Staphylococcus aureus (S. aureus) is an important human pathogen which can cause a chronic condition with a high relapse rate despite the aggressive antimicrobial treatment. Recent studies showed that intracellular pattern recognition receptors (including NOD) in response to bacteria or bacterial products play a proinflammatory role by activating nuclear transcription factor-κB (NF-κB). But how NOD2 mediates the proinflammatory response to S. aureus in mast cells (MCs) is unclear. So, in this study, we attempted to examine the role of NOD2 in inflammatory responses of MCs to S. aureus. P815 cells (a mouse mast cell line) were cultured. Real-time PCR was used to detect the NOD2 mRNA expression in P815 cells during S. aureus infection. The siRNA against NOD2 gene was synthesized and transfected into S. aureus-infected P815 cells. By using the methods of ELISA and flow cytometry, the effects of NOD2 gene silencing on cell phagocytosis, cytokine secretion, NF-κB activation and cell apoptosis of the S. aureus-infected P815 cells were examined. It was found that S. aureus infection could increase the expression of NOD2 mRNA in P815 cells. NOD2 gene interference in P815 cells reduced the number of S. aureus engulfed by P815 cells, the level of cytokines and the activation of NF-κB. In addition, S. aureus could induce the apoptosis of P815 cells, but NOD2 gene silencing did not affect the cell apoptosis rate. Our data suggested that NOD2 plays a key role in pathogen recognition, signal transduction, and NF-κB activation in the inflammatory responses of MCs infected by S. aureus.

Objective To investigate the effects of NOD2 on inflammatory responses of macrophages to Staphylococcus aureus. Methods Real-time RT-PCR detected NOD2 gene expression of macrophages infected by S. aureus. Synthesis of siRNA against NOD2 and interfere with macrophages, observed the effects of NOD2 gene silencing to phagocytosis of 5. aureus, cytokine secretion, activation of nuclear transcription factors, cell apoptosis of the macrophages infected by S. aureus using F.I .IS A, flow cytometry etc. Results S. aureus infection of macrophages can cause increased expression of intracellular NOD2. NOD2 gene silencing of macrophage lead to the decreased ability of phagocytosis with S. aureus, the lower levels of cytokines secretion, deficiencies of NF-κB activation. S. aureus can cause macrophage apoptosis, with the apoptosis rate increased with time. Conclusion The intracellular pattern recognition receptor NOD2 play a key role in pathogen recognition, signal transduction, activation of nuclear transcription factors in the process of macrophages infected by S. aureus.

3-Hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR), the first enzyme of mavalonic acid pathway, is one of the key devices involved in ginsenoside biosynthesis based on synthetic biology approach. The open reading frame of a novel HMGR gene from Panax ginseng (PgHMGR2) was cloned and analyzed in this study. PgHMGR2-encoding protein showed 71.6% sequence similarity to a P. ginseng HMGR in GenBank. The full-length cDNA sequence of PgHMGR2 containing 1 770 bp, which encodes 589 amino acids, was cloned by RT-PCR strategy from P. ginseng. The bioinformatic analysis showed that PgHMGR2-encoding protein contained two transmembrane regions and the HMG_CoA_reductase domain, without signal peptide. The protein sequence of PgHMGR2 had the highest sequence similarity (99%) with Panax quinquefolius HMGR (GenBank accession No. ACV65036). The expression level of PgHMGR2 was the highest in flower based on a real-time PCR analysis, followed by leaf and root, and the lowest was in stem. The result will provide a foundation for exploring the molecular function of PgHMGR2 involved in ginsenoside biosynthesis based on synthetic biology approach in P. ginseng plants.

Glycyrrhizin is the major bioactive compound in Glycyrrhiza uralensis. Farnesyl-diphosphate synthase (FPS) is one of the major enzymes involved in the glycyrrhizin biosynthetic. We cloned the full length of FPS coding sequence based on the transcriptome data of G. uralensis. The FPS gene of G. uralensis (GlyurFPS) is 1029 bp, coding 342 amino acid. The homology between the sequence of its protein and that of the FPS in Cicer arietinum, Medicago sativa, Medicago truncatula, Glycine max, Lotus japonicus was of 94%, 94%, 93%, 93% and 92%, respectively. Plant FPS gene is very conservative, therefore the phylogenetic tree is same with the plant classification consistent.

