• 제목/요약/키워드: Serum DNA

검색결과 409건 처리시간 0.026초

골아세포의 IGF-I 유전자 발현 및 세포증식에 대한 1,25-dihydroxyvitamin $D_3$의 영향 (The Effects of 1,25- Dihydroxyvitamin $D_3$ on Expression of IGF-I Gene and Cellular Proliferation in MC3T3-E1 Cells)

  • 최희동;이재목;서조영
    • Journal of Periodontal and Implant Science
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    • 제30권1호
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    • pp.39-52
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    • 2000
  • Polypeptide growth factor belong to a class of potent biologic mediator which regulate cell differentiation, proliferation, migration and metabolism. 1,25-dihydroxyvitamin $D_3$ decrease cell proliferation, and stimulate alkaline phosphatase activity which express in osteoblast during cell differentiation period. IGF-I is known to stimulate cell proliferation and differentiation too. 1,25-dihydroxyvitamin $D_3$ is known to increase IGF-I binding sites and IGF binding protein which inhibite the effect of IGF. The purpose of this study is to evaluate potential role of IGF-I as mediator that control the action of 1,25-dihydroxyvitamin $D_3$. MC3T3-E1 cell were seeded $5{\times}10^5/ml$ at 100mm culture plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 5% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ added. Total mRNA was extracted at 0, 6, 24, 48, 72 hour. PRPCR method was programed for the detection of IGF-I mRNA. In the both groups of 1,25-dihydroxy vitamin $D_3$ treated and control, alternative splicing form of IGF-I, IGF-IA and IGF-IB were expressed. In the 1,25-dihydroxyvitamin $D_3$ treated group, IGF-I mRNA expression was matained until 24 hour, there after expression was decresed. MC3T3-E1 cell were seeded $2.5{\times}10^4/ml$ at 24well plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 3% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ and 10 ng/ml IGF-I were added separately or together. Cell were cultured for 1 and 3 days, $2{\mu}Ci/ml\;[^3H]$ -thymidine was added for the last 24h of culture of each days. ${[^3H]}$-thymidine incorporation in to DNA was measured and expressed counter per minute(CPM). DNA synthetic activity was significantly decreased by 1,25-dihydroxyvitamin $D_3$ both at 1 day and 3 day, and in the combination group of 1,25-dihydroxyvitamin $D_3$ and IGF-I, DNA synthetic activity was also decreased both at 1 day and 3 days. IGF-I did not affect the DNA synthetic activity compared to control group both at 1 day and 3 day. From the above results, 1,25-dihydroxyvitamin $D_3$ was potent inhibitor of cell proliferaton in MC3T3-E1 cells. It assumed that the effect of 1,25-dihydroxyvitamin $D_3$ on osteoblast proliferation may be mediated in part by decreased level of IGF-I.

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Advantages of the single nucleotide polymorphism-based noninvasive prenatal test

  • Kim, Kunwoo
    • Journal of Genetic Medicine
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    • 제12권2호
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    • pp.66-71
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    • 2015
  • Down syndrome screening with cell-free DNA (cfDNA) in the maternal plasma has recently received much attention in the prenatal diagnostic field. Indeed, a large amount of evidence has already accumulated to show that screening tests with cfDNA are more sensitive and specific than conventional maternal serum and/or ultrasound screening. Globally, more than 1,000,000 of these noninvasive prenatal tests (NIPTs) have been performed to date. There are several different methods for NIPTs that are currently commercially available, including shotgun massively parallel sequencing, targeted massively parallel sequencing, and single nucleotide polymorphism (SNP)-based methods. All of these methods have their own advantages and disadvantages. In this review, I will focus specifically on the SNP-based NIPT.

The Effects of Fetal Bovine Serum, Epidermal Growth Factor, and Retinoic Acid on Adult Rat Islets Embedded in Collagen Gels

  • Shin, Jun-Seop;Chang, Hyo-Ihl;Sung, Ha-Chin;Kim, Chan-Wha
    • Journal of Microbiology and Biotechnology
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    • 제9권2호
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    • pp.150-156
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    • 1999
  • The induction of proliferation of adult rat islets was investigated under various culture conditions. The islets were isolated from male Sprague-Dawley rats and subsequently embedded in collagen gels, which mimic the in vivo three-dimensional surroundings. During the culture period, the effects of heterologous serum (fetal bovine serum, FBS), epidermal growth factor (EGF), and retinoic acid (RA) on islet growth were examined with respect to the morphological and total DNA content changes. To investigate these changes at the cellular level, whole mount immunocytochemistry using specific antibodies for insulin and glucagon was performed. The results showed that (i) collagen gels as an extracellular matrix can maintain islets in a similar way to that in vivo, (ii) heterologous serum (FBS) had stimulatory effects on islet proliferation in a dose-dependent manner, (iii) RA had inhibitory effects on islet proliferation induced by the serum in a dose-dependent manner, (iv) EGF had weak inhibitory effects on islet proliferation induced by the serum except at the concentration of 10 nM where its effect was not significant, and (v) whole mount immunocytochemistry revealed that newly proliferated islet cells were mainly $\beta$-and $\alpha$-cells.

