PURPOSE: Of the cancers in childhood, leukemia is the most frequent one. For the desirable control of childhood leukemia, the basic data for the incidence has a great importance. The authors made a report about the incidence of leukemia in childhood, which analyzed the data from 126 cases in Kyongnam province, Korea, during 1991~1995. METHODS: The data were obtained from 126 new cases of childhood leukemia who had been living in the Kyongnam province and were diagnosed at the 26 university hospitals or general hospitals in the Kyongnam area and other cities from 1991 to 1995. RESULTS: The age-and-sex adjusted annual incidence rate per 100,000 population during 1991~1995 varied from 1.82 to 2.86, and cumulative annual incidence rate was 2.41 (male 2.26 and female 2.57 respectively). Male to female sex ratio was 1:1 in total cases. By the major types of childhood leukemia, the cases were composed of acute lymphocytic leukemia 70.6%, acute myelocytic leukemia 26.9% and chronic myelocytic leukemia 2.5%. The cumulative annual incidence rate per 100,000 population (crude rate) during 1991~1995 were 2.77 in Ulsan city, 2.62 in Chinju city and 2.34 in the whole area of Kyongnam province. CONCLUSION: It was concluded that the age-and-sex adjusted annual incidence rate per 100,000 of childhood in Kyongnam province was 2.41, which was lower than that in Pusan city in the same period. And, there was no significant difference of the cumulative annual incidence rate between Ulsan area and Chinju area in the same period.
Busan ; Female ; Gyeongsangnam-do* ; Hospitals, General ; Hospitals, University ; Humans ; Incidence* ; Korea* ; Leukemia* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukemia, Myeloid, Acute ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Sex Ratio ; Ulsan

PURPOSE: Autografts with peripheral blood stem cells (PBSCT) have become an accepted alternative to bone marrow transplantation for restoring hematopoiesis after marrow ablative therapy. METHODS: Questionnaires are given to all the transplantation centers in Korea. Eleven centers reported 63 cases and retrospective analysis of these cases were done. RESULTS: Sixty-three children with acute myelogenous leukemia (AML) underwent PBSCT following cytoreductive chemotherapy at 11 transplant centers of the Korean Society of Pediatric Hematology-Oncology from January, 1996 through June, 2000. The patients consisted of 40 males and 23 females with a median age at PBSCT of 10 year (range 1~16). Peripheral blood stem cell (PBSC) were collected by a median of 5 apheresis per patient. Various kinds of multi-drug therapy as cytoreductive regimen were used, and the 'BCVAC' regimen consisting of BCNU, cyclophosphamide, VP-16 and cytarabine was used in 35 patients. The median number of infused mononuclear cell (MNC) and CD34 cells was 8.14 (0.3~26.6) 108/kg and 4.90 (0.12~46.9) 106/kg, respectively. Hematological recovery was evaluated in all patients. The median number of days required to achieve an absolute neutrophil count (ANC) of > 500/mm3, > 1,000/mm3 and platelet count of > 50 103/mm3 was 12 (8~48), 12 (9~84) and 35 (10~370), respectively. Sixteen patients relapsed 1 month to 18 months (median 2 months) after PBSCT, 1 patient progressed to secondary MDS 15 months after PBSCT and 1 patient died at d+39 due to CMV infection. So currently 45 patients are surviving disease free at 2 to 50 months (median 23 months). CONCLUSION: Even though the follow-up period was short and the number of patient was small the autologous PBSCT might be an alternative therapy in childhood AML.
Autografts ; Blood Component Removal ; Bone Marrow ; Bone Marrow Transplantation ; Carmustine ; Child ; Cyclophosphamide ; Cytarabine ; Drug Therapy ; Etoposide ; Female ; Follow-Up Studies ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Humans ; Korea* ; Leukemia, Myeloid, Acute* ; Male ; Neutrophils ; Platelet Count ; Surveys and Questionnaires ; Retrospective Studies ; Stem Cells

