Objective: To explore the effect of simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (the rate-limiting enzyme of cholesterol synthesis) on the proliferation and apoptosis of chronic myelogenous leukemia (CML) cells. Methods: Both normal and CML bone marrow progenitor cells were assayed in semisolid methylcellulose culture after incubation for 24 hours in suspension culture with 10 mg*L－1 simvastatin. Also, sub-G1 cells and DNA end-labeling positive cells as apoptotic cells were identified by flow cytometry after being exposed to simvastatin for 72 hours. Results: Simvastatin selectively inhibited proliferation and induced apoptosis of CML cells, but had no much influence on normal bone marrow cells. Conclusion: CML cells are more sensitive to a short-term exposure to simvastatin than normal bone marrow cells. It will be a promisingly effective chemotherapeutic agent or in vitro purging agent in autologous stem cell transplantation for the treatment of CML.

Objective: To investigate hyperthermia enhanced expression of heat shock protein70 (HSP70) in breast cancer drug sensitive cell line MCF7 and multi-drug resistant (MDR) cell line MCF7/ADR, so as to increase cytotoxic activity of immunologic effector cells against the target cells, and to provide an experimental basis for cell immunotherapy based on hyperthermia. Methods: The immunological effector cells were induced and expanded by cytokines. Breast cancer cell lines MCF7 and MCF7/ADR were treated at 42 ℃ for an hour. Then after being incubated for additional 4 hours, 24 hours and 48 hours, the cells were digested. Flow cytometry (FCM) was used to detect the expression of HSP70 on the target cells. MTT assay was employed to evaluate cell proliferation and the cytotoxic activity of immunologic effector cells against target cells. Results: The expressions of HSP70 in both the target cells had significant difference. The expression of HSP70 in MCF7/ADR cells was higher than in MCF7 cells before hyperthermia. After hyperthermia the expression of HSP70 increased by 6 folds in MCF7/ADR cells, and 22 folds in MCF7 cells and was higher than in MCF7/ADR cells at hour 4. The proliferation inhibition fraction of hyperthermia against MCF7 was significantly lower than that of MCF7/ADR. The cytotoxic activity of immunologic effector cells against MCF7 cells was lower than against MCF7/ADR cells before hyperthermia. After hyperthermia the cytotoxic activity of immunologic effector cells against MCF7 cells rose by 86.23%, and was higher than against MCF7/ADR cells (rose by 30.32%). Conclusion: The expression of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR were enhanced significantly by hyperthermia. Cytotoxic activity of immunologic effector cells against both the target cells were increased by hyperthermia. The differential expressions of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR affect the cytotoxic activity of immunologic effector cells.

Objective To assess the effect of low-dose cytarabine and aclarubicin in combination with gran-ulocyte colony-stimulating factor (G-CSF) protocol (CAG) in patients with acute myeloid leukemia (AML),and to understand the potential factors affecting the outcome of CAG induction therapy, therefore to find the optimum pa-tients for CAG therapy. Methods Twenty-one AML patients were enrolled in the current study. All patients were treated with CAG regimen including cytarabine (10 mg/m~2, subcutaneously, every 12 h, days 1 - 14), lacinomycin (5～7 mg/m~2,intravenously,every day, days 1 -8) ,and G-CSF (200 μg/m~2,subcutaneously, every day,12 h be-fore Ara-C was given) priming. Results The overall complete remission (CR) rate of the 21 AML patients was 66.7% (14/21). The CR rates was 87.5% (7/8) in patients older than 60 yrs,60.0% (9/15) in the refractory or relapsed patients,83.3% (5/6) in the MDS transformed AML patients. The CR rates for patients with hyperprolif-erative BM and median to poor proliferative BM were 33.3% and 91.7% ,respectively(P =0.009). The median o-verall survival (OS) time of the 21 AML patients was 450 days. Two-year survival rate estimated by Kaplan-Meier Method was 30.6%. The overall median disease free survival (DFS) was 165 days. The median OS time for those refractory or relapsed was 435 days. The median OS time for those with poor cytogenetic state or standard or good cytogenetic state was 140 days and 620 days, respectively (P = 0.001). The median OS time for patients with hyperproliferative BM and median to poor proliferative BM was 321 days and 620 days, respectively (P = 0.05). The median recovery time of granulocytes above 1.0×10~9/L was 8 days. The median duration of fever was 3.5 days. The rate of infections exceeding WHO grade Ⅱ was 42.9%. No early death occurred. Conclusions The CAG induction therapy may have a higher CR rate in patients with refractory or relapsed AML, elderly AML and secondary AML from MDS transformation, and extend the median overall survival time in refractory or relapsed patients. CAG therapy can not improve the outcome of patients whose BM was in high grade proliferation state or whose cytogenetic state was poor. CAG therapy can shorten the duration of agranulocytosis and decrease the inci-dence of serious infection. Therefore, CAG therapy is worth recommending to patients who can not endure the rou-tine intensive chemotherapy.

