In this paper titled "My Kumamoto Life of 19 Years; The Travel for Times", the memorial lecture on my retirement from Kumamoto National University Corporation Integrated Medical and Pharmaceutical Sciences, Department of Biomedical Informatics (Chairman) is summarized. As they say "Time flies", time extends from seconds to years. The lecture includes a summary of my short term research and long term studies, such as age-dependant and gene-related changes in ageing over 5 or more years in the healthy elderly. Short-term study mostly involved of newly evaluated assay methods for important substances such as the second level in the cell life span in the variation of lipid metabolite of cardiovascular diseases based on atherosclerosis, Alzheimer disease, and their evaluation by homogeneous assay of HDL-C, LDL-C, enzymatic assay for choline relating metabolites, and lipoperoxide as the results of free radical reactions. The intermediate-term studies were mainly on the development of total laboratory automation (TLA) for the management of the laboratory of the university hospital. The hospital has various degrees of sophistication in its laboratory services. Technicians were allowed to transport specimens immediately by using an air-shooter system after drawing blood, from the emergency room to the central laboratory. Routine specimens could be measured within 30 min and the results could be automatically sent to the physician's office. It greatly minimized reporting errors, decreased the exposure to biohazards, reduced labor expense, improved operation efficiency, and shortened turnaround time. Moreover, for the outpatients and emergency laboratories, we constructed a robotic measuring system which was assembled into a sequential method for the analysis of chemistry, hematology and urinalysis specimens by using a polyarticular robot. The robot arm extends to a bar-coded tube, picking up and placing test tubes on a turn table of autoanalyzers for analysis without manpower. This is the first known effective unmanned procedure for assay in the world of laboratory medicine. Also, our research on pathological informatics was begun. Our laboratory had 7 themes; the study of the reference intervals in the elderly as one of strategy of standardization on laboratory data (Drs. Uji Y, Sugiuchi H), the study of the activation mechanism of ribosomal proteins by the functions of blood cells (Drs. Shibuya Y, Senba U), the evaluation of diagnostic methods in latent ailments by gene analysis (Dr. Ando Y), the evaluation of a highly sensitive method for disseminated intravascular coagulopathy (Okajima K), the study of the neogenesis of blood vessels by physical invasion (Uchiba M), the study of the deposition mechanism of amyloid proteins and evaluation of the diagnostic methods in the autonomic system(Drs. Ando Y, Nakamura M), and the study of the function of blood stem cells in blood transfusion services (Drs. Yamaguchi K, Yonemura M, Tsunemi M). Finally, my long term work also included research into the early diagnosis and prediction of latent ailments and the variation of serum proteins levels in the healthy aged. The conclusion was that the reference value of healthy populations and individuals (intra-personal) who had no combined ailments, in follow-up for 5 years, categorized by age, sex, and social conditions, gave a narrower range of variation than did large mixed populations (inter-personal), in laboratory tests and activity of daily living (ADL). Concerning the early diagnosis and prediction studies for the latent ailments in the aged, variations of serum protein levels in the healthy aged were classified into 3 types: serum proteins levels increased with advancing age (alphaAT, mainly acute phase reactant proteins), those that decreased with advancing age (albumin mainly transporting proteins) and proteins with no significant variation. The higher increase of the alpha1AT/beta2 III ratio in the healthy aged over 60 years old is suspected to create symptoms in the near future. The papers presented concerning ageing studies were presented at the APCCC Symposium (India, 2002) and the IFCC Symposium (Kyoto, 2002), and the TLA study was also presented at the Symposium of XX World Congress of Pathology and Laboratory Medicine (San Paulo Brazil 1999).

A 51-year-old woman with severe aplastic anemia underwent allogeneic peripheral blood stem cell transplantation using blood stem cells from an HLA-identical sibling. Adenovirus type 11 hemorrhagic cystitis developed and progressed to nephritis and hemophagocytic syndrome. Oral ribavirin was effective not only for the hemorrhagic cystitis and nephritis but also for the hemophagocytic syndrome. Since therapeutic strategies for adenovirus infection after hematopoietic stem cell transplantation have not been established, we present our case and discuss the therapeutic approach.

