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한우 난소 과립막세포의 체외배양에서 Follicular Fluid 및 Anti-Inhibin Serum의 첨가효과

  • 성환후;최선호;장유민;박진기;장원경;정일정
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.33-33
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    • 2001
  • 본 연구는 한우 난포발달에 있어서 난포액 및 inhibin의 생리적 역할을 검토하기 위해 수행하였다. Anti-inhibin serum(AIS) 생산을 위해 사용된 항원은 porcine inhibin-$\alpha$-subunit 19~32의 peptide를 사용하여 adjuvant 용액을 1:3의 비율로 혼합하여 앙고라종 토끼 5두(체중 2.5kg)에게 주 2회 간격으로 8회 실시후 ELISA Leader로 항혈청의 역가를 확인하였다. 난포액(bFF; bovine follicular fluid)은 도축장에서 도축되는 한우 난소로부터 직경 1.0cm 이하의 난포로부터 회수하여 스테로이드를 제거하기 위해 10% chacoal solution(50 mg/$m\ell$, Norrit-A, Fisher Sci., USA)을 처리하여 45분간 배양후 원심분리후 상층액을 회수하여 실험에 공시하였다. 과립막세포의 체외배양을 위해 D-MEM용액(10% FCS와 antibiotics를 첨가)을 배양액으로 하여 1 $\times$ $10^{6}$ cells/$m\ell$로 조절하였다. 호르몬은 RIA 및 ELISA법으로 분석하였다. 그 결과 항원-항체반응은 항원처리후 24일째부터 항체가를 확인할 수 있었으며 52일째에서 높은 항원-항체반응을 보였다. 한편, 난포크기별 난포액의 progesterone 및 Estradiol-l7$\beta$을 농도를 분석한 결과, Progesterone 난포크기가 직경 1.0 cm부터 유의적으로 증가하기 시작하여 직경 2.0cm의 난포액에서는 높은 progesterone이 존재하고 있는 것으로 나타났다. 이에 반해, 난포크기별 난포액중 estradiol 17$\beta$농도는 직경 1.0cm구에서 가장 높게 나타났다. 난포직경이 1cm일 경우에 난포액내 etradiol-17$\beta$가 가장 많이 존재하고 있음이 나타났다. 과립막세포의 체외배양에서 배양 24시간에서는 과립막세포의 progesterone분비는 약 40ng/1$\times$$10^{6}$ cell/well/$m\ell$ 전후로 나타났으며 bFF 5%, AI 5% 및 bFF+AI 첨가구에 따라 유의적인 차이는 없었다 반면에 48시간배양구에서는 24시간에 비해 유의적으로 높게 분비되었으며, bFF 5%처리구와 bFF5%+AI5%처리구에서는 progesterone을 대조구보다 유의적으로 억제되었으나 AI 5%단독처리구에서는 대조구와 큰 차이가 없었다. 한편, 각 세포질을 SDS-PAGE로 분리하여 nitro cellulose membrane에 transfer하여 Western blotting법에 의해 검토한 결과, 직경 1.0 cm의 성숙난포의 granulosa cell에 특이하게 Inhibin이 존재하고 있음이 확인되었다. 이상의 결과로, 한우에 있어서 성숙난포에 존재하는 Inhibin은 난포발달 및 난포세포의 스테로이드호르몬합성에 중요하게 관여하고 있는 것으로 사료된다.

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옻나무 유래 Flavonoid처리가 흰쥐 Leydig 세포의 체외배양에서 Testosterone분비에 미치는 영향

