• Title/Summary/Keyword: Serine protease inhibitor

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Analysis of Bovine Seminal Plasma Proteins from Korean Native Cattle, Hanwoo, and Korean Native Brindle Cattle

  • Lee, Su-Rok;Kim, Eun-Sung;Kim, Sung-Woo;Kim, Hyeong-Chul;Shim, Kwan-Seob;Kim, Jong-Gug
    • Reproductive and Developmental Biology
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    • v.36 no.2
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    • pp.121-131
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    • 2012
  • After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q-ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.

A Case of 47-Years-Old Female with Obstructive Jaundice and Weight Loss

  • Park, Pil Gyu;Kang, Huapyong;Chung, Moon Jae;Park, Jeong Youp;Bang, Seungmin;Park, Seung Woo;Song, Si Young;Lee, Hee Seung
    • Journal of Digestive Cancer Reports
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    • v.7 no.1
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    • pp.18-21
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    • 2019
  • Serine protease inhibitor Kazal-type 1 (SPINK1) is a gene expressed from pancreatic acinar cell which its mutation is known to be associated with chronic pancreatitis (CP) and pancreatic cancer. We report a case of a 47-years-old female with nausea and weight loss with yellow discoloration of skin. Initial imaging and endoscopic study led us to an impression of chronic pancreatitis with pancreatic cancer with common bile-duct dilation. Biopsy result was confirmed with pancreatic adenocarcinoma and additional imaging revealed lymph node and bone metastasis. Our genetic analysis revealed 194+2T>C mutation of SPINK1. Biliary obstruction was successfully decompressed by stent insertion and underwent chemotherapy and radiotherapy. Although there is accumulating evidence of association between SPINK1 mutation and CP, the relationship between SPINK1 mutation and pancreatic cancer in CP patient is an emerging concept. Genetic analysis should be considered in patients with young age especially when diagnosed with both CP and pancreatic cancer.

Comparison of Gene Expression Profile in Eutopic Endometria with or without Endometriosis: A Microarray Study (자궁내막증 환자와 대조군에서의 자궁내막 유전자 발현의 차이: Microarray를 이용한 연구)

  • Chung, Min-Ji;Chung, Eun-Jung;Lee, Shin-Je;Kim, Moon-Kyu;Chun, Sang-Sik;Lee, Taek-Hoo
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.1
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    • pp.19-31
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    • 2007
  • Objective: Pathogenesis of the endometriosis is very complex and the etiology is still unclear. Our hypothesis is that there may be some difference in gene expression patterns between eutopic endometriums with or without endometriosis. In this study, we analyzed the difference of gene expression profile with cDNA microarray. Methods: Endometrial tissues were gathered from patients with endometriosis or other benign gynecologic diseases. cDNA microarray technique was applied to screen the different gene expression profiles from early and late secretory phase endometria of those two groups. Each three mRNA samples isolated from early and late secretory phase of endometrial tissues of control were pooled and used as master controls and labeled with Cy3-dUTP. Then the differences of gene expression pattern were screened by comparing eutopic endometria with endometriosis, which were labeled with Cy5-dUTP. Fluorescent labeled probes were hybridized on a microarray of 4,800 human genes. Results: Twelve genes were consistently over-expressed in the endometrium of endometriosis such as ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ transporting (ATP5C1) and TNF alpha factor. Eleven genes were consistently down-regulated in the endometriosis samples. Many extracellular matrix protein genes (decorin, lumican, EGF-containing fibulin-like extracellular matrix protein 1, fibulin 5, and matrix Gla protein) and protease/protease inhibitors (serine proteinase inhibitor, matrix metalloproteinase 2, tissue inhibitor of metalloproteinase 1), and insulin like growth factor II associated protein were included. Expression patterns of selected eight genes from the cDNA microarray were confirmed by quantitative RT-PCR or real time RT-PCR. Conclusion: The result of this analysis supports the hypothesis that the endometrium from patients with endometriosis has distinct gene expression profile from control endometrium without endometriosis.

