Recent studies suggest that human adipose tissue contain pluripotent cells similar to bone marrow-derived stromal cells (BSCs)^1-3. Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice⁴, we have previously demonstrated that BSCs differentiate into a variety of cell lineages both in vitro and in vivo⁵. In the present study, we extend this approach to characterize adipose-derived stromal cells (ASCs)⁶. These cells derived from human are sometimes called processed lipoaspirate (PLA) cells. ASCs were prepared from inguinal fat pads of GFP transgenic mice after extensive washing with PBS and treatment with collagenase. After the primary culture in control medium (DMEM+10%FBS), the cells were incubated in either chondrogenic medium (DMEM+1%FBS+insulin+ascorbate 2-phosphate+TGF-beta 1) or osteogenic medium (DMEM+10%FBS+dexamethasone+ascorbate-2-phosphate+beta-glycerophosphate) for two to four weeks. Chondrogenic differentiation was assessed by Alcian blue staining, while osteogenic differentiation was by von Kossa and Alkaline phosphatase staining. ASCs incubated in chondrogenic medium induced Alcian blue positive cells. Incubation with osteogenic medium became positive for von Kossa and Alkaline phosphatase staining. No osteochondrogenic differentiation was observed in cells incubated with control medium. This cell population can be easily identified through fluorescence microscope, it should be an ideal source of ASCs for further experiments of stem cell biology and tissue engineering.

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To examine the relationship between health practice in combination with health examination data and mortality, a population-based prospective cohort study was conducted in 9 towns and villages of Gunma Prefecture, Japan. Public health nurses interviewed inhabitants about their health practices. This cohort consisting of 7,694 subjects aged 40∼60 years old at baseline in 1993 was followed. During the ten-year follow-up period, 99 males and 80 females deceased. The relative risks (RRs) of some factors were estimated by the Cox proportional hazards model. RRs and 95% confidence interval with multivariate adjustment for all-cause mortality were observed significantly for age in years at the baseline (1.08, 1.04∼1.12, p<0.01), GOT (1.02, 1.00∼1.05, p<0.05), GPT (0.98, 0.96∼1.00, p<0.05), and no proteinuria (0.36, 0.14∼0.91, p<0.05) in male subjects. In females, RRs (95% confidence interval) of age in years at baseline, GOT, and γ-GTP on mortality showed significance, which were 1.09 (1.05, 1.14, p<0.01), 1.03 (1.01, 1.05, p<0.01), and 1.01 (1.00, 1.01, p<0.05), respectively. When seven health practices and age in years at baseline were used as covariates, RRs (95% confidence interval) of age and no smoking on mortality showed 1.10 (1.06, 1.14, p<0.01) and 0.57 (0.37, 0.89, p<0.05), which were significant in males. In females, significant RRs (95% confidence interval) of age, no smoking and regular exercise on mortality were observed, which were 1.11 (1.07, 1.15, p<0.01), 0.48 (0.24, 0.94, p<0.05), and 1.63 (1.01, 2.63, p<0.05), respectively. These results suggested that increased risk of death was independently associated with aging, GOT, and smoking in males and females. In addition, there was a sex difference in lifestyle-related factors such as exercise, and health examination data such as proteinuria contributing to mortality in middle-aged general inhabitants in rural towns in Japan.

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In an attempt to ascertain the effects of mechanical stimulation on callus in the early stage of bone fracture healing, a tibial fracture was induced in rats and mechanical stimulation applied to the fractures. The callus was then measured quantitatively, while the fractures were analyzed both radiographically and histologically. Following the induction of a closed transverse fracture in the tibia, external anchors were applied and the rats raised by suspending the fractured leg. The rats were divided into two main groups: a Stimulation Group (S Group) and a Control Group (C Group) without the application of any mechanical stimulation. The S Group was further divided into the following three subgroups: an axial compression group (Sc Group) receiving stimulation in the positive direction; an axial distraction group (Sd Group) receiving stimulation in the negative direction; and an axial dynamization group (Sdy Group) receiving stimulation in both directions alternately. For mechanical stimulation, 1.4-N sine waves were applied continuously for 30 minutes a day, three times a week, starting 2 days after fracture-inducing surgery. At 3, 7, and 14 days after surgery, transverse sections of each fractured bone sample were prepared. At 14 days after surgery, each transverse section was divided into two peripheral and central regions to permit calculation of the area ratio of callus. Radiographically, no marked differences were observed among the groups; histologically, differences were seen 7 days after surgery, suggesting that mechanical stimulation facilitated bone healing soon after surgery. At 14 days after surgery, the amount of callus for the C Group was less than that for all three stimulation groups. In the C Group, the amount of callus in the peripheral region was greater than in the central region, and in the Sc Group, the results were the same: callus in the peripheral region was greater than in the central region. In the Sd Group, callus was greater in the central region than in peripheral regions. In the Sdy Group, favorable callus was observed in both the central and peripheral regions. These findings suggest that axial compression facilitates callus primarily in the peripheral region, while axial distraction facilitates callus primarily in the central region. When axial compression and distraction were alternated (dynamization), callus was significantly facilitated in both the central and peripheral regions. Of the three axial stimulation techniques, dynamization was the most effective in facilitating callus in the early stage of bone fracture healing.