AIMTo detect the expression of a novel protein, hemangiopoietin (HAPO), in the human fetal liver at the protein level.

METHODSMonoclonal antibodies (moAbs) against HAPO were produced by traditional hybridoma technique. Their affinities were calculated from the results of non-competitive ELISA and the antibodies were purified by protein G affinity chromatography. Expression of HAPO in the liver was detected by SDS-PAGE and Western blot.

RESULTSFive strains of moAbs were screened out in total, among which three were IgG1 and the other two were IgM. Their light trains were all belonged to kappa. The relative affinities of the three IgG1 form moAbs were 3.06 x 10(9) mol/L, 6.07 x 10(8) mol/L and 1.71 x 10(10) mol/L respectively. After purification, the purity of the moAbs could reach more than 99%. HAPO expression was detected at the protein level in the human fetal liver, and the apparent molecular weight of the nature HAPO was very close to but a little higher than our recombinant one.

CONCLUSIONHAPO was expressed in the human fetal liver at the protein level.

Antibodies, Monoclonal ; Blotting, Western ; Fetus ; Humans ; Immunoblotting ; Liver ; embryology ; metabolism ; Proteoglycans ; metabolism

Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.
Codon ; Escherichia coli ; Genome, Fungal ; Reishi ; genetics ; Saccharomyces cerevisiae ; Transcriptome

Animals ; Dose-Response Relationship, Drug ; Male ; Mice ; Mice, Inbred Strains ; Nickel ; toxicity ; Oxidative Stress ; physiology ; Testis ; drug effects ; metabolism

Objective To assess the effectiveness and safety of embolic protection device in renal angioplasty and stenting.Methods From March 2003 through Feb 2005,renal angioplasty and stenting (RAS)were performed in 73 patients with severe renal artery stenosis,14 of them were done with use of distal embolic protection device(EPD,in 17 arteries ).Technical success included not only the stent placement but also the successful use of EPD.Results The EPD and stents were delivered and deployed successfully in all target arteries.The average percentage of renal artery stenosis before and after stent placement were 80.1%?9.0%,and 6.0%?4.2% respectively.The cholesterol particles were found in the EPD grossly in 2 and microscopically in 9 cases.Conclusion The use of embolic protection device during renal angioplasty and stenting is technically feasible and appears to be effective in preventing procedure-related embolization complications.

Objective To explore the changes of the plasma motilin(MTL)and serum gastrin(GAS)levels and their relationship with course of disease,the severity,the complication of digestive system in neonates with intracranial haemorrhage(ICH).Methods The objects were 26 cases of term newborns with ICH,the healthy control group contained 30 cases of healthy term newborns.Plasma motilin and serum gastrin concentration were measured by radioimmmunoassay in 26 cases of newborns with ICH in the 2 d,3-5 d,7-10 d,and 12-15 d after birth,and compared with the healthy control group.Results Compared with healthy control group,the blood GAS and MTL levels of neonatal ICH group increased dramatically(Pa

Objective To study the characteristics of articles,citations and authors in Chinese Critical Care Medicine (Chin Crit Care Med).Methods The date about articles,citations and authors of Chin Crit Care Med from 2005 to 2006 were collected and analyzed in terms of bibliomertic parameters.Results There were 612 papers published in Chin Crit Care Med during 2 years and the papers with funded projects were accounted for 63.1%.The authors came from 30 provinces,autonomous regions and municipalities directly under the central government in China.The average number of articles per issue was 26;the paper density was 0.40 and the average period from the date of submission to publication was 5.2 months.Among these papers,87.6% were shared with co-authors,with cooperation rate of 4.40.The papers with citation were accounted for 85.1% and 95.9% of the citations in English came from journals.The Price's index was 58.2%.Conclusion The articles published in Chin Crit Care Med have high quality.It is one of the most important resources of information for researches in this field,and it is also a primary kernel journal in Chinese medicine field.