Abstract:The completion of sequencing human genome creates a new era of biological science and technology. Although the sequence of the human genome has been known, it is still hard to rapidly explore the whole functional genes, especially, their interaction with each other and the meaning to the body. However, the “reverse biology” which comes into being in the recent years provides us a series of novel ideas and technologies for discovering new functional gene, among which the immune-related genes have attracted more attentions, clarifying how functional gene works and their potential value in application.

Abstract:Histone methylation is one of the epigenetic modifications. Histone methylation influences constitutive heterochromatin, genomic imprinting, inactivation of X-chromosome and gene transcription regulation. Abnormality of histone methylation is associated with several carcinomas. The discovery of enzymes that reverse histone methylation challenges the current understanding that histone methylation is a stable epigenetic marker and provides a novel way to study histone modifications.

Abstract:Recently the research of vaccines and drugs against anthrax is one of hot spots. The efficacy of anthrax vaccines and drugs can't be experimented in human, therefore the testing model is very important. The cell models mainly include CHO and J774A.1. Now, various kinds of animals including mice, rats, rabbits, and nonhuman primates were experimented as animal models. Because the models are different, the results of experiments are significantly different, sometimes they are contrary. Many experiments of Bacillus anthracis in different cell and animal models are reviewed, and the principles of choosing animal models of anthrax are discussed. In order to analyze the different results of experiments in different models, the pathogenesis of Bacillus anthracis and the researching progress of anthrax vaccines and drugs are also simply introduced.

Abstract:Trophoblast invasion during implantation mimics the process of tumor metastasis in some aspects, however with a significant difference in that the former is strictly controlled. Several members of matrix metalloproteinases (MMPs) family have been shown to play crucial roles in regulating trophoblast cell invasion. MMP-26 is a newly identified MMP family member, and its functions in human trophoblast cells have yet to be elucidated. A well-accepted human trophoblast cell model———choriocarcinoma cell line (JEG-3) was used as in vitro model to investigate the involvement of MMP-26 in trophoblast cell invasion. Expressing pCR3.1 plasmid containing MMP-26 cDNA was transfected into JEG-3 cells, and a stable cell line with over-expressed MMP-26 was named as JEG-3/MMP-26. Transwell insert invasion assay showed that cell invasiveness in JEG-3/MMP-26 cells was significantly promoted. Data of RT-PCR and gelatin zymography demonstrated that the mRNA expression and zymogen secretion of MMP-9 were increased in these cells. Double immunofluorescent staining revealed co-localization of MMP-26 and MMP-9 proteins in JEG-3/MMP-26 cells. The data indicated that MMP-26 was involved in invasion-promoting regulation of human trophoblast cells, and it may function, at least in part, through coordination with other MMPs such as MMP-9.

Abstract:BRD7, a novel bromodomain gene, has been cloned by cDNA RDA (cDNA Representational Difference Analysis). The GenBank accession number is AF152604. Previous studies showed that BRD7 gene was down-regulated in nasopharyngeal carcinoma (NPC) cells and tissues. Over expression of BRD7 in NPC cells can inhibit cell proliferation and cell cycle progression from G1 to S phase, and can partly inhibit the aberrant growth of NPC cells. In order to explore the molecular mechanisms that involved in the down-regulation of BRD7 gene in NPC cells, bioinformatic approaches were adopted and a putative promoter region was found. The luciferase expression vectors containing BRD7 promoter region were further constructed . Transient transfection results suggested that the analyzed sequence contained BRD7 promoter. Transcriptional factor Sp1 is responsive to this region. Inhibition of the Sp1 binding to BRD7 promoter by mithramycin A significantly reduced the promoter activity and the endogenous expression of BRD7 in mRNA level.

Abstract:In order to examine potential molecular markers of pancreatic stem cells, a regeneration pancreatic model was induced by 90% partial pancreatectomy (Px) in rats. Changes in the protein expression in rat regeneration pancreas at 3rd day after Px, as comparing to sham surgery (Sx), were analyzed by using two-dimensional gel electrophoresis (2DE), mass spectrometry (MS), and peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database searching. HSP47 and Vimentin were analyzed in Px and Sx rat with Western blotting to confirm the results of 2DE. The average spots in gels for Sx and Px pancreatic tissues were (1 369 ± 31) and (1 315 ± 28), respectively. 2DE displayed 91 spots with at least 1.5-fold of differential expression at the time point of 3 days after pancreatectomy and 53 differentially expressed proteins were identified by PMF. Among them, Vimentin, CK8, L-plastin, hnRNP A2/B1 and AGAT are associated with embryogenesis and cells differentiation and may be new potential pancreatic stem cells markers. The proteome profiling technique provided a broad-based and effective approach for discovering potential biomarkers of pancreatic stem cells.

Abstract:The defects and dysfunctions in antigen-presenting dendritic cells have been shown during carcinogenesis leading to human immune system can be markedly suppressed by cancer. To explore the mechanisms by which cancers escape from immune recognition, the influences of tumor microenvironment on differentiation of dendritic cells (DCs), which play an important role in tumor immunology, were studied by biophysical and microrheological methods. It was found that the components derived from tumors caused the increases in osmotic fragility and decreases in membrane fluidity of DCs, moreover, the gene transcriptional levels and energy statuses of cells were also transformed, leading to impaired capabilities of antigen uptaking and deteriorated stimulatory capacities to activate na?觙ve T cells. The impaired microrheological properties of DCs may be one of the aspects of immune escape mechanism of tumor.

