Establishment and Characteristics of the B Cell Line (KEB1) from the Bone Marrow Cells of the Patient Infected with Epstein-Barr Virus / 대한혈액학회지

BACKGROUND: Cell lines can be established when the cells are clonally selected and propagated continuously in vitro culture system. Recently we established a B cell line (KEB1) from the bone marrow cells of the patient infected with Epstein-Barr virus (EBV). METHODS: The patient's initial platelet count was 1,000/microliter and peripheral blood smear showed atypical lymphocytes accounting 20% of the differentials of WBC. Antibodies to EBV and PCR for EBV were positive but heterophil antibody was negative. Mononuclear cells were obtained by Ficoll-paque separation and suspended in RPMI media with 10% FCS. After incubation in 37degrees C, 5% CO2 incubator, cells grew continuously and finally immortalized to B cell line. RESULTS: Cells showed abundant, clear basophilic cytoplasms and a few vacuoles. Cells had granular reaction in PAS stain and were positive to B cell antibodies. Immunohistochemical stain showed positive expression for EBV antibody. Electron microscopic finding of cultured cells showed several viral particles, and immunoelectron microscopic finding showed electron dense expression. Immunophenotyping of cultured cells was positive for B lymphoid lineage, and karyotypings had hypotetraploidy. Cells expressed MAGE and SSX gene. Cytotoxicity showed relative resistance to mistletoe and several chemotherapeutic agents compared to leukemic cell line. CONCLUSION: KEB1 cell line was established from the bone marrow cells of the patient with infectious mononucleosis. The characteristics of the cell lines including morphology, immunophenotype, karyotype, gene analysis (MAGE, SSX) and chemosensitivity were analyzed. There should be further studies of these cell lines including gene analysis, telomerase activity and cytokine production. This cell line might be helpful to establish another normal lymphocyte cell line and to predict the toxicity of chemotherapy.

Four Cases of Staphylococcal Scalded Skin Syndrome / 소아과

Staphylococcal scalded skin syndrome is a systemic disease with a clinical spectrum ranging from subcorneal pustules, patterned exfoliation to extensive erosion and peeling of skin by the exfoliative toxin of group II Staphylococcus aureus. This disease occurs mainly in infancy and children below five years and it isn't easy to differentiate from other vesicular diseases clinically, but skin biopsy shows an epidermal split at the granular layer. The form and severity of staphylococcal scalded skin syndrome will vary according to defense system and toxic factors. Treatment is effective antibiotics, and the mortality rate increases to 5% in children. In our four cases, symptoms were erythema and fever, followed by formation of large bullae and denuded skin. On laboratory findings, leukocytosis was noted in three cases, and S. aureus was confirmed by culture of eye discharge in all cases. Our cases improved with antibiotic therapy. We experienced four cases of staphylococcal scalded skin syndrome which were presented with vesicle and exfoliative skin lesion and treated successfully.

Endoscopic Findings of Children with Henoch-Schonlein Purpura

PURPOSE: This study was performed to analyze the endoscopic findings in Henoch-Schonlein purpura patients, and to compare the differences in endoscopic findings according to age and gastrointestinal symptoms. METHODS: We examined children with Henoch-Schonlein purpura aged 3 to 15 years between September 1996 and October 2002. The total number studied was 65, consisting of 41 boys and 24 girls. Endoscopy was performed and the results were analysed. RESULTS: Among 65 cases, 12 cases of duodenitis, nine cases of gastritis and duodenitis, six cases of duodenal erosion, five cases of gastritis, five cases of duodenal ulcer, two cases of gastric ulcer and one case of colonic erosion were noted. Endoscopic abnormality was found in 38 of 53 who had gastrointestinal symptoms, and in two of 12 who didn't have gastrointestinal symptoms. CONCLUSION: Most of the gastrointestinal symptoms in Henoch-Schonlein purpura patients were relieved without complication. But in some cases severe symptoms such as hematemesis, melena, and abdominal pain localized to epigastric area were developed when diagnosis was delayed. Prompt endoscopy will be helpful for diagnosis and therapy of Henoch-Schonlein purpura with gastrointestinal involvement.