  • 성환후;최선호;민관식;장유민;나천수;정일정
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.80-80
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    • 2001
  • 옻나무 유래 Flavonoid(F)는 항암 효과, 숙취해소 등의 효능이 있으며, 신장의 기능을 촉진하고 생식에도 영향을 준다고 알려져 있다. 그 주성분으로는 fustin, fisetin, sulfuretin, butein으로 추정되고 있다. 본 연구팀의 예비실험에서 성숙 흰쥐에게 F를 30일간 5mg씩 구강투여 하여 정소 중량 및 혈액 내 Testosterone 농도를 분석한 결과, 정소 중량이 증가되고 혈중 Testosterone의 농도도 대조구에 비해 유의적으로 높은 결과를 얻었다. 이에 본 연구의 목적은 옻나무 유래 F가 성숙 수컷 횐쥐의 생식 기능에 직접 미치는 영향을 알아보고자, 흰쥐 Leydig 세포를 분리, 회수하여 체외배양에서 Testosterone농도를 조사하였다. SD계 흰쥐(약 12주령, 체중 250g 전후)의 정소를 적출 한 후, Leydig 세포를 회수하기 위해 0.25% collagenase용액에 넣어 34$^{\circ}C$에서 15분간 진탕 수조에서 배양하였다. 배양된 정소조직은 D-MEM 배양액(10% FCS와 antibiotics 첨가)으로 2~3회 세척한 후 세포 수를 1$\times$$10^{6}$ cells/$m\ell$/well로 분주하여, F(20, 40, 80, 160ng), IGF-I(50, 100ng) 와 LH(10, 100ng) 용량별 각각 첨가하여 배양하였다. 각 처리 후, 배양시간(3, 6, 12, 24, 36시간)에 따라 배양액을 회수하여 COAT-A-COUNT(DPC, USA) kit를 이용하여 RIA방법에 의해 Testosterone을 분석한 결과는 다음과 같다. 대조구 Leydig 세포를 36시간까지 체외 배양하여 Testosterone의 농도를 조사한 결과, 24시간 배양구가 가장 높은 농도를 나타내었다. F를 단독 첨가하여 12시간 배양한 실험에서 F 80ng 첨가구에서 유의적으로 높은 농도를 보였다. LH 10ng 및 100ng 첨가구의 시간별 변화로는, LH 10ng 첨가구에서는 6~12시간 이후, LH 100ng 첨가구에서는 3~6시간 사이에서 유의적인 증가를 보였다. 또한, LH 10ng 첨가 실험에서는 LH 10ng+F 40ng에서 12시간 배양 시, 유의적으로 높은 Testosterone 분비를 확인하였으며, LH 100ng을 첨가하여 3시간 배양 시는 각 처리구 마다 높은 유의적 차이를 보였다. 한편, GH에 관여하는 IGF-I 첨가효과를 비교 검토한 결과, LH 100ng+IGF-I 100ng, 6시간 배양구에서 유의적으로 높게 나타내었다. 이상의 결과로, F 단독 처리 시의 적정량은 약 80ng이며, 흰쥐 정소 Leydig 세포의 Testosterone분비를 촉진하는 작용을 하는 것으로 사료되며, 특히 LH+F구에서 Testosterone분비를 더욱 향상시킴으로써 정소 Leydig 세포의 Androgen 생성을 촉진시키는 역할이 있는 것으로 보여진다 따라서 옻나무 유래 F는 포유동물의 생식기능에 중요하게 작용하는 것으로 사료된다.

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Roles of Neutrophil/Lymphocyte and Platelet/Lymphocyte Ratios in the Early Diagnosis of Malignant Ovarian Masses

  • Yildirim, Mem Arjen;Seckin, Kerem Doga;Togrul, Cihan;Baser, Eralp;Karsli, Mehmet Fatih;Gungor, Tayfun;Gulerman, Hacer Cavidan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6881-6885
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    • 2014
  • Background: The present study aimed to investigate the utility and importance of the various parameters of complete blood count panel for benign-malignant differentiation of adnexal masses. Materials and Methods: This retrospective study involved 316 patients with documented benign and 253 patients with malignant adnexal masses who underwent primary surgical treatment at a tertiary referral center. Prior to the study, all benign and malignant cases were compared within their own groups and then the benign and malignant cases were compared to each other. For all cases, cut-off, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated for the neutrophil lymphocyte ratio (NLR), platelet lymphocyte ratio (PLR), neutrophil, lymphocyte, platelet and CA-125 parameters, and the results were compared in regards to the groups. Results: NLR, PLR, neutrophil, CA-125, and platelet values were higher in the malignant compared to the benign cases (p<0.01). The lymphocyte value was lower in the malignant cases (p<0.01). No significant differences were found for basophils and eosinophils (p > 0.05). For CA-125, the sensitivity, specificity, PPV and NPV for all cases were 78%, 62%, 62% and 78%, respectively. For NLR, they were 65.6%, 72.1%, 65.3%, and 72.3%, and for PLR, 48%, 81%, 67%, and 66%. Additionally, the sensitivity and specificity were 55% and 77% for CA-125, 66% and 58% for NLR, and 61% and 58% for PLR in early malignant cases. Conclusions: NLR and PLR appear to be useful methods that can be applied together with CA-125 due to the relatively high sensitivity values for the malign-benign differentiation of ovarian masses. Although the specificity of these parameters is lower than CA-125, especially in cases with early malignant ovarian pathology, their sensitivity being higher is promising for the early diagnosis of ovarian cancer. It can be used to detect ovarian malignancies in the early stages, and it will increase the treatment options and improve survival rates.

