Objective To study the clinical outcome of patients with fns-like tyrosine kinase-3internal tandem duplication (FLT3-ITD) positive acute myeloid leukemia (AML) treated with allogeneic hematopoietic stem cell transplantation (allo-HSCT) and to explore the potential prognostic factors to patients' survival including transplant types or disease status.Methods A total of 314 AML patients in our center from October 2006 to October 2012 were retrospectively analyzed,among whom,54 patients were defined with FLT3-ITD positive.Survival rates and treatment-related mortality were further analyzed.Results For all 54 FLT3-ITD positive patients,the 3-year overall survival (3-OS) rate was 56％ and 3-year leukemia-free survival (3-LFS) rate was 47％.The outcome of haplo-identical HSCT was similar as that of sibling donors(3-OS rate:60％ vs 54％ ; 3-LFS rate:54％ vs 45％,respectively).There were 47 patients who received transplantation in first complete remission(CR1).The other 7 patients were of disease relapse or in CR2 before transplantation.Not surprisingly,patients in CR1 had better prognosis than those in nonCR1.Conclusions Allo-HSCT is an effective treatment for AML patients with FLT3-ITD positive mutation.The survival outcome of haplo-identical HSCT was comparable with that of sibling donors.Relapse of AML was the dominant factor related to the mortality of FLT3-ITD positive AML patients after allo-HSCT.

Objective To explore the influence of the killer cell immunoglobulin like receptor (KIR) gene polymorphism on cytomegalovirus (CMV) infection and pathogenesis after hematopoietic stem cell transplantation (HSCT).Methods The KIR genotype was determined by sequence-specific primer polymerase chain reaction (PCR-SSP) in 138 pairs of donors and recipients before HSCT during October,2005 and May,2011.Posttransplant monitoring for CMVpp65 antigen was performed by indirect immune histochemically assays since week 2 after transplantation.The differences between CMV positive group and negative group,inhibitive and active KIR of donors and recipients,and KIR haplotype frequency of donors and recipients were analyzed.Results There were no significant differences in frequency of KIR gene and haplotype AA,AB,BB between the donors and recipients.The frequencies of 2DS2 and 2DS4 * 003-007 of donors in CMV positive group were obviously lower than those in CMV negative group with significant differences(8％ vs 16％,P =0.0420;3％ vs 13％,P =0.0050).There was no significant difference in KIR gene between CMV positive group and CMV negative group.The CMV infection rates of haplotype AA,BB,AB donors were 64.38％,36.84％ and 50.00％,while CMV infection rates of haplotype AA,BB,AB recipients were 53.73％,46.15％ and 51.72％,respectively.The CMV infection rate was higher in the patients received KIR haplotype AA donor than in those received KIR haplotype BB donor (36.84％ vs 64.38％,P =0.0299).2DS4 * 003-007 and haplotype BB of donor were found associated with CMV infection in multifactor analysis.Conclusion KIR genotypes of donors are associated with CMV infection after HSCT.

Objective:To investigate the incidence, risk factors and survival of cGVHD patients in combination of a haploidentical donor supported with an unrelated umbilical cord blood for hematopoietic stem cells transplantation (haplo-cord-HSCT).Methods:300 hematological malignancies individuals who received dual transplantation were enrolled in the study between January 2012 and July 2016 at the department of Hematology in the First Affiliated Hospital of Soochow University. The clinical diagnosis and scoring the severity of cGVHD syndromes according the National Institutes of Health (NIH) consensus conference in the 2014 update. Cox proportional hazards regression was used to identify risk factors associated with transplant outcomes.Results:During follow-up with a median time of 26.4 months (range 0.2-61.8) post transplantation, the 1-year, 3-year and 5-year cumulative incidence of cGVHD was 26.3 % (95 % confidence interval [CI], 23.5 %~29.1 %), 30.3 % (95 % CI, 27.3 %~33.3 % ) and 32.2 % (95 % CI, 28.7 %~35.7 %). For all 73 patients with cGVHD, first-line or second-line treatment were given. During the follow-up period, 53 patients survived, and 20 patients died. In multivariate analysis, the cGVHD overall survival (GOS) were associated with thrombocytopenia(<100×109/L)(HR=0.103, 95 % CI 3 %-36.1 %, P<0.001). Conclusions:Our data suggest that, the 5-year cumulative incidence of cGVHD was 32.2 % after haplo-cord-HSCT with hematological malignancies. Thrombocytopenia (<100×109/L)was independent risk factors for GOS.

