1.Antagonism of geniposide on Toll-like receptor 7/nuclear factor-κB signaling pathways in cells with influenza A virus infection
Chinese Journal of Microbiology and Immunology 2010;30(8):749-754
Objective To observe how geniposide as an anti-inflammatory agent through inhibition Toll-like receptor 7/nuclear factor-κB signaling pathways activation, as well as TNF-α and IL-6 release infectioned by influenza virus. Methods Epithelial cells was exposed to human influenza viruses A/Gui/81/23(H3N2) infection for 2 h before treatment with geniposide for 24 h. NF-κB responsive element luciferase reportor gene was transfected and dual luciferase cis-reporting systems was used to assay the transcriptional activity of NF-κB under the stimulated circumstance of influenza virus infection. The phosphory level and nuclear transposition of NF-κB was observed by fluorescence inverted microscope. RT-PCR was used to detect the gene transcription level of TLR7, TNF-α and IL-6. Results The relative luciferase reporter assay of NF-κB was apparently improved by influenza virus infection. But geniposide significantly repressed the relative value of luciferase. The phosphorylation level and rate for nuclear transposition of NF-κB was apparently improved by influenza A virus infection observed by fluorescence inverted microscope. But geniposide significantly repressed the phosphorylation level and rate for nuclear transposition. RT-PCR showed upregulation of TLR7 and pre-inflammatory markers TNF-α and IL-6 in A549 cells infected by influenza virus, geniposide had a significant effect on the expression of TLR7 and inflammatory markers TNF-α and IL-6 after treated with influenza virus. Conclusion Geniposide as an antiinflammatory agent antagonized influenza A virus infection through inhibiting Toll-like receptor 7/nuclear factor-κB signaling pathways activation, as well as on the downregulation of the downstream inflammatory markers target gene expression TNF-α and IL-6.
2.Effect of stellate ganglion block on cerebral vasospasm in patients undergoing intracranial aneurysm surgery
Fangxiang ZHANG ; Shuixiang YIN ; Chunjing HE
Chinese Journal of Anesthesiology 2011;31(6):729-731
Objective To evaluate the effect of stellate ganglion block (SGB) on cerebral vasospasm in patients undergoing intracranial aneurysm surgery. Methods Forty ASA Ⅱ or Ⅲ patients aged 14-64 yr weighing 40-81 kg undergoing intracranial aneurysm clipping were randomly divided into 2 groups ( n = 20 each): group control (group C) and group SGB. Left SGB was performed with 0.25% ropivacaine 10 ml immediately after intubation. Successful block was verified by development of Homer syndrome within 15 min after block. Anesthesia was induced with midazolam, propofol, fentanyl and vecuronium and maintained with isoflurane inhalation and intermittent iv boluses of fcntanyl and vecuronium. The patients were intubated and mechanically ventilated. PETCO2 was maintained at 30-35 mm Hg. BIS was maintained at 50-60. Right internal jugular vein was cannulated and the catheter was threaded cranially until resistance was met for blood sampling. Blood samples were collected before skin incision (T1), before clipping of aneurysm (T2), at 30 min after clipping (T3 ), and at the end of surgery (T4) for determination of plasma concentrations of endothelin (ET), calcium gene-related peptide (CGRP) and S100B protein. Transcranial Doppler was used to measure the flow rate of blood in bilateral middle cerebral artery and extracranial carotid artery at 1 and 3 days after surgery. All patients were observed for incidence of brain ischemia during 1-7 days after surgery. Results Plasma ET and S100B protein concentrations were significantly decreased, while plasma CGRP concentration was significantly increased after clipping of aneurysm at T3 and T4 in group SGB as compared with group C. The incidence of cerebral vasospasm and brain ischemia was significantly lower in group SGB than in group C. Conclusion SGB performed before operation can significantly reduce the incidence of cerebral vasospasm after clipping of intracranial aneurysm by inhibiting the release of ET and promoting the release of CGRP.