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The Serum Concentrations of YKL-40, IL-6, and TNF-${\alpha}$ in Retired Workers Exposed to Inorganic Dusts

  • Lee, Kyung Myung;Shin, Jae Hoon;Hwang, JooHwan;Lee, Jong Seong;Choi, Byung-Soon;Kim, In Sik
    • 대한의생명과학회지
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    • 제20권1호
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    • pp.8-13
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    • 2014
  • Occupational long-term exposure to inorganic dusts may cause a variety of lung diseases such as pneumoconiosis and chronic obstructive pulmonary disease (COPD). Diagnosis of pneumoconiosis and COPD, however, is currently dependent on radiological findings and pulmonary test, which are both late diagnostic tools. Therefore, there is a need to identify novel biomarkers in pneumoconiosis and COPD. Hence, in this current study we investigated the serum concentrations of YKL-40, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-${\alpha}$) as biomarkers for pneumoconiosis and COPD in 161 retired male workers exposed to inorganic dusts. The serum concentration of YKL-40 was significantly increased with age, pneumoconiosis, and airflow limitation. The serum concentration of IL-6 was significantly higher in airflow limitation. These results suggest that serum concentration of YKL-40 is associated with age, pneumoconiosis, and airflow limitation. Also, serum concentration of IL-6 is associated with airflow limitation.

Effect of Gender-Specific Adult Bovine Serum on Gene Expression During Myogenesis

  • Lee, Eun-Ju;Pokharel, Smritee;Kim, Jie-Hoe;Nam, Sang-Sup;Choi, In-Ho
    • Journal of Animal Science and Technology
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    • 제54권3호
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    • pp.219-226
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    • 2012
  • Gender specificity in muscle growth and development is well known. Genesis of muscle is dependent on proliferation and differentiation potential of resident myogenic satellite cells (MSCs) present in muscle fibers. Multipotential capacity of forming myocyte, osteocyte, and adipocyte like cell makes MSCs a unique stem cell. To understand the molecular mechanism involved in determination of muscle quality due to difference in hormone concentration of different gender of animals, MSCs were isolated from bovine skeletal muscle and cultured in male, female, and castrated serum supplemented media. DNA microarray used consisted of 24,000 spots with 70 mer oligo in each spot. A total of 88 genes were up-regulated and 551 genes were down-regulated by more than two fold. Among up-regulated gene, 33, 34, and 21 genes were found up-regulated in cells grown in male, female, and castrated serum, respectively. Interestingly, male serum showed 4, female 11 and castrated male showed 4 genes expressed highly in each gender. Further study on the highly up-regulated gene may unfold the mystery of gender specificity found in muscle development. Also, the identification of differentially expressed genes in gender-specific serum will add information on infrastructure of bovine genome research.

Rapid Detection of Serum HCV RNA by Combining Reverse Transcription and PCR without RNA Extraction

  • Jang, Jeong-Su;Lee, Kong-Joo
    • Archives of Pharmacal Research
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    • 제19권6호
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    • pp.486-489
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    • 1996
  • A simple, rapid, specific and sensitive method for the detection of serum hepatitis C virus (HCV) RNA using the reverse transcription-polymerase chain reaction (RT-PCR) technique without conventional RNA extraction was developed. HCV template RNA from serum was obtained by boiling the serum at $95^{\circ}C$ for 2 min, cooling rapidly in ice and removing the proteins by cetrifugation. RT-PCR amplifications including the reverse transcription and first PCR amplification were performed in one vessel containing both of reverse transcriptase and Taq DNA polymerase. The detection of HCV RNA from $10^{-3}{\mu}l$. serum was possible with this method. The suitability of this method for clinical analysis was evaluated by assaying HCV RNA in 225 patient samples including anti-HCV antibody negatives (13 samples) and positives (212 samples) by enzyme-linked immunosorbent assay test (ELISA). Detections of HCV RNA with this method were in 4 of 13 anti-HCV antibody negative samples (30.8%) and 95 of 212 positive samples (44.8%). The present method can be completed in 1 hr and has a wide range of application for the clinical utilities to determine the viral RNAS.

