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The Effects of 8-week Ketone Body Supplementation on Endurance Exercise Performance and Autophagy in the Skeletal Muscle of Mice (8주 케톤체 투여가 마우스 지구성 운동수행능력과 골격근의 자가포식에 미치는 영향)

  • Jeong-sun Ju;Min-joo Park;Dal-woo Lee;Dong-won Lee
    • Journal of Life Science
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    • v.33 no.3
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    • pp.242-251
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    • 2023
  • The purpose of this study was to investigate the effects of 8-week β-hydroxybutyrate (β-HB) administration with and without endurance exercise training on endurance exercise performance and skeletal muscle protein synthesis and degradation using a mouse model. Forty-eight male wild-type ICR mice (8 weeks old) were randomly divided into four groups: sedentary control (Sed+Con), (Sed+Con), sedentary β-HB (Sed+β-HB), exercise control (Exe+Con), and exercise β-HB (Exe+β-HB). β-HB was dissolved in PBS (150 mg/ml) and injected subcutaneously daily (250 mg/kg) for 8 weeks. Mice performed 5 days/week of a 20 min treadmill running exercise for 8 weeks. The running exercise was carried out at a speed of 10 m/min at a 10° incline for 5 min, and then the speed was increased by 1 m/min for every 1 min of the remaining 15 min. Following 8 weeks of treatments, visceral fat mass and skeletal muscle mass, blood parameters, and the markers for autophagy and protein synthesis were analyzed. The data were analyzed with one-way ANOVA (p<0.05) using the SPSS 21 program. Eight weeks of Exe+β-HB treatment significantly lowered blood lactate levels compared with the other three groups (Sed+Con, Sed+β-HB, and Exe+β-HB) Exe+β-HB) (p<0.05). Eight weeks of Exe+β-HB significantly increased maximal running time (time to exhaustion) compared with the Sed+Con and Exe+Con groups (p<0.05). Eight weeks of β-HB administration significantly decreased autophagy flux and autophagy-related proteins in the skeletal muscle of mice (p<0.05). Conversely, the combined treatment of β-HB and endurance exercise training increased protein synthesis (mTOR signaling and translation) (p<0.05). The 8-week β-HB treatment and endurance exercise training had synergistic effects on the increase in endurance performance, increase in protein synthesis, and decrease in protein degradation in the skeletal muscle of mice.

Changes in chemical composition and physiological activity of Jeju-Tatary buckwheat tea according to leaching temperature (제주 타타리메밀의 침출 조건에 따른 제주 타타리메밀침출차의 이화학적 특성 및 생리활성)

  • Hyun-A Ko;Hyun Ju Park;Inhae Kang
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.421-427
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    • 2022
  • In this study, Jeju Tatary buckwheat tea's chemical composition and physiological activities were compared according to the leaching temperature (60, 80, 100 ℃). As the leaching temperature is increased, the degree of browning is induced. However, there was no significant change in pH. The total polyphenol content was higher at 80 ℃ than at 60 ℃ leaching temperature, but significantly decreased at 100 ℃ leaching temperature (60 ℃: 17.06 mg GA/g, 80 ℃: 20.09 mg GA/g, 100 ℃ :18.45 mg GA/g). There were high content of flavonoid and rutin as the leaching temperature increased. Consistently, 2,2-diphenyl1-picrylhydrazyl (DPPH) radical scavenging activity and tyrosinase inhibitory activity were significantly higher with increasing temperature (DPPH % inhibition: 60 ℃: 41.88%, 80 ℃: 46.01%, 100 ℃: 46.80%/tyrosinase inhibitory activity: 60 ℃: 9.38%, 80 ℃: 22.94%, 100 ℃: 28.17%). However, there was no significant difference in DPPH radical scavenging activity between 80 and 100 ℃. A cytotoxicity test was performed by treating with Jeju Tatary buckwheat extract into mouse macrophage cells (Raw264.7). 100 and 200 ㎍/mL treatment (100 ℃ extract) were significantly upregulated the survival rate, but there was no significant difference in other concentrations. Collectively, most of the bioactive components, antioxidant activity, and tyrosinase inhibitory activity were induced as the leaching temperature increased. However, the content of polyphenols which are known to have antioxidant activity, was significantly reduced at 100 ℃ leaching temperature. Several reports have demonstrated that leaching at too high temperature lowered the overall acceptability, so the optimal leaching condition of Tatary Buckwheat is 80 ℃, 5 min in this study.

