Ischemic preconditioning (IP), short pre-treatment sublethal ischemia, induces a state of protection against subsequent prolonged ischemia-reperfusion. The purpose of this study was to investigate the expression of HO-1, HSP70, and iNOS proteins in the liver subjected to the courses of reperfusion after repetitive cycles of remote IP in the rat. Using thirty five week-old rats, the remote preconditioning was undertaken by vascular clamp occlusion of blood flow to one hindlimb, with 3 and 10 cycles of 5 minutes occlusion followed by 5 minutes reperfusion. The liver was removed 0, 3, 6, 24, and 72 hours of reperfusion after remote IP and assayed by immunohistochemical staining and Western blotting analyses for anti-HO-1, anti-HSP70, and anti-iNOS antibodies. The expression of HO-1 in rat liver increased at 72 hours of reperfusion groups after 3 and 10 cycles of remote IP, compared with normal control groups. The expression of HSP70 in rat liver increased at 6 hours of reperfusion groups after 3 cycles of remote IP, compared with normal control groups. The expression of HSP70 in rat liver increased at 0 hour of reperfusion groups after 10 cycles of remote IP, compared with normal control groups and decreased at 24 and 72 hours of reperfusion groups. The expression of iNOS in rat liver increased at 24 hours of reperfusion groups, but decreased at 72 hours of reperfusion groups after 3 and 10 cycles of remote IP, compared with normal control groups. In summary, these results showed that at early phase of reperfusion after remote IP, HSP70 expression was increased in rat liver. However, at 72 hrs of reperfusion after remote IP, HO-1 expression was increased and iNOS expression was decreased in rat liver.
Animals ; Blotting, Western ; Hindlimb ; Ischemia ; Ischemic Preconditioning ; Liver ; Proteins ; Rats ; Reperfusion

No Abstract Available.
Female ; Hormone Replacement Therapy* ; Humans ; Life Style* ; Osteoporosis*

In allogeneic transplantation, including the B6 anti-BALB.B settings, H60 and H4 are two representative dominant minor histocompatibility antigens that induce strong CD8 T-cell responses. With different distribution patterns, H60 expression is restricted to hematopoietic cells, whereas H4 is ubiquitously expressed. H60-specific CD8 T-cell response has been known to be dominant in most cases of B6 anti-BALB.B allo-responses, except in the case of skin transplantation. To understand the mechanism underlying the subdominance of H60 during allogeneic skin transplantation, we investigated the dynamics of the H60-specific CD8 T cells in B6 mice transplanted with allogeneic BALB.B tail skin. Unexpectedly, longitudinal bioluminescence imaging and flow cytometric analyses revealed that H60-specific CD8 T cells were not always subdominant to H4-specific cells but instead showed a brief dominance before the H4 response became predominant. H60-specific CD8 T cells could expand in the draining lymph node and migrate to the BALB.B allografts, indicating their active participation in the anti-BALB.B allo-response. Enhancing the frequencies of H60-reactive CD8 T cells prior to skin transplantation reversed the immune hierarchy between H60 and H4. Additionally, H60 became predominant when antigen presentation was limited to the direct pathway. However, when antigen presentation was restricted to the indirect pathway, the expansion of H60-specific CD8 T cells was limited, whereas H4-specific CD8 T cells expanded significantly, suggesting that the temporary immunodominance and eventual subdominance of H60 could be due to their reliance on the direct antigen presentation pathway. These results enhance our understanding of the immunodominance phenomenon following allogeneic tissue transplantation.
Animals ; Antigen Presentation ; Antigen-Presenting Cells/immunology/metabolism ; CD8-Positive T-Lymphocytes/*immunology ; Epitopes, T-Lymphocyte/*immunology ; Female ; Graft Rejection/*immunology ; Interferon-gamma ; Lymphocyte Activation/immunology ; Lymphocyte Count ; Mice ; Minor Histocompatibility Antigens/*immunology/metabolism ; *Skin Transplantation ; Transplantation, Homologous

