Autologous peripheral blood stem cell transplantation (auto-PBSCT) is considered the standard therapy for younger patients with multiple myeloma, however, survival curve doesn't reach plateau. On the other hand, allogeneic reduced-intensity stem cell transplantation (RIST) with graft-versus-myeloma (GVM) effects is expected to be the curable therapy. We had three cases received RIST (one case as a salvage therapy for relapse after tandem auto-PBSCT, and two cases as an intentional RIST after achieving partial response by auto-PBSCT). All of three cases relapsing or progressing after RIST showed disease type of plasmacytoma without plasma cells in the bone marrow. That may be because conditioning regimen doesn't include total body irradiation, or GVM is ineffective in plasmacytoma, but effective in bone marrow. Here, we report clinical course of these three cases with some consideration.

More than 100 HPV genotypes are presently distinguished by comparing the DNA sequence of the L1 ORF of each HPV. Two important aspects of the nature of this group of heterogeneous viruses are the way in which specific HPV genotypes are associated with distinct clinical and histological morphologies and the way specific HPV genotypes affect distinct anatomical sites. The former is best evidenced by the HPV type specific cytopathic or cytopathogenic effect (CPE), whereas the latter is suggested by the marked preference of each HPV genotype for specific tissues and sites. Recent studies have also suggested that specific HPV genotypes may target epithelial stem cells at specific anatomical sites. HPV type-specific CPE is the central schema when we analyze and understand the HPV-associated diseases. The concept was suggested by the characterization of distinct HPVs from different types of warts: HPV 2/27/57 from common warts, HPV 3/10/28 from flat warts, HPV 6/11 from condyloma acuminatum, and HPV 5/8 from lesions of epidermodysplasia verruciformis (EV). In this paper, I summarize recent advances in HPV study field, especially on HPV-associated cutaneous lesions. These include inclusion warts, HPV-associated epidermoid cysts, HPV type specific activation of melanogenesis, a double infection with HPV 1 and HPV 63 within a single cell, primary target cells and life cycle of the virus, and the identification of novel genes that are associated EV. The HPV-associated cutaneous lesions thus pose important problems to be resolved in virology and human pathology.

A 23-year-old man had been treated for repeated pneumonia from the age of 16. After a diagnosis of eosinophilic pneumonia at the age of 17, he had been given antibiotics and steroids. At the age of 23, he was referred to our hospital. Although he improved rapidly in response to antibiotic medication, his chest X-ray shadows did not improve. Video-assisted thoracoscopic lung biopsy was performed. Fibrosis and invasion of inflammatory cells were seen on the air duct walls and foreign bodies that were thought to be food residue were detected. A diagnosis of chronic aspiration pneumonia was made pathologically. A brain stem tumor was detected on his brain magnetic resonance imaging, which was thought to be the cause of his mis-swallowing.

The field of cell-based transplantation has expanded considerably and is poised to become an established cardiovascular therapy in the near future. In this review, we will focus on endothelial progenitor cells (EPCs), which are immature cells capable of differentiating into mature endothelial cells (ECs). The major source of EPCs is the bone marrow (BM). BM-derived EPCs are mobilized into peripheral blood and recruited to the foci of pathophysiological neovascularization and reendothelialization, thereby contributing to vascular regeneration. Severe EPC dysfunction is an indicator of poor prognosis and severe endothelial dysfunction. Indeed, number of circulating EPCs and their migratory activity are reduced in patients with diabetes, coronary artery disease (CAD) or subjects with multiple coronary risk factors. Effective neovascularization induced by EPC transplantation for hindlimb, myocardial and cerebral ischemia has been demonstrated in many preclinical studies, and early clinical trials of EPC transplantation in chronic and acute CAD as well as Buerger disease indicate safety and feasibility of the cell-based therapies. Further development in cell processing technology for efficient isolation, expansion, mobilization, recruitment and transplantation of EPCs into target tissues are underway and expected to be tested in clinical trials in the near future.