The Korean Society of Pediatric Hematology-Oncology surveyed pediatric allogeneic hematopoietic stem cell transplantation in Korea. From 1983 to April 2000, 267 children underwent allogeneic hematopoietic stem cell transplantation. Seventy-nine children were transplanted for acute myelogenous leukemia (AML), 76 for severe aplastic anemia (SAA), 62 for acute lymphoblastic leukemia (ALL), 44 for chronic myelogenous leukemia/myelodysplastic syndrome (CML/MDS) and 11 for nonmalignant rare disease. There were 152 males and 115 females with a median age of 9 years and median follow-up of 25 months. One hundred and eighty-nine of 267 cases were HLA-matched sibling transplants. The estimated event-free survival (EFS) of patients with SAA who underwent HLA-matched sibling transplants is 89%. The estimated EFS of ALL in CR1 and CR2 are 77% and 67%, respectively. The estimated EFS of AML in CR1 and CR2 are 73% and 60%, respectively. The estimated EFS of AML in CR1 prepared with Bu/Cy is 82%. The estimated EFS of CML/MDS is 71%. Eight out of 10 children with nonmalignant rare disease who underwent HLA-matched transplants are alive with disease free. Thirty-three children underwent unrelated bone marrow transplantation and 17 cord blood transplantation. Outcomes of patients with alternative stem cell sources are not estimated due to short median follow-up. These data shows that allogeneic hematopoietic stem cell transplantation is a curative method for children with hematopoietic stem cell disorders and we wish to share these results.
Anemia, Aplastic ; Bone Marrow Transplantation ; Child ; Disease-Free Survival ; Female ; Fetal Blood ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Humans ; Korea ; Leukemia, Myeloid, Acute ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Rare Diseases ; Siblings ; Stem Cell Transplantation ; Stem Cells

Histiocytic necrotizing lymphadenitis (Kikuchi's disease) is a cause of benign lymphadenitis, most commonly affecting young women and usually presenting as a painless or painful cervical lymphadenopathy sometimes associated with fever and leukopenia. Less frequent symptoms include weight loss, nausea, vomiting, and night sweats. We experienced two cases of histiocytic necrotizing lymphadenitis in an 11-year-old boy and a 13-year-old boy. They presented cervical lymphadenopathy with fever in one patient and without fever in the other patient. Lymph node enlargement did not respond to antibiotics. We performed surgical biopsy of cervical lymph node which was consistent with histiocytic necrotizing lymphadenitis. In one patient CD8 T cells and CD68 histiocytes were shown in immunohistochemical stain. So we report two cases of histiocytic necrotizing lymphadenitis with brief review of the related literature.
Adolescent ; Anti-Bacterial Agents ; Biopsy ; Child ; Female ; Fever ; Histiocytes ; Histiocytic Necrotizing Lymphadenitis* ; Humans ; Leukopenia ; Lymph Nodes ; Lymphadenitis ; Lymphatic Diseases ; Male ; Nausea ; Sweat ; T-Lymphocytes ; Vomiting ; Weight Loss

Factor XI deficiency is a very rare autosomal recessive coagulation factor deficiency, comprising 1/million in ethnic groups other than Ashkenazi Jews. The clinical manifestations are extremely variable, and generally milder than those of hemophilia A and B. We describe herewith 3 children with factor XI deficiency, who were found to have prolonged aPTT in routine laboratory studies, or in evaluation of intermittent epistaxis.
Blood Coagulation Factors ; Child ; Epistaxis ; Ethnic Groups ; Factor XI Deficiency* ; Factor XI* ; Hemophilia A ; Humans ; Jews

