No abstract available.

BACKGROUND: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB CD34+ cells. METHODS: CD34+ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-1 beta, interleukin-6, granulocyte macrophage-colony stimulating factor and tumor necrosis factor-alpha were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB CD34+ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule- 1, intercellular adhesion molecule- 1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. RESULTS: Several cytokines were found to have been produced by CB CD34+ cells as well as bone marrow-derived CD34+ cells. During ex vivo expansion of CB CD34+ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7- 10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0. 1% poly-L-lysine-coated wells. CONCLUSION: Stromal cells were developed during ex vivo expansion of CB CD34+ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.
Cell Adhesion ; Cytokines ; E-Selectin ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix ; Fetal Blood* ; Granulocytes ; Hematopoiesis ; Hematopoietic Stem Cells ; Interleukin-1beta ; Interleukin-3 ; Interleukin-6 ; Stem Cells ; Stromal Cells* ; Tumor Necrosis Factor-alpha ; Vimentin ; von Willebrand Factor

Due to improvements in medical care, the socioeconomic level and public health, life expectancy has dramatically increased. Thus, advances in the development of life-support systems and the control of infection have resulted in many surgical and anesthetic procedures being performed on extremely elderly patients. In contrast to younger patients, elderly patients may manifest more than one pathologic process associated with progressive degenerative changes in various organs of the aged, especially in the heart, brain, and kidney. Since both progressive degenerative change occurring in the elderly population and the cumulative incidence of disease in that population result in death intraoperatively or during the immediate postoperative period, the anesthesiologist must be particularly alert to the possibility of anesthetic risks in the elderly. The elderly patient is more likely to have hypertension, congestive heart failure, cardiac dysrhythmias, chronic pulmonary disease, and diabetes. Preoperative evaluation and treatment of those conditions must be extensively reviewed prior to the induction of anesthesia. To evaluate geriatric anesthetic experiences, 539 cases of patients aged over 60 years of 4,266 anesthetic cases admitted to P.M.C. from January to December, 1986 were analyzed according to age, sex, physical status, anesthetic technique an6 agents, surgical department, preoperative chest X-ray findings, preoperative E.C.G findings, and postoperative complications. The results are as follows. 1) Of 4,266 anesthetic patients 539(12.6%) were over 60 3ears of age and 322(59.7%) were males and 217(40.3%) females. 2) In the classification of physical status, the most common evidence was class 2 in 303 cases. Emergency surgery comprised 27.1%. 3) The anesthesia technique employed was usually general anesthesia and this suggested that balanced anesthesia used with narcotics offers several advantages to geriatric patients. 4) In the surgical department, 310 cases(57.5%) were for general surgery, 75 cases(13,9%), orthopedic surgery; 57 cases(10.6%), urology; and 49 cases(9.1%), neurosurgery, respectively. Cancer was present in 198 cases(36.7%), 5) Preoperative chest X-ray findings: The most common finding was pulmonary tuberculosis in 44 cases(8.2%). Pneumonia, pulmonary emphysema, and so forth were also observed. 6) Preoperative E.C.G findings: The most common findings was myocardial ischemia in 48 cases(8.9%). Also myocardial infarction observed in 8 cases(1.5%) 7) Postoperative complications were as follows: The most common incidence was wourid infection in 29 cases(5.4%) followed by pneumonia. There were a number of miscellaneous complications. but postoperatively, they did not present any significant problems. 8) The overall mortality rate was 3.5%(19 cases). The difference in the mortality rate related to age was not statistically significant(p>0.1), but the mortality rate related to physical status was statistically significant(p<0.001). 9) Optimizing a patient's preoperative condition by the anesthesiologist, consultants, and other physicians was assumed to reduce perioperative morbidity and mortality.
Aged ; Anesthesia* ; Anesthesia, General ; Arrhythmias, Cardiac ; Balanced Anesthesia ; Brain ; Classification ; Consultants ; Emergencies ; Female ; Heart ; Heart Failure ; Humans ; Hypertension ; Incidence ; Kidney ; Life Expectancy ; Lung Diseases ; Male ; Mortality ; Myocardial Infarction ; Myocardial Ischemia ; Narcotics ; Neurosurgery ; Orthopedics ; Pneumonia ; Postoperative Complications ; Postoperative Period ; Public Health ; Pulmonary Emphysema ; Thorax ; Tuberculosis, Pulmonary ; Urology

