BACKGROUND: BuCy2 composed of large-dose oral busulfanum and cyclophosphamide has been one of the main conditioning regimens before transplantation, but oral busulfanum can result in many side effects.OBJECTIVE: To evaluate the efficacy and safety of the conditioning regimen with intravenous busulfanum (Ⅳ Bu) in hematopoietic stem cell transplantation (HSCT) for malignant hematopathy.DESIGN: Case observation.SETTING: Department of Hematology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: A total of 13 patients with chronic myelogenous leukemia (CML) and 6 patients with acute leukemia,who received allogeneic HSCT with the conditioning therapy of modified BuCy2, were enrolled at Institute of Hematopathy, Union Hospital, Huazhong University of Science and Technology from May to December 2006. There were 12 males and 7 females aged 14-50 years with an average of 33 years, and there were 9 cases of related transplantation and 10 cases of unrelated transplantation. Hematopoietic stem cells (HSCs) were harvested from peripheral blood of 18 subjects and bone marrow of 1 subject. All patients and their family members signed the informed consent.METHODS: All patients were treated with allogeneic HSCT with the conditioning therapy of intravenous BuCy2.All patients received the modified combination of busulfanum and cyclophosphamide that was commonly utilized currently.Hydroxycarbamide 40 mg/kg was orally taken ten days before transplantation. Arabinosylcytosin (Ara-C) 2 g/m2 was injected via vein nine days before transplantation. Busulfanum ampule 0.8 mg/kg (equal to 1 mg/kg oral administration)was given by central venous cannula eight, seven and six days before transplantation. The injection was performed over 2 hours controlled by infusion pump, once every 6 hours, totally 12 times. Busulfanum was diluted by saline or 5% glucose to about 0.5 g/L (about 10 times) before Ⅳ Bu. Cyclophosphamide 1.8 g/m2 was intravenously injected every day from five and four days before transplantation. Methyl-CCNU 250 mg/m2 was taken orally three days before transplantation. Occurrence of hepatic vano-occlusive disease (HVOD) and its side effects were determined. Follow-up was performed at month 6.5 after operation to observe disease recurrence and patient survival.MAIN OUTCOME MEASURES: Occurrence and side effects of HVOD 100 days after transplantation, and results of follow-up.RESULTS: Totally 19 patients were involved in the result analysis. In 13 days after transplantation, blood conversion,chromosome karyotype and DNA polymorphism tests verified that reconstruction of hematopoietic function was obtained and two patients got recurrence, of which one patient got complete remission after receiving fludarabine plus cytarabine plus granulocyte colony-stimulating factor (G-CSF) and HSCs from peripheral blood. DNA polymorhism test verified that re-engraftment was obtained. Another patient received chemotherapy. Other patients lived well.

Objective To investigate the mechanism of radiosensitizing effects of endostatin on H-520 human lung squamous cancer cells.Methods H-520 cells was treated with endostatin and/or radiation.Colony-forming assays were used to indicate the radiosensitising effects.Cell cycle distribution and expression of phosphor-p38-MAPK were assayed by FCM,and cyclin D1,cdk2,cdk4 and survivin mRNA leveh were assayed by RT-PCR.Phosphor-Akt was evaluated by Western-blotting.Results Combination of endostatin and irradiation inhibited the proliferation of H-520 cells.According to the colony-forming assays,the D0,Dq,D10 and SF2 values of the combination groups were much lower than those of irradiation groups.The sensitization enhancement ratio(SER)was 1.51.G2/M arrest occurred after 4 Gy irradiation.The gene expression of cyclin D1,cdk2,ckd4 and survivin and phosphor-Akt protein were down-regulated after treatment.The expression of phosphor-p38-MAPK protein was also down-regulated after treatment with 200 μg/ml endostar.Conclusions Endostatin inhibits the growth of H-520 cells and radiosensitizes the cells by induction of G0/G1 arrest,cell apoptosis and down-regulation of gene expression of cyelin D1,cdk2,cdk4 and reduces the phosphorylation of Akt and p38-MAPK.

