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Production of hTPO Transgenic Chickens using Tetracycline-Inducible Expression System (Tetracycline-Inducible Expression System을 이용한 Human Thrombopoietin (hTPO) 형질전환 닭의 생산)

  • Kwon, M.S.;Koo, B.C.;Kim, D.H.;Kim, M.J.;Kim, T.
    • Korean Journal of Poultry Science
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    • v.36 no.2
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    • pp.177-186
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    • 2009
  • It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

Effects of BCG on Gastric Chief Cells of the Mouse Implanted with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐 위점막 으뜸세포의 미세구조에 미치는 영향)

  • Ryoo, In-Sang;Ahn, E-Tay;Park, Kyung-Ho;Park, Dae-Kyoon;Kim, Myeong-Soo;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.35 no.3
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    • pp.153-163
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    • 2005
  • This experiment was performed to evaluate the morphological responses of the gastric chief cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG (Bacillus Calmette-Guerin). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with $1x10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the last injection, each mouse was sacrificed. Pieces of the tissue were taken from the stomach, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. The size of zymogen granule and the size of the mitochondrion of the gastric chief cells were observed and calculated. In the BCG treated group, most chief cells did not show any difference in ultrastructure, except that myelin figures were more frequently observed, in comparison with that of nornmal control group. The size of zymogen granule in the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.98({\pm}0.108){\mu}m,\;1.05({\pm}0.092){\mu}m\;and\;0.93({\pm}0.053){\mu}m$, respectively. And the mitochondrial size of the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.80({\pm}0.130){\mu}m,\;0.83({\pm}0.143){\mu}m\;and\;0.72({\pm}0.078){\mu}m$, respectively. From the above results, it was concluded that BCG may slightly suppress function of the gastric chief cells.

In Vitro Maturation of Tiger Oocytes : A Case Report (호랑이 난자의 체외성숙)

  • Lee, H.-S.;Yin, X.-J.;Lee, Y.-H.;Min, W.-K.;Kim, T.-S.;Choi, J.-W.;Yoon, B.-C.;Kim, J.-I.;Kong, I.-K.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.185-189
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    • 2004
  • The purpose of this study was to determine the possibility of in vitro maturation of tiger oocytes. Immature oocytes were recovered from a pair of ovaries. A total of 78 oocytes was collected, of which forty three were classified as good oocytes with compact cumulus cells and uniform cytoplasm. Forty three COCs were in vitro matured at $39^{\circ}C$, 5% CO2 in air atmosphere for 48 h in a IVM medium (TCM-199 supplement with 10% FBS, 0.6 mM cysteine, 0.2 mM pyruvic acid and 10 IU/mL HMG). Experiment I: the morphologic evaluation was conducted by measuring the diameter of oocytes with or without ZP, the thickness of ZP and the diameter of cytoplasm by microeyepiece at the same magnification (${\times}$100). Experiment II: the evaluation of meiotic development was conducted of the nuclear development stage of tiger oocytes. The results were summarized as follows: 1. The diameter of tiger oocytes $(176.5\pm6.1{\mu}m)$ with ZP was significantly (p<0.05) bigger than that of bovine oocytes $(150.7\pm4.9{\mu}m).$ The ZP thickness of tiger oocytes $(20.4\pm2.9{\mu}m)$ was significantly (p<0.05) bigger than that of bovine oocytes $(12.0\pm2.6{\mu}m;$ p<0.05). However, there was no significant difference in the diameter of cytoplasm (without ZP) between tiger $(122.1\pm9.7{\mu}m)$ and bovine oocytes $(118.7\pm7.5{\mu}m).$ 2. The rates of meiotic development of tiger oocytes were achieved GV (12.5 %) and MII (50.0%), respectively. These results indicated that tiger oocytes could be developed to MII in in vitro culture system.