A study on the characteristics of MEM structure of $SrBi_2Ta_2O_9$ thin films by RE magnetron sputtering (RF 마그네트론 스퍼터링법에 의한 MFM 구조의 $SrBi_2Ta_2O_9$ 박막 특성에 관한 연구)

  • 이후용;최훈상;최인훈
    • Journal of the Korean Vacuum Society
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    • v.9 no.2
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    • pp.136-143
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    • 2000
  • $SrBi_2Ta_2O_9;(SBT)$ films were deposited on p-type Si(100) at room temperature by rf magnetron sputtering method to confirm the possibility of application of $Pt/SBT/Pt/Ti/SiO_2/Si$ structure (MFM) for destructive read out ferroelectric RAM (random access memory). Their structural characteristics with the various annealing times and Ar/$O_2$ gas flow ratios in sputtering were observed by XRD (X-ray diffractometer) and the surface morphologies were observed by FE-SEM (field emission scanning electron microscopy), and their electrical properties were observed by P-V (polarization-voltage measurement) and I-V (current-voltage measurement). The Ar/$O_2$ gas flow ratios of sputtering gas were changed from 1 : 4 to 4 : 1 and SBT thin films were deposited at room temperature. The films show (105), (110) peaks of SBT by XRD measurement. SBT thin films deposited at room temperature were crystallized by furnace annealing at 80$0^{\circ}C$ in oxygen atmosphere during either one hour or two hours. Among their electrical properties, P-V curves showed shaped hysteresis curves, but the SBT thin films showed the asymmetric ferroelectric properties in P-V curves. When Ar/$O_2$ gas flow ratios are 1 : 1, 2: 1, the leakage current density values of SBT thin films are good, those values of 3 V, 5 V, and 7 V are respectively $3.11\times10^{-8} \textrm{A/cm}^2$, $5\times10^{-8}\textrm{A/cm}^2$, $7\times10^{-8}\textrm{A/cm}^2$.After two hours of annealing time, their electrical properties and crystallization are improved.

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An Improvement of Speed for Wavelength Multiplex Optical Network using Optical Micro Electro Mechanical Switches (광마이크로전자기계 스위치를 이용한 파장다중 광네트워크의 속도 재선)

  • Lee Sang-Wha;Song Hae-Sang
    • Journal of the Korea Society of Computer and Information
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    • v.10 no.5 s.37
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    • pp.123-132
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    • 2005
  • In this Paper, we present an improvement of switch node for wavelength multiplex optical network. Currently because of quick increase of internet traffic a big network capacity is demanded. Wavelength multiplex optical network Provides the data transfer of high speed and the transparent characteristic of the data. Therefore optic network configuration is the most powerful technology in the future. It will be able to control the massive traffic from the optical network in order to transmit the multimedia information of very many quantify. Consequently the node where the traffic control is Possible, is demanded. The optical switch node which manages efficiently the multiple wavelength was Proposed. This switch is composed of a optical switch module for switching and a wavelength converter module for wavelength conversion. It will be able to compose the switch fabric without optical/electro or electro/optical conversion using optical MEMS(Micro Electro Mechanical Switches) module. Finally, we present the good test result regarding the operational qualify of the switch fabric and the performance of optical signal from the switch node. The proposed switch node of the optic network will be able to control the massive traffic with all optical.

