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Possibility of Repeated Use of Elite Donor Cows for Mass Production of OPU-Derived Embryos (OPU 유래 수정란의 대량생산을 위한 고능력 공란우 반복사용 가능성에 관한 연구)

  • Jin, Jong-In;Choi, Byung-Hyun;Kim, Seong-Su;Park, Bun-Young;Lee, Jung-Gyu;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.149-159
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    • 2015
  • This study was designed to know the possibility in repeat uses of elite donor cows for getting mass production of OPU-derived embryo production (OPU-IVP). Ultrasound transvaginal ovum pick-up (OPU) performed in 6 Korean native cows was aged 4 to 10 years old. The aspiration of immature oocytes for OPU derived embryo was carried out 2 times per week, and OPU-IVP of $1^{st}$ period was carried out 22~48 sessions from each donors. And the break time for OPU-IVP of $2^{nd}$ period after $1^{st}$ OPU from each donors were 2~25 months. The OPU-IVP of $2^{nd}$ period each donors conducted total 15~65 times for 2~8 months by an ultrasonographic, was guided follicular aspiration system. The average numbers of collected oocytes, grade 1 + grade 2(G1+G2) oocytes and cleavage embryo from $1^{st}$ period OPU-IVP were significantly differences between donors (p<0.05). Total collected oocytes of donor D were significantly higher compared with donors of A, B, C, E and F (average 17.0 per session vs. 11.2, 10.1, 8.5, 10.2 and 9.6; p<0.05) and also oocytes of G1+G2 were significantly higher compared with r A and D and subsequently to donors of B, C, E and F (average 7.9 and 8.5 per session vs. 5.0, 2.7, 6.0 and 1.6; p<0.05). Cleavage rate of donor D was significantly higher compared with donors of A, B, C, E and F (average 13.1 per session vs. 10.1, 9.1, 6.9, 8.9 and 6.7; p<0.05). The average numbers of OPU-IVP for $1^{st}$ period was significantly higher from donors of B, D and E than those from donors of A, C and F (average 6.5, 7.1 and 6.5 per session vs. 3.5, 4.2 and 2.8; p<0.05). The possibility investigation of $2^{nd}$ OPU-IVP was carried out after 2~25 months rest periods from $1^{st}$ period OPU session. Total average numbers of collected oocytes, cleavages and blastocyst development rates were significantly higher from $1^{st}$ period OPU compared with $2^{nd}$ period one (p<0.05). The OPU-IVP efficiency by break for more embryo production from elite cow was analysis comparing without rest of donor A, under 6 months rest period as B and over 6 months rest period as C and then the average numbers of collected oocytes, cleavages and blastocysts were significantly higher from A group (11.8, 9.5 and 5.2 per session) than those from B and C groups (7.9, 6.2 and 2.6 vs. 9.2, 7.5 and 3.9, p<0.05), and also C group was significantly higher than B group. In conclusion, $1^{st}$ period OPU-IVP was more efficient compared with $2^{nd}$ period repeated uses of donor, and the break times for additional production of embryo on donor were needed more than over 6 months after $1^{st}$ period OPU-IVP. This repeating uses of elite donor cows given more emphasis for getting the opportunity on mass production of elite cow OPU-IVP embryo should be increased G1+G2 possibility of genetic improvement of livestock within short period.

Preparation and Evaluation of a 4-Branched Polyethylene Glycol Derivative Modified with Exendin-4 and Stearylamine for Extended Hypoglycemic Action

