Objective To investigate the effect of 1,25-(OH)2D3 and salvia miltiorrhiza on T cell subsets and cytokines in rats after allogeneic bone marrow transplantation.Methods Female Wistar rats were used as recipients and male SD rats were used as donors.All Wistar rats were divided into aGVHD group and intervention groups at random,and the intervention groups were further divided into 1,25-(OH)2D3 group,salvia miltiorrhiza group and combination group.The changes of T cell subsets (CD4+ and CD8+) and cytokine (IL-2,IFN-?,IL-4 and IL-10) in every group were detected and measured.Results When the presentation of aGVHD was relatively conspicuous,CD4+ and CD8+ were increased,and the increase of CD4+ was predominant.Compared with the difference prior to transplantation,the difference was statistically significant (P

Objective To observe the influences of 1,25-(OH)2D3 and salvia miltiorrhiza on hematopoiesis reconstruction and T cell subsets after allogeneic bone marrow transplantation in rats.Methods Donor rats were divided randomly into four groups,namely,Group A: aGVHD group;Group B: 1,25-(OH)2D3 group;Group C: salvia miltiorrhiza group;and Group D: two-drug combination group.We observed the survival time,peripheral blood cells,clinical manifestations of aGVHD and the changes of rat T cell subsets.Results Compared with those in aGVHD group,in every drug intervention group,the time for illness attack of aGVHD was delayed,aGVHD scoring was reduced and the average survival time was obviously prolonged(P

Objective To investigate the immunologic classification in the patients with acute leukemia (AL) in Xinjiang of China. Methods A panel of monoclonal antibodies (MOAb) and indirect immunofluorescence assay by fluoromicroscope was used to determine the pretherapy immunophenotype of 450 AL. Results 106 cases of acute lymphoblastic leukemia (ALL), 334 cases of acute myelogenous leukemia (AML), and 10 cases belonged to FAB unclassified acute leukemia (UAL) were unalysed. The expression of myeloid antigens in of ALL was seen in 15 % of 106 cases, and lymphoid-associated antigens were expressed in 25 % of 334 AML cases. The most frequently expressed antigen was CD7. The expression of myeloperoxidase (MPO) gene in 295 cases of AL were studied. The expression of MPO gene was observed in positive one of 81 ALL cases, and myeloid cells had different expression for MPO gene. Of the 9 cases of UAL, 6 cases were positive for MPO gene. There were no statistic differences of the expressions of the ALL stages between Han and Wei nationality. The order of myeloid markers expression in AML was as follows: CD_(33)>CD_(13)>CD_(15) inthe Han nationality, and the order of myeloid markers expression in AML was displayed CD(15)>CD(33)>CD_(14) in Wei nationality. Conclusion Analysis of immunophenotype assured accurate lineage diagnosis of AL. Combinatively analyzing the characteristics of AL on morphology, cytochemistry, immunology and MPO mRNA expressions were significant to the diagnosis and therapy of AL.

Objective To explore the significance of dysplasia and cytogenetic changes to the diagnosis and typing of myelodysplastic syndrome (MDS).Methods The dysplasia performance of each series in every isoforms was observed by the bone marrow aspiration and peripheral blood smear to the 132 patients with MDS. At the same time do the chromosome karyotype was analizad combined with morbidness cells and chromosome karyotype abnormal analysis associated with MDS subtype. Resuits Acorrding to the dysplasia ≥0.10, the totle detection rate of granulocyte series, erythrocyte series and megakaryocytic was 43.4 ％.The morbidness granulocyte and megalokaryocyte ≥0.10was mainly in RCMD (P ＜ 0.01); morbidness erythrocytes≥0.10 mainly in RA + RARS (P ＜ 0.01). the totle detection rate of chromosome karyotype abnormal in MDS was 44.0 ％.The detection rate in RA and RARS was lower than other isoforms,but showed no statistically significant (P ＞ 0.05).the relationships of dysplasia and chromosome karyotype abnormal with the isoforms of MDS:in RA group,50.0 ％(3/6) patients had karyotype abnormal simultaneous the detection of morbidness cells≥0.10, 76.0 ％(19/25) in RCMD group and 60.9 ％(14/23) in RAEB group (P ＜ 0.01).Conclusion Theve is relationships between the patients with chromosome karyotype abnormal and dysplasia ≥0.10 and the isoforms of MDS. Closely monitoring the hemopoiesis and cytogenetic changes is significance to diagnose MDS.