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Effects of Extracts of Natural Products on Alkaline Phosphatase Activity of MC3T3 - E1 Cells (수종의 생약추출물이 MC3T3-E1 세포의 염기성 인산분해 효소 활성에 미치는 영향)

  • Park, Sang-Kee;Kim, Dae-Kyum;You, Seung-Han;Kim, Hyun-A;Kim, Myoung-Dong;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.123-135
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    • 2001
  • Several growth factors and polypeptides were studied for the regeneration of periodontal supporting tissues which had been lost due to periodontal disease. But these are not commonly used for regenerators of bone tissue or alveolar bone, because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many natural products, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential or periodontal tissues. Cnidii Rhizoma, Rhinocerotis Cornu and Drynariae Rhizoma have been traditionally used as a drug for treatment of bone disease in oriental medicine. The purpose of this study was to examine the ability of alkaline phosphatase synthesis of MC3T3-E1 cells when above medicines were supplimented. MC3T3-E1 cells were cultured with ${\alpha}-MEM(negative control)$, dexamethasone(positive control), and each natural products for 3 and 5 days. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry. Except Cnidii Rhizoma, all of the natural products of this study induced higher activity of ALP synthesis than controls. Among them Drynariae Rhizoma induced the highest activity. In the aspects of culturing time, all medicines did not showed the difference between 3 and 5 days, but $10^{-7}g/ml$ group of Rhinocerotis Corun showed significant increase at 3 days than at 5 days. These results indicate that several natural products have a inducing ability of ALP synthesis on osteoblasts.

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A High Yield Rate MEMS Gyroscope with a Packaged SiOG Process (SiOG 공정을 이용한 고 신뢰성 MEMS 자이로스코프)

  • Lee Moon Chul;Kang Seok Jin;Jung Kyu Dong;Choa Sung-Hoon;Cho Yang Chul
    • Journal of the Microelectronics and Packaging Society
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    • v.12 no.3 s.36
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    • pp.187-196
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    • 2005
  • MEMS devices such as a vibratory gyroscope often suffer from a lower yield rate due to fabrication errors and the external stress. In the decoupled vibratory gyroscope, the main factor that determines the yield rate is the frequency difference between the sensing and driving modes. The gyroscope, fabricated with SOI (Silicon-On-Insulator) wafer and packaged using the anodic bonding, has a large wafer bowing caused by thermal expansion mismatch as well as non-uniform surfaces of the structures caused by the notching effect. These effects result in large distribution in the frequency difference, and thereby a lower yield rate. To improve the yield rate we propose a packaged SiOG (Silicon On Glass) technology. It uses a silicon wafer and two glass wafers to minimize the wafer bowing and a metallic membrane to avoid the notching. In the packaged SiOG gyroscope, the notching effect is eliminated and the warpage of the wafer is greatly reduced. Consequently the frequency difference is more uniformly distributed and its variation is greatly improved. Therefore we can achieve a more robust vibratory MEMS gyroscope with a higher yield rate.

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Effect of Glucose at High Concentrations on the Integrin and Cathepsin Expression of the Cultured Human Periodontal Ligament Cells (고농도 포도당이 사람 치주인대세포의 Integrin과 Cathepsin 발현에 미치는 영향에 관한 연구)

  • Kim, Bang-Soo;Shin, Je-Won;Hong, Jung-Pyo;Jue, Seong-Suk
    • Journal of Oral Medicine and Pain
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    • v.33 no.1
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    • pp.35-40
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    • 2008
  • This experiment was designed to clarify the effect of high concentrations of extracellular glucose on the periodontal ligament cells. The cells were incubated for 24 and 48 hours with ${\alpha}-MEM$ including 1,000 mg/L(control group) and 4,500 mg/L(experimental group) of glucose. Then, the expressions of ${\alpha}5$ integrin, cathepsin-B and -L were examined using RT-PCR method. The results were as follows: 1. ${\alpha}5$ integrin expression was increased after incubation in high glucose media. 2. Cathepsin-B expression was increased after the 24-H incubation with high glucose media, but was decreased after the 48-H incubation. 3. The expression of cathepsin-L was decreased by the high glucose media. Theses results suggest that extracellular glucose at high concentration may be attributed to delayed wound healing in diabetes patients through increase in ${\alpha}5$ integrin and decrease in cathepsins, which lead to accumulation of extracellular matrices and adhesion molecules.

