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.
		                        		
		                        		
		                        		
		                        	
4.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.
		                        		
		                        		
		                        		
		                        	
5.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
		                        		
		                        		
		                        		
		                        	
6.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.The correlation between atypical respiratory pathogens infection and serum total IgE levels
Hongjing LIU ; Min DOU ; Hui CHEN ; Jin CHEN ; Xiujun PAN ; Lisong SHEN
International Journal of Laboratory Medicine 2015;(17):2480-2481,2484
		                        		
		                        			
		                        			Objective To investigate the correlation between atypical respiratory pathogens infection and serum total IgE levels . Methods Serum IgM level was detected in 1 913 blood samples of children with atypical respiratory infection by using indirect im‐munofluorescence assay ,including mycoplasma pneumonia(MP) ,legionella pneumophila (LP) ,rickettsia Q(QFR) ,chlamydia pneu‐monia(CPn) ,adenovirus (Adv) ,respiratory syncytial virus (RSV) ,influenza A virus (IAv) ,influenza B virus (IBv) and parainflu‐enza virus (PIV)1/2/3 .The serum total IgE level was detected by immune scatter turbidimetry .Software SPSS 17 .0 was used in data statistical analysis .Results A total of 991 out of 1 913 samples of respiratory inflected children exhibited positive(positive group) ,while 922 exhibited negative(negative group) in indirect immunofluorescence assay .650 out of the 991 positive samples (65 .59% ) contained MP infection and the combination of MP infection and other virus infections .The serum total IgE level in posi‐tive group was significantly higher than that of the negative group ,and the serum total IgE level in samples with MP infection was higher than that in samples with IBv infection ,Adv infection ,and RSV infection .In the samples in which serum total IgE level was higher than the clinical reference range (100 kU/mL) ,the infection rate of MP infection alone was 31 .29% ,which was evidently higher than that in samples of low IgE level(< 100 kU/mL ,21 .30% ) .On the other hand ,the infection rate of RSV alone was 1 .88% and the infection rate of Adv alone was 3 .13% ,which were both evidently lower than those in samples with normal serum total IgE level(both 6 .53% ) .Conclusion MP is the most common pathogen in children with atypical respiratory pathogen infec‐tion ,and can lead to higher serum total IgE levels .
		                        		
		                        		
		                        		
		                        	
8.Role and mechanism of microRNA-15b in the regulation of epithelial-mesenchymal transition of peritoneal mesothelial cells
Jiayi CHEN ; Haitang HU ; Jianyi PAN ; Wei ZHANG ; Jinzhong CHEN ; Shaoxin ZHONG ; Min MO ; Xianrui DOU
Chinese Journal of Nephrology 2017;33(4):290-295
		                        		
		                        			
		                        			Objective To explore the role and mechanism of microRNA-15b in the regulation of epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs).Methods PCR assay was used to determine the expression of microRNA-15b in the HMrSV5 induced by 138mmol/L high glucose for 24 h.MicmRNA-15b mimic or inhibitor was transfected into human peritoneal mesothelial cells (HMrSV5) to over-express or down-regulate microRNA-15b.The cells were then incubated with 138 mmol/L high glucose for 24 h,and the expressions of E-cadherin(E-Cad),Vimentin (VIM),Fibronectin(FN) and Smad7 were detected by real-time PCR and Western blotting respectively.Results microRNA-15b in the HMrSV5 ceils was over-expressed and down-regulated.Increased level of microRNA-15b was obtained in HMrSV5 cells treated with high glucose.In vitro,high glucose led to the up-regulation of vimentin as well as fibronectin and the down-regulation of E-cadherin in HMrSV5 cells (all P < 0.05),which indicated EMT and fibrosis.Suppression of microRNA-15b by transfection with microRNA-15b inhibitor partially reversed the EMT and fibrosis changes (P < 0.05),while over-expression of microRNA-15b by transfection with microRNA-15b mimic obviously enhanced the EMT and fibrosis changes (P < 0.05).Conclusions MicroRNA-15b mediates high glucose induced EMT in human peritoneal mesothelial cells by the inhibition of Smad7 possibly.MicroRNA-15b maybe a new target for the prevention and treatment of peritoneal fibrosis during peritoneal dialysis (PD).
		                        		
		                        		
		                        		
		                        	
9.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.
		                        		
		                        		
		                        		
		                        	
10.Comparison of Rhizospheric Soil DNA Isolation and Purification Methods from the Root of Fritillaria thunbergii Miq
Ying-Ying DOU ; Zhan-Min LIN ; Ying-De ZHU ; Qun LU ; Bo-Ping YE ;
Microbiology 2008;0(11):-
		                        		
		                        			
		                        			Six DNA extraction methods and four DNA purification methods were compared and analyzed in this study to get higher quality DNA from the rhizospheric soil of Fritillaria thunbergii Miq.Results showed that higher purity DNA were harvested by pretreating the soil with 20 mmol/L EDTA(pH 7.5),then isolating soil DNA with CTAB-SDS-frozen-thawing,and further purified by agarose method.The recovery rate of this soil DNA was about 44.00 ?g/g ? 2.65 ?g/g soil,and they were qualified for the microbial diversity analysis in the rhizospheric soil of F.thunbergii Miq based on the 16S rDNA sequence.
		                        		
		                        		
		                        		
		                        	
 
            
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