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Objective To establish a stable lung cancer A549 cell line transfected by AMP-activated protein kinase (AMPK) expression vector,and to observe the effect of AMPK on proliferation as well as on the invasive ability of A549 cells.Methods Full-length of AMPK gene was amplified and its target gene was digested,then inserted into the GV358 plasmid.Co-tranfected 293T cells were subjected to the lentivirus equipment package.Subsequently,we collected the lentivirus supernatant to infect the A549 cells and establish a stably,overexpressed cell line A549.The mRNA and protein of AMPK were examined by real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting.The proliferation and invasion abilities of A549 cells were detected by methyl thiazolyl thiazolium (MTT) and Transwell assay.Results GV358-AMPK lentivirus vectors was successfully constructed by restrictive enzyme digestion and plasmid sequencing.There were significantly increased expressions of AMPK protein (5.87 times,P =0.002) and mRNA (16.12 times,P ＜ 0.001) after transfected with GV358-AMPK compared with the Vector group.Meanwhile,AMPK overexpression showed significantly lower proliferation (the forth day:0.53 ± 0.03 vs.0.64 ±0.05,P=0.021;the fifth day:0.58 ± 0.04 vs.0.80 ± 0.07,P =0.002) and weaken invasive ability [(1.6±0.5) ×l05 vs.(3.4±0.3) ×105,P=0.004] ofA549 cells.Conclusion The lentiviralAMPK expression vector and its A549 cell line is successfully constructed.AMPK overexpression inhibits the proliferation and invasive ability of A549 cells.
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Objective:To study the correlation between chronic urticaria and Helicobacter pylori infection,and to explore the significance of HP detection in the diagnosis and treatment of chronic urticaria. Methods: Totally 420 cases of chronic urticaria,who were treated in outpatient department from April 2014 to July 2015,and 450 cases of healthy physical people were selected randomly as healthy control group in the same period,then the serum HP unease antibody was detected by colloidal gold method,the positive rate of two groups patients with HP was analysed. Meanwhile 162 chronic urticaria patients with positive HP were divided into experimental group with 88 cases and control group with 74 cases. The patients in the control group were treated with conventional treatment,while the experimental group were treated with triple therapy based on the treatment of control group,and the clinical efficacy of different ther-apeutic methods was analysed in the chronic urticaria patients with positive HP. Results: The positive rate of HP in chronic urticaria group was 38. 6%, and the positive rate of HP in healthy control group was 14. 4%, the difference was statistically significant ( P<0. 05). The effective rate of clinical efficacy in the experimental group was significantly higher than the control group(P<0. 05). Con-clusion:There is close correlation between chronic urticaria and HP infection,HP detection has important clinical significance for the diagnosis and treatment of chronic urticaria.
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Objective:To investigate the expression of CD56 antigen in leukemia cells of the patients with acute myeloid leukemia (AML)and its relationship with the prognosis of AML, and to clarify the role of CD5 6 antigen expression in predicting the prognosis of the AML patients.Methods:171 AML (non-M3)patients aged from 14 to 60 years old,who received a IA Regimen as the first time inducing chemotherapy were chosen.Flow cytometric analysis was used to evaluate the CD56 expression in leukemia cells.COX proportional regression analysis was used to select the prognostic factors,and bivariable analysis was used to study the relationship between the positive rate of CD56 and overall survival (OS).The CD56+ group (n=52),including CD56≥50% expression group (n=39) and CD560.05),while the 2-year survival rate in CD56≥50% group was lower than that in CD560.05).The relapse rate and first year relapse rate of patients in CD56+ group (64.3% and 37.5%)were significantly higher than those in CD56- group (34.3% and 17.9% )(P0.05).The DFS in CD56+ group was shorter than that in CD56- group (P<0.05).The same DFS result was also found between CD56≥50%group and CD56<50% group (P<0.05).Conclusion:The expression of CD56 antigen in leukemia cells predicts a bad prognosis in the AML patients,and the higher expression of CD56 indicates the worse prognosis.
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Objective To evaluate the protective effectiveness of intranasal immunizations with recombinated-pneumococcal autolysin(Re-LytA), which protects mice against local and systemic Streptococ- cus pneumoniae(Sp) infection. Methods Testing group (group A): CpG as an adjuvant, the mice were intranasally immunized with purified Re-LytA, obtained by affinity chromatograph. The negative control group(group B) were intranasally immunized with sterile saline. And the positive control group (group C) were received 23-valent polysaccharide commercial vaccine through intramuscular injection. All the samples were collected 2 weeks post the last immunization. The levels of antibody was determined by ELISA. Then the mice were challenged intraperitoneally and intranasally with Sp, respectively. The infection and coloniza- tion was followed by monitoring colony-forming units of Sp in the blood, homogenized lung, and nasopharyn- geal lavage fluid 4 days post intranasal immunization. The mice were observed daily to note the livability of each group. Results The level of the LytA antibody (IgG, IgA, slgA) in group A were higher than that in group B and C (P < 0.05). Neither the LytA nor polysaccharide antibody could be detected in group B. Polysaccharide antibody could be detected in group C. After challenged intraperitoneally there was no signifi- cant difference in survival rates between group A and group C (P > 0.05), which was significant higher than that in group B (P <0.05). After challenged intranasally, compared with the group A, the geometric mean colony-forming units washed from the nasopharyngeal lavage fluid of the group B and group C were signifi- cantly higher (P <0.05). Conclusion lntranasal immunizations with Re-LytA can protect mice against lo- cal and systemic pneumococcal infection, and the protective immunity may be related to sIgA.
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Objective:To explore the Th1/Th2 type cytokines expression and its clinical significance in various clinical behaviors of HBV-infected individuals.Methods:The production of IFN-? and IL-4 by CD3+CD8+?CD3+CD8-cells in peripheral blood from normal donor and patients with hepatitis B virus infection were assessed by a four-color flow cytometry.Results:Compared with the healthy control,the percentage of Tc1 cells in AHB group was higher,whereas the percentage of Th1 and Tc1 cells in CHB group were lower,and lower than that of the AHB,but they increased significantly with the chronic hepatic inflammation activity.Conclusion:T-helper 1 type cytokines are predominant in peripheral blood cell of AHB and CHB,but the different percentage of Th1 type cytokines may be one reason of the final outcome of the HBV-infected individuals.