  • Kim, In-Soo;Ma, Kyung-Wan;Bae, Sung-Ho;Yoon, Jeong-Hyun;Oh, Kyung-Taek;Lee, Eun-Seong;Lee, Don-Haeng;Lee, Kang-Choon;Youn, Yu-Seok
    • Journal of Pharmaceutical Investigation
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    • v.40 no.3
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    • pp.175-180
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    • 2010
  • Albumin-modification has been viewed as one of the most effective ways of extending the short in vivo lifetimes of peptide drugs by delaying glomerular filtration. In this study, we describe a new type 2 anti-diabetic exendin-4 (Ex4) peptide derivative with significant binding ability to human serum albumin (HSA). This exendin-4 derivative consists of a 4-branched polyethylene glycol $(PEG)_{5k}$ (Mw: 20 kDa) modified with three stearylamines ($C_{18}-NH_2$) and one exendin-4 on its branches. PEG and stearylamine were selected to provide functionality to increase molecular size and bind to albumin, respectively. This derivative ($3C_{18}-4PEG_{5k}$-Ex4) was shown to have larger molecular size (Ca. 152 kDa) than actual (25.0 kDa) when subjected to size-exclusion chromatography, and the fluorescein-tagged $3C_{18}-4PEG_{5k}$-Ex4 displayed significant binding to the HSA-immobilized Sepharose CL-4B resin using confocal laser scanning microscopy. Furthermore, $3C_{18}-4PEG_{5k}$-Ex4 was found to have acceptable anti-hyperglycemic efficacy via three consecutive oral glucose tolerance testings (OGTT) in fasted type 2 diabetic db/db mice. The $HD_{total}$ value ($57.6{\pm}12.3%$) of $3C_{18}-4PEG_{5k}$-Ex4 at a 50 nmol/kg dose was 2-fold greater than that ($31.0{\pm}8.7%$) of native exendin-4 in non-fasted db/db mice. Especially, the blood glucose levels in the mice group treated with $3C_{18}-4PEG_{5k}$-Ex4 did not rebound to ~150 mg/dL until 24 h after the injection, which obviously shows the extended hypoglycemia. We believe that this derivative has great pharmaceutical potential as a novel long-acting type 2 anti-diabetic injection treatment.

Cytogenetic Analyses of Astragalus Species (황기류 식물 3종의 세포유전학적 분석)

  • Kim, Soo-Young;Choi, Hae-Woon;Kim, Chan-Soo;Sung, Jung-Sook;Lee, Joong-Ku;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.4
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    • pp.250-254
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    • 2006
  • To elucidate cytogenetic differences, karyotype analysis and FISH (fluorescence in situ hybridization) with 45S and 5S rDNAs were carried out in the three Astragalas species: Astragalas membranaceus Bunge, A. membranaceus var. alpinus Nakai and A. mongholicus Bunge. The somatic metaphase chromosome numbers of all three species were 2n=2x=16 and the size of chromosomes ranged $2.19{\sim} 5.73\;{\mu}m$. The chromosome complement of A. membranaceus consisted of each four pairs of metacentrics (chromosomes 3,4,6 and 7) and submetacentrics (chromosomes 1,2,4 and 8). In A. membranaceus var. alpinus, the chromosome complement consisted of two pairs of metacentrics (chromosomes 4 and 8) and six pairs of submetacentrics (chromosomes 1,2,3,5,6 and 7). A. mongholicus had three pairs of metacentrics (chromosomes 6,7 and 8) and five pairs of submetacentrics (chromosomes 1,2,3,4 and 5). Using bicolor-FISH, one pair of 45S and 5S rDNA signals could be detected on the centromeric regions of chromosomes 8 and 7 of A. membranaceus and A. mongholicus, respectively. In contrast, A, membranaceus var. alpinus had one pair of 45S signals on the centromeric region of chromosome 8 and two pairs of 5S rDNA signals on the short arms of chromosomes 7 and 8.

