Objective To detect the change of interleukin-18 (IL-18) level in the serum of patients with acute leukemia (AL) in children, and explore the clinical significance of IL-18. Methods The level of IL-18 was measured by sandwich enzyme-linked immuno-sorbent assay (ELISA) in 45 patients with AL in children. Results The leverof IL-18 in pre-treatment AL group was 719.35±358.21pg/mL and significantly higher than that of normal-control group [(311.80±146.64)pg/mL P <0.01]. Mter treatment, the level of IL-18 was 401.14±180.78 pg/mL in post-treatment complete remission group, which was significantly lower than that of pre-treatment group(P <0.01). The level of IL-18 in non-remission group was higher than those of normal control and CR group (P<0.01);while pre-treatment group the difference was not significant (P >0.05). According to the clinical sub-group with risk factors in pre-treatment AL, the level of IL-18 in high risk(HR) and middle risk(MR) group was significantly higher than low risk(LR) group (P<0.05). The level of IL-18 in T-ALL group was significantly higher than that in B-ALL group (P<0.05). The levels of IL-18 in pre-treatment AL were markedly correlated to the count of blast cells in bone marrow (r=0.411, P=0.005). Conclusion The level of IL-18 in the patients of childhood AL was in a high expression, and related to the clinical treating effect and the count of blast cells in bone marrow, which would be taken as an index of treating effect. The level of IL-18 was closely related to the clinical risk factors in pre-treatment AL.

We established methods to isolate human amniotic fluid-derived progenitor cells (hAFPCs), and analyze the ability of hAFPCs to secrete human coagulation factor IX (hFIX) after gene modification. The hAFPCs were manually isolated by selection for attachment to gelatin coated culture dish. hFIX cDNA was transfected into hAPFCs by using a lentiviral vector. The hFIX protein concentration and activity produced from hAFPCs were determined by enzyme-linked immunosorbent assay (ELISA) and clotting assay. The isolated spindle-shaped cells showed fibroblastoid morphology after three culture passages. The doubling time in culture was 39.05 hours. Immunocytochemistry staining of the fibroblast-like cells from amniotic fluid detected expression of stem cell markers such as SSEA4 and TRA1-60. Quantitative PCR analysis demonstrated the expression of NANOG, OCT4 and SOX2 mRNAs. Transfected hAFPCs could produce and secrete hFIX into the culture medium. The observed concentration of secreted hFIX was 20.37% +/- 2.77% two days after passage, with clotting activity of 16.42% +/- 1.78%. The amount of hFIX:Ag reached a plateau of 50.35% +/- 5.42%, with clotting activity 45.34% +/- 4.67%. In conclusion, this study established method to isolate and culture amniotic fluid progenitor cells. Transfected hAFPCs can produce hFIX at stable levels in vitro, and clotting activity increases with higher hFIX concentration. Genetically engineered hAFPC are a potential method for prenatal treatment of hemophilia B.
Amniotic Fluid ; cytology ; Blood Coagulation ; Cell Culture Techniques ; Cell Separation ; methods ; DNA, Complementary ; Factor IX ; biosynthesis ; Genetic Engineering ; Genetic Vectors ; Humans ; Stem Cells ; cytology ; metabolism ; Transfection

A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.