Maspin Expression and Its Clinical Significance in Non-Small Cell Lung Cancer (비소세포폐암에서 Maspin의 발현과 임상적 의의)

  • Yoon, Seong-Hoon;Kim, Won-Jin;Shin, Kyung-Hwa;Kim, Mi-Hyun;Cho, Woo-Hyun;Kim, Ki-Uk;Park, Hye-Kyung;Jeon, Doo-Soo;Kim, Yun-Seong;Lee, Chang-Hun;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.2
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    • pp.132-138
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    • 2011
  • Background: Maspin (mammary serine protease inhibitor) is a member of the serpin superfamily. A few studies have examined the role of maspin in tumor suppression of non-small cell lung cancer (NSCLC); however, its role in the development and progression of NSCLC still remains controversial. We evaluated the immunohistochemical expression of maspin in order to elucidate its clinical significance in NSCLC. Methods: We analyzed 145 patients with pathologically confirmed NSCLC, including 66 cases of squamous cell carcinomas (SCCs) and 79 cases of adenocarcinomas (ADCs). We performed a immuno-histochemical stain with maspin and PCNA (proliferating cell nuclear antigen) using tissue microarray blocks. Results: There were 108 men and 37 women in the study population. The mean age of patients in the study was 63.7 years (range, 40.0~82.0; median, 65.0). The proportion of maspin expression was significantly higher in SCCs (52/66, 78.8%; p<0.01) than in ADCs (17/79, 21.5%; p<0.01). Maspin expression was not associated with PCNA (p=0.828), lymph node involvement (p=0.483), or tumor stage (p=0.216), but showed correlation with well-to-moderate tumor differentiation (p=0.012). There was no observed correlation between maspin expression and survival with NSCLC (p=0.218). Conclusion: The present study suggests that maspin expression was significantly higher in SCCs than in ADCs and was associated with low histological grade. However, maspin expression was not an independent factor to predict a prognosis in NSCLC.

Changes of the Maternal Insulin-like Growth Factors System in Pregnant Rats During Perinatal Periods (주산기 랫드 모체에서 Insulin-like Growth Factor System의 변동)

  • Jin, Song-jun;Park, Soo-hyun;Cho, Nam-pyo;Kang, Chang-won
    • Korean Journal of Veterinary Research
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    • v.43 no.3
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    • pp.383-392
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    • 2003
  • Insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) are important regulators on the development of maternal tissues during pregnancy. This study was performed to examine the relationship between maternal IGFs/IGFBPs system (i.e: IGF-I, II, their receptors, and IGFBPs) in pre- and post-partum rats. The liver and kidney are important organs for the synthesis of IGFs and IGFBPs in adults. The levels of materanal IGFs and IGFBPs in serum, liver, and kidney were examined at 14 and 21 days of gestation and at 3, 7, 11, and 14 days after birth. The expression of IGFs and their receptors mRNA was also examined in fetal and maternal rat liver, kidney. IGF-I concentrations in maternal serum and liver were decreased during pregnancy. However, IGF-I concentration in maternal kidney was increased, having maximal effect at 14 days of gestation. IGF-I concentrations were decreased in serum, liver, and kidney of postpartum rat, compared to control (p < 0.05). On the other hand, IGF-II concentrations in serum, liver, and kidney were increased during pregnancy (p<0.05) and gradually decreased to control level in postpartum period. The levels of IGFBP-3 and IGFBP-2 are expressed in serum, liver, and kidney. However, IGFBP-3 is mainly expressed in serum and liver, and IGFBP-2 in kidney. The levels of IGFBP-3 and IGFBP-2 in maternal serum were markedly decreased during pregnancy and gradually recovered to control level during postpartum period by western ligand blotting. However, there was no change of IGFBP-3 and IGFBP-2 levels by western immunoblotting. The levels of IGFBP-3 and IGFBP-2 in maternal liver and kidney also showed the same pattern of serum, although the main IGFBP is different. In normal rat serum, IGF-I 150 kDa and 50 kDa carrier proteins were detected. The level of IGF-I 150 kDa carrier proteins in pregnant rat was decreased compared to normal rat, but that of 50 kDa carrier proteins was increased. IGFBP-3 protease activity was identified in pregnant rat serum and maternal placenta, and it was inhibited by EDTA ($Ca^{2+}$ chelating agent) and aprotinin (serine proteinase inhibitor). Taken together, these results suggest that the changes of IGFs and IGFBPs in maternal rats are regulated by liver and kidney IGFs and their receptors mRNA during the pregnancy.

