• Title/Summary/Keyword: NIH/3T3

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A Novel Heptapeptide that Promotes Cellular Activity and Inhibits Photoaging in Fibroblasts (섬유아세포에서 세포 활성 촉진 및 광노화 억제 효능을 보이는 신규 헵타펩타이드)

  • Lee, Eung Ji;Kang, Hana;Hwang, Bo Byeol;Chung, Yong Ji;Kim, Eun Mi
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.2
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    • pp.157-167
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    • 2022
  • In this study, we investigated the effects of heptapeptide on cellular activation and inhibition of cellular damage induced by photoaging condition in NIH3T3 fibroblasts. Cell proliferation and extracellular matrix (ECM) expression were induced by heptapeptide. The reduced cell viability under photoaging condition through ultraviolet A (UVA) irradiation was increased by heptapeptide. And UVA-induced apoptosis, matrix metalloproteinases-1 (MMP-1) expression, and reactive oxygen species (ROS) level were decreased by heptapeptide. In addition, the inhibition of transforming growth factor-β (TGF-β)/smad signaling under UVA irradiation which resulting in reduction of ECM expression was also recovered by heptapeptide. We also tested the effect of heptapeptide under another photoaging condition through heat shock, and pre-treatment of heptapeptide prevented the phosphorylation of mitogen-activated protein kinase (MAPK) and MMP-1 expression induced by heat shock. From these results, it has been shown that the heptapeptide has protective effects on fibroblasts through the up-regulation of cellular activity and through the decreasing of intracellular ROS level induced by UVA irradiation or heat shock. It is expected that the dermal protection effect of heptapeptide can be applied as a new cosmetic material in the future.

A case of Hyper-IgE syndrome with a mutation of the STAT3 gene (STAT3유전자 돌연변이 검사로 확진된 고면역글로불린E 증후군 1례)

  • Kang, Ji-Man;Suh, Jung-Min;Kim, Ji-Hyun;Kim, Hee-Jin;Kim, Yae-Jean;Lee, Hun-Seok;Shin, Young-Kee;Ahn, Kang-Mo;Lee, Sang-Il
    • Clinical and Experimental Pediatrics
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    • v.53 no.4
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    • pp.592-597
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    • 2010
  • Hyperimmunoglobulin E syndrome (HIES) is a rare immunodeficiency disease which is characterized by high serum IgE levels, eczema, and recurrent infections. Herein we present the case of a patient with HIES associated with STAT3 gene ($stat3$) mutation. A 16 year-old girl was admitted to our hospital due to hemoptysis caused by pneumonia with bronchiectasis. She had a history of recurrent skin and respiratory tract infections, such as pneumonia caused by MRSA (methicillin-resistant $Staphylococcus$ $aureus$) and $Pseudomonas$ $aeruginosa$. On physical examination, a broad round shaped nose, oral thrush, and chronic eczematous skin rash over her whole body were found. Laboratory data showed an elevated eosinophil count ($750/{\mu}L$) and total IgE level (5,001 U/mL). The patient's National Institutes of Health (NIH) score for HIES was 44. Direct sequencing of the STAT3 gene revealed that the patient was heterozygous for a missense mutation in the DNA binding domain of the STAT3 protein (c.1144C>T, p. Arg382Trp). HIES should be suspected in patients with recurrent infections and can be confirmed by clinical scoring and genetic analysis.

Optimization of Retrovirus Mediated-Gene Transfer into Hematopoietic Stem Cells (Retrovirus를 이용하여 조혈모세포에 유전자를 전달하기 위한 최적화)

  • Kim, Sang-Gyeong;Seo, Heon-Seok;Lee, Jong-Won;Sin, Dong-Geon;Lee, Jae-Gwan;Kim, Hyeon-Min;Kim, Jae-Sik;Seo, Jang-Su
    • KSBB Journal
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    • v.14 no.5
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    • pp.593-599
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    • 1999
  • In this study, optimal conditions to infect CD34 positive cells containing hematopoietic stem cells obtained from cord blood and bone marrow were found using two different retroviral vectors expressing human growth hormone (hGH) and $\beta$-galactosidase. CD34 positive cells were successfully infected with recombinant retroviruses only when the CD34 positive cells were co-cultured with packaging cells secreting recombinant retroviruses. To find the highest infection efficiency for the gene transfer, CD34 positive cells from cord blood were co-cultured with packaging cells secreting recombinant retroviruses encoding E. coli lacZ gene. The highest infection efficiency was obtained when CD34 positive cells were cultured for 3 days, and then co-culturing was done for another 2 days. When CD34 positive cells from bone marrow were co-cultured with packaging cells secreting recombinant retroviruses encoding hGH gene, the maximum amount of hGH was also secreted at the same conditions found above, i.e. 3 days of culture and 2 days of co-culture. These results show that there are optimal conditions for the gene transfer into hematopoietic stem cells regardless of sources of target cells or retroviral vectors used to infect.

