The myelodysplastic syndrome (MDS),which are characterized by the presence of ineffective hematopoetic stem cells.Research on pathogenesis and related epigetics has made significant progress in recent years,more and more abnormal cytogenetics and gene mutations are used for prognosis and treatment options for MDS.In the 56th American Society of Hematology (ASH) annual meeting,lots of gene mutations were introduced.

Relapse/refractory multiple myeloma must be the main tough for the current clinical therapy,thus at the 56th American Society of Hematology annual meeting the latest treatment studies for the disease in 2014 were described in detail,including the joint application of drugs,the new drug clinical research,the safety and efficacy that could be inspiring.This chapter shows the mechanism of these new drug action,as well as the methods and side effects.

Objective To detect and separate the side population cells(SP) from multiple myeloma (MM) cell lines PRIM8226,and to study their biological characteristics.Methods Fluorescence activated cell sorter (FACS) and Hoechst33342/PI dye were used to sort SP cells of PRIM8226.The multiplication capacity was tested by the growth curve and MTT test,SP cells proportion and the cell cycle were analyzed by flow cytometry,the colony-formtion ability was compared in terms of colony forming experiment,the expression of c-myc,KIF4,SOX2,OCT4 was tested by RT-PCR.The oncogenicity of the cells was analyzed by nude mouse transplantation tumor experiment in vivo.Results The ratio of SP cells in PRIM8226 was (1.78±0.89) ％.More SP cells in the G0 / G1 period,(44.34±3.09) ％ vs (28.49±1.97) ％,P ＜ 0.05,and fewer cells in S phase than MP cells,(38.83±3.69) ％ vs (51.49±4.62) ％,P ＜ 0.05.There were no difference in the expression of CD38 and CD138 between SP cells and MP cells,respectively,(78.5±8.5) ％ vs (82.0±4.0) ％ and (72.3±15.7) ％ vs (84.3±11.9) ％,P ＞ 0.05.Colony formation assay showed that the colony forming efficiency of the SP cells was higher than the MP cells,single cell clone diameter,the number of clone forming,the clone formtion rate were significantly higher than MP cells,0.280±0.016 vs 0.118±0.019,1 722±127 vs 358±14,(86.1±3.46) ％ vs (17.9±1.88) ％,P ＜ 0.05.The mRNA expression levels of c-myc,KIF4,SOX2,OCT4 in SP cells were higher than those in MP cells,c-myc (29.90±3.73) ％ vs (16.84±2.35) ％,KIF4 (29.97±2.89) ％ vs (19.06±1.23) ％,SOX2 (40.00±4.58) ％ vs (16.62±2.09) ％,OCT4 (32.96±0.92) ％ vs (23.27±0.92) ％,all P ＜ 0.05.Nude mouse transplantation tumor experiment in vivo showed the oncogenicity of the SP cells was more stronger than MP cells (5×103 vs 5×l05).Conclusion There are notably difference in the cell cycle,colony formation assay,the mRNA expression levels and oncogenicity,but no difference in the expression of CD38 and CD138,the proliferation ability between SP cells and MP cells.

Objective:To investigate the effects of dendritic cells(DCs)in pathogenesis of aplastic anemia,especially its roles in T cell abnormal activation.Methods:Direct immunofluorescence assays and flow cytometry(FCM)were used to determined proportion and kinds of DCs in bone marrow sample of patients with AA.The DCs were got from standard method by culturing mononuclear cells isolated from peripheral blood of normal person in medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and interleukin 4(IL-4)in vitro.The phenotype of DCs were determined by FCM.After pulsed with purified cord blood CD34+ cells and stimulated with recombinant human CD40 ligand(rhCD40L),the ability of those cultured CDs to activate and promote auto T lymphocytes to proliferation was tested.The changes of TCR V? gene repertorie of those T cells were also investigated by real time quantitative PCR(RQ-PCR).Results:The CD1a+CD11c+ and CD11c+CD83+ double positive cells increased in bone marrow of patients of AA,especially the proportion of CD11c+CD83+ cells increased significantly in those patients.After primed with CD34+ cells,DCs induced from peripheral monocytes could activate T lymphocytes and promote T cells to proliferation.The restricted usage of TCR V? genes was confirmed by RQ-PCR in those activated T cells by DC as those in AA patients.Conclusion:DCs,especially with phenotype of CD11c+CD83+,may have great impact on oligoclonal expansion of lymphocytes in patients AA,but the antigen which recognized by auto T cells and stimulated it to proliferation are still needed to further investigation.