Compound K improves skin barrier function by increasing SPINK5 expression

  • Park, No-June;Bong, Sim-Kyu;Lee, Sullim;Jung, Yujung;Jegal, Hyun;Kim, Jinchul;Kim, Si-Kwan;Kim, Yong Kee;Kim, Su-Nam
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.799-807
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    • 2020
  • Background: The skin acts as a barrier to protect organisms against harmful exogenous agents. Compound K (CK) is an active metabolite of ginsenoside Rb1, Rb2 and Rc, and researchers have focused on its skin protective efficacy. In this study, we hypothesized that increased expression of the serine protease inhibitor Kazal type-5 (SPINK5) may improve skin barrier function. Methods: We screened several ginsenosides to increase SPINK5 gene promoter activity using a transactivation assay and found that CK can increase SPINK5 expression. To investigate the protective effect of CK on the skin barrier, RT-PCR and Western blotting were performed to investigate the expression levels of SPINK5, kallikrein 5 (KLK5), KLK7 and PAR2 in UVB-irradiated HaCaT cells. Measurement of transepidermal water loss (TEWL) and histological changes associated with the skin barrier were performed in a UVB-irradiated mouse model and a 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis-like model. Results: CK treatment increased the expression of SPINK5 and decreased the expression of its downstream genes, such as KLKs and PAR2. In the UVB-irradiated mouse model and the DNCB-induced atopic dermatitis model, CK restored increased TEWL and decreased hydration and epidermal hyperplasia. In addition, CK normalized the reduced SPINK5 expression caused by UVB or DNCB, thereby restoring the expression of the proteins involved in desquamation to a level similar to normal. Conclusions: Our data showed that CK contributes to improving skin-barrier function in UVB-irradiated and DNCB-induced atopic dermatitis-like models through SPINK5. These results suggest that therapeutic attempts with CK might be useful in treating barrier-disrupted diseases.

Microarray Analysis of Gene Expression in RAW Cells Treated with Carthami Flos Herbal Acupuncture Solution (홍화약침액(紅花藥鍼液)이 RAW Cell 유전자발현(遺傳子發顯)에 미치는 영향(影響))

