Objective To investigate the effect of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) combined with recombinant bovine basic fibroblast growth factor (rb-bFGF) in the treatment of oral ulcer caused by chemotherapy.Methods 108 patients of oral ulcer caused by chemotherapy were randomly divided into two groups.54 cases in the control group were treated with cydiodine buccal tablets at first,then received the aerosol treatment which was prepared by mixing gentamicin,dexamethasone,2 ％ lidocaine and physiologic saline,three times per day.54 cases in the treatment group firstly received the gargle which was prepared by mixing rhGM-CSF,dexamethasone and physiologic saline,then were treated with rb-bFGF by spraying on the oral ulcer surface,three times per day.Results The effective rate of the treatment group was 96.30 ％ (52/54),which was significantly higher than that of the control group [64.81％ (35/54)],there was a significant difference between the two groups (x2 =17.08,P ＜ 0.05).Conclusion The effect of rhGM-CSF combined with rb-bFGF in the treatment of oral ulcer caused by chemotherapy is very significant.

Objective To construct the short hairpin RNA recombinant plasmids targeting mdr1 gene which expresses highly in ovarian cancer resistance strain SKOV3/TAXOL to silence endogenefic mdr1 gene expression and investigate the role of mdr1 gene in the development of resistant ovarian cancer. Methods The pGPU6/GFP/Neo-mdr1 were constructed by gene clone technology. The influence on proliferation and apoptosis were investigated by CCK-8 in SKOV3/TAXOL after transfected pGPU6/GFP/Neo-mdr1. Results The expression against mdr1 proteins were inhibited by pGPU6/GFP/Neo-mdr1. The cell proliferation were inhibited after transfected pGPU6/GFP/Neo-mdr1 by CCK-8. The apoptosis were observed in DAB experiments and the apoptosis rate increased. Conclusion mdr1 plays an important role in proliferation of resistant ovarian cancer and the short hairpin RNA of mdr1 can efficiently suppress mdr1 expression and enhance the apoptosis in SKOV3/ATAXOL.

Objective To construct the small hairpinRNA recombinant plasmids targeting mdr-1 gene which expresses highly in ovarian cancer resistance strain SKOV3/TAXOL to silence endogenetic mdr-1 gene expression and investigate the role of mdr-1 gene in the development of resistant ovarian cancer. Methods The pPGPU6/GFP/Neo-mdr-1 were constructed by gene clone technology. The influence on proliferation and apoptosis were investigated by CCK-8 in SKOV3/TAXOL after transfected. pPGPU6/GFP/Neo-mdr-1. Results The expression against mdr-1 proteins were inhibited by pPGPU6/GFP/Neo-mdr-1. The cell proliferation were inhibited after transfected pPGPU6/GFP/Neo-mdr-1 by CCK-8. The apoptosis were observed in DAB experiments and the apoptosis rate incised. Conclusion mdr-1 plays an important role in proliferation of resistant ovarian cancer and the short hairpinRNA of mdr-1 can efficiently suppress mdr-1expression and enhance the apoptosis in SKOV3/TAXOL, which provides a novel method for chemotherapy resistant tumors.

Flocculent gene FLO1 and its truncated form FLO1c with complete deletion of repeat unit C were expressed in a non-flocculent industrial strain Saccharomyces cerevisiae CE6 to generate recombinant flocculent strains 6-AF1 and 6-AF1c respectively. Both strains of 6-AF1 and 6-AF1c displayed strong flocculation and better cell growth than the control strain CE6-V carrying the empty vector under acetic acid stress. Moreover, the flocculent strains converted glucose to ethanol at much higher rates than the control strain CE6-V under acetic acid stress. In the presence of 0.6% (V/V) acetic acid, the average ethanol production rates of 6-AF1 and 6-AF1c were 1.56 and 1.62 times of that of strain CE6-V, while the ethanol production rates of 6-AF1 and 6-AF1c were 1.21 and 1.78 times of that of strain CE6-V under 1.0% acetic acid stress. Results in this study indicate that acetic acid tolerance and fermentation performance of industrial S. cerevisiae under acetic acid stress can be improved largely by flocculation endowed by expression of flocculent genes, especially FLO1c.
Acetic Acid ; chemistry ; Ethanol ; Fermentation ; Flocculation ; Glucose ; Industrial Microbiology ; Mannose-Binding Lectins ; genetics ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics

Objective To enrich breast cancer stem cells of breast cancer cell lines MCF-7 and MDA-MB-231 through culturing mammospheres,and to detect the expression differences of gene of breast cancer stem cells makers.Methods The MCF-7 and MDA-MB-231 cell lines were cultured by serum and serum-free medium.The proportion of CD44+/CD24-and CD133+ cancer stem cells was measured in cells derived from mammosphere cells or monolayer culture cells by flow cytometry,and the expression of CD44,CD24,CD133,ALDH3A1,ABCG2 and CXCR4 mRNA were detected by RT-PCR.Results Flow cytometry analysis indicated that the CD44+/CD24-low proportion of the MCF-7 mammosphere cells was higher than its adherent culture cells (P ＜ 0.05),and CD133+ proportion had no difference between them (P ＞ 0.05).However,the CD44+/CD24-/low proportion of the MDA-MB-231 mammosphere cells was lower than its adherent culture cells (P ＜ 0.05),while CD133+ proportion was significantly higher than its adherent cultured cells (P ＜ 0.05).RT-PCR analysis suggested that the expression of CD44 and ABCG2 increased obviously in MCF-7 microspheres (P＜ 0.05),and the expression of CD24,CD133,ALDH3A1 and CXCR4 had no significant difference between the mammosphere cells and adherent culture cells (P ＞ 0.05).The CD24,CD133,ABCG2,ALDH3A1 and CXCR4 increased obviously in MDA-MB-231 microspheres.On the contrary,the CD44 decreased (P ＜ 0.05).The expression of CD44 and CD24,CD133 and ALDH3A1 had significant differences in the microspheres of MCF-7 and MDA-MB-231 cells (P ＜ 0.05).Conclusion The related cancer stem cells gene expression is different in the microspheres of human breast cancer cell line,which indicates that the different subtypes of breast cancer may be derived from different origins.

Objective:To investigate the effect on proliferation and apoptosis of T-cell leukemia cells by silencing NRP-1 ( Jurkat cells).Methods:The lentivirus plasmid which expresses NRP1 gene specific shRNA was constructed in our preliminary ex-perimental.We transfected the lentivirus plasmid to human T-cell Lymphoma cells.The proliferation of Jurkat cells different groups and effect on cell proliferation after chemotherapy drug EPI-treated were found by CCK-8 kit.The proliferation level and apoptosis rate of the cells were detected by flow cytometry and Annexin-V-FITC/PI method.Results:The proliferation level of NRP-1 /shRNA interference group was decreased significantly in 48 h,72 h,96 h,which was compared with the control groups.The apoptosis rate of the NRP-1/shRNA interference group was increased compared with control groups.The chemotherapy drug sensitivity of epirubicin ( EPI ) test results showed that EPI concentration was 0.025,0.05,0.1,0.2,0.4 μg/ml,the NRP-1/shRNA interference group of cell growth inhibition rate was increased,the corresponding control group difference had statistical significance(P<0.05).We choose the drug con-centration of the EPI IC50 for next experiments.NRP-1/shRNA interference group cell apoptosis rate increased significantly after induction,compared with the control groups difference was statistically significant ( P<0.05 ).Compared with control group, the expression level of Bcl-2 protein was decreased and the expression level of bax protein was increased significantly after EPI induction.The percentage of cells at G0/G1 phase increased significantly,while those at S phase decreased significantly.Conclusion:Plasmid shRNA-NRP1 inhibited the expression of NRP1 in Jurkat cells and decreased the proliferation level of Jurkat cells and promote their apoptosis and enhance their drug sensitivity;the molecular mechanism may relate to down-regulation of Bcl-2 and up-regulation of Bax.and arrested the cell cycle at G0/G1 phase.

Objective To explore the therapeutic effects and adverse reactions of thalidomide in the treatment of peri-chemotherapy nausea and vomiting of cancer patients.Methods Total of 70 patients were randomly divided into two groups:the treatment group (38 cases) and the control group (32 cases).The treatment group was treated with thalidomide (oral administration at a dose of 100 mg per night,then dose can be added by 50 mg until the top dose of 200 mg per day).The original will be maintained if they cannot be tolerate of extensive dose.The treatment group was also injected 2mg tropisetron in 30 minutes before chemotherapy.The control group was only injected same dose tropisetron.All cases were examined antiemetic effects and evaluated adverse reactions.Results Nausea and vomiting control rates were 89.5 ％ (34/38) and 68.8 ％ (22/32) respectively in the treatment group and control group respectively with significant difference.The adverse reactions were similar between the two groups.Conclusion Thalidomide joint tropisetron can effectively control the peri-chemotherapy nausea and vomiting,and the adverse reactions can be acceptable.It could improve further indications of the drug.

Objective:To study the bioactivity of thymosin ?16 (T?16) in vitro and in vivo.Methods:Recombinant His-SUMO-T?16 was constructed and transformed into E.coli BL21(DE3) for induced expression.The product was treated by ultrasonication,ion-exchange chromatography and metal chelation chromatography respectively for purification.The fusion protein was cut by His-SUMO protease and then further purified by metal chelation chromatography and Superdex 30 gel chromatography.Results:Recombinant fusion protein His-SUMO-T?16 was soluble,whose specific activity was 5.3?105 U/mg.It could promote the proliferation of BALB/c 3T3 cells,rabbit corneal cells,and chicken embryo chorion vessels in vitro,and both the proliferation and migration of vascular endothelial cells in vitro were enhanced,and rabbit skin healing of alkali burns in vivo was accelerated.Conclusion:E.coli expressing vector of recombinant His-SUMO-T?16 fusion protein is constructed successfully,and recombinant protein T?16 has significant repairing effects.The study established a good foundation for further industrialization of T?16.