Effect of Heat-Treat Methods on the Soluble Calcium Levels in the Commercial Milk Products

  • Yoo, Sung-Ho;Kang, Seung-Bum;Park, Jin-Ho;Lee, Kyung-Sang;Kim, Jin-Man;Yoon, Sung-Sik
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.369-376
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    • 2013
  • Milk is well known to be rich in some nutrients such as protein, calcium, phosphorus, and vitamins. In particular, absorption and bioavailability of calcium receive lots of attention because calcium is very little absorbed until it is changed to the ionized form in the intestine. In this study, concentration of the soluble calcium was determined in the commercial bovine milk products, which were processed by different heat-treatment methods for pasteurization. As for general constituents, lactose, fat, protein, and mineral were almost same in the liquid milk products by different processors. Ultrafiltration of the skimmed milk caused little change in the permeate as for lactose content but both fat and protein decreased. pH values ranges from 6.57-6.62 at room temperature and slightly increase after centrifugation, 10,000 g, 10 min. Rennet-coagulation activity was the lowest in the ultra high temperature (UHT-)milk compared to the low temperature long time (LTLT-) and high temperature short time (HTST-)milk products. Each bovine milk products contains 1056.5-1111.3 mg/kg of Ca. The content of sulfhydryl group was the lowest in raw milk compared to the commercial products tested. For the skimmed milks after ultrafiltration with a membrane (Mw cut-off, 3 Kd), soluble Ca in the raw milk was highest at 450.2 mg/kg, followed by LTLT-milk 336.4-345.1 mg/kg, HTST-milk 305.5-313.3 mg/kg, UHT-milk 370.3-380.2 mg/kg in the decreasing order. After secondary ultrafiltration with a membrane (Mw cut-off, 1 kD), total calcium in raw milk had a highest of 444.2 mg/kg, and those in the market milk products. As follow: UHT-milk, 371.3 to 378.2 mg/kg; LTLT-milk, 333.3 to 342.2 mg/kg; HTST-milk 301.9 to 311.2 mg/kg in a decreasing order.

Effect of Soil Salinity on Nitrogen Mineralization of Livestock Manure Compost in Salt-Affected Coastal Soils

  • Kim, Jung-Hyun;Shim, Myung-Yong;Moon, Tae-Il;Kim, Seung-Hwan;Shin, Kook-Sik;Sonn, Yeon-Kyu;Chung, Doug-Young;Lee, Sang-Eun
    • Korean Journal of Soil Science and Fertilizer
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    • v.47 no.3
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    • pp.199-204
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    • 2014
  • We conducted a short-term incubation experiment in order to understand the effect of the salinity of reclaimed coastal soils on nitrogen mineralization of livestock manure compost (LMC). Two soils with the same soil texture but different EC levels were collected from the same field. These samples were treated with 0%, 1%, 2%, and 3% of LMC by weight basis and incubated at $25^{\circ}C$ to observe changes in inorganic N contents, pH, and dehydrogenase activity with respect to time. As a result, regardless of the soil EC level, as the LMC increased, the total content of the inorganic N ($NH_4{^+}+NO_3{^-}$) increased. Difference in the soil EC level did not affect N mineralization of LMC greatly. The soil EC had negligible effect on the dehydrgenase activity as with the case of inorganic nitrogen. The $NH_4{^+}$ contents remained very low throughout the experimental period starting from the first week of incubation. We believe this is due to the high pH level (pH 7.9 and pH 8.3) of the original soils leading to ammonia volatilization. On the other hand the $NO_3{^-}$ content maintained high level as the LMC treatment level increased and reached maximum at the third week. The pH of the soil during incubation period decreased as the $NO_3{^-}$ contents increased and increased slightly after three weeks. The rise of pH level is believed to be from the $NO_3{^-}$ absorption for immobilization by microbes. In conclusion, the high soil $EC_{1:5}$ level of $12dS\;m^{-1}$ conducted in this experiment did not affect the growth in terms of soil microbes involved in N mineralization of LMC.

