Immune Cell Activation and Co-X-irradiation Effect of Eleutherococcus senticosus Maxim Root / 대한방사선종양학회지

PURPOSE: This study was performed to investigate the effects of immune cell activation and the antitumor effect for the combination of treatment with X-irradiation and Eleutherococcus senticosus Maxim Root (ESMR) on mouse tumor cells. MATERIALS AND METHODS: ESMR (250g) was extracted with 80% methanol, concentrated under decompression and lyophilized. To determine whether ESMR is able to activate the immune cells or not, the proliferation of splenocytes in vitro and the number of B cells and T cells in splenic lymphocytes in ESMR-pretreated mice were evaluated. X-irradiation was given to the mouse fibrosarcoma tumor cells (FSa II) by 250 kv X-irradiation machine. The cytotoxicity of ESMR was evaluated from its ability to reduce the clonogenecity of FSa II cells. In X-irradiation alone group, each 2, 4, 6 and 8 Gy was given to FSa II cells. In X-irradiation with ESMR group, 0.2 mg/ml of ESMR was exposed to FSa II cells for 1 hour before X-irradiation. RESULTS: The proliferation of cultured mouse splenocytes and thymocytes were enhanced by the addition of ESMR in vitro. The number of B cells and T cells in mouse splenic lymphocytes was significantly increased in ESMR pretreated mice in vivo. In FSa II cells that received a combination of 0.2 mg/ml of ESMR with X-irradiation exposure, the survival fraction with a dose of 2, 4 and 6 Gy was 0.39+/-0.005, 0.22+/-0.005 and 0.06+/-0.007, respectively. For FSa II cells treated with X-irradiation alone, the survival fraction with a dose of 2, 4 and 6 Gy was 0.76+/-0.02, 0.47+/-0.008 and 0.37+/-0.01. The difference in the survival fraction of the mouse FSa II cells treated with and without ESMR was statistically significant (p<0.05). CONCLUSION: Treatment with ESMR increased cell viability of mouse splenocytes in vitro and especially the subpopulation of B cells and T cells in splenocytes in ESMR-pretreated mice. However, treatment with ESMR did not increase the level of Th and Tc subpopulations in the thymocytes. Treatment with the combination of ESMR and X-irradiation was more cytotoxic to mouse tumor cells than treatment with X-irradiation alone; this finding was statistically significant.

A Confocal Laser Scanning Microscopic Study on the Distribution and Fluorescent Intensity of NPY-IR in the Cerebral Cortex of the Head Irradiated Rats / 체질인류학회지

The purpose of this study, the effects on the cerebral cortex of the rats after brain irradiation was to investigate the change of distribution and morphology of neuropeptide-Y (NPY) neurons. Radiation was produced by the linear accelerator 6MV X-ray. The animals were categorized into control and experimental groups and we use 45 Sprague-Dawley rats weighing about 200 ~250 gm. The head areas of the animals were positioned within the radiation field of 15 cmx20 cm and with the radiation depth of 2 cm. Sodium chloral hydrate-anesthetized rats were exposed to the radiation with the dose rate of 240 cGy/min. The total dose was 1,800 cGy(rad). Animals were sacrificed on 2 hours, 5 hours, 1 day, 2 days, 3 days, 1 week, weeks, 3 weeks after brain irradiation. Using ABC immunohistochemistry, morphology and distribution of neuropeptide-Y immunoreactive neurons (NPY-IR)were studied on the cerebral cortex of the control and brain-irradiated rats. We used light and confocal laser scanning microscopy (CLSM). The following results were obtained : 1. On control group, NPY-IR neurons were found in all layers of the primary motor and sensory cerebral cortex, and the NPY-IR neurons were concentrated within the layer II, III, IV, V and VI. The typical NPY-IR perikarya was bipolar and multipolar shape. 2. On 2 hours, 5 hours after X-irradiation, decreased number of NPY-IR neurons were detected in the primary motor and sensory cerebral cortex of the rats. Also shrunken and transformed NPY-IR neurons were detected in the primary motor and sensory cerebral cortex of the rats. 3. On 1 day, 2 days, 3 days after X-irradiation, morphology and distribution of NPY-IR neurons in the primary motor and sensory cerebral cortex was generally restored. 4. On 1 week, 2 weeks, 3 weeks after X-irradiation, morphology and distribution of NPY-IR neurons in the primary motor and sensory cerebral cortex was almost similar to control group. 5. In optical serial section analysis of NPY-IR neurons, high intensity of immunofluorescence were observed in a part of the 8 ~11 sections of the control and all irradiated groups. In optical single section analysis of NPY-IR neurons, red color (high fluorescence intensity) were observed in a part of 6, 7 sections of the control and all irradiated groups. From the above results, it was concluded that the release of neurotransmitters and transcapillary leakage of blood substance were occurred on 2 hours, 5 hours, 1 day after X-irradiation, but the condition was generally restored on 3 days and 7 days following X-irradiation.

