Objective:To evaluate the effect of cryopreservation on clonogenic ability and apoptosis rate of mono-nuclear cells and CD34+cells in umbilical blood (UB), and to choose the index to present the freezing injury and optimize the cryopreservation of UB. Methods:hTe mono-nuclear cells (MNC) and CD34+cells were separated from UB and frozen.Atfer 30 days, they were thawed in warm water. Clonogenic capacity and clonogenic recovery before and atfer the cryopreservation was compared. We also used Annexin V-FITC-PI to investigate the apoptosis rate of the cells before and atfer the cryopreservation of these 2 types of cells. Results:hTe number of colony forming unit-granulocyte/monocyte (CFU-GMs) was not changed atfer freezing and thawing in both MNCs and CD34+cells, while the number of colony forming unit-granulocyte, erythrocyte, monocyte and megakaryocyte (CFU-GEMM) was obviously reduced after freezing in CD34+cells. The 2 types of cryopreserved cells had certain degree of apoptosis before the cryopreservation. MNC-type cryopreservation increased the cells apoptosis a little, while CD34+-type cryopreservation increased more. Conclusion:hTe cells have certain degree of apoptosis before the cryopreservation. hTe freezing and thawing procedure does affect the early stage progenitor cells-CFU-GEMM in the CD34+-type cryopreserved cells in UB. hTe damage may be induced by the cell apoptosis.

OBJECTIVE To investigate the effect and safety of teicoplanin to cure the neutropenic patients with infection.METHODS The 64 neutropenic patients with fever were treated with teicoplanin for empirical therapy,and then its effect and safety were evaluated.RESULTS Teicoplanin was effective in 53 patients,its effective rate was 88.3%,the adverse event rate was only 4.7%,and the adverse reactions were slight and reversible and it was well tolerant to patients.CONCLUSIONS Teicoplanin is one of the antibiotic agents with high performance,low toxicity and fast effect,which is an ideal agent for empirical therapy for neutropenic patients with fever in hematology units.

Objective To detect different animal erythrocyte hemolysins titer,and compare the application of these hemolysins in immunological experimental teaching,for selecting the better method of preparing high titer hemolysin for experimental teaching of medical immunology.Methods A total of 40 experiment rabbits were divided into 4 groups in this study,and immunized by sheep red blood cell(SRBC) and porcine red blood cell(PRBC) through different immunization procedures to prepare the hemolysin,detect and compare these 4 groups hemolysins titer by the complement hemolysis test.Results Rabbit Anti-SRBC in the group A was 1∶4 800,rabbit Anti-PRBC in the group B was 1∶1 200,rabbit Anti-SRBC in the group C was 1∶1 000,rabbit Anti-PRBC in the group D was 1∶200.Conclusion The hemolysin titer of the rabbit Anti-PRBC was lower than that of the rabbit Anti-SRBC by the same immunization procedures,and the immunization procedure by intradermal multi-point and auricular vein injection is the better method of preparing high titer hemolysin,so PRBC could replace SRBC as antigen,and immunize the rabbits for preparing hemolysin,which could be used in experimental teaching of medical immunology.

Objective To assess the safety and efficacy of endoscopic retrograde cholangiopancreatography (ERCP) for children with pancreaticobiliary diseases. Methods Data of children under 14 years old who have underwent ERCP in Nanjing Drum Tower Hospital between September 2007 and August 2016 were reviewed for completion, complications and therapeutic methods. Results A total of 41 children underwent 68 ERCP, including 6(8. 8%) diagnostic and 62(91. 2%) therapeutic procedures. All procedures were performed under deep sedation. Cannulation failed in only 1 child with anomalous junction of pancreaticobiliary duct. The procedure success rate was 98. 5%( 67/68 ) . There were 8 adverse events, including 7 mild post?ERCP pancreatitis and 1 fever. Incidence of adverse event was 11. 8%( 8/68) . There was no such severe adverse event as bleeding, perforation, death, or other anesthesia related adverse event. Thirty?two children ( 78. 0%) had follow?up, ranging from 2 month to 6 years. Children followed lived well with no long?term adverse event. Conclusion ERCP is an effective and safe procedure for the diagnosis and treatment of pancreaticobiliary diseases in children.

