No abstract available.
Animals ; Mice*

PURPOSE: Oral mucositis induced by radiotherapy to the head and neck area, is a common acute complication and is considered as the most severe symptom for cancer patients in the early stages of treatment. This study was proposed to establish the oral mucositis mouse model induced by a single dose of radiation for the facility of testing therapeutic candidates which can be used for the oral mucositis treatments. MATERIALS AND METHODS: Fifty-five BALB/c mice were divided into four groups: control, 16 Gy, 18 Gy, and 20 Gy. Oral mucositis was induced by a single dose of radiation to the head and neck using 6 MV x-Ray from linear accelerator. After irradiation, body weight and physical abnormalities were checked daily. Tongue tissues from all groups were taken on days 1, 2, 3, 5, 7, 9, and 14, respectively and H&E staining was conducted to examine morphological changes. RESULTS: Body weight dramatically decreased after day 5 in all irradiated mice. In the 16 Gy treatment group, body weight was recovered on day 14. The histology data showed that the thickness of the epithelial cell layer was decreased by the accumulated time after radiation treatment, up to day 9. Severe ulceration was revealed on day 9. CONCLUSION: A single dose of 16 Gy is sufficient dose to induce oral mucositis in Balb/C mice. Significant changes were observed in the Balb/C mice on days 7 and 9 after radiation. It is suggested that this mouse model might be a useful standard tool for studying oral mucositis induced by radiation.
Mice ; Animals

No abstract available.
Animals ; Mice*

No abstract available.
Animals ; Mice*

It has been thought that autoimmune diseases like systemic lupus erythematosus and rhumatoid athritis are closely associated with anti-DNA antibodies (Abs). In studies of the control for anti-DNA Ab generation, an understanding of the regulatory mechanisms by anti-idiotypic Abs that influence the production of anti- DNA Abs would be facillitated by the availability of the hybridomas producing the pairs of DNA-specific and anti-DNA's idiotope-specific monoclonal antibodies (mAbs). We have produced a series of anti-DNA mAbs and then monoclonal anti-idiotypic Ab directed against idiotypic determinant of the 3D8 mAb that has the highest affinity to dsDNA and ssDNA among the anti-DNA mABs that we had obtained. The spleen cells of the MRL-Ipr/Ipr, autoimmune prone, mice were fused with P3X63Ag8.653 myeioma cells to obtain anti-DNA Ab secreting hybridomas. Out of the fourteen clones that showed strong binding to ssDNA, four clones had cross-reactivity with dsDNA whereas none of these clones reacted with left-handed z-DNA. The binding activities of the anti-DNA mAbs to various synthetic polynucleotide sequences were different respectively. Anti-idiotypic mAbs were generated by the fusion of myeloma cells and spleen cells from the Balb/c mice immunized with 3DB-Fab. We have produced two anti-idiotypic mAbs, B7 (IgG2a/k) and 02F3 (IgM/k), which were specific to 3DB-Fab and cloned the variable region of the heavy chain from the 02F3 hybridoma.
Mice ; Animals

OBJECTIVE: To determine the effects of leukemia inhibitory factor (LIF) on embryonal development in in vitro culture. METHODS: This is designed in vitro model using eggs from mouse. The eggs from mouse were assigned 29 for control group, 53 for 20 ng/ml of LIF, 88 for 40 ng/ml of LIF, 68 for 80 ng/ml of LIF respectively for in vitro fertilization. And 26 fertilized eggs at 2 cell stage from mouse also were assigned. The mouse embryos of all groups were cultured in medium supplemented with LIF in different concentrations, whereas the eggs in control group was cultured in medium without supplement of LIF. RESULTS: At 72 hours culture of eggs from in vitro fertilization, there was a slight increas in rate of embryonal development to morula in both LIF-20 and LIF-40 as results of 64.15% and 75% respectively, while 42.65% in inferior rate of LIF-80, compare with 51.72% in control group. But the difference between these each groups were not significant in statistically (p< or =0.05). And after 96 hours culture of eggs, the rates blastocyst formation was significantly higher in both LIF-20 and LIF-40 as 56.6% and 63.63% than those in control and LIF-80 as 44.83% and 35.29% respectively. On culturing eggs from in vivo fertilization, the rates of blastocyst formation was significantly not only higher as 85% and 81.81% respectively in medium supplemented with LIF-40 and LIF-80 than 42.3% in LIF-20 but also embryonal cell viability were remakedly improved at 96 hours after culture. CONCLUSION: The LIF in low dose is embryotrophic, but LIF in high dose is embryotoxic on eggs from in vitro fertilization. Whereas on culturing eggs from in vivo fertilization, LIF is more beneficial with dose dependent in high concentration.
Mice ; Animals