PURPOSE: Thrombopoietin (TPO) has been currently used for ex vivo expansion of hematopoietic stem cells. Previously, we have reported that TPO induces apoptosis during ex vivo expansion of cord blood CD34 cells. In this study, we have investigated the stem cell factor (SCF) to determine whether it affects the TPO-induced apoptosis. METHPDS: CD34+ cells, purified from four separate human cord bloods by magnetic bead selection, were expanded in Iscoves modified Dulbeccos medium with several cytokines. Apoptosis has been confirmed by several ways; 7-amino-actinomycin D (7-AAD) or annexin V staining, and morphologic analysis with light and electron microscopic examinations. And cell maturation was also analysed with DNA staining. RESULTS: Apoptosis was reached peak (67.8+/-5.24% of total cells) at day 9 in the cultures with TPO alone. Morphological examination of Giemsa-stained cytospin preparations revealed many immature cells with cytoplasmic blebbing and eccentric nuclei. However, multilobed nuclei were rarely observed while bilobed nuclei were frequently observed. These results suggest that premature senescence occurs in the megakaryocytic cells before full polyploidization. SCF was affective for the TPO-induced apoptosis; the peak apoptotic fraction was significantly decreased to 37.2+/-3.31% at day 9. In the cultures with SCF, the fractions of CD41+ cells were also decreased in parallel with decrease in apoptosis. Notwithstanding reduction of differentiation into megakaryocytic lineage, polyploidization(> 6N) in CD41 cell faction was significantly increased in the cultures with SCF at day 9 and day 11, respectively (P<0.05). CONCLUSION: These results suggest that SCF inhibits premature senescence of the differentiating cells of megakaryocytic lineage during ex vivo expansion using TPO.
Aging* ; Annexin A5 ; Apoptosis ; Blister ; Cytokines ; Cytoplasm ; DNA ; Fetal Blood* ; Hematopoietic Stem Cells ; Humans* ; Megakaryocytes* ; Polyploidy ; Stem Cell Factor* ; Stem Cells* ; Thrombopoietin*

PURPOSE: We aimed to study the feasibility of arsenic trioxide as a treatment of neuroblastoma which has the ability to differentiate into nonmalignant cells like acute promyelocytic leukemia. METHODS: To determine the effects of arsenic trioxide in various concentrations and exposure time on the survivial of neuroblastoma cell lines, SH-SY5Y and SK-N-AS cells were cultured in RPMI 1640 media with 1 to 20muM concentration of arsenic trioxide. Apoptosis was measured with flow cytometry by staining with 7-aminoactinomycin D. Cell cycle was assessed by monitoring the DNA contents by flow cytometry. Arsenic trioxide induced cell morphologic changes were also observed with May-Grunwald-Giemsa stain under a light microscope. RESULTS: Arsenic trioxide induced apoptosis in SH-SY5Y cells earlier in the same concentration and to a more severe degree with the same exposure time than in HL-60 cells. The apoptosis induced by arsenic trioxide was steeply increased to 79.3 10.1% at 24 hours and then maintained a plateau on 20muM concentration, while increasing steadily to 40.2 6.5% until 72 hours on 5muM concentration. The proliferating cell proportion in S/G2/M phase was decreased with arsenic trioxide concentration and with exposure time in both SH-SY5Y and HL-60 cells, especially more so with the SH-SY5Y cells. The cellularity was decreased and more apoptotic cells could be observed in the arsenic trioxide treatment group than in untreated control group. CONCLUSION: As in acute promyelocytic leukemic cells, arsenic trioxide induced apoptosis and cell cycle arrest of proliferating phase in neuroblastoma cells.
Apoptosis ; Arsenic* ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Line ; DNA ; Flow Cytometry ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; Neuroblastoma*

About 15 to 20% of ovarian tumors are of germ cell origin, and about 95% of these are cystic teratoma. when the teratoma is composed either entirely or predominantly of thyroid tissue, it called struma ovarii. Struma ovarii represents about 2.7% of cystic teratoma. The incidence of malignant struma ovarii is probably much less than 5% of all struma ovarii. We have observed a case of malignant struma ovarii originated from the left ovary of a 46-year-old woman, and present this case with a brief review of concerned literatures.
Female ; Germ Cells ; Humans ; Incidence ; Middle Aged ; Ovary ; Struma Ovarii* ; Teratoma ; Thyroid Gland

