Animals ; Cells ; metabolism ; radiation effects ; Electromagnetic Fields ; Humans ; Reactive Oxygen Species ; metabolism

Objective:To investigate the impact of recombinant flagellin targeting TLR5 and NLRC4 simultaneously or respectively on innate immune cells in mice. Methods: Induction,expression,purification and identification of recombiant FliC,which were FliC(activating both TLR5 and NLRC4);FliCΔ90-97(unable to activate TLR5),FliC-L3A(unable to activate NLRC4),FliCΔ90-97:L3A(unable to activate both TLR5 and NLRC4). The mice were divided into five groups,namely group FliC,FliC-L3A,FliCΔ90-97,FliCΔ90-97:L3A and PBS,which were injected with 100μl PBS or 10μg recombinant flagellin intraperitoneally,three mice in each group. 12 h later,the mice were executed using dislocation of cervical vertebra and the splenic and peritoneal cells were isolated. The spleen was grinded into single-cell suspension. The proportion of neutrophils,NK cells,DCs and the expression level of CD80 and CD86 on DCs were evaluated with flow cytometry. Results:Group FliC,group FliC-L3A and group FliCΔ90-97 shared the similar proportion of neutrophils in peritoneal cavity ( P>0. 05 ) , and all of which were significantly higher than group PBS and group FliCΔ90-97 ( P<0. 01),and NK cells also showed the similar trend. Compared with group FliCΔ90-97 and FliCΔ90-97:L3A,the mean fluorescence intensities(MFIs) of CD80 and CD86 in group FliC and FliC-L3A increased significantly(P<0. 01). The proportion of Treg in spleen was highest among all groups. Conclusion:Activation of TLR5 and NLRC4 had similar chemotaxis of neutrophils and NK cells. The ex-pression of CD80 and CD86 on DCs were upregulated after stimulation by flagellin and TLR5-dependent. Activation of TLR5,but not NLRC4,increased the proportion of Treg in spleen.

Objective To detect the distribution of site-174G/C,site-572C/G and site-597G/A of interleukin-6 gene promoter regions in patients with endometriosis,and to analyze polymorphism of the promoter.Methods Interleukin-6 gene promoter region polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism in patients with endometriosis(n=98)and normal controls(n=99).Among the 98 patients with endometriosis,22 were defined as stageⅡ,35 as stage Ⅲ and 41 as stage Ⅳ.Comparisons between the two groups on the genotype of IL-6 gene promoter and the distributed allele frequency were done,and also among the patients of stage Ⅱ,Ⅲ and Ⅳ once significant differences were detected in genotype frequency or allele frequency in disease group and control group.Results At the site-174,the frequencies of GG,GC and CC genotypes were 100%,0% and 0%,respectively,in patients with endometriosis,while 98.99%,1.01% and 0%,respectively,in normal controls.No significant difference was found in the distribution frequency of IL-6 genotype between patients with endometriosis and normal controls.At the site-572,the frequencies of CC,CG and GG genotypes were 55.10%,42.86% and 2.04%,respectively,in patients with endometriosis,while they were 70.71 %,28.28% and 1.01%,respectively,in normal controls.A significant difference existed between the two groups.At the site-597 of IL-6 gene,no GA and AA genotypes were found in the two groups.At the site-572,the frequencies of CC,CG and GG genotypes were 59.09%,40.90% and 0%,respectively,in patients with endometriosis of stage Ⅱ,54.29%,42.86% and 2.85% in patients of stage Ⅲ,and 53.66%,43.90% and 2.44% in patients of stage Ⅳ.No significant difference was found among the three groups.Conclusion Mutations at the sites-174 and-597 of IL-6 gene promoter region were rare.Their distribution frequencies have nothing to do with morbidity of endometriosis.While at the site-572,its distribution frequency of genotype shows correlation with morbidity of endometriosis but does not show correlation with the extent of endometriosis.

Objective To construct prokaryotic expression vector of NS5A transactivating protein 7 of hepatitis C virus (NS5ATP7), induce the expression of NS5ATP7 in E. coli, and to predict its structure and function by bioinformatics analysis. Methods NS5ATP7 gene was amplified by reverse transcription PCR (RT-PCR) using HepG2 cells mRNA as template, and was ligated into pGEM-T cloning vector. After verifying the sequence of the inserted fragment by restriction enzyme digestion identification and sequencing, NS5ATP7 was cloned into inducible prokaryotic expression vector pET-32a(+) and transfected into competent E. coli BL21. The protein expression was induced with IPTG and the product was analyzed by SDS-PAGE and Western blotting hybridization. The structure and function of NS5ATP7 were predicted using bioinformatics techniques. Results NS5ATP7 gene with about 891bp was amplified by RT-PCR, which was coincident with that we expected, and it was then inserted into expression vector pET-32a(+). Under the induction of IPTG, the recombinant NS5ATP7 was expressed successfully. SDS-PAGE and Western blotting assay showed that this recombinant protein possessed good immunogenicity. Bioinformatics method such as ExPASy, TMHMM and SignalP analysis indicated that NS5ATP7 was full of helices, had neither transmembranous structure nor signal peptide. Conclusions The recombinant NS5ATP7 gene could be successfully expressed in prokaryotic expression system of E. coli, which might lay the foundation of studying the immunogenicity and biological charactersitics of the NS5ATP7. Bioinformatics analysis may provide a new method to analyze the structure and function of a new protein.

