BACKGROUND:Under mitomycin C treatment, feeder cells appear to have restricted proliferation, but they are stil able to secret different cytokines. Non-mesenchymal stem cells from the bone marrow and secreted factors in plasma maintain the micro-environment suitable for the growth of mesenchymal stem cells that can improve the yield of mesenchymal stem cells.
OBJECTIVE:To study the biological characteristics of C57 mouse bone marrow mesenchymal stem cells isolated using a whole bone marrow adherent culture technique.
METHODS:Using the whole bone marrow adherent culture technique, purified and amplified C57 mouse bone marrow mesenchymal stem cells were harvested. cellproliferation kinetics, immune cellsurface markers, multiple differentiation potential and cellcycle were detected.
RESULTS AND CONCLUSION:Using the whole bone marrow culture, mouse bone marrow mesenchymal stem cells were harvested and capable of adhering to the plastic culture vessel. The obtained cells expressed CD45, CD105 and Sca-1, but were negative for CD34, CD33 and C-kit. The doubling time was (57.11±1.5) hours. The cells could be induced to differentiate into osteoblasts, adipocytes and chondrocytes. The cellcycle analysis showed that 64%of cells were in G 0-G 1 phase. These indicates that C57 mouse bone marrow mesenchymal stem cells isolated using a whole bone marrow adherent culture technique have biological characteristics of mesenchymal stem cells.

BACKGROUND:Cryopreservation of human umbilical cord mesenchymal stem cel s has been a hot research issue currently, but the studies concerning their effects on expansion of hematopoietic stem/progenitor cel s after cryopreservation are seldom. OBJECTIVE:To investigate the effects of human umbilical cord mesenchymal stem cel s before and after cryopreservation as feeder layer on expansion of human bone marrow mononuclear cel s in vitro. METHODS:2.5g/L mitomycin C processed human umbilical cord mesenchymal stem cel s and bone marrow mesenchymal stem cel s at passage 3 were used as the feeder layer to expand adult al ogeneic bone marrow mononuclear cel s in culture. Up to day 35, methylcel ulose assay was used to detect hematopoietic stem/progenitor cel colony proliferation. RESULTS AND CONCLUSION:There were no differences in the morphology and size of colonies in the cryopreserved human umbilical cord mesenchymal stem cel group, bone marrow mesenchymal stem cel group and non-cryopreserved human umbilical cord mesenchymal stem cel group. However, these parameter described above were significantly higher in these three groups than the blank control group (P<0.05). There were fewer colonies in the cryopreserved human umbilical cord mesenchymal stem cel group than the non-cryopreserved human umbilical cord mesenchymal stem cel group (P<0.05). These findings indicate that human umbilical cord mesenchymal stem cel s before and after cryopreservation have the ability as feeder layer on expansion of bone marrow mononuclear cel s in vitro similar to bone marrow mesenchymal stem cel s. But this ability of human umbilical cord mesenchymal stem cel s may decrease after cryopreservation.