Bcr-abl antisense oligodeoxynucleotides (AS-ODNs) have provided evidence of an antileukemia effect when tested in vitro against Philadelphia-positive cells. In order to investigate the efficacy of AS-ODNs as purging agents in chronic myeloid leukemia (CML) patients, K562 cells, a human CML cell line, were treated in vitro with various types of AS-ODNs and interferon-alpha. Cells were treated in vitro for 0 and 36 hr with 40 microgram/mL of AS-ODNs, respectively, and incubated at 37 degrees C for 36 hr. Cytotoxic effects were measured by counting the number of viable cells as well as by MTT test. Clonogenic activities were evaluated by methylcellulose culture for 2 weeks. The effects of purging agents on the rearrangement of bcrabl gene were evaluated by RT-PCR. AS-ODNs inhibited the proliferation of K562 cells with time in cell count assay and MTT test. AS-ODNs were superior to INF-alpha in inhibiting clonogenic activity (recovery rate; 26.3% vs 64.0%). After incubation with bcr-abl AS-ODNs primers and mRNA isolated from K562 cells, positive bands were abolished, especially of b3a2 type and phosphorothioate type. Our results suggest that AS-ODNs mediated purging may be one of the efficient methods and that autograft may be an alternative treatment for allograft in high-risk group patients of CML if they do not have a stem cell donor.
Bone Marrow Purging* ; Colony-Forming Units Assay ; Fusion Proteins, bcr-abl/genetics ; Fusion Proteins, bcr-abl/metabolism ; Hematopoietic Stem Cells/physiology* ; Human ; Leukemia, Myeloid, Chronic/therapy ; Neuroblastoma/therapy* ; Oligonucleotides, Antisense/metabolism* ; Oligonucleotides, Antisense/therapeutic use ; Transplantation, Autologous* ; Tumor Cells, Cultured

PURPOSE: Bronchopulmonary dysplasia (BPD) is characterized by arrested vascular and alveolar growth in the premature lung. Considering the consequences of arrested lung growth, the idea of administering bone marrow cells to enhance the inborn repair mechanism is promising as this may reduce the morbidity and mortality of BPD. We followed enhanced green fluorescent protein (EGFP)-labeled bone marrow cells (BMC) injected intraperitoneally into non-EGFP mice in order to determine their fate after transplantation. METHODS: An angiogenesis inhibitor, SU1498, was injected subcutaneously on day 3 in non-EGFP C57BL/6 newborn mice to create a model of arrested alveolar development. On the following day, 1x10(6) BMCs isolated from major histocompatibility complex (MHC)- matched syngenic EGFP mice were injected intraperitoneally to non-EGFP BPD mice. Morphometric analysis, immunostaining, and confocal microscopy were performed to determine the fate of EGFP-positive stem cells in the injured lung. RESULTS: SU1498 injection reduced alveolar surface area and mean alveolar volume in newborn mice. BMC injection resulted in recovery of lung structure comparable to controls. EGFP-positive BMCs were identified in the lungs of the recipient mice after intraperitoneal injection. The injected EGFP cells were co-stained with endothelial and epithelial cells of the developing lung as determined by confocal microscopy. CONCLUSION: Our results illustrated that EGFP-positive BMCs engrafted and trans- differentiated into epithelial and endothelial cells after intraperitoneal injection in a mouse model of arrested alveolar development.
Animals ; Bone Marrow ; Bone Marrow Cells ; Bronchopulmonary Dysplasia ; Cinnamates ; Endothelial Cells ; Epithelial Cells ; Green Fluorescent Proteins ; Humans ; Infant, Newborn ; Injections, Intraperitoneal ; Lung ; Major Histocompatibility Complex ; Mice ; Microscopy, Confocal ; Stem Cells

Chronic graft versus host disease (GVHD) is a frequent complication after allogeneic hematopoietic stem cell transplantation (HSCT), but simultaneous small bowel obstruction is rare. Here, we report a child with acute myeloid leukemia who received an allogeneic HSCT from an unrelated matched donor. After HSCT, the patient developed severe chronic GVHD involving the small intestine, leading to obstruction of the terminal ileum. Small bowel resection was performed, and the symptoms improved without severe complications. Bowel obstruction should be considered as a possible complication of chronic GVHD; surgery may be a valuable corrective measure.
Child ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Ileum ; Intestinal Obstruction ; Intestine, Small ; Leukemia, Myeloid, Acute ; Tissue Donors ; Transplants

No abstract available.
Brain Neoplasms* ; Brain* ; Child* ; Humans ; Infant*