Reliability of Muscle Evaluation with a Tactile Sensor System (촉각센서를 이용한 근육평가의 신뢰도 조사)

  • Oh, Young-Rak;Lee, Dong-Ju;Kim, Sung-Hwan;Kim, Mee-Eun;Kim, Ki-Suk
    • Journal of Oral Medicine and Pain
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    • v.30 no.3
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    • pp.337-344
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    • 2005
  • A tactile sensor employs a piezoelectric element to detect contact frequency shifts and thereby measure the stiffness or softness of material such as tissue, which allows the sensor to be used in many fields of research for urology, cardiology, gynecology, sports medicine and caner detection and especially for cosmetics and skin care. In this study, reliability of the tactile sensor system was investigated with its manual application to the muscles susceptible to temporomandibular disorders. Stiffness and elasticity of anterior temporalis, masseter and trapezius muscles were calibrated bilaterally from 5 healthy men with an average of 24.5$\pm$0.94 years. The tactile sensor used in this study had a computer-controlled and motor-driven sensor unit which automatically pressed down on the skin surface over the muscles being measured and retracted, thereby providing the hysteresis curve. The slope of the tangent of the hysteresis curve (${\Delta}f/{\Delta}x$) is defined as stiffness of the muscle being measured and the distance between the two parts of the curve as its elasticity. To determine inter-examiner reliability, all the measurements were performed by the two examiners A and B, respectively and the same examination were repeated with an interval of 2 days for intra-examiner reliability. The results from this study demonstrated high reliability in measuring stiffness and elasticity of anterior temporalis, masseter and upper trapezius muscles using a tactile sensor system. It is suggested that the tactile sensor system can be a highly reproducible and effective instrument for quantitative evaluation of the muscle in head and neck region.

Studies on the Generation-to-Generation Transmission of Cytoplasmic Polyhedrosis Virus and the Effect of Their Activation on the Induction in the Next Generation in the Silkworm, Bombyx mori L. (Virus의 경란전염이 차대의 잠작에 미치는 영향에 관한 연구)

  • 임종성;김근영
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.85-92
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    • 1974
  • Many of studies on the transovarial transmission of occult virus and their activation due to various stresses such as cold or heat treatment, chemical feeding, and nutritional deficiency, etc., in the silkworm, Bombyx mori L. have been made, but any attempts have been not made to control virus diseases by detection of the occult virus-carried moths in the production of silkworm egg of hybrids, because of difficulty to detect occult virus in any stage. Therefore, it may be worth while to disclose whether a sublethal infection of the moths from which active virus are detectable, has the same level of induction rate as that of occult virus activation, thus to apply its results for the reduction of the occurence of virus diseases in silkworm rearing. For these purposes, the following experiment was conducted as one of preliminary steps. In this study, investigations on the generation-to-generation transmission of occult virus and a sublethal infection, and the role of chromosomal gene of the host, Jam 103 and Jam 104 in the Previous generation, and Jam 103 x Jam 103 and Jam 104 f Jam 104 in the next generation were made for the induction of virus diseases due to the transmitted virus. The frequency of cytoplasmic polyhedrosis due to the induction in the F$_1$ generation was markedly higher in the cross-batches, male$\times$female and male$\times$female in which inoculated individuals were used as fem ale parents than in the cross-batches, male$\times$female and male$\times$female in which virus has been not inoculated or inoculated only to male in the previous generation. The tendency of increasing rate was observed in any treatments; such as the inoculations of cytoplasmic polyhedrosis virus (10$\^$5/, 10$\^$6/ 10$\^$7, and 10$\^$8//ml ill different concentration of inocula) , cold-treatment (5$^{\circ}C$, 12hrs or 24hrs), and formalin-feeding treatment (2% or 3%). The shape of polyhedra (tetragonal in outline) examined in the F, larvae was identified as that of the inoculated polyhedra with partial application of immunofluorescent techniques. These results suggests that the cytoplasmic polyhedrosis virus in B. meri L. are transmitted to the next generation through the egg, apparently in the occult state. And the experimental results of various cross-batches revealed the egg cytoplasm plays an important part i the transmission of the occult virus of the cytoplasmic polyhedrosis virus,