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Five- and ten-year survival rates in patients who had transurethral resection for pT2aN0M0 muscle invasive transitional cell carcinoma of the bladder was studied with emphasis on the effect of adjuvant transarterial infusions of anticancer agents (TAI). A total of 17 patients out of 290 patients with bladder cancer who had initial treatment in our department between January 1991 and May 2001 were found to be pT2aN0M0 and were included in the present study. Fifteen patients received intravesical instillations of BCG after transurethral resection of the bladder tumor and 8 patients received TAI which was given two weeks after bacillus Calmette-Guerin therapy, with the exception of 1 patient. Five- and ten-year overall survival rates were 76.0% and 52.1%, respectively, and both five- and ten-year cancer-specific survival rates were 76.0% with an average follow up of 71 months. Significant prognostic factor associated with survival rates was only the infiltration pattern of the tumor cells between the alpha and beta group and gamma group (P=0.0420) in cancer-specific survival rates. The data support conservative management of transurethral resection followed by intravesical instillation of BCG and TAI in patients with pT2a bladder cancer.

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To clarify the role of the genetic mutations in the clinical course of acute myeloid leukemias (AML), we analyzed for p51, p53, FLT3 and N-ras gene mutations and the expression of the p51 gene in relapsed AML. Paired samples obtained from patients with AML at both stages of diagnosis and first relapse were analyzed. Twenty-four patients with relapsed AML survived for 6 to 81 months (median 24 months) from diagnosis. In one patient, no point mutation of the p51 gene was detected, but loss of the p51 gene expression was observed at both stages. Point mutations of the p53 gene were positive at both stages (+/+) in two patients and negative at diagnosis and positive at relapse (-/+) in two patients. Tandem duplication of the FLT3 gene was detected in five patients at both stages (+/+). N-ras gene mutations at both stages (+/+) were detected in three patients. Mutant p53 at relapse was associated with short survival in patients with relapsed AML (P<0.014). Our findings show that p53 mutations were, at least in part, associated with the mechanism of relapse in AML, while new p51, FLT3 and N-ras gene alterations did not occur at relapse. Loss of the p51 gene expression in de novo AML has not been reported yet.

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Background: The major complication of Kawasaki coronary disease is myocardial infarction caused by thrombus formation inside the aneurysm or by organic obstructive lesion following the regression of aneurysm, while the indications for surgical therapy remain controversial. We have adopted coronary artery bypass grafting (CABG) even in young children for giant coronary aneurysms (more than 8 mm diameter) with or without a stenotic region when myocardial ischemia is detected. We hypothesized that a shorter time-period from diagnosis of acute Kawasaki disease (KD) to CABG would lead to better postoperative results. To elucidate the validity of our strategy, we evaluated preoperative patient characteristics and long-term outcome. Methods: Twenty-one patients (mean age: 12.0 years old) with Kawasaki coronary disease had undergone CABG during the last 12 years. The mean age at the time of acute KD was 2.7 years and the mean time range from diagnosis of acute KD to CABG was 8.1 years. The incidence of preoperative reduced ventricular function was 10 per 21 patients (47.6%). A multivariate logistic regression analysis using patient characteristics showed that the time range from acute KD to CABG was the only predictor for ventricular functional deterioration (p=0.03, odds ratio 1.55. 95%CI: 1.033∼2.325). Based on these results, we divided the patients into two groups of short time range (mean: 3.7 years; group S) and long time range (mean: 13.9 years; group L). Results: Preoperative left ventricular functional deterioration was recognized more frequently in group L (9/9, 100%) than in group S (1/12, 8.3%)(p<0.01). Myocardial infarction was documented significantly higher in the group L (6/9, 66.7%) than group S (1/12, 8.3%)(p=0.04). There was no surgical mortality in either group. The arterial grafts demonstrated good potential for growth and graft patency was 96.9%. Moreover, seven of the giant aneurysms proximal to the graft anastomosis showed complete thrombotic occlusion after CABG without development of myocardial infarction. The cardiac events free rate of group L and group S was 66.7% and 100%, respectively, during the postoperative follow up periods of 5.5±1.1 years (group L) and 4.7±1.1 years (group S). Conclusions: We successfully applied CABG for Kawasaki coronary disease. Based on our experience, a short interval after acute KD appears to be ideal for surgical treatment of Kawasaki coronary disease.

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A 42-year-old man presented with dyspnea and common cold-like symptoms. His electrocardiogram showed complete atrioventricular block and chest radiography revealed cardiomegaly and pulmonary congestion. He had a history of three recurrences of active myocarditis proven by endomyocardial biopsy over a 14-year period. Viral studies showed no evidence of recent infection, and no medications had been taken prior to this episode. This myocarditis was suggested as an unknown etiology.

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We present two Japanese students with thalassemia identified during screening for anemia in their junior high school. Blood test results revealed marked hypochromic and microcytic erythrocytosis in one patient and microcytic anemia in the other. Both cases showed a mean corpuscular volume/red blood cell (MCV/RBC) ratio less than 13. Their β/α synthesis ratio was elevated. Deletion of ψα2, ψα1, α2, α1 and &thetas;1 genes in the α-globin gene clusters were noted in the first case. This pattern of gene deletion was consistent with heterozygous α-thalassemia 1 of the Southeast Asian type. On the other hand, an increased hemoglobin A₂ level and reduced β/α synthesis ratio were found in the second case. Direct cloning and DNA sequencing identified a point mutation (guanine to adenine) at position 1 of intervening sequence II in the β-globin gene (IVS II-1 G→A). These results suggest that this patient had heterozygous β⁰-thalassemia. Diagnosis of thalassemia should be confirmed by molecular analysis in cases with microcytic anemia or hypochromic microcytosis with a MCV/RBC ratio of 13 or less.

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This is the first case report of malignant fibrous histiocytoma of the hypopharynx in a young adult with preservation of the larynx. Although the initial histopathological diagnosis of the biopsy was benign fibrous histiocytoma, subsequent histopathological evaluation of the surgical specimen was malignant fibrous histiocytoma. Thereafter, re-excision of the histiocytoma with a wide margin was performed coupled with reconstruction of the hypopharynx while preserving the larynx. We also report the good long-term survival of this case of low-grade type of malignant fibrous histiocytoma.

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