Objectives To explore the anti-apoptotic mechanism and the effective apoptosis-inducing method in chronic myelogenous leukemia (CML) cells.Methods K562 cell line was used to observe the effect of combination of herbimycin A (HMA), a tyrosine kinase inhibitor, and chemotherapeutic agents on the induction of apoptosis.Results HMA or chemotherapeutic agents could inhibit the proliferation but not significantly induce apoptosis of K562 cells. However, HMA significantly enhanced apoptosis when combined with chemotherapeutic agents. Addition of sulfhydryl compound to the cultures to conjugate HMA completely abrogated this enhancing effect on K562 cells.Conclusions HMA increases the sensitivity of CML cells to chemotherapeutic agents by inactivating tyrosine kinase activity. It is promising that combination of HMA with conventional chemotherapeutic drugs may be a new strategy in the treatment of CML.

OBJECTIVETo investigate the application of allogeneic peripheral blood stem cell transplantation (allo-PBSCT) in the treatment of hematologic malignancies.

METHODSBetween October 1995 and August 2001, fifty-one patients with hematologic malignancies (median age 34 years, range 5.5 approximately 52 years) received allo-PBSCT from HLA-identical (50) or 1-antigen mismatched sibling donors with conditioning regimens of TBI + CY or modified BU/CY2. Thirty-one patients were acute leukemia (AL) (15 in CR(1), 7 in CR(2) or greater, 10 in relapse including 2 relapse after allo-BMT and the other one never achieved remission); 12 chronic myeloid leukemia (CML) (CP 5, AP 2, BC 4 and relapse after allo-BMT 1); 7 MDS (RAEB 1, RAEB-T 1, AL secondary to MDS 5); Burkitt's lymphoma 1. A combination of cyclosporine and methotrexate was administered for GVHD prophylaxis.

RESULTAll patients were engrafted. The median time (range) to neutrophil >/= 0.5 x 10(9)/L and platelet >/= 20 x 10(9)/L was 14 (10 approximately 20) and 11 (7 approximately 45) days post-transplant, respectively. Grade II approximately IV acute GVHD occurred in 20/51 (39%) and grade III approximately IV aGVHD in 2 patients. Clinical chronic GVHD was diagnosed in 23 of 44 (52%) evaluable patients. Fourteen patients died: 8 died of transplant related complications, 6 of relapse. Thirty-seven patients are alive with a median follow-up of 399 (75 approximately 2 176) days, and among them 34 are in continuous complete remission, the other 3 relapsed. The 2-year probability of overall survival, disease-free survival (DFS) and relapse is 64%, 61% and 24%, respectively.

CONCLUSIONAllogeneic PBSCT is safe for both donors and recipients, and results in a rapid and stable engraftment without increase in incidence or severity of acute GVHD.

Acute Disease ; Adolescent ; Adult ; Child ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Hematologic Neoplasms ; mortality ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Survival Analysis ; Survival Rate ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo observe the antileukemic effect in relapse patients by infusion of donor immunocompetent cells with or without granulocyte colony-stimulating factor (G-CSF) mobilization.