Genetic Studies in Some Agronomic Characters in Rapeseed (Brassica napus L.) 1. Genetic analysis for maturity, plant height and grain weight per plant. (유채의 실용형질에 대한 유전연구 -제 1 보 유채의 초장, 성숙기 및 종실중의 유전분석-)

  • Lee, J.I.;Kwon, B.S.;Chae, Y.A.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.1
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    • pp.84-88
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    • 1984
  • To obtain basic information on the breeding of early maturing, short plant height rapeseed varieties, the following 7 varieties, Isuzu, Miyuki, Norin 25, Rang, Yongdang, Cresus and Tower were used in diallel crosses in 1979. Maturing date, plant height and grain weight per plant for the parents, $F_1$'s and $F_2$'s of the 7 x 7 partial diallel crosses were measured in 1981 for analysis of their genetic behavior. The results obtained are summarized as follows: 1. The days to maturing of $F_1$'s showed complete dominance for early maturing, and both additive and dominance genetic variances were significant. Number of effective factors in $F_1$'s was 3, but in $F_2$'s was 1. The degree of dominance in $F_1$'s was partial, while in $F_2$'s was complete. Both broad and narrow sense herita-bilities in $F_1$'s was high, while in $F_2$'s was low. 2. Yield per plant in $F_2$'s was controlled by additive component of genetic variance only, but $F_1$'s was different. The degree of dominance in $F_1$'s was complete, while in $F_2$'s was partial. The direction of dominance showed almost complete dominance over high yield and three effective factors was estimated. Yield per plant was controlled by recessive genes. 3. The plant height was controlled by both dominance and additive variance. Dominance was directed toward tall plant height. Number of effective factors was 2, and broad and narrow sense heritability were high in the plant height.

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Effect of Plant Growth Regulator Treatments on the Growth and Lateral Root Formation in Soybean Sprouts - I. Effect of Plant Growth Regulator Treatments on the Growth in Soybean Sprouts (생장조절물질(生長調節物質) 처리(處理)가 콩나물의 생육(生育) 및 세근발생(細根發生)에 미치는 영향(影響) - I. 생장조절물질(生長調節物質)의 단용(單用) 및 혼용처리(混用處理)가 콩나물의 생육(生育)에 미치는 효과(效果))

  • Kang, C.K.;Lee, J.M.;Saka, H.
    • Korean Journal of Weed Science
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    • v.9 no.1
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    • pp.56-68
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    • 1989
  • aA series of experiments were conducted to investigate the effect of plant growth regulator treatments on the growth and lateral root formation in soybean sprouts in order to establish the effective method of producing root-less or short-rooted soybean sprouts with larger diameter in the hypocotyl. Major results can be summarized as follows. 1. Soybean sprouts showed fairly uniform elongation rate from 3 to g days after imbibition with daily increase of 3.8cm. The speed of elongation of hypocotyl was reduced whereas that of root accelerated 7 days after imbibition. Lateral roots began to emerge fairly evenly from 5 to 9 days after imbibition with a daily increase of 4.4. 2. Auxins(IAA, IBA, NAA, 2,4-D) inhibited hypocotyl elongation and formation of lateral roots and increased hypocotyl diameter without influencing root length and hook diameter at higher concentrations. The dry weight of cotyledon was increased significantly as compared to that of hypocotyl and root. Among the tested auxins, 2, 4-D was the most effective. 3. BA and 4PU-30 significantly reduced elongation of hypocotyl and root and resulted in the biggest diameter of hypocotyl when treated at higher concentrations. The lowest effective concentration of BA to prevent the formation of larval gal roots was 12.5ppm. The formation of lateral roots could be completely prevented by BA and 4PU-30 treatment but kinetin, zeatin, zeatin riboside resulted in many lateral roots and increased thickness of soybean sprouts with little influence. Cotyledon deformation was found in soybean sprouts treated by 4PU-30. 4. 2, 4-D was the most effective for increasing the hypocotyl diameter while 4PU-30 was the most effective for reducing no. of lateral roots. 5. It can be concluded that among the plant growth regulators tested, BA was effective in reducing root length and increasing hypocotyl diameter. BA 12.5 ppm or 15 ppm may thus be the more practical for production of soybean sprouts. 6. ABA showed no significant effect of growth parameter, however ABA 25 ppm inhibited only no of lateral roots with little influence on the growth of seedling. 7. Ethephon inhibited the elongation of hypocotyl and root and increased hypocotyl diameter at higher concentrations. 8. The combined effect of cytokinins and ethephon was very similar to result of BA treatment alone. As the ethephon concentration increased, hypocotyl diameter and dry weight of cotyledon tended to increase.