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Effect of Dietary Fish Oil on Lipid Peroxidation in Rats Liver and Brain During Postnatal Development (어유섭취가 출생후 발달과정의 흰 쥐의 간과 뇌조직의 지질과산화와 그 관련기능에 미치는 영향)

  • 박명희
    • Journal of Nutrition and Health
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    • v.20 no.2
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    • pp.111-121
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    • 1987
  • Lipie peroxide formation, antiperoxidative s system and body adaptability for handling lipid p peroxide were examined in the first and second g generations of rats fed fish oil. Mackerel oil(MO) was used and four other dietary oils and fat, i.e. soybean oil(SO), perilla oil(PO), rapeseed oil(RO) and beef tallow(BT) were also employed to compare the effect of fish oil. Synthetic diets containing these five dietary fats at the level of 1O%(w/w), were given to the correspond­m ing groups of male and female rats weighing about 70 grams. After 34 days of feeding, male a and female rats were mated and their offsprings were raised throughout suckling (17, 26 days) and weanling (39 days) periods. Liver lipid pero­x xide level was highest in MO group of both first (mother rats after lactation) and second genera­t tions of 17 and 26 days old, but not of 39 days old. During suckling period, liver lipid peroxide level was well matched to total unsaturation of dietary fat. Brain lipid peroxide levels were not different among five groups. Liver $alpha$-tocopherol a and reduced glutathione (GSH) levels were lowest in MO fed first generation. In second generation, $alpha$-tocopherol level was also low in MO group, although the effect was less pronoun­c ced, but GSH level was not different from other groups. Oxidized glutathione (GSSG) level did not consistently vary by change in dietary fat. Glutathione peroxidase activity increased as young rats grew up to 39 days. Superoxide d dismutase activity change was insignificant by a age, but was shown as lowest in MO group. At the age of 26 and 39 days, liver glutatione peroxidase activity was increased as was level of lipid peroxide, suggesting that this is the one of the mechanisms responsible for body adapta­b bility for protection against the accumulation of lipid peroxide.

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Active Aging: Roles of Physical Activity and Immunity (건강한 노후 : 운동활동과 면역반응을 중심으로)

  • Park, Chan Ho;Kim, Ji-Seok;Kwak, Yi Sub
    • Journal of Life Science
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    • v.28 no.5
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    • pp.621-626
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    • 2018
  • We introduced the physiological responses of aging, active aging and also suggest the impact of physical exercise on body health status and elderly immunity. In this purpose, we searched the Pub Med data base for the articles (include our experimental papers) and review papers having the terms 'Aging', 'Active aging' and 'Physical activity and elderly' in the title, published from 1999 until 2018. The results were as follows: Exercise training has been extensively studied about the reduction of inflammation, oxidative stress, disease, and aging in syndrome X patients and elderly. Combined and aerobic or resistance exercise training could reduce obesity, insulin resistance, type 2 diabetes and hypertension. Exercise training has been extensively studied in cancer settings as part of prevention or treatment strategies. From this research, regular exercise has the potential to target tumor growth through regulation of inflammation and immune responses such as lactate clearance, NK cell activation (innate immunity), activation of cytotoxic immune cells, T cell activation (adaptive immunity), and immune surveillance. However, Endurance physical activity not only induces thermogenesis and diverse sports injuries but also elicits mobilization and functional enhancement of monocytes, neutrophils (which is caused by the cytokine changes such as TNF-alpha, IL-1) whereas it suppresses cell mediated immunity causing to increased susceptibility to inflammation and infections like cough and URTIs (upper respiratory track infections) in young and especially in elderly people. Therefore, Strategies to prevent physical fatigue, sports injuries include avoid overtraining, Adequate recovery and various type of rest during and after physical activity and assuring adequate nutrition supplementation such as glutamine, vitamin B, vitamin C, carbohydrate, ion or berry-contain sports beverages is helpful in physically active elderly.