MRI Finding of a Case of Relapsed Acute Mixed Type Leukemia with Bone Marrow Necrosis

Bone marrow necrosis is a rare complication of a variety of diseases affecting the marrow. The cause and incidence are unknown, and reports of treatment response are rare. We describe a case of relapsed acute mixed type leukemia with bone marrow necrosis. The patient was a 10 year old female diagnosed with acute mixed type leukemia four years ago. She had been on second remission state for 1 year, presented with severe back pain, tenderness in lower extremities, low-grade fever and general weakness. Her level of serum lactic dehydrogenase on admission was increased. Bone marrow aspiration from both posterior iliac crest showed marrow necrosis. Subsequent examination showed the same feature. Hip MRI showed heterogenous low signal intensity in both iliac bone on T-1 weighted image and heterogenous high signal intensity on T-2 wieghted image. Remission induction therapy was started but she expired on 59th hospital day due to the complication of sepsis.

Detection of N-myc Amplification with Differential PCR in Neuroblastoma and It's Clinical Significance / 대한소아혈액종양학회지

PURPOSE: The N-myc amplification is one of well known poor prognostic markers in neurblastoma. Because the traditional detection method, Southern blot, is expensive, labor-intensive and time-consuming, the detection of N-myc amplification is not routinely performed in Korea. The purposes of this study are to develop polymerase chain reaction (PCR) for detecting N-myc amplification in neuroblastoma tumor tissue, and to elucidate the clinical significance of N-myc amplification. METHODS: The clinical data and paraffin embedded tumor specimen of 54 neuroblastoma cases were collected from 10 medical centers in Korea. We have developed semiquantitative method of estimating gene copy number that uses differential PCR. N-myc gene primers (RC N-myc, N-myc 7-1) are amplified together with primers from a single-copy internal control gene (beta-globin). After ethidium bromide-stained agarose gel electrophoresis, the ratio of the two PCR products, which stands for N-myc amplification, is determined. Kaplan-Meier survival analysis was performed to evaluate the prognostic significance of N-myc amplification. RESULTS: The differential PCR was very effective, less expensive, less labor-intensive, and simple detection method for N-myc amplification. The percentage of N-myc amplification was higher in the patients older than 1 year old (34.1%: 14/41), when they were compared to the patients younger than 1 year old (16.7%: 2/12). The percentage of N-myc amplification was higher in the patients who have primary tumor at adrenal gland (40.9%: 9/22) than who have primary tumor at retroperitoneum (17.6%: 3/17) or at mediastinum (16.7%: 2/12). In Stage I, II, and III patients, the mean survival time of N-myc amplified group was 18 months and that of N-myc umamplified group was 64 months (Log Rank 4.35, P=0.037). CONCLUSION: The differential PCR was very effective, less expensive, less labor-intensive, and simple detection method for N-myc amplification. The N-myc amplification is one of poor prognostic indicators in Neuroblastoma.

Viability and Colony Forming Capacity of Hematopoietic Stem Cells after Cryopreservation / 대한소아혈액종양학회지

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.

Infectious Manifestations in Childhood Leukemia

PURPOSE: Infection is one of the major causes of morbidity and mortality in children during chemotherapy for leukemia and the development of fever in neutropenic cancer patients frequently indicates infection. The purpose of the present study is to evaluate infectious manifestations during the course of leukemia. METHODS: Seventy eight leukemic children who had one or more occasions of infection during hospitalization from January 1993 to December 1999 at Dong San Hospital, Keimyung University were analyzed. Infection was defined clinically as a single oral temperature of 38.5degrees C or higher or as three consequent oral temperature 38degrees C in a day. RESULTS: Two-hundred and four febrile episodes were studied. The cause of infection was detected in 136 episodes while in 68 episodes the cause was not detected. The causes of infection were : 31 pneumonia, 24 sepsis, 16 urinary tract infections, 15 mucositis, and 14 wound infections. The etiologic pathogens were identified in 53 episodes. Sixty percent of the pathogens were gram negative organisms such as Escherichia coli, Enterobacter, Pseudomonas aeruginosa and Klebsiella. Both gram positive and fungal infections were 17.0%. Most sensitive antibiotics were vancomycin for gram positive organisms and ceftazidime and amikacin for gram negative organisms. Twenty-one patients died due to FUOs, sepsis, pneumonia and severe mucositis. CONCLUSION: Infection was the most frequent cause of death in leukemic patients and fungal infections have increased recently. The risk of infection was higher in patients with severe and prolonged neutropenia. Therefore immediate application of antibiotics and antifungal agents will be needed in the leukemic patient with neutropenia.