PURPOSE: Calcium-activated potassium channels(KCa) may be involved in the transient outward current of the first phase of cardiac action potential. But it is still not clear whether cardiac myocytes express any Kca. We try to identify here the types of Kc, expressed in rat caridac myocytes. METHODS: We isolated total heart RNA from 50 rats(Spague-Dawley) and performed reverse transcription-polymerase chain reaction(RT-PCR) using specifically designed synthetic oligonucleotide primer sets. From the pure culture of cardiac myocyte, Kc, gene expression was detected by Southern blot analysis. RESULTS: RT-PCR revealed expressions of BKca(large-conductance Kca, rSlo) and S&,(small-conductance Kca, rSK1). We prepared cardiac myocytes pure culture(>9596 pure myocyte) using pure culture technique. RT-PCR and Southern blot analysis of rat cardiac myocyte showed only rSK1-specific band, but no rSlo-specific was detected. CONCLUSION: The expressions of more than one type of Kca are detected from rat heart. A sub-type of SKcrSK1, was expressed in cardiac myocyte, while the main subunit of BKca(rSlo) was found in cells other than myocytes, most likely in the smooth muscle of cardiac blood vessels.
Action Potentials ; Animals ; Blood Vessels ; Blotting, Southern ; Culture Techniques ; Gene Expression ; Heart ; Muscle Cells ; Muscle, Smooth ; Myocytes, Cardiac* ; Potassium ; Potassium Channels, Calcium-Activated* ; Rats* ; RNA

Various cytokines and chemokines play a role in carcinogenesis. However, no study has previously been undertaken to investigate comprehensively the expressions of cytokines and chemokines in hepatoma cells. In this study, we determined which cytokines and chemokines are expressed in hepatoma cells. Recently, it was reported that the expressions of several chemokines could be increased by Fas stimulus in many normal and cancer cells. Therefore, we also investigated whether chemokines expression is regulated by Fas ligation. To address this issue, we performed RNase protection assays upon 13 cytokines and 8 chemokines genes in 10 human hepatoma cell lines, comprising 8 hepatitis B virus (HBV)-associated hepatoma cell lines. Transforming growth factor-beta2 (TGF-beta2) was found to be expressed in 8 HBV-associated hepatoma cell lines, and to be potently expressed in 5 cell lines; however, the mRNA expressions of interleukin-10 (IL-10), IL-12, interferon-gamma(IFN-gamma) and tumor necrosis factor-alpha(TNF-alpha) were not detected in any cell lines examined. Among the chemokines investigated in this study, IL-8 was expressed by 8 HBV- associated hepatoma cell lines, and monocyte chemoattractant protein-1 (MCP-1) by 7 HBV-associated hepatoma cell lines. However, the mRNA expressions of macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-beta, interferon-inducible protein-10 (IP-10), RANTES, lymphotactin and I-309 were either very weak or undetectable. Fas ligation did not increase chemokines expression in hepatoma cells. Conclusively, TGF-beta2, IL-8 and MCP-1 were overexpressed in HBV-associated hepatoma cells, and the expressions of chemokines were not increased by Fas ligation in human hepatoma cells.
Antigens, CD95/physiology ; Carcinoma, Hepatocellular/*metabolism ; Chemokines/*genetics ; Cytokines/*genetics ; Gene Expression Profiling ; Human ; Liver Neoplasms/*metabolism ; RNA, Messenger/analysis ; Tumor Cells, Cultured