The Effect of X-irradiation on the Developing Incisor Tooth of the Rat / 체질인류학회지

In the development of tooth, the first sign is the localized thickening of oral ectoderm to form the tooth bud, and then through the dialogue between the bud and underlying mesenchyme, proliferation and differentiation of bud cells and surrounding mesenchymal cells continue, and cap and bell stages follow. In each step of development, various teratogens may act directly and indirectly, and may result a certain congenital anomalies. To reveal the action mechanism of ionizing radiation on odontogenesis morphologically, 4 Gy X-ray irradiated on the rat (Sprague-Dawley strain) fetus on GD 12.7, and observed the histological changes of the upper incisor tooth from GD 13.5 to GD 20.5, daily. In the normal development of upper incisor tooth of rat, the bud stage was from GD 12.5 ~GD 15.5, the cap stage was from GD 16.5 to GD 17.5, and the bell stage was GD 18.5 to GD 20.5. After X-irradiation on GD 12.7, the development of incisor tooth was delayed markedly, the bud stage was prolonged from GD 13.5 to GD 17.5, and the cap stage was GD 18.5 and the bell stage was from GD 19.5 to GD 20.5. After X-irradiation, from GD 13.5 to GD 16.5, apoptosis is observed in the dental organ and surrounding mesenchyme, hemorrhagic necrosis began from GD 16.5 at the center of dental pulp, followed ischemic necrosis of dental organ and surrounding mesenchyme. After GD 19.5, the development of tooth was ceased. These suggest that at least 2 mechanisms involved during X-ray teratogenesis on developing tooth. After X-irradiation on GD 12.7, initially, X-ray induced apoptosis of the cells of dental lamina and dental bud, which resulted the delayed proliferation and differentiation of dental bud and shortage of the number of cells having signal molecules which induce aggregation of the underlying mesenchymal cells. Lately, disorganization of the endothelial cells differentiated from the damaged mesenchymal cells, which resulted in rupture of capillaries and the hemorrhagic necrosis.

The Effect of X -irradiation on Lens Development / 체질인류학회지

For the study the effects of X -ray irradiation on the development of lens and interactions between optic vesicle and lens, 200 rads X -ray irradiated to the preganat rats (Sprague -Dawley strain) on gestation day 9.5. The rats were sacrificed daily from the gestation day 10.5 to 17.5, and investigated the development of lens with light microscope. The results were as follows; 1. The survival rate of X -irradiated rat fetuses was decreased significantly compared with that of the control group. 2. The mean body weight of the X -irradiated rat fetuses was decreased significantly compared with that of the control group. 3. The congenital anomalies of the eye in the rat fetuses after X -irradiation were anophthalmia, congenital aphakia, lens aplasia, lens luxation, and coloboma of the retina. 4. The critical period of lens development of the Sprague -Dawley rats was considered from GD 9. 5 to GD 10.0. 5. In the X -irradiated rat fetuses on gestation day 9.5, optic vesicle did not developed, but lens developed independently , and moved deeply toward the forebrain vesicle. According to above results, it could be concluded that the development of lens from surface ectoderm might be independent from the inductive influence of optic vesicle, and the movement of lens toward the forebrain vesicle could be mediated by chemotactic factors secreted from the brain vesicle. Further studies are needed to reveal the mechanisms of lens development and interaction between lens and brain vesicle.

