The present experiment was to study the effect of stromal cell infusion on hematopoietic recovery after peripheral blood stem cell transplantation in mice. BALB/c mice pre conditioned with high dose chemotherapy/radiotherapy were infused with peripheral blood stem cells (PBSCT group) or a combination of peripheral blood stem cells and bone marrow stromal cells (experimental group). Four week survival rate, white blood cell (WBC) count, bone marrow nucleated cells (BMNC), granulocyte macrophage colony forming units (GM CFU), fibroblast colony forming units (F CFU) were determined. The results showed that survival rate, BMNC, GM CFU, and F CFU were significantly higher in experimental group than that in PBSCT group ( P

Objective This retrospective study was perform to assess risk factors of the type A aortic dissection after aortic valve replacement,in order to provide a basis for devising a strategy for future treatment.Methods From 2009 to 2012,23 patients were found type A aortic dissection and 20 patients were found aortic aneurysm after aortic valve replacement in Beijing Anzhen hospital.Analysis the clinical,pathology and surgery procedure characteristics of these patients,and assess the risk factors for type A aortic dissection after aortic valve replacement.Results After aortic valve replacement,There were significant different in age (P =0.012),bicuspid aortic valves (P =0.014),aortic valve regurgitation (P =0.009) and postoperative pathoanatomical characteristics(P =0.001) between type A aortic dissection group and ascending aortic aneurysm group.Multivariate logistic regression analysis indicated that age(P =0.032),aortic valve regurgitation(P =0.046) and pathoanatomical characteristics(P =0.002) were important risk factor for type A aortic dissection after aortic valve replacement,respectively.Conclusion Older age,aortic regurgitation combined and thinned or fragile aortic walls with systemic hypertension in patients with ascending aortic dilatation (≥45 mm diameter) at the time of AVR might be predisposing factors for postsurgical type A aortic dissection.These patients should be considered for concomitant replacement or repair of the ascending aorta unless the patient has a high operative risk.

Objective To analyze the practicality of current diagnostic criteria of invasive fungal infection (IFI) in patients with hematologic diseases/malignant tumors,so as to enhance the recognition of characteristics of pulmonary IFI after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods The clinical features of 51 cases with IFI after allo-HSCT were analyzed retrospectively.Results Pulmonary IFI accounted for 42.1％ (51/121) of the whole infectious pneumonia diagnosed among the patients admitted during the study.One (2.0％) case was proven diagnosis ; 24 (47.1％) were probable diagnosis and 26(51.0％) were possible diagnosis.The using of immuno-suppressors and corticosteroids,and the presence of graft-versus-host disease (GVHD) were the main host factors.The patients with two or more host factors simultaneously accounted for 66.7％ (34/51) of all pulmonary IFI patients.Totally 94.1％ (48/51) of the patients with pulmonary IFI presented nodules and/or patches as the main features in high resolution computed tomography (HRCT) scanning.The positive rates of fungal antigen detection were 58.6％ for G test and 33.3％ for GM test,which were relatively high.Twenty patients (39.2％) showed decrease of arterial partial pressure of oxygen and hypoxia in blood-gas analysis.Conclusions For the diagnosis of pulmonary IFI post allo-HSCT,the administration of immuno-suppressors and corticosteroids,and the presence of GVHD were the main host factors.Nodules and/or patches were the main features in HRCT image.Fungus antigen detection is the main tool to support clinical diagnosis.