Diagnosis of acute appendicitis in children is sometimes difficult. The aim of this study is to validate a clinical scoring system and ultrasonography for the early diagnosis and treatment of appendicitis in childhood. This is a prospective study on 59 children admitted with abdominal pain at St. Mary's Hospital, the Catholic University of Korea from July 2002 to August 2003. We applied Madan Samuel's Pediatric Appendicitis Score (PAS) based on preoperative history, physical examination, laboratory finding and ultrasonography. This study was designed as follows: patients with score 5 or less were observed regardless of the positive ultrasonographic finding, patients with score 6 and 7 were decided according to the ultrasonogram and patients above score 8 were operated in spite of negative ultrasonographic finding. The patients were divided into two groups, appendicitis (group A) and non-appendicitis groups (group B). Group A consisted of 36 cases and Group B, 23 cases. Mean score of group A was 8.75 and group B was 6.13 (p<0.001). Comparing the diagnostic methods in acute appendicitis by surveying sensitivity, specificity, positive predictive value, negative predictive value, and accuracy, PAS gave 1.0000, 0.3043, 0.6923, 1.0000, and 0.7288, and ultrasonography gave 0.7778, 0.9130, 0.9333, 0.7241, and 0.8300 while the combined test gave 1.0000, 0.8696, 0.9231, 1.0000, and 0.9490, respectively. Negative laparotomy rate was 3 %. In conclusion, the combination of PAS and ultrasonography is a more accurate diagnostic tool than either PAS or ultrasonography.
Abdominal Pain ; Appendicitis ; Child ; Diagnosis* ; Early Diagnosis ; Humans ; Korea ; Laparotomy ; Physical Examination ; Prospective Studies ; Sensitivity and Specificity ; Ultrasonography*

As the interest in health is increasing and the population enjoyed the leisure sports is steadily increasing, the stress fracture, fracture or variant of metatarsal bone of foot has been shown frequently. The mistaken estimation about the length and rank of metatarsal bones during the osteotomy of the metatarsal bones of foot can be complicated. It is essential to have detailed knowledges about the anatomical structure of surgical region. This study aimed to investigate the metatarsal bones of foot and to develop a regression equation that can predict the length of metatarsal bones during the osteotomy. The subject of this study is fifty four feet (30M/224F). We measured the whole length and the article length of metatarsal bone. Also, we measured the whole width and the article width in the head, body, base of the metatarsal bone. The data was analyzed using SPSS win 13.0. The regression equation models of length of the metatarsal bones were developed by multiple regression analysis. The regression equation predicted first metatarsal length was second metatarsal articular length x0.770+7.780, second metatarsal length was third metatarsal length x0.976+6.050, third metatarsal length was fourth metatarsal length x1.000+0.922, fourth metatarsal length was third metatarsal length x0.917+4.167, fifth metatarsal length was fourth metatarsal length x0.901+7.972. The results of this study would be useful to clarify the characteristics of the metatarsal bone of the foot, to develop a regression equation for prediction of the length of the metatarsal bone.
Foot ; Fractures, Stress ; Head ; Leisure Activities ; Metatarsal Bones ; Osteotomy ; Sports

BACKGROUND: Physical activity and calcium nutriture with reproductive endocrine status are primary controller of bone remodelling activity. There are differences in impact of exercise on early menopausal bone ; late menopausal bone. There are possibility of different effect of calcium intake on bone mass among different life stage. The aim of this study was to elucidate whether the relation between lifestyle and bone mineral density varied with life stages. METHODS: We examined bone mineral density and took questionnaires related to lifestyle of 1,698 women aged 49~54 years old who lived in ulsan from July 1999 to Dec. 1999. We selected 731 healthy subjects without medical conditions or lifestyle factors known to affect bone metabolism. RESULTS: In 6~10 years postmenopausal women, those with calcium intake of more than 600 or 800mg /day showed significantly greater BMD. In postmenopausal women , those daily consumption of milk showed greater BMD. But it is not significantly. In premenopausal women with regular menstruation, those who took regular exercise showed significantly greater BMD than those who did not. Working hours is not related with BMD. CONCLUSIONS: Our study showed that the relation between calcium intake or physical activity and BMD differed with life stages. It was suggested that life stages should be taken into consideration to perform lifestyle modifications for the prevention and management of osteoporosis.
Bone Density* ; Calcium* ; Female ; Humans ; Life Style ; Menopause ; Menstruation ; Metabolism ; Milk ; Motor Activity* ; Osteoporosis ; Ulsan ; Surveys and Questionnaires

