Although uncommon, acquired hemophilia A (HA) is associated with a high rate of mortality due to severe bleeding. In spite of many hypotheses regarding the cause of acquired HA, there is as yet no established theory. In this study, we investigated the possibility that mutation(s) in the F8 gene may be correlated with the development of inhibitory autoantibodies. Direct sequencing analysis was performed on all 26 exons of the F8 gene of 2 patients exhibiting acquired HA. Both patients were found to share a common point mutation (c.8899G>A) in the 3'-untranslated region (3'-UTR) of exon 26. This is the first report on the genotyping of F8 in the context of acquired HA.
Autoantibodies ; Exons ; Hemophilia A ; Hemorrhage ; Humans ; Point Mutation

Pulmonary alveolar proteinosis is a rare disease, which hallmark is a dense accumulation of PAS positive phospholipid material within alveolar sac. Pulmonary alveolar proteinosis is classified as primary form of unknown etiology and secondary form associated with other diseases. We report a case of secondary pulmonary alveolar proteinosis associated with acute erythroleukemia. A C year old male patient complained of nonproductive cough and general weakness, and presented fine inspiratory crackles at both lower lung field. Chest radiographs and high resolution CT scans showd a lobular pattern of ground-grass opacity with interlobular septal thickening in the center field of the both lungs, Bone marrow aspiration and biopsy revealed acute erythroleukemia. Open lung biopsy revealed PAS positive eosinophilic granular material filled in alveoli. He was treated with TAD chemotherapy, but died from multiorgan failure with pneumonia 22days after chemotherapy.
Biopsy ; Bone Marrow ; Cough ; Drug Therapy ; Eosinophils ; Humans ; Leukemia, Erythroblastic, Acute* ; Lung ; Male ; Pneumonia ; Pulmonary Alveolar Proteinosis* ; Radiography, Thoracic ; Rare Diseases ; Respiratory Sounds ; Tomography, X-Ray Computed

BACKGROUND: Acute leukemias co-expressing myeloid and lymphoid antigens but does not meet the criteria for biphenotypic acute leukemia (BAL) is common, however its clinical significance is not fully defined. METHODS: In this study, clinical features of 68 co-expressing (myeloid and lymphoid) acute leukemias diagnosed between January 2000 and December 2006 were studied and compared with those of a control group of patients (pure AML or ALL). RESULTS: Age, gender, initial Lactate dehydrogenase (LDH) level and cytogenetics were not different between the co-expressing group and the control group. But, the initial bone marrow blast percent was significantly higher in the co-expressing group (70% vs. 54.5%, P=0.003). Fifty five percent (16/29) of ALL and 30% (52/172) of AML patients showed myeloid and lymphoid markers concomitantly. The lymphoid antigen positive AML (Ly+AML) patients showed significantly shorter survival rates than pure AML patients (4 year survival rate, 17.6% vs. 45.6%, P=0.002). However hematopoietic stem cell transplantation (HST) abrogated the difference (4 year survival rate, 54.7% vs. 50.6%, P=0.894). In ALL patients, survival rate was not affected by myeloid antigen co-expression (4 year survival rate 26.1% vs. 20%, P=0.954). CONCLUSION: Co-expression of lymphoid markers in AML should be regarded as a poor prognostic factor and more aggressive treatment such as HST should be considered.
Bone Marrow ; Cytogenetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; L-Lactate Dehydrogenase ; Leukemia ; Leukemia, Biphenotypic, Acute ; Prognosis ; Survival Rate

It is important to identify therapeutic compounds with no adverse effects for use in the chemotherapy of patients with bone-related diseases. The aim of this study was to identify a new compound that inhibits osteoclast differentiation and bone resorption. Herein, we examined the effects of 1',2'-dihydrorotenone on osteoclast differentiation and bone resorption in vitro and in vivo. 1',2'-dihydrorotenone inhibited receptor activator of NF-kappaB ligand (RANKL)-induced osteoclast differentiation of cultured bone marrow macrophages (BMMs) in a dose-dependent manner. However, 1',2'-dihydrorotenone did not exert cytotoxic effect on BMMs. 1',2'-dihydrorotenone suppressed the expression of c-fos and NFATc1 as well as osteoclast-specific genes in BMMs treated with RANKL. Treatment with RANKL inhibited the expression of inhibitors of differentiation/DNA binding (Id)1, 2, and 3; however, in the presence of 1',2'-dihydrorotenone, RANKL did not suppress the expression of Id1, 2, and 3. Furthermore, 1',2'-dihydrorotenone inhibited bone resorption and considerably attenuated the erosion of trabecular bone induced by lipopolysaccharide treatment. Taken together, these results suggest that 1',2'-dihydrorotenone has the potential to be applied in therapies for bone-related diseases.
Bone Marrow ; Bone Resorption ; Humans ; Macrophages ; Osteoclasts ; Receptor Activator of Nuclear Factor-kappa B ; Rotenone