OBJECTIVETo clarify the clinical, cytogenetical and molecular characteristics and prognosis of Ph(+) ALL patients with ABL kinase domain mutations (ABL-KDMs), and to evaluate the therapeutic value of allogeneic hematopoietic stem cell transplantation (allo-HSCT) combined with tyrosine kinase inhibitor (TKI) in these patients.

METHODSRetrospective analysis of clinical features, molecular genetic characteristics, mutation distribution and prognosis of newly diagnosed Ph(+) ALL patients with ABL-KDMs from February 2010 to August 2014 were performed, and the efficacy of treatment regimen of allo-HSCT combined with different TKIs was compared.

RESULTSOf 88 Ph(+) ALL patients during maintenance treatment stage for ABL-KDMs monitoring, mutation was detected in 42 patients with median time of 8 months from diagnosis to mutation occurrence. The median age of mutation group was 40-year-old, older than that of non-mutation group (32.5-year-old) (P=0.023). The incidence of complex chromosome abnormality of mutation group was higher than that of non-mutation group (P=0.043), with alternations in chromosome 7, 5 and +Ph more frequently observed. There were 21 types of mutation at 18 locations detected, with T315I mutation ranking the top followed by E255K/V, Y253H/F and E459K. Mutation group featured no significant difference in complete remission (CR) rate in contrast to nonmutation group, but was remarkably lower in major molecular remission (MMR) rate than non-mutation group. The 2 year and 5 year overall survival rate of mutation group was 45.4% and 35.0% respectively, much shorter than that of non-mutation group (67.8% and 63.3%), (P=0.047). The median survival of patients with T315I and E255K/V was 19 and 10 months, significantly shorter than that of patients with other mutations. Among the 42 patients with mutations, 14 underwent allo-HSCT, and the median survival was 29 months, longer than that of patients received chemotherapy alone (17 months) (P=0.024). Fourteen allo-HSCT patients were given nilotinib or dasatinib at the time of mutation occurrence, and there was no significant difference in the overall survival in contrast to patients who continue to take imatinib.

CONCLUSIONSABL kinase domain mutations are closely related to the older age and high genomic instability in the newly diagnosed Ph(+) ALL patients. Mutation types showed diversity and complexity, which remarkably affected patients' prognosis and survival. T315I and E255K mutations account for more than half of all cases, characterized by a less favorable prognosis. Currently, allo-HSCT is the only method that has the potential of elongating life expectancy, but the utility of second-generation TKI during relapse does not necessarily have an edge on survival over imatinib.

Chromosome Aberrations ; Dasatinib ; therapeutic use ; Hematopoietic Stem Cell Transplantation ; Humans ; Imatinib Mesylate ; therapeutic use ; Mutation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; Prognosis ; Protein Kinase Inhibitors ; therapeutic use ; Proto-Oncogene Proteins c-abl ; genetics ; Pyrimidines ; therapeutic use ; Remission Induction ; Retrospective Studies ; Survival Rate

Objective To investigate the monitoring significance of WT1 gene level in the prognosis of acute myeloid leukemia (AML) patients with normal karyotype after hematopoietic stem cell transplantation (HSCT). Methods The clinical data of 115 AML patients with normal karyotype who were treated with HSCT from July 2009 to March 2017 in the First Affiliated Hospital of Soochow University were retrospectively analyzed. The dynamic detection of bone marrow WT1 gene was carried out by using reverse transcription_polymerase chain reaction (RT_PCR). According to the relative expression level median of WT1 gene before transplantation, the whole patients were divided into the two groups (

