• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2153-2158
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• 2014

Beclin 1 is a key factor for initiation and regulation of autophagy, which is a cellular catabolic process involved in tumorigenesis. To investigate the role of alternative splicing of Beclin1 in the regulation of autophagy in leukemia cells, Beclin1 mRNA from 6 different types of cell lines and peripheral blood mononuclear cells from 2 healthy volunteers was reversely transcribed, subcloned, and screened for alternative splicing. New transcript variants were analyzed by DNA sequencing. A transcript variant of Beclin 1 gene carrying a deletion of exon 11, which encoded a C-terminal truncation of Beclin 1 isoform, was found. The alternative isoform was assessed by bioinformatics, immunoblotting and subcellular localization. The results showed that this variable transcript is generated by alternative 3' splicing, and its translational product displayed a reduced activity in induction of autophagy by starvation, indicating that the spliced isoform might function as a dominant negative modulator of autophagy. Our findings suggest that the alternative splicing of Beclin 1 might play important roles in leukemogenesis regulated by autophagy.

• IMMUNE NETWORK
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• v.10 no.6
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• pp.239-246
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• 2010

Background: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. Methods: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. Results: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. Conclusion: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.

• The Journal of the Korean Society for Microbiology
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• v.13 no.1
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• pp.1-5
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• 1978

Listeria monocytogenes infection was considered a rather rare disease and occurs mostly either in newborn babies or in young children. However, there has been increasing reports of this infection in elderly person with various underlying disease. Recently we have experienced two cases of Listeria meningitis; one in a 4-year-old male with an acute lymphoblastic leukemia, and the other in a 43-year-old female with a breast cancer. Both were on various chemotherapeutic agents for their primary diseases when the organism, L. monocytogenes was found in their celebospinal fluid(CSF). The degree of CSF pleocytosis were quite different by cases. The former case showed a marked increase, $3,350/mm^3$, and the latter slight, $410/mm^3$, Both showed a slight decrease of CSF glucose ranging 39 to 43mg/100ml. It seems that a routine CSF analysis bears a limitted value in the diagnosis or Listeria meningitis. A direct smear of CSF with Gram's stain revealed gram-positive bacilli in one case, but none in the other. Bacterial culture of CSF yielded plenty colonies in one case, but a few in the other. It seems that isolation of L. monocytogenes must not be considered very easy, and a negative direct smear does not necessarily mean a negative culture. The two isolates we obtained showed the typical cultural and biochemical characteristics of L. monocytogenes and were found to belong to serotypes 1b and 4b. It was our experience that the identification of this organism was not very much matter because of its distinct characteristics, but the most important matter was how to think of the possibility of this organism at the begining. The two isolates were both susceptible to cephalothin, chloramphenicol, erythromycin, tetracycline and gentamicin; intermediate to ampicillin, penicillin and kanamycin; and resistant to cloxacillin.

• Korean Journal of Head & Neck Oncology
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• v.17 no.1
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• pp.26-31
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• 2001

Background: Primary malignant lymphoma of the parotid gland is a rare disease and defined as any malignant lymphoma that first manifests in the parotid gland, regardless of the subsequent stage of the diseases, whether it arises in the parenchyma or intraglandular lymph nodes. This study was performed to review the clinicopathological characteristics of primary parotid lymphoma and identify its optimal treatment modality. Materials and Methods: Six cases with parotid mass as first presentation of malignant lymphoma between 1988 and 2000, were studied on the basis of clinical features, diagnostic tools, treatment modality, treatment outcomes, and clinical stage by Ann Arbor Criteria. All were microscopically reevaluated and classified by NCI working formulation. Results: All patients were males and mean age was 36.7 years (2-66 years). Rapid growing non-tender mass was presented in all the cases and cervical lymphnodes were palpated in 4 cases. However, there was not any evidence of concurrent autoimmune disease such as Sjogren's syndrom or Rheumatoid arthritis. One case was confirmed by surgical specimen after superficial parotidectomy, 2 by excisional biopsy, and 3 by incisional biopsy. The stage of disease by NCI working formulation was IE in 1 patient, IIE in 4 and IV in 1. All were classified into non-Hodgkin' lymphoma, of which there were 5 cases of B-cell type and 1 case of T-cell type. There were 3 diffuse large cell lymphomas, 1 Burkitt lymphoma, 1 MALT lymphoma and 1 T-lymphoblastic lymphoma. Three cases were treated by chemotherapy only, 2 by radiotherapy only and 1 by chemo-radiotherapy. One case with Burkitt lymphoma was died from the disease and one case was lost to follow-up. The others are alive with no evidence of recurrence. Conclusions: Although primary parotid lymphoma is rare and difficult to diagnose preoperatively, most were detected in early stage and showed a relatively good response to the chemotherapy or radiotherapy like other types of extranodal malignant lymphoma.

