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.

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.

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

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 observe and compare the different forces between doctors and nurses used visible laryngoscope endotracheal intubation applied to the oropharyngeal organization. Methods 10 nurses (to carry on laryngoscope intubation theory, and had certain study period practice) were chosen in group A and 10 clinical anaesthetize doctors (to be possible correctly used visible laryngoscopes) were chosen in group B, two groups used the visible laryngoscope on the same model person body inserted the tube, computer monitor software recorded results. Results The impulse force was (25.57±3.37) N·s and insert tube time was (25.3±3.3) s in group A which were higher than (16.47±2.99) N·s and (16.2±3.0) s in group B (t=2.550 and 2.207, P<0.05). The average forces in group A and group B were (0.87±0.62) N and (0.64±0.30) N, and peak forces were (3.05±0.95) N and (2.06±0.48) N, there was no remarkable difference between the two groups (P>0.05). Conclusions There is no statistics difference forces applied to the oropharyngeal organization between nurses and anaesthesiologists using visible laryngoscope intubation, and visible laryngoscope intubation technique is easy to learn and it is feasible by the nurse to master the technology and applied to anesthesia intubation care and emergency care.

OBJECTIVE:To select a technique for preparation of fluconazole for injection and to establish a method for determination of its content.METHODS:The formula was selected on the basis of pH value and species of solution adjuvant.The content was determined by HPLC.RESULTS:The preparation was stable when pH value was between6.5～8.5,and its clarity could be increased by propylene glycol.The detectable concentration of fluconazole showed a good linear correlation in the range of40～200?g/ml.The average recovery was100.37%,RSD=1.37%(n=3).CONCLUSION:The fluconazole for injection prepared by the present technique is stable in quality and the method for content determination is accurate and practicable.