A Case of Acute Leukemia Remitted by Adding Cyclosporin-A Previously Failed with Induction Therapy

Clinical chemotherapy refractoriness is characterized by resistance to multiple drugs. Multidrug resistance(MDR) is caused by over-reactivity of a unidirectional drug efflux pump, transmembrane glycoprotein(P-glycoprotein), which is encoded by the MDR1 gene. P-glycoprotein leads to increased drug efflux and decreased intracellular drug concentration. Clinical trials that attempt to reverse or modulate MDR have been done. Cyclosporin-A and verapamil are the most extensively studied agents and several trials of cyclosporin-A as a MDR modulator have been reported. We report a case of an 8-year-old girl with acute mixed type leukemia who failed to respond 3 times to remission-induction therapy. It led us to conclude she had multidrug resistance. We tried a fourth induction chemotherapy including cytarabine, idarubicin and 6-thioguanine to which cyclosporin-A was added. Then, she showed signs of severe bone marrow depression and fulminant perianal cellulitis. But she recovered and successfully achieved complete remission. The addition of cyclosporine could be useful in achieving complete remission for cases of acute leukemia that resist to usual chemotherapy. Futher observation including more cases will be needed to assess long-term survival and efficacy of adding cyclosporine.

Thrombopoietin Level and Bone Marrow Megakaryocyte Colony Formation in Various Diseases with Thrombocytopenia / 대한소아혈액종양학회지

PURPOSE: Thrombocytopenia is a serious life threatening consequence in patients with bone marrow failure syndrome. Thrombopoietin (TPO), recently cloned by several groups has been shown to be a key regulation of megakaryopoiesis and thrombopoiesis. Recent studies have demonstrated a positive or negative relationship between TPO levels and platelet counts due to underlying disease states. To clarify the role of TPO in thrombocytopenic condition we determined plasma TPO levels and megakaryocyte colony assay. METHPDS: TPO levels were measured in thrombocytopenic patient with aplastic anemia, chemotherapy induced bone marrow failure, idiopathic thrombocytopenic purpura (ITP) and in newborn by ELISA (QuantikineTM, R&D System, USA). Controls were short statured normal children with normal platelet counts. Plasma was preserved in 20oC until test. CFU-mega was determined by MegaCultTM (Stem Cell Tech. Inc., Canada). Ficoll separated mononuclear cells were cultured for 10~12 days with TPO or stem cell factor (SCF) in 37degrees C 5% CO2 atmosphere, colonies were fixed, stained and examined with inverted microscope. Results were analysed by Student-t test. RESULTS: TPO levels were markedly increased in aplastic anemia and chemotherapy induced thrombocytopenia compared to those of normal controls. Patients with ITP had decreased level of plasma TPO. There was inverse relationship between platelet count and TPO levels for patients with aplastic anemia and chemotherapy induced thrombocytopenia. There was no definite relationship between platelet counts and TPO levels but inverse relationship between platelet counts and PDW levels in neonates was noted. The levels of TPO were increased after improvement of platelet in thrombocytopenic neonate. Megakaryocyte colonies were increased in the mononuclear cells of the patients with ITP and chemotherapy induced thrombocytopenia. There was little colony formation in aplastic anemia. TPO had no definite effect in megakaryocyte colony formation but SCF increased colony formation. CONCLUSION: TPO levels were increased in aplastic anemia and chemotherapy induced thrombocytopenia but decreased in ITP. There was inverse relationship between platelet count and TPO levels in aplastic anemia and chemotherapy induced thrombocytopenia. Thus TPO could be useful for differentiate the etiology of thrombocytopenia. Megakaryocyte colony was increased in ITP and chemotherapy induced thrombocytopenia, but decreased in aplastic anemia. SCF was effective in megakaryocyte colony formation. TPO and SCF will be helpful to increase platelet in thrombocytopenic patients. However, further study will be needed.