Effects of 2-deoxy-D-glucose and quercetin on osteoblastic differentiation and mineralization in irradiated MC3T3-E1 cells (2-deoxy-D-glucose와 quercetin이 방사선조사 MC3T3-E1 골모세포주의 분화와 석회화에 미치는 영향)

  • Ahn, Hyoun-Suk;Kim, Kyoung-A;Koh, Kwang-Joon
    • Imaging Science in Dentistry
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    • v.36 no.4
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    • pp.189-198
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    • 2006
  • Purpose: To investigate the in vitro response of MC3T3-E1 osteoblastic cells to X-ray in the presence and absence of 2 deoxy-D-glucose (2-DG) and quercetin (QCT). Materials and Methods: The MC3T3-E1 cells were cultured in an ${\alpha}-MEM$ supplemented with 5 mM 2-DG or $10{\mu}M$ QCT and then the cells were incubated for 12 h prior to irradiation with 2, 4, 6, and 8Gy using a linear accelerator (Mevaprimus, Germany) delivered at a rate of 1.5 Gy/min. At various times after the irradiation, the cells were processed for the analyses of proliferation, viability, cytotoxicity, and mineralization. Results: Exposure of the cells to X-ray inhibited the tritium incorporation, 3-(4, 5-dimethylthiazol-2yl-)-2, 5-diphenyl tetrazolium bromide (MTT)-reducing activity, and alkaline phosphatase (ALP) activity, and caused cytotoxicity and apoptosis in a dose-dependent manner of the X-ray. This effect was further apparent on day 3 and 7 after the irradiation. RA+2-DG showed the decrease of DNA content, cell viability, and increase of cytotoxicity rather than RA. ALP activity increased on day 7 and subsequently its activity dropped to a lower level. 2-DG suppressed the calcium concentration, but visual difference of number of calcified nodules between RA and RA+2-DG was not noticed. RA+QCT showed the increase of DNA content, cell viability, but decrease of cytotoxicity and subG1 stage cells in the cell cycle, and increased calcified nodules in von Kossa staining rather than the RA. ALP activity showed significant increases on day 7 and subsequently its activity dropped to a lower level. Conclusion: The results showed that the 2-DG acted as a radiosensitizing agent and QCT acted as a radiosensitizing agent respectively in the irradiated MC3T3-E1 osteoblast-like cells.

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A STUDY OF THE EFFECT OF ZIZYPHUS FRUCTUS EXTRACTS ON MORPHOLOGY & CHEMOTAXIS OF GINGIVAL FIBROBLAST & PERIODONTAL LIGAMENT CELLS (치은섬유아세포와 치은인대세포의 형태와 화학주성에 미치는 대조추출물의 효과에 관한 연구)

  • Yang, Chang-Ho;Ryu, In-Chul;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.279-289
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    • 1995
  • The most important object of periodontal treatment is the perfect regeneration of destructed periodontal tissue. The healing of periodontal lesion is affected by several cells & factors, which result in formation of long juntional epithelium, root resorption, bony ankylosis or connective tissue attachment. And ideal healing is enhanced by epithilial exclusion or periodontal ligament cell activation. In this investigation, I studied the effect of Zizyphus Fructus extract which enhances biologic activity& collagen synthesis, on the chemotaxis & cell nature. The cells were obtained from interdental area & middle third area of the freshly extracted teeth for the orthodontic purpose. And they were fully incubated in${\alpha}-MEM$ solution containing $100{\mu]g/ml$ penicillin & $100{\mu]g/ml$ streptomycin followed by 6 generation incubation. The test cells were collected by trypsin-EDTA & centrifuge in the fully incubated cells, counted by Hernacyotmeter, incbated $5{\times}10^5/ml$ cells for 24 hours, re-incubated 24 hours in media containing natural extract and photographed. The cells were incubated for 4 hours in 48 well microchemotaxis chamber bisecting upper & lower chamber by 8ug/m pore polycarbonate membrane coating 5mg/ml gelatin solution. The migrated cells in microscope were counted, which meaned cell chemotaxis activity. The study had shown that the morphology of cell was spindle-shaped as the control group, and the subextract test groups were not significantly different. In gingival fibroblasts, the chemotaxis effect of PDGF was statistically significant compared to control group. The Zizyphus Fructus extract was more or less enhanced chemotaxis effect and in $1{\mu}g/ml$ concentration the chemotaxis effect was slightly elevated compared with $10{\mu}g/ml$ concentration. But, among the subextracts, it was not significantly defferent. In PDL cells, the chemotaxis effect of PDGF in statistically significant, and the zizyphus Fructus extract had shown the enhanced effect. The effect was slightly higher in $1{\mu}g/ml$ concentration than 10g/ml concentration,and no significance among the subextracts.

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