The recombinant chimeric enzyme of AnsB-TTP-P277 comprising L-asparaginase, a tetanus toxin peptide spacer and P277 was expressed as a soluble protein in Escherichia coli. The purified chimeric enzyme exhibited primary activity of the native asparaginase. Prediabetic NOD mice immunized with the chimeric enzyme could induce specific antibodies against P277 and the specificity of anti-P277 antibodies was verified by Western blot assay. The study showed that displaying the P277 epitope on the surface of asparaginase could effectively overcome the weak antigenicity of the P277 epitope and evoke a strong P277-specific immune response in mice. Moreover, the concentration of blood glucose was measured by an automated analyzer.Histochemical analysis of mice pancreas tissues showed that the administration of the chimeric enzyme to NOD mice could prevent the development of diabetes more efficiently than the peptide P277 itself.

A novel fibrinogenolytic protease,named atrase A,has been purified from the venom of Naja atra by sequential chromatography.Atrase A is a single chain glycoprotein with a molecular weight of 64.6 kD,an isoelectric point of pH 9.6 and a neutral sugar content of 4.16%.Atrase A specifically and slowly degraded α-chain of fibrinogen.This fibrinogenolytic activity Was inhibited by chelating agents(EDTA,EGTA and 1,10-phenanthroline)and DTY,and partially inhibited by PMSF,but not by soybean trypsin inhibitor,indicating it is a metalloproteinase.Atrase A showed edema-inducing activity and bactericidal activity against Staphylococcusa aureus.Atrase A did not show cytotoxicity on A549 and K562 cells in MTT assay,but detached adherent A549 cells from the substrate.Atrase A did not show significant inhibition of platelet aggregation induced by ADP or collagen,and did not exhibit proteolytic activities towards fibrin,azocasein and BAEE.It also did not show hemorrhage activity when injected subcutaneously into mice.

Single-nucleotide polymorphisms of the MyoG gene were tested using PCR-SSCP from 73 Landrace pigs, 68 Large White pigs, 57 Yimeng pigs and 83 Laiwu pigs. The effects of the MyoG gene on the birth weight, the average daily gain, the meat tenderness, and the backfat thickness were also analyzed. On the basis of the DNA sequence (M14331) of the porcine MyoG gene, primers were designed to amplify MyoG gene. One polymorphism was found in the amplified region of intron 1, in which two alleles ( A, and B) and three genotypes (AA, AB, and BB ) were examined. A G→ C transition was detected at 2 943 locus by sequencing the homozygotes. The results show that: ( 1 ) The Large White breed differed significantly ( P ＜0.05) in genotype distribution from the Landrace, the Laiwu and the Yimeng breeds;and the Laiwu breed differed significantly( P ＜ 0.05) in genotype distribution from the Landrace and the Yimeng breeds; whereas no significant differences( P ＞ 0.05) were found in genotype distribution between the Landrace and the Yimeng breeds. (2) On the basis of the fixed effect model, significant differences ( P ＜ 0.05) were found in the birth weight and the tenderness among the different MyoG genotypes, whereas no significant differences ( P ＞ 0.05)existed in the average daily gain and the backfat thickness. (3) Using least square analysis, it was seen that significant differences ( P ＜ 0.05) exist in the meat tenderness between the individuals of the BB genotypes and those of the AA genotypes; and significant differences ( P ＜ 0.05 ) were found in the backfat thickness compared the pigs of the AA genotypes with the pigs of the AB, BB genotypes. These results suggest that the MyoG genotype has significant effects on the meat quality, the growth rate, and the backfat thickness,therefore MyoG gene can be used in marker-assisted selection to improve meat quality and growth rate, and to accelerate the breeding progress.

An efficient and repeatable approach in transforming the foxtail millet (Setaria italica ) using Agrobacterium LBA4404 horboring the plasmid pBI121 was established. Factors affecting transformation efficiency were investigated including the genotype, explants, the density of bacteria, the duration for inoculation and co-cultivation, and the concentration of acetosyringone in the medium. The maximum transformation conditions were: the callus induced from inflorescence was used as explant; the duration for inoculation with Agrobacterium at low cell density was 30-40 min; for the co-cultivation , the suitable concentration of acetosyringone in the medium was 0.1 mmol/L, and the duration was 2 days.

Leaf senescence is considered as one of important factors to decrease ornamental values of foliage plants. In the attempt to study and understand the molecular mechanism of leaf senescence, a senescent leaf cDNA library of Coleus blumei was constructed and a small EST library was obtained. According to the sequence of an EST fragment with a cystathionine beta synthase (CBS) domain, a novel leaf senescenceassociated gene (SAG) full-length cDNA encoding a CBS-domain-containing protein, denoted Cbcbs, was rapidly cloned using a strategy of RACE combined with cDNA library. The full length of the Cbcbs gene was 859 bp long (accession No. EF076754) and contained a 609 bp open reading frame (ORF) encoding a 202amino acid protein. One stop codon (TAA) was found in 5' UTR and one possible polyadenylation signal,AATAAA, and a pentanucleotide motif, ATTTA, were found in 3' UTR. The CbCBS contained a predicted mitochondrial targeting peptide in the N-terminal region, two conserved and intact CBS domains, four casein kinase Ⅱ (CK Ⅱ) phosphorylation sites, three protein kinase c (PKC) phosphorylation sites and one tyrosine sulfation (TS) site. Sequence comparisons and phylogenetic analysis showed that CbCBS was a novel senescence or stress-associated protein. The prediction analysis of secondary structure and three dimensional structure of CbCBS suggested that the chief function of the protein was decided by the CBS domain pair. The expression pattern of Cbcbs in leaves was analyzed by RT-PCR. It was demonstrated that Cbcbs gene was a senescence-associated gene (SAG) and expressed in all leaf stages, young stage (Y) being the lowest and terminal senescence stage (S3) being the highest, and was upregulated along with the leaf senescence.Function analysis showed that the mature CbCBS maybe acts as a sensor of cellular energy status and directly or indirectly regulates cellular energy levels to increase ATP content in mitochondria during periods of metabolic stress of senescent leaves.