PURPOSE: CD1a antigen is observed in diseases such as Langerhans cell histiocytosis, T cell acute lymphoblastic leukemia, acute myelogenous leukemia and CD1a presenting cells would be associated with transplantation disorders. In this study, anti-CD1a single-chain Fv (scFv) was made and its DNA sequence was obtained to use as useful implement and informations for the diagnosis, therapy, and study of the diseases mentioned above. METHODS: The cDNA of anti-CD1a scFv was constructed with multiple steps of PCR from NA1/34.HLK and it was inserted into pCANTAB 5 E phagemid. The anti-CD1a scFvs were expressed on transformed DH5alpha Eschericia coli and secreted into cultured media. Thymocytes were used as CD1a antigen presenting cells. ScFvs were tested with flow cytometry. DNA sequence was obtained with PCR generated DNA sequencing method. RESULTS: There were 7 clones that secreted scFvs binding thymocytes. When using thymocytes after being incubated with anti-CD1a antibody, there was only one clone (D-24) scFv of which binding reaction was significantly inhibited. DNA sequence of scFv of D-24 was obtained. CONCLUSION: The obtained scFv of D-24 should be anti-CD1a scFv because of its binding thymocytes and being inhibited by anti-CD1a antibody. It needs further purification and confirmative test such as immunoprecipitation. The obtained DNA sequence would be informative to construct various and humanized antibodies for CD1a antigen.
Antibodies, Monoclonal, Humanized ; Antigen-Presenting Cells ; Bacteriophages* ; Base Sequence ; Clone Cells ; Diagnosis ; DNA, Complementary ; Flow Cytometry* ; Histiocytosis, Langerhans-Cell ; Immunoprecipitation ; Leukemia, Myeloid, Acute ; Mass Screening* ; Polymerase Chain Reaction ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Sequence Analysis, DNA ; Single-Chain Antibodies* ; Thymocytes

PURPOSE: HLA (human leukocyte antigen)-class I genes are highly polymorphic, play many roles in organ and bone marrow transplantation. HLA-B is the most polymorphic class I locus with 414 alleles. HLA-class I typing, which is based on serologic method, has been used until recently. The development of molecular biological techniques make it possible to define the genotypes of HLA genes. METHODS: Analyses of HLA-B genotyping on 1,000 UCB (Umbilical Cord Blood) samples which were considered to be HLA-B homozygote or blank were performed by ARMS-PCR (Amplification Refractory Mutation System-PCR) method and direct sequencing. RESULTS: We could identify HLA-B*5001 which was known to be absent in Koreans. CONCLUSION: It is strongly suggested that HLA-B homozygote should be confirmed to the DNA level especially in cases of donor selection for the unrelated bone marrow transplantation.
Alleles ; Bone Marrow Transplantation ; DNA ; Donor Selection ; Fetal Blood ; Genotype ; HLA-B Antigens ; Homozygote ; Leukocytes

PURPOSE: Current pediatric cancer research requires an organized pediatric tumor tissue bank with standardized guidelines for preparation and storage of human tumor tissue samples, white cells, serum, genomic DNA, RNA, cDNA and proteins.. Our institution established and managed pediatric tumor tissue bank for the last one year, and we want to present an overview of our experiences and guidelines. METHODS: From leukemia patients, peripheral blood and bone marrow aspirates were collected at initial diagnosis. Leukemic cells were prepared by Ficoll density-gradient centrifugation and stored at 196oC liquid nitrogen. For solid tumors, tissue cultures were performed as soon as possible after surgical excision or needle biopsy. Serum free media and primary cultured cells were collected and stored at 20degrees C and at 196degrees C, respectively. Genomic DNA, RNA and cDNA were isolated from leukemic cells and cultured solid tumor cells, and stored at 20degrees C. We also isolated genomic DNA from white blood cells of solid tumor patients and stored at 20degrees C. Finally we collected serum samples from all pediatric cancer patients at diagnosis and stored at 20degrees C. RESULTS: Among the 41 cases of leukemia and 100 cases of solid tumor patients who were diagnosed at department of pediatrics, Samsung Medical Center, from August 2000 to July 2001, 26 cases (63%) of leukemia and 59 cases (59%) of solid tumor patients were registered to Pediatric Tumor Tissue Bank. Primary cell cultures were performed in 21 cases of solid tumors and were successful in 19 cases (90%). The isolated genomic DNA, RNA and cDNA were all in high quality confirmed by electrophoresis in agarose gel. CONCLUSION: The problem of tissue sample size obtained by needle biopsy could be overcome by primary cell cultures. For the effective management of pediatric tumor tissue bank, fresh tissue collection with active cooperation of surgeons, organized personnel structure, and multidisciplinary standardized guidelines are necessary.
Biopsy, Needle ; Bone Marrow ; Cell Culture Techniques ; Cells, Cultured ; Centrifugation ; Culture Media, Serum-Free ; Diagnosis ; DNA ; DNA, Complementary ; Electrophoresis ; Ficoll ; Humans ; Leukemia ; Leukocytes ; Nitrogen ; Pediatrics ; Primary Cell Culture ; RNA ; Sample Size ; Sepharose ; Tissue Banks*

