Ischemia-reperfusion (I-R) injury continues to be a problem for successful free tissue transfer and replantation after prolonged periods of ischemia. It has been shown that leukocytes and vascular endothelial cells release a variety of inflammatory mediators during reperfusion after ischemia. Leukocyte endothelial adhesion molecule-1 (LECAM-1) is a ligand on the endothelium for some adhesion receptors on leukocytes. The purpose of this study was to evaluate the blockage of leukocyte-endothelial adhesion by a monoclonal antibody (MAb) to LECAM-1 in skin flaps to prevent I-R injury in rats. Male SD rats (225-250 gr) were used. A skin flap (45×30 mm) supplied by the superficial epigastric A&V including the femoral vessels was isolated unilaterally. The femoral vessels were cross-clamped with the epigastric vessels for 9 hours of ischemia. Animals in the treated group received MAb to LECAM -1 i.v. 15 minutes prior to reperfusion; those in the control group received normal saline. Skin flap viability was assessed by tracing the outline of viable and nonviable areas. Data were collected for the following 7 days. These data corroborated with histological evidence on comparable areas of the flap. Tracing analysis revealed an average flap survival area of 89.9+/-24.7% in the treated group and 18.3+/-19.6% in the control group (p<0.005) . Histopathologically, few inflammatory changes could be observed in the treated group, while marked damage was observed in the control group. From this study, we conclud that treating skin flaps with Mab to LECAM -1 was effective for I-R injury after 9 hours of warm ischemia.

View full abstract

With an increase of breast-conserving surgery, neoadjuvant chemotherapy has become more important although there is a suitable chemotherapy regimen. The aim of this study is to evaluate the mRNA expressions of thymidine synthase (TS), dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP) and orotate phosphoribosyltransferase (OPRT) in breast carcinoma and to determine the correlation of the mRNA expressions with clinicopathologic factors. In this study, TS, DPD, TP and OPRT gene expressions were measured in 223 primary breast carcinomas between 1995 and 1999, using the Danenberg tumor profile. Laser-captured microdissection of malignant tissues was performed in formalin-fixed paraffin-embedded specimens. After extraction of RNA, the gene expressions of TS, DPD, TP and OPRT were measured by the real-time reverse transcriptional PCR method. The DPD gene expression was significantly lower in the postmenopausal tumor than in the premenopausal tumor (p=0.049), and also significantly lower in the>2cm tumor than in the ≤ 2cm tumor (p=0.048) . The DPD gene expression was significantly lower in the lymph node positive tumor than in the lymph node negative tumor (p=0.010) . TS, TP and OPRT gene expressions showed no significant correlation with clinicopathologic factors, but the TP gene expression was lower in the tumor negative for estrogen receptor or progesteron receptor than in the tumor positive for these receptors. These results suggest that the DPD gene is the most important 5-FU metabolic gene in breast carcinoma, and the neoadjuvant regimen including 5-FU may be useful for postmenopausal patients with>2cm carcinoma in the breast with lymph node involvement.

View full abstract

The number of pediatric patients with food allergies has recently been on the rise. One reason for the increase is the fact that the pathogenetic mechanisms of allergies are still poorly understood since any animal models to elucidate the pathology have not been established. In this study, therefore, we administered multiple doses of food allergens to mice in an attempt to establish a mouse model for food allergy in addition, we compared the new model with an existing model with respect to histopathological changes in the pathology. We used food allergy model mice sensitized with ovalbumin (OVA) to histopathologically examine the small intestine of mice in the control group, in the 5-week OVA short-term administration group, and in the 11-week OVA long-term administration group. The assessment items were as follows: length of villi; number of intraepithelial lymphocytes (IELs), goblet cells; and number of cryptopatches (CPs) . IELs were significantly (p < 0.05) more numerous in the OVA short-term administration group than in the control group. However, no significant difference was found in IELs between the OVA long-term administration group and the control group. The number of goblet cells was significantly (p < 0.05) greater in the OVA long-term administration group than in the control group, while no significant difference was found in the number between the OVA short-term administration group and the control group. The increased number of goblet cells in the OVA long-term administration group was conjectured attributable to the mechanism of repair and defense triggered by repeated immune responses due to sensitization with OVA. We consider that further research on the relevance of long-term exposure-elicited food allergy experimental model, along with the evaluation of immune response, is required.