Anti-tumor Effect of Combined Betacarotene with X-irradiation in the Mouse Fibrosarcoma : Cytotoxicity and Tumor Growth Delay / 대한방사선종양학회지

PURPOSE: To investigate whether combined beta-carotene with X-irradiation has more enhanced radition response than X-irradiation or not, we performed a experiment about in vitro cytotoxicity of beta-carotene and/or X-irradiation in the fibrosarcoma cells, tumor growth delay of combined beta-caroten with/or X-irradiation in the mouse fibrosarcoma. MATERIALS AND METHODS: 2% emulsion of beta-carotene was serially diluted and used. X-irradiation was given by 6 MeV linear accelerator. The cytotoxicity of beta-carotene in vitro was evaluated from clonogenic assay. To compare the cytotoxicity between combined beta-carotene with X-irradiation and X-irradiation group, 2 mg/ml of beta-carotene was contacted to fibrosarcoma (FSaII) cells for 1 hour before X-irradiation. For the tumor growth delay, single 20 Gy was given to FSaII tumor bearing C3H/N mice whic was classified as beta-crotene with X-irradiation group (n=6) and X-irradiation alone group (n=5). 0.2 ml of 20 mg/kg of beta-carotene were i.p. injected to mice 30 minute before X-irradiation in the beta-crotene with X-irradiation group. The tumor growth delay defined as the time which reach to 1,000 mm3 of tumor volume. RESULT: (1) Cytotoxicity in vitro; 1) survival fraction at beta-carotene concentration of 0.002, 0.02, 0.2 and 2 mg/ml were 0.69+/-0.07, 0.59+/-0.08, 0.08+/-0.008 and 0.02+/-0.006, respectively. 2) each survival fraction at 2, 4, 6 and 8 Gy in the 2 mg/ml of beta-carotene + X-irradiation group were 0.13+/-0.05, 0.03+/-0.005, 0.01+/-0.002 and 0.009+/-0.0008, respectively. But each survival fraction at same irradiation dose in the X- irradiation group were 0.66+/-0.05, 0.40+/-0.04, 0.11+/-0.01 and 0.03+/-0.006, respectively( p0.05). CONCLUSION: The contact of beta-caroten to FSaII cells showed mild cytotoxicity which was increased according to concentration. The cytotoxicity of combined beta-carotene with X-irradiation more increased than that of X-irradiation, additionaly. And there was significant difference of cytotoxicity between two groups. But there were no significant difference of the growth delay of fibrosarcoma between two groups.

Cytotoxic Effect of X-irradiation of Mouse Tumor Cells in the Presence of Korean Ginseng Extract / 대한방사선종양학회지