Development of High Intensity Focused Ultrasound (HIFU) Mediated AuNP-liposomal Nanomedicine and Evaluation with PET Imaging

  • Ji Yoon Kim;Un Chul Shin;Ji Yong Park;Ran Ji Yoo;Soeku Bae;Tae Hyeon Choi;Kyuwan Kim;Young Chan Ann;Jin Sil Kim;Yu Jin Shin;Hokyu Lee;Yong Jin Lee;Kyo Chul Lee;Suhng Wook Kim;Yun-Sang Lee
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.9 no.1
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    • pp.9-16
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    • 2023
  • Liposomes as drug delivery system have proved useful carrier for various disease, including cancer. In addition, perfluorocarbon cored microbubbles are utilized in conjunction with high-intensity focused-ultrasound (HIFU) to enable simultaneous diagnosis and treatment. However, microbubbles generally exhibit lower drug loading efficiency, so the need for the development of a novel liposome-based drug delivery material that can efficiently load and deliver drugs to targeted areas via HIFU. This study aims to develop a liposome-based drug delivery material by introducing a substance that can burst liposomes using ultrasound energy and confirm the ability to target tumors using PET imaging. Liposomes (Lipo-DOX, Lipo-DOX-Au, Lipo-DOX-Au-RGD) were synthesized with gold nanoparticles using an avidin-biotin bond, and doxorubicin was mounted inside by pH gradient method. The size distribution was measured by DLS, and encapsulation efficiency of doxorubicin was analyzed by UV-vis spectrometer. The target specificity and cytotoxicity of liposomes were assessed in vitro by glioblastoma U87mg cells to HIFU treatment and analyzed using CCK-8 assay, and fluorescence microscopy at 6-hour intervals for up to 24 hours. For the in vivo study, U87mg model mouse were injected intravenously with 1.48 MBq of 64Cu-labeled Lipo-DOX-Au and Lipo-DOX-Au-RGD, and PET images were taken at 0, 2, 4, 8, and 24 hours. As a result, the size of liposomes was 108.3 ± 5.0 nm at Lipo-DOX-Au and 94.1 ± 12.2 nm at Lipo-DOX-Au-RGD, and it was observed that doxorubicin was mounted inside the liposome up to 52%. After 6 hours of HIFU treatment, the viability of U87mg cells treated with Lipo-DOX-Au decreased by around 20% compared to Lipo-DOX, and Lipo-DOX-Au-RGD had a higher uptake rate than Lipo-DOX. In vivo study using PET images, it was confirmed that 64Cu-Lipo-DOX-Au-RGD was taken up into the tumor immediately after injection and maintained for up to 4 hours. In this study, drugs released from liposomes-gold nanoparticles via ultrasound and RGD targeting were confirmed by non-invasive imaging. In cell-level experiments, HIFU treatment of gold nanoparticle-coupled liposomes significantly decreased tumor survival, while RGD-liposomes exhibited high tumor targeting and rapid release in vivo imaging. It is expected that the combination of these models with ultrasound is served as an effective drug delivery material with therapeutic outcomes.

Molecular Breeding of Tobacco Plants Resistant to TMV and PVY (분자생물학적 TMV 및 PVY 저항성 연초 육종)

  • E.K. Pank;Kim, Y.H.;Kim, S.S.;Park, S.W.;Lee, C.H.;K.H.Paik
    • Proceedings of the Korean Society of Tobacco Science Conference
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    • 1997.10a
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    • pp.134-152
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    • 1997
  • Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

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L-AHG-mediated Suppression of M1 Polarization and Pro-inflammatory Signaling Pathways in LPS-stimulated RAW264.7 Macrophages (LPS에 의해 자극된 RAW264.7 대식세포에서 L-AHG에 의한 M1 분극화 및 친염증 신호 경로의 억제)

