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 .
3.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.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.
6.Effect of femoral nerve catheter location on postoperative pain relief after total knee arthroplasty
Yan HU ; Feng ZHU ; Wei ZHANG ; Xiqiang HE ; Shizhong LI
Chinese Journal of General Practitioners 2011;10(10):749-751
We enrolled 60 patients with American Association of Anesthesiologists grade Ⅰ - Ⅱ undergoing unilateral total knee arthroplasty. All patients received combined epidural and spinal anesthesia,and a nerve stimulator was used to guide placement of a femoral nerve catheter. Patients were divided into three groups according to the catheter location on X-ray : psoas muscle group ( n = 18 ), iliacus muscle group (n = 19) and local group (n =23). Visual analog scale (VAS) pain scores were recorded at rest and with movement at 4, 24 and 48 h postoperatively and sensory blockade of the femoral, obturator and lateral femoral cutaneous nerves was recorded at 24 h.There were no significant differences in femoral nerve blockade among the three groups. Obturator nerve blockade was significantly better in the psoas muscle group than in the iliacus muscle and local groups, and was also better in the local group than in the iliacus muscle group. There was no significant difference in lateral femoral cutaneous nerve blockade between the psoas muscle and iliacus muscle groups, but there was better blockade in both these groups than in the local group. At 4 h postoperatively, VAS pain scores at rest were significantly lower in the psoas muscle group than in the iliacus muscle and local groups, but there were no significant differences in VAS pain scores with movement among the three groups. At 24 and 48 h postoperatively, VAS scores at rest and with movement were significantly lower in the psoas muscle group than in the iliacus muscle and local groups.
7.Assays of immunoreactive insulin and specific insulin in evaluating islet function and insulin sensitivity
Feng LI ; Lihong CHEN ; Juying TANG ; Li YAN ; Yang HE
Chinese Journal of Endocrinology and Metabolism 2008;24(2):184-185
Comparison of assays of immunoreactive insulin (IRI) and specific insulin in evaluating islet β cell function and insulin sensitivity suggested that there were no significant differences in individuals with different glucose tolerance impairment by two assays. The evaluation of islet β cell function using IRI and insulin sensitivity is still valid in clinical practice.
8.The Effect of the Simvastatin Administration on the Expression of Connective Tissue Growth Factor in Fibrotic Lungs of Rats
Yanhua FENG ; Li XIAO ; Hongguang WAN ; Yan DU ; Ying HE
Journal of China Medical University 2010;(9):719-723
Objective To explore the effect of the simvastatin administration on the expression of connective tissue growth factor (CTGF) in fihrotic lungs of rats. Methods The cats were treated with single intratracheal instillation of bleomycin (BLM) or instillation of the same volume of normal saline (NS) as a control. The administration of simvastatin(20 mg/kg)began once a day immediately or 7 days later after intratracheal BLM instillation respectively with the same volume of NS was given as a vehicle control. The rats were killed on day 7,14 and 28 respectively. Pathological alteration of lung tissue was observed hy HE staining and Masson staining. Hydroxyproline(HYP)content in lung tissue was used to determine the severity of pulmonary fibrosis. The expression of CTGF in lung tissue was exanfined by immunohistochem- istry staining and photodeusitometry. Results Histopathological changes of pulmonary fibrosis emerged gradually after the instillation of BLM. The expression level of CTGF was increased in lungs of rats after intratracheal instillation of BLM, compared with the control. The administration of simvastatin immediately or 7 days after intratracheal instillation of BLM, attenuated the histopathological changes of bleomycin- induced puhnonary fibrosis and prevented the increased expression of CTGF in lung tissue on day 28. Conclusion The adntinistration of simvastatin, immediately or 7 days after intratracheal BLM instillation, prevented the up-regulation of CTGF in fibrotic lungs of rats, which ntight be one of the mechanisms of the anti-fihrosis of simvastatin in lungs.
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.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.