2.Effects of As2 O3 on human renal carcinoma cell line 786-O proliferation and PI3K-Akt transduction pathway
Feng ZHU ; Yan ZHANG ; Yan HE ; Huiqing ZHANG ; Yingjie ZHANG
Chongqing Medicine 2013;(26):3145-3148
Objective To evaluate the effect of arsenic trioxide (As2 O3 ) on the proliferation of human renal carcinoma cell line 786-O ,and to explore the changes of the PI3K-Akt signaling pathway .Methods Human renal cancer cells 786-O was cultured in 96-well plates ,and divided into the control group (n= 45 holes) and the experimental group (n= 45 holes) .After stimulation by 1 μM As2 O3 and saline ,the cells in 15 holes were collected at 0 ,12 ,and 24 h .BrdU assay was performed to quantify DNA synthesis to evaluate the cells proliferation ,the quantitative PCR was used to measure PI3K and Akt relative mRNA expression ,and Western blot was used to quantify the relative expression levels of of intracellular PI 3K and Akt .Results After 12 ,24 h of As2 O3 stimula-tion ,the amount of DNA synthesis in the observation group was gradually lower than that of the DNA synthesis at 0 h(P<0 .05) and significantly lower than that of the control group at 12 h and 24 h(P<0 .05) .At 0 ,12 ,24 h ,the relative expression level of in-tracellular PI3K and Akt mRNA and protein in the observation group had no significant difference (P>0 .05) ,and the relative ex-pression levels of PI3K and Akt mRNA and protein in the control group were increased as the proliferation was gradually increased . Conclusion As2 O3 inhibits human renal carcinoma cell line 786-O proliferation through inhibiting the PI3K-Akt transduction path-way ,and has potential clinical value for the treatment of kidney cancer .
4.Effects of cell-mediated immunity induced by intramuscular chitosan-pJME/ GM-CSF nano-DNA vaccine in BAlb/c mice.
Yong-Zhen ZHAI ; Yan ZHOU ; Li MA ; Guo-He FENG
Chinese Journal of Virology 2014;30(4):423-428
This study aimed to investigate the immune adjuvant effect and mechanism induced by chitosan nanoparticles carrying pJME/GM-CSF. In this study, plasmid DNA (pJME/GM-CSF) was encapsulated in chitosan to prepare chitosan-pJME/GM-CSF nanoparticles using a complex coacervation process. Immunohistochemistry was used to detect the type of infiltrating cells at the site of intramuscular injection. The phenotype and functional changes of splenic DCs were measured by flow cytometry after different immunogens were injected intramuscularly. The killing activity of CTLs was assessed using the lactate dehydrogenase (LDH) release assay. The preparation of chitosan-pJME/GM-CSF nanoparticles matched the expected theoretical results. Our results also found that, after pJME/GM-CSF injection, the incoming cells were a mixture of macrophages, neutrophils, and immature DCs. Meanwhile, pJME/GM-CSF increased the expression of MHC class II molecules on splenic DCs, and enhanced their Ag capture and presentation functions. Cell-mediated immunity was induced by the vaccine. Furthermore, chitosan-pJME/GM-CSF nanoparticles outperformed the administration of standard pJME/GM-CSF in terms of DC recruitment, antigen processing and presentation, and vaccine enhancement. These findings reveal that chitosan could be used as delivery vector for DNA vaccine intramuscular immunizations, and enhance pJME/GM-CSF-induced cellular immune responses.
Adjuvants, Immunologic
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administration & dosage
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Animals
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Chitosan
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administration & dosage
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immunology
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Dendritic Cells
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immunology
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virology
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Encephalitis Virus, Japanese
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genetics
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immunology
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Encephalitis, Japanese
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immunology
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prevention & control
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virology
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Female
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Granulocyte-Macrophage Colony-Stimulating Factor
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administration & dosage
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genetics
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immunology
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Humans
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Immunity, Cellular
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Japanese Encephalitis Vaccines
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administration & dosage
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genetics
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immunology
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Mice
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Mice, Inbred BALB C
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Nanoparticles
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administration & dosage
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Spleen
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immunology
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T-Lymphocytes, Cytotoxic
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immunology
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virology
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Vaccines, DNA
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administration & dosage
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genetics
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immunology
5.Effects of stellate ganglion block on erythrocyte immunity in patients with acute cerebral infarction
Chunjing HE ; Qian YU ; Yaping FENG ; Daiyi LIANG ; Yan RAN
Chinese Journal of Anesthesiology 2010;30(z1):46-49
Objective To investigate the effects of stellate ganglion block (SGB) on erythrocyte immunity in patients with acute cerebral infarction.Methods Twenty-four patients (13 male, 11 female) who developed acute cerebral infarction for less than 3 days were randomly divided into 2 groups (n=12each): Group A receiving traditional treatment and Group B receiving traditional treatment + SGB.The patients ranged in age from 51 to 64 yr and weighed 52-71 kg. All patients received intravenous 5% glucose 25 ml plus citicoline sodium 1.0 g and sodium ozagrel injectio 250 ml daily for 10 days in addition to dehydration and effective control of complications and intracranial pressure. Group B received SGB on one side alternatively with 1% licocaine 10 mi once a day for 10 days. Fasting venous blood samples were taken in the early mornings of the day before treatment (baseline, T1 ) and the 1st, 5th and 10th day of treatment (T2-4) for determination of the plasma MDA concentration and SOD activity, erythrocyte C3b receptor rosette rate (RBC-C3bRR) and RBC immune complex rosette rate (RBC-ICR) and Ne+-K+-ATPase activity in erythrocyte membrane.Results The plasma MDA concentration and RBC-ICR were significantly decreased during treatment es compared with the baselines at T1 in both groups (P<0.05 or 0.01), but were significantly lower in Group B than in Group A (P<0.05 or 0.01 ).The activities of plasma SOD and Na+ -K+ -ATPase in erythrocyte membrane and RBC-C3bRR were significantly increased during treatment as compared with the baselines at T1 and were significantly higher in Group B than in Group A.Conclusion SGB combined with traditional treatment can increase the activities of plasma SOD and Na+ -K+ -ATPase in erythrocyte membrane, inhibit production of oxygen free radicals and enhance RBC immune function in patients with acute cerebral infarction.
