1.APS on Inducing the Effect of Differentiating Cord Blood Monocyte into the Dendritic Cells Induced by Astragalus Polysaccharides In Vitro and its Immunological Characteristic.
Min DENG ; Xiaobing DOU ; Yiqian SHI
Journal of Medical Research 2006;0(09):-
Objectives To elucidate the effect of APS replacing cytokine on inducing the cord blood monocyte in vitro into the dendritic cells (DCs) and its cellar immunological characteristic. Methods The cord blood monocytes were isolated and obtained by lymphocyte isolation. three groups were divided: ②Cultured in the RPMI-1640 culture with GM-CSF/IL-4/TNF-?,as the positive control group, with APS in concentration (100mg/L) as the experimental group,and without GM-CSF/IL-4/ TNF-?and APS,as the negative control group, respectively. The morphotype of DCs was identified by inverted optical microscope or scanning electron microscope. The phenotype of cultured 12 days DCs (CD1a, CD80, CD86, and CD83) was identified by flowcytometry. Results Cultured for 72 hours , the morphous of cell of the experimental group grew clustering and began to change from round to irregularity, appearing rough cell face and barb pustute. The longer cell cultured, the more obvious the dendritic structure is. The experimental group cell cultured for 12 days had the most typical dendritic structure. the negative control group cell had no dendritic structure and became the macrophage when cultured for 12 days. The experimental group cell cultured for 10 days showed typical dendritic morphotype by SEM. The experiment group cell and the positive group cell cultured for 12 days significantly expressed the high level phenotype of DCs((CD1a, CD80, CD86, and CD83))by flowcytometry. Conclusions APS and cytokine both could induce the cord blood monocyte to direofive differentiate into functional DC.
2.Effect of estrogen replacement therapy on chemokine receptor CXCR2 in monocytes in postmenopausal women with coronary artery disease
Zhubin LEI ; Jianping LIU ; Min WEI ; Jingbin DOU ;
Academic Journal of Second Military Medical University 1999;0(12):-
Objective: To study the change of chemokine receptor CXCR2 in monocytes in postmenopausal women with coronary artery disease (CAD) after estrogen replacement therapy. Methods: Randomized placebo controlled trial was conducted in 22 post menopausal women with CAD and 20 normal menopausal women by giving 0.625 mg premarin daily or vitamin C treatment for 3 months. Serum estradiol (E 2) and chemokine receptor CXCR2 in peripheral blood monocytes were measured at 0 and 3 months of treatment. Results:(1) E 2 and CXCR2 levels in the CAD group was lower than that in the normal group ( P
3.Application of modified double-lumen balloon catheter for patients with achalasia of the cricopharyngeus muscle
Ling MENG ; Min LU ; Zulin DOU ; Qiaoling LI ; Jun PENG
Chinese Journal of Nursing 2010;45(4):304-306
Objective To evaluate the feasibility and safety of double-lumen balloon catheter applied in patients with achalasia of cricopharyngeal muscle. Method Fifty patients with achalasia of cricopharyngeal muscle were randomly divided into experimental group and control group. All the patients received routine drug treatment,swallowing function training,feeding training and low frequency VitalStim electric stimulation. In addition,double-lumen balloon catheter and #14 urinary catheters were applied to patients in the experimental group and control group,respectively. The swallow water tests and video fluoroscopy swallowing study(VFSS) were used to evaluate the treatment effects,the electron-nasopharyngolaryngoscope was used to assess bleeding and swelling of mucous membrane,and VRS-5 was used to assess pain. Result After treatment,the scores of swallow water tests and VFSS were significantly better than those before treatment in both groups(P<0.05). There was no significant difference between the two groups(P>0.05). However,the incidence of complications was significantly higher in the control group than that of experimental group(P<0.05). Conclusion Both treatment methods can effectively relieve the achalasia of cricopharyngeal muscle,but modified double-lumen balloon catheter can reduce the incidence of complications.
4.Astragalus polysaccharides(APS) induces the cord blood monocytes into mature dendritic cells(DCs) directly and its impact on T cells proliferation
Min DENG ; Xiaobing DOU ; Yiqian SHI ; Xingde WO
Chinese Journal of Immunology 1985;0(06):-
Objective:To observe the effect of Astragalus polysaccharides(APS) on inducing the cord blood monocytes into mature dendritic cells(DCs) in vitro and to investigate their morphous,cellar immunological characteristics,and contribution to T cell proliferation.Methods:①The cord blood monocytes were isolated by lymphocyte isolating solution under axenic condition,and three groups were divided.②Cells cultured with APS in concentration of 100 mg/L as the experiment group, that with the cytokines of GM-CSF/IL-4/TNF-? as the positive control, and another without either GM-CSF/IL-4/TNF-? or APS as the negative control, respectively. The morphology of DCs was identified by inverted optical microscope or scanning electron microscope. The phenotype of 12 days cultures of DCs(CD1a, CD80, CD86 and CD83) were identified by flowcytometry. The DCs preparations from the experiment group were treated with mitomycin for 45 min to remove their proliferative activity as incentive cells in the mixed cultures with allogenic peripheral blood mononuclear cells from healthy volunteers as responders. T cell proliferation induced with the DCs preparations was detected by MTT chromatometry.Results:After cultured for 72 hours, the cell of both the experiment group and the positive control grew dusteringly and began to change from round to irregularin shapes. The longer the cells were cultured, the more obvious the dendritic structure is. The cells of experiment group and the positive control group when cultured for 12 days had the most typical dendritic structure. The negative control group cells had no dendritic structure and became macrophages when cultured for 12 days. The experiment group cells cultured for 10 days showed typical dendritic morphology by SEM. The experiment group cells and the positive group cells cultured for 12 days significantly expressed high level of the phenotypes of DCs(CD1a, CD80, CD86 and CD83) by flowcytometry.And the difference exhibitied statistical significance when compared with the negative control group(P0.05).The mixed lymphocyte reaction showed that the DCs induced by APS trigerred proliferation of allogenic T cells obviously.Conclusion:Both APS and cytokine could induce the cord blood monocytes to differentiate into functional DCs committedly. DCs reduced by APS stimualate proliferation of the allogenic T cells obviously.