Background and purpose:T-cell non-Hodgkin’s lymphoma(T-NHL) has relatively high incidence in Asian countries, and the incidence rate showed an upward tendency. It has a dual characteristic of both ethnic and regional. We conducted this study in order to analyze the clinical characteristics, pathological type and survival of T-NHL in our hospital. Methods:Records of 50 cases with T-NHL treated from Jan. 2002 to Dec. 2012, were analyzed in terms of clinical characteristics, distribution of pathological type and survival. Results:Patients with T-NHL account for 24%of NHL cases in our study. Of the 50 cases, 37 were Han, 13 were Uygur;The most frequent type was NK/T cell lymphoma(NK/TCL)(16/50, 32%). We performed pathological type for each age group:4 cases with children’s group were all T-lymphoblastic leukemia/lymphoma(T-LBL)(4/4,100%), the most frequent type of youth and middle age group was NK/TCL(8/20, 40%;7/13, 53.8%), as the senile group was angioimmunoblastic T-cell lymphoma(AITL)(6/13, 46.2%). There was also a signiifcant difference in pathological type between the stage, with stageⅠ-Ⅱwere all NK/TCL(12/12, 100%), stageⅢ-Ⅳwere observed with peripheral T-cell lymphomas-unspeciifed(PTCL-U)(9/38, 23.7%) and AITL(9/38, 23.7%). The frequent type of Han was NK/TCL(14/37, 37.8%), as the Uygur was T-LBL(5/13, 38.5%). The median overall survival(OS) time was 12 (ranged 0-112) months. The 5 year OS rate was 39%. Both pathological type and age group were important factors influencing survival. The difference in outcome for the pathological type(P<0.05): NK/TCL and anaplastic large cell lymphoma(ALCL) had the better 3 year OS rates(71%, 61%), but PTCL-U and T-LBL had poor prognosis(19%, 7%). Age did a signiifcant effect on OS(P<0.05):children’s group had the worse 5-year OS rate (0%), middle age group had a better prognosis (67%), the OS of the youth group was 35%, for the senile group was 21%. Conclusion:T-NHL in Xinjiang region have their unique clinical characteristics:The overall incidence rate was similar with the domestic report, however, higher than the reports abroad. The incidence of Han was higher than Uygur. Pathological type showed in this study was different from that in European and American countries. There was a significant difference in distribution of pathological type in different age group, stage and nation. The long-term survival and prognosis of patients in Xinjiang region was poor. Both pathological type and age group were important factors inlfuencing survival.

Objective To observe clinical curative effect of the FLAG regimen combined donor lymphocyte infusion after granulocyte colony stimulating factor(G‐CSF) mobilization(G‐DLI) ,for the acute myeloid leukemia (AML) of allogeneic Peripheral blood hematopoietic stem cell trans‐plantation (allo‐HSCT) after recurrence of hematology .Methods For the patients with recur‐rence after allo‐HSCT ,giving the FLAG regimen chemotherapy when the WBC dropped to the lowest point ,followed by giving G‐DLI that infusion peripheral blood stem cell from the original donors ,to observe curative effect and survival situation .And searched the literature review through the PubMed etc .Results Through FLAG regimen combined G‐DLI ,3 cases of relapse after transplan‐tation again obtained complete remission (CR) .Case 1 :disease‐free survival (DFS) was 13 month and overall survival(OS) was 23 months after G‐DLI .The patient has been the central recurrence and remission in bone marrow ,he was dead after 23 months due to multipleorgan function failure .He occurred Ⅱ acute GVHD in Skin and Ⅰ acute GVHD in liver after G‐DLI and obtained effective control ,not chronic GVHD .Case 2 :DFS and OS were 12 months and 13 months ,as bone marrow relapse again and giving up treat‐ment ,so died a month later .Respectively ,he has limitations chronic GVHD in skin after G‐DLI .Case 3:DFS was 16 months after G‐DLI since the disease‐free survival ,had limitations GVHD in skin that was control for given small dose of immunosuppressive drugs .Conclusion Joint FLAG scheme and G‐DLI may be one of the effective treatment of postoperative recurrence of allo‐HSCT .