Objective To analyze the efficacy and safety of HAA induction regimen consisted of homoharringtonine (HHT),cytarabine (Ara-C) and aclacinomycin (ACM) in naive and refractory relapsed acute myeloid leukemia.Methods Data from 66 acute myeloid leukemia (AML) cases hospitalized and treated with HAA induction regimen was analyzed retrospectively.Results 45 of the 66 cases suffered from naive AML,and 21 were refractory relapsed.HAA efficacy in naive AML was evaluated in 41 cases with 36 in complete remission (CR) and 1 in partial remission (PR).The efficiency of HAA induction regimen was 90.2 ％ (37/41)in naive AML group and 42.9 ％ (9/21) in refractory relapsed group,respectively.There were no differences (P ＞ 0.05) when considering patient' s gender,age,disease subtype and white blood cell count at onset.14 patients in CR with naive AML were followed-up for a median time of 9 months (2-17 months),and 5 cases relapsed (35.7 ％) in a range of 2-8 months.The median myelosuppression period was 14 days (3-23 days).Nausea and vomiting [20 ％ (13/66)] were the major side effects of HAA regimen,and the other side effects were abdominal pain and diarrhea [9 ％ (6/66).After chemotherapy,53 ％ (35/66) of the cases experienced infection/fever due to neutropenia.Other severe non-hematological side effects did not occur.Conclusion HAA regimen may be an ideal choice for the induction chemotherapy of naive and relapsed refractory AML.

AIM: To study the effect of mesenchymal stem cells (MSCs) infusion on hematopoietic recovery after peripheral blood stem cell transplantation in mice. METHODS: BALB/c mice conditioned by high dose chemotherapy/radiotherapy were infused with 10 6 peripheral blood mononuclear cells (PBMC) mobilized by granulocyte colony-stimulating factor (PBSCT group), 10 4 MSCs culture-expanded in vitro and 10 6 PBMC(experimental group 1), 10 6 MSCs and 10 6 PBMC(experimental gruop 2). Survival rate within 4 weeks, white blood cell count, bone marrow nucleated cells (BMNC), granulocyte-macrophage colony forming unit(GM-CFU) and fibroblast colony forming unit (F-CFU) were examined. RESULTS: Survival rate, BMNC, GM-CFU, F-CFU were significantly higher in experimental group 2 than that in PBSCT group ( P

BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P ＜ 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.

Objective To screen permethrin human single-chain variable region (scFv) antibody for aims of developing rapid detection kit. Methods Phage display technology was used in this study. Permethrin was solid phase coated on Nunc plate as antigen. Semi-synthetic single-chain variable region of human antibody library technology was applied, and single chain variable region was screened from phage antibody library after 3 rounds "adsorption - elution - amplification" of the selection process. 100 clones were random selected as resistance to permethrin clones , enzyme-linked immunosorbent assay (ELISA), crossreactivity and competitive inhibition experiments were used to validate permethrin binding activity with strong scFv clones from the selected phage antibody clones plasmid. The plasmid was digested with restriction enzyme Sfi Ⅰ / Not Ⅰ and subcloned into pCANTAB5E vector. After transformed into E. coli XL1BIue, the plasmid was identified by restriction enzyme analysis. Results After screening in 100 clones, 18 clones had high ELISA absorbance values ( A value) at 490nm wavelength ( A490nm), then bovine serum albumin (BSA) cross-reactions identified five weak cross-reaction. Combined with the triplicate ELISA and competitive inhibition experiment results, one positive clone was acquired at last. And this clone was subcloned into pCANTAB5E vector and transformed into competent cells XL1-Blue. Conclusion Plasmid fragment was consistent with the purpose, which provided the foundation for further study of its specific affinity.

A 46-year-old female patient was diagnosed as mixed phenotype acute leukemia with chief complaints of intermittent gingival swelling and bleeding for 1 week. The induction chemotherapy was not effective. During the second course chemotherapy, the patient had sudden convulsion and coma. She was transferred to the intensive care unit with worsened condition after transient improvement. Her final diagnosis was secondary adrenocortical insufficiency, adrenal crisis, intractable hyponatremia and cerebral edema.