Objective To study the recovery of the peripheral lymphocyte subsets in patients underwent allogeneic peripheral blood stem cell transplantation (allo-PBSCT) and guide the prevention and treatment of infection. Methods Indirect immunofluorescence assay was used to detect the lymphocyte subsets, such as T cell subsets (CD3, CD4, CD8). B cell (CD19) and natural killer cell(CD56) at 1, 3, 6, 12, 18months post transplantation, in the meantime, lymphocyte subsets of 32 samples from healthy blood donors were tested as normal control values. Results CD+3, CD+4 and CD+8 ceils significantly decreased than that of normal control at 1 month post transplantation, the recovery of CD+3 T cells was within 3-12 months, CD+4 and CD+8 T cells recovered to normal at 6 months and 3 months post transplantation respectively, CD+4/CD+8 ratio were not significantly lower than that of normal control at different stages, CD+4/CD+8 ratio reversed only at 6 months post transplantation. CD+19 and CD +56 T cells recovered quickly and they were more than normal proportion at 3 months post transplantation. The CD+3, CD+8 T cells and CD+4/CD+8 ratio were statistically higher in HLA haploidentical allo-PBSCT patients than that in HLA identical allo-PBSCT at 3 months post transplantation. There were no difference between the two groups at 1, 6, 12, 18 months post transplantation.The patients with cGVHD had significantly higher CD+4 cells than those without cGVHD at 1 month after transplantation. There was no significant difference in all of the lymphocyte subsets at 3, 6, 12, 18 months after transplantation between them. Conclusion Allo-PBSCT has a hastened immune reconstitution, which was not delayed by the incompatibility of HLA and the development of cGVHD.

Objective To explore the clinical outcome of HLA haploidentical vs HLA-matcbed peripheral blood hematopoietic stem cell transplantation (PBSCT) without in vitro T-cell depletion for malignant hematological diseases. Methods 111 patients with malignant hematological diseases underwent PBSCT without in vitro T-cell depletion between May 2004 and February 2009, including 51 patients with HLA-haploidentical and 60 patients with HLA-matched. All patients have received myeloablative conditioning regimen. A two-agent based graft-versus-host disease (GVHD) prophylaxis was used as cyclosporine A and a short course of methotrexate. Mycophenolate mofetile was added for the patients with one locus mismatch. Mycophenolate mofetile, antithymocyte globulin and CD25 monoclonal antibody were added for the patients with 2-3 loci mismatch. The grafts were granulocyte colony-stimulating factor-mobilized peripheral blood stem cells without in vitro T-cell depletion. Results 111 patients achieved sustained and full donor-type engraftment. The median time to reach an absolute neutrophil count above 0.5×10~9/L was 14 days and that to a platelet count exceeding 20×10~9/L was 15 days in 51 HLA-haploidentical patients, and that was 12 days and 13 days in 60 HLA-matched patients, respectively. In 51 HLA-haploidentical patients, 25 patients developed aGVHD, including 20 cases of grade Ⅰ aGVHD, and 5 cases of grade Ⅱ. Thirty-three patients developed cGVHD with limited in 30 and extensive in 3. The 4-year cumulative incidence of cGVHD was 70.4 %. The 3-year probabilities of leukemia-free survival (LFS) were 74.5% (77.3 % for standard risk patients and 68.2 % for high risk patients respectively). Seven patients had recurrence. In 60 HLA-matched patients, 14 patients developed aGVHD, including 10 cases of grade Ⅰ, 2 cases of grade Ⅱ and 2 cases of grade Ⅲ. Thirty-seven patients developed cGVHD with limited in 32 and extensive in 5. The 4-year cumulative incidence of cGVHD was 58.1%. The 3-year probabilities of LFS were 72.1% (77.6 % for standard risk patients and 52.7 % for high risk patients respectively). Ten patients had recurrence. The incidence of aGVHD in HLA-haploidentical cohort was significantly higher than in HLA-matched cohort (P<0.05). There was no significant difference in incidence of cGVHD, incidence of relapse and LFS between HLA-haploidentical and HLA-matched cohorts (P>0.05). Conclusion Haploidentical PBSCT is feasible and safe for malignant hematological diseases to use myeloablative conditioning regimen in combination with intensive immunosuppressants without in vitro T cell depletion.

