Oral administration of antigen has long been used in the induction of immune tolerance in various animal models of autoimmune diseases including rheumatoid arthritis (RA). Alleveation of arthritogenic symptoms has been reported from RA patients who received oral administration of type II collagen (CII) without side effects, however its rather inconsistent therapeutic efficacy and variation among patients calls for more detailed investigation on the mechanism of oral tolerance to be settled as regular treatment for RA. In an attempt to understand the immunogenic processes underpinning tolerance induction by orally administered CII, we analyzed changes in the expression of costimulatory molecules and STAT/SOCS signaling messengers in the mouse model of collagen induced arthritis (CIA). We found thatin the spleen of CIA mice, that has been undergone repeated oral feeding of CII prior to the induction of arthritis, showed increased promortion of CTLA4 expressing lymphocytes than in the spleen of PBS fed control. On the other hand, cells expressing CD28 or ICOS were decreased in the spleen of tolerized mice. Tolerance induction by oral CII administration also enhanced the expression of STAT6 in both RNA and protein level, while not affecting the expression of STAT3. The expression of SOCS3, which hasbeen known to transmit STAT-mediated signals from Th2 type cytokines, remained unchanged in the spleen of tolerized mice. Interestingly transcript of SOCS1, which has been associated with Th1 related pathways, was only visible in the spleen of tolerized but not of control mice, suggesting that as in the case of IL-6 signaling, it may exert a feed back inhibition toward the Th1 type stimulation.
Administration, Oral* ; Animals ; Arthritis* ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Collagen ; Collagen Type II* ; Cytokines ; Hand ; Humans ; Immune Tolerance ; Interleukin-6 ; Lymphocytes ; Mice* ; Models, Animal ; RNA ; Spleen

Background: In our study, a silver diamine fluoride (SDF) compound for the treatment of dental caries was synthesized to characterize its remineralization activity upon direct application to deciduous teeth. This study aimed to use microscopic computed tomography (microCT) and nanoindentation to evaluate whether SDF composite application could effectively arrest dental caries in five exfoliated primary molars. Methods: Carious teeth were extracted and visually examined using quantitative photofluorescence devices (Qraycam and QraypenTM). After performing microCT, the SDF composite was applied to the teeth according to the manufacturer’s instructions. The researchers exchanged and precipitated the irritant saliva once daily for 1 week. The teeth were sectioned longitudinally through the centers of the mesial and distal surfaces, embedded, polished, and measured using nanoindentation. Thereafter, microCT was repeated. Statistical analyses were performed using GraphPad Prism software. Results: Following SDF composite application, a remineralized layer was observed on microCT images, and the hardness increased when measured using nanoindentation. We found that demineralized enamel presented with an increased number of irregular crystals in the deep carious lesion group compared with those in the shallow carious lesion group, resulting in a rougher surface. Conclusion The SDF composite may be used for remineralization of early caries and cessation of advanced caries in primary molars.

