Impact of posttransplant cyclophosphamide on the outcome of patients undergoing unrelated single-unit umbilical cord blood transplantation for pediatric acute leukemia.

Background: Umbilical cord blood transplantation (UCBT) from unrelated donors is one of the successful treatments for acute leukemia in childhood. The most frequent side effect of UCBT is peri-engraftment syndrome (PES), which is directly associated with the greater prevalence of acute and chronic graft-versus-host-disease (aGvHD and cGvHD). In haploidentical stem cell transplantation, posttransplant cyclophosphamide (PTCY) has been demonstrated to be an effective method against GvHD.

Modified conditioning regimen improves outcomes of unrelated donor peripheral blood stem cell transplantation for β-thalassaemia major patients.

Background: The objective of this study was to evaluate the feasibility of a modified conditioning regimen for the treatment of patients with β-thalassaemia major (TM), using unrelated donor peripheral blood stem cell transplantation (UD-PBSCT).

Methods: A modified conditioning regimen based on intravenous busulfan, cyclophosphamide, fludarabine, and antithymocyte globulin was performed in 50 consecutive childhood patients with β-TM and a median age of 4.6 years (range, 2-12 years).

LW-213 induces G2/M cell cycle arrest through AKT/GSK3β/β-catenin signaling pathway in human breast cancer cells.

LW-213 is a derivative of Wogonin and the anticancer activities of Wogonin have been reported. To study whether LW-213 inhibits cancer cells and explore a possible mechanism, we investigate the compound in several cancer cell lines. We found LW-213 arrests G2/M cycle in breast cancer cells by suppression of Akt/Gsk3β/β-catenin signaling pathway.

[Therapeutic efficacy evaluation of rabbit anti-thymocyte globulin combined with cyclosporine A in children with aplastic anemia].

This study was aimed to investigate the therapeutic efficacy of rabbit anti-thymocyte globulin (r-ATG) combined with cyclosporine A (CsA) and to analyse the efficacy-related factors in children with aplastic anemia (AA). Twenty five AA children treated with r-ATG [3.5 mg/(kg·d)×5 days] combined with CsA were analyzed retrospectively.

[Analysis of one case of adolescent blastic plasmacytoid dendritic cell neoplasm].

This study was purposed to summarize the clinical characteristics and laboratorial data of blastic plasmacytoid dendritic cell neoplasm (BPDCN) in pediatric patients in order to enhance understanding this disease in diagnosis and therapy. A rare case of BPDCN in children was enrolled in this study. The blood routine test, examination of bone marrow cell morphology, histopathology and immunophenotype of the skin lesions were performed and analysed, the single cell suspensions of the biopsied skin mass were detected by flow cytometry.

Rabbit-antithymocyte globulin combined with cyclosporin A as a first-line therapy: improved, effective, and safe for children with acquired severe aplastic anemia.

Purpose: Acquired aplastic anemia is an organ-specific auto-immune disease characterized by pancytopenia and hypoplastic bone marrow. Immunosuppression with anti-thymocyte globulin (ATG) and cyclosporine A (CsA) is an effective and safe therapy for patients without undergoing hematopoietic stem cell transplantation. The aim of the current study was to investigate the effect of rabbit-ATG (r-ATG) combined with CsA as an intensive immunosuppressive therapy (IST) for acquired severe aplastic anemia (SAA) in children.

[Sequentially inducting murine embryonic stem cells into hematopoietic stem cells in vitro by hematopoietic development procedure for reconstitution of hematopoiesis in vivo].

This study was purposed to directly induce murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) by simulating the spatial and temporal hematopoietic microenvironment changes in embryonic development, and to investigate the function of in vivo hematopoietic reconstitution of these HSC. E14 ESC were induced into embryoid body (EB) firstly. Then the cells from EB were further co-cultured with human aorta-gonad-mesonephros (AGM) region, fetal liver (FL) and bone marrow (BM) stromal cells in Transwell non-contact system in sequential orders.

[Effects of human aorta-gonad-mesonephros region stromal cells on inducing differentiation of murine embryonic stem cells into hematopoietic stem/progenitor cells in vitro].

This study was aimed to investigate the effect of human aorta-gonad-mesonephros (AGM) region stromal cells on differentiation of murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) in vitro and to clarify their effect mechanism. E14 murine ESC were induced into embryo body (EB) firstly. Then the EB cells were further co-cultured with the stromal cells from human AGM region, fetal liver (FL) or bone marrow (BM) in Transwell non-contact system.

Allogeneic hematopoietic stem cell transplantation for childhood aplastic anemia: prospective trial in China.