  • Won Young Jang;Shin Young Park;Ki Youn Kim;Do Youn Jun;Young-Seuk Bae;Young Ho Kim
    • Journal of Life Science
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    • v.34 no.7
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    • pp.443-452
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    • 2024
  • This study aimed to examine the influence of 3,6-anhydroxygalactose (L-AHG) on the pro-inflammatory M1 polarization and pro-inflammatory responses observed in the RAW264.7 mouse macrophage cell line following stimulation with lipopolysaccharides (LPS). L-AHG exhibited a significant and dose-dependent inhibition of inducible nitric oxide synthase (iNOS) expression, a hallmark of M1 polarization, and subsequent NO production in LPS-stimulated RAW264.7 cells. Furthermore, the LPS-induced upregulation of cyclooxygenase-2 (COX-2), which drives the production of prostaglandin E2, an inflammatory mediator, was also inhibited by L-AHG. L-AHG did not affect the LPS-triggered Toll-like receptor 4 (TLR4)-mediated pro-inflammatory signaling pathway, which culminated in the activation of transforming growth factor-β-activated kinase 1 (TAK1). However, it was observed to inhibit the generation of reactive oxugen species (ROS) in a dose-dependent manner, as well as the TAK1-driven activation of JNK and p38 MAPK. Given that the active p38 MAPK is known to contribute to the assembly of active nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, which catalyzes the intracellular generation of pro-inflammatory ROS in LPS-stimulated macrophages, the dose-dependent reduction in the LPS-induced ROS generation by L-AHG may be mainly due to the prevention of TAK1-driven activation of p38 MAPK. Together, these results demonstrate that the L-AHG-mediated inhibition of the TAK1-JNK/p38 MAPK activation phase of the pro-inflammatory signaling pathway in LPS-stimulated RAW264.7 cells by L-AHG represents a promising mechanism for suppressing M1 polarization and pro-inflammatory responses in macrophages.

Study on skin anti-inflammatory activity of fig (Ficus carica L.) fruit extract fractions (무화과(Ficus carica L.) 열매 추출 분획의 피부 항염증 활성 연구)

  • Hee Joon Kwon;Geun soo Lee;Jin Hwa Kim;Soon Woo Kwon;Hyung seo Hwang
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.416-423
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    • 2023
  • Figs has known to have antioxidant, whitening, anti-inflammatory, and antibacterial effects in their leaves, roots, stems, latex, and fruits. In order to develop cosmetic materials based on natural products, we have studied on the skin activity of the ficin in latex as well as the whitening function of the fruit extract with 70% ethanol, and used it as a raw material for released cosmetic product. However, there is little research on the demand for the development of new eutectic solvent extraction methods and its ability to control skin inflammation and psoriasis regulation. Thus, in this study, we evaluated the effectiveness of fig fruit extracts and fractions using eutectic solvent extraction for skin inflammation control and psoriasis. First, fig fruits were extracted under optimal eutectic solvent conditions and fractionated with n-hexane, dichloromethane, ethyl acetate, and butanol. First, the antioxidant activity and inhibition of nitric oxide (NO) production were confirmed in mouse macrophage RAW264.7 cells. In addition, as a result of observing the mRNA expression through RT-PCR, pro-inflammatory cytokines such as TNF-α, IL1α, and IL-1β were suppressed significantly in the hexane, dichloromethane, and ethyl acetate fractions. In addition, it was confirmed in TNF-α stimulated HaCaT keratinocyte model. Finally, chemokine CC motif ligand 20 (CCL20), marker gene of human psoriasis skin disease, was significantly suppressed in the hexane, dichloromethane, and ethyl acetate fractions. These results suggested its anti-inflammatory and skin soothing effect and the possibility of development as an excellent skin soothing natural cosmetic material in the future through future clinical trials.

The Effects of Salinity Stress on the Antioxidant and Anti-inflammatory Activities of Crepidiastrum sonchifolium (염분 스트레스가 고들빼기의 항산화 및 항염증 활성에 미치는 영향)