BACKGROUND: On performing umbilical cord blood (UCB) transplantation, faster engraftment may lead better clinical outcome. Because transplanted viable cell count in UCB is related to the engraftment, accurate evaluation of viability of CD34+cells in cryopreserved UCB has clinical implication. We examined the difference in viability of cells in cryopreserved UCB according to the duration of cryopreservation and different methods. METHODS: A total of 60 UCB samples which were cryopreserved for 1 to 4 years were used in this study. Viability of cryopreserved cells were examined with trypan blue exclusion assay, DNA contents analysis, caspase-3 activation test, intracellular esterase activity and Annexin-V/PI staining. RESULTS: After thawing the cryopreserved UCB, 89% of the total MNCs and 84% of CD34+cells were viable as identified by trypan blue exclusion assay. In the CD34+cell population, the cell death rate was found to be 47% by Annexin-V/PI staining and less than 5% by DNA contents analysis. However, cspase-3 activity failed to document apoptosis. The intracellular esterase activity test also showed a cell death rate of about 10~20% at 2, 4, and 6 hours after thawing. CONCLUSION: Viable cells in UCB should be measured by several compensatory techniques rather than a single method. Discordance among Annexin-V/PI staining versus trypan blue exclusion, DNA contents analysis, and the caspase-3 activation test or intracellular esterase activity should be clarified in order to apply these techniques for actual cord blood transplantation.
Apoptosis ; Caspase 3 ; Cell Count ; Cell Death ; Cryopreservation ; Diminazene ; DNA ; Fetal Blood ; Transplants ; Trypan Blue

BACKGROUND: In combination with standard-dose CHOP (cyclophosphamide, vincristine, adriamycin, and prednisolone), the addition of rituximab produces a better clinical response in the treatment of aggressive B-cell non-Hodgkin's lymphoma (NHL) than CHOP alone. METHODS: Thirty-four patients with previously untreated diffuse large B-cell NHL received at least three or four cycles of rituximab 375 mg/m2 or 500 mg per dose on day 1 of each cycle in combination with CHOP chemotherapy. RESULTS: The median age of patients were 61.5 years (range, 28-83 years). After the end of therapy, twenty-five patients (73.5%) experienced a complete response, four patients (11.8 %) had a partial response, and two patients (5.9%) were classified as having progressive disease. The median follow-up duration was 9.4 months (range, 0.2-19.5 months) and 1-year overall survival and progression free survival was 84.8+/-8.7% and 80.3+/-9.4%, respectively. Two patients (5.9%) experienced fever, myalgia, and skin eruption due to rituximab. Neutropenia of grade 3 or 4 occurred in thirty-one patients (91.2%). CONCLUSION: The benefits of rituximab in combination with CHOP chemotherapy include high response rates and good tolerance. However, further prospective, randomized studies are needed to draw definitive conclusions.
B-Lymphocytes* ; Disease-Free Survival ; Doxorubicin ; Drug Therapy* ; Fever ; Follow-Up Studies ; Humans ; Lymphoma, B-Cell* ; Lymphoma, Non-Hodgkin ; Myalgia ; Neutropenia ; Skin ; Vincristine ; Rituximab

BACKGROUND: It is still obscure how dendritic cells (DCs) can orchestrate whole immune reactions according to the host age. We studied changes of murine splenic DCs after total body irradiation (TBI), with regards to age. METHODS: Young (8~14 wk) and old (12~16 mo) C57Bl/6 mice were irradiated with a dose of 1,100 cGy and were assessed 6 h later for phenotypic and functional changes of the DCs. The mean fluorescence intensities and cytokine producing cell proportions were analyzed with the student's t-test. RESULTS: Interleukin-12 (IL-12), interferon (IFN gamma) and tumor necrosis factor (TNF alpha) producing classical DCs (cDCs) were more numerous in the young untreated mice than in the old mice. However, the number of these cells decreased in the young mice and increased in the old mice after TBI. IL-12, IFN gamma and TNF alpha producing plasmacytoid DCs (pDCs) were more frequent in the old mice than in the young mice before TBI both mice showed an increased frequency of cells producing these cytokines after TBI. Overall, the highest numbers of cDCs and pDCs producing IL-12, IFN gamma and TNF alpha were present in the old mice after TBI. In both the cDC and pDC populations, the old mice had a higher frequency of IL-10+ cells prior to TBI. After irradiation, the young mice had a higher frequency of IL-10+ cells. CONCLUSION: With TBI, the DCs showed dramatic differences between young and old mice. Young mice turned to an immuno-suppressive response whereas the old mice changed to an immuno-stimulation of DCs after TBI. From these dramatic aging effects, we hope to explain the different frequencies and severities of acute GvHD after allogeneic hematopoietic stem cell transplantation according to host age.
Aging* ; Animals ; Centers for Disease Control and Prevention (U.S.) ; Cytokines ; Dendritic Cells* ; Fluorescence ; Hematopoietic Stem Cell Transplantation ; Hope ; Interferons ; Interleukin-12 ; Mice* ; Tumor Necrosis Factor-alpha ; Whole-Body Irradiation*