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Biological Activities of Calcium Polyphosphate (Calcium polyphosphate의 생물학적 활성도에 관한 연구)

  • Seol, Yang-Jo;Lee, Jae-Il;Lee, Yong-Moo;Lim, Yoon-Tak;Kim, Seok-Young;Ku, Young;Rhyu, In-Chul;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.213-231
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    • 2000
  • 이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

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Study on Performance Evaluation of Dental X-ray Equipment (치과 방사선 발생기의 성능평가에 관한 연구)

  • Jung, Jae-Eun;Jung, Jae-Ho;Kang, Hee-Doo;Lee, Jong-Woong;Ra, Keuk-Hwan
    • Korean Journal of Digital Imaging in Medicine
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    • v.11 no.2
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    • pp.115-119
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    • 2009
  • I think this will be valuable reference for assuring consistency and homogeneity of clarity and managing dental radiation equipment by experimentation of dental radiation equipment permanent which based on KS C IEC 61223-3-4 standard and KS C IEC 61223-2-7. Put a dental radiation generator and experiment equipment as source and film(sensor) length within 30 em, place the step-wedge above the film(sensor). Tie up tube voltage 60 kVp, tube current 7 mA and then get an each image through CCD sensor and film by changing the exposure time as 0.12sec, 0.25sec, 0.4sec. Repeat the test 5times as a same method. Measure the concentration of each stage of film image, which gained by experiment, using photometer. And the image that gained by CCD sensor, analyze the pixel value's change by using image J, which is analyzing image program provided by NIH(National Institutes of Health). In case of film, while 0.12sec and 0.25sec show regular rising pattern of density gap as exposure time's increase, 0.4sec shows low rather than 0.12sec and 0.25sec. In case of CCD sensor density test, the result shows opposite pattern of film. This makes me think that pixels of CCD's sensor can have 0~255 value but it becomes saturation if the value is over 255. The way that getting clear reception during decreasing human's exposed radiation is one of maintaining an equipment as a best condition. So we should keeping a dental radiation equipment's condition steadily through cyclic permanent test after factor examination. Even digital equipment doesn't maintain a permanent, it can maintain a clarity by post processing of image so that hard to set it as standard of permanent test. Therefore it would be more increase the accuracy that compare a film as standard image. Thus I consider it will be an important measurement to care for dental radiation equipment and warrant homogeneity, consistency of dental image's clarity through comparing pattern which is the result from factor test against cyclic permanent test.

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Melanogenesis Inhibitory and Antioxidant Activities of Phellinus baumii Methanol Extract (장수진흙버섯 메탄올 추출물의 멜라닌 생성 저해작용)

  • Lee, J.S.;Shin, D.B.;Lee, S.M.;Kim, S.H.;Lee, T.S.;Jung, D.C.
    • The Korean Journal of Mycology
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    • v.41 no.2
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    • pp.104-111
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    • 2013
  • Phellinus baumii is a medicinal mushroom used in Asian countries for a long period of time. The purpose of this study was to investigate the skin whitening activities of methanol extracts from fruiting bodies of P. baumii. To evaluate the antioxidant activities of the extract, polyphenol and flavonoid contents, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and chelating activity on ferrous ions were studied. For assay of skin whitening activities, tyrosinase and DOPA inhibitory activities, and tyrosinase and melanin synthesis inhibitory activities of B16/F10 melanoma cells treated with the methanol extract were investigated. The total polyphenol content of P. baumii methanol extract was 4.19. DPPH scavenging ability of P. baumii methanol extract was 88.26% in $25{\mu}g/mL$ concentration. We tested tyrosinase inhibitory activity and melanin contents in melanoma cells. The tyrosinase activity was inhibited to 65.17% at the concentration of $125{\mu}g/mL$ and melanin synthesis was inhibited to 57.61% at the concentration of $25{\mu}g/mL$. Overall, the experimental results showed that P. baumii methanol extract had inhibitory activities of tyrosinase and melanin synthesis by dose dependent manner in B16/F10 melanoma cells. Strong ultra-violet absorption spectra in the range of 270~370 nm indicated that ethanol extract of P. baumii could protect the skin from UV. Therefore, P. baumii methanol extract might be used for development of skin whitening, anti-UV and skin care agents.

EFFECT OF NERVE GROWTH FACTOR GENE INJECTION ON THE NERVE REGENERATION IN RAT LINGUAL NERVE CRUSH-INJURY MODEL (백서 설신경 압박손상모델에서 신경성장인자 유전자 주입이 신경재생에 미치는 영향)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Soung-Min;Kim, Yun-Hee;Jahng, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.5
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    • pp.375-395
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    • 2006
  • Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

Radio-isotope Red Cell Survival Studies Recommended by ICSH (방사성동위원소(放射性同位元素)를 이용(利用)한 적혈구수명측정법(赤血球壽命測定法)(ICSH추천(推薦)))