We aim to investigate the efficacy and safety of the treatment with fully matched allogeneic hematopoietic stem cell transplants for children with severe aplastic anemia (SAA) in the first prospective trial in China. Six SAA children received allogeneic hematopoietic stem cell transplantation combined with chemotherapy. Five patients had successful engraftment while the sixth child regained normal peripheral blood counts consistent with spontaneous autologous hematopoiesis.

Bone marrow mesenchymal stem cells reduce the antitumor activity of cytokine-induced killer/natural killer cells in K562 NOD/SCID mice.

Adoptive cellular immunotherapy is an important treatment to eliminate residual tumor cells after hematopoietic stem-cell transplantation. Bone marrow mesenchymal stem cells (MSC) have previously been shown to exert immunoregulation functions, including inhibition of proliferation and killing activities of T cells and natural killer (NK) cells in vitro and reduction of the graft-versus-host disease. MSC can survive in vivo for a long period of time, the influence of MSC on the antitumor activity of subsequently infused immune killer cells is not clear.

[Influence of FcγRIIIa polymorphism on rituximab-dependent NK cell-mediated cytotoxicity to Raji cells].

Fcgamma receptor IIIa (FcγRIIIa) polymorphisms is considered to influence clinical response to therapeutic monoclonal antibody (McAb) in cancer, most people believe it can affect McAb binding, and McAb-dependent NK cell-mediated cytotoxicity. This study was purposed to determine the difference of antibody-dependent cell-mediated cytotoxicity (ADCC) effects mediated by different FcγRIIIa NK cells. The FcγRIIIa genotypes were detected by nest-PCR, the target cells (Raji cells) were stained with 5- (and 6-) carboxyfluorescein diacetate succinimidyl ester (CFSE), cultured with effector cells with different FcγRIIIa genotypes, and finally stained with propidium iodide (PI); the CD20 expression of Raji cells were tested by flow cytometry and cytotoxic index was calculated as well.

Optimized protocols for generation of cord blood-derived cytokine-induced killer/natural killer cells.

The efficacy of various combinations of stem cell factor (SCF), FLT3 ligand, interleukin (IL)-2, IL-7 and IL-15 to induce and expand cord blood-derived cytokine-induced killer (CIK) cells was investigated. There were three treatment groups: group A: SCF combined with IL-2, IL-7 and IL-15; group B: SCF, FLT3 ligand combined with IL-2, IL-7 and IL-15, and group C: IL-2, IL-7 and IL-15, the control group. Proliferation rates of CD3(+)CD56(+) CIK cells and CD3(-)CD56(+) natural killer (NK) cells were highest in group B; expansion of CIK cells increased 796.

[Fcγ receptorIIIa polymorphism in healthy children and those with hematological malignancies].

Fcγ receptorIIIa (FcγRIIIa) polymorphism was considered to influence clinical response to therapeutic monoclonal antibody (MAb) against cancer, which is suggested to affect MAb binding and MAb-dependent NK cell-mediated cytotoxicity. The purpose of this study was to examine the FcγRIIIa gene polymorphisms in healthy children and in children with hematological malignancy, and to explore its possible effect on MAb in children with hematological malignancies. 43 healthy children (H) and 20 pediatric patients with hematological malignancies (HM) were enrolled in this study.

[Distribution and interaction of bone marrow mesenchymal stem cells and cord blood CIK/NK cells infused via different ways at different time periods in NOD/SCID mice].

The study was aimed to explore the distribution and interaction mechanism of human bone marrow mesenchymal stem cells (MSC) and cord blood cytokine-induced killer (CIK)/natural killer (NK) cells infused via different ways at different times in NOD/SCID mice. 5 microl 1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine perchlorate (DiI) dye(red) was added in suspension of MSC per ml, and 1 microl carboxyfluorescein diacetate, succinimidyl ester(CFDA SE) dye(green) was added in suspension of CIK/NK cells per ml. The amounts of MSC and CIK/NK cells infused in each 6 NOD/SCID mouse were 1 x 10(6) (0.

[Mesenchymal stem cells for treatment of steroid-resistant chronic graft-versus-host disease].

Objective: To assess whether treatment with mesenchymal stem cells (MSCs) is an effective adjunct therapy for refractory extensive chronic graft-versus-host disease (GVHD) resistant to conventional therapy.

Methods: 12 patients with steroid-resistant extensive chronic GVHD were treated with MSCs. One patient received one dose, 10 received two doses, and the remaining three doses.

[Effect of mesenchymal stem cells on expression of CD69 in cord blood CIK/NK cells and quantity ratio of T regulatory cells in CIK/NK cell culture].