• Archives of Pharmacal Research
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• v.29 no.10
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• pp.866-873
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• 2006

When patients with cancers are treated with chemotherapeutic agents a long time, some of the cancer cells develop the multidrug resistance (MDR) phenotype. MDR cancer cells are characterized by the overexpression of multidrug resistance1 (MDR1) gene which encodes P-glycoprotein (Pgp), a surface protein of tumor cells that functions to produce an excessive efflux and thereby an insufficient intracellular concentration of chemotherapeutic agents. A variety of studies have sought potent MDR modulators to decrease MDR1 gene expression in cancer cells. Our previous study has shown that curcumin exhibits characteristics of a MDR modulator in KB-V1 multidrug-resistant cells. The aim of this study was to further investigate the effect of curcumin on MDR1 gene expression in patient leukemic cells. The leukemic cells were collected from 78 childhood leukemia patients admitted at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand, in the period from July 2003 to February 2005. There were 61 cases of acute lymphoblastic leukemia (ALL), 14 cases of acute myeloblastic leukemia (AML), and 3 cases of chronic myelocytic leukemia (CML). There were 47 males and 31 females ranging from 1 to 15 years old. Bone marrows were collected. The leukemic cells were separated and cultured in the presence or absence of $10{\mu}M$ curcumin for 48 hours. MDR1 mRNA levels were determined by RT-PCR. It was found that curcumin reduced MDR1 gene expression in the cells from 33 patients (42%). Curcumin affected the MDR1 gene expression in 5 of 11 relapsed cases (45%), 10 of 26 cases of drug maintenance (38%), 7 of 18 cases of completed treatment (39%), and 11 of 23 cases of new patients (48%). The expression levels of MDR1 gene in leukemic patient cells as compared to that of KB-V1 cells were classified as low level (1-20%) in 5 of 20 cases (25%), medium level (21-60%) in 14 of 32 cases (44%), and high level (61-100%) in 14 of 20 cases (70%). In summary, curcumin decreased MDR1 mRNA level in patient leukemic cells, especially in high level of MDR1 gene groups. Thus, curcumin treatment may provide a lead for clinical treatment of leukemia patients in the future.

• Korean Journal of Clinical Laboratory Science
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• v.41 no.4
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• pp.167-172
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• 2009

Methotrexate (MTX) in one of the antineoplastic drug and it is known to effective to management of acute lymphoblastic leukemia in children, management of choriocarcinoma and related trophoblastic tumors in women, management of carcinomas of the breast, tongue, pharynx, and tests, maintenance of remission in leukemia and treatment of serve, debilitating psoriasis. Intermediate to high-dose methotrexate administration followed by leucovorin rescue is effective in treatment of carcinoma of the lung and osteogenic sarcoma. Intrathecal administration is effective in treating meningeal leukemia or lymphoma. There are FPIA (Fluorescence polarization immunoassay) and EMIT (Enzyme multiplied immunotechique) methods that measure for MTX. We evaluated the FPIA and EMIT methods. MTX were measured by Hitachi-7600 P-module using EMIT and FPIA using TDX in the sera 60 patients. The performance characteristics evaluated were, light influence, linearity, comparison with FPIA. Also, precision evaluated were three level controls through put following CLSI evaluation protocols (EP10-A). When the MTX value of $4.16{\pm}5.78{\mu}{\mu}mol/L$ (mean, SD) by the Hitachi-7600 P-module was compared with that of $4.05{\pm}5.47{\mu}{\mu}mol/L$ by FPIA, coefficients of correlation of 0.988 was obtained. The regression equation was Y (Hitachi-7600 P-module) = 0.9408 x (FPIA) + 0.1316 (r=0.9885, n=60). CVs of MTX measured by Hitachi 7600 P-module was 6.78% at $0.33{\mu}{\mu}mol/L$, 0.96% at $1.16{\mu}{\mu}mol/L$, and 0.96% at $8.04{\mu}{\mu}mol/L$. The precision was excellent in each group. The linearity was acceptable. We evaluated that MTX is light-sensitive on prolonged exposure to direct sunlight. Comparing with the FPIA using TDX, the Hitachi-7600 P-module using EMIT showed good coefficient of correlation and precision. Therefore the Hitachi-7600 P-module can replace the FPIA for quantitative analysis of MTX.