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청간해울탕(淸肝解鬱湯)이 생취에 Immobilization-Stress 및 Cold-Stress 부하후(負荷後) 혈중(血中)Histamine과 Corticosterone 함량(含量) 및 면역능(免疫能)에 미치는 영향(影響) (Effects of Chungganhaewooltang on Serum Levels of Histamine and Corticosterone and Immune Response after Immobilization-Stress or Cold-Stress in Mice)

  • 강복환;정우석;김송백;유심근
    • 대한한방부인과학회지
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    • 제18권4호
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    • pp.85-105
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    • 2005
  • Purpose : Investigate the effects of Chungganhaewooltang(CHT) on immobilization-stress or cold-stress in C576BL/6J mice. Methods : Male C57BL/6J 30 mice of weighting 18${\pm}$2g, were divided into sixs groups including the immobilization-stress group(5heads), after immobilization-stress CHT oral administration(500mg/kg) groups(5heads), cold-stress group(5heads) and after cold-stress CHT oral administration(500mg/kg) groups(5heads). then we observed changes in the serum histamine and corticosterone level and changes immune system Results : Immobilization-stress or cold-stress increased the serum level of histamine and corticosterone. CHT decreased the serum level of histamine and corticosterone increased by cold-stress. CHT inhibited the release of histamine from mast cells at the concentration of 0.1 mg/ml. In addition, immobilization-stress or cold-stress decreased the cell viability of murine thymocytes and splenocytes. CHT increased the cell viability of thymocytes decreased by immobilization-stress or cold-stress, but did not affect the cell viability of splenocytes decreased by immobilization-stress or cold-stress. Also immobilization-stress or cold-stress increased DNA fragmentation of thymocytes and splenocytes. CHT decreased DNA fragmentation of thymocytes increased by immobilization-stress or cold-stress, but did not affect DNA fragmentation of splenocytes increased by immobilization-stress or cold-stress. Immobilization-stress increased the population of thymic $CD4^+$ cells. CHT decreased the population of thymic $CD4^+$ cells increased by immobolization-stress. Immobilization-stress or cold-stress decreased the population of $B220^+$ cells and increased the population of $thy1^+$ cells. CHT decreased the population of $thy1^+$ cells increased by immobilization-stress or cold-stress. Immobilization-stress or cold-stress increased the population of splenic $CD4^+$ cells and $CD8^+$ cells. CHT decreased the population of splenic $CD4^+$ cells increased by immobolization-stress or cold-stress. Immobilization-stress or cold-stress decreased the production of ${\gamma}-interferon$(IFN) interleukin(IL)-2 and IL-4. CHT enhanced the production of ${\gamma}-IFN$ decreased by immobilization-stress or cold-stress but did not affect the production of IL-2 and IL-4 decreased by immobilization-stress or cold-stress. Furthermore, Immobilization- stress or cold-stress decreased the phagocytic activity of peritoneal macrophages and the production of nitric oxide. CHT enhanced the phagocytic activity and nitric oxide production decreased by cold-stress. Conclusion : CHT may be useful for the prevention and treatment of stress via suppression of serum histamine and corticosterone level and enhancement of immune response.

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출생전후의 식이 필수 지방산의 수준이 흰쥐의 혈청 및 뇌와 간세포의 Vitamin E 농도에 미치는 효과 (Effect of Pre-and Postnatal Feeding of Different Fats on Vitamin E Levels in Serum, Brain, and Liver of Rats)

  • 최민숙;송지현;최혜미;박현서;이양자
    • Journal of Nutrition and Health
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    • 제16권4호
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    • pp.287-295
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    • 1983
  • Sprague Dawley pregnant rats were fed the hydrogenated coconut oil (P/S 0.02), corn-oil plus beef tallow (P/S 1.0), and corn-oil(P/S 4.0)diet during the 14 days of gestation and through lactation. At weaning, offsprings from each group were divided into two and corn- oil diet was switched to coconut and vice versa, while the other corresponding groups continud the same diet. Body, brain, and liver weights were measured on pups and the brains were analyzed for DNA and vitamin E. Serum and liver vitamin E levels were alse measured. Body weight of coconut group was significantly low throughout the 7 week period, but brain weight was significantly depressed before weaning. Liver weight showed similar pattern with the body weight. Serum vitamin levels increased before weaning, while tissue vitamin E increased after weaning. Serum vitamin E levels of switched groups were similar to that of P/S 1.0 group. Brain vitamin E levels per DNA of switched groups were also similar to that of P/S 1.0 group. And the effect of different dietary essential fatty acid levels on vitamin E in the brain during lactation found to be insignificant.