3.Effects of Influenza virus H3N2 on TLR7 signal pathyway and intervention of Dureping Injection in vitro
Xuyan NIU ; Chunjing ZHANG ; Ligang GU
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(02):-
0.05). Compared with control cells, the relative luciferase activity of NF-?B in virus-infected cells was apparently up-regulated (P
4.The Effect of CIK Cell on Apoptosis of Lung Cancer Cell A549
Yan SHI ; Chunjing ZHANG ; Xiucai LIU
Journal of Medical Research 2006;0(05):-
Objective To investigate the inhibitory effects of CIK cell on apoptosis of lung cancer cell A549 in vivo. Methods CIK cells were induced by culturing PBMC with regular method and cell apoptosis was detected.Results Electron microscopic observations showed that CIK cells could induce the lung cancer cells A549 to apoptosis. Flow cytometry(FCM)demonstrated that apoptosis cells of lung cancer cells A549 were increased in CIK group as compared with the control group. Conclution CIK cells can induce the apoptosis of lung cancer cells A549.
5.Significance produced by examining NSCLC vein formation through enhanced SCT scanning in clinical treatment
Qiang ZHANG ; Chunjing LI ; Zhaoxin LIU ; Zhuang TIAN
Cancer Research and Clinic 2009;21(2):114-116
Objective To evaluate the examination of non-small-cell lung (NSCLC) vein formation and the dynamic characteristics of blood stream through enforced SCT scanning for the research on and clinical diagnosis of tumor.Methods Double period enhanced SCT scanning was done to 152 NSCLC cases identified pathologically.Makes the color coding the tumour blood stream irrigation chart,the analysis blood stream irrigation characteristic.Compare the enforced morphologic manifestation of cancer focus with histology and analyze their pertinence.Results The enforced CT peak value (PV) of the low differentiation is bigger than that of the medium differentiation,which is bigger than the high differentiation.63 cases are 45-70 HU,78 cases are 20-45 HU and 11 cases are 10-20 HU.67 case are of abnormal arteriola; 23 cases are of abnormal hemal sinus development; 35 cases are pistil-like.Conclusion Accurately examining and quantifying cancer focus vein formation according to SCT double period enforced scanning is of high guiding value in the enactment of the plan for treating NSCLC and the comprehensive treatment of tumors.
6.Effect of ginsenoside Rg1 on the transcriptional activation of NF-κB induced by H_2O_2 in 293T cell
Chunjing ZHANG ; Ligang GU ; Xuyan NIU ; Fu WANG ; Guiying PENG
Chinese Journal of Immunology 2009;25(11):991-995
Objective:To observe the influence of ginsenoside Rg1 on transcriptional activation of NF-κB induced by hydrogen peroxide (H_2O_2) in 293T cell,and probe into the antioxidant mechanism of ginsenoside Rg1.Methods:In the experiment,cells was exposed to H_2O_2 after pretreatment with Rg1,cell proliferation and cytotoxicity studies were detected by MTT and Trypan blue.The quantities of generation of intracellular reactive oxygen species (iROS) was analyzed by flow cytometric analysis measured with fluorescent probe 2,7-dichlorofluorescin diacetate (DCFH-DA).NF-κB-responsive element-luciferase reporter gene was transfected and dual-luciferase cis-reporting systems were used to assay the transcriptional activity of NF-κB under the stimulated circumstance of oxidative stress induced by H_2O_2.Results:The results of MTT showed that ginsenoside Rg1 apparently protected the proliferation of 293T cell,which were repressed by H_2O_2 (P<0.05).The results by trypan blue showed that H_2O_2 stimulated substantial cytotoxicity.This effect was markedly attenuated by treatment with ginsenoside Rg1.Oxidant production,measured as the fluorescence of dichlorofluorescein,was significant increased by 40%-50% through H_2O_2 stimulation.The decrease in iROS generation was significant blocked by 35%-40% through Rg1 and antioxidant.The relative luciferase reporter assay of NF-κB was apparently improved by H_2O_2-induced(P<0.05),but Ginsenoside Rg1 significantly repressed the relative value of luciferase (P<0.05).Conclusion:Ginsenoside Rg1 has the obvious protective function from the damage of oxidative stress damage,whose possible mechanism is to eliminate excessive free radicals of the cells effectively,to reduce transcriptional activation of nuclear factor kappa B(NF-κB),and subsequently to suppress the NF-κB circuit activation.