PURPOSE: We already reported the results that aqueous extract of Korean ginseng roots showed a marked cytotoxicity. In this study, we investigated whether combined ginseng product with X-irradiation increase the cytotoxicity of tumor cells than X-irradiation or not. MATERIALS AND METHODS: Fifty gram of Korean ginseng powder mixed with 1 L of distilled water was extracted with reflux flask under condition of 100 degrees C for 5 hrs. This aquaous ginseng extract was filtered, centrifuged and then was freezed under condition of -90degrees C for 16-18 hrs. The freezing extract was dried with freeze drier, and then diluted. X-irradiation was given to tumor cells by 6 MeV linear accelerator. The cytotoxicity of ginseng in vitro was evaluated from its ability to reduce the clonogenecity of fibrosarcoma (FSa II) cells. In X-irradiation alone group, each 2, 4, 6 and 8 Gy was given to tumor cells. In X-irradiation with ginseng group, 0.2 mg/mL of ginseng extract was exposed to tumor cells for 1 hour before X-irradiation. RESULTS: The yield for 50 g of ginseng extract which was treated with freezing drier was 3.13 g (6.3%). Cytotoxicity in vitro was measured as survival fraction which was judged from the curve, at ginseng concentration of 0.001, 0.01, 0.1 and 1 mg/mL were 0.89+/-0.04, 0.86+/-0.06, 0.73+/-0.01 and 0.09+/-0.02, respectively. Survival fraction at X-irradiation alone of 2, 4, 6 and 8 Gy were 0.81+/-0.07, 0.42+/-0.08, 0.15+/-0.02, 0.03+/-0.01, respectively. But, survival fraction in combined group of X-irradiation and ginseng (0.2mg/mL) at each same radiation dose were 0.28+/-0.01, 0.18+/-0.03, 0.08+/-0.02, 0.006+/-0.002, respectively ( p<0.05). CONCLUSION: The yield for ginseng extract which was treated with freezing drier was 6.3%. Cytotoxicity of Fsa II in combined ginseng with X-irradiation group was increased than that of X-irradition alone group, and its enhancing effect seemed to be added.

Radiation Effects on the Ultrastructure of Rat Cerebellar Cortex / 대한해부학회지

Severe irradiation on head may result functional alterations of central nervous system. In this study, the irradiation effect on the cerebellar cortex following heavy X-irradiation on head was studied ultrastructurally. Radiation was produced with the linear accelerator ML-4MV[Mitshubishi Co.], and rats weighing about 200gm each were exposed their heads within the radiation areas of 30cm x 30cm, under the radiation distance of 80cm, and with the radiation depth of 1.2 cm. Radiation doses were 3,000rads or 6,000rads, respectively. Animals were sacrificed on 6 hours, 2 days or 6 days following the radiation. Under anesthesia, animals were perfused with 1% glutaraldehyde-1% paraformaldehyde solution. Two hours after the perfusion, brain were taken out and refixed over night in the perfusion fixative. Small blocks of cerebellar hemispheric cortices were refixed 2 hours in 2% osmium tetroxide solution. Fixed tissues were dehydrated in alcohol, embedded in araldite mixture, and cut with ultratome. Ultrathin sections were contrasted with uranyl acetate and lead citrate solutions, and observed with electron microscope. The results obstained were as follow : 1. On 6th hour following X-irradiations, many cerebellar cortical neurons showed increased electron densities, more complicated nuclear infoldings, depletion of synaptic vesicles, expansion of astroglial territories, etc. 2. On 2nd day following X-irradiations, many organelle-rich cells such as Purkinje cells and Golgi cells were darkly degenerated. Numerous myelin figures formed by the cisternal fusions of Golgi apparatus or granular endoplasmic reticula were observed. Cytoplasmic processes of activated astroglial cells were expanded around capillaries and between granule cells. 3. On 6th day following X-irradiations, morphology of neuropil and neurones in the cerebellar cortex was generally restored, except the expanded territories of astroglial cells. From the above results, it was concluded that the release ofneurotransmitters and transcapillary leakage of blood substance were occurred on 6 hours after heavy X-irradiations. And severe alterations were produced on 2 day after X-irradiation, but the condition was generally restored on 6th day following X-irradiation.

The effect of X-irradiation on adhesion molecules and their related biological characteristics of A549 human lung adenocarcinoma cell line / 重庆医科大学学报

Objective:To investigate the effect of X-irradiation on Adhesion molecules and their related biological characteristics of Human Lung.Methods:A549 cells were irradiated with varying doses of X-rays.MTT assay was employed to evaluate the effects of X-irradiation on the proliferative inhibition of A549 cells.The immunocytochemical method was employed to analyze the protein expression levels of E-cadherin and ?-catenin.The transwell chambers were employed to evaluate the invasion of A549 cells.Results:X-irradiation could inhibit the proliferation of A549 cells in a dose-dependent manner.The protein expression of E-cadherin and ?-catenin in A549 were higher than that in control group(P