A Study on the IADL Affecting Subjective Health Index of the Aged in Some Area (일부지역(一部地域) 노인(老人)들의 주체적(主體的) 건강수준(健康水準)에 영향을 미치는 IADL에 관한 연구(硏究))

  • Kim, Keun-Jo;Park, Heung-Ki;Koon, Hyeok-Su;Bae, Soo-Chan
    • The Journal of Korean Academy of Orthopedic Manual Physical Therapy
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    • v.7 no.1
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    • pp.29-50
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    • 2001
  • This research have been made to define how the IADL (Instrumental Activity of Daily Living) performance and the subjective health index of the Aged are affected by their residential circumstance, gender and age, and how deeply these two factors are related and interact. For the period of June 1 to July 31, 2000, we had conducted a questionnaire and direct interview with 693 persons over age sixty-five (65) in Daejon and in other adjacent area, grouping into three different residential types The Aged living at home, The Aged living at welfare facilities and The Aged living alone, and studied on how the IADL performance and the subjective health index of the Aged are influenced and interact as per their characteristics, daily activity and mentality. We had analyzed all the data obtained through this research by the method of : - analysis of frequency as per specific factors by SPSS 1O.0/PC+, - $x^2$ test, - t-test, - ANOVA, - multiple regression analysis by factors. The research concludes followings : a. It appears that the three (3) factors such as gender, age and residential circumstance of the Aged deeply affect the IADL performance and subjective health index of the Aged. (p<0.01) b. With regard to IADL performance of the Aged by the gender, it was analyzed that the female-Aged gains 23.8 point on average, which shows the performance of the female-Aged is less independent. (p<0.01). In addition, it was also found that the IADL performance is becoming less and less independent following their age increasing. In analyzing IADL performance by the residential type, it appears that the Aged living at welfare facilities gains the lowest 21.5 points and is least independent. It was also found that the Aged living at welfare facilities need some assistances from others for their performing IADL. (20-24 point) (p<0.001) c. With regard to the subjective health index of the all-Aged participated in this research, the analysis indicates 8.8 point and this is considered as a point of general standard (7-10 point). In analyzing this index by the gender, the female-Aged gains only 8.6 point which explains a lot of female-Aged consider they are not really healthy. (p<0.001) In analyzing this index by the residential type, the Aged living at welfare facilities and the Aged living alone gain the comparatively lower point, - respectively 8.4 point for the Aged living at welfare facilities and 8.8 point for the Aged living alone. The Aged living at these two residential types express they are obviously in a bad situation of health. (p<0.001) d. With regard to the factors affecting the IADL performance and the subjective health index of the Aged, it was analyzed that the IADL performance can largely be affected by the factors such as depression, frequency of going-out and age rising, and that the subject health index can also deeply be affected by depression, pain and by how much they are satisfied with their current living conditions. e. It was analyzed that the interacting between the IADL performance and the subjective health index is not that strong but even weak. As a result, we were able to conclude that the IADL performance is less independent in case of the female-Aged, the Aged living at welfare facilities, and following the age rising. As for the subjective health state the Aged themselves are aware of, we concluded that the female-Aged, the Aged living at welfare facilities and the Aged living alone, are more critical about their health. From this research, we were able to realize that, when the OMT (Orthopedic Manual Therapy) needed, the physical therapists are really required to have a correct and cautious understanding of the situation in which the aged persons are, and take care of them with more concerns and more improved treatment.

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Studies on the ecological variations of rice plant under the different seasonal cultures -II. A study on the year variations and prediction of heading dates of paddy rice under the different seasonal cultures- (재배시기 이동에 의한 수도의 생태변이에 관한 연구 -II. 재배시기 이동에 의한 수도출수기의 년차간변이와 그 조기예측-)