A Case of Relapsed Acute Promyleocytic Leukemia Induced Remission with Arsenic Trioxide(As2O3)

Acute promyelocytic leukemia(APL) is a unique entity in the spectrum of acute myelogenous leukemia. It has several characteristic features, including distinctive morphology, chromosomal translocation, t(15:17), disseminated intravascular coagulation and effect on retinoic acid. Retinoic acid which is a derivative of vitamin A induces differentation of APL cells in vitro and in vivo, but its cessation induces relapse of APL. Arsenic trioxide(AszOz) can induce clinical remission in patients with APL, including those who have relapsed after retinonic acid treatment. We report a case of a 9-year-old male with APL who had relapsed after cessation of retinoic acid treatment. The patient successfully achieved remission following treatment with AsO. Arsenic trioxide treatment would be an effective and relatively safe drug in childhood APL patients refractory to retinoic acid.

Clinical Manifestation of Childhood Acute Leukemia with Bone Involvement

PURPOSE: Bone involvement is known to develop in 40-70Yo of pediatric acute leukemia. We aimed to analyze the clinical course and result of therapy in pediatric acute leukemia with bone involvement. METHODS: Twenty-seven patients diagnosed as pediatric acute leukemia at Dong San Medical Center from Jan. 1996 to Aug. 1998 were evaluated. According to bone X-ray and whole body bone scan, the patients were divided into two groups. RESULTS: Twenty-seven patients were enrolled in this study with 14 patients(52Yo) showing definite bone involvement on simple X-ray or bone scan. Mean age of patients with bone involvement was 5.5 years. Regarding the type of leukemia, 9 patients(64%) were acute lymphocytic leukemia. Ten patients(71%) out of 14 with bone involvement complained of bone pain at the involved bony site. Site of involvement was most frequent in the lower extremity. On simple X-ray, osteolytic lesion was found in 7 patients(50%), diffuse osteopenia in 2 patients(14%) and pathologic fracture in 2 patients(14%). In bone scan, radioactivity was increased in whole cases of patients with bone involvement. Thirteen patients(93%) were completely remitted by chemo-therapy, but, one AML patient died due to induction failure. CONCLUSION: Bone involvement occured in 52% of pediatric acute leukemia. Bone involvement was more frequent in male patients in the lower extremity, and osteolytic lesion was the most frequent finding on simple X-ray. There was no relevence between bone involvement and prognosis. Further study will be needed to evaluate long-term survival and prognosis. (J Korean Pediatr Soc 2000;43:806-813)

Childhood Cancer and Infection

No abstract available.

Anticancer Effect of Arsenic Trioxide in Acute Promyelocytic Leukemia

PURPOSE: Acute promyelocytic leukemia (APL or AML, M3) represents an unique model for cancer research in terms of biological and clinical features. Since 1988, it has been widely confirmed that all-trans retinoic acid (ATRA) can induce complete clinical remission in over 85% of APL patients by a differentiation process, with PML-RARalpha protein possibly being the direct target of ATRA. However, ATRA treatment has two clinical limitations, namely, retinoic acid syndrome and retinoic resistance. Recently, it has been shown that arsenic trioxide used in some traditional Chinese remedy is very effective in retinoic resistant APL treatment. We tried to observe arsenic effect on cell lines and APL patient cells. MEHTODS: We investigated arsenic trioxide-induced apoptosis on APL, HL60, K562, KPH1 cell lines through MTT assay, DNA fragmentation assay and morphologic features. RESULTS: In MTT assay, cell survival rate decreased as the concentration of arsenic trioxide increased. In DNA fragmentation assay with HL60 cell line, DNA fragmentation was more frequent in high concentrations of arsenic trioxide than in low concentrations. During arsenic trioxide treatment, the morphologic change in bone marrow cells of APL patient, included nuclear differentiation and dark cytoplasmic granule during arsenic trioxide treatment. Serum arsenic reached peak level at 4hr after injection. We experienced a case of a 9-year-old male with APL who had relapsed after cessation of retinoic acid treatment. The patient successfully achieved remission following arsenic trioxide treatment without bone marrow depression and exacerbating bleeding diathesis. CONCLUSION: Arsenic trioxide can be used effectively to treat APL patients by inducing apoptosis and partial differentiation in tumor cells. The precise cellular and molecular mechanisms of its therapeutic effects remain to be determined.