The kiwifruit germplasms in Jiangxi province were identified by amplified fragment length polymorphism(AFLP)markers．Four primer pairs that had been selected from 64 ones had detected a total of 190 bands among 31 germplasms of kiwifruit,one hundred and seventy nine bands that representing 94．2％of total bands were polymorphic．The identification rates of 31 germplasms were up to 100％．The result suggests that applying AFLP markers to analyze the kiwifruit germplasms is feasible．Then Clustering analysis the AFLP resuit by UPGMA．The dendrogram indicated that the Dice similarity coefficients of 31 germplasms of kiwifruit ranged from 0．50 to 0．85,it suggested that their genetic relationships weren't near．Thirty one germplasms could divide into four groups according to the Dice similarity coefficient 0．56．Sect．Leiocarpae and Sect．Maculatae clustered into one group；A．Melliana of Sect．Strigosae was one group；A．Chinensis var. Rufopulpa,A．Eriantha,A．Fulvicoma var．Lenata,A．Styracifolia and A．Jiangxiensis in Sect．Stellatae clustered into the same group；but the two germplasms of Sect．Stellatae,A．Deliciosa and A．Chinensis clustered into the other group．Interestingly,the‘Ganmi-5’was in the same rank with A．Chinensis from the dendrogram,but it was the variety of A．Chinensis according to the traditional classification．It is necessary that the‘Ganmi-5’should be further researched in classification．The genetic relationships among the kiwifruit germplasms in Jiangxi province were identified and characterized by themolecular method,which was consistent with the traditional classification in certain degree and provided new evidences for the classification of the kiwifruit germplasms．

The effect of recombinant human phospholipase D2(rhPLD2)in vivo was investigated on the secretion of serum glycosyl phosphatidylinositol-specific phospholipase D(GPI-PLD)in guinea pigs of chronic asthma．Ater treating the guinea pigs attacked by chronic asthma with rhPLD2,the GPI-PLD activity detection was canrried out by phase separation of human placental alkaline phosphatase in Triton X-114．Compared with the healthy guinea pigs(NS group),the serum GPI-PLD in the guinea pigs of chronic asthma are much higher than that of control groups,P≤0．01．Our results showed that rhPLD2 could significantly reduce the secretion of GPI-PLD when the guinea pigs were attacked by chronic asthma．

Neprilysin(NEP)is a type Ⅱ integral membrane glycoprotein of the M13 zinc metalloprotease family．As a neuropeptide degrading enzyme,NEP has been discovered to possess an increasing amount of organ-specific functions from central nervous system to peripheral tissues since it was fimfly identified in 1974．For example,NEP has been shown to have an anti-tumour effect by inhibition of cell migration and proliferation while induction of an programmed cell death dependent of both its enzymatic activity and direct protein-protein interactions with key molecules involved in signal transduction pathways:NEP Was also implicated to have neuropmtective effect by preventing the accumulation of the neurotoxic amyloid β-peptide (Aβ)in brain．Through investigating the progression of various human diseases,impaired NEP expression and activity were found to occur frequently．Based on these findings,modulation of NEP levels in pathological cells is considered to be therapeutically applicable as a strategy to recover normal cell functions and thereafter relieve symptoms of diseases．Great research effort is being contributed to the study of regulatory mechanisms involved in expression and activity of this enzyme,and a number of encouraging results have already been achieved．Besides androgens,well-recognised regulators of NEP transcription in prostate,the female hormone oestrogen,aqueous extract of willow herb,components of green tea and neuropeptides bombesin,somatostatin as well as the intracellular domain of amyloid precursor protein were all shown to have a stimulatory effect on NEP expression and／or its activity．

Cancer metastasis is composed of a complex cascade that involves a variety of critical steps beginning with detachment from the primary tumor and ending with growth of tumor at a distant site, such as bone. The "seed-and-soil hypothesis" predicts that the bone microenvironment expresses factors through which attract a variety of cancer cells and promote the tumor development. The ending point of tumor development in bone is achieved through the bidirectional and dynamic interaction between tumor cells and the cells in their growth microenvironment. A variety of factors produced by the bone microenvironment, contribute to the pathogenesis of cancer skeletal metastasis. In this review, using prostate cancer (CaP) as an example, some of general mechanisms of cancer metastasis will be summarized. In addition, the current understanding of the interaction between tumor cells and the bone microenvironment will be addressed. Finally, the research directions in the near future will be suggested.

Several M-CAT elements are present in the chicken nicotinic acetylcholine receptor γ-subunit promoter and required for the totipotency of this gene.To understand the binding properties of M-CAT element,the interaction of M-CAT motif,at the position -67/-61 relative to the initiation site of γ-promoter with nuclear proteins,were analyzed.Mutageneses and binding experiments demonstrated that the M-CAT core motif bound nuclear protein (s) from tested chicken tissues to generate a high-mobility complex;while the flanking sequence bound different factors to generate lower-mobility complexes with the exception of liver.Southwestern blotting showed that only a 30 kD protein was expressed in tested tissues of chicken embryo at day 13.These results suggest that a 30 kD factor can directly bind to M-CAT element in the manner of monomer,even homodimer and is ubiquitously expressed in tissues,while the binding of the factor(s) to flanking sequence should probably rely on the presence of a 30 kD ubiquitous factor.