A 4-year-old boy with Down syndrome (DS) was diagnosed as having acute monoblastic leukemia (AML-M5a). Leukemic cells were CD33+, CD56+ and CD4+, with t(9;11) on cytogenetic analysis and MLL gene rearrangement. After 2 courses of induction therapy using an AML 99-Down protocol failed to obtain complete remission, the patient received cord blood transplantation from an HLA-matched donor (CBT) following a conditioning regimen comprising total body irradiation and cyclophosphamide. Only cyclosporin A was used for graft-versus-host disease prophylaxis. Stem cell transplantation may not be indicated for AML patient with DS in first remission, who display a high rate of life-threatening and fatal toxicity on therapy. This patient remained well controlled in complete remission for 4 years, representing a rare case of DS with chemotherapy-resistant AML successfully treated with a CBT.

We describe 2 cases of conventional therapy-resistant multiple myeloma (MM) that responded to bortezomib and dexamethasone therapy. Case 1: A 62-year-old woman with MM (IgG, kappa-type, stage IIIA) resistant to DMVM-IFN (dexamethasone, ranimustine, vincristine, melphalan, interferon-a), VAD (vincristine, doxorubicin, dexamethasone), high-dose melphalan with autologous peripheral blood stem cell transplantation (PBSCT) and thalidomide, received 2 courses of bortezomib treatment. In the first course, bortezomib alone was administered and then in the second course bortezomib was given in combination with dexamethasone. The patient's serum IgG level decreased from 8040 to 1020 mg/dl and the level of plasma cells in bone marrow was 1.2% after the treatments. Adverse reactions including rash, anemia, and thrombocytopenia occurred in the first course; however, they were milder in the second course combined with dexamethasone. Case 2: A 43-year-old man with MM (IgD, gamma-type, stage IIA) resistant to conventional and high-dose chemotherapy with PBSCT as well as thalidomide therapy, received treatment with bortezomib alone and then in combination with dexamethasone. His serum IgD level decreased from 2140 to 623 mg/dl. He suffered adverse reactions such as fatigue, anemia, and thrombocytopenia in the first course, which were relieved in the second course. These results indicate that the combination of bortezomib and dexamethasone is effective in the treatment of refractory MM and that dexamethasone can reduce the adverse reactions of bortezomib.

Polycythemia vera (PV) is a hematopoietic stem cell clonal disorder, 5 to approximately 10% of which will evolve into acute leukemia. The pathophysiology of leukemic transformation and the best therapy for the leukemic phase of PV is still unknown. A 73-year-old woman was given a diagnosis of PV 17 years previously. However, laboratory data revealed myeloblasts in the peripheral blood with macrocytic anemia and thrombocytosis in March 2003. Bone marrow examination in October 2003 revealed 32.8% myeloblasts with trilineage dysplasia. Chromosomal analysis of the bone marrow cells revealed that 18/22 of mitotic cells had del(20q) and 3/22 of had t(2; 12). The leukemic phase of PV was diagnosed. She achieved not only a hematologic remission, but also cytogenetic remission after interferon (IFN)-alpha treatment. As far as we know, this is the first report of the introduction of IFN-alpha in the treatement of the leukemic phase of PV. Further monitoring of this patient will provide valuable information concerning the pathophysiology of leukemic transformation and the development of effective therapy for the leukemic phase of PV.

A 33-years-old man was diagnosed as having undifferentiated carcinoma presenting with right neck lymphadenopathy in December 2000. He obtained complete remission (CR) following chemotherapy, radiation and lymphadenectomy on the right neck. He had multiple para-aorta lymphadenopathy and splenomegaly in December 2001. An open-abdominal lymph node biopsy was performed from which a diagnosis of anaplastic large cell lymphoma was made. CR was achieved with biweekly CHOP, however, the patient suffered from a relapse twice. He underwent allogeneic peripheral blood stem cell transplantation (PBSCT) from his HLA-matched sister while in non-CR in November 2002. Engraftment was achieved on day 14, and at the same time, complete chimerism was confirmed. Acute grade III graft-versus-host disease (GVHD) developed and was controlled with cyclosporine A and prednisolone. Extensive chronic GVHD was subsequently observed and required systemic immunosuppression. His condition returned to CR after the PBSCT and he sustained complete chimerism. He suddenly died of fulminant thrombotic microangiopathy seven months after the PBSCT. The tumor cells were ALK-positive, CD30-negative and JH rearrangement was detected, and were therefore classified as diffuse large B-cell lymphoma with expression of ALK according to the WHO classification, though they differed from this subtype in some points. Although this case was refractory for chemotherapy with a complex karyotype, the graft-versus-lymphoma effect might have contributed to the sustained CR following the PBSCT.