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Cytotoxic Effect of Bee (A. mellifera) Venom on Cancer Cell Lines

  • Borojeni, Sima Khalilifard;Zolfagharian, Hossein;Babaie, Mahdi;Javadi, Iraj
    • 대한약침학회지
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    • 제23권4호
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    • pp.212-219
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    • 2020
  • Objectives: Nowadays cancer treatment is an important challenge in the medical world that needs better therapies. Many active secretions produced by insects such as honey bees used to discover new anticancer drugs. Bee venom (BV) has a potent anti inflammatory, anti cancer and tumor effects. The aim of present study is evaluation of anticancer effects induced by Apis mellifera venom (AmV) on cell Lines. Methods: AmV was selected for study on cancer cell lines. Total protein, molecular weight and LD50 of crude venom were determined. Then, cells were grown in Dulbecco's Modified Eagle medium supplemented with 10% fetal bovine serum and 1% antibiotics. The A549, HeLa and MDA-MB-231 cell Lines were exposed by different concentration of AmV. The morphology of cells was determined and cell viability was studed by MTT assay. Evaluation of cell death was determined by and DNA fragmentation. Results: The results from MTT assay showed that 3.125 ㎍/mL of A549, 12.5 for HeLa and 6.25 ㎍/mL of MDA-MB-231 killed 50% of cells (p < 0.05). Morphological analysis and the results from hoescht staining and DNA fragmentation indicated that cell death induced by AmV was significantly apoptosis. Conclusion: The data showed that using lower dosage of AmV during treatment period cause inhibition of proliferation in time and dose dependant manner. Findings indicated that some ingredients of AmV have anticancer effects and with further investigation it can be used in production of anticancer drugs.

소아 만성 B형 간염의 Interferon Alfa 치료 후 혈청학적, 조직학적 소견의 변화 (Serological and Histological Changes after Interferon Alfa Therapy in Children with Chronic Hepatitis B)

  • 고재성;정주영;장자준;서정기
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • 제3권1호
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    • pp.56-62
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    • 2000
  • 목 적: 만성 B형 간염 환자의 치료에 interferon alfa는 효과가 있는 것으로 알려지고 있지만 연구자에 따라 차이가 있으며 소아에서의 치료 후의 조직학적 변화에 대한 연구는 거의 없는 실정이다. 저자들은 소아 만성 B형 간염에서 interferon alfa의 치료 효과와 추적 생검이 가능했던 환자에서의 조직학적 변화에 대해 알아보고자 하였다. 대상 및 방법: 1995년 3월부터 1997년 8월까지 서울대학교병원 소아과에서 6개월 이상 HBsAg과 HBeAg 및 HBV DNA가 양성이었으며 간 조직검사상 만성 간염으로 확진된 환자 35명을 대상으로 recombinant interferon alfa 3~6 MU(평균 $3.4\;MU/m^2$)를 주 3회씩 6개월간 피하주사 하였다. 치료 시작 12개월 이상 혈청학적 변화를 추적 관찰하였다. interferon에 반응이 있는 환자 중 18명에서 치료후 간생검을 실시하여 조직학적 변화를 분석하였다. 결 과: 1) 치료 환자 35명 중 17명(49%)에서 인터페론 치료 시작 6개월에 HBV DNA의 감소가 있었고, 12개월째에 22명(63%)에서 HBeAg 및 HBV DNA의 음전이 생겼으며 18개월까지는 25명(71%)에서 관찰되었다. 반응군에서 혈청 ALT치는 모두 정상화되었고, HBsAg의 음전은 1명에서 관찰되었다. 2) 어머니가 HBsAg 보유자가 아닌 수평감염, 치료전 ALT가 정상의 2배 이상, 간조직의 심한 괴사와 염증이 interferon에 대한 반응이 좋은 예측인자이었다. 3) 치료 후 반응군의 간조직 소견에서 간맥주위괴사, 소엽내 활성도, 간맥내 염증, 간섬유화, total HAI가 유의하게 감소하였다. 결 론: 소아 만성 B형 간염에서 interferon alfa 치료 후 63%에서 반응을 보였으며, 혈청학적 변화는 조직학적 소견의 호전과 연관이 있다. 소아 만성B형 간염 환아에서 interferon 치료는 혈청학적, 생화학적, 조직학적 관해를 유도하는 효과적이고 안전한 치료방법이다.

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