  • Ha Young Baek;Yeong Geun Song;Kyungjun Kim;Hyungjoo Kim;Kyeong Cheol Lee;Cheol-Joo Chae;Hyun Jung Koo
    • Journal of Practical Agriculture & Fisheries Research
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    • v.26 no.2
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    • pp.5-13
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    • 2024
  • This study was conducted to investigate the effect of salinity stress on the antioxidant and anti-inflammatory activities of Crepidiastrum sonchifolium. The plant was treated with NaCl at concentrations of 0, 50, 100, and 200mM for 6 weeks. After treatment, the whole plant was collected, and 70% ethanol extracts were prepared. The DPPH radical scavenging activity was highest in the order of NaCl treatment concentrations of 0, 100, and 50mM, while the 200mM treatment group showed the lowest radical scavenging. The total phenol and total flavonoid contents showed very similar results to the antioxidant activity depending on the NaCl concentration, confirming that the phenolic compounds of the plant can contribute to the antioxidant capacity against salinity stress. In addition, the investigation of the effect of NaCl-treated C. sonchifolium extract on the inhibition of NO production in the LPS-stimulated mouse macrophage cell line (Raw 264.7) revealed that NO production significantly decreased in the 1,000㎍/mL treatment group across all NaCl concentration groups. But, the high concentration (1,000㎍/mL) treatment of the 100mM and 200mM NaCl treatment groups was found to have a negative effect on cell survival. These results suggest that radical scavenging activity is highest in healthy plants and that they produce antioxidants to respond to NaCl salinity stress up to 100mM. However, a high NaCl concentration of 200mM has a negative effect on the physiological activity of the plants. Compared with the results of the previously reported growth index, it is thought that the growth and physiological activity of plants can be positively affected in an NaCl treatment environment of 50mM or less.

Detoxification of PSP and relationship between PSP toxicity and Protogonyaulax sp. (마비성패류독의 제독방법 및 패류독성과 원인플랑크톤과의 관계에 관한 연구)

  • CHANG Dong-Suck;SHIN Il-Shik;KIM Ji-Hoe;PYUN Jae-hueung;CHOE Wi-Kung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.22 no.4
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    • pp.177-188
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    • 1989
  • The purpose of this study was to investigate the detoxifying effect on PSP-infested sea mussel, Mytilus edulis, by heating treatment and correlation between the PSP toxicity and the environmental conditions of shellfish culture area such as temperature, pH, salinity, density of Protogonyaulax sp. and concentration of inorganic nutrients such as $NH_4-N,\;NO_3-N,\;NO_2-N\;and\;PO_4-P$. This experiment was carried out at $Suj\u{o}ng$ in Masan, Yangdo in Jindong, $Hach\u{o}ng\;in\;K\u{o}jedo\;and\;Gamch\u{o}n$ bay in Pusan from February to June in $1987\~1989$. It was observed that the detection ratio and toxicity of PSP in sea mussel were different by the year even same collected area. The PSP was often detected when the temperature of sea water about $8.0\~14.0^{\circ}C$. Sometimes the PSP fox of sea mussel was closely related to density of Protogonyaulax sp. at $Gamch\u{o}n$ bay in Pusan from March to April in 1989, but no relationship was observed except above duration during the study period. The concentration of inorganic nutrients effects on the growth of Protogonyaulax sp., then effects of $NO_3-N$ was the strongest among them. When the PSP-infested sea mussel homogenate was heated at various temperature, the PSP toxicity was not changed significantly at below $70^{\circ}C$ for 60 min. but it was proper-tionaly decreased as the heating temperature was increased. For example, when the sea mussel homogenate was heated at $100^{\circ}C,\;121^{\circ}C$ for 10 min., the toxicity was decreased about $67\%\;and\;90\%$, respectively. On the other hand, when shellstock sea mussel contained PSP of $150{\mu}g/100g$ was boiled at $100^{\circ}C$ for 30 min. with tap water, the toxicity was not detected by mouse assay, but that of PSP of $5400{\mu}g/100g$ was reduced to $57{\mu}g/100g$ even after boiling for 120 min.

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Standard Chemotherapy with Excluding Isoniazid in a Murine Model of Tuberculosis (마우스 결핵 모델에서 Isoniazid를 제외한 표준치료의 예비 연구)