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Radiation Dose-escalation Trial for Glioblastomas with 3D-conformal Radiotherapy (3차원 입체조형치료에 의한 아교모세포종의 방사선 선량증가 연구)

  • Cho, Jae-Ho;Lee, Chang-Geol;Kim, Kyoung-Ju;Bak, Jin-Ho;Lee, Se-Byeoung;Cho, Sam-Ju;Shim, Su-Jung;Yoon, Dok-Hyun;Chang, Jong-Hee;Kim, Tae-Gon;Kim, Dong-Suk;Suh, Chang-Ok
    • Radiation Oncology Journal
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    • v.22 no.4
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    • pp.237-246
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    • 2004
  • Purpose: To investigate the effects of radiation dose-escalation on the treatment outcome, complications and the other prognostic variables for glioblastoma patients treated with 3D-conformal radiotherapy (3D-CRT). Materials and Methods: Between Jan 1997 and July 2002, a total of 75 patients with histologically proven diagnosis of glioblastoma were analyzed. The patients who had a Karnofsky Performance Score (KPS) of 60 or higher, and received at least 50 Gy of radiation to the tumor bed were eligible. All the patients were divided into two arms; Arm 1, the high-dose group was enrolled prospectively, and Arm 2, the low-dose group served as a retrospective control. Arm 1 patients received $63\~70$ Gy (Median 66 Gy, fraction size $1.8\~2$ Gy) with 3D-conformal radiotherapy, and Arm 2 received 59.4 Gy or less (Median 59.4 Gy, fraction size 1.8 Gy) with 2D-conventional radiotherapy. The Gross Tumor Volume (GTV) was defined by the surgical margin and the residual gross tumor on a contrast enhanced MRI. Surrounding edema was not included in the Clinical Target Volume (CTV) in Arm 1, so as to reduce the risk of late radiation associated complications; whereas as in Arm 2 it was included. The overall survival and progression free survival times were calculated from the date of surgery using the Kaplan-Meier method. The time to progression was measured with serial neurologic examinations and MRI or CT scans after RT completion. Acute and late toxicities were evaluated using the Radiation Therapy Oncology Group neurotoxicity scores. Results: During the relatively short follow up period of 14 months, the median overall survival and progression free survival times were $15{\pm}1.65$ and $11{\pm}0.95$ months, respectively. The was a significantly longer survival time for the Arm 1 patients compared to those in Arm 2 (p=0.028). For Arm 1 patients, the median survival and progression free survival times were $21{\pm}5.03$ and $12{\pm}1.59$ months, respectively, while for Arm 2 patients they were $14{\pm}0.94$ and $10{\pm}1.63$ months, respectively. Especially in terms of the 2-year survival rate, the high-dose group showed a much better survival time than the low-dose group; $44.7\%$ versus $19.2\%$. Upon univariate analyses, age, performance status, location of tumor, extent of surgery, tumor volume and radiation dose group were significant factors for survival. Multivariate analyses confirmed that the impact of radiation dose on survival was independent of age, performance status, extent of surgery and target volume. During the follow-up period, complications related directly with radiation, such as radionecrosis, has not been identified. Conclusion: Using 3D-conformal radiotherapy, which is able to reduce the radiation dose to normal tissues compared to 2D-conventional treatment, up to 70 Gy of radiation could be delivered to the GTV without significant toxicity. As an approach to intensify local treatment, the radiation dose escalation through 3D-CRT can be expected to increase the overall and progression free survival times for patients with glioblastomas.

