To investigate the profile of gene expression in Salvia miltiorrhiza and elucidate its functional gene, 454 GS FLX platform and Titanium regent were used to produce a substantial expressed sequence tags (ESTs) dataset from the root of S. miltiorrhiza. A total of 46 722 ESTs with an average read length of 414 bp were generated. 454 ESTs were combined with the S. miltiorrhiza ESTs from GenBank. These ESTs were assembled into 18 235 unigenes. Of these unigenes, 454 sequencing identified 13 980 novel unigenes. 73% of these unigenes (13 308) were annotated using BLAST searches (E-value < or = 1e-5) against the SwissProt, KEGG TAIR, Nr and Nt databases. Twenty-seven unigenes (encoding 15 enzymes) were found to be involved in tanshinones biosynthesis, and 29 unigenes (encoding 11 enzymes) involved in phenolic acids biosynthesis. Seventy putative genes were found to encode cytochromes P450 and 577 putative transcription factor genes. Data presented in this study will constitute an important resource for the scientific community that is interested in the molecular genetics and functional genomics of S. miltiorrhiza.

Synthetic biology of traditional Chinese medicine (TCM) is a new and developing subject based on the research of secondary metabolite biosynthesis for nature products. The early development of synthetic biology focused on the screening and modification of parts or devices, and establishment of standardized device libraries. Panax notoginseng (Burk.) F.H.Chen is one of the most famous medicinal plants in Panax species. Triterpene saponins have important pharmacological activities in P. notoginseng. Squalene epoxidase (SE) has been considered as a key rate-limiting enzyme in biosynthetic pathways of triterpene saponins and phytosterols. SE acts as one of necessary devices for biosynthesis of triterpene saponins and phytosterols in vitro via synthetic biology approach. Here we cloned two genes encoding squalene epoxidase (PnSE1 and PnSE2) and analyzed the predict amino acid sequences by bioinformatic analysis. Further, we detected the gene expression profiling in different organs and the expression level of SEs in leaves elicited by methyl jasmonate (MeJA) treatment in 4-year-old P notoginseng using real-time quantitative PCR (real-time PCR). The study will provide a foundation for discovery and modification of devices in previous research by TCM synthetic biology. PnSE1 and PnSE2 encoded predicted proteins of 537 and 545 amino acids, respectively. Two amino acid sequences predicted from PnSEs shared strong similarity (79%), but were highly divergent in N-terminal regions (the first 70 amino acids). The genes expression profiling detected by real-time PCR, PnSE1 mRNA abundantly accumulated in all organs, especially in flower. PnSE2 was only weakly expressed and preferentially in flower. MeJA treatment enhanced the accumulation of PnSEI mRNA expression level in leaves, while there is no obvious enhancement of PnSE2 in same condition. Results indicated that the gene expressions of PnSE1 and PnSE2 were differently transcribed in four organs, and two PnSEs differently responded to MeJA stimuli. It was strongly suggested that PnSEs play different roles in secondary metabolite biosynthesis in P. notoginseng. PnSE1 might be involved in triterpenoid biosynthesis and PnSE2 might be involved in phytosterol biosynthesis.

ERF family transcription factor (TF) represented ethylene-responsive protein which harbored a conserved AP2 domain. After searching the plant transcription factor database, a total of 75 unigenes was found which contained AP2 domain from the transcriptome dataset of Panax quinquefolius L. One unique sequence of ERF transcript, named as PqERF1, was cloned with entire open reading frame of 933 base pairs (bp). Protein prediction result indicated that the gene was localized in nucleus and had a conserved AP2 domain. PqERF1 gene could be induced by methyl jasmonate (MeJA) which was consistent to the inducing profile of triterpene ginsenosides. InterproScan prediction indicated that PqERF1 was probably a pathogenesis-related gene. Sequence alignment and phylogenetic analysis demonstrated PqERF1 was with high identity and had relative close relationship to the NtERF4 (Nicotiana tabacum), PhERF12 (Petunia x hybrida) and DcERF1 (Daucus carota) which was related to plant defense, regulation of secondary metabolism and the flower senescence respectively. Therefore, the gene was likely involved in regulation of secondary metabolism, plant defense and physical processes which would provide gene resource for further study on secondary metabolite synthesis and molecular breeding of P. quinquefolius.

ObjectiveTo study the relationship between the antitumor immunoreaction and the cell apoptosis in esophageal carcinoma.MethodsThe IL-1β converting enzyme, Fas and FasL protein were labeled by LSAB immunohistochemistric method in 46 cases of esophageal carcinoma, of which 13 cases were labeled by TdT mediated dUTP nick end labeling (TUNEL).ResultsThe positive rates of ICE, Fas and FasL proteins in the cancer nests were 78.26%, 60.87% and 47.83%. The expression of ICE protein was related to the histological grading of the cancer.Conclusions The expression of Fas, FasL protein may be related to the immunological escape of the cancer cell; the expression of ICE was related to the histological grading of the cancer.

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.

Objective: To explore the effects of individual materialism, social status on victimization, and to provide a reference for the intervention of campus bullying. Methods: A total of 2 597 grade 7 students representing 47 classes from 7 junior middle schools in Zhenzhou were administered with Material Values Scale, Peer Nomination Questionnaire and Victim Questionnaire. Hierarchical Linear Model was used in data analysis. Results: Peer rejection(γ=0.15, P<0.01) and individual materialism(γ=0.13, P<0.01) positively predicted victimization, while popularity negatively predicted victimization(γ=-0.05, P<0.01). Class materialism norm also could positively predict victimization(γ=0.82, P<0.01). Moreover, there was a significant interaction between class materialism norm and peer rejection(γ=0.30, P<0.05), and the results of simple effect showed that with the increase of materialism level, the negative impact of peer rejection on victimization was increasing(γ high=0.18, Z high=7.80; γ low=0.12, Z low=5.50, P<0.01). Conclusion Peer rejection, individual materialism, popularity and class materialism norm affect individual bullying, and class materialism norm could moderate the relationship between peer rejection and victimization.

OBJECTIVETo clone and sequence the open reading frame of cycloartenol synthase (CAS) from Huperzia carinata.

METHODAfter searching the transcriptome dataset of H. carinata, one unique sequence containing oxide squalene cyclases domain was discovered. The primers were designed according to the cDNA sequence of CAS from the dataset. And then, the open reading frame of CAS was cloned by RT-PCR strategy with the template of mixed RNA extracted from root, stem and leaf of H. carinata. The bioinformatic analysis of this gene and its corresponding protein was performed.

RESULTOne unique sequence of CAS, named as HcCAS1 (GenBank accession number JN790125) , was cloned from H. carinata. The open reading frame of HcCAS1 consists of 2 474 bp, encoding one polypeptide with 757 amino acids.

CONCLUSIONThis study cloned and analyzed CAS from H. carinata for the first time. The result will provide a foundation for exploring the mechanism of sterol biosynthesis in Huperziaceae plants.

Amino Acid Sequence ; Cloning, Molecular ; Computational Biology ; Evolution, Molecular ; Huperzia ; enzymology ; genetics ; Intramolecular Transferases ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Protein Structure, Secondary