Clinical effects of KRN8601 (recombinant human granulocyte colony-stimulating factor: rhG-CSF) were studied in 26 patients with chronic neutropenia including 4 Kostmann's disease, 1 Shwachman's syndrome, 1 Lonsdale's syndrome, 1 glycogen storage disease Ib-associated, 6 chronic benign, 5 chronic hypoplastic, 2 cyclic, 4 autoimmune and 2 miscellaneous neutropenia. The patients were given rhG-CSF intravenously at doses of 20-540 μg/m² or subcutaneously at doses 20-400 μg/m², over the periods of 2-32 weeks. Increases in neutrophil conunts occurred after rhG-CSF administration in 23 of the 26 patients. Patients with Kostmann's disease, Shwachman's syndrome and chronic hypoplastic neutropenia responded poorly compared to patients with other types of neutropenia. There were no serious side effects which caused interruption of the study. These results indicated a beneficial effect of KRN8601 in various types of chronic neutropenia.

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Glanzmann's thrombasthenia (GT) is an autosomal recessive bleeding disorder due to a deficiency or abnormality of glycoproteins (GPs) IIb and IIIa, but its genetic basis remains to be determined. We analyzed the genes for GPIIb and 3'GPIIIa in 3 patients with GT and in 7 control subjects by Southern blot. No large deletions or insertions were detected in these genes in any patient with GT. Furthermore, the GPIIb and GPIIIa mRNAs derived from the platelets of patients with GT could be amplified using the reverse transcriptase-polymerase chain reaction (RT-PCR). This finding indicates that the mRNAs encoding for GPIIb and GPIIIa are present in the patients' platelets. Thus, the GPIIb and GPIIIa genes of the patients with GT are considered to be normally transcribed. The molecular defects of the GPIIb and/or GPIIIa genes in our patients remain to be clarified.

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This is a report of Fechtner syndrome in two Japanese families. Six members of family I and three members of family II were studied. All but one had macrothrombocytopenia and leukocyte inclusion bodies, four had deafness, four had persistent proteinuria and none had cataracts. Under a diagnosis of ITP, two of them had splenectomy which resulted in no response. History reveolod, other family members with deafness and/or nephritis were confirmed in both families. Ultrastructural studies of leukocytes showed oval inclutson bodies with unclear borders containing many fine ribosome like granules and randomly scattered filaments. Ultrastructural studies of macrothrombocytes were unremarkable except for a well-developed opencanalicular system. More than half of megakaryocytes had uneven basophilic speckles in the cytoplasm, which were positive for Unna-Pappenheim staining. Ultrastructuraly, widening of demarkating systems and remaining ribosomes were noted in the cytoplasma of mature megakaryocytes.

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The clinical safety and efficacy of transfusion of red cell concentrates stored in MAP solution (MAP-CRC) containing mannitol, adenine, glucose, phosphate and citrate, into 39 anemic patients were evaluated. In 23 patients, infusion of MAP-CRC was alternated with infusion of ordinary CRC as a control. The MAP-CRC and CRC used in this study were stored at 4°C for an average of 38.2±2.6 days (n=52) and 18.1±2.2 days (n=26), respectively. Red cell recovery was 77.5% for MAP-CRC and 82.5% for CRC, based on calculation of the increase in hemoglobin level one day after transfusion. There were no differences between patients transfused with MAP-CRC and those transfused with CRC in clinical findings or biochemical data. No major side-effects other than pyrexia associated with the underlying infections were seen in patients transfused with MAP-CRC. MAP-CRC stored up to 42 days is apparently as safe and effective as stored CRC. This new additive solution may therefore be useful for the future expansion of the indications for autologous blood transfusion by facilitating the collection and storage of more blood in the liquid state for a longer period, and may also be useful in obtaining more plasma from whole blood as source plasma.

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The efficacy of traditional Chinese medicine (Kampo medicine) was examined in 40 patients with steroid-unresponsive idiopathic thrombocytopenic purpura (ITP). All patients were, at first, given Hochuekihi-to, and when they were refractory, therapy was switched to Sairei-to, Ninjinyoei-to, and Kamikihi-to. The efficacy rate was 20.0% by Hochuekihi-to, 20.7% by Sairei-to, 5.5% by Ninjinyoei-to and 6.7% by Kamikihi-to, respectively. Thus, overall responders were 12 in 40 patients with an effective rate of 30.0%. The mechanism of action of traditional Chinese medicine is discussed in relation to the production of various kinds of cytokines.