7.The effect of P-glycoprotein on 18F-FDG uptake in tumor-bearing nude mice
Chunjing YU ; Bin ZHANG ; Shengming DENG ; Weixing WAN ; Yiwei WU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;35(3):208-211
Objective To evaluate the relationship between 18F-FDG uptake and P-gp expression in Bcap37 or Bcap37/MDR1 tumor-bearing BALB/c nude mice.Methods Bcap37 or Bcap37/MDR1 cells were injected into BALB/c nude mice (1× 107cells/ml,0.2 ml/mouse) to construct mice models.Bcap37 (n=5) or Bcap37/MDR1 (n=5) tumor-bearing mice fasted for 6 h before imaging.After anesthesia,the mice were injected with 7.4 MBq of 18F-FDG via tail vein.The dynamic microPET scans were carried out for 90 min.On the microPET images,the ROI was drawn and the TAC was obtained.The next day,those 10 mice underwent dynamic microPET scans after injected with elacridar (GF120918) and 18F-FDG.Another 10 mice,5 with Bcap37 tumors and 5 with Bcap37/MDR1 tumors,were used.After 7.4 MBq 18F-FDG with or without 2.0 mg/kg GF120918 was administered via tail vein,microPET images were acquired at 60 min.ROI was drawn over the tumors and SUV was obtained.Two-sample t test was used to analyze the data.Results GF120918 did not significantly alter the 18F-FDG accumulation curve in Bcap37 tumors,but significantly enhanced the 18F-FDG accumulation in Bcap37/MDR1 tumors.GF120918 did not influence 18F-FDG uptake (SUV) in Bcap37 tumors (1.052±0.028,1.028±0.045,t =1.792,P>0.05),but significantly increased the SUV in Bcap37/MDR1 tumors (1.015±0.043,0.712±0.031,t=3.365,P<0.05);The SUV of 18 F-FDG in Bcap37 tumors was significantly higher than that in Bcap37/MDR1 tumors without injection of GF120918 (t =3.952,P<0.05).The SUV was not significantly different when GF120118 was injected (t=1.835,P>0.05).Conclusions 18F-FDG is a substrate of P-gp.18F-FDG imaging combined with GF120918 injection may be an effective noninvasive method for the detection of tumor's MDR.
8.The effect of P-glycoprotein on 18F-FDG uptake in vitro
Chunjing YU ; Bin ZHANG ; Shengming DENG ; Weixing WAN ; Yiwei WU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2013;(3):213-216
Objective To evaluate the effect of P-gp inhibitors of verapamil (VER) and GF120918 on 18F-FDG uptake in Bcap37 and Bcap37/multidrug resistancce (MDR)1 cell lines in vitro,and to explore the relationship between 18F-FDG uptake and P-gp expression at cellular level.Methods Bcap37 and Bcap37/MDR1 cells were seeded into 6-well plates at a density of 1 × 106 per well.Three days later,37 kBq/ml 18F-FDG,or 37 kBq/ml 18F-FDG + 100 μmoL/L VER,or 37 kBq/ml 18F-FDG + 50 μmol/L GF120918 were added into each well.Mter incubated for 10,30,60 and 120 min at 37 ℃ and in 5% CO2,the medium was removed and the cells were washed three times with 1 ml ice-cold PBS immediately.The radioactivity of 18 F-FDG was measured using a gamma counter.The uptake of 18F-FDG was expressed as the ratio of 18F-FDG radioactivity in Bcap37 or Bcap37/MDR1 cells and the overall radioactivity added to the cells in each well.The t test was used for statistical analysis.Results 18F-FDG uptake was higher in Bcap37/MDR1 cells than that in Bcap37 cells after incubated for 10 min.The uptake rate was (1.88 ±0.19) % in Bcap37/MDR1 cells and (1.37 ± 0.18) % in Bcap37 cells (t =7.832,P < 0.05).On the contrary,18 F-FDG uptake was significantly higher in Bcap37 cells than that in Bcap37/MDR1 cells after incubated for 60 and 120 min.The uptake rates were (2.29 ±0.23)% and (2.34 ±0.15)% in Bcap37 cells,(1.47 ±0.14)% and (1.53 ±0.22)% in Bcap37/MDR1 cells (t =8.437,8.283,both P < 0.05).18 F-FDG uptake was significantly higher with VER or GF120918 in Bcap37/MDR1 cells than that without VER or GF120918 after the incubation of 60 and 120 min (t =9.032,9.243 and 8.765,8.803,all P < 0.05).The uptake rates with VER or GF120918 were (2.45 ±0.21)% and (2.46 ±0.25)%,(2.50 ±0.24)% and (2.48 ±0.27)%.There was no significant difference of 18F-FDG uptake in Bcap37 cells with or without VER or GF120918.Conclusions 18F-FDG is a substrate of P-gp at cellular level.P-gp may act as an efflux pump to reduce 18F-FDG uptake in Bcap37/MDR1 cells.The uptake of 18F-FDG can be used to evaluate the function of P-gp in tumor cells.