OBJECTIVE: To quantify the soluble Fas ligand (sFasL) and to measure FasL-Fas complex and FasL-IgG complex in the sera of patients with various rheumatic diseases: systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), systemic sclerosis (SSc), and adult onset Still? disease (AOSD). METHODS: Serum samples were obtained from 37 patients with SLE, 40 with RA, 30 with SSc, 20 with AOSD, and 40 healthy controls. The serum sFasL, FasL-Fas complex, and FasL-IgG complex were measured using a sandwich enzyme-linked immunoabsorbent assay. Hospital medical records were retrospectively reviewed for clinical and laboratory characteristics in patients with SLE. Disease activity in SLE patients was assessed by the SLE Disease Activity Index (SLEDAI) score. RESULTS: In patients with SLE, serum sFasL levels (383.1+/-208.9pg/ml) were significantly higher (p<0.001) than those of healthy controls (192.0+/-84.7pg/ml). sFasL levels in patients with RA (150.8+/-30.7pg/ml, p=0.014), SSc (115.4+/-13.5pg/ml, p<0.001), and AOSD (137.5+/-12.9pg/ml, p=0.001) were significantly lower compared with healthy controls. The frequencies of positive FasL-Fas complex and FasL-IgG complex were higher in patients with SLE (56.8%, 56.8% respectively) than in healthy controls (2.5%, 0% respectively) (p<0.001). All patients with RA or AOSD were negative for FasL-Fas complex and FasL-IgG complex. No patients with SSc were positive for FasL-Fas complex. On the other hand, the positive frequency of FasL-IgG complex was greater in patients with SSc (16.7%) than in healthy controls (0%)(p=0.012). Serum levels of FasL-IgG complexes in active SLE patients (OD 0.467+/-0.050) were tended to be lower than those in inactive SLE patients (OD 0.509+/-0.055)(p=0.060). SLEDAI score was tended to be negatively correlated with the serum levels of FasL-IgG complex in patients with SLE (r=-0.308, p=0.068). CONCLUSION: These results suggest that FasL may possibly play a role in the pathogenesis of SLE.
Adult ; Arthritis, Rheumatoid ; Fas Ligand Protein* ; Hand ; Humans ; Lupus Erythematosus, Systemic ; Medical Records ; Retrospective Studies ; Rheumatic Diseases* ; Scleroderma, Systemic

OBJECTIVE: To quantify the soluble Fas ligand (sFasL) and to measure FasL-Fas complex and FasL-IgG complex in the sera of patients with various rheumatic diseases: systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), systemic sclerosis (SSc), and adult onset Still? disease (AOSD). METHODS: Serum samples were obtained from 37 patients with SLE, 40 with RA, 30 with SSc, 20 with AOSD, and 40 healthy controls. The serum sFasL, FasL-Fas complex, and FasL-IgG complex were measured using a sandwich enzyme-linked immunoabsorbent assay. Hospital medical records were retrospectively reviewed for clinical and laboratory characteristics in patients with SLE. Disease activity in SLE patients was assessed by the SLE Disease Activity Index (SLEDAI) score. RESULTS: In patients with SLE, serum sFasL levels (383.1+/-208.9pg/ml) were significantly higher (p<0.001) than those of healthy controls (192.0+/-84.7pg/ml). sFasL levels in patients with RA (150.8+/-30.7pg/ml, p=0.014), SSc (115.4+/-13.5pg/ml, p<0.001), and AOSD (137.5+/-12.9pg/ml, p=0.001) were significantly lower compared with healthy controls. The frequencies of positive FasL-Fas complex and FasL-IgG complex were higher in patients with SLE (56.8%, 56.8% respectively) than in healthy controls (2.5%, 0% respectively) (p<0.001). All patients with RA or AOSD were negative for FasL-Fas complex and FasL-IgG complex. No patients with SSc were positive for FasL-Fas complex. On the other hand, the positive frequency of FasL-IgG complex was greater in patients with SSc (16.7%) than in healthy controls (0%)(p=0.012). Serum levels of FasL-IgG complexes in active SLE patients (OD 0.467+/-0.050) were tended to be lower than those in inactive SLE patients (OD 0.509+/-0.055)(p=0.060). SLEDAI score was tended to be negatively correlated with the serum levels of FasL-IgG complex in patients with SLE (r=-0.308, p=0.068). CONCLUSION: These results suggest that FasL may possibly play a role in the pathogenesis of SLE.
Adult ; Arthritis, Rheumatoid ; Fas Ligand Protein* ; Hand ; Humans ; Lupus Erythematosus, Systemic ; Medical Records ; Retrospective Studies ; Rheumatic Diseases* ; Scleroderma, Systemic

