A serum inhibited gene Si1(GenBank acession number:AY050169) was previously cloned and identified by differential expression of genes in U251 cells.For the further study of biological function of Si1, prediction procedure was performed to predict its subcelluar-localization.Relative experiments were carried out at the same time.The expression of EGFP/Si1 recombinant in HeLa cells showed Si1 protein located in nuclear which corroborated the prediction results of PsortⅡ, Proloc, Cello version2, Subnuclear compartments prediction system, NUCLEO and NUCPRED.According to the PredictNLS prediction, twelve different fragments of EGFP/Si1 recombinants were constructed to identify precise NLS regulation sequence.Findings proved that the real NLS regulation sequence was not the same as the software predicted(1 206 bp～1 239 bp on Si1 ORF), but located on 1 395 bp～1 594 bp of Si1.A tumor relatived mutation/EGFP recombinant localization result showed though the mutation site(1 639bp on Si1 ORF) does not located in NLS regulation sequence, it did affect wildtype Si1 protein divert to nuclear and may affect its natural function in cell, perhaps it is the main reason for highly mutation rate of Si1 in tumor.

Embryonic stem cells(ESCs) are pluripotent cells derived from the inner cell mass of blastocyst-stage embryos, possessing permanent self-renewal and indefinite proliferative capacity in vitro. And ESCs could differentiate into the hematopoietic cell fate. Therefore ESCs may provide an alternative for hematopoietic stem cell transplantation and blood cells transfusion. Furthermore, ESCs differentiation could also provide a powerful model system to better understand the hematopoietic development and the mechanism involved. The current status for efforts to differentiate ESCs into hematopoietic lineages were reviewed.

The modern large-scale pyrosequencing technology is a revolution of DNA sequencing.One of the key points in this technology is to get an ATP sulfurylase immobilized on the surface of magnetic beads and with a high activity.Biotinylated ATP sulfurylase can be immobilized on magnetic beads coated with streptavidin through the specific conjunction between biotin and streptavidin, but using chemical modification method to biotinylate ATPS will affect the activity of the enzyme.ATP sulfurylase fused with the carboxyl terminal 87 residues of Escherichia coli biotin carboxyl carrier protein(BCCP87) was expressed in E.coli using fusion expression strategy.Results from Western blot analysis and SDS-PAGE analysis showed that the fusion protein could be biotinylated in vivo, and the molecular mass of the fusion protein was about 64 ku.The biotinylated ATP sulfurylase could be immobilized on the surface of magnetic beads coated with strepavidin, and the immobilized ATPS could be used for quantification of PPi and pyrosequencing.An effective enzyme for the large-scale chip-based pyrosequencing system was supplied.

Baculovirus-mediated gene transfer into mammalian cells has been used to develop non-replicative vector vaccines against a number of diseases in several animal models.A baculovirus pseudotyped with the glycoprotein of vesicular stomatitis virus was used as vector to construct the recombinant baculovirus expressing classical swine fever virus(CSFV) E2 protein under the control of ie1 promoter from white spot syndrome virus.The E2 gene was shown to be efficiently expressed in both insect and mammalian cells.Intramuscular injection of mice with the recombinant baculovirus resulted in the production of high-level CSFV-specific antibodies.Specific lymphoproliferative responses to the CSFV stimulation were induced in the splenocytes of the immunized mice as demonstrated by CFSE staining assay and WST-8 assay.The results indicates that the pseudotyped baculovirus-delivered gene can be a potential non-replicative vaccine against CSFV infection.

Analysis of regular elements in promoter region is the base for elucidating the mechanism of gene transcription initiation.The TATA and the TATA-less promoters of plant RNA polymerase Ⅱ gene are chosen from the PlanPromDB.The GC bias,position structure conservation,nucleotide content and conservative motifs of sequences,position distribution of TATA box and conservation of correlation position are analyzed.Many specific regulars for the two types of promoters are found.These features can offer some help for revealing the transcription regulation of plant gene.A new prediction algorithm based on position-correlation weight matrix(PCWM) is proposed.The better discrimination results for two sort plant promoters are obtained by using score function.It is confirmed that the performance of position-correlation weight matrix(PCWM) is superior to single-base position weight matrix(PWM).

The process of protein synthesis is terminated by one of the three stop codons which are recognized by classⅠ polypeptide release factors.Subsequently, it could promote the hydrolysis of the ester bond of peptidy-tRNA, resulting in release of the nascent polypeptide.Recent results from cryoelectron microscopy, crystallography, NMR, molecular dynamic and biochemical experiments have shed considerable light on the function and structure of the classⅠ release factors.The progress in these aspects were summarized.