View full abstract

Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor of the digestive tract. In addition, high-grade GISTs are known to show wide variability in their clinical malignant behavior. Initial tumor size and mitotic index (MI) are the most useful parameters to predict malignant potential and are used in risk assessment classification to assess prognosis of primary GISTs. In this study, we examined 39 cases of GIST, which were immunohistochemically positive for CD117 (c-kit), from 58 cases of mesenchymal tumors of the gastrointestinal tract. We focused on the correlation between the risk assessment classification and other clinicopathological features to establish helpful and reproducible parameters to indicate malignant potential and to serve practically and objectively in the routine histopathological diagnosis of GIST. The results showed that there were statistically significant differences in the SMA positive groups between high and low grade GISTs. Furthermore, high mitotic index groups showed a statistically significant difference between“metastasis/ recurrence”and “non-metastasis/recurrence”groups. Results show that MI is useful to assess the malignant potential of GIST. We analyzed the internal tandem duplication (ITD) in the FMS-like tyrosine kinase-3 gene (FLT3/ ITD mutation) in GIST. However, we did not detect an FLT3/ITD mutation in any of the 28 GISTs suggesting that the FLT3/ITD mutation is not present or very rare in GIST.

View full abstract

We conducted a follow-up study in patients with DDH in whom triradiate cartilages closed after bones previously treated at our department reached adult bone age. Examinations were performed after the patients were divided into 2 groups, one group involving patients with Conservative Therapy and the other group receiving Invasive Therapy. The conservative group included 62 joints in 56 patients (males: 7, females: 49), with a mean age of 19.2 years, and involving of 3 kinds of therapy According to the Severin classification at the final examination, 43 patients belonged to Group I and the remaining 19 patients with some residual subluxation, Group II. Of these, 6 patients underwent manual reduction under general anesthesia, 3 of whom developed inverted limbus. Invasive therapy included 17 joints in 17 patients (males: 2, females: 15) . Initial operations involved 13 patients with Ludloff s method and 4 patients undergoing femoral varus-derotation osteotomy. As for 5 patients with Ludloffs method, the remaining residual subluxation led to additional surgery for correction in adolescence. The CE angles showed an improved course in patients without femoral head deformities such as coxa magna. The results of this follow-up study demonstrated that most patients undergoing reduction by RB without apparent intervening substances yielded almost normal growth even with a slightly thickening of the floor of the acetabulum, as indicated on a plain radiograph, at the time of closed triradiate cartilage following adult bone age. In contrast, the cases with onset of some residual subluxation were often observed in patients in whom reduction was inhibited by failed acquisition of the afferent femoral head due to an inverted limbus or a thickening of the limbus, resulting in manual reduction under general anesthesia or open reduction.