BACKGROUND: Although engraftment following murine allogeneic bone marrow transplantation (BMT) is most commonly confirmed by H2 typing using flow cytometry, recipient mice can be seriously injured during peripheral blood (PB) sampling. Therefore, we developed an alternative DNA-based assay that does not require the large volume of PB necessary for flow cytometry. METHODS: A minute volume of PB from the tail vein was used to evaluate the engraftment by PCR amplification of a microsatellite in the class II Eb gene. Dilution experiments were performed to evaluate the sensitivity of this assay for detecting donor cells in mixed cell populations compared with flow cytometry analysis. RESULTS: Early engraftment and mixed chimerism were confirmed, based on the length variation of the microsatellite in the class II Eb gene. The degree of donor chimerism in the donor-recipient cell mixture could be estimated semiquantitatively in a dilution experiment. The sensitivity of this assay by the naked eye approached 10% of the degree of donor chimerism. CONCLUSION: PCR amplification of a microsatellite in the class II Eb gene can be a useful alternative to flow cytometry for evaluating early engraftment and mixed chimerism following murine nonmyeloablative BMT.
Animals ; Bone Marrow Transplantation* ; Bone Marrow* ; Chimerism* ; Flow Cytometry ; Humans ; Mice ; Microsatellite Repeats* ; Polymerase Chain Reaction* ; Tissue Donors ; Veins

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

OBJECTIVE: Hydroxychloroquine (HCQ) is a drug that has been used to treat autoimmune disorders such as rheumatoid arthritis and systemic lupus erythematosus. However, the specific mechanism for its pharmacologic action has been largely unknown. It has been reported that dysregulation of lymphocytic apoptosis mediated by Fas ligand (FasL) and Fas is associated with the development of autoimmune diseases and HCQ induces apoptosis in peripheral blood lymphocytes. These reports suggest that HCQ may exert its pharmacologic effects through the modulation of FasL and Fas. Therefore, we are intended to investigate the effects of HCQ on the regulation of FasL and Fas. Jurkat cells or peripheral blood mononuclear cells (PBMNC) were treated with varying concentrations of HCQ. Semiquantative reverse transcription- polymerase chain reaction, Western blotting, flow cytometry, and ELISA were used for this study. HCQ at nontoxic concentrations( 50~150 micrometer) caused a dose dependent increase of FasL mRNA expression and FasL in cell lysates. HCQ inhibited the release of intracellular 40 kDa FasL by Jurkat cells which were pulse-stimulated with PHA (50 microgram/ml). Jurkat cells activated with PHA increased membrane bound FasL (mFasL) expression (24.5+/-4.3%), however Jurkat cells pretreated with HCQ(150 micrometer) followed by PHA administration did not further increase mFasL expression (26.8+/-1.6%). Addition of different concentrations of HCQ to the cultured PBMNC stimulated with PHA for 24 hours showed increase of soluble FasL (sFasL). The levels of sFasL treated with HCQ zero, 50, 150 and 300 micrometer for 24 hours were 38.6+/-3.0, 43.4+/-5.1, 77.0+/-3.6(P<0.05) and 72.3+/-8.1pg/ml(P<0.05) respectively. However, fas metabolism was not affected by HCQ. CONCLUSION: These results suggest that HCQ may exhibit its pharmacological effects by upregulation of FasL gene expression and increased production sFasL without any influence on the Fas metabolism of T cells.
Apoptosis ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Fas Ligand Protein* ; Flow Cytometry ; Gene Expression ; Humans ; Hydroxychloroquine* ; Jurkat Cells ; Lupus Erythematosus, Systemic ; Lymphocytes ; Membranes ; Metabolism* ; Polymerase Chain Reaction ; RNA, Messenger ; T-Lymphocytes* ; Up-Regulation