To apply human mesenchymal stem cells (hMSC) to regenerative medicines, it is necessary to multiply hMSC in vitro in a short period. In addition, it is desirable that the medium which is used for the hMSC multiplication is not supplemented with the serum, because the addition of the serum has risks of infection. In this study, we cultured hMSC with three kinds of medium used for multiplying hMSC (DMEM, MSCGM, STK2) and compared hMSC proliferation in each medium. As a result, it was confirmed that hMSC proliferation was significantly higher in STK2 medium which is a novel serum-free medium developed for hMSC multiplication. Moreover, we compared the hMSC proliferation in these media under the environment that assumed bone reproduction. When we cultured hMSC in each medium with hydroxyapatite (HAp), the proliferative inhibition by HAp depended on the additive amount, and the degree of the proliferative inhibition was different among the media but the lowest inhibitory effect was observed in STK2 medium.

Recent clinical neuroimaging studies have suggested the morphological brain changes occur and progress in the course of alcoholism and depression. The abnormality of neurogenesis has emerged as a potential epidemiological mechanism of these diseases. Previously, we have indicated the low dose of ethanol that could not influence on the survival of neurons and neural stem cell (NSC) suppress differentiation to neurons but glias through activation of neuron-restrictive silencing factor/repressor element-1 silencing transcription factor (NRSF/REST). We revealed the endoplasmic reticulum function and trophic factor signaling change implicated in this mechanism of ethanol action on NSC differentiation change. The analysis of potentials of psychotropic drugs on the ethanol-induced NSC function change may reveal the possible biological way of neural network impairment and its repair. Furthermore, the approach of using stem cells such as intravenous NSC transplantation can be a useful method to clarify the neural network reconstruction damaged by ethanol. The importance of interactive analysis of in vitro to in vivo should be documented for the pathophysiological understanding and new therapy development against alcohol-induced brain damage.

A free radical scavenger Edaravone is the first clinical drug for neuroprotection in the world which has been used from 2001 in most ischemic stroke patients in Japan, and is especially useful in thrombolytic therapy with tissue plasminogen activator (tPA). Of great importance for regenerative therapy and gene therapy are the neural stem cells which are intrinsically activated or exogenously transplanted. Addition of NTFs greatly enhanced an intrinsic migration or invasion of stem cells into the scaffold, which could provide a future regenerative potential against ischemic brain damage at chronic stage.

The year of 2007 was a turning point of the treatment of Parkinson's disease (PD) in Japan. Severe adverse effects of dopamine agonists including valvular heart disease induced by ergots and sudden onset of sleep attacks induced by non-ergots, were disclosed, and treatments with agonists were reassessed. Good news were marketing of ropinirole, a new non-ergot agonist, in December 2006 and entacapone, the first catechol-O-methyl transferase (COMT) inhibitor in Japan in April 2007. Having faced these new situations, Japanese Neurological Association has started revising "the Guideline 2002 for the treatment of Parkinson's disease". Clinical trials of translational gene therapy for Parkinson's disease with adeno-associated virus (AAV) vector are now going on in four approaches: restoring dopamine synthetic capacity, protecting against cell death with trophic factors, interfering with the aberrant protein aggregation, and converting the subthalamic nucleus into an inhibitory, rather than an excitatory, structure. In Japan, gene delivery of the dopamine synthesizing enzyme aromatic amino acid decarboxylase (AADC) to the striatum of PD patients is going on in Jichi Medical University. New findings of the causative genes, environmental factors and molecular mechanism of PD have provided with new tools for developing new treatments. The big success of induction of induced pluripotent stem (iPS) cells from fibroblast has given an impact on cell therapy research of PD.