Akt, heat shock protein (HSP72)72, and HSP90 induced by ischemic preconditioning protect cells from the ischemic injury. The purpose of this study was to examine the alterations of the level of phospho-Akt, HSP72, and HSP90 in the rat tibialis anterior and soleus muscles after cyclic episodes of ischemic preconditioning. Sprague-Dawley rats aged 35 weeks were divided into control and ischemic preconditioning (IP) groups. The IP group was divided into 3 subgroups based on cycles of IP. Left common iliac artery was occluded 3, 6, and 10 times for 5 minutes, followed by 5 minutes reperfusion. The experimental animals were sacrificed at 0, 3, 6, 24, and 72 hours after reperfusion, and left tibialis anterior and soleus muscles were removed. The expression of phospho-Akt, HSP72, and HSP90 were examined with immunohistochemical methods and Western blot analysis. The results were as follows; 1. In the 3 and 6 times of IP groups, the expression of phospho-Akt (p-Akt) was increased at 0 and 3 hours after reperfusion, compared with control group. The expression of p-Akt in the 10 times of IP group was lower than that in 3 and 6 times of IP groups. At 72 hours after reperfusion, the expression of p-Akt showed no difference among the IP groups. The expression of p-Akt was higher in Soleus than that in Tibialis anterior. 2. The expression of HSP72 in 3 times of IP group increased at 0 and 3 hours after reperfusion, compared with 6 and 10 times of IP groups. The expression of HSP72 in the 10 times of IP group was lower than that in 3 and 6 times of IP groups. At 72 hours after reperfusion, the expression of HSP72 showed no difference among the IP groups. The expression of HSP72 was higher in Soleus than that in Tibialis anterior. 3. In the 3 and 6 times of IP groups, the expression of HSP90 increased at 0 and 3 hours after reperfusion, compared with control group. The expression of HSP90 in the 10 times of IP group was lower than that in 3 and 6 times of IP groups. At 24 hours after reperfusion, the expression of HSP90 showed no difference with increasing episode of IP. The expression level of HSP90 was higher in Soleus than that in Tibialis anterior. These findings suggest that ischemic preconditioning increases the expression of p-Akt, HSP72 and HSP90 at early phase after reperfusion in the rat tibialis anterior and soleus muscles. However, increased cycles of ischemic preconditioning may not induce the expression of them.
Animals ; Blotting, Western ; Heat-Shock Proteins ; Iliac Artery ; Ischemic Preconditioning* ; Muscles* ; Phosphorylation* ; Rats* ; Rats, Sprague-Dawley ; Reperfusion

Remote ischemic preconditioning (IP), brief tolerating cycles of ischemia and reperfusion in remote non-vital organs, can reduce ischemic injury of the heart. IP induces cardiac protection by down-regulating iNOS or up-regulating eNOS. In addition, Akt has been known to protect myocardium against ischemia-reperfusion injury. This study was undertaken to observe the expression of iNOS, eNOS, Akt and phospho-Akt (p-Akt) in the rat myocardium after IP. Thirty-five weeks-old male Sprague-Dawley rats were divided into control and IP groups. The IP group was further subdivided into 3 groups based on the number of cycles of IP. For IP, left commom iliac artery was occluded 3, 6 and 10 cycles for 5 min of ischemia alternating with 5 min of reperfusion. The rat were sacrificed at 0, 3, 6, 24 and 72 hours of IP and the heart was removed. The expression of iNOS, eNOS, Akt and p-Akt in the rat myocardium was examined by immunohistochemical staining and Western blot analysis. The expression of iNOS was increased by IP and was higher in 10IP groups than 3IP and 6IP group. The expression of eNOS was increased or decreased by IP and was showed no difference with increasing episode of IP. The expression of Akt was decreased by IP at 24 and 72 hours after reperfusion, and showed no differences with increasing episode of IP. The expression of p-Akt was increased by IP and showed no difference with increasing episode of IP. These results suggest that hind limb ischemic preconditioning provides cardiac protection through up-regulation of eNOS and phosphorylation of Akt, however excessive episodes of remote preconditioning may induce the myocardial ischemic injury through overexpression of iNOS.
Animals ; Blotting, Western ; Extremities ; Heart ; Humans ; Iliac Artery ; Ischemia ; Ischemic Preconditioning ; Male ; Myocardium ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; Reperfusion Injury ; Up-Regulation