Micro Kit method is best method to recover high yields of amplifiable DNA from the numerous potential sources of LCN DNA samples.

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Antiobesity effects of the water-soluble fraction of the ethanol extract of Smilax china L. leaf in 3T3-L1 adipocytes

  • Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Dae Jung;Choe, Myeon
    • Nutrition Research and Practice
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    • 제9권6호
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    • pp.606-612
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    • 2015
  • BACKGROUND/OBJECTIVES: Several medicinal properties of Smilax china L. have been studied including antioxidant, anti-inflammatory, and anti-cancer effects. However, the antiobesity activity and mechanism by which the water-soluble fraction of this plant mediates its effects are not clear. In the present study, we investigated the lipolytic actions of the water-soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) in 3T3-L1 adipocytes. MATERIALS/METHODS: The wsSCLE was identified by measuring the total polyphenol and flavonoid content. The wsSCLE was evaluated for its effects on cell viability, lipid accumulation, glycerol, and cyclic adenosine monophosphate (cAMP) contents. In addition, western blot analysis was used to evaluate the effects on protein kinase A (PKA), PKA substrates (PKAs), and hormone-sensitive lipase (HSL). For the lipid accumulation assay, 3T3-L1 adipocytes were treated with different doses of wsSCLE for 9 days starting 2 days post-confluence. In other cell experiments, mature 3T3-L1 adipocytes were treated for 24 h with wsSCLE. RESULTS: Results showed that treatment with wsSCLE at 0.05, 0.1, and 0.25 mg/mL had no effect on cell morphology and viability. Without evidence of toxicity, wsSCLE treatment decreased lipid accumulation compared with the untreated adipocyte controls as shown by the lower absorbance of Oil Red O stain. The wsSCLE significantly induced glycerol release and cAMP production in mature 3T3-L1 cells. Furthermore, protein levels of phosphorylated PKA, PKAs, and HSL significantly increased following wsSCLE treatment. CONCLUSION: These results demonstrate that the potential antiobesity activity of wsSCLE is at least in part due to the stimulation of cAMP-PKA-HSL signaling. In addition, the wsSCLE-stimulated lipolysis induced by the signaling is mediated via activation of the ${\beta}$-adrenergic receptor.

우즈베키스탄 Maidanak 천문대 Fairchild 486 CCD의 기본적인 특성 (CHARACTERISTICS OF THE FAIRCHILD 486 CCD AT MAIDANAK ASTRONOMICAL OBSERVATORY IN UZBEKISTAN)

  • 임범두;성환경
    • 천문학논총
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    • 제23권1호
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    • pp.1-12
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    • 2008
  • Understanding of the basic characteristics of an astronomical instrument is a prerequisite to obtaining reliable data from the instrument. We have analyzed more than 1,000 calibration images from the Fairchild 486 CCD (hereafter the Maidanak 4k CCD system) attached to the AZT-22 1.5m telescope at Maidanak Astronomical Observatory in Uzbekistan. The Maidanak 4k CCD system supports three readout modes through 1, 2, or 4 amplifiers. In most cases observers use 4-amplifier readout mode to save time. We have tested the stability and seasonal variation of zero levels and confirm that two quadrants of the images (Amp 1 & 2) show no appreciable seasonal variation. but the other two quadrants (Amp 3 & Amp 4) show an evident seasonal variation in the bias level. The Cryo Tiger, the cooling system used at the Maidanak 4k CCD system, maintains the CCD temperature at -108'E, and effectively suppresses the dark electrons. The mean value versus the variance plot of the flat images does not show the expected relation for an ideal Poisson noise distribution and this is attributed to the large variation in quantum efficiency between different pixels. In addition, we confirm that there is no appreciable difference in gain between readout amplifiers, but there is a large variation in quantum efficiency across CCD chip especially in U. Due to the finite length of shutter opening and closing time, the effective exposure time varies across the science images. We introduce two parameters to quantify the effect of this uneven illumination and present a method to remove these effects. We also present a method to remove the interference patterns appearing in the images obtained with longer wavelength filters and investigate the spatial variation of the point spread function.

Clamping Op-Amp를 이용한 광대역 로그 비디오 증폭기의 선형성 개선에 관한 연구 (Study on the Development of Linearity of Broad-Band SDLVA Using Clamping Op-Amp)

  • 박종설;김종건;김점식
    • 한국전자파학회논문지
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    • 제22권6호
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    • pp.641-647
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    • 2011
  • 본 논문은 0.5~2.0 GHz에서 -70~0 dBm의 동적 범위를 갖는 로그 비디오 증폭기(Successive Detecting Log Video Amplifier, 이하 SDLVA)를 설계 및 제작하였다. 제작된 SDLVA는 5단의 증폭기를 이용한 고주파 출력부, 2단의 로그검파기 그리고 선형성 개선을 위하여 클램핑 연산증폭기를 이용한 가산 회로로 구현하였다. 제작된 SDLVA의 측정 결과는 73 dB 이상의 소신호 이득과 10.1~12.2 dBm의 고주파 포화 출력 특성을 갖는다. 그리고 -70~0 dBm 입력 시 25 mV/ dB${\pm}$1.0 mV의 기울기를 갖고 ${\pm}$1.5 dB 이하의 선형 특성을 갖는 비디오 신호를 출력한다.