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Affected erythrocytes in patients with paroxysmal nocturnal hemoglobinuria (PNH) have been detected as complement-sensitive cells by the complement sensitivity assay. Decay-accelerating factor (DAF), a molecular mass of 70 kDa complement-regulatory membrane glycoprotein, has been reported to be deficient on affected PNH blood cells. In the present study, DAF on erythrocytes from 12 patients with PNH were stained by an immunocytochemical method and the ratio of DAF⁺ erythrocytes was compared with their laboratory data concerning hemolysis. almost all normal human erythrocytes stained positively for DAF. In contrast, various percentages of DAF⁺ erythrocytes were found in the patients with PNH. The percentages of DAF⁺ erythrocytes correlated positively with the total amounts of DAF on erythrocytes measured by an enzyme-linked immunosorbent assay (ELISA), negatively with % hemolysis in Ham's test and in sucrose hemolysis assay and with percentages of PNH type III (PNH-III) erythrocytes. In some patients with PNH, subpopulations of erythrocytes weakly-positive for DAF were demonstrated in addition to DAF^- erythrocytes. The immunocytochemical method for staining DAF on erythrocytes developed in the present study is useful for the detection of affected PNH erythrocytes and applicable to further studies on membrane defects in PNH.

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It is generally considered that abnormality of the erythrocyte membrane skeleton co elliptocytes. There are, however, few reports of β spectrin variants. We found a new variant of β spectrin in a child and her mother. This report is the first case of abnormality of β spectrin in Japan. The propositus was an 8 month-old girl who was first examined by us in 1988. On laboratory findings, she showed anemia, increased reticulocyte count and decreased haptoglobin concentration. Both peripheral blood smears of patient and her mother showed typical elliptocytosis and they were diagnosed as hereditary elliptocytosis. SDS-PAGE patterns of the red cell membranes of the propositus and her mother were characterized by the presence of an abnormal component migrating immediatetely below the spectrin chains. We confirmed that the abnormal spectrin appeared clearly at the expense of normal β chain. The abnormal spectrin (M. W. 216,000d) makes up 16% of the total β chain. The inheritance of our case was autosomal dominant. The present case is considered as a new spectrin variant.

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An 8 year-old boy, who had been healthy since his birth, presented fever, ecchymosis and nasal bleeding. His peripheral blood picture showed pancytopenia and bone marrow showed erythroblastopenia, but these findings recovered spontaneously in two weeks. The diagnosis of human parvovirus B19 (HPVB19) infection was made by the presence of anti HPVB19 IgM antibodies in the sera. In order to investigate the mechanisms of erythroblastopenia and pancytopenia, we performed in vitro hematopoietic cell culture in both acute phase and the convalescent phase (3 months after the onset). In the acute phase, the formation of CFU-E colony was inhibited but that of BFU-E, CFU-G, GEMM colony was not inhibited. Unexpectedly, HPVB19 DNA was detected in CFU-GEMM colony cells with the PCR technique. It is suggested that HPVB19 infected not only to erythroid progenitor cells but also to other progenitors.

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Myelodysplastic syndrome (MDS) combined with monoclonal gammopathy or multiple myeloma has rarely been reported. In this article, two siblings, a brother and his sister who showed simultaneous occurrence of MDS and monoclonal gammopathy are reported. The first case, a 73-year-old male, was admitted to our hospital in November, 1987. Analysis of peripheral blood revealed pancytopenia without blast cells. Bone marrow was hypocellular with 14.9% of myeloblasts and 2.8% of plasma cells characterized by 2 to 4 nuclei. Serum IgA level was 635 mg/dl and serum immunoelectrophoresis revealed a monoclonal IgAλ band. The second case, a 70-year-old female, younger sister of the first case, was admitted to our hospital in January, 1988. Bone marrow was normocellular with 23% of peroxidasenegative myeloblasts and 12.8% of atypical plasma cells. Serum IgG level was 1,901 mg/dl with monoclonal IgGκ band. Hematological findings have remained unchanged for 12 months. The first case was regarded as hypoplastic MDS with monoclonal gammopathy and the second case was MDS with smoldering myeloma. These cases were very similar with in respect to age, time of onset, clinical course, hematological findings and especially, association with M-protein. There are no reports concerning the familial incidence of MDS with M-protein. These findings supported the hypothesis that an initial event selects a clone of stem cells which retain the capability to differentiate into mature myeloid and lymphoid cells, in these cases B-cells.