Composition Ratio Analysis of Transesterification Products of Olive Oil by Using Thin Layer Chromatography and Their Applicability to Cosmetics (올리브 오일의 에스터 교환반응 생성물의 TLC를 이용한 조성비 분석 및 화장품에의 응용가능성 평가)

  • Park, So Hyun;Shin, Hyuk Soo;Kim, A Rang;Jeong, Hyo Jin;Xuan, Song Hua;Hong, In Kee;Lee, Dae Bong;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.3
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    • pp.342-349
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    • 2018
  • In this study, the physicochemical properties, emulsifying capacity, moisture content and cytotoxicity of the composite material produced by transesterification reactions of the olive oil (olive oil esters) were investigated for cosmetic applications. Olive oil esters with short (S) and long (L) reaction times were studied. From the TLC-image analysis, composition ratios of the olive oil esters S were found to be 5.2, 24.1, 46.4, and 21.9% for mono-, di-, tri-glyceride, and fatty acid ethyl ester, respectively. Those of the olive oil esters L were 4.1, 24.7, 40.6, and 28.8% for mono-, di-, tri-glyceride, and fatty acid ethyl ester, respectively. The iodine value, acid value, saponification value, unsaponified matter, refractive index, and specific gravity were determined and purity tests were also carried out and normalized to establish standards and testing methods for using olive oil esters in cosmetics. To evaluate their emulsifying capacities, the O/W emulsion was prepared without surfactants and the formation of the emulsified particles were confirmed. After 5 days of applying the olive oil esters to human skin, the skin moisture retention was improved by 13.1% from the initial state. For the evaluation of toxicity on human skin cells, the olive oil esters showed 90% or more of the cell viability at $0.2-200{\mu}g/mL$. These results suggested that olive oil esters can be applied as natural/non-toxic ingredients to cosmetics industries.

Results of Three-Dimensional Conformal Radiation Therapy for the Treatment of a Solitary Sternal Relapse of Breast Cancer (흉골에 단독 전이된 유방암의 삼차원 입체조형 방사선 치료 성적)

  • Kim, Hae-Young;Huh, Seung-Jae;Park, Won;Choi, Do-Ho;Kang, Min-Kyu;Yang, Jung-Hyun;Nam, Seok-Jin;Im, Young-Hyuck
    • Radiation Oncology Journal
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    • v.26 no.2
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    • pp.91-95
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    • 2008
  • Purpose: To evaluate the response and survival rate after three-dimensional conformal radiation therapy(3D-CRT) of patients with a solitary sternal relapse of breast cancer. Materials and Methods: Seventeen patients between May 1996 and June 2005 were evaluated with the salvage 3D-CRT treatment of a solitary sternal relapse of breast cancer. The treatment fields included the gross tumor volume with 2 cm margins. The total radiation dose was $35.0{\sim}61.5$ Gy(biologic effective dose of $43.7{\sim}76.9Gy_{10}$ using an $\alpha/\beta$ ratio of 10 Gy), with a daily dose of $1.8{\sim}3.0$ Gy. The tumor response was evaluated by the change in maximum tumor size via follow up CT scans $1{\sim}3$ months after the completion of treatment. Results: An objective tumor response was achieved in all patients, with a complete response in 5 patients and a partial response in 12 patients. The 5-year overall survival rate was 51.9%(median survival time: 27 months), and the most important factor affecting overall survival was the disease-free interval(interval from primary surgery of breast cancer to the development of sternal metastasis): The 5-year overall survival rate was 61.8% for patients with a disease-free interval ${\geq}12$ months and 0.0% for patients with disease-free interval <12 months(p=0.03). Conclusion: The response to 3D-CRT was good in patients with solitary sternal relapse of breast cancer. Particularly, patients with long disease-free interval from primary surgery survived significantly longer than patients with short disease-free interval from primary surgery.