Objective: To evaluate the incidence, clinical features, diagnosis and treatment of primary ex-tranodal non-Hodgkin' s lymphoma (PE-NHL). Methods: The data of 110 patients diagnosed as PE-NHL be-tween January 2001 and May 2008 were reviewed. Results: These PE-NHL patients counted 60.11% of the 183 malignant lymphoma patients at the same period. The primary sites affected were the gastrointestinal tract 21.82% (24/110), Waldeyer ring 10.91% (12/110), nasal cavity 9.10% (10/110), soft tissue 9.10% (10/110), mediastinum 7.27% (8/110) and other unusual sites 41.82% (46/110). Symptoms and signs of PE-NHL were not specific, and 77.27% of these cases had a swelling organ or lump of the primary organ affected. Ac-cording to the International Prognosis Index (IPI), the percentage of patients in low, intermediate, and high group was 41.11%, 44.44% and 14.44%, respectively. Immunophenotype was assayed in 93 cases. The per-centage of B-cell lymphoma was 69.90% while that of T-cell lymphoma was 30.10%. For those 95 cases treat-ed, the effective rate including complete remission (61.05%) and part remission (16.84%) was 77.89%, the median survival was 30 months, and the 5-year overall survival (OS) was 27%. While, for patients with prima-ry gastrointestinal non-Hodgkin' s lymphomas (PGIL), the complete remission rate, part remission rate and the effective rate was 65.21%, 17.39% and 82.80%, respectively. The median survival was 24 months, and the 5-year overall survival (OS) was 30%. Conclusion: PE-NHL is more common than nodal lymphoma. The symptoms and signs of PE-NHL of different sites are quite different. To improve the curative strategies of PE-NHL, it is important to make an allround understanding of PE-NHL and follow reasonable mode of diagno-sis and therapy.

BACKGROUND:Irradiation-induced skin injury is difficult to treat and easy to recurrence.Thus,irradiation-induced skin injury has been a difficulty in clinic.With the development of stem cell engineering and tissue engineering,practical application of bone marrow mesenchymal stem cells(BMSCs) has received much attention.In particular,the potential of multi-directional differentiation displays the application prospect in field of trauma repair.OBJECTIVE:To explore the effect of BMSCs on repair of irradiation-induced acute skin injury.DESIGN,TIME AND SETTING:The randomized,grouping design,comparison study was performed at the Laboratory of Hematopathy,Second Affiliated Hospital,Soochow University from June to August 2008.MATERIALS:A total of 46 male Kunming mice,weighing(25?2) g were used.Of them,40 were used to establish models of irradiation-induced local skin injury at grade Ⅲ.METHODS:A total of 40 mice were randomly selected and anesthetized with 3.6% chloral hydrate.Mouse skin of the buttock(2.0 cm?1.5 cm) was radiated using 4 MeV electron beam(1 Gy/min),total dose of 40 Gy.Mice were randomly assigned to experimental and control groups(n = 20).Mice in the experimental group were injected with CFDA SE-labeled BMSCs(0.1 mL)(2?1010/L) through the caudal vein immediately following irradiation.Mice in the control group were treated with an equal volume of saline.The remaining 6 rats were not radiated,but infused with an equal volume of BMSCs through the caudal vein.MAIN OUTCOME MEASURES:Wound surface injury in mice,pathological changes were observed 2 and 3 weeks following irradiation in the experimental and control groups.New vessels of skin were analyzed using immunohistochemistry.BMSC distribution in mouse skin and other tissues was observed at various time points in the experimental group.RESULTS:Following BMSC transplantation through the caudal vein,the speed of skin healing was faster 4-6 days in the experimental group compared with the control group.The trauma area was smaller in the experimental group compared with the control group,with the pathological presence of mild damage in epidermis,hair follicle,sesbaceous glands and collagen fiber.There were many new blood capillary in skin wound surface.BMSCs distributed widely in injured skin and multiple organs,and the number of BMSCs was significantly greater in the experimental group compared with the control group.CONCLUSION:BMSCs promoted the regeneration of irradiation-induced acute skin injury,which might contribute the interaction between BMSCs and wound microenvironment.

Objective To investigate the clinic effect of using three-dimensional conformal radiotherapy combining with Sanyangxuedai in treating advanced non-small cell lung cancer.Methods Grouping 100 patients with advanced non-small cell lung cancer randomly into a control group and a treatment group,with 50 patients in each.The control group was treated with three-dimensional conformal radiation therapy,the treatment group was treated on the basis of the control group with a ioint Sanyangxuedai.The clinical effects were compared in the two groups.Results The three-dimensional conformal radiotherapy combining with Sanyangxuedai had more better treating indications than using three-dimensional conformal radiotherapy only and the results had statistical significances(χ~2=12.01,P＜0.05).Conclusion The three-dimensional conformal radiotherapy combining with Sanyangxuedai had beaer clinical efrect in treating advanced non-small cell lung cancer.