BACKGROUND: Lack of human leucocyte antigen-matched family donors has restricted the application of hematopoietic cell transplantation. Due to immunological disorder of humam leucocyte antigen misfit, common way for haploidentical transplantation is associated with poor engraftment and severe graft-versus-host disease. Because not every patient has HLA-Jdentical family member, a substantial proportion of patients will receive haploidentical transplantation. OBJECTIVE: To explore the curative effect on malignant hematological diseases of haploidentical peripheral blood stem cell transplantation (PBSCT) using myeloablative conditioning regimen in combination of proper immunosuppressants without in vitro T-cell depletion. DESIGN, TIME AND SETTING: A case observation was performed at the Department of Hematology in the First Affiliated Hospital of Xinjiang Medical University from July 2002 to June 2008. PARTICIPANTS: Forty-two patients with malignant hematological diseases, including 29 standard-risk patients and 13 high-risk patients, age from 10 to 48 years, were transplanted with cells from a haploidentical family donor with 1-3 mismatched loci of HLA antigens. Seven patients had 1 locus mismatched donors and thirty-five patients had 2-3 loci mismatched donors. METHODS: The patients have received myeloablative conditioning regimen. A two-agent based graft-versus-host disease (GVHD) prophylaxis was used as cyclospodne A and a short course of methotrexate. Mycophenolate mofetile was added for 1 locus mismatched patients. Mycophenolate mofetile, antithymocyte globulin and CD25 mono-colonal antibody were added for 2-3 loci mismatched patients. The grafts were granulocyte colony-stimulating factor-mobilized peripheral blood stem cells without in vitro T-cell depletion. MAIN OUTCOME MEASURES: Engraftment, GVHD incidence and severity, relapse and leukemia-free survival and the immune function of patients in months 1, 3, 6, 12 and 18 postoperatively. RESULTS: Totally 42 patients achieved complete and sustained donor-type engraftment. Nineteen patients developed acute GVHD, the 2-year cumulative incidences of acute GVHD were 50.8%, gradeⅠ acute GVHD occurred in 16 cases and grade Ⅱ in 3 cases. Thirty-one patients were followed up more than 6 months, 23 of them developed chronic GVHD with limited in 20 and extensive in 3, the 2-year cumulative incidences of chronic GVHD were 57.1%. No patients died of GVHD. There were no significant differences in the reduction and recovery of T cells and B cells between HLA haploidentical PBSCT without in vitro T cell depletion and HLA-matched PBSCT. CONCLUSION: Haploidentical PBSCT is feasible and safe for malignant hematological diseases to use myeloablative conditioning regiment combination of intensive immunosuppressants without in vitro T cell depletion. A large amount of clinical cases need to be investigated in the near future.

Xylulose as a metabolic intermediate is the precursor of rare sugars, and its unique pattern of biological activity plays an important role in the fields of food, health, medicine and so on. The aim of this study was to design a new pathway for xylulose synthesis from formaldehyde, which is one of the most simple and basic organic substrate. The pathway was comprised of 3 steps: (1) formaldehyde was converted to glycolaldehyde by benzoylformate decarboxylase mutant BFD-M3 (from Pseudomonas putida); (2) formaldehyde and glycolaldehyde were converted to dihydroxyacetone by BFD-M3 as well; (3) glycolaldehyde and dihydroxyacetone were converted to xylulose by transaldolase mutant TalB-F178Y (from Escherichia coli). By adding formaldehyde (5 g/L), BFD-M3 and TalB-F178Y in one pot, xylulose was produced at a conversion rate of 0.4%. Through optimizing the concentration of formaldehyde, the conversion rate of xylulose was increased to 4.6% (20 g/L formaldehyde), which is 11.5 folds higher than the initial value. In order to further improve the xylulose conversion rate, we employed Scaffold Self-Assembly technique to co-immobilize BFD-M3 and TalB-F178Y. Finally, the xylulose conversion rate reached 14.02%. This study provides a new scheme for the biosynthesis of rare sugars.

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