Hematopoietic Stem Cell Colony Formation of the Fetal Liver / 대한혈액학회지

BACKGROUND: Hematopoietic stem cells of the human fetal liver prior to 15 weeks gestation have remakable advantages for successful engraftment due to embryological immune immaturity, especially in-utero transplantation. This study was undertaken to obtain objective assessment data about the possibility of fetal liver hematopoietic stem cell transplantation in the future. METHODS: Six cases of the fetal liver tissue were obtained from therapeutic abortions at 12~20 weeks gestation. The fetal liver was collected in RPMI media containing 10% fetal calf serum and the cell suspensions were obtained by centrifugation following physical disruption. The number of nucleated cells in each case was counted and the colony numbers in methyl cellulose media were scored according to incubation period with or without growth factors. Some of the cells were cryopreserved in the liquid nitrogen tank, thereafter cell viability and colony numbers were evaluated according to cryopreservation period. RESULTS: The nucleated cell numbers obtained from each fetal liver increased with gestational age. The colony numbers after incubation increased with gestational age and the erythroid lineage was predominant in 3 cases which are under 15 weeks gestation. The colonogenic activity after incubation with combination of hematopoietic growth factors increased in only one case. The cell viability and the colony numbers after cryopreservation was decreased compare to the value before cryopreservation. CONCLUSION: The number of nucleated cells and hematopoietic stem cell colony formation were increased with gestational age and viability of the cells after cryopreservation was decreased. Further systematic studies using more cases would be needed to obtain objective assessment data for fetal liver transplantation program in the future.

A Clinical Study of Childhood Histiocytosis Syndrome / 대한소아혈액종양학회지

PURPOSE: We surveyed this study to find the factors related to clinical aspects of patients with histiocytosis syndrome. METHODS: We analyzed the clinical data of thirty patients retrospectively who were diagnosed as histiocytosis syndrome from January 1992 to December 1997 at Keimyung University Dong San Hospital. RESULTS: There were nine cases of Class I patients, twenty cases of Class II patients and one case of Class III patient. Male patients were eighteen, and female patients were twelve. Mean age at diagnosis was 4 years. The most common clinical manifestation was fever, and others were hepatosplenomegaly, pallor, respiratory symptom, and lymphadenopathy in order. Bone was involved in seven cases out of nine Class I patients. Single organ involvement happened in five cases out of Class I patients, two organ involvement happened in two patients, three or four organ involvement happened in one case of Class I patient respectively. Etiology of Class II were EBV in four patients, bacterial infection in four patients, and the others were candida, mycoplasma, mycobacterium tuberculosis. There were pancytopenia, coagulation defect, abnormal liver function tests on laboratory examinations. Most common histologic finding of Class I was proliferation and infilteration of histiocytes. Hemophagocytosis was common in bone marrow examination of Class II patients. Chemotherapy was undergone for seven patients out of nine Class I patients. Six of them showed complete remission. One of them died during chemotherapy. Thirteen patients out of twenty Class II patients are on complete remission, and five of them died. One Class III patient died during chmotherapy. CONCLUSION: The survival rate depends on age, Lahey's organ dysfunction score, severity, and sites of involved organ. One year survival rate by Kaplan-Meier method of ClassI and II patients was 87.5% and 72.2% respectively. In this study, Class II patients showed high mortality rate, so early diagnosis and treatment will be important.

Epidemiologic Investigation of Childhood Cancer in Taegu, Kyungpook Area / 대한소아혈액종양학회지

PURPOSE: This study was carried to examine the temporal trend and geographical distribution of the childhood cancers in Taegu and Kyungpook province and to postulate an etiological hypothesis for development of the childhood cancer. METHODS: A total of 799 childhood cancer patients whose addresses were either Taegu or Kyungpook province were diagnosed at 5 major hospitals in Taegu from January 1982 to December 1996. The types, sexes, years, and frequencies of the childhood cancer and regional distributions were analyzed, based on the hospital records of these patients. RESULTS: The most common childhood cancer was leukemia that accounted for 49.2% of all childhood cancer cases and it was followed by CNS tumor (12.3%), lymphoma (8.4%), neuroblastoma (7.4%), Wilms tumor (3.9%), retinoblastoma (3.4%), rhabdomyo sarcoma (2.7%), bone tumor (2.4%), embryonal carcinoma (1.9%), hepatoblastoma (1.3%) and others (7.1%). Male to female ratio of the cases was 1.5:1. The changes of the annual incidence rates over 15 years in Taegu and Kyungpook area were not consistently increasing but rather variable. Cancer incidence rate of Taegu was significantly higher than that of Kyungpook province (P<0.005). The incidence rates of industrialized cities around Taegu were significantly higher than those of agricultural regions of northern Kyungpook (P<0.05). CONCLUSION: Geographical difference in cancer incidence rate suggested that certain environmental factors may be associated with the childhood cancer. To identify such factors an analytical epidemiologic study is warranted. For the analytical epidemiologic study, a detailed history of residential area and occupational history of parents should be recorded uniformly for all the new childhood cancer cases.