Objective To investigate the status of personality disorder and the relationship with childhood trauma in heroin addicts.Methods 108 heroin dependent patients were investigated by Personality Disorder Ques-tionnaire -4 +(PDQ -4 +)and Childhood Trauma Questionnaire -Short From (CTQ -SF).Results The comor-bidity rate of heroin dependence and personality disorder was between 47.2% and 10.2%,which had a higher posi-tive rate of for borderline (47.2%)and antisocial (38.0%)personality disorder and the lowest for depression (10.2%).The borderline personality disorder score was highest[(4.63 ±2.16)points]and the lowest was histrionic personality disorder [(2.65 ±1.49)points]in 12 subscales,which were significantly higher than those in the national norm,and the differences were statistically significant (t =13.54,2.57,all P <0.05).It had significantly positive correlation between borderline and antisocial personality disorder and the each five factors of childhood trauma experi-ence (r =0.265,0.390,0.207,0.303,0.276;0.225,0.219,0.228,0.236,0.287,all P <0.05 ).In addition, depressive personality disorder and physical abuse,emotional neglect and body ignored showed significantly positive correlation (r =0.226,0.227,0.196,all P <0.05).Conclusion There was the comorbidity rate of personality disorders and heroin dependence,and the higher positive rate was borderline and antisocial personality disorders.It was a certain correlation of childhood traumatic experience and personality disorders,especially borderline,antisocial and depressive personality disorder.

0.05) in VAS and ramsay scale, but side effects were significantly different in group B compared with those in group S. Conclusion: The analgesia efficacy of S is same as B in maxillofacial postoperation, but side effects are more significantly in group B, and group S is more safe and handy.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hepatitis B ; epidemiology ; immunology ; prevention & control ; Humans ; Infant ; Infant, Newborn ; Male

Objective To explore the effects of earlier oral-motor exercise on swallowing and feeding capability of children with cerebral palsy. Methods 36 children accepted oral-motor exercise performed by speech-language therapist once a day for 3 months. They were assessed for oral motor and feeding before and after treatment, and compared with other 36 controls matched with age and type. Results The oral motor in treatment group improved significantly （t=2.184,P<0.05） after treatment, but merely in control. The difference of scores of oral motor before and after treatment was (9.89±4.33) in treatment group, and (3.38±1.56) in control group (t=7.423, P<0.01). Conclusion Early oral-motor exercises is effective to improve oral motor function and feeding capability.

Acquired Immunodeficiency Syndrome ; prevention & control ; China ; epidemiology ; Condoms ; utilization ; Humans ; Surveys and Questionnaires ; Transients and Migrants

OBJECTIVEEndothelial apoptosis plays an important role in the initiation of atherosclerosis. It would be useful to clarify whether activation of non-neuronal muscarinic receptor (NNMR) could prevent endothelial apoptosis and atherosclerosis. We investigated the effects of NNMR activation on regulating rat aortic endothelial cells (RAECs) apoptosis induced by homocysteine, an independent risk factor of atherosclerosis, and further studied its molecular mechanism.

METHODSRAECs were incubated using homocysteine at the concentration of 2.7 mmol/L for 36 h. RAECs were also pre-treated with carbachol or arecoline to examine their effects. RT-PCR was used to assess changes in the gene expression related to cell apoptosis.

RESULTSIncubation of RAECs with homocysteine at the concentration of 2.7 mmol/L resulted in morphologic changes, such as cellular shrinkage, membrane blebbing, chromatin condensation and margination. These could be attenuated by pretreatment with carbachol and arecoline at the concentration of 10 micromol/L for 12 h. Homocysteine induced apoptosis in RAECs and the molecular mechanisms were associated with the regulation of fas, fas-L and caspase-8 in the death receptor pathway, bcl-2, bcl-xL and bax in the mitochondrial pathway, caspase-12 in the endoplasmic reticulum pathway and caspase-3, caspase-6 and p53 as downstream effectors. Carbachol and arecoline attenuated the effects of homocysteine on genes in the death receptor pathway, in the mitochondrial pathway and in the downstream pathway. Atropine could reverse all of the effects of arecoline.

CONCLUSIONActivation of NNMR by carbacol and arecoline inhibits homocysteine-induced endothelial cell apoptosis mainly through regulation of death receptor pathway, mitochondrial pathway and downstream effectors.

Animals ; Aorta ; cytology ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Arecoline ; Carbachol ; Cell Cycle ; Endoplasmic Reticulum ; metabolism ; Endothelial Cells ; cytology ; drug effects ; Homocysteine ; adverse effects ; Mitochondria ; metabolism ; Rats ; Receptors, Muscarinic ; metabolism