Apolipoproteins A ; therapeutic use ; Coronary Disease ; drug therapy ; prevention & control ; Humans ; Recombinant Proteins ; therapeutic use

Desensitization, Immunologic ; methods ; Oligodeoxyribonucleotides ; therapeutic use ; Toll-Like Receptor 9 ; therapeutic use

A new method was developed for the separation and determination of inosine monophosphate (IMP) by ion chromatography (IC) with suppressed conductivity detection. Separation was achieved on an anion-exchange column Ionpac AS11-HC of high capacity within a short time. 30 mmol/L KOH produced by an EGC-KOH eluent generator was used for isocratic elution. No interferences existed between the seven common inorganic anions and IMP. Under the optimum conditions, the linear range of the calibration curve for IMP was 1. 0-200 mg/L, with correlation coefficient ( R2 ) of 0. 999. The relative standard deviations (RSDs) for retention time, peak height and peak area of IMP were 0. 16%, 0. 94% and 0. 86%, respectively, indicating good reproducibility of the method. The method was successfully applied to the determination of IMP in meat products, with spiked recovery ranging from 86. 0% to 110. 0%. This simple, accurate and reliable method could be served as a rapid and effective analytical tool for meat flavoring research.

Objective To explore the changed status of the intestinal flora microecology in the model infant rats with irritable bowel syndrome(IBS) by integrative method.Methods To establish 20 infant rats model with IBS.The IBS rats were randomly divided into the treatment group A with bifico and positive control group B,and another 10 infant rats which grew up regularly were taken as negative control group C.The feces smear staining and intestinal flora detection was performed in the 3 groups respectively,meanwhile the dry and wet weight of each group rats feces and the content of water in the feces were compared.Results The intestinal flora imbalance rate of infant rats IBS group(A+B) was 70%,and there was 30% in the control group C,the difference between model group and group C was very significantly(?~2=4.34 P

In order to study the correlation between microbial diversity and the pollution degrees of the ruraldrinking water in Dongjiang River basin. Five types of drinking water of this basin were collected,and fifteen water samples of five types of drinking water of this basin had been collected from reservoir,centralized water supply wells,wells in the vicinity of pig farms,wells nearby embankment and wells in villages. The six(physical,chemical,and biological) property indices of water samples were tested,at the same time,the DGGE analysis was done. The results of PCR-DGGE fingerprint indicated that bacterial richness of these drinking water samples were high,and different samples in fingerprint were different distinctively. The UPGMA dendrogram of sample basis on DGGE fingerprints showed the structure of different types of bacteria in drinking water in rural communities is obvious differences. And the results of CCA showed that the concentration of phosphorous has the largest relevance to the community structure of bacteria in water samples,followed by the concentration of nitrogen in the water. Ten typical bands were excised and sequenced. The sequences obtained were affiliated with Spirochaetes,Cyanobacteria,Proteobacteria,Actinobacteria,Acidobacteria.

Objective To investigate the effects of ischemic postconditioning (Post) on myocardial cell apoptosis and expres- sion of Bcl-2 and Bax protein during ischemia/reperfusion period in rabbits.Methods Eighteen rabbits were randomly allocated to three groups (6 in each group),sham operation (group S),ischemia/reperfusion group(group IR) and ischemic postconditioning group(group Post).Group IR and group Post were subjected to 15 minutes of left anterior descending coronary artery occlusion followed for 30 minutes of reperfusion.Ischemic postconditioning was achieved by three 30 seconds cycles of reperfusion,each followed by 30 seconds ischemia.Cardiomyocyte apoptosis were determined by in situ TDT-mediated dUTP nick end labeling (TUNEL) and DNA electrophoresis.The expression of Bcl-2 and Bax proteins in apoptotic myocardial cells were detected by immunohistochemistry sepa- rately.Results Compared with group IR,apoptotic index was significantly reduced in group Post [(28.06?2.92) % vs.(55.70? 13.96)%,P

Objective　Recombinant human pro-urokinase forms insoluble inclusion body when overexpressed in Escherichia coli. It must be denatured and renatured in vitro so that it can acquire activity. This study aimed at increasing the renaturation yield of denaturant pro-urokinase. Methods　We evaluated the basic renaturation conditions of pro-urokinase through qualitative and quantitative analysis of pH, temperature, denatured concentration, protein concentration, and the ratio of reduced and oxidized thiol reagents. We also compared the effects of nonspecific additives, step-wise dilution and urea gradient dialysis. Results　We defined the optimal conditions of pro-urokinase renaturation with a yield of about 20%-30%. Conclusion　Different recombinant denatured proteins have different renaturation conditions due to their different molecular sizes, molecular constructions, disulfide bond numbers, and hydrophobicity. The renaturation yield can be increased by optimizing the renaturation conditions of a specific protein.

To examine the effects of heterologous expression of ZrGPD1 (encoding glycerol 3-phosphate dehydrogenase ) cloned from osmotolerant yeast Zygosacharomyces rouxii on glycerol production in wild Pichia farinosa,the URA3 gene was amplified from P. farinosa as the homology integrative region. A recombinant plasmid (pUR-ZG) was constructed then transformed into P. farinosa by electroporation. The transformant pfa-gu was obtained by the selectable marker Zeocin TM . Primary results showed that the biomass of pfa-gu was higher than the wild type in the flask and after 72h fermentation the concentration of glycerol of pfa-gu was 37g/L enhanced 30% in comparison with the wild type. It is concluded that heterologous expression of ZrGPD1 is useful for increasing glycerol production and the ability of osmoregulation in P. farinosa.

The ?glutamyltranspeptidase encoding gene(ggt) from Bacillus subtilis SYU 20016 was amplified by PCR. The ggt gene was inserted in pBV220 to yield the recombinant expression vector pBV220ggt. Overexpression of ggt in E.coli JM109 was achieved with pBV220ggt. SDSPAGE analysis showed an overexpressed recombinant product at about 65kDa,consistent with the molecular weight predicted from gene sequence. The ferment conditions of r-glutamyltranspeptidase were also discussed. The optimum temperature and pH for the enzyme were determined as 30℃ and 7.2 respectively.The cultures were incubated at 42℃ for 4h with broth volume 20ml/250ml flask and the yield of 6U/ml was obtained, enzyme activity of B. subtilis NX2 was only 3.2 U/ml.

C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.