  • Kang, Seung-Beom;Kim, Jong-In;Kim, Yong-Seok;Kang, Sung-Keel;Koh, Hyung-Kyun
    • Journal of Acupuncture Research
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    • v.25 no.1
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    • pp.139-154
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    • 2008
  • 목적 : 홍화(紅花)는 활혈거어(活血祛瘀), 해독지통(解毒止痛)의 효능이 있어 관절염, 동맥경화(動脈硬化), 종양(腫瘍), 월경부조(月經不調), 뇌혈전(腦血栓)에 사용되어 왔다. 이에 홍화약침액(紅花藥鍼液)의 분자생물학적 효능 분석을 하고자 Lipopolysaccharide(LPS)로 염증을 유발한 RAW 264.7 cell의 유전자(遺傳子) 발현(發顯)에 미치는 영향을 Microarray를 통하여 관찰하였다. 방법 : RAW cell을 배양하고 홍화약침액(紅花藥鍼液)의 세포 독성을 확인한 후 (1) LPS, (2) 홍화약침액(紅花藥鍼液), (3) 홍화약침액(紅花藥鍼液)과 LPS를 처치했을 때의 유전자 발현양상을 microarray를 이용하여 관찰하였다. 대조군에 비해 2배 이상 발현의 차이가 있는 경우를 유의한 것으로 보았다. 결과 : 8,170개의 유전자 중 (1) LPS를 처치하였을 경우 35개의 유전자에서 발현이 상승되었고, (2) 홍화약침액(紅花藥鍼液)을 처치하였을 경우 11개의 유전자에서 발현이 상승되고 53개의 유전자에서 발현이 억제되었으며, (3) 홍화약침액(紅花藥鍼液)과 LPS를 동시에 처치하였을 경우에는 47개의 유전자에서 발현이 상승되었고 11개의 유전자에서 발현이 억제되었다. LPS 자극으로 발현이 상승되었지만 홍화약침액(紅花藥鍼液)을 처치할 때 발현이 억제되는 유전자는 SUMO1/sentrin specific protease 7(SENP7), Serine(or cysteine) proteinase inhibitor, clade B(ovalbumin), member 7(SERPINB7), M-phase phosphoprotein, mpp8(HSMPP8), Glycogenin 2(GYG2), InaD-like(Drosophila)(INADL), Copine III(CPNE3), Loss of heterozygosity, 11, chromosomal region 2, gene A(LOH11CR2A), Chromosome 9 open reading frame 33(SHC3), NADH dehydrogenase(ubiquinone) 1 beta subcomplex, 2, 8kDa(NDUFB2)로 9개가 있었다. 요약 : 홍화약침액(紅花藥鍼液)이 LPS로 염증을 유발시킨 RAW 264.7 cell의 유전자 발현에 미치는 영향을 Microarray를 통해 분석하였다. 홍화약침액(紅花藥鍼液)이 LPS로 발현을 항진시킨 35개의 유전자 중 9개를 효과적으로 억제하는 것을 확인하여 염증 치료 기전을 시사하는 유용한 자료를 얻을 수 있었으며 홍화약침액(紅花藥鍼液)이 발현을 항진시킨 유전자들을 통해 혈관생성과 종양억제 등 보다 넓은 범위에 대한 연구가 가능할 것으로 사료된다.

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Analysis of Differentially Expressed Genes by Resveratrol Using Membrane Microarray (Membrane Microarray를 이용한 Resveratrol에 의해 차별적으로 발현되는 유전자 군의 분석)

  • Kim, Jong-Sik;Jang, Min-Jung;Kim, Hyo-Eun;Kim, Soon-Young;Kim, Byung-Oh;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1115-1120
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    • 2007
  • In the present study, we investigated whether several phytochemicals (resveratrol, genistein, epicatechin gallate, dially disulfide, caffeic acid phenetyl ester) and sulindac sulfide could induce expression of tumor suppressor p53 protein in human colorectal HCT116 cells. We found that p53 was dramatically induced by all phytochemical treatments except sulindac sulfide. Among treated phytochemicals, we selected resveratrol for further experiments because it is one of the highest p53 inducer. Using a Western blot analysis, we found that resveratrol induced p53 in a dose- and time-dependent manner. Additionally, using membrane-based microarray analysis, we found that twenty-five genes were up-regulated and two genes were down-regulated by resveratrol treatment. Among the up-regulated genes, we selected 4 genes and performed reverse-transcription-PCR to confirm microarray data. The results of RT-PCR were highly accorded with those of membrane microarray. In addition, we found that thrombospondin-1 (TSP-1) expression was not dependent on p53 presence, whereas mammary serine protease inhibitor (MASPIN) expression was dependent on p53 expressed by resveratrol treatment. The results of this study may help to promote our understandings of the molecular mechanisms of chemoprevention that are mediated by resveratrol in human colorectal cancer.

The Use of Nafamostat Mesilate as an Anticoagulant during Continuous Renal Replacement Therapy for Children with a High Risk of Bleeding (출혈성 경향이 높은 소아환자의 지속성 신대체 요법시 사용되는 항응고제로서 Nafamostat mesilate의 사용)