A 59-year-old woman with goiter complained of nausea, vomiting and weight loss in April 2000. She underwent an endoscopic examination and was admitted to our hospital because gastric biopsy specimens revealed that she had diffuse large B-cell lymphoma. A thyroid biopsy also detected the diffuse infiltration of lymphoma cells, which were positive for CD19, CD20, CD38 and HLA-DR. Although the cells expressed surface immunoglobulin a chain, they lacked expressions of the kappa and lambda light chains. Chromosomal analysis of the thyroid cells showed 47, XX, t(2 ; 3)(q31 ; q13), + 3, t(8 ; 22)(q24 ; q11). After five courses of biweekly CHOP chemotherapy, she received autologous peripheral blood stem cell transplantation in October 2000. Currently, she has maintained complete remission for more than 4 years.

We present two cases of malignant lymphoma that developed a high fever that eventually reached an extremely high 38.9 degrees C. The C-reactive protein( CRP) elevation also climbed to a very high 12.2 mg/dl after treatment with granulocyte colony-stimulating factor (G-CSF). In the one case, after stem cells were mobilized with CHASER therapy (cyclophosphamide, cytarabine, etoposide, dexamethasone and rituximab) followed by G-CSF (filgrastim 600 microg/day) subcutaneous daily, the serum CRP level rose to a maximum of 5.6 mg/dl, with a maximum fever elevation of 38.9 degrees C. In the other case, after the subject was given CHASE therapy followed by subcutaneous treatment with G-CSF (filgrastim 75 microg/day) daily, the maximum serum CRP level was 12.2 mg/dl along with a maximum fever of 38.9 degrees C. Although no infection was found in either case, multiple antibacterial agents were ineffective;after discontinuation of G-CSF, fever dissipated and the serum CRP level became negative. G-CSF induces the proliferation and differentiation of neutrophils and also causes the mobilization of mature neutrophils from hematopoietic tissues. With the increasing propensity to G-CSF, we must keep in mind the possibility of such adverse reactions so as to serve the overall best interests of the patient.

A number of genes associated with life phenomena have been identified by the achievement of genome projects. As both comprehensive analysis and methods for investigation of specific genes have been developed, we can understand the pathogenesis of ocular diseases and develop novel medical treatments based upon detailed information on molecular mechanisms. In our review article, we focused on three vision-threatening ocular diseases; glaucoma, age-related macular degeneration (AMD) and proliferative vitreoretinopathy (PVR), and discussed the potential and problems related to retinal regenerative therapy. Regarding glaucoma, we investigated the relationships between aqueous humor and cell components in the aqueous outflow route. We have revealed that the Rho-Rho-associated coiled-coil-forming protein kinase (ROCK) signal transduction pathway participates in regulation of the aqueous outflow route, and that ROCK inhibitors and several protein kinase inhibitors exert intraocular pressure-lowering effects. Also, we conducted a series of investigations on familial amyloidotic polyneuropathy (FAP), as a representative secondary glaucoma caused by genetic mutations in a single gene. We reviewed the clinical features of ocular complications derived from FAP, their molecular mechanisms and possibilities for the development of novel medical treatments. In addition, we discussed a novel therapeutic concept, "neuroprotection", and showed the potential of some drugs as candidates for the neuroprotective treatment of glaucoma. Against AMD, we have performed a series of experiments from the viewpoint of similarity with atherosclerotic lesions. We have shown the molecular mechanisms of AMD associated with up-regulated expression of scavenger receptors and the interaction between leukocytes and vascular endothelial cells. Furthermore, in the pathogenesis of PVR, we described the role of epithelial-mesenchymal transition in retinal pigment epithelial cells and demonstrated the usefulness of enzymatic vitrectomy. Although retinal regenerative therapy has attracted much attention from global investigators, we pointed out its limitation for clinical application, and developed researches on efficient culture method using physiologically active factors for proliferating retinal stem cells with multi-potentiality, differentiation of the transplanted progenitor cells, and axon guidance of neurons by extracellular matrices.