microRNAs (miRNAs) are non-coding RNAs, and are negatively regulating the gene functions through translational repression or digesting mRNAs by targeting 3'-UTR in messenger RNAs (mRNAs). To clarify roles of miRNAs in normal and pathological erythropoiesis, we analyzed the expression of miR-451, miR-155, miR-221 in normal human erythroid colony forming cells (ECFCs) and circulating red blood cells using quantitative real-time polymerase chain reaction. Remarkable down-regulation of miR-155 (about 200-fold) and up-regulation of miR-451 (270-fold) were observed during 12 days of ECFC cultures. miR-221 was down-regulated moderately (20-fold). Since mature erythroid cells expressed miR-451 at high levels, we analyzed the content of miR-451 in fractionated peripheral blood cells obtained from normal subjects (n= 3), and found that the expression level of miR-451 in red blood cells was about 10(4)-fold more than in granulocytes and mononuclear cells. Analysis of packed red cells from normal subjects (n=3) and patients with hematological disorders (n=16), showed that there was no significant difference of the expression levels of miR-451 between normal and patients. In conclusion, expression profiles of miR-155 and miR-451 are stage-specific during normal erythropoiesis in which miR-155 is down-regulated and miR-451 is up-regulated. A high expression of miR-451 is specific to red blood cells among circulating blood cells. These observations suggest that an analysis of microRNAs is a new relevant approach to understand underlying mechanisms of hematological disorders.

Hematopoietic malignancies include leukemia, lymphoma and multiple myeloma. These diseases are primarily diagnosed on the basis of morphological features of affected cells, which appear in peripheral blood, bone marrow and lymphoid organs. By taking advantage of the repetitive accessibility of the neoplastic cells within the peripheral blood / and/or bone marrow aspirates, morphological tests are conducted not only for diagnosis but also for evaluation of clinical outcomes and prognosis, suggesting that the morphological features are considered as a clinical biomarker in hematopoietic malignancies. However, remarkable progress in molecular targeted therapy and allogeneic hematopoietic stem cell transplantation has improved the long-term prognosis of patients with hematopoietic malignancies, and some patients are curable. Under such modern strategies for therapy, monitoring of minimal residual disease(MRD), which is morphologically undetectable, is required to guide proper management of the disease by evaluation of an optimal response to therapy and early detection of disease relapse. At present, both immunophenotypes(surface markers)and chimeric fusion genes(e. g. BCR-ABL in chronic myeloid leukemia)characteristic of hematopoietic malignant cells are analyzed as clinically useful biomarkers to monitor MRD by two highly sensitive methods, multiparameter flow cytometry and real-time quantitative PCR, respectively. On the other hand, serum markers reflecting the size of the tumor mass are clinically available to monitor the disease progression in mass-forming hematopoietic malignancies: e. g., soluble IL-2 receptor for lymphoma and M-protein or free light chain for multiple myeloma.

Faithful genome duplication is achieved by accurate coordination between DNA replication and chromosome segregation. Abnormalities occurring in this process are checked by biochemical signal transduction pathways, called checkpoints, which ensure the orderly progression of events in the cell cycle. Checkpoints prevent transition into subsequent phases until all processes in the previous phase are completed. Defects in cell cycle checkpoints result in gene mutations, chromosome damage, and aneuploidy, all of which contribute to tumorigenesis. However, it has recently been uncovered that the impairment of checkpoint function is the major reason why DNA damaging anti-cancer agents can selectively kill cancer cells. Given that G1 and G2 checkpoint functions are generally impaired in cancer cells, cells with DNA damage are unable to maintain G2 arrest and eventually die as they enter mitosis. This process is known as mitotic catastrophe.