7.In vitro anti-metastatic role of miR-10b through modulation of cytoskeleton in human hepatocellular carcinoma cell
Qingjun LI ; Jinxue ZHOU ; Feng HAN ; Min ZHANG ; Xianzhou ZHANG ; Kefeng DOU
Chinese Journal of Hepatobiliary Surgery 2015;21(3):194-199
Objective To investigate the effect of miR-10b on migration and invasion of human hepatocellular carcinoma (HCC) cell lines.Methods Transwell assay was used to evaluate the motility and invasiveness in different HCC cell lines,qRT-PCR was then used to detect the expression levels of miR-10b in different HCC cell lines.Artificial mimics of miR-10b were transiently transfected into HepG2 cells,which have low expression of miR-10b,and then the changes in migration and invasion were evaluated by Transwell assay.Antagomirs of miR-10b (miR-10b-AS) were transiently transfected into MHCC97H cells,which have high expression of miR-10b,and then the changes in migration and invasion were evaluated as well.Cell morphology changes were detected by immunofluorescence and electron microscopy,Results There was a significant correlation of miR-10b expression level with cell motility and invasiveness.Low-level expression of miR-10b was observed in HepG2 cells,which exhibited weak motility and invasiveness; whereas high-level expression of miR-10b was observed in MHCC97H cells,which exhibited strong motility and invasiveness.Up-regulation of miR-10b expression in HepG2 cells increased cell motility and invasiveness,whereas inhibition of miR-10b reduced cell motility and invasiveness in MHCC97H cells.Immunofluorescence and electron microscopy results showed that up-regulation of miR-10b in HepG2 cells significantly increased proliferation of filopodia and lamellipodia,whereas inhibition of miR-10b decreased filopodia and lamellipodia amount in MHCC97H cells.Conclusion miR-10b is involved in the invasion and metastasis of HCC cell through regulation of cytoskeleton in vitro and inhibition of miR-10b is likely to be a new molecular target to block metastasis.
8.Construction of the Expression Vector of Viruslike Particles Containing FMDV IRES RNA
Min DOU ; GuoGuang ZHANG ; GuangFu YU ; HongXin ZHANG ; MingShan SHEN ; Liang CHEN ;
China Biotechnology 2006;0(09):-
The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.
9.Expression of myosin light chain kinase in acute lung injury
jin-shi, LI ; jian-wen, BAI ; min-jia, LIN ; dou-xia, ZHANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(12):-
Objective To study the inflammation and expression of myosin light chain kinase(MLCK) through establishing acute lung injury animal model of mice induced by lipopolysacchride(LPS), and approach the role of MLCK in the mechanism of acute lung injury.Methods Twenty female BALB/c mice were randomly divided into LPS group(n=10) and control group(n=10).The BALB/c mice of LPS and control groups were induced by 30 ?L 0.9% NaCl via intranasal instillation,while only LPS group was treated with LPS(20 ?g/each mice).The pathology,wet/dry lung weight ratio and the total cell quantitation in bronchoalveolar lavage fluid(BALF)were compared between these two groups.Furthermore,immunohistochemistry assays were used to determine the status of MLCK expression in the lung.And RT-PCR was adopted to determine the status of MLCKmRNA in the lung. Results Compared with the control group,the LPS group showed more serious pulmonary hemorrhage,edema and infiltration of neutrophils, significantly increased water content in the lungs and total cell quantitation in BALF(P
10.Effect of montelukast and dexamethasone on inflammation of asthma
min-jia, LIN ; jian-wen, BAI ; dou-xia, ZHANG ; jin-shi, LI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(01):-
Objective To study the effect of montelukast(MK)and dexamethasone(Dex)on inflammation of asthma.Methods Asthma model was established and treated with MK or Dex.Bronchoalveolar lavage fluid(BALF) and histopathologic change were observed,IL-5mRNA of lung and bone marrow cells were detected by in situ hybridization,IL-5 immunoreactive cells by immunohistochemistry,and CD34+ and CD3+ of bone marrow cells by flow cytometry. ResultsCompared with asthma group,the number of total cells and eosinophils in BALF of MK group and Dex group were significantly decreased(P0.05). Conclusion MK and Dex can well inhibit airway inflammation and expression of IL-5mRNA in lung and bone marrow cells,though MK may be inferior to Dex in some aspects.The combined treatment of leukotriene receptor antagonist and glucocorticosteroid may be a new direction for asthma.