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.Comparative study of bovine and porcine derived materials in hydrolysate samples by real-time fluorescence quantitative PCR and general PCR
Yan YU ; Suting HE ; Ziqiang WANG ; Feng DENG
Chinese Journal of Biochemical Pharmaceutics 2015;37(5):18-20
Objective To compare real-time fluorescence quantitative PCR with general PCR in detecting bovine and porcine derived materials in hydrolysate samples.Methods DNA were extracted from hydrolysate samples which prepared by different steps by real-time fluorescence quantitative PCR and general PCR.Results DNA of bovine and porcine could be detected by real-time fluorescence quantitative PCR and general PCR in samples prepared in the processes before enzymolysis solution, but not detected in samples from supermatant to the fourth ultrafiltrate.Conclusion Both real-time fluorescence quantitative PCR and general PCR can be applied to detect the fragments in hydrolysate samples.And real-time fluorescence quantitative PCR has the advantage such as rapid,convenient, non-environment-polluted, good repeatability, which improves the quality and efficiency.
6.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.
7.Accuracy of point-of-care testing for blood glucose monitoring in critically ill patients:evaluation of glucase oxidase and glucose dehydrogenase methods
Tao FENG ; Lijuan HE ; Xiangyuan CAO ; Hong YAN
Chinese Journal of Anesthesiology 2013;(1):58-61
Objective To evaluate the accuracy of point-of-care testing (POCT) for blood glucose monitoring in critically ill patients.Methods Two hundred and forty critically ill patients,of both sexes,aged 20-88 yr,with Acute Physiology and Chronic Health Evaluation Ⅱ score of 1-45,were enrolled.The venous,arterial and capillary blood samples were collected to determine the real-time blood glucose level using glucose oxidase (GOD) and glucose dehydrogenase (GDH) methods.The blood glucose level measured by central laboratory hexokinase method simultaneously was served as standard level.Error Grid analysis (EGA) and Bland-Altman analysis were used to determine accuracy and consistency,respectively.The accuracy of real-time blood glucose levels within the consistent limits was evaluated.Results 1.The results of EGA showed that 98.7 %,98.3 %,98.3 %(GDH method) and 96.2%,96.6%,96.7% (GOD method) of the difference between venous,arterial and capillary blood glucose levels measured and the standard level were located in the A and B zones,respectively,and 1.2%,1.7%,1.7% (GDH method) and 2.9%,3.3%,3.3% (GOD method) in the D zone.0.8% (GOD method) of the difference between venous blood glucose levels and the standard level were located in the C zone.2.Bland-Altman analysis showed that the difference between the standard level and glucose level measured in blood samples from the vein,artery and capillary.was-0.1,-0.3,-0.2 mmol/L (GDH method) and-0.9,-1.0,-0.9 mmol/L (GOD method),respectively,and the incidence beyond the upper and lower limits of consistency zone was 4.5 %,6.7 %,6.6 % (GDH method) and 4.6 %,5.0 %,7.1% (GOD method),respectively.The accuracy of venous,arterial and capillary blood glucose levels within the consistent limits was 94.3 %,92.1%,93.7% (GOD method) and 96.6%,95.1%,95.5% (GDH method),respetively.Conclusion The accuracy of POCT for blood glucose monitored by GOD and GDH methods is good in critically ill patients,but it is possible to overestimate the patient's real glucose level.
8.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.
9.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.
10.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.