Echinoderms are deuterostome, and occupy the top taxonomic position of invertebrates, where is the evolutionary linkage between invertebrates and vertebrates.This is a critical phylogenetic vantage point to infer both the early evolution of bilaterian immunity and the underpinnings of the vertebrate adaptive immune system.The available published literatures on echinoderm immunological mechanisms are compiled and discussed here to understand its evolution process and the hot spots or directions in future investigations.Echinoderms have an innate immune system like vertebrates but their adaptive attributes have not been observed.Its immune responses are based on coelomocytes activity working in parallel with a variety of humoral factors that react directly with invading pathogens.Comparative studies of immune functions in echinoderms have demonstrated that there is a complement system in echinoderms that appears to have alternative pathway and lectin pathway but lack classical pathway and terminal pathway.The sea urchin innate immune system has a number of large gene families.Further investigations are needed to understand the unknown immune-associated genes, proteins, immune signaling pathways and effector molecules, and to know the origin, the underlying mechanisms and evolution of innate immune system.

Previous studies suggest that NGAL (neutro phil gelatinase-associated lipocalin) is involved in the transformation and development of esophageal carcinoma. Alteration of NGAL expression can trigger the change of cellular morphology in esophageal carcinoma cells. However, the mechanisms remain unclear. To get a better understanding of NGAL function in esophageal carcinoma, NGAL protein was expressed in methylotrophic yeast, Pichia pastoris, and purified by chromatography. EC1.71 cells expressed high levels of NGALR (NGAL receptor) and EC109 cells expressed low levels of NGALR were used as cells model. The trafficking and the possible function of NGAL protein were then analyzed in the esophageal carcinoma cells. The results showed that 5-FAM-labeled recombinant NGAL protein could internalize into the EC1.71 and EC109 cells. Furthermore, the internalized NGAL protein could induce the alteration of cellular morphology, resulting in generation of autophagosome, transcriptional up-regulation of genes associated with autophagy and increase of phospho-ERK1/2 (p-ERK1/2). Interestingly, the treatment with the NGAL protein did not affect the intracellular iron level. These data indicate that induced autophagy by exogenous NGAL protein is a mechanism that internalized NGAL plays important roles in esophageal carcinoma cells, independent with NGAL-mediated iron transport process, while ERK1/2 signal pathway is involved in activation of autophagy by exogenous NGAL protein.

In order to investigate if there exists interaction between mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3) protein, and how the interaction regulates tumor necrosis factor-? (TNF-?) transcription activity, the human p38 and extracellular-signal regulated protein kinase 2 (ERK2) genes were amplified from human flag-p38 and flag-ERK2 by polymerase chain reaction (PCR) and cloned into pcDNA3-HA. Protein expression of the plasmids was examined by Western blotting. Co-immunoprecipitation was used to identify if there exists interaction between MAPK and STAT3 proteins. If the interaction was approved to be true, report gene system was applied to find how the interaction affect transcriptional expression of TNF-?. After STAT3 pathway was inhibited by RNA interfering, the action on TNF-? activity was determined. The results of DNA sequencing and enzyme digestion showed that the cloned p38 and ERK2 genes were correct, to be 1 080 bp or so. p38 and ERK2 proteins were expressed in 293T cell to be approximately 40 ku. Co-immunoprecipitation data showed that p38 and ERK2 proteins integrated with STAT3 protein in vivo. TNF-? reporter gene activity results found that protein complex of p38-STAT3 and ERK2-STAT3 coordinately increased TNF-? activity. After STAT3 was interfered, the TNF-? activity markedly decreased. These data indicated that there exists interaction between p38 and STAT3 protein, ERK2 and STAT3 protein. The complex of the proteins can coordinately regulate TNF-? expression. After interfereing STAT3 pathway, the coordinated action on TNF-? transcription activity might be obviously reduced.

The pandemic outbreak of influenza has been started from Mexico in 2009 to 70 countries during 2 months. On 11th of June , WHO announced influenza pandemic alert level rose to the highest level 6, which means the first influenza pandemic in 21st century is coming. Till 6th of July, 94 512 confirmed cases from more than 120 countries and areas were reported, including 429 cases were died. The genetic fragment of swine, poultry sources and human influenza viruses are contained in this strain, A/H1N1 influenza virus, of the pandemic. It is of great significance of studying the genetic reassortment, evolution and its biological characteristics of this virus strain to prevent and control the pandemic. At present, the genetic evolution of strain has been identified, and the potential biological characteristics have been analyzed by genetic traits, however, clinical manifestation should be further concerned, and the tendency of influenza pandemic and genetic changes need to be monitored closely. The complexity of influenza virus ecosystems, mutation of genome, and easy to preserve in "Nature Gene Pool" and reassortment, make the influenza pandemic inevitable. We should face the threat of influenza pandemic, enhance the surveillance of influenza virus in ecosystems, strengthen the epidemiological investigation, develop the vaccines and drugs, and establish an effective public health security system, in order to reduce the destruction of the influenza pandemic.