METHODSTwenty patients with leukemia in relapse after allogeneic bone marrow transplantation (allo-BMT) were treated with chemotherapy followed by donor-derived lymphocytes (DDL) without G-CSF mobilization (Group A, n = 11), or donor peripheral blood progenitor cells (PBPCs) with G-CSF mobilization (Group B, n = 9).

RESULTSFive patients in Group A were in hematologic relapse. After DDL infusion, 3 of 5 patients had a temporary complete remission (CR) and relapsed after 3, 7 and 10 months, respectively. One achieved partial remission and died of interstitial pneumonia; and the other one showed no response. Another 6 patients in Group A were in cytogenetic relapse or central nerve system (CNS) leukemia, and all achieved CR and remained in disease free survival (DFS) for 10 to 98 months after DDL infusion. All 9 patients in group B were in hematologic relapse. Three patients with chronic myeloid leukemia (CML) had cytogenetic and molecular remission for 16, 35 and 51 months, respectively after PBPC infusion; and 5 patients with acute lymphoid leukemia (ALL) had CR and were still in CR for 10 to 18 months except 1 patient relapsed soon. And the other one with AML showed no response to the therapy.

CONCLUSIONDonor immunocompetent cells infusion is an effective therapy for relapsed leukemia after allo-BMT, especially for the patients with early (molecular and cytogenetic) or CNS relapse. Infusion of donor PBPC mobilized by G-CSF seems to have more potentiated graft-versus-leukemia (GVL) effect than DDL infusion.

Adolescent ; Adult ; Bone Marrow Transplantation ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; therapy ; Male ; Middle Aged ; Recurrence

Objective: To investigate the therapeutic effects of allogeneic hematopoietic stem cell transplantation (allo-HSCT) with FLAG sequential busulfan/cyclophosphamide(Bu/Cy) conditioning regimen for refractory/relapsed acute myeloid leukemia. Methods: From February 2012 to June 2017, 21 patients with refractory/relapsed acute myeloid leukemia underwent allo-HSCT with FLAG sequential Bu/Cy conditioning regimen. Transplantation-related complications and clinical outcome were retrospectively analyzed. Results: After conditioning, no hepatic veno-occlusive disease (VOD) and grade Ⅲ hemorrhagic cystitis occurred. 76.2% (16/21) patients had fever with 4 septicemia. One patient died of septic shock before engraftment. Twenty patients achieved neutrophil engraftment with a median time of 13 days (range, 10 to 21 days). Seventeen patients achieved platelet engraftment with a median time of 18 days (range, 9 to 25 days). The cumulative incidence of acute graft-versus-host disease (aGVHD) was 39.5%, and 3 patients developed grade Ⅲ-Ⅳ aGVHD. Of 19 patients who survived more than 100 days after transplantation, 4 had local chronic graft-versus-host disease (cGVHD). Of 21 patients, the median survival time was 15 months (range, 0.5 to 67 months) post-transplantation. Transplantation-related mortality rate was 28.7%. Leukemia relapse occurred in 4 patients with a median time of 4 months (range, 3 to 8 months) after transplantation. The cumulative relapse rate at 1 year was 21.4%. The 1-year and 3-year overall survival (OS) rates were 60.7% and 54.9% respectively. Log-rank analysis revealed that bone marrow blasts ≥ 20% or extramedullary leukemia before transplantation, poor platelet engraftment and grade Ⅲ-Ⅳ aGVHD were significantly related to shortened OS (P<0.05). Conclusions Allo-HSCT with FLAG sequential Bu/Cy conditioning regimen in patients with refractory/relapsed myeloid leukemia has acceptable transplantation-related risk and relapse rate. The 1-year and 3-year OS rates are comparable with those in remission patients.