BACKGROUND: Hemorrhagic cystitis remains a common complication of hematopoietlc stem cell transplantation.Granulocyte-macrophage colony stimulating factor (GM-CSF) affects proliferation and differentiation of hematopoietic stem/progenitor cells, adjusts functions of monocytes, granulocytes, lymphocytes and endothelial cells.OBJECTIVE: To investigate the protective effects of GM-CSF bladder irrigation in hemorrhagic cystitis after allogeneic hematopoietic stem call transplantation.DESIGN: Case analysis.PARTICIPANTS: A total of 15 hematopathy patients undergoing allogenic hematopoietic stem cell transplantation at the Zhongshan Hospital of Sun Yat-sen University from January 2004 to August 2006 (routine treatment group). A total of 16 hematopathy patients undergoing allogenic hematopoietic stem cell transplantation from September 2006 to December 2008 (GM-CSF group).METHODS: In the routine treatment group, patients received mesna, hydration, alkalization and forced diuresis in the prevention of hemorrhagic cystitis. In the GM-CSF group, GM-CSF was infused into the bladder in addition to mesna,hydration, alkalization and forced diuresis in the prevention of hemorrhagic cystitis 24 hours before cyclophosphamide treatment. Catheter was extracted 3 days following cyclophosphamide withdraw. Following washing with saline, the bladder was emptied. 10 mL of saline and 5 mL of lidocaine were added into 300 μg of GM-CSF. The mixture was infused into the bladder for 60-120 minutes.MAIN OUTCOME MEASURES: The following parameters were measured: occurrence of hemorrhagic cystitis and its correlation to graft versus host disease, as well as the occurrence of cytomegalovirus infection and urinary system infection.RESULTS: Compared with routine treatment group, the occurrence rate of hemorrhagic cystitis was significantly decreased in the GM-CSF group (x2=4.39, P < 0.05), mean duration of hemorrhagic cystitis and duration of hospitalization were significantly shortened (t=3.97, P < 0.05; t=3.13, P < 0.05), and the occurrence rate of over grade HI hemorrhagic cystitis was significantly reduced (x2=5.04, P < 0.05). Cystitis degree was associated with degree and duration of graft-versus-host disease (r = 0.76).Compared with the routine treatment group, cytomegalovirus infection rate was slightly decreased in the GM-CSF group (x2=0.28, P> 0.05), and occurrence rate of over grade Ⅲ hemorrhagic cystitis was higher in patients with cytomegalovirus infection.Compared with the routine treatment group, the occurrence rate of urinary system infection was slightly reduced in the GM-CSF group (x2=0.28, P > 0.05).CONCLUSION: GM-CSF bladder irrigation is well tolerated and often effective, and should be considered as a preparative regimen of hemorrhagic cystitis after allogeneic hematopoietic stem call transplantation.

The effect of oxymatrine on high-fructose-feeding induced insulin resistance and liver steatosis in rats was observed and the underlying mechanism in improving the hepatic lipid metabolism was explored.The results demonstrated that high fructose feeding decreased the glucose tolerance and increased hepatic lipid accumulation in rats,while oxymatrine could improve glucose tolerance and alleviate hepatic steatosis in rats.High fructose feeding stimulated the protein expressions of key lipid-synthesis enzymes,which were decreased by oxymatrine intervention.Both high fructose feeding and oxymatrine intervention had no significant effect on protein expressions of mitochondrial fatty acid oxidation enzymes.

A reporter system for ?C31 integrase was developed in NIH3T3 cells.The reporter plasmid coding green fluorescent protein(GFP) coupled with red fluorescent protein(RFP) was co-transfected with the plasmid coding ?C31 integrase, to show the activity of integrase in the cells.Fluorescence activated cell sorter(FACS) was used to measure the proportion of the cells containing red and green fluorescence.The increment of green cells was positively related to the increase in the transfection with plasmid coding ?C31 integrase.Approximately 90% of green cells were observed under a ratio of plasmid-?C31-integrase/reporter plasmid at 10∶1.This suggests that the ?C31 integrase reporter system provides a probe for the function of ?C31 integrase in cells.