View full abstract

An examination of lipid metabolism disorder and human leukocyte antigen (HLA) to identify some causes of idiopathic osteonecrosis of the femoral head (hereinafter referred to as ION) was conducted. Seventy-seven patients with ION (a mean age of 45.7 years) who have been receiving treatment were eligible for this study. Twenty-two outpatients (a mean age of 60.2 years) and 29 patients with coxarthrosis (a mean age of 47.4 years) served as control patients. The study variables relating to the disorder of lipid metabolism were measurements of total cholesterol (T-cho) and triglycerides (TG) . Additionally, apoproteins (Al, A2, B, C2, C3, E), LDL- cholesterol, HDL- cholesterol, Lp (a), and RLP-C were determined in 46 patients who consented to the study among those with ION. HLA was determined in 30 patients who consented to the study among those with ION. The Student's t-test and x² test were used for statistical analyes. A level of less than 0.05 was considered to indicate statistically significant. The ION group showed a mean T-cho of 200.0 mg dl, as compared with 200.3 mg/ dl for the coxarthrosis group and 199.6 mg/dl for the control group, suggesting that there were no significant differences among the three groups. The ION group showed a mean TG of 170.8 mg/dl, as compared with 124.0 mg/dl for the coxarthrosis group and 124.2 mg dl for the control group. These results also suggested that there were no significant differences among the three groups regarding T-cho, but the ION group alone was higher than the diagnostic criteria (150 mg/dl) . Furthermore, examinations of apoprotein and lipoprotein were performed in two ION subgroups, a group treated with Steroid therapy and a group receiving Nonsteroid therapy. There were no significant differences between the two subgroups. As for HLA, 6 antigens were expressed in A Locus, 15 antigens in B Locus, 5 antigens in C Locus, 10 antigens in DR Locus, and 3 antigens in DQ antigen. Of these antigens, B35 and DQ3 antigens were frequently and significantly observed. Additional examinations were conducted after the ION group was divided into 2 subgroups, a group involving patients with Bilateral ION and a group involving those with Unilateral ION. DR9 and DQ3 antigens were frequently and significantly observed in the Bilateral group. The results of this study suggest that some disorder of lipid metabolism is present in the ION group, but it is unlikely to be associated with the onset of osteonecrosis of the femoral head. The HLA measurements showed that significant differences are observed in the DR antigen. However, the DR antigen, a molecule expressed on the surface of antigen-presenting cells, forms an antigen presentation to T cells by binding to antigen peptides, resulting in immune response induction. This finding suggests that the theory of T cellrestrictive antigens is most likely one of the causes of the onset of ION.

View full abstract

Spinal fluid was collected from 26 patients with degenerative disc diseases (11 with lumbar disc herniation and 15 with lumbar spinal canal stenosis) and 4 healthy subjects when myelography or spinal anesthesia for surgery was performed, and the levels of major cytokines that are known to cause disc degeneration were measured. Cell cytometry and fractionation were performed. Levels of IL-1β and TNF-a, TGF-β were analyzed by ELISA: matrix metalloprotease (MMP) -3 and MMP-9, a tissue inhibitor of metalloproteinase (TIMP) -1 and TIMP-2 by enzyme immunoassay; and IL-6 by chemiluminescent enzyme immunoassay. Disc degeneration was evaluated by MRI and classified into 5 grades (from 0 to 4) . In the clinical evaluations prior to and post-surgery, the Japanese Orthopaedic Association scoring system was used. The levels of MMPs in the spinal fluid from the patients with lumbar disc herniation and lumbar spinal canal stenosis were significantly higher than those in the spinal fluid from healthy subjects. The severity of disc degeneration was not associated with the levels of MMPs. Of the patients with lumbar spinal canal stenosis, levels of MMPs were higher in patients with cauda equina syndrome than in patients with radiculopathy. The levels of IL-6 and TIMP-1 were significantly higher in patients with lumbar spinal canal stenosis (in both patients with cauda equina syndrome and those with radiculopathy) than in those with lumbar disc herniation. There was no significant difference between patients with lumbar spinal canal stenosis and those with lumbar disc herniation with respect to the number of cells and cell fractions. These results indicate that MMPs, TIMPs and the above-mentioned cytokines are also produced in the spinal canal and are involved in the degeneration of discs and spinal nerves.

View full abstract

We measured serum Diacron-reactive oxygen metabolites (D-ROM: metabolites of hydroperoxide) levels to assess the quantity of oxidative stress in climbers. We performed two independent studies. (A) Blood was collected from 14 healthy subjects before and after climbing Mt. Kita and serum D-ROM levels were measured. Results show that serum levels of D-ROM in healthy climbers after climbing were significantly higher than those before climbing. (B) Subsequently, serum levels of D-ROM in patients suffering from AMS were measured. Subjects were 21 patients who climbed Mt. Kita and consulted our mountain clinic. Serum levels of D-ROM in AMS patients were higher than those in healthy controls in Tokyo. Serum levels of D-ROM in AMS patients tended to correlate with their AMS scores. There is hypoxic and hypobaric air in the highlands which makes the endothelium produce reactive oxygen species and several inflammatory mediators known to be related to the pathogenesis of AMS. Our study suggested that a few samples from mountain climbers to monitor serum levels of D-ROM as indications of oxidative stress might serve as an assessment of the prospect and level of severity of AMS.

View full abstract