9.Comparison of occurrence of postoperative cognitive dysfunction in elderly patients using different anesthetic methods
Fangxiang ZHANG ; Junping NING ; Bing QIU ; Shiping WANG ; Chunjing HE
Chinese Journal of Anesthesiology 2013;(2):188-190
Objective To compare the occurrence of postoperative cognitive dysfunction (POCD) in elderly patients using differentanesthetic methods.Methods Ninety-three ASA Ⅱ or Ⅲ patients,aged ≥ 65 yr,weighing 45-67 kg,scheduled for artificial femoral head replacement,were randomly divided into 2 groups:general anesthesia (group G,n =47) and combined spinal-epidural anesthesia group (group S-E,n =46).In group G,anesthesia was induced with iv injection of midazolam 0.1 mg/kg,propofol 2 mg/kg,fentanyl 3-5μg/kg,and vecuronium 0.1 mg/kg,and maintained with continuous infusion of propofol 2-3 mg· kg-1 · h-1,intermittent iv boluses of fentanyl 1 μg/kg and vecuronium 0.04 mg/kg and inhalation of 1.5%-2.0% isoflurane.In group S-E,hyperbaric 0.5 % ropivacaine 2 ml was injected into the subarachnoid space over 20 s,the patients were kept in the original position for 15 min,the level of anesthesia was simultaneously adjusted to below T8 on the operated side,and 0.5 % ropivacaine 3-5 ml was injected into the epidural space when needed during operation.Cognitive function was assessed by mini-mental state examination at 24 h before anesthesia and 24 and 72 h after operation.Venous blood samples were collected for determination of plasma amyloid-beta levels by ELISA.Results Compared with group G,the incidence of POCD at 24 h after operation and level of plasma amyloid-beta were significantly decreased in group S-E (P < 0.05).Conclusion Elderly patients are more likely to develop POCD under general anesthesia than under combined spinal-epidural anesthesia.
10.Overexpression of N-terminal amino acids of p55γregulatory subunit of PI3K inhibits cell adhesion of human gastric carcinoma MGC803 cells
Hongyan GUO ; Xiaodan QI ; Chunjing ZHANG ; Qi WU ; Xiaojie SUN
Chinese Journal of Pathophysiology 2015;(9):1563-1567
AIM:To investigate the effect of the N-terminal 24-amino acid (N24) overexpression in p55γre-gulatory subunit of phosphatidylinositiol 3-kinase ( PI3K) on the cell adhesion of human gastric carcinoma cell MGC 803. METHODS:MGC803 cells, which stably expressed GFP-N24 fusion protein and GFP alone , were generated by transfec-tion with pEGFPN-24 plasmid and control plasmid pEGFP-C1, respectively.The morphological change of the cells was ob-served under inverted microscope .The expression of GFP-N24 fusion protein was detected by Western blot .The adhesion of the cells was determined by cell adhesion assay .The effects of GFP-N24 fusion protein on the expression of E-cadherin andβ-catenin were analyzed by Western blot .The expression and secretion of matrix metalloproteinase 9 (MMP9) and u-rokinase-type plasminogen activator ( uPA ) in the MGC803 cells were detected by gelatin zymography .RESULTS:MGC803/GFP-N24 cell line steadily expressed GFP-N24 fusion protein and MGC803/GFP cell line steadily expressing GFP were successfully established , but the expression of fusion protein GFP-N24 was lower than that of the control protein GFP.The morphological changes of the transfected cells from paving stone to fibroblast cell form after gene transfection , and the cytoplasm secretory granules were increased significantly .The cell adhesion to fibronectin and collagen decreased after GFP-N24 transfection .GFP-N24 fusion protein increased the expression of cell adhesion molecule E-cadherin and de-creased the wnt signal pathway molecule β-catenin in the MGC803 cells.However, it did not affect the expression and se-cretion of tumor metastasis-related proteins MMP9 and uPA.CONCLUSION:Overexpression of N24p55γinhibits cell ad-hesion by influencing the expression of E-cadherin and β-catenin in the MGC803 cells.