OBJECTIVE: To observe the effect of programmed cell death 5 (PDCD5) protein on the apoptosis of multiple myeloma KM3 cells induced by dexamethasone and to understand its mechanism. METHODS: The human recombinant PDCD5 (rhPDCD5) protein was added (alone of different concentrations or associated with dexamethasone) into KM3 cells. Cultured together for certain time, the cells were collected for the following experiments: (1)The effect of rhPDCD5 protein and dexamethasone on the apoptotic rate of KM3 cells was determined by flowcytometry (FCM) analysis after the cells were stained by Annexin V-FITC & PI (propidium iodide). (2)Caspase-3 activity of KM3 cells was evaluated by Western blot. (3)The expression of survivin protein in KM3 cells was detected by immunocytochemistry. RESULTS: The apoptotic rate of KM3 cells and the activity of caspase-3 increased significantly, and that treated with rhPDCD5 protein and dexamethasone was higher than that treated with rhPDCD5 protein only. The expression of survivin protein in the rhPDCD5 with dexemethas group was down-regulated, and with the concentration of rhPDCD5 and dexamethasone increasing, the changes was more obviously. CONCLUSION PDCD5 protein can induce the apoptosis of KM3 cells, and accelerate the apoptosis of KM3 cells induced by dexamethasone. PDCD5 protein may reduce the expression of survivin protein and increase activation of caspase-3 to play its role in promoting apoptosis.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Dexamethasone ; pharmacology ; Drug Synergism ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Multiple Myeloma ; pathology ; Neoplasm Proteins ; pharmacology ; Recombinant Proteins ; pharmacology ; Survivin

【Objective】 To study the technology of separating and purifying C1 esterase inhibitor (C1-INH) by using the waste washing liquid as raw materia during the preparation of human prothrombin complex (PCC) l. 【Methods】 C1-INH was isolated and purified by a two-step method of polyethylene glycol (PEG) 4000 precipitation and cation chromatography. The pH of raw materials and the concentration of PEG4000 were adjusted to investigate the optimal conditions of PEG4000 precipitation method. After PEG was precipitated and centrifuged, the supernatant is treated as the loading solution for cation exchange chromatography, using Fractogel EMD SE HiCap(M) gel and CM Sepharose FF gel for ion exchange chromatography. The most suitable gel and separation conditions were selected by comparing the C1-INH antigen yield, activity yield and specific activity. 【Results】 Under the condition of pH 6.1, when the mass fraction of PEG4000 was 14%, the recovery rate of C1 esterase inhibitor was close to 70%, and the removal rate of ceruloplasmin was more than 95% after stirring for 10 minutes. As fractogel EMD SE HiCap(M) gel was used for cation exchange chromatography, when the eluent salt concentration was 0.25 M sodium chloride, the activity yield of C1 esterase inhibitor was greater than 80%, and the specific activity was greater than 5 IU/mg. 【Conclusions】 Using the waste washing liquid as the raw material during the preparation of PCC, the C1 esterase inhibitor with high specific activity can be prepared through PEG precipitation and purification by Fractogel EMD SE HiCap(M) ion exchange chromatography.

Chronic myeloid leukemia (CML) is one of the most common hematological malignancies and characterized by the formation of Philadelphia (Ph) chromosome. Recently, tyrosine kinase inhibitors (TKI) treatment greatly improved the prognosis of CML. However, the options may be limited when a patient develops traditional TKI resistance or gene mutation. Herein, we reported a case. A 38-year-old male CML patient developed a BCR-ABL1 gene mutation of T315I after 2.5 years of TKI treatment, including imatinib and dasatinib. We adjusted the treatment with the combined application of dasatinib and axitinib. BCR-ABL1 gene copies dropped down and achieved an early molecular response at 2 months later. Subsequently, he received hematopoietic stem cell transplantation. Axitinib and dasatinib were applied for another half year after the allogeneic hematopoietic stem cell transplantation (allo-HSCT). Two years after the allo-HSCT, the BCR-ABL1 gene was still undetectable. It provided a successful example in treating CML patients carrying BCR-ABL1 T315I mutation via combination of axitinib with conditional TKI.
Adult ; Axitinib ; Dasatinib ; therapeutic use ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; Male ; Mutation ; Protein Kinase Inhibitors ; therapeutic use