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Myelodysplastic syndromes that occurred in two young brothers are reported. A 19-year-old man was admitted to Kobe City General Hospital in May 1990 because of fever and nasal bleeding. On admission his hemoglobin was 5.5 g/dl, platelet count 1.5×10⁴/μl and white cell count 1,700/μl with 18% neutrophils and 80% lymphocytes. Bone marrow aspirate showed dysplastic features of trilineage blood cells with 4.8% myeloblasts. A diagnosis of refractory anemia was made. His younger brother, a 17-year-old man was examined in May 1990 because of increasing fatigability of 2 years' duration. His hemoglobin was 8.7 g/dl, platelet count 2.1×10⁴/μl and white cell count 2,800/μl. Bone marrow aspirate revealed morphological abnormalities in three lineages with 5.2% myeloblasts. He was diagnosed as having refractory anemia with excess of blasts. Their parent are consanguineous. The onset at a young age, reduced CD4 lymphocytes and similarity of dyshematopoietic findings suggests the presence of common genetic disorder in the pluripotent hematopoietic stem cells.

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Myelodysplastic syndrome (refractory anemia with excess of blasts; RAEB) with marked basophilia and eosinophilia is described. An 82-year-old male was admitted to our hospital because of severe normocytic normochromic anemia (Hb 5.6 g/dl). The white cell count was 9,200/μl with marked basophilia (34.5%) and eosinophilia (19.5%). The bone marrow aspiration also revealed both basophilia and eosinophilia, with blast contents of 9%. Diagnosis of RAEB was established. Although the treatment with red cell transfusion and ubenimex (Bastatin) was started, anemia was not improved. A karyotype of the bone marrow cells from this patient showed 47, XY, +8, i(17q), which has been observed as additional chromosomal abnormalities in blastic crisis of chronic myelogenous leukemia. The diagnosis of CML was not compatible with this case, because Ph¹ chromosome and bcr gene rearrangement were negative. It is concluded that eosinophilia and basophilia might be derived from clonal abnormalities associated with MDS.

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A 37-year-old woman was admitted to our hospital because of anemia in the 33rd week of pregnancy. On admission, the hemoglobin was 7.6 g/dl, platelets 508,000/μl and WBC 8,300/μl with 4% blast cells. Bone marrow aspirate demonstrated 50% erythroblasts of nucleated cells, which had prominent megaloid and polynucleic changes, and 20% myeloblast cells. We diagnosed her as having erythroleukemia. Receiving packed-red-cell transfusions, she had a cesarean section, and gave birth to a female infant (BW 2,175 g) in the 34th week of pregnancy. At the same time, she had a total hysterectomy with left adnexectomy. The postoperative course was favorable. She had a combination chemotherapy of BHAC-DMP, resulting in a partial remission. After the second induction chemotherapy with subcutaneous use of G-CSF and BHAC-DMP, complete remission was obtained, which lasted for almost 17 months. The child is growing well without any hematological disorder.

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A 74-year-old Japanese male was admitted because of anemia. Hepatosplenomegaly, lymphoadenopathy, and purpura were not found. The laboratory data on admission revealed that the white-cell count was 9,400/μl, the hemoglobin 11.1 g/dl, and the platelet count 17,000/μl. Platelet-associated IgG was 794.2 ng/10⁷ cells. The patient was diagnosed as having autoimmune thrombocytopenic purpura (ATP) at this time. He was treated with prednisolone, but his thrombocytopenia not improve. In addition to prednisolone, azathioprine was given to him. During the course of treatment, leukocytosis gradually appeared and the white-cell count reached more than 30,000/μl with over 70% lymphocytes. A bone marrow aspiration revealed 70% of small lymphocytes, and surface marker analysis showed that CD19 and HLA-DR were positive on these lymphocytes. Southern blotting analysis demonstrated rearrangements of JH and JK. He was finally diagnosed as B-CLL complicated by ATP. One month after the azathioprine administration, the platelet count increased more than 30,000/μl and the white-cell count decreased less than 10,000/μl. About 2% of patients with CLL are known to be complicated by ATP. To our knowledge, the present case is the first case of B-CLL complicated by ATP in Japan.

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A 70-year-old man was admitted to our hospital on March 9, 1989 because of fever, superficial generalized lymphadenopathy, upper abdominal mass and right pleural effusion. The diagnosis of non-Hodgkin's lymphoma (follicular medium sized cell type, B cell) was made by a biopsy of the neck lymph node. Peripheral blood mononuclear cells were obtained from the patient by cytopheresis. The cells were cultured for 8 days with interleukin-2 (IL-2) to generate Lymphokine-activated killer (LAK) cells. The patient received a total of 7.7×10⁹ LAK cells intravenously over a period of 3 weeks. He also received continuous intravenous infusion of IL-2 for 17 days, starting 2 days before the first infusion of LAK cells. After this therapy, although his superficial generalized lymphadenopathy disappeared or decreased in size, the size of the upper abdominal mass did not decrease. Therefore, it is suggested that adoptive immunotherapy is a beneficial treatments for B cell lymphoma. However, LAK cells should be generated in much larger quantities for a more successful therapeutic result.