Sh2b3/Lnk consisting of an N-terminal proline-rich region, PH-, SH2-domains and a tyrosine phosphorylation site, forms an intracellular adaptor protein family conserved from drosophila to mammals, together with Sh2b1/SH2-B and Sh2b2/APS (adaptor protein with PH and SH2 domains). Lnk negatively regulates lymphopoiesis and early hematopoiesis. The lnk-deficiency results in enhanced production of B cells, and expansion as well as enhanced function of hematopoietic stem cells (HSCs), demonstrating negative regulatory functions of Sh2b3/Lnk in cytokine signaling. Our recent studies also revealed that Sh2b3/Lnk functions in responses controlled by cell adhesion and in crosstalk between integrin- and cytokine-mediated signaling. Importantly, recent genome-wide association studies of the autoimmune type 1 diabetes or celiac disease identified risk variants in the SH2B3/LNK region, indicating possible unrevealed functions mediated by this adaptor molecule. This review summarizes roles of Sh2b3/Lnk in the regulation of B-lymphopoiesis and HSCs expansion and function, and briefly introduces our approach for modulating HSCs function by targeting Sh2b3/Lnk-mediated pathways.

Adult neurogenesis occurs in the olfactory bulb and the dentate gyrus of the hippocampus. It has been shown that exposure to psychosocial stress reduces cell proliferation in the dentate gyrus. However, little is known about how stress affects the proliferation kinetics of neural stem cells (NSCs) in the adult brain. We utilized a forced swim model of stress in the mouse and found that chronic stress decreased the number of NSCs. The reduction in NSC number persisted for weeks after the cessation of stress, but was reversed by treatment with antidepressant drugs, fluoxetine and imipramine. However, these antidepressants exhibit no direct effects on NSCs, suggesting that the effects of antidepressants on NSCs are mediated by serotonin. In contrast, mood stabilizing drugs, which are used to treat patients with bipolar disorder, act cell-autonomously on NSCs and enhance their self-renewal capability. Importantly, this enhancement is achieved at therapeutically relevant concentrations in the cerebrospinal fluid. The pharmacological effects are mediated by the activation of Notch signaling in the NSC, but not by the inhibition of GSK-3b signaling or inositol depletion, currently popular models to explain mood stabilizers' action. These data provide insights into the molecular mechanisms underlying the pathogenesis of mood affective disorders.

A 54-year-old woman visited our hospital with the chief complaints of generalized pain and thirst. Through hematologic and bone marrow examination, we diagnosed her illness as primary plasma cell leukemia. Three courses of VAD (vincristine, adriamycin and dexamethasone)therapy induced complete remission. Afterward, the patient received high-dose cyclophosphamide (4 g/m(2))for hematopoietic stem cell mobilization, and peripheral blood hematopoietic stem cells were collected 14 days later from the start of chemotherapy. Thereafter, the patient received 2 courses of high-dose melphalan (200 mg/m(2)), followed by tandem autologous peripheral blood stem cell transplantation. The patient developed no adverse events other than fever and stomatitis during the 2 transplantation courses. Fifty-six months after the transplantations, she remains in complete remission with continuous oral administration of bisphosphonate alone. Taken together, although primary plasma cell leukemia is a rare hematologic malignancy with poor prognosis, the intensive chemotherapy followed by tandem autologous peripheral blood stem cell transplantation may well improve the clinical outcome.

Ethanol is a deleterious agent that causes various kinds of neuronal damage to both the developing and adult brain. Recent research on alcoholism implicates impaired function of neural stem cell (NSC) in the pathogenesis of ethanol-induced brain dysfunction. We previously reported that the differentiation of NSCs into neurons was significantly influenced by ethanol. We also found that neuron-restrictive silencer factor/repressor element 1-silencing transcription factor (NRSF/ REST) binding activity potentiated by ethanol underlies the mechanism of ethanol inhibition of neuronal differentiation. Epigenetics refers to post-translational modifications of DNA and nuclear proteins that produce lasting alterations in patterns of gene expression. Epigenetic mechanism plays a critical role of neuronal plasticity and there is clear evidence that dysfunction of epigenetic mechanism also contributes to neurological and psychiatric illness. We will review epigenetic regulation in pathogenesis of psychiatric illness including alcoholism. We also demonstrated that trichostatin A, histone deacetylase inhibitor, reduced the ethanol-induced suppression of neuronal differentiation of NSCs. We suggest that ethanol alters the function of neural differentiation through the mechanism of potentiation of NRSF/REST binding and histone modifications.