Objectives To investigate the effects of Apelin 13 on myocardial metabolism in diabetic rats.Methods A total of 40 male Wister rats were randomly divided into normal control group (NC,n=8) and experimental group (n =32).Diabetic rats model were induced by high-sugar and high-fat diet combined with low-dose intraperitoneal injection of streptozotocin (STZ).The wellestablished 28 diabetic model rats were randomly divided into diabetic model group (DM,n=14) and apelin-13 treated group (n=14).In the Apelin-13 group,diabetic rats were administered Apelin-13 [0.1 μmol/(kg · d)]by intraperitoneal injection for 10 weeks,while the control group and diabetic model group were given an equal volume of 0.9％ NaCl.At the end of the 10th week,all rats were sacrificed after fasting glucose measurement.Levels of serum free fatty acids (FFA) and myocardial FFA were measured by ELISA.Expression of myocardial glucose transporter member 4 (GLUT4) were detected by immunohistochemistry.The mRNA expressions of myocardial PPARα,CD36 and CPT-1 were detected by real-time fluorescence quantitative PCR.Results Fasting blood glucose,serum FFA and myocardial FFA were significantly higher in DM group than in NC group (all P＜ 0.05).The level of plasma glucose and myocardial FFA were significant lower(P＞0.05) in Apelin-13 treated group than in DM group;but serum FFA was not significantly lower(P＜0.05).The mRNA expressions of PPARα,CD36,CPT-1 in cardiac myocyte were higher in DM group and Apelin-13 treated group than in control(P＜0.05),and lower in Apelin-13 treated group than in DM group(P＜ 0.05).The expression of myocardial GLUT4 was significantly lower in DM group(1.138±0.316)and in Apelin-13-treated group (4.631 ± 1.832) than in NC group(9.132 ± 2.156),(F=65.507,P＜ 0.05),and higher in Apelin-13-treated group than in DM group(P＜0.05).Conclusions Apelin-13 increases myocardial expression of GLUT4,improves utilization of FFA,and it can effectively reduce the expression of PPARα,CD36 and CPT-1.Therefore,it may play a vital role in the improvement of myocardial metabolism in diabetic rats.

Objective To study the changes and mechanism of the function of islet βcells and insulin signal transduction molecules in rats after long-term period lipid infusion.Methods Thirty SpragueDawley (SD) rats were randomly divided into free fatty acid (FFA) and normal saline (NS) groups.Catheters were implanted under pentobarbital anesthesia in the right atrium via the jugular vein and the left carotid artery.A technique for a 72-h infusion in unrestrained rats was used for triglyceride and heparin or saline infusion.The infusion period was started on day 2 after surgery.After 72-h infusion,fasting serum insulin (Ins) and FFA in the blood were determined.The glucose infusion rat (GIR) was measured by hyperinsulinemia euglycemic clamp to evaluate the peripheral insulin resistance.The intravenous glucose tolerance test (ivgtt) and islet cell perifusion was conducted to evaluate the function of islet β-cell.The rats in two groups were sacrificed,and the pancreatic islets were isolated and collected.The levels of malondialdehyde (MDA) and reduced glutathione hormone (GSH) were detected in pancreatic tissues.The expressions of insulin receptor substrate-1 (IRS-1),insulin receptor substrate-2 (IRS-2),and glucose transporter-2 (Glut2)gene in islets were detected by real-time polymerase chain reaction (PCR).Results (1)The serum FFA concentration in the FFA group was higher than in NS group [(1.56 ± 0.21) mmol/L vs (0.65 ± 0.12)mmol/L,P ＜0.01].(2)The GIR was decreased significantly in FFA group compared with NS group(P ＜0.01).(3)The glucose that stimulated insulin secretion was decreased in the FFA group.(4)The levels of MDA were significantly higher in FFA group [(1.62 ± O.18) mmol/mg prot vs (0.76 ± 0.15) mmol/mg prot,P ＜0.01].The levels of GSH were lower in FFA group [(22.54 ±2.66) mg/g prot vs (36.58 ± 3.02) mg/g prot,P ＜ 0.01].(5) The gene cxprcssion of IRS-1 in islets was significantly decreased by [(36.8±1.8)％,P ＜0.01],and the expression of IRS-2 and Glut-2 was decreased by [(29.6±1.2) ％] and [(58.7 ± 2.1) ％] in FFA group,respectively(all P ＜0.01).Conclusions Lipid infusion in long time decreased the secretion of insulin and impaired the expression of insulin signal transduction molecules in islet βcells.