Purpose: This study aimed to investigate and analyze the current fertility-related practices for breast cancer patients; the results are intended to help improve the quality of life of young patients and survivors. Methods: This study collected voluntary responses to a questionnaire that was used to survey Korean breast cancer specialists. The questionnaire consisted of five categories: knowledge, practice behaviors regarding fertility preservation, barriers to discussing fertility preservation, attitude toward fertility issues, and demographics and medical background. Results: A total of 120 copies of the questionnaire were distributed; the response rate was 89%. The section of the questionnaire regarding knowledge indicated that most respondents had adequate fertility preservation knowledge for cancer patients. However, 13.1% of the respondents indicated that they thought pregnancy increased the cancer recurrence risk. Respondents’ knowledge and attitudes about fertility preservation were not correlated with actual practice. The absence of patient’s expressions (24.30%), high recurrence risk (27.10%), insufficient time in the clinic (21.50%), and hospital conditions such as no reproductive specialists or infertility clinic (16.82%), were considered major barriers to discussing fertility issues. Conclusion Although more than 50% of the respondents thought that cancer treatment is more important than fertility preservation and it is complex and difficult, the Korean breast surgical oncologists were generally encouraging when discussing fertility issues with young breast cancer patients. Hence, breast clinicians should share with young patients the updated evidence regarding the feasibility and safety of pregnancy after cancer treatment and the available options so that the best decisions can be made.

Double eyelid operation is one of the most common cosmetic surgery in orientals. Among this, the non-incision technique has been usually performed in the patient with thin upper eyelid, lesser amounts of orbital fat, less redundant skin of upper eyelid, and so on. A non-incision technique make a more smooth and natural double eyelid. The merit comes from short operating time, short recovery time, less morbidity and inconspicuous scarring, Therefore, when we decide double eyelid operation on a patient who has a congenital unilateral double eyelid, we correct the aesthetic deformity in appearance caused by an unnatural asymetric upper lid which is not double eyelid using a non-incision technique. This study includes 255 patients to whom underwent double eyelid operation of unilateral double eyelid by non-incision techniques from 1990 to 1995, ranged in age 19 to 55years and follow-up period ranged from 2 months to 2 years. Two methods of non-incision technique were used in this study. One is single stitch method and the other is continuous buried suture method. The former was used in 176 patients(70.2% ) who are young with thin upper lid and the latter was used in 79 patients( 29.8% ) with redundant or puffy upper lid. Most of the patients were satisfied with smooth and symmetric appearance. We concluded that non-incision technique can be a good surgery method of correcting unilateral double eyelid. And we had good results using the continuous buried suture method in the case of puffy upper eyelid.
Cicatrix ; Congenital Abnormalities ; Eyelids* ; Follow-Up Studies ; Humans ; Orbit ; Skin ; Surgery, Plastic ; Sutures

This study aimed to assess the influence of different cross-sectional area on the cyclic fatigue fracture of Ni-Ti rotary files using a fatigue tester incorporating cyclical axial movement. Six brands of Ni-Ti rotary files (ISO 30 size with .04 taper) of 10 each were tested: Alpha system (KOMET), HeroShaper (MicroMega), K3 (SybronEndo), Mtwo (VDW), NRT (Mani), and ProFile (Dentsply). A fatigue-tester (Denbotix) was designed to allow cyclic tension and compressive stress on the tip of the instrument. Each file was mounted on a torque controlled motor (Aseptico) using a 1:20 reduction contra-angle and was rotated at 300 rpm with a continuous, 6 mm axial oscillating motion inside an artificial steel canal. The canal had a 60degrees angle and a 5 mm radius of curvature. Instrument fracture was visually detected and the time until fracture was recorded by a digital stop watch. The data were analyzed statistically. Fractographic analysis of all fractured surfaces was performed to determine the fracture modes using a scanning electron microscope. Cross-sectional area at 3 mm from the tip of 3 unused Ni-Ti instruments for each group was calculated using Image-Pro Plus (Imagej 1.34n, NIH). Results showed that NRT and ProFile had significantly longer time to fracture compared to the other groups (p < .05). The cross-sectional area was not significantly associated with fatigue resistance. Fractographycally, all fractured surfaces demonstrated a combination of ductile and brittle fracture. In conclusion, there was no significant relationship between fatigue resistance and the cross-sectional area of Ni-Ti instruments under experimental conditions.
Electrons ; Fatigue ; Fractures, Stress ; Nickel ; Radius ; Steel ; Titanium ; Torque