  • Hyun-Ok Choi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.3
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    • pp.41-48
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    • 1965
  • This study was aimed at knowing the magnitude of year variation in rice heading dates under the different seasonal cultures, and to estimate the heading date in advance. Using six rice varieties such as Kwansan, Suwon#82, Suwon #144, Norin#17, Yukoo#132 and Paltal, the early, ordinary and late seasonal cultures had been carried out at Paddy Crop Division, Crop Experiment Station at Suwon for the six-year period 1959 to 1964. In addition the data of the standard rice cultures at the Provincial Offices of Rural Development for the 12-year period 1953 to 1954, were analyzed for the purpose of clarifying a relationship between variation of rice heading dates and some of meteorological data related to the locations and years. The results of this study are as follows: 1. Year variation of rice heading dates was as high as 14 to 21 days in the early seasonal culture and 7 to 14 days in the ordinary seasonal culture, while as low as one to seven days in the late seasonal culture which was the lowest among three cultures. The magnitude of variation depended greatly on variety, cultural season and location. 2. It was found out that there was a close negative correlation between the accumulated average air temperature for 40 days from 31 days after seeding and number of days to heading in the early seasonal culture. Accordingly, it was considered possible to predict the rice heading date through calculation of the accumulated average air temperature for the above period and then the linear regression(Y=a+bx). On the other hand, an estimation of the heading date in the late seasonal culture requires for the further studies. In the ordinary seasonal culture, no significant correlation between the accumulated average air temperature and number of days to heading was obtained in the six-year experiments conducted at Suwon. There was a varietal difference in relationship between the accumulated average air temperature for 70 days from seeding and number of days to heading in the standard cultures at the provincial offices of rural development. Some of varieties showed a significant correlation between two factors while the others didn't show any significant correlation. However, there was no regional difference in this relationship.

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Relationship between XRCC1 Polymorphism and Acute Complication of Chemoradiation Therapy in the Patients with Colorectal Cancer (대장, 직장암 환자에서 화학방사선치료의 급성 부작용과 XRCC1 유전자 다형성과의 상관관계)

  • Kim Woo-Chul;Hong Yun-Chul;Choi Sun-Keun;Woo Ze-Hong;Nam Jeong-Hyun;Choi Gwang-Seong;Lee Moon-Hee;Kim Soon-Ki;Song Sun-U.;Loh John-Jk
    • Radiation Oncology Journal
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    • v.24 no.1
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    • pp.30-36
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    • 2006
  • Purpose: It is well known from clinical experience that acute complications of chemoradiation therapy vary from patients to patients. However, there are no known factors to predict these acute complications before treatment starts. The human XRCC1 gene is known as a DNA base excision repair gene. We investigated the possibilities of XRCC1 gene polymorphisms as a predictor for the acute complications of chemoradiation therapy in colorectal cancer patients. Materials and Methods: From July 1997 to June 2003, 86 colorectal cancer patients (71 rectal cancer, 13 sigmoid colon cancer and 2 colon cancer patients) were treated with chemoradiation therapy at the Department of Radiation Oncology, Inha University Hospital. Twenty-two patients were in stage B, 50 were in stage C, 8 were in stage D and 6 patients were unresectable cases. External radiation therapy was delivered with 10MV X-ray at a 1.8 Gy fraction per day for a total dose of radiation of $30.6{\sim}59.4 Gy$ (median: 54 Gy). All the patients received 5-FU based chemotherapy regimen. We analyzed the acute complications of upper and lower gastrointestinal tract based on the RTOG complication scale. The initial and lowest WBC and platelet count were recorded during both the RT period and the whole treatment period. Allelic variants of the XRCC1 gene at codons 194, 280 and 399 were analyzed in the lymphocyte DNA by performing PCR-RFLP. Statistical analyses were carried out with the SAS (version 6.12) statistical package. Results: When all the variables were assessed on the multivariate analysis, recurrent disease revealed the factors that significantly correlated with upper gastrointestinal acute complications. Arg399Gln polymorph isms of the XRCC1 gene, the radiation dose and the frequencies of chemotherapy during radiation therapy were significantly correlated with lower gastrointestinal complications. Arg399Gln polymorph isms also affected the decrease of the WBC and platelet count during radiation therapy. Conclusion: Although the present sample size was too small for fully evaluating this hypothesis, this study suggests that Arg399Gln polymorph isms of the XRCC1 genes may be used as one of the predictors for acute complications of chemoradiation therapy in colorectal cancer patients.