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This paper reports a rare case of Hodgkin's disease with Sjögren syndrome in the course of Behçet's disease. A 43-year-old man developed arthralgia of bilateral knees, ankles, elbows and wrists in May, 1988. He had hazy vision and was diagnosed as having iridocyclitis and chorioretinitis in February, 1989. Gingival ulcer, penile ulcer, erythema nodosum on the right lower leg and superficial thrombophlebitis on the bilateral arms appeared in June, 1989. Therefore, he was diagnosed as Behçet's disease. He responded well to prednisolone. In November, 1989, he developed fever with positive CRP and elevated alkaline phosphatase. Multiple mass lesions in the liver and spleen with retrocrural lymphadenopathy were noticed on the abdominal CT and echogram. A cervical lymph node biopsy revealed Hodgkin's disease of the mixed cellularity type. At the same time, the patient had dry eyes and a dry mouth. Salivary gland biopsy revealed chronic sialoadenitis with lymphocytic infiltration compatible with Sjögren syndrome. The patient responded well to ABVD regimen. He is still free of disease as of May, 1991.

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A 34-year-old man was admitted in May, 1989 because of severe left upper abdominal pain, which was caused by advanced splenomegaly. On initial examinations, peripheral blood showed leukocytosis (13,400/ul) including 60% hairy cell which also infiltrated in bone marrow (64%). The patient was diagnosed as having hairy cell leukemia (Japanese type) of B-cell lineage. Splenectomy was performed as an initial treatment. The effect of splenectomy was only palliative and transient leukocytosis progressed thereafter. α-interferon and bestrabucil (KM 2210) were then adopted for 4 months respectively. The effects were, however, unsatisfactory. Subsequently the patient was treated successfully with 2'-deoxycoformycin (DCF). Complete remission was attained following 12 injections of 7.5 mg/body of DCF during 5 months and durable remission persists for more than 6 months without maintenance therapy. The side effect was minimun.

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A 38-year-old male was admitted in January 1984 due to lymphadenopathies with hyperimmunoglobulinemia with a serum IgG level of 2,872mg/dl. Following this, he was observed as an outpatient in regard to lymphadenopathies of unknown origin. In 1989, after the fourth lymph node biopsy he was diagnosed as having idiopathic plasmacytic lymphadenopathy with polyclonal hyperimmunoglobulinemia. At that time his serum IgG level was 8,090mg/dl. The elevated serum interleukin-6 (IL-6) level, up to 21.1pg/ml, was particularly interesting, because IL-6 is involved in the oncogenesis of plasmacytoma/myeloma. The patient also had thrombocytosis, hematuria, and a serum increased level of C reactive protein which seemed to be related to the effects of IL-6 i.e. thrombopoiesis, induction of the proliferation of mesenchymal cells, and induction of the production of acute phase proteins by hepatocytes, respectively. Even though he displayed no outward symptoms before and after treatment with prednisolone and melphalan, elevated immunoglobulin levels were still present.

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A 57-year-old woman visited to our hospital complaining of paresthesia in the right leg. She had no abnormal physical findings. However, the peripheral blood examination demonstrated 7% basophilia with 8000/μl WBC count and decreased neutrophil alkaline phosphatase activity (score 37, rate 19%). She was diagnosed as Ph¹ chromosome positive CML in early phase by the chromosomal analysis of bone marrow cells. She received subcutaneous injection of natural interferon-alpha at a dosage of 600×10⁴ IU daily from March 10, 1987. The dosage and administration interval were gradually reduced and prolonged. Since November 1988, weekly injections of 300×10⁴ IU has been administered as maintenance therapy. Cytogenetic improvement was seen at 4 months after the start of IFN. Disappearance of Ph¹ chromosome positive cells was observed on December 11, 1987. It was suggested that the administration of IFN from the early chronic phase played an important role in the control of the disease.