PURPOSE: Dendritic cells (DCs) are the most potent antigen presenting cells and should be differentiated to mature form to induce primary T cell response. In this study, we intended to generate mature DCs from peripheral blood mononuclear cells (PBMCs), so that develope the basis for immunotherapy using DCs. METHODS: PBMCs were isolated from 25 mL of normal adults' peripheral blood and evenly distributed in 5 wells of a 6-well plate. Nonadherent cells were gently aspirated after 2 hour-incubation under humidified 5% CO2 at 37degrees C. Adherent monocytes were cultured in 3 mL of 10% fetal bovine serum plus RPMI-1640 media containing granulocyte/macrophage-colony stimulating factor (GM-CSF) 200 ng/mL and interleukin (IL)-4 20 ng/mL. To assess the effect of tumor necrosis factor (TNF)-alpha and interferon (IFN)-alpha on DC maturation, either or both were added on day 4 of culture. Cells were harvested on day 4 and 7 to calculate the cell counts, CD83 /HLA-DR cells, and CD86 /HLA-DR cells. RESULTS: On day 4, large amounts of DCs were observed. CD83 /HLA-DR cells and CD86 / HLA-DR cells were 11.6% and 16.6% of total cells counted and yields were 1.3% and 2.0%, respectively. On day 7, DCs were more frequently observed in all instances and purity ranged from 24.0% to 31.0% as a mean value. The final yields of matue DCs were 2.9~3.4% of PBMCs inoculated. Adding TNF-alpha plus IFN-alpha led to the best yield. But, IFN-alpha alone did not increase the mature DCs compared to the control. CONCLUSION: We successfully cultured large quantities of mature DCs from PBMCs using GM-CSF and IL-4. IFN-alpha seems to have a synergistic effect when added with TNF-alpha, but further studies are required to prove the clinical significance.
Antigen-Presenting Cells ; Cell Count ; Dendritic Cells* ; Granulocyte-Macrophage Colony-Stimulating Factor ; HLA-DR Antigens ; Immunotherapy ; Interferon-alpha ; Interferons ; Interleukin-4 ; Interleukins ; Monocytes ; Tumor Necrosis Factor-alpha

PURPOSE: Cryopreservation of hematopoietic stem cells is one of the essential components in autologous and peripheral blood stem cell transplantation. Cryopreservation of hematopoietic stem cell, the conventional method involves controlled-rate freezing by a programmed freezer in medium that contains 10% dimethyl sulfoxide (DMSO) as cryoprotectant, followed by storage in liquid nitrogen freezer. We compared the differences between different methods of cryopreservation and cryoprotectants on viability and colony forming capacity of hematopoietic stem cells. METHODS: Mononuclear cells separated using Ficoll-Hypaque from cord blood, peripheral blood and bone marrow were frozen with programmed freezer at 196degrees C or placed in a 70degrees C freezer without programmed freezer in both 10% and 20% DMSO. We measured cell viability using trypan blue dye exclusion method and colony forming capacity with methyl cellulose media at 7, 30 and 90 days after thawing. RESULTS: Cell viability of cord blood, peripheral blood and bone marrow was higher in the groups with programmed freezer compared with rapid freezing and storing in a 70degrees C freezer. Also as the storage time passed, the decrease in viability of hematopoietic cells was much less in the groups of controlled-rate freezing by a programmed freezer. The number of colony in cord blood and bone marrow was higher with programmed freezer and that of peripheral blood was higher with rapid freezing and storage in a 70degrees C freezer. Comparing the differences between different concentraions of DMSO, cell viability was similar or slightly higher in 20% DMSO groups than 10% DMSO groups, but the number of colony was higher in 10% DMSO groups. CONCLUSION: These results suggested that conventional cryopreservation method using programmed freezer with 10% DMSO was more effective in the cryopreservation of hematopoietic stem cells.
Bone Marrow ; Cell Survival ; Cryopreservation* ; Dimethyl Sulfoxide ; Fetal Blood ; Freezing ; Hematopoietic Stem Cells* ; Methylcellulose ; Nitrogen ; Peripheral Blood Stem Cell Transplantation ; Stem Cell Transplantation ; Trypan Blue