  • The Korea Society of Nuclear Medicine The Korea Society of Nuclear Medicine
    • The Korean Journal of Nuclear Medicine
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    • v.5 no.1
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    • pp.1-10
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    • 1971
  • 혈액질환(血液疾患), 특히 용혈성빈혈(溶血性貧血)을 수반(隨伴)한 경우(境遇)에 적혈구(赤血球)의 생성(生成) 및 파괴과정(破壞過程)을 정확(正確)히 파악(把握)하는 것은 중요(重要)하여 특히 적혈구수명측정(赤血球壽命測定)은 빈혈(貧血)의 본능(本能) 및 발생기전(發生機轉)을 이해(理解)하는데는 물론 병인적(病因的) 치료(治療) 및 예후(豫後)를 결정(決定)하는데 대단(犬端)히 유용(有用)하다. 적혈구수명측정(赤血球壽命測定)에는 1919년(年) Ashby가 개발(開發)한 differential agglutination법(法)이 이용(利用)되어 왔으나 수혈(輸血)에 따른 위험(危險)이 있고 방법(方法)이 복잡(複雜)하다는 단점(短點)을 가져 새로운 적혈구수명측정법(赤血球壽命測定法)이 연구(硏究)되어 왔다. 최근(最近)에 $^{51}Cr$이나, $^{32}DFP$같은 방사성동위원소(放射性同位元素)를 이용(利用)한 방법(方法)이 도입(導入)된 이래(以來) 임상적(臨床的)으로 적혈구수명측정(赤血球壽命測定)이 많이 시행(施行)되고 있지만 아직까지도 그 방법(方法)이나 결과(結果)의 해석(解釋)에 표준화(標準化)가 안되어 있다. 현재(現在) 임상영역(臨床領域)에서 가장 널리 이용(利用)되고 있는 적혈구수명측정법(赤血球壽命測定法)은 방사성(放射性) chromium($^{51}Cr$)법(法)으로 1950년(年) Gray와 Sterling에 의(依)해 창안(創案)된 이래(以來) 많은 학자(學者)들에 의(依)해 여러 가지 변법(變法)이 고안(考案)되어 왔는데, 이의 가장 큰 이유(理由)는 $^{51}Cr$이 적혈구표지(赤血球標識)에 가장 이상적(理想的)인 것만은 아니고 그 결과(結果)에 많은 요인(要因)들이 영향(影響)을 미치기 때문이다. 또 이런 변법(變法)의 사용(使用)은 각(各) 검사(檢査)에서 계산(計算)된 측정치(測定値)에 차이(差異)가 있어 그 결과(結果)의 해석(解釋) 및 비교(比較) 검토(檢討)에 적지않은 난점(難點)이 생겨 표준화(標準化)된 공통적(共通的)인 방법(方法)의 사용(使用)이 중요(重要)하다는 사실(事實)이 인식(認識)되게 되었다. 1966년(年) 호주(濠洲)의 Sydney에서 개최(開催)되었든 제11차(第11次) 국제혈액학회(國際血學會)때 열린 제4차(第4次) International Committee for Standardization in Haematology(ICSH)에서 Diagnostic Applications of Radioisotopes in Haematology에 관(關)한 expert panel을 갖을것을 의결(議決)하여 다음과 같은 12명(名)의 위원(委員)이 결정(決定)되었으며 위원회(委員會)의 의장(議長)에 Dr. Szur, 총무(總務)에 Dr. Glass가 각각(各各) 선임(選任)되었다. 그간(間) 1967년(年) 영경(英京) London에서 첫 회합(會合)이 있은후(後) New York, Vienna(IAEA후원(後援)) Brthesda(NIH후원(後援))에서 전문위원회(專門委員會)를 갖고 적혈구수명측정법(赤血球壽命測定法)에 관(關)한 의견(意見)의 일치(一致)를 보았다. ICSH와 국제혈액학회(國際血學會)에서는 이번에 결정(決定)된 적혈구수명측정법(赤血球壽命測定法)을 널리 소개(紹介)하며, 측정법(測定法)과 얻어진 결과(結果)의 해석(解釋)에 표준화(標準化)를 기(期)할 목적(目的)으로 이에 연관성(聯關性)있는 전문지(專門誌)에 게재(揭載)할 것을 요청(要請) 받었기에 이에 전문(全文)을 소개(紹介)하는 바이다. 이들은 방사성(放射性) chromium 법(法)의 모든 세부적(細部的)인 면(面)을 표준화(標準化)하고 있으며 그간(間) 가장 논란(論難)의 대상(對象)이 되었던, $^{51}Cr$-표지방법(標識方法)에 있어서의 세가지 변법(變法), 즉 ACD법(法), Citrate-wash법(法), ACD/ascorbic acid법(法)을 모두 인정(認定)하고 있다. 또한, DFP($DF^{32}P$ 또는 $^3H-DFP$) 표지법(標識法)의 표준방법(標準方法)도 기술(記述)하고 있으며, 더욱 중요(重要)한 것은 적혈구수명측정법(赤血球壽命測定法)으로서 현재(現在)까지 대부분(大部分)의 학자(學者)가 사용(使用)하여 왔던 지표(指標)인 $T_{50}Cr$ 대신(代身)에 mean red cell life span을 사용(使用)할 것을 권(勸)하고 있다.

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