Objective To evaluate the effect of cytokine-induced killer cells (CIKs) as an adoptive immunotherapy option for treatment of leukemia relapse after allo-hematopoietic stem cell transplantation (allo-HSCT).Methods Two cases of infusion of donor CIKs in patients with leukemia relapse after allo-HSCT were retrospectively analyzed.Patient one relapsed 986 days (+986d) after HLA-matched unrelated donor allo-HSCT.Applications of chemotherapy only resulted in short term remission,but allo-CIKs were successfully expanded from the patient's peripheral blood mononuclear cells of donor origin.Totally five cycles of CIKs infusion were infused as an alternative of adoptive immunotherapy.Patient two had recurrent in the + 158d after HLA-matched sibling alloHSCT.At + 204d and + 294d,two cycles of CIKs which were expanded from donor peripheral blood mononuclear cells were infused.Results One cycle of CIKs was given to patient one after the application of chemotherapy to reduce the tumor burden,and the patient successively achieved complete remission.Again after additional four cycles of CIKs infusion,consistent remission was maintained during the following seven months.Patient two who had relapsed disease posttransplantation,achieved cytological complete remission after withdrawal of immunosuppressants and undergoing chemotherapy combined with G-CSF mobilized stem cell infusion.However,at + 187d,the patient suffered from side-effect of acute graft versus host disease and extramedullary infiltration.The symptoms were alleviated markedly after one cycle of CIKs infusion at + 204d.Moreover,the pain disappeared after an additional infusion at + 294d.And up to the present,the bone marrow aspiration showed complete remission while the extramedullary disease vanished.Conclusion The use of CIKs in the treatment of leukemia relapse after allogeneic bone marrow transplantation can be feasible and well tolerated.

Objective To study the relationships between hepatocellular carcinoma (HCC) and the polymorphisms,promoter methylation,and expression of glutathione S-transferases P1 gene (GST) P1 gene.Methods Using methylation-special PCR (MSP),the methylated status of CpG islands of GSTP1 gene in tumor tissues of 53 HCC and its adjacent nontumor tissues were studied.The enzyme activities of GSTP1 were evaluated by ultraviolet colormetry.And using PCR-RFLP,the genetic polymorphisms of the GSTP1 genes of 74 healthy controls and 53 HCC patients were studied.Results The diffe-rences of the frequency of GSTP1 Ile/Ile,Ile/Val and Val/Val genotypes between HCC patients and the normal controls did not reach statistical significance (X~2=0.84,v=2,P=0.656).The frequency of methyla- tion of CpG islands of GSTP1 gene was significantly higher among the HCC tumor tissues when com- pared to the corresponding nontumor tissues (X~2=19.08,P＜0.001),and significantly higher in stageⅢ-Ⅳcases when compared to the stageⅠ-Ⅱcases (X~2=4.84,P=0.028).GSTP1 enzyme activities of cytoplasm in tumor cells were lower significantly than that in the adjacent nontumor tissues (t=2.49, P=0.014),and significantly higher in stageⅠ-Ⅱcases when compared to the stageⅢ-Ⅳcases (t= 2.31,P=0.025).On the other hand,the GSTP1 enzyme activities of cytoplasm in tumor cells with methylated status of GSTP1 gene were significantly lower than that in tumor cells with unmethylation (t=3.50,P=0.001).Conclusion GSTP1 inactivation via CpG island hypermethylation may contrib- ute to the pathogenesis of HCC.

Objective To observe the effectiveness of Antithymocyte globulin/antilymphocyte globulin(ATG/ALG)in the treatment of nonsevere aplastie anemia(NSAA)patients who didn't respond to cyclosporine A(CsA)or relapsed and review the literature.Methods We administered ATG/ALG to 5 NSAA patients who relapsed or showed no re- sponse after CsA therapy.Results 3 patients were cured,2 patients improved,4 patients became transfusion inde- pendent in 90 days,none relapsed or progressed to clonal disease.Conclusion Those NSAA patients who relapsed or showed no response after CsA therapy should be asministered ATG/ALG,so as to reduce transfusion complications and prevent sustained immune attack to the bone marrow.

We used 4 methods,such as ultrasonic crush(UC),ultrasonic rinse(UR),whorl surge(WS)and rubbing(RU),to isolate epiphytic bacteria from red alga Gracilaria lemaneiformis.Then,we counted bacteria numbers,detected bacterial species,observed bacterial configuration and characteristic of cell wall.Compared with these methods and with different treatments in one method,the results were drawn:the UR and RU were inferior in all methods to isolate bacterial numbers and species,the UC and WS were better,especially,the treatment 30W 30s of UC was the best in experiment,which isolated 12 of 16 bacterial species,and got 1.75 10~6 cells per gram wet weight G.lemaneiformis.