  • Lee, Sang Taek;Cho, Heeyeon
    • Childhood Kidney Diseases
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    • v.18 no.2
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    • pp.98-105
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    • 2014
  • Purpose: Nafamostat mesilate (NM), a synthetic serine protease inhibitor, has been investigated as an anticoagulant for adult patients with a high risk of bleeding, who need chronic renal replacement therapy (CRRT). However, little is known about the use of NM as an anticoagulant in pediatric CRRT. The aim of this study was to evaluate the ideal dosage, efficacy, and safety of NM in pediatric CRRT. Methods: We conducted a retrospective study of 40 pediatric patients who had undergone at least 24 h of venovenous CRRTs between January 2011 and October 2013. We divided the patients according to risk of bleeding. Those at high risk received no anticoagulation (group 1) or NM as an anticoagulant (group 2), while those at low risk received heparin (group 3). Results: Forty patients (25 male and 15 female; mean age, $8.2{\pm}6.6$ years) were enrolled. The mean duration of CRRT was 13.0 days, and the survival rate was 57.5%. The mean hemofilter lifespan was 39.3 h in group 1 and 11.3 h in group 3. In group 2, hemofilter lifespan was extended from 7.5 h to 27.4 h after the use of NM (P =0.001). The mean hemofilter lifespan with NM was greater than with heparin (P =0.018). No patient experienced a major bleeding event during treatment with NM. Conclusion: NM may be a good alternative anticoagulant in pediatric patients with a high risk of bleeding requiring CRRT, and is not associated with bleeding complications.

Evaluation of Sleep Comfort for Indoor Thermal Environment based on the Physiological Signal Analysis (생리신호 분석을 이용한 실내온열환경에서의 수면 쾌적성 평가)

  • 이낙범;임재중;금종수;이구형;최호선
    • Science of Emotion and Sensibility
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    • v.3 no.2
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    • pp.75-84
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    • 2000
  • 문명의 발달과 함께 수면부족으로 인한 여러 가지 스트레스와 질환이 증가하게 되어 최근 수면연구에 대한 관심이 증가하고 있다. 본 연구는 다양한 온열환경 조건에서의 쾌적한 수면을 위한 온열환경을 제시하기 위해, 5명의 여성 피험자를 대상으로 22$^{\circ}C$, 26$^{\circ}C$, 3$0^{\circ}C$의 일정온도 조건과 $25^{\circ}C$에서 1시간 후와 2시간 후에 각각 1, 2$^{\circ}C$를 상승시켜주는 변동온도 조건하에서 수면생리신호를 측정하였다. 그리고, 수면단계 평가를 이용하여 총 수면시간, 깊은 수면의 비율, 그리고 최초 수면시작 시간에서 최초의 서파 수면이 나타나기까지의 지연시간 등의 수면효율을 평가하였다. 그 결과, 일정온도 조건에서는 26$^{\circ}C$에서 총 수면시간(466.7$\pm$10.25분)과 깊은 수면의 비율(33.1$\pm$4.95%)은 타 조건에 비해 높게 나타났고, 최초 서파수면까지의 지연시간(9.8$\pm$3.33분)은 타 조건에 비해 낮게 나타나 쾌적한 수면을 위한 가장 적절한 온열조건임을 관찰할 수 있었다. 그리고 변동온도 조건에서는 4가지 온열조건간에는 큰 차이가 나타나지 않았지만, 모든 조건에서 일정온도 조건보다는 좋은 결과를 나타내었다. 또한 수면 중 신체 움직임과 설문 분석에서도 동일한 결과를 보였다. 본 연구를 통해, 수면생리신호를 이용한 수면 쾌적성 분석은 수면의 질적인 상태를 관찰하는데 매우 적합한 파라메터를 도출할 수 있으며, 여러 가지 수면환경 조건을 평가하는데 매우 유용한 지표가 될 수 있음을 보였다.e results suggest that hCG treatment at 7 days after insemination could be used to increase the pregnancy rate of embryo transfer, and transfer, and only the recipients with PUN concentration of <12 mg/dl were influenced by treatment with hCG./TEX>이었으며, 이는 화성 기원을 지시한다.sucrose를 이용한 2단계 희석이 수정란의 생존성을 향상시키는 것으로 나타났다. 또한 발육 단계별 생존성에 있어서는 발육이 진전된 확장배 반포 시에 동결하는 것이 배반포기에 동결하는 것 보다 유리한 것으로 나타났다.ody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다..ments of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to

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