Chemosensitivity Test of Childhood Leukemic Cells

PURPOSE: The purpose of the present study is to determine the relation between in vitro resistance to 9 drugs, measured with colorimetric methylthiazol tetrazolium(MTT) assay and prognostic factors. METHODS: Thirty children with leukemia were evaluated at the pediatric department of Dongsan Medical Center. All samples tested with the MTT assay contained 80% or more leukemic cells, which were isolated by Ficoll density gradient centrifugation, and were incubated with 9 drugs for 4 days. The optical density(OD) of the wells was measured with microplate spectrophotometer. Leukemic cell survival(LCS) was calculated by OD treated well/OD control wellsx100(%). LD50 was calculated from the dose-response curve and used as a measure of resistance. RESULTS: Among the 30 children with leukemia, 16 were ALL, 14 were AML. Seventeen boys and thirteen girls ranged in age from 9 months to 16 years. Comparing LD50 values according to leukemic type, AML revealed relatively high LD50 values for all drugs, except VCR. But there were no significant differences between ALL and AML(P>0.05). Male showed high LD50 values for ASP, DET, ARA-C, VP16, ADR and 6TG. Age10 years children showed high LD50 values for all drugs, except 6TG. Patients with a leukocyte count>100,000/mm3 at diagnosis showed high LD50 values for VCR, ASP, DET, MTX, ARA-C, ADR, and 6TG. Patients with normal chromosome showed higher LD50 values. CONCLUSION: Our study showed higher LD50 values at AML, male, ageyears old, leukocyte count>100,000/mm3, and normal karyotype. The MTT test may contribute to the selection of effective chemotherapeutic agent for children with acute leukemia.

Surgical Treatment for Intractable Childhood Epilepsy

PURPOSE: For certain forms of childhood epilepsy that remain uncontrolled despite adequate treatment with standard antiepileptic medication, surgical therapy should be considered as a potential treatment. The prognosis for seizure control after early surgery is favorable and is at least comparable with that of adults. With the exception of the obvious benefit conferred by alleviating seizures at a younger age, early surgery also later improves psychosocial status and adaptive function. This study was performed to evaluate the efficacy of epilepsy surgery. METHODS: We analyzed the results of 28 cases of intractable childhood epilepsy who underwent epilepsy surgery at the epilepsy center of Dongsan Medical Center between February, 1993 and January, 1996. They followed up for at least 15 months after surgery. Seizures began at 14 days to 15 years (mean 6.3 years) after birth and had been refractory to antiepileptic medications. Presurgical evaluations of epilepsy included detailed clinical history, scalp/sphenoidal EEG, Video-EEG monitoring, neuroimaging, neuropsychological test, Wada test and invasive study with subdural electrodes. RESULTS: Temporal lobectomy (with or without corticectomy) was performed in 13 cases, extratemporal lobectomy in 11 cases (frontal lobe n=7, parietal lobe n=2, frontoparietal n=1, parietooccipital n=1), functional hemispherectomy in two cases and corpus callosotomy in two cases. The surgical outcome was better in temporal lobe epilepsy compared with that of extratemporal lobe epilepsy. In temporal lobe epilepsy, seven of 13 cases had class I outcome grade, four cases had class II and the rest had class III and class IV. In extratemporal lobe epilepsy, five of 11 cases had class I outcome and the remainders had class III, IV. CONCLUSION: Our results agree with previous reports that epilepsy surgery can provide relief from intractable seizure in pediatric patients, but more extensive study for the patients' cognitive and behavior status will be necessary.

