Objective To screen promoter binding protein of cyclin B2 by using human liver cDNA library, and investigate the expression and regulation mechanism of cyclin B2 gene. Methods By using cyclin B2 biotinylated promoter DNA as the selective molecule, the T7select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment, and the DNA fragment was cloned into pGEM-Teasy vector. Results Sequence analysis was performed in 20 positive plaques, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 6 coding sequences were obtained, all of which were known ones. Conclusion Cyclin B2 promoter binding proteins were screened. The results will be useful for further study the expression and regulation mechanism of cyclin B2.

0.05).Conclusion Levofloxacin treating community-acquired pneumonia than gatifloxacin have more medicine economics advantage and it is in the prior choice.

0.05).Conclusion Gatifloxation is safe and efficient in the treatment of community acquired pneomonia.

Objective To clone and identify human genes transactivated by homo sapiens HCV FTP2 by constructing a cDNA subtractive library with suppression subtractive hybridization tech- nique.Methods Suppression subtractive hybridization(SSH)and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV FTP2.The mRNA was iso- lated from HepG2 cells transfected pcDNA3.1(-)-HCV FTP2 and pcDNA3.1(-)empty vector re- spectively,and SSH method was employed to analyze the differentially expressed DNA sequence be- tween the two groups.After digestion with restriction enzyme Rsa I,small-size cDNAs were ob- tained.Then tester cDNA was divided into two groups and ligated to the specific adaptor I and adap- tor 2 respectively.After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR and then was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5?.Futhermore,the cDNA was se- quenced and analyzed in GenBank with Blast search after PCR.Results The subtractive library of genes transactivated by HCV FTP2 was constructed successfully.The amplified library contains 71 positive clones.Colony PCR shows that 56 clones contain 200～1000 hp inserts.Sequence analysis was performed in 24 clones randomly,and the full length sequences were obtained with bioinformatics method.Altogether 20 coding sequences in total were obtained,consisting of 19 known and 1 un- known.Conclusion The obtained sequences may be target genes transactivated by HCV FTP2,and some genes coding proteins involved in cell cycle regulation,metabolism and cell apoptosis.

Ascites ; complications ; etiology ; Heart Failure ; complications ; etiology ; Hemochromatosis ; complications ; diagnosis ; Humans ; Middle Aged

Objective To study the changes and significance of the frequencies of circulating follicular helper T cells (cTfh) and circulating regulatory follicular T cells (cTfr) as well as the cTfh/cTfr ratio in neuromyelitis optica spectrum disorder (NMOSD).Methods The frequencies of cTfh,cTfr and B cells in patients with NMOSD and health controls(HCs) were measured by flow cytometry.Enzyme-linked immunosorbent assay was used to detect the level of IL-21 and AQP4-Ab in patients and HCs.Results The frequencies of cTfh and B cells,the cTfh/cTfr ratio and the plasma level of IL-21 werc significantly higher in the relapsing patients than those in the remitting patients and HCs(P ＜ 0.05),and the cTfr level in the relapsing patients was lower than that in the remitting patients and healthy population (P ＜ 0.05).But no statistical differences were observed in the above indexes between the remitting paticnts and HCs.There was also no significant difference in AQP4-Ab level between the patients with relapse and remission (P ＞ 0.05).The frequency of cTfh in the patients wasc positively correlated with the level of B cells and IL-21(P ＜ 0.05),and the frequency of cTfr was negatively correlated with B cells and IL-21 (P ＜ 0.05).The ratio of cTfh/cTfr was positively correlated with B cell frequency and IL-21 level (P ＜ 0.05).AQP4-Ab level had no correlation with the frequencies of cTfh cells and B cells,cTfh/cTfr ratio and IL-21 concentration (P ＞ 0.05).Conclusion The changes in the frequencies of cTfh and cTfr as well as the imbalanced cTfh/cTfr ratio may promote the activation of humoral immunein NMOSD and participate in the pathogenesis of this disease.

Animals ; Extremities ; blood supply ; Ischemia ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Nitric Oxide ; blood ; Oxidative Stress ; Rats ; Rats, Wistar

Acute myocardial infarction (AMI) is one of the most common cardiovascular diseases. The clinical pathway is the therapeutic program for disease-specific treatment and its implementation may reduce both the duration and cost of the hospital stay. This study aims to construct and evaluate the efficacy of clinical pathways (CPs) based on integrated traditional Chinese and Western medicine for patients with AMI.

Objective To compare the expression changes of neuropeptide Y (NPY) receptor Y1 in different stages of osteoblast differentiation of rat bone marrow mesenchymal stem cells (BMSCs).Methods The rBMSCs were isolated in vitro from Sprague-Dawley (SD) rats using whole bone marrow adherence method and cultured. Then, the rBMSCs were divided into osteoblast-induced group and noninduced group. In different periods of culture at 1, 2 and 3 weeks, identification of the osteoblasts was performed by using immunocytochemistry and Western blot. Expressions of mRNA and protein of Y1 receptor were detected by real time reserve transcriptase-polymerase chain reaction (RT-PCR) and Western blot. Results RT-PCR demonstrated that osteoblast-induced group had a lower expression of Y1 receptor than non-induced group at the same time point and the expression of Y1 receptor was increased in a time-dependent manner in both groups. Western blot demonstrated higher expression of Y1 receptor in osteoblast-induced group compared with non-induced group at the same time point and a decreased expression of Y1 receptor in a time-dependent manner in both groups. Conclusions During the process of osteoblastic differentiation of rat BMSCs, the expressions of mRNA and protein of NPY Y1 receptor show different trends, when NPY may mediate the inhibition of osteoblastic differentiation of BMSCs through Y1 receptor pathway.

Objective To investigate the effects and mechanism of calcitonin gene-related peptide (CGRP) and substance P (SP) on proliferation of rat bone marrow mesenchymal stem cells. Methods The rBMSCs were isolated using whole bone marrow adherence method. In the different periods of culturing (1, 2,and 3 weeks), expressions of the neuropeptide receptors were detected by Western Blot and reserve transcriptase-polymerase chain reaction (RT-PCR). The BMSCs were treated with CGRP and SP at concentration 10-8 mol/L at different time (1,3,5,7,9 days), cell proliferation was detected with MTT assay, the protein expressions of cyclin D1 ,cyclin E and p53 were examined using Western Blot. Results The CGRP receptor and SP receptor were expressed in BMSCs. The expression of CGRP receptor was statistically higher than that of SP receptorat the same time point. The growth curves of BMSCs cultured by both neuropeptides had similar appearance. CGRP and SP stimulated the proliferation of BMSCs significantly at 9 days and 7 and 9 days. In this process, the expressions of cyclinDl and cyclinE were up-regulated by CGRP, SP only enhanced the expression of cyclinE; these effects all reached a peak at 5 days. The expression of p53 was down-regulated by both neuropeptides. Conclusion CGRP and SP had direct effects on the proliferation of BMSCs, the regulation of cell cycle proteins is one of the mechanisms.