Objective:To investigate the clinical features,therapeutic methods,therapeutic efficacy,and prognostic characteristics of older patients with acute myeloid leukemia(AML).Methods:We collected data from 134 older patients with AML treated at Peking University International Hospital between January 2015 and February 2023.White blood cell count,bone marrow primitive cell count,cytogen-etic and molecular characteristics,and European LeukemiaNet(ELN)risk stratification at initial diagnosis were retrospectively ana-lyzed.Patients were assigned into two groups according to treatment plan―high-intensity chemotherapy and low-dose treatment―to determine whether intensive chemotherapy would yield survival benefits during treatment and the factors affecting survival.Results:Among 36 patients treated with high-intensity chemotherapy,22(61.1%)achieved complete response(CR);among 90 treated with low-intensity therapy,46(51.1%)achieved CR;and among 19 treated with azacitidine(AZA)+ venecra(VEN),14(73.7%)achieved CR.Medi-an overall survival(OS)was 15 months for high-intensity chemotherapy and 14.5 months for low-intensity treatment(P=0.226).According to ELN risk stratification,patients in the low,medium,and high risk groups exhibited OS of 18,14,and 9 months,respectively(P=0.009).OS for high-intensity chemotherapy and low-dose therapy was 22 and 15 months in the low-risk group(P=0.745),9 and 15 months in the medium-risk group(P=0.783),and 9 and 8 months in the high-risk group(P=0.739),respectively.Patients in the intensive chemotherapy group(n=36)had an OS of 15 and 17 months(P=0.689)compared with AZA+VEN treatment(n=19).The prognosis of six patients with TP53 mutation was significantly worse than those without the mutation,and the median OS was 2 months and 14 months,respectively(P=0.004).One-and 3-year survival rates for the low-,medium-and high-risk groups were 79%,53%,and 44%,and 41%,20%,and 3%,respectively.Multivariate analysis revealed that high peripheral blood white blood cell count(P=0.034),ELN risk stratification(P=0.002),and complications(P=0.017)were correlated with OS,while treatment intensity,age,sex,and bone marrow primitive cell count were not significantly correlated with OS.Conclusions:High-intensity chemotherapy did not yield a significant survival benefit in older patients with AML;however,this result needs to be confirmed in patients at low risk.Patients with TP53 mutations had a poor prognosis.Multivariate analyses revealed that baseline mo-lecular characteristics,leukocyte count,and comorbidities were more important than treatment intensity in predicting survival among older patients with AML.

Objectives:To investigate the diagnostic value of whole blood quantitative PCR for DNA load of Epstein-Barr virus (EBV) in post-transplant lymphoproliferative disease (PTLD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) .Methods:A total of 694 patients with hematologic diseases who underwent allo-HSCT at the Hematology Department of Peking University First Hospital from April 2004 to April 2019 were included, and their data were retrospectively analyzed.Results:①Among the 694 cases, 29 cases (22 males and 7 females, with a median age of 22 (1-52) years) developed PTLD after allo-HSCT with a cumulative incidence of 4.2% and a median onset time of 2.1 (0.8-20.6) months. ② Univariate analysis showed that age<30 years, diagnosis with aplastic anemia, human leukocyte antigen (HLA) mismatch, use of antithymocyte globulin (ATG) in preconditioning regimens, and EBV reactivation were the risk factors for the occurrence of PTLD. Multivariate analysis showed that EBV reactivation was an independent risk factor for the occurrence of PTLD. ③Further analysis of EBV reactivation cases showed that the peak value of EBV-DNA load was significantly higher in the PTLD group than that in the non-PTLD group ( P<0.001) and the incidence of PTLD increased with the increase of EBV-DNA load. Receiver operating characteristic (ROC) curve analysis indicated that PTLD was more likely to be diagnosed when the EBV-DNA load was >1.19×10 6 copies/ml (sensitivity 0.800 and specificity 0.768) . ④All patients with PTLD received rituximab-based treatment, with an overall response rate of 86.2% and an overall survival rate of 54.3%. Conclusion:The PTLD occurrence after allo-HSCT is highly correlated with EBV reactivation, and the higher the EBV-DNA load, the greater the risk of PTLD occurrence. The dynamic monitoring of EBV-DNA load plays an important role in predicting PTLD occurrence.