6.Establishment of co-culture model in vitro to induce bone marrow mesenchymal stem cells differentiate into lung epithelial cells
Yan WANG ; Zhijun YANG ; Xiyu HE ; Zhichun FENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2010;31(1):54-58,105
Objective To establish the co-culture model in vitro and induce bone marrow mesenchymal stem cells (MSCs) to differentiate into lung alveolar epithelial cells. Methods Each group had 6 samples, control group was MSCs alone; Group A was the MSCs cultured with the cells from normal lung; and Group B was the MSCs with the cells from injuried lung. Each group was cultured for 8 days and the two markers of lung alveolar epithelial cells including AQP5 and SP-C were tested by laser confocal microscopy and RT-PCR. Results Only AQP5 was detected in the control group and Group A, both AQP5 and SP-C were detected in Group B, the AQP5 mRNA expression in Group B was significantly increased compared with that in the control group(P<0.01). The AQP5 mRNA expression in Group B was also significantly increased compared with that in Group A (P<0.01). But there was no significant difference in AQP5 mRNA expression between Group A and control group. Conclusion We have successfully established the co-culture model in vitro to induce bone marrow mesenchymal stem cells to differentiate into lung epithelial cells.
7.High Throughput Screening Method and Application for L-glutamate Specific Aminotransferase
Linwei HE ; Zhangmin LIU ; Yan FENG ; Li CUI
China Biotechnology 2017;37(8):59-65
Objective:The aim is to establish L-glutamate specific aminotransferase-L-glutamate dehydrogenase coupling 96-well high throughput screening method,which is applied to molecular evolution of aminotransferase WecE from E.coli.Methods:An optical assay for aminotransferase catalytic activity based on aminotransferase-glutamate dehydrogenase coupling system is established by optimization of coupling enzyme loading,signal molecule NADH concentration and coupling time.Mutants library of WecE is obtained by sitedirected saturation mutagenesis.Positive mutants can be screened out through 96-well preliminary screening and flask second screening.Results:The target transamination reaction is coupled with L-glutamate dehydrogenase indicative reaction system which consists of 0.5 U/ml enzyme loading and 0.4 mmol/L NADH.A positive mutant Y321F whose catalytic activity increases 3.4 fold compared to that of wild type is screened out in Tyr 321 saturation mutagenesis library of WecE.Conclusion:An accurate high throughput screening method with weak background interference is established.It offers feasible solution for molecular evolution of L-glutamate specific aminotransferase.
8.Expression of caspase-3 survivin CyclinD1 and p27 in gastric carcinoma and their significance
Zhao-Xia YAN ; Quan-Hong WANG ; Feng-Ling HE ;
Cancer Research and Clinic 2006;0(10):-
Objective To study the expression of caspase-3,survivin,CyclinD1 and P27 in gastric carcinoma(CA2)and the relationship between the biomarkers and clinical pathological parameters. Methods Immunohistochemical Ready-to-use two-steps method was performed to research the expression of the four proteins in 72 cases of GC,and 54 cases of normal gastric mucosa.Results The expression level of caspase- 3 and p27 protein in GC was significantly lower than that in normal gastric mucosa(P
9.Development and evaluation of the heroin abstainers' cue-sensitization questionnaire.
Qian, HE ; Yan, FENG ; Ying, WANG ; Zengzhen, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(1):134-8
In order to develop the heroin abstainers' cue-sensitization questionnaire (HACSQ), and evaluate its reliability and validity, during March and April in 2006, the cross-sectional sample survey and cluster sampling were adopted among 939 volunteer participants who were heroin abstainers from 5 compulsive detoxification centers in China. The data were collected by the self-report questionnaires. The reliability analysis included Cronbach's alpha coefficient, split reliability and test-retest reliability, while construction validity was analyzed in the light of exploratory and confirmatory factor analysis. Based upon the exploratory factor analysis, 4 factors were extracted, which were social emotion cue (SEC), direct implement cue (DIC), personnel cue (PC) and indirect implement cue (IIC), and they together explained 64.52% variance. According to the data, test-retest reliability coefficient ranged from 0.57 to 0.63, and Cronbach's alpha coefficient ranged from 0.86 to 0.95 across the 4 factors and HACSQ. The adjusted goodness of fit index was 0.75. It was suggested that the HACSQ demonstrated good validities and reliabilities, and may be used as a reliable assessment tool for the cue-sensitization among the heroin abstainers.
China
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Cross-Sectional Studies
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Cues
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Factor Analysis, Statistical
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Heroin Dependence
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Questionnaires/*standards
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Reproducibility of Results
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Self-Assessment
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Young Adult
10.The Practice of Interactive Teaching in Medical Computer Basic Course
Zifeng LI ; Tianliang FENG ; Dasheng YAN ; Qing HE
Chinese Journal of Medical Education Research 2003;0(03):-
Guided by the principle of multimedia instruction, the author puts forward a new teaching mode, in which theoretical teaching and experimental teaching can be processed synchronously based on campus network, interactive teaching is dominative, students are the principal part, and integral optimization of theoretical teaching and experimental teaching can be realized.