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Two patients with essential thrombocythemia were successfully treated by administering native α-interferon (α-IFN). One patient was a 38-year-old man in whom thrombocytosis was found accidentally. His platelet count on admission was 880,000/μl and megakaryocytes increased. Three million units of α-IFN was administered subcutaneously everyday, and the platelet count decreased gradually to about 500,000/μl within 2 weeks. The other patient was a 66-year-old woman who visited our hospital complaining of tenderness and swelling of the fingertips. Her platelet count was 1,610,000/μl, and megakaryocytes increased and showed abnormal morphology. Six million units of α-IFN was administered subcutaneously every other day. The tenderness and swelling of the fingertips disappeared soon after the beginning of α-IFN administration. The platelet count decreased to about 500,000/μl within 10 days, but she developed itching of the skin over the entire body. Therefore, α-IFN treatment was discontinued. It was suggested that α-IFN suppresses not only the maturation and proliferation of the progenitors of megakaryocytes but also the production of platelets from megakaryocytes. Administration of α-IFN should be considered in treating patients with essential thrombocythemia, because effects appear soon and α-IFN does not induce a second malignancy.

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We report a 9-year-old girl who had massive bone marrow infiltration of tumor cells at the onset of alveolar rhabdomyosarcoma. She was admitted to a surgical hospital because of abdominal pain and tumor in the buttock. Computerized tomographic scans of the pelvis revealed an abnormal mass. She was referred to our department. Hematological examination showed pancytopenia. Blood chemistry revealed hypercalcemia and hyperuricemia. Bone marrow was occupied with 100% tumor cells. Chromosome analysis of tumor cells in bone marrow revealed a specific translocaiton, t(2;13)(q37;q14). This finding enabled the diagnosis of a disseminated alveolar rhabdomyosarcoma to be established. She was treated with vincristine, cyclophosphamide and epirubisin, achieved a complete remission, but died of relapse 3 months after diagnosis.

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Forty five year old male suffering from relapsed acute promyelocytic leukemia (APL) was treated with all-trans retinoic acid (ATRA) and attained second complete remission (CR) without bone marrow hypoplasia. He was diagnosed as having APL in September 1989. The DCMP-85 regimen first induced CR in October, however the disease relapsed in September 1990. The DCMP-85 and the MEC (MIT, ETOP, Ara-C) regimens were applied for re-induction without success. Then, 45mg/m²/day ATRA was given orally from December 28, 1990. Laboratory data before ATRA treatment were as follows; 35.4% leukemic cells in the bone marrow, Hb 11.0g/dl, Plt 130,000/μl, WBC 5,100/μl without leukemic cells, and no DIC was detected. During the treatment, his bone marrow was examined frequently. The bone marrow series showed no hypoplasia at any time and gradual reduction of leukemic cells with proliferation of mature granulocytes. CR was attained on January 21, 1991. DIC did not develop. Cytogenetic anomalies including t(14;17;15)(q24;q11.2;q22) reduced from 29/30 cells at relapse to 4/30 cells at the time of CR. Dryness of mouth and lips, irritation around eyes and the elevations of GOT, GPT and triglyceride level were seen as the side effects of ATRA, however they were tolerable.

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We described two pediatric patients with acute promyelocytic leukemia (APL) who were successfully induced into complete remission with all-trans retinoic acid (ATRA, 45 mg/m² per day) after failing on conventional chemotherapy. Initial response was observed as correction of DIC within a week of treatment. Hematologically, initial increase of maturing leukocytes reached a peak peripheral WBC count on the 16th and 20th day, respectively. However, these seemingly differentiated leukocytes retained Auer body and dysplastic features and there was no concomitant recovery of erythroid and megakaryocytic lineages at this point. A sudden drop of leukocyte counts after this peak made a brief period of leukopenia before the complete remission was finally attained morphologically in 4∼5 weeks. Thus, remission of APL by ATRA therapy consisted of a two-phase course. In one patient, we observed an increase of histiocytes phagocytizing leukocytes in the marrow during the recovery from leukopenia. It is, therefore, postulated that the two-phase course of recovery may reflect the differentiation of leukemic cells by ATRA and subsequent clearance of senescent cells by the reticuloendothelial system followed by regeneration and differentiation of residual normal hematopoietic stem cells.

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A 64 year-old female patient of idiopathic myelofibrosis (IMF), who had had rapidly progressing massive splenomegaly and severe pancytopenia refractory to blood transfusion, was treated with PSL 0.6 mg/kg/day for a month, being significantly improved not only symptomatically and hematologically but also in bone marrow hematopoiesis. Although the effectiveness of PSL to restoration of bone marrow hematopoiesis has been almost unknown, long term oral PSL should be tried for a certain phase of IMF.

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