Abstract:In multiarray experiments, there is some system bias, which contaminated by experimental factors such as spot location (often referred to as a print-tip effect), arrays, dyes, and various interactions of these effects. For comparable each other, it is necessary to normalize the raw expression profile data. Normalization is the key step in low level processing. In fact, many normalization methods have been developed, i.e. Scaling normalization, Nonlinear normalization, Quantile normalization and so on. New baseline normalization is presented. First, select the subset of probes, which have the min rank range. Second, do nonlinear normalization on robust baseline. Iterative strategy weakens the sensitivity of the baseline method to select baseline. With the standard test dataset, compare it with other methods. The results show that the novel method has better performances than others in several ways.

Abstract:Screen small molecule peptide specific binding to small cell lung cancer cell (DMS53) was screened by using the “one-bead one-peptide” combinatorial technology. Thirty two positive beads binding to DMS53 were totally obtained after primary screening. Consensus peptide sequences of cXNGRXXc and cNGRXXXc were identified by amino acid sequencing in ten beads. Three representative peptides were re-synthesized on beads. Secondary screening showed that cell adhesion percentage of cFNGRQQc to DMSS was higher than the other two peptides. cFNGRQQc was further studied for cell specificity, alanine scanning and site-directed deletion. The results showed that cFNGRQQc is specific for promoting cell adhesion to DMS53 but not to other human cell lines. Both motif of -NGR- and the length of six peptide of cFNGRQQc structure are important for DMS53 attachment. In an antibody or peptide blocking assay, cell adhesion of DMS53 to peptide bead was not inhibited by antibodies or peptides including anti-integrin, E-cadherin, NCAM and ICAM. The binding site on DMS53 surface for cFNGRQQc peptide need to be proven in the future.

Abstract:The mechanism of complex unfolding process of Pseudomonas aeruginosa apoazurin is an arguing problem. Recent published results indicated that this problem could be resolved by hypothesizing two native conformations coexisting in solution. Urea-induced unfolding of apoazurin was investigated further using intrinsinc fluorescence and CD spectra. Equilibrium unfolding curves in urea could be depicted with an apparent two-state transition, but a biphase kinetic process. The fast unfolding process is finished within a few seconds as monitored by stopped-flow fluorescence intensity, whereas the slow process requires several hours for unfolding at high concentration of urea at room temperature. The m_u values for the fast and slow phases are 2.24 and 2.45 kJ·mol^－1·M^－1 respectively. The difference between their unfolding activation energy is 22 kJ·mol^－1. The time-resolved fluorescence emission spectra as well as the circular dichroism spectra just after manual mixing of protein and denaturant could be simulated by superposition of the spectra of the native and fully unfolded protein with the same coefficient of 0.37. This strongly suggests the three-state mechanism with a partially unfolded intermediate on its pathway is inadequate. The hypothesis of two native conformations coexistence is a reasonable selection.

Abstract:According to the results of quantum chemistry calculation and the present research status in the relationship between the structures and the functions of DT, the E154 in DT catalyzing domain was mutated to aspartic acid and arginine in order to study the effects of the alteration on the biological activities. By means of gene site-direct mutation, two mutated genes were prepared and the high performance expression was obtained in E.coli system. The results of toxcity studies indicated that the acute toxicity in guinea pig and cytotoxicities of mutant E154D increased slightly in compared with those of recombination wild toxin, and contrarily, those of E154R decreased obviously.

Abstract:A novel method for quickly cloning genes with multiple DNA fragments———one step cloning technique using isoschizomer-heterotail restriction endonuclease (IHRE) is described. Up to six DNA segments are ligated by using only one restriction endonuclease in this method. Comparing with routine method，it is simple, fast, economical and generates products with higher purity and achievement. Light chain of human enterokinase, DNA multi-epitope vaccine to HSV2 have been designed and successfully constructed via this method.

Abstract:Bimolecular fluorescence complementation (BiFC) assay is an innovative approach to investigate protein interactions, based on the reassembly of protein fragments of the fluorescent proteins which directly report interactions. The fluorescent proteins tolerate circular permutation and insertions of foreign proteins with maintenance of fluorescence. So when the proteins fused to the reporter fragments interact with each other, a direct readout of the association would be given from the facilitating reassembly of the active reporter protein. Moreover, with distinct spectra difference of the fluorescent protein family members, BiFC assay is expanded to multicolor BiFC assay which enables visualization of interactions between different proteins in the same cell and comparison of the efficiencies of complex formation with alternative interaction partners. From it first reported in Molecular Cell in 2002 till now, this approach has been used on the networks of protein interaction in mammal cells, plant cells or even E.coli, and researches on transcription factors, G protein βγ dimmers, protein ubiqutination and so on.

Abstract:With the Science Citation Index Expanded（SCIE）and Journal Citation Reports(JCR), the amount and distribution of the originals in Progress in Biochemistry and Biophysics, which were cited by the journals included by SCIE, were analyzed according to method of citation analysis. The average cited number of original articles cited by other researchers is 1.70. The cited authors are from 23 provinces and from other countries. Beijing, Hunan, and Guangdong province are in the front of most cited author's areas. The most frequently cited institute was Xiangya School of Medicine at Central South University. There are 115 citing journals.