Object To examine the preliminary clinical efficacy of custom-made three-dimensional(3D) printed talus prosthesis in the treatment of collapse talus necrosis. Methods:The clinical data of 8 patients who received 3D printed custom-made talus prostheses replacement for severe collapsed necrosis of the talus at the Orthopaedic Sports Medical Center, the First Affiliated Hospital to Army Medical University were analyzed retrospectively.All patients were male,with an average age of 38.0 years (range:22 to 65 years).There were 5 cases of left talus collapse and 3 cases of right talus collapse,with the course of disease of 29.7 weeks (range:6 to 96 weeks).The CT data of contralateral healthy talus were used for mirror image design references for the prosthesis,and the electron-beam 3D printing technology was used to prepare the prosthesis.Titanium alloy (Ti6Al4V) was taken as the material for the preparation of the talus body prosthesis,and Co-Cr-Mo material was used as the material for the preparation of the tibialis talus lateral joint surface prosthesis,and the subtalar joint surface of the prosthesis was made from a microporous casting technique.The prosthesis was analyzed preoperatively by digital three-dimensional finite element analysis and solid comparison techniques to measure anatomic match of the prosthesis.A longitudinal incision on medial ankle was made.The necrotic talus was completely removed and the prosthesis was then implanted.The patient was reexamined in the outpatient department 3, 6, and 12 months after surgery.Primary outcome measures were the American Orthopaedic Foot and Ankle Society(AOFAS) ankle-hind foot score,visual analogue scale(VAS) and ankle range of motion.Changes in imaging data and plantar pressure were also assessed.Repeated measures analysis of variance and paired- t test were used to compare the data. Results:The talus prosthesis measure preoperatively was completely consistent with that contralateral healthy talus and there was no operation-related complication. All the wounds healed primarily. The patients were followed up effectively for 23.17 months (range:12 to 48 months).The preoperative dorsiflexion of patients was (7.6±5.7)°,it increased to(14.2±6.6)° at 12 month after surgery ( t=-2.67, P=0.03).The plantar flexion increased from (22.0±9.9)°preoperatively to (29.2±8.7)° at 12 month after surgery ( t=-8.95, P<0.01).Preoperative AOFAS ankle-hind foot score was 26.3±6.6,and it increased to 70.1±2.2,76.0±3.4 and 79.3±4.2 at 3 month,6 month and 12 month after surgery( F=56.81, P<0.01);Pre-operative VAS was[ M( Q R)]3.0(0.8),and it increased to 2.5(1.0),1.5(1.0),1.0(1.0)at 3 month,6 month and 12 month after surgery( F=20.00, P<0.01).At the last follow-up,imaging reexamination showed that the prosthesis of all patients were in stable position with no sign of subsidence.No secondary ankle fusion or revision was required.The talus height increased from (27.6±6.0)mm preoperatively to (34.6±3.5)mm ( t=-2.94, P<0.01).The plantar pressure showed that the maximum pressure on the healthy ankle was(629.9±26.1)N,and that on the affected side was(521.4±14.4)N.The pressure on the healthy ankle was(350.6±29.6)N,and that on the necrotic side was (212.3±9.7)N.The load on the contralateral forefoot was(38.1±2.8)% and that on the necrotic side was(11.5±2.0)%.The load on the contralateral hindfoot was (24.6±2.5)% and that on the necrotic side was (21.1±1.8)%. Conclusions:The custom-made 3D printed talus prosthesis could restore the talus anatomy,recover the ankle joint function,relieve the pain of patients and improve the life quality of patients.The effect on plantar pressure is mainly achieved by adjusting the center of gravity of plantar pressure backwards and the increase of weight bearing of the healthy foot.