Objective: To investigate the efficacy and prognostic factors of induction therapy in FLT3-ITD⁺ acute myeloid leukemia (AML) in the real world data. Methods: From January 2013 to December 2016, 114 de novo patients with FLT3-ITD⁺AML were enrolled in this study. Out of 114 cases, 75 were male, and 39 were female. The median age was 42 years old (ranged from 14 to 72 years old) . The chemotherapy regimens were used for induction therapy and all cases were followed up. The treatment response was evaluated by MICM and the comparison of the ratio were analyzed by chi-square test and the survival was estimated by Kaplan-Meier analysis and Cox proportional hazards model was used to identify independent prognostic factors. Results: There were 52 FLT3-ITD⁺AML patients with favorable prognosis genes (46 cases with NPM1, 5 cases with RUNX1-RUNX1T1, 1 case with CEBPA double mutation) and 62 patients with other types of FLT3-ITD⁺AML at diagnosis. All patients completed at least one cycle of induction therapy and the clinical curative effect was evaluated, complete remission (CR) rate was 50.0% (57/114) in one cycle and total CR rate was 72.5% (74/104) in two cycles. The CR rate of the FLT3-ITD⁺ AML patients with favorable prognosis genes was 67.3% (35/52) in one cycle and 83.3% (40/48) in two cycles; for the other types FLT3-ITD⁺AML patients, the CR rate was 35.5% (22/62) in one cycle and 64.8% (35/54) in two cycles. There was a significant difference in CR rate between the FLT3-ITD⁺AML patients with and without favorable prognosis genes (P<0.05) . This indicates that the FLT3-ITD⁺AML patients with favorable prognosis gene had relatively good therapeutic effect. Among other types of FLT3-ITD⁺AML patients who did not achieve remission from one cycle of chemotherapy, 9 patients were given sorafenib plus chemotherapy and 6 cases (66.7%) achieved CR; 23 patients were given conventional chemotherapy and 7 cases (30.4%) achieved CR. There was a significant difference between sorafenib plus chemotherapy and conventional chemotherapy groups (χ²=4.47, P<0.05) and this indicates that sorafenib plus chemotherapy can significantly improve the CR rate of FLT3-ITD⁺AML patients. Comparing overall survival (OS) and disease free survival (DFS) , there was no significant difference between sorafenib plus chemotherapy and conventional chemotherapy groups (P values were 0.641 and 0.517, respectively) . Conclusion The overall prognosis of FLT3-ITD⁺AML patients is poor, and the stratification therapeutic efficacy of FLT3-ITD⁺AML without favorable prognosis gene can be improved by sorafenib combined with chemotherapy.

Objective To explore the efficacy and safety of Sorafenib as monotherapy to FLT3 positive acute myeloid leukemia(AML).Methods From April 2014 to December 2015,fourteen AML patients with FLT3 positive,7 males and 7 females with a median age of 42(range:14-81) years old,were enrolled in this study.Of the 14 cases,4 were de novo cases,9 refractory cases and 1 relapsed case,including 78.6％ patients with severe complications and 57.1％ patients with KPS score less than 60 [the median KPS score was 45 (20-70)].The administration of Sorafenib was 400 mg twice daily and Sorafenib was continued if tolerated.The treatment response was evaluated by MICM and the data were analyzed by paired samples t test before and after Sorafenib treatment.Results The peripheral blood WBC count [4.2 (0.9-11.8) × 109/L vs 39.6 (2.3-209.5) × 109/L,P＜0.001],the percentage of peripheral blast cell [0.07 (0-0.54) vs 0.53 (0-0.94),P＜0.001] and the percentage of bone marrow blast cell [0.266 (0.020-0.880) vs 0.604 (0.180-0.900),P=0.003] were significantly decreased after Sorafenib monotherapy compared with before.The overall response rate was 57.1％ (8/14),including 5 cases (35.7％) with complete remission(CR).Of 4 de novo cases,2 achieved CR,1 with PR,1 with NR;3 of 10 refractory and relapsed patients achieved CR and 2 cases achieved PR,5 cases NR The median duration of achieving molecular remission (FLT3-ITD negative) after Sorafenib was 46 (33-72) days,and the median progression free survival (PFS) was 53 (28-175) days.Conclusion Sorafenib shows activity in FLT3-ITD mutation positive AML patients.Sorafenib monotherapy could be used as a treatment option for elderly patients or patients with severe complications,and refractory and relapsed patients with not suitable for intensive chemotherapy.