Anti-thrombotic effects of ginsenoside Rk3 by regulating cAMP and PI3K/MAPK pathway on human platelets

  • Hyuk-Woo Kwon ;Jung-Hae Shin ;Man Hee Rhee ;Chang-Eun Park ;Dong-Ha Lee
    • Journal of Ginseng Research
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    • 제47권6호
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    • pp.706-713
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    • 2023
  • Background and objective: The ability to inhibit aggregation has been demonstrated with synthetically derived ginsenoside compounds G-Rp (1, 3, and 4) and ginsenosides naturally found in Panax ginseng 20(S)-Rg3, Rg6, F4, and Ro. Among these compounds, Rk3 (G-Rk3) from Panax ginseng needs to be further explored in order to reveal the mechanisms of action during inhibition. Methodology: Our study focused to investigate the action of G-Rk3 on agonist-stimulated human platelet aggregation, inhibition of platelet signaling molecules such as fibrinogen binding with integrin αIIbβ3 using flow cytometry, intracellular calcium mobilization, dense granule secretion, and thromboxane B2 secretion. In addition, we checked the regulation of phosphorylation on PI3K/MAPK pathway, and thrombin-induced clot retraction was also observed in platelets rich plasma. Key Results: G-Rk3 significantly increased amounts of cyclic adenosine monophosphate (cAMP) and led to significant phosphorylation of cAMP-dependent kinase substrates vasodilator-stimulated phosphoprotein (VASP) and inositol 1,4,5-trisphosphate receptor (IP3R). In the presence of G-Rk3, dense tubular system Ca2+ was inhibited, and platelet activity was lowered by inactivating the integrin αIIb/β3 and reducing the binding of fibrinogen. Furthermore, the effect of G-Rk3 extended to the inhibition of MAPK and PI3K/Akt phosphorylation resulting in the reduced secretion of intracellular granules and reduced production of TXA2. Lastly, G-Rk3 inhibited platelet aggregation and thrombus formation via fibrin clot. Conclusions and implications: These results suggest that when dealing with cardiovascular diseases brought upon by faulty aggregation among platelets or through the formation of a thrombus, the G-Rk3 compound can play a role as an effective prophylactic or therapeutic agent.

채종원관리도로 성력화를 위한 연구 -친환경적 횡단목배수로의 적용 가능성에 대한 검토-

  • 이성기;김종한;이갑연;이성규;김태구
    • 한국환경과학회:학술대회논문집
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    • 한국환경과학회 2004년도 봄 학술발표회 발표논문집
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    • pp.137-140
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    • 2004
  • 채종원관리를 효율적으로 관리하기 위하여 시설되는 저 규격도로는 대부분 비포장상태로 이용되는 경우 많다. 이는 강우에 의한 침식붕괴로 하류의 산림식생환경 및 수질환경 오염으로 연결될 위험성이 내포되어 있어 적절한 방법으로 사전에 방지할 필요가 있다. 본 연구에서는 환경과 친화적인 재료를 최대한 이용하여 개발된 목배수로를 내구성 강화를 검토하였던바 섬유질 포의 조절과 고무질의 성능 조절에 의한 검토로 현장에의 적용이 가능하였다.

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고콜레스테롤 식이 섭취 쥐에서 quercetin의 간 AMPK 및 microRNA-21 조절을 통한 지질대사 개선 효과 (Effects of quercetin on the improvement of lipid metabolism through regulating hepatic AMPK and microRNA-21 in high cholesterol diet-fed mice)

  • 이막순;김양하
    • Journal of Nutrition and Health
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    • 제55권1호
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    • pp.36-46
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    • 2022
  • Quercetin의 지질대사 개선 효과에 대한 작용기전을 확인하기 위해 C57BL/6J mouse를 사용하여 실험을 수행하였다. 고콜레스테롤혈증을 유도하기 위해 6주간 1% 콜레스테롤과 0.5% cholic acid를 함유하는 고콜레스테롤 식이를 급여하였으며, quercetin은 0.05%와 0.1%의 수준으로 고콜레스테롤 식이에 추가하여 같은 기간 동안 제공하였다. Quercetin은 혈청과 간의 중성지방 및 콜레스테롤 수준을 용량 의존적으로 감소하는 것으로 나타났다. 고콜레스테롤 식이를 섭취한 쥐의 간에서 지방 합성을 촉진하는 SREBP-1c, ACC1 및 FAS 유전자 발현이 quercetin 섭취에 의해 억제되는 것을 확인하였다. Quercetin은 간세포 내에서 에너지 대사를 조절하는 AMPK 활성을 증가시켰다. 이에 반해 암세포 증식을 촉진하고 지방간에서 높게 발현되는 miR-21 발현은 quercetin 섭취에 의해 감소되었다. 본 연구의 결과는 quercetin이 고콜레스테롤 식이 섭취 쥐에서 혈청과 간의 지질 수준을 낮추는 지질대사 개선 효과가 있으며, 이러한 효과의 일부는 간 내 지방합성 유전자 (SREBP-1c, ACC1 및 FAS) 발현, AMPK 활성 및 miR-21 조절을 통해 매개된다는 것을 시사한다.