This study was purposed to explore the effect of bone marrow derived mesenchymal stem cells (MSCs) on the expression of CD69 on cytokine-induced killer (CIK)/natural killer (NK) cells derived from cord blood and on the quantity ratio of CD4+CD25+ T regulatory cells in CIK/NK cell culture system using Transwell non-contact cell culture system. The experiments were divided into two groups: Transwell non-contact culture and mixture culture. The ratio of MSC to CIK/ NK cells was 1:20, 1:50 and 1:100.

[Relationship between cellular immune function during conditioning and graft rejection in patients with beta-thalassemia major].

Objective: To analyze the relationship between cell-mediated immune function during conditioning and graft rejection in patients with beta-thalassemia major.

Methods: Allogeneic hematopoietic stem cell transplantation was performed in 25 children with beta-thalassemia major and 11 with acute leukemia group. The percentages of T lymphocytes and natural killer (NK) cells in peripheral blood of these patients were detected with dual color immunofluorescence on day -10 (before conditioning) and day -5 (after conditioning), and the relationship between the cellular immune function and graft rejection was analyzed.

[Unrelated allogeneic umbilical cord blood transplantation: present status, problems and countermeasures].

As unrelated allogeneic umbilical cord blood transplantation (UCBT) has been developed for 20 years already since 1988, more than ten thousands cases have cumulatively undergone UCBT over the world. A huge number of clinical data confirmed that UCBT had unique characters with low rate of severe GVHD. The efficacy and data on TRM, relapse and EFS of allogeneic UCBT with HLA 0-1 mismatched are similar to those in HLA matched BMT.

[Effect of intra-bone marrow infusion of allogeneic mesenchymal stem cells on reconstruction of marrow mesenchymal stem cells in rat HSCT models].

This study was aimed to investigate the effect of intra-bone marrow (IBM) injection of allogeneic mesenchymal stem cells (MSCs) on reconstruction of bone marrow MSCs (BM-MSCs) in rats that received hematopoietic stem cell transplantation (HSCT), and to detect the donor MSCs in the hosts for clarifying the effect mechanism of donor MSCs. Wistar female rats conditioned with lethal dose 60Co gamma-ray irradiation were co-transplanted with F344 female fetal and neonatal peripheral blood (FNPB) and BrdU-labeled MSCs separated from bone marrow mononuclear cells of F344 male rats. The donor MSCs were infused by IBM injection in bilateral tibia or intravenous injection (IV), while the FNPB were all via IBM route.

[Analysis of lymphocyte immune abnormality in 52 cases of children idiopathic aplastic anemia].

This study was aimed to investigate the characteristics of lymphocyte immune abnormality in children idiopathic aplastic anemia (IAA) in order to explore the immune pathogenesis of childhood IAA. The phenotypes of lymphocytes, the ratios of Th1/Th2 and Tc1/Tc2, the levels of CD25(+) and CD4(+)CD25(+) T lymphocytes in peripheral blood of IAA patients were measured at the onset of disease by flow cytometry and were compared with that in normal controls. The influences of those immunological indicators on prognosis of IAA were also analyzed.

[BMP-4 and VEGF promote development of hematopoietic stem cells during the embryoid body formation from embryonic stem cells].

The objective of this study was to explore the effects of BMP-4 and VEGF on the development of primary hematopoietic stem cells during the differentiation of embryonic stem cells (ESCs) into embryoid body (EB). Murine E14 ESCs were seeded into semisolid methylcellulose-based medium for EB formation. According to added or not cytokines, experiments were divided into: (1) group of spontaneous differentiation without cytokine as control; (2) group of BMP-4 in different concentrations (0, 5, 15, 25 and 50 ng/ml); (3) group of BMP-4 combined with VEGF; (4) group of VEGF alone.

[Isolation and culture of human embryonic AGM derived HSPCs in hematopoietic culture systems created by AGM stromal cells].

This study was purposed to isolate human embryonic AGM derived HSPCs and investigate the effect of AGM stromal cells on AGM-derived HSPCs. Immunohistochemical sections of human AGM tissue were investigated for CD34, Flk-1 and VEGF expression. Human AGM-derived single cells were isolated and seeded onto pre-treated feeder of human AGM stromal cells (hAGMS3 and hAGMS4) by direct contact and non-contact co-culture in Transwell culture system.

[The cytotoxicity of CIK/NK cells stimulated by K562-DC fusion vaccines in NOD/SCID mice model for human erythroleukemia].

Objectives: To study the in vivo efficacy and the safety of cord blood derived CIK/NK cells stimulated by K562-dendritic cells (DC) fusion vaccines in NOD/SCID mice model for human erythroleukemia.

Methods: DC and CIK /NK cells were both derived from cord blood mononuclear cells. DC were fused with inactivated K562 leukemia cell by PEG to produce K562-DC fusion vaccines.