Dideoxynucleosides ; False Positive Reactions ; Fluorine Radioisotopes ; Fluorodeoxyglucose F18 ; Humans ; Inflammation ; diagnosis ; Neoplasm Staging ; Neoplasms ; diagnosis ; metabolism ; pathology ; radiotherapy ; Positron-Emission Tomography ; methods ; Radiotherapy, Intensity-Modulated ; Sensitivity and Specificity ; Tomography, X-Ray Computed ; Treatment Outcome

Hematopoietic Stem Cell Transplantation ; Humans ; Transplantation, Homologous ; Treatment Outcome

Acute Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; prevention & control ; therapy ; Postoperative Period ; Secondary Prevention

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

To study the biological characteristics of mesenchymal stem cells (MSC) from patients with myelodysplastic syndrome (MDS) and their supportive capacity for hematopoiesis in vitro, MSCs from bone marrow samples of MDS patients were isolated, cultured and expanded. Morphology, immunophenotype, osteoblasts differentiative and proliferative property of MSC and colony forming unit-fibroblast (CFU-F) were measured and analyzed. Mononuclear cells (MNC) of cord blood were plated onto a feeder layer formed by MSC of MDS patient, cells count and CFU-GM production were observed. The results showed that the culture-expanded cells from MDS patients presented a typical fibroblast-like morphology. Cells were positive for SH2 (CD105), SH3 (CD73), Thy-1 (CD90), but negative for CD34 and CD45. After induction, these cells could differentiate into osteoblasts. Their proliferative capacity and CFU-F number were similar to those of MSC from healthy donors. The total cell count and CFU-GM yield in supernatants after culture for 2 weeks were significantly lower than those of control in hematopoiesis supportive experiments in vitro (P < 0.05). It is concluded that the biological characteristics of MSC from bone marrow of MDS patients are not different from those of MSC isolated from bone marrow of normal donors, however, their capacity of hematopoiesis support in vitro are significantly weaker.
5'-Nucleotidase ; analysis ; Adult ; Aged ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; immunology ; Cell Differentiation ; Endoglin ; Female ; Hematopoiesis ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; immunology ; Middle Aged ; Myelodysplastic Syndromes ; blood ; Receptors, Cell Surface ; analysis

The interaction of killer cell immunoglobin-like receptors (KIR) with HLA molecules has particular relevance to the genetics, immune responses and allogeneic stem cell transplantation. The genes of KIR and HLA are located in different chromosomes and segregate independently. The repertoire of KIR molecules varies among NK cells and is determined by the KIR genotype. The HLA genotype has only subtle impact on the KIR phenotype. Three major HLA specificity groups are recognized by KIRs. Donor versus recipient NK-cell alloreactivity, when recipients lack HLA ligand for their donor inhibitory KIR, can benefit allogeneic stem cell transplantation, especially the HLA haploidentical hematopoietic stem cell transplantation. The outcome of stem cell transplantation can be best predicted by the presence of KIRs on the donor's NK cells and the absence of corresponding KIR ligand in the recipient's HLA repertoire-a receptor-ligand model. In this paper the interaction of KIR and HLA in hematopoietic stem transplantation, the genetic basis of KIR and HLA, the relation of KIR expression on NK cells with HLA and the role of KIR and HLA in immune responses were reviewed.
Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; genetics ; immunology ; Humans ; Killer Cells, Natural ; cytology ; immunology ; Receptors, Immunologic ; genetics ; immunology ; Receptors, KIR ; Transplantation Immunology

Spectral karyotyping (SKY) is a novel cytogenetic technique, has been developed to unambiguously display and identify all 24 human chromosomes at one time without a priori knowledge of any abnormalities involved. SKY discerns the aberrations that can not be detected very well by conventional banding technique and fluorescent in situ hybridization (FISH). So SKY is hyper-accurate, hypersensitive, and hyper-intuitional. In this paper the basic principle of SKY technique and its application in leukemia cytogenetics were reviewed.
Humans ; Karyotyping ; Leukemia ; genetics ; pathology ; Spectral Karyotyping

Non-myeloablative allogeneic hematopoietic stem cell transplantation (NMAT) is brought forward recent years. It is a modified transplant technology. NMAT treats malignant disease by graft versus tumor effect of chimerism after transplantation. The difference between this transplantation and standard allogeneic stem cell transplantation is the conditioning regimen. Non-myeloablative allogeneic stem cell transplantation increased the indication of allogeneic stem cell transplantation. This article focused on the background of this transplantation, the results of animal experimental and clinical research, the advantages and shortages of this transplantation.

Chronic graft-versus-host disease (cGVHD) continues to be a major complication in long-term survivors after allogeneic hematopoietic stem cell transplantation and is the principal cause of morbidity and non-relapse mortality. With the new approaches in the immune mechanisms of cGVHD, the diagnosis, prophylaxis and treatment of cGVHD are involved. This review summarises the current progress of research on cGVHD.
Chronic Disease ; Female ; Graft vs Host Disease ; diagnosis ; etiology ; therapy ; Humans ; Male ; Prognosis ; Risk Factors

Recombinant human granulocyte-colony-stimulating factor (rhG-CSF) can widely regulate human immunologic response. In the protocol of peripheral blood stem/progenitor cell mobilization, rhG-CSF can change the numbers and functions of T cells. Then the results can impact the incidence of graft-versus-host disease after allogeneic peripheral blood stem/progenitor cell transplantation. The regulation of rhG-CSF on T cell is an indirect action which is based on the direct action to monocytes and dendritic cells. The numerous IL-10 secreted by monocytes plays a key role in cytokines production, proliferative response and cytotoxicity of T cells. Endogenous IL-10 can induce high expression of SOCS3 and the SOCS3 is very important for regulating the signal transduction of the activities of T cells. In this review influences of rhG-CSF on T-cells in mobilization process and related mechanisms were elaborated with emphasis.
Blood Donors ; Granulocyte Colony-Stimulating Factor ; pharmacology ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Interleukin-10 ; biosynthesis ; Recombinant Proteins ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; biosynthesis ; T-Lymphocytes ; cytology ; drug effects ; metabolism