Object To examine the preliminary clinical efficacy of custom-made three-dimensional(3D) printed talus prosthesis in the treatment of collapse talus necrosis. Methods:The clinical data of 8 patients who received 3D printed custom-made talus prostheses replacement for severe collapsed necrosis of the talus at the Orthopaedic Sports Medical Center, the First Affiliated Hospital to Army Medical University were analyzed retrospectively.All patients were male,with an average age of 38.0 years (range:22 to 65 years).There were 5 cases of left talus collapse and 3 cases of right talus collapse,with the course of disease of 29.7 weeks (range:6 to 96 weeks).The CT data of contralateral healthy talus were used for mirror image design references for the prosthesis,and the electron-beam 3D printing technology was used to prepare the prosthesis.Titanium alloy (Ti6Al4V) was taken as the material for the preparation of the talus body prosthesis,and Co-Cr-Mo material was used as the material for the preparation of the tibialis talus lateral joint surface prosthesis,and the subtalar joint surface of the prosthesis was made from a microporous casting technique.The prosthesis was analyzed preoperatively by digital three-dimensional finite element analysis and solid comparison techniques to measure anatomic match of the prosthesis.A longitudinal incision on medial ankle was made.The necrotic talus was completely removed and the prosthesis was then implanted.The patient was reexamined in the outpatient department 3, 6, and 12 months after surgery.Primary outcome measures were the American Orthopaedic Foot and Ankle Society(AOFAS) ankle-hind foot score,visual analogue scale(VAS) and ankle range of motion.Changes in imaging data and plantar pressure were also assessed.Repeated measures analysis of variance and paired- t test were used to compare the data. Results:The talus prosthesis measure preoperatively was completely consistent with that contralateral healthy talus and there was no operation-related complication. All the wounds healed primarily. The patients were followed up effectively for 23.17 months (range:12 to 48 months).The preoperative dorsiflexion of patients was (7.6±5.7)°,it increased to(14.2±6.6)° at 12 month after surgery ( t=-2.67, P=0.03).The plantar flexion increased from (22.0±9.9)°preoperatively to (29.2±8.7)° at 12 month after surgery ( t=-8.95, P<0.01).Preoperative AOFAS ankle-hind foot score was 26.3±6.6,and it increased to 70.1±2.2,76.0±3.4 and 79.3±4.2 at 3 month,6 month and 12 month after surgery( F=56.81, P<0.01);Pre-operative VAS was[ M( Q R)]3.0(0.8),and it increased to 2.5(1.0),1.5(1.0),1.0(1.0)at 3 month,6 month and 12 month after surgery( F=20.00, P<0.01).At the last follow-up,imaging reexamination showed that the prosthesis of all patients were in stable position with no sign of subsidence.No secondary ankle fusion or revision was required.The talus height increased from (27.6±6.0)mm preoperatively to (34.6±3.5)mm ( t=-2.94, P<0.01).The plantar pressure showed that the maximum pressure on the healthy ankle was(629.9±26.1)N,and that on the affected side was(521.4±14.4)N.The pressure on the healthy ankle was(350.6±29.6)N,and that on the necrotic side was (212.3±9.7)N.The load on the contralateral forefoot was(38.1±2.8)% and that on the necrotic side was(11.5±2.0)%.The load on the contralateral hindfoot was (24.6±2.5)% and that on the necrotic side was (21.1±1.8)%. Conclusions:The custom-made 3D printed talus prosthesis could restore the talus anatomy,recover the ankle joint function,relieve the pain of patients and improve the life quality of patients.The effect on plantar pressure is mainly achieved by adjusting the center of gravity of plantar pressure backwards and the increase of weight bearing of the healthy foot.

OBJECTIVETo investigate the effect of nuclear transcription factor RelB on the proteasome inhibitor-sensitivity of chronic lymphocytic leukemia (CLL) cells.

METHODSThe mRNA expression of RelB in CD5⁺ CD19⁺ CLL cells from BM was analyzed by reverse transcription PCR (RT-PCR). The RelB activity was examined by electromobility shift assay (EMSA) and an ELISA-based NF-κB family transcription factor activity assay. CLL cells were classified into RelB+ and RelB- groups according to RelB activity. The frequencies of cell death of CLL cells cultured with human bone marrow stromal cells (hBMSCs) after treatment with PS-341, MG-132 or fludarabine were determined by PI staining.

RESULTSRelB mRNA expression and RelB activity could be detected in CLL cells at variable levels. Fludarabine (10 μmol/L), MG-132 (1 μmol/L) and PS-341 (1 μmol/L) could induce cell death of RelB+ and RelB- CLL cells co-cultured with hBMSCs in a time dependent manner. There was no significant difference in the fludarabine sensitivity between RelB+ and RelB- CLL cells, and the frequencies of cell death of RelB+ and RelB- CLL cells were (61.11 ± 6.91)% and (67.57 ± 9.45)%, respectively, when treated with fludarabine for 72 h. RelB+ CLL cells were more sensitive to MG-132 than RelB- CLL cells for 72 h, and the frequencies of cell death were (66.22 ± 3.39)% and (51.07 ± 5.93)%, respectively. RelB+ CLL cells were more sensitive to PS-341 than RelB- CLL cells for 24 and 48 h treatment, and the frequencies of cell death of RelB+ and RelB- CLL cells were (75.50 ± 4.66)% and (66.32 ± 10.20)% for 24 h, (92.11 ± 3.14)% and (85.84 ± 5.81)% for 48 h treatment, respectively.