Adolescent ; Adult ; Aged ; Humans ; Middle Aged ; Pupil Disorders ; diagnosis ; etiology ; Refraction, Ocular ; Retina ; physiopathology ; Retinal Detachment ; surgery ; Scleral Buckling ; Visual Acuity ; Vitreoretinopathy, Proliferative ; surgery

BACKGROUND: Allogeneic hematopoietic stem cell transplantation is recognized as the only method of curing chronic myelocytic leukemia (CML). Lmatinib mesylate (STI571) is a competitive inhibitor of the bcr-abl tyrosine kinase, as a represent of synthetic gene-targeting drug in recently, which has been used more and more on the Philadelphia chromosome positive CML patients.OBJECTIVE: To compare the efficacy and safety of STI571 to related allogenic hematopoietic stem cell transplantation in the treatment of CML patients.DESIGN, TIME AND SETTING: A controlled observation between ST1571 treated group and transplantation group was performed in the Department of Hematology, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology between April 2002 and October 2006. PARTICIPANTS: All 90 patients with CML in the chronic phase were selected from Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, and they were diagnosis based on the examinations of bone marrow morphologic, cytogenetics and/or molecular genetics. METHODS: All 90 patients with CML in the chronic phase were divided into two groups. 67 patients received oral STI571 (400 mg/day) in succession at the beginning time from April 2002 to June 2006, and the observation ended until October 2006, Blood routine will be done weekly, and bone marrow morphologic and cytogenetic examination would be done every three months. Other 23 patients selected from Union Hospital from March 1999 to April 2006 accepted allo-HSCT, with BuCy2 or modified BuCy2 as conditioning regimens. Cyclosporin A combining with short-term MTX were used in all patients for prophylaxis of graft-versus-host disease (GVHD). MAIN OUTCOME MEASURES: Hematology responses, eytogenetic response and two years survival in two groups were observed. RESULTS: Complete cytogenetic response was achieved in 60% and 100% of the patient treated with STI571 and transplantation respectively (P < 0.01). But two years survival of ST1571 and transplantation were 83.33% and 77.03% respectively, and no difference was found between the two groups (P > 0.05). No one died or discontinued therapy for adverse effects, and 4 out of 67 (5.97%) had grade 3 or 4 thrombocytopenia and/or leucopenia in the ST1571 group. Moreover, in transplantation group, 7 patients (30.4%) developed grade 2 to 4 acute GVHD, but 4 died of failed treatment. CONCLUTION: Compared with transplantation, patients treated with ST1571 achieved low complete cytogenetic responses and the treatment-related complications were mild and manageable or no need for treatment.

ObjectiveTo evaluate the primary effect of granulocyte-monocyte colony stimulating factor (GM-CSF) as an immunotherapy option for treatment of residual disease after alloHSCT.Methods Immunotherapy was performed on two patients with blood malignancy to treat residual disease after allo-HSCT. The patient one,who was diagnosed as having MDS-RAEB Ⅱ,showed bone marrow displasis and incomplete chimerism 6 months after unrelated donor HSCT.Immunosuppressive drug was withdrawn without induction of graft-versus-host disease (GVHD).The patient two B-ALL demonstrated a residual disease at molecular level 30 days post-transplantation.Both of them were given GMCSF (300 μg) subcutaneously once every two days for totally three weeks.During the whole period,skin itch and rash,liver function,subgroups of lymphocytes,and MDSCs and DCs in peripheral blood were investigated.Results In case one,grade Ⅰskin acute GVHD (aGVHD) appeared as early as one week after GM-CSF administration,as well as grade Ⅱ (skin and liver) by the end of the third weeks,and GM-CSF injection was withdrawn.One month later since the start of GM-CSF,the patient showed normal bone marrow morphology and full donor type chimerism. Cyclosporine A (CsA), mycophenolate mofetil and methylprednisolone were administered for two weeks to control GVHD.In the other case,grade Ⅰ aGVHD occurred 9 days after GMCSF administration,and whole blood CsA maintained at 0.134-0.472 μmol/L.Prednisone (30mg per day for 5 days) was used to control grade Ⅱ GVHD from the 11th day after GM-CSF,and grade Ⅰ GVHD continued without any intervention.On the 30th day after GM-CSF treatment,bone marrow aspiration showed complete molecular remission.In both of the two cases,no differences in lymphocytic subtypes were revealed before and after GM-CSF administration,while there were trends of increased DC number and decreased MDSCs in peripheral blood.ConclusionThe administration of GM-CSF as an immunotherapy option for blood malignancy may contribute to the clearance of residual disease after Allo-HSCT.