Objective To deplore the immunoregulatory function changes of mesenchymal stem cells(MSCs)from multiple myeloma(MM)patients and its effects on the pathogenesis of myeloma bone disease.Methods MSCs from MM patients and normal controls were isolated and the immunophenotype was detected.Real-time PCR was performed to detect the expressions of TGF-β1,TGF-β2,TGF-β3,IL-6,IL3,TNF-α,FasL and RANKL of MSCs.Transwell coculture systems were performed between MSCs and T cells.Lymphocyte proliferative assay was employed to detect the effect of MSCs on T cell proliferation.The effect of MSCs on T cell cycle and T cell activation markers CD25 and CD69 expression were analyzed by flow cytometry.Cleaved caspase 3 protein by western blot and hoechst 33258 staining were employed to detect the apotosis of T cells.Influence of T cells on the osteogenesis potential of MSCs were detected by Von kossa stain,real-time PCR and Western blot.Results MSCs from both MM patients and normal controls possessed similar morphology and immunophenotypes.MM derived MSCs exhibited increased expressions of TGF-β1,IL-6,IL-3,TNF-α and RANKL and decreased expression of TGF-β2,TGF-β3 and FasL.The inhibitory effect of MM derived MSCs on T cell proliferative ability was attenuated compared to control MSCs.MSCs from normal controls silence more T cells in Go/G1 phase than those from MM patients.The daupening effect of MM derived MSCs on activation-induced T apoptosis seemed to be enhanced.Expression of T cell activation markers were significantly inhibited by MSCs from normal controls.Both T cells cocultured with MM deprived MSCs and T cells directly from MM patients inhibited osteogenesis potential of MSCs from normal controls.Conclusion MSCs from MM patients showed impaired immunoregulatory capability on T cells.The activated T cells,in turn,inhibited the osteogenesis potential of MSCs.This may participate in the pathogenesis of myeloma bone disease.

Objective To discuss function of the fusion cells of human bone marrow stromal cells (BMSCs) and human myeloma cell RPMI 8226 in the pathogenesis of multiple myeloma bone disease.Methods The cells,labeled by cell tracer green fluorescent probe (CMFDA) and red fluorescent probe (CMTMR),respectively,were induced into fusion by chemical polyethylene glycol (polyethyleneglycol,PEG-1000),and cell fusion model was set up.Whether fusion cells had nucleus fusion was determined by Karyotype analysis.The expressions of stemness-related genes,SIRP α gene and DC-STAMP gene in fusion cells were identified.Results Polyethylene glycol (PEG-1000) could mediate the integration of BMSCs and RPMI 8226 cells.The number of chromosomes in more than 80 ％ the hybrid cells was about 80.Fusion cells not only showed that BMSCs,stemness-related genes of c-myc,Klf-4 and OCT-4 genes expressed positively,but also the fusion-related genes SIRPα and DC-STAMP expressed positively.Conclusion BMSCs and RPMI 8226 cells can form fusion cells,and the cells have the potential for further integration,which is one of the important reasons for the promotion of muhiple myeloma bone destruction.

Objective To investigate the changes of follicular helper T cells (Tfh cells) and Tfh cells associated molecules in the peripheral blood (PB) of patients with malignant lymphoid diseases (MLD) dynamically, and explore their roles on pathogenesis of the diseases. Methods Fifty-five patients with MLD were enrolled in this study,including 9 patients with acute lymphocyte leukemia (ALL), 30 patients with non-Hodgkin lymophoma (NHL) and 16 patients with multiple myeloma (MM), and 10 healthy controls (NC) of similar age were also enrolled. The percentage of CD4+CXCR5+cells (Tfh cells) and expression of ICOS+, PD1+among the T cells were detected by flow cytometry (FCM), while the levels of interleukin 21 (IL-21) in plasma were detected by ELISA tests. Results The percentage of Tfh cells and expression of ICOS and/or PD-1 in PB of all untreated patients were significantly higher than those of NC (all P< 0.01), and MM group< ALL group 0.05), and apparently lower than those who achieved PR (P< 0.05). The percentages of Tfh cells in patients with T-ALL and T-NHL were higher than those in patients with B-ALL and B-NHL (P> 0.05), and much higher than NC (P< 0.01). The concentration of IL-21 in patients were much higher than that in NC [(326.56±32.44) pg/ml] (P<0.01), and MM group