CONCLUSIONThe alternative NF-κB activity was detected in bone marrow derived CLL cells. Enhancement of RelB activity may increase CLL cells' sensitivity to proteasome inhibitor bortezomib and MG-132. However, the sensitivity of CLL cells to fludarabine had no relationship to RelB activity.

Cell Death ; drug effects ; Drug Resistance, Neoplasm ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; pathology ; NF-kappa B ; metabolism ; Proteasome Inhibitors ; pharmacology ; RNA, Messenger ; genetics ; Transcription Factor RelB ; genetics ; metabolism ; Tumor Cells, Cultured

Objective: To study the specific killing effect of CD4 membrane protein targeted chimeric antigen receptor modified T (CAR-T) cell. Methods: The second generation CD4 targeted chimeric antigen receptor containing 4-1BB costimulation domain was insert into lentiviral vector through recombinant DNA technology. Lentivirus was prepared and packaged by 293T cells with four plasmids. Beads activated T cells were transduced with lentivirus and the transduction efficiency was checked with Protein L and flow cytometry. T cell subsets and IFN-γ concentrations were detected with probe-tagged antibody and cytometric bead assay. Results: ①The transduction efficiency of activated T cells with prepared lentivirus were 50.0%-70.0%. A subset of CD8⁺ T cell acquired dim expression of CD4 membrane protein after activation. CD4⁺T cell and CD8⁺CD4^dim T cell were gradually killed by CD4 targeted CAR-T post lentivirus transduction. ②The kill efficacy of CD4 targeted CAR-T cell and control T cell toward KARPAS 299 T cell at an E∶T ratio of 8∶1 for 24 h was (96.9±2.1)% and (11.2±3.1)%, CAR-T cell has a higher killing efficacy than control T cell (t=7.137, P=0.028). The IFN-γ concentrations in culture supernatant of CAR-T cell with K562-CD4 cell, CAR-T cell with K562 cell and CAR-T cell alone were (15 648±2 168), (1 978±354) and (1 785±268) pg/ml, CAR-T cell cocultured with K562-CD4 cell produced more IFN-γ than the other two controls (P<0.01). Conclusions CD4 targeted CAR-T has an immunophenotype of CD8⁺CD4^-T cell. CD4 targeted CAR-T cell has killing efficacy toward normal CD4⁺T cell and CD4⁺T lymphoma cell. CD4 targeted CAR-T cell also has a killing efficacy toward CD4^dim target cell.

Objective:To explore the application of venetoclax in transplantation of patients with refractory acute myeloid leukemia (AML).Methods:The diagnosis and treatment process of a patient with refractory AML who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) under venetoclax and hypomethylating agents bridging myeloablative preconditioning regimen after induction therapy failure in the First Affiliated Hospital of Soochow University in March 2020 were retrospectively analyzed.Results:The patient was a 28-year-old female who was diagnosed with refractory AML. The patient was initially given induction chemotherapy with IA (idarubicin+cytarabine) (3+7) regimen, but the disease did not relieve, then the induction chemotherapy with CLAG (cladribine+cytarabine+granulocyte colony stimulating factor) regimen was given, but the disease still did not relieve. After chemotherapy with venetoclax and hypomethylating agents bridging myeloablative preconditioning regimen, salvage haploid allo-HSCT was performed. Re-examination of bone marrow showed remission, and implantation was successful. The patient was followed up for 100 days and had sustained remission, and no transplantation complications occurred.Conclusion:For refractory AML patients who have failed primary induction therapy, the use of venetoclax and hypomethylating agents bridging myeloablative preconditioning regimen can be used as a preferred solution for salvage allo-HSCT.