BACKGROUND:Donor selection for high-risk acute leukemia is still controversial.OBJECTIVE:To compare the therapeutic efficacy of the unrelated donor peripheral blood and matched sibling allogeneic hematopoietic stem cell transplantation for high-risk acute leukemia.METHODS:Total 65 patients with high-risk acute leukemia treated during January 2008 to January 2016 were included,in which 30 patients chose the unrelated donor peripheral blood stem cell transplantation (UD), and other 35 chose the matched sibling allogeneic hematopoietic stem cell transplantation (MS) according to the wishes of patients and their own situation. After treatment, the chi-square test, Kaplan-Meier survival analysis method, and other methods were used to compare the implanted and hematopoietic reconstitution, the occurrence of graft-versus-host disease, relapse mortality and long-term survival between the two groups.RESULTS AND CONCLUSION:The implantation rate, platelet hematopoietic reconstitution time, the incidence of acute and chronic graft-versus-host disease, and its type exhibited no significant differences between the two groups (P > 0.05).The relapse rate, total death rate, and transplant-related mortality rates were 10.0%, 50.0%, and 40.0% in the UD group and 20.0%, 48.6%, and 25.7% in the MS groups, respectively, and the intergroup difference was insignificant (P > 0.05).The expected 2-year cumulative disease-free free survival and overall survival rates were (49.4±9.2)% and (52.6±9.2)% in the UD group and (53.9±8.5)% and (53.9±8.5)% in the MS group, respectively, and the intergroup difference was also insignificant (P > 0.05). Our experimental findings show that unrelated donor peripheral blood stem cell transplantation can be used as an effective alternative in the absence of sibling donors.

Objective:To analyze a novel epitope of Homo sapiens synapse associated protein and synthesize polyclonal antibody.Methods:FRG4 full-length sequence was obtained by PCR from human fetal liver library;by bioinformatics to detect the second structure of amino acids encoded by FRG4 and its epitope and motifs;by solid-phase peptide synthesis method to synthesize FRG4 peptides,then peptides were immunized to rabbits;by immunohistory to detect the expression of FRG4 in HepG_2 cells.Results:Select 13-peptides PKLVKEEVFWRNY by bioinformatics to synthesize rabbit anti-human FRG4 polyclonal antibody.Antibody purity was 82.79% and antibody dilution was 1∶16 000 detected by ELISA.The antibody had a good reaction and speciality in Western blot,it was mainly expressed in cytoplasm of HepG_2 cells.Conclusion:A novel Homo sapiens synapse associated protein(FRG4) antibody was synthesized successfully.

Objective To estimate the averaged excess relative risk(ERR) in Chinese population based on the radiogenic cancer risk of leukemia in Japanese atomic bomb survivor cohort,and to discuss proper method suitable for risk transfer between populations.Methods Based on BEIR Ⅶ radiogenic cancer model and population transfer model,and the 2009 Chinese leukemia baseline rates given in 2012 Chinese Cancer Registry Annual Report,comparison was made of population incidences in seveal countries to adjust the weighting factors.Results The ERR of three subtypes of leukemia as a whole was obtained,and the weighting factors for risk transfer model was assumed.The additive factor for male was 0.2,and the multiplicative factor was 0.8,while the additive factor for female was 0.15,and the multiplicative factor was 0.85.Conclusions For the risk transfer between populations,weighting factor was adjusted as a whole to obtain the ERR value for estimating the risk to Chinese population.The risk transfer method suitable for Chinese population was obtained by using the incidence rate available for Chinese population to directly transfer radiation-induced leukemia risk to Chinese from Japanese.

Objective To prepare a completely biological hybrid scaffold for small-diameter vascular tissue engineering using porcine fibrin and decellularized canine carotid artery.MethodsPorcine fibrin was sprayed coating on the external surface of decellularized canine carotid artery to construct completely biological hybrid scaffold for small-diameter vascular tissue engineering. The completely biological hybrid scaffold was evaluated with Hematoxylin and Eosin (H&E) staining, scanning electron microscopy and biomechanics test.ResultsHistology examination revealed that the porcine fibrin was sprayed coating uniformly on the external surface of decellularized canine carotid artery. Scanning electron microscopy examination confirmed that the external surface of completely biological hybrid scaffold was smooth and uniformly. Compared with fresh canine carotid artery and decellularized artery, the biological hybrid scaffold had similar burst and breaking strength. Furthermore, compared with decellularized artery, the biological hybrid scaffold had higher compliance.ConclusionThe porcine fibrin was sprayed coating uniformly on the external surface of decellularized canine carotid artery to prepare a completely biological hybrid scaffold for small-diameter vascular tissue engineering. The biological hybrid scaffold had appropriate biomechanical properties and had potential to serve as scaffolds for small-diameter vascular tissue engineering.