  • Shim, Tae Sun;Lee, Eun Gae;Choi, Chang Min;Hong, Sang-Bum;Oh, Yeon-Mok;Lim, Chae-Man;Lee, Sang Do;Koh, Younsuck;Kim, Woo Sung;Kim, Dong Soon;Cho, Sang-Nae;Kim, Won Dong
    • Tuberculosis and Respiratory Diseases
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    • v.65 no.3
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    • pp.177-182
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    • 2008
  • Background: Isoniazid (INH, H) is a key drug of the standard first-line regimen for the treatment of tuberculosis (TB), yet some reports have suggested that treatment efficacy was maintained even though INH was omitted from the treatment regimen. Methods: One hundred forty C57BL/6 mice were infected with the H37Rv strain of M. tuberculosis with using a Glas-Col aerosol generation device, and this resulted in depositing about 100 bacilli in the lung. Four weeks after infection, anti-TB treatment was initiated with varying regimens for 4-8 weeks; Group 1: no treatment (control), Group 2 (4HREZ): 4 weeks of INH, rifampicin (R), pyrazinamide (Z) and ethambutol (E), Group 3: 1HREZ/3REZ, Group 4: 4REZ, Group 5: 4HREZ/4HRE, Group 6: 1HREZ/3REZ/4RE, and Group 7: 4REZ/4RE. The lungs and spleens were harvested at several time points until 28 weeks after infection, and the colony-forming unit (CFU) counts were determined. Results: The CFU counts increased steadily after infection in the control group. In the 4-week treatment groups (Group 2-4), even though the culture was negative at treatment completion, the bacilli grew again at the 12-week and 20-week time points after completion of treatment. In the 8-week treatment groups (Groups 5-7), the bacilli did not grow in the lung at 4 weeks after treatment initiation and thereafter. In the spleens of Group 7 in which INH was omitted from the treatment regimen, the culture was negative at 4-weeks after treatment initiation and thereafter. However, in Groups 5 and 6 in which INH was taken continuously or intermittently, the bacilli grew in the spleen at some time points after completion of treatment. Conclusion: TThe exclusion of INH from the standard first-line regimen did not affect the treatment outcome in a murine model of TB in the early stage of disease. Further studies using a murine model of chronic TB are necessary to clarify the role of INH in the standard first-line regimen for treating TB.

Radioprotective Effects of Granulocyte-Colony Stimulating Factor in the Jejunal Mucosa of Mouse (생쥐에서 과립구 집락형성인자(Granulocyte-Colony Stimulating Factor)의 공장점막에 대한 방사선 보호효과)

  • Ryu, Mi-Ryeong;Chung, Su-Mi;Kay, Chul-Seung;Kim, Yeon-Shil;Yoon, Sei-Chul
    • Radiation Oncology Journal
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    • v.19 no.1
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    • pp.45-52
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    • 2001
  • Purpose : Granulocyle-colony stimulating factor (G-CSF) has been widely used to treat neutropenia caused by chemotherapy or radiotherapy. The efficacy of recombinant human hematopoietic growth factors in improving oral mucositis after chemotherapy or radiotherapy has been recently demonstrated in some clinical studies. This study was designed to determine whether G-CSF can modify the radiation injury of the intestinal mucosa in mice. Materials and Methods : One hundred and five BALB/c mice weighing 20 grams were divided into nine subgroups including G-CSF alone group $(I:10\;{\mu}g/kg\;or\;II:100\;{\mu}g/kg)$, radiation alone group (7.5 or 12 Gy on the whole body), combination group with G-CSF and radiation (G-CSF I or II plus 7.5 Gy, G-CSF I or II plus 12 Gy), and control group. Radiation was administered with a 6 MV linear accelerator (Mevatron Siemens) with a dose rate of 3 Gy/min on day 0. G-CSF was injected subcutaneously for 3 days, once a day, from day -2 to day 0. Each group was sacrificed on the day 1, day 3, and day 7. The mucosal changes of jejunum were evaluated microscopically by crypt count per circumference, villi length, and histologic damage grading. Results : In both G-CSF I and II groups, crypt counts, villi length, and histologic damage scores were not significantly different from those of the control one (p>0.05). The 7.5 Gy and 12 Gy radiation alone groups showed significantly lower crypt counts and higher histologic damage scores compared with those of control one (p<0.05). The groups exposed to 7.5 Gy radiation plus G-CSF I or II showed significantly higher crypt counts and lower histologic damage scores on the day 3, and lower histologic damage scores on the day 7 compared with those of the 7.5 Gy radiation alone one (p<0.05). The 12 Gy radiation plus G-CSF I or II group did not show significant difference in crypt counts and histologic damage scores compared with those of the 12 Gy radiation alone one (p>0,05). Most of the mice in 12 Gy radiation with or without G-CSF group showed intestinal death within 5 days. Conclusion : These results suggest that G-CSF may protect the jejunal mucosa from the acute radiation damage following within the tolerable ranges of whole body irradiation in mice.

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