Objective: To analyze the efficacy of HLA-haploidentical peripheral hematopoietic stem cell transplantation (haplo-PBSCT) following reduced intensity conditioning (RIC) regimen to treat the patients with hematological malignancies who were older than 50 years old. Methods: Eighteen patients with hematological malignancies over 50 years were enrolled, including 8 male and 10 female patients. The median age of all patients was 52 (range: 50–66) years. Of them, 8 patients had acute myeloid leukemia (AML) , 2 chronic myelocytic leukemia (CML) , 5 myelodysplastic syndrome (MDS) , 2 acute lymphoblastic leukemia (ALL) , and 1 aggressive natural killer cell leukemia (ANKL) . All patients received fludarabine, cytarabine and melphalan with rabbit anti-human thymocyte globulin (FAB+rATG regimen) and transplanted with high dose non-T cell-depleted peripheral hematopoietic stem cells from donors. Enhanced graft versus host disease (GVHD) prophylaxis and infection prevention were administered. Results: Fifteen days after transplantation, 16 patients achieved complete donor chimerism. One of them rejected the donor graft completely at thirty days after transplantation, and the other 2 patients had mixed chimerism 15 days after transplantation and converted to complete recipient chimerism at 30 days after transplantation. The cumulative incidence of acute GVHD (aGVHD) was 61.1% (95%CI49.6%-72.6%) . The incidence of grade Ⅱ-Ⅳ aGVHD was 35.4% (95%CI 21.1%-49.7%) , whereas grade III-IV was 13.8% (95%CI 4.7%-22.9%) . The 2-year cumulative incidence of chronic GVHD (cGVHD) rate was estimated at 38.2% (95%CI 25.5%-50.9%) . Patients were followed-up for a median of 14.5 months (range, 3-44 months) . The Kaplan Meier estimates of 2-year overall survival (OS) and disease-free survival (DFS) was 72.6% (95%CI 60.1%-85.1%) and 63.7% (95%CI 49.2%-78.2%) , respectively. The 2-year cumulative incidence of relapse and non-relapse-mortality (NRM) was 31.2% (95%CI 16.5%-45.9%) and 12.5% (95%CI 4.2%-20.8%) , respectively. Conclusion RIC-haplo-PBSCT protocol can achieve better results in patients with hematologic malignancies over 50 years old.

OBJECTIVETo study the clinical features of acute graft-versus-host disease (aGVHD) and its risk factors for the related HLA-haploidentical non T cell-depleted in vitro peripheral hematopoietic stem cell transplantation (RHNT-PBSCT).

METHODSFrom July 2002 to December 2012, 104 patients who underwent the RHNT-PBSCT were enrolled to analyze the incidences, location and its risk factors of aGVHD, compared with those of the 103 patients who received the HLA-matched sibling non T cell-depleted in vitro PBSCT (MSNT-PBSCT) in the same period.

RESULTS(1)The cumulative incidence of aGVHD in the RHNT-PBSCT group was significantly higher than the MSNT-PBSCT group [(56.2±4.7)% vs (34±3.6)%, P<0.05], but the cumulative incidences of II-IV and III-IVgrade aGVHD had no significant difference between the two groups[(39.5±2.9)% vs (21.2±5.4)%, P>0.05; (12.6±4.1)% vs (10.8±2.4)%, P>0.05]. (2)The cumulative incidence of cutaneous aGVHD was significantly higher in RHNT-PBSCT group than that in MSNT-PBSCT group [(42.3±3.2)% vs (17.5±2.3)%, P<0.05]. The cumulative incidences of liver and gastrointestinal aGVHD between the two groups had no significant difference [(7.7±2.1)% vs (12.6±3.4)%, P>0.05; (16.3±4.5)% vs (10.3±2.5)%, P>0.05]. (3)The 3-year disease free survival (DFS) and overall survival(OS) of RHNT-PBSCT group and MSNT-PBSCT group were (63±5.5)%, (65.2±4.7)% and (74.2±5.4)%, (77.4±5)% respectively, without significance (P=0.078, P=0.052). (4)aGVHD occurrence with HLA haplotype (P=0.003) and matched loci (P=0.002) were significantly correlated by univariate analysis. Multivariate analysis showed that only the HLA typing is a risk factor for aGVHD (HR=1.891, P=0.03).

CONCLUSIONAlthough the incidence of total aGVHD in RHNT-PBSCT protocol is higher than that in MSNT-PBSCT, but there was no significance in severe aGVHD and cutaneous aGVHD was the common type, which indicates that RHNT-PBSCT protocol is feasible.

Disease-Free Survival ; Graft vs Host Disease ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; Humans ; In Vitro Techniques ; Incidence ; Peripheral Blood Stem Cell Transplantation ; Risk Factors ; Siblings ; T-Lymphocytes