A 26-year-old woman with acute lymphoblastic leukemia (ALL) relapsed three times after HLA-matched related bone marrow transplantation. Initially, ALL relapsed in the central nervous system (CNS) 1 year after transplantation. Then, ALL relapsed as a single bone tumor involving the CNS and pelvis 4 years after transplantation. Finally, multiple bone tumors in the pelvis and lumbar bones were found as well as spread to the bone marrow 5 years after transplantation. Bone marrow aspiration also showed ALL relapse. Flow cytometry analyses detected CD20-positive cells in the bone tumor. Though the initial bone tumor was resistant to hyper CVAD, radiation was effective and this patient achieved complete remission. At that time, the total radiation dose had already reached the upper limit. After the third relapse, bone marrow achieved complete remission with the administration of pirarubicin, vincristine, prednisolone, and L-asparaginase (arranged DVP-L), though this combination chemotherapy itself was not effective in multiple bone tumors. Thereafter, arranged DVP-L plus rituximab was administered, which resulted in significant tumor reduction. Biweekly rituximab administration as maintenance therapy has completely prevented the regrowth of bone tumors. Rituximab for relapsed CD20-positive ALL patients after stem cell transplantation could be beneficial.

Hemophagocytic lymphohistiocytosis (HLH) is a rare but severe complication of Epstein-Barr virus (EBV) infection. Interactions between EBV-infected T cells and activated macrophages cause several conditions such as pancytopenia, liver dysfunction and coagulopathy. We describe here two young adults with EBV-associated HLH with monoclonal proliferation of EBV-infected T cells within a short period after infectious mononucleosis as a primary infection. One patient was a 16-year-old man who developed severe pancytopenia and liver dysfunction two months after infectious mononucleosis. Bone marrow examination showed hemophagocytosis, and laboratory data demonstrated monoclonal proliferation of EBV-infected T cells. Several treatments such as immunosuppressive therapy, chemotherapy and hematopoietic stem cell transplantation were not effective, and the patient died of progressive disease. The other patient was a 19-year-old woman who developed thrombocytopenia and liver dysfunction two months after infectious mononucleosis. Findings of hemophagocytosis and monoclonal proliferation of EBV-infected T cells were similar to those in the first case. Clinical signs and symptoms were resolved completely by immunosuppressive therapy containing methyl-prednisolone and cyclosporine. Since these two cases each demonstrated a distinct clinical course, an investigation of the prognostic factors and treatment strategies for EBV-HLH is warranted.

We investigated mRNA expression of c-myc and chromosome aberrations at the c-myc locus in the same passage number of human mesenchymal stem cells (hMSCs). To understand the sensitivity of mRNA expression and the induction of chromosome aberrations, we first tested them in hMSC and cancer cell lines (HeLa S3, HOS, and OUMS-27). The c-myc mRNA expressions in HeLa S3 and OUMS-27 were significantly higher than those in hMSC, but then those in HOS were not. On the other hand, c-myc aberrant cells detected by fluorescence in situ hybridization in HeLa S3, HOS, and OUMS-27 were significantly higher than that in hMSC. Both analyses were performed in hMSCs derived from five donors for the culture period of 50 days. In hMSCs from one donor, the frequency of c-myc aberrant cells significantly increased at 20 and 50 days respectively, and each mRNA expressions had a tendency to increase, but there is no significant change among 3, 20 and 50 days. In hMSCs from the others, both endpoints did not change for 50 days. For safe use of somatic stem cells in the regenerative medicine, the investigation of characteristic change of them during the in vitro culture is important. In the present study, we showed the mRNA expressions and chromosome aberrations of hMSCs in in vitro culture as the first step for establishing of safety evaluation of tissue engineered medical devices using normal hMSCs.