OBJECTIVE: In muscle and neuronal cells, calcium channels have been classified by electrophysiological and pharmacological properties into (1) voltage-dependent Ca2+-channel(1) P/Q-type Ca2+-channel (2) N-type Ca2+-channel R(3) L-type Ca2+-channel (4) T-type Ca2+-channel (5)R-type Ca2+-channel. The present study was done in order to investigate whether there is any difference in Ca2+-channel distribution between activated and normally fertilized embryos. METHODS: The immunocytochemical method was used to identify the existence of voltage-dependent Ca2+-channels in parthenogenetically activated 2-cell embryos by ethanol and SrCl2 treatment. These 2-cell embryos were obtained by exposure to 6% ethanol for 6 min and to 10 mM SrCl2 for 2h. RESULTS: P/Q-type Ca2+ channels and L-type Ca2+-channels have been identified. Whereas, three type of Ca2+-channel P/Q-type, N-type, L-type have been identified in 2-cell embryos fertilized in vivo. CONCLUSION: Activation by ethanol was faster than those by SrCl2. However, there was difference in DAB staining of the embryos between ethanol and SrCl2 treatment (87.7% and 54.1%). Intensity of staining was also different between ethanol- and SrCl2-treated group. However, it has not been known why there was some difference in DAB staining and staining intensity in the present study.
Mice ; Animals

PURPOSE: To investigate synergistic effect of local cryoablation with systemic immunotherapy, the tumor control ability and immunologic responses of combining these two modalities was compared with that of cryoablation, surgical excision, and immunotherapy only group in a tumor re-challenge model. MATERIALS AND METHODS: Preliminary experiments were performed in two stages. The first stage consisted of 36 Balb/c mice with Renca bearing tumors imbedded in the right thigh, and was treated with interleukin-2 (IL-2) and interferon-alpha(IFN-alpha) to evaluate the efficacy of immunotherapy and to determine the adequate dosage. The second stage was performed on 10 mice, to evaluate histological changes and efficacy after cryoablation. The main experiment was performed on 48 mice, divided into 6 groups of control with tumor implantation, excision of tumor, excision combined with immunotherapy, cryoablation of tumor, cryoablation with immunotherapy and control without tumor. After treatment, tumor re-challenge was performed with Renca cell, then the growth pattern was evaluated with physical measurements, and immune response was investigated with fluorescent activated cell sorter and cytotoxicity assay. RESULTS: Preliminary studies on immunologic efficacy revealed that IL-2 and IFN-alpha have a dose dependent inhibition of tumor growth. The main experiment evaluating the efficacy of combination treatment revealed that cryoablation with immunotherapy proved to be most effective in terms of tumor recurrence and tumor growth inhibition, yet the difference was not statistically significant from monotherapy with cryoablation. However, cytotoxicity was significantly increased cryoablation with immunotherapy compared with other groups. CONCLUSIONS: Cryoablation on tumor re-challenge mice model showed advantages with immunotherapy most prominently in cytotoxicity.
Mice ; Animals

The interaction of radiation and 5-luorouracil (5-U) on mouse jejunal crypt cells was studied using the microcolony survival assay. 150mg/kg of 5-U was injected intraperitoneally 15 minutes before irradiation and 6 hours after irradiation. Jejunal crypt cells of mouse survived more when 5-U was given 15 minutes before irradiation than giving it 6 hours after irradiation. The mean lethal doses (Do) of each of irradiation alone group, 5-U injection group of 15 minutes preceding irradiation, and 5-U injection group of 6 hours post irradiation were. 135, 135, and 114 rad respectively. The dose effect factor (DEF) of each of 5-U injection groups of 15 minutes preceding irradiation and of 6 hours post irradiation were 1.13 and 1.27.
Animals ; Mice*

OBJECTIVE: Indoleamine 2, 3-dioxygenase (IDO), an immuno suppression enzyme, is one of the initial and rate-limiting enzymes involved in the catabolism of the essential amino acid tryptophan. IDO inhibits T cell proliferation, induces T cell apoptosis, and plays a fundamental role in autoimmunity and allergy. We investigated which subtype of dendritic cells (DCs) is involved in IDO expression and the generation of regulatory T cells during the induction of oral tolerance in type II collagen-induced arthritis (CIA). METHODS: Type II Collagen was fed to DBA/1J mice before immunization. Changes in DC subtypes and induction of regulatory T cell in orally tolerized CIA mice were analyzed. Whether the effect of DC subtype was modulated by the IDO expression, was determined by flow cytometry (FACs) and confocal microscopy. RESULTS: IDO expression of CD11c+ DCs was higher in orally tolerized CIA mice than in non-tolerized CIA mice. CD11b+ DCs of the CD11c +DCs, subtype was higher in the induction of in IDO expression. Our data suggest that these IDO expressing DCs of oral tolerized mice suppressed type II collagen-specific T cell proliferation and favored the differentiation of naive CD4+ T cells into regulatory T cells. Especially, CD11c+CD11b+ DCs expressed IDO, which is known to be associated with regulatory T cell induction. CONCLUSION: We observed that oral tolerance induced the increase in IDO-expressing CD11c+CD11b+ DCs, which appeared to induce regulatory T cells. IDO-expressing CD11c+CD11b+ DCs are involved in oral tolerance, which may provide a new therapeutic approach for the treatment of rheumatoid arthritis.
Mice ; Animals