• Clinical and Experimental Pediatrics
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• v.51 no.1
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• pp.67-72
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• 2008

Purpose : In this study, we retrospectively analyzed the clinical outcomes of patients who underwent allogeneic hematopoietic stem cell transplantation (HSCT) grafted from HLA-matched parents. Methods : Seven children with acute leukemia (4 acute lymphoblastic leukemia, 3 acute myeloid leukemia) in first complete remission received allogeneic HSCT from their respective parents at the St. Marys Hospital between April, 1999 and October, 2005. The median age of patients at transplantation was 5 years (range, 1-11 years; 2 male, 5 female) and the median age of donors was 35 years (range, 30-41 years; 5 male, 2 female). We investigated the clinical outcomes such as engraftment, acute and chronic graft-versus-host disease (GVHD), transplant-related morbidity and mortality, relapse and survival. Results : Median time from transplantation to last follow-up was 69.5 months (range, 18.8-96.5 months). All patients were successfully engrafted, with a median time of 11 days (range, 10-16 days) and 26 days (range, 13-39 days) for neutrophil and platelet recovery, respectively. Grade II acute GVHD occurred in 3, and grade III acute GVHD in 1 of 7 recipients. Extensive chronic GVHD developed in 2, and limited chronic GVHD in 1 of 7 recipients. Death from transplant-related complications occurred in 1, and relapse occurred in 1 of 7 recipients. Estimated 5-year overall survival was $83{\pm}15%$. Conclusion : The clinical outcomes of recipients who underwent HSCT from HLA-matched parents were comparable to those of patients who received HSCT grafted from HLA-matched sibling donors in childhood leukemia. HLA typing of parents, as well as siblings will increase the likelihood of finding an HLA-matched family donor for patients who need HSCT.

• The Korean Journal of Nuclear Medicine
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• v.37 no.5
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• pp.308-316
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• 2003

Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.

• Clinical and Experimental Pediatrics
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• v.48 no.8
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• pp.839-845
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• 2005

Purpose : Neck masses, in pediatric population, derive from a multitude of congenital, inflammatory, or neoplastic diseases. The majority of these masses represent benign conditions. However, thorough clinical evaluation is required to rule out malignant diseases. We evaluated the causes, clinical characteristics and outcomes of children with neck masses who underwent tissue biopsy. Methods : A total of 28 medical records of children with neck mass who underwent tissue biopsy at Chungnam National University Hospital, from January 2000 to March 2004 were retrospectively analyzed. The methods of biopsy were ultrasonography guided core biopsy(CB), fine needle aspiration biopsy(FNAB) and excisional biopsy. Results : Out of 28 patients, half were boys. The most common location of the mass was the posterior cervical area(N=19, 67.9%). Laboratory findings of peripheral blood and serologic studies were nonspecific. In 25(89.3%) cases, CB or FNAB was initially performed for neck masses. Among them 10 cases(40%) were reactive hyperplasia, 8(32%) inflammatory granulation tissues, 4(16%) necrotizing lymphadenitis, and 3(12%) acute suppurative inflammations. Initially, excisional biopsy was performed for diagnosis in 3(10.7%) cases. Diagnosis of these cases was thyroglossal duct cyst, dermoid cyst and lymphoblastic lymphoma, respectively. Conclusion : Most neck masses in children were benign. CB and FNAB were safe methods for tissue sampling, without need for general anesthesia.

• Clinical and Experimental Pediatrics
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• v.45 no.8
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• pp.1000-1006
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• 2002

Purpose : In the course of treatment, patients with hematological or oncological disorders often develop pulmonary complication. The patients who develop a severe pulmonary complication have a poor outlook. The causes of pulmonary complication are either infectious or non-infectious in origin. We have analyzed the etiology and outcome of these patients admitted to the pediatric intensive care unit of Asan Medical Center. Methods : Medical records of 95 patients on Pediatric oncology service who were admitted to pediatric intensive care unit(PICU) of Asan Medical Center from Jan 1997 to May 2000 were retrospectively reviewed. Results : The mean age of the patients was 8.5 years(2 months-18 years). The underlying malignancies of these 95 patients were as following; acute lymphoblastic leukemia(31 cases), lymphoma (11 cases), acute myeloid leukemia(nine cases), brain tumor(eight cases) and other solid tumors(25 cases). Pulmonary complications included pneumonia, acute respiratory failure, pneumothorax and pleural effusion. The most common cause of pulmonary complication was infection(88%) in etiology. The overall mortality rate was 56.8%. Pulmonary complications in these patients carried high rates of mortality regardless of whether they were immune compromised(76%) or not(69%). Even without pulmonary complications, the hematological or oncological patients admitted to PICU had high mortality rates of 43%. Conclusion : Pulmonary complications are frequent finding in the hematological or oncological patients admitted to Intensive Care Unit. The main etiology of these pulmonary complications was infection, which carried a high mortality rate regardless of their immune status at the time when they were admitted to PICU.