Megakaryocyte Colony Formation According to the Origin of Hematopoietic Stem Cell / 대한소아혈액종양학회지

BACKGROUND: Development of hematopoietic growth factor made it possible to treat anemia and granulocytopenia following intensive chemotherapy and for thrombocytopenia, recently found thrombopoietin(TPO) is being applied experimentally in several countries. The megakaryocyte colony assay can assess the effect of TPO on the thrombocytopenia resulted from cancer chemotherapy or hematopoietic stem cell transplantation. In vitro colony assay procedures for detecting human erythroid and granulocyte macrophage progenitors have been in widespread use for many years. However, reproducible assay methods for human megakaryocyte progenitors have lagged considerably behind especially in Korea. Duration platelet recovery following transplantation depends on the origin of the hematopoietic cells. Usually thrombocyte recovery is delayed following cord blood stem cell transplantation because of the small amount of cells administered. This study was carried out to investigate and establish the megakaryocyte colony assay of hematopoietic stem cells obtained from the various origin of the hematopoietic stem cells with or without TPO. METHOD: Mononuclear cells of bone marrow, peripheral blood and cord blood were collected following Ficoll density gradient centrifugation and megakaryocyte colony assay was done using MegaCultTM(Stem Cell Tech. Inc., Canada). After liquifying the agarose, mononuclear cells were added and then agarose and cell mixture were dispersed into the two wells of the chamber slide. These slides were incubated for 18~21 days at 37oC, 5% CO2. The megakaryocyte colonies were detected by staining of the cells with a primary antibody to the GPIIb/IIIa antigen, secondary antibody, alkaline phosphatase and Evans Blue in order. Changes of CD34 and GPIIb/IIIa positive cells were also analysed in flask culture using flow cytometry. RESULTS: CD34 positive cells were most abundant in the mononuclear cells of the bone marrow, meanwhile the number of CFU-GM and megakaryocyte colony were greater in the mononuclear cells of the cord blood. After administration of TPO, the cell number of megakaryocyte colony was increased dose dependently, but CFU-GM colony did not show any response to TPO. With flask culture, the cell number was decreased with or without TPO. However adding GM-CSF, IL3 and TPO to cord blood mononuclear cell, the number of the cord blood mononuclear cells was increased on the 5 th day. The amount of CD34 positive cells was increased dose dependently to TPO in one of two cord blood and one peripheral blood. The amount of GPIIb/IIIa positive cells was increased dose dependently to TPO following incubation of all the mononuclear cells. CONCLUSION: This study revealed successful result of megakaryocyte colony assay using MegaCultTM in various kinds of mononuclear cells and suggested that TPO was useful for CFU-mega colony formation. The amount of GPIIb/IIIa positive cells was increased with TPO in the flask culture. Therefore TPO could be useful for assessment of CFU- mega, and could be applied for the in vivo and in vitro expansion of megakaryocytes and platelets.

Change of Thyroid Function during Chemotherapy in Chilolren with Acute Leukemia / 대한소아혈액종양학회지

BACKGROUND: The overall prognosis of acute leukemia has dramatically improved in the past 20 years, primarily due to the use of intensive multiagent chemotherapy in combination with CNS prophylaxis. However, increased aggressiveness of treatment protocols was entailed a great risk of various toxic effects. Endocrine function was also affected. The aim of this study is to compare the effect of chemotherapy on thyroid function in children with acute leukemia. METHOD: Parameters of thyroid function during chemotherapy were measured in 11 children with acute leukemia. Level of the serum 73,74 and TSH were determined before therapy, 7th day and 30th day of chemotherapy. Determination of serum 73, 74 and TSH were performed by conventional radioimmunoassay technique. Statistical analysis was done using SAS software. RESULT: 1) Level of level 73 was normal in 7 cases before therapy and decreased in 9 cases on 7th day of remission induction therapy. On 30th day, 73 level was increased to normal value. 2) Level of 74 was normal before therapy and decreased on 7th day of therapy. On 30th day of therapy 74 level showed various change. Three of them showed sustained low level of 74 on 7th and 30th day. 3) Level of TSH were normal before therapy and decreased on 7th day of therapy, followed achievement of normal level after completion of induction therapy. CONCLUSION: We conclude that during induction chemotherapy in childhood acute leukemia, thyroid function was impaired which was reversible.