Toll-like receptor 4 (TLR4) is mainly expressed on microglia in the central nervous system initiating the innate immune response in mammals. Recently, studies showed that TLR4 is involved in the pathogenesis of pain and inflammation. Morphine can activate TLR4. This results in the activation of microglia and increased synthesis and release of inflammatory cytokines, leading to the hyperexcitability of nociceptive neurons and reduction or abolishment of morphine-induced analgesia, i.e. morphine tolerance. Blockade of TLR4 can enhance the morphine analgesia and inhibit morphine tolerance. TLR4 differs from classical opioid receptors in that TLR4 does not display specific stereoselectivity for opioid ligands. Both (+)-and (-)-morphine can activate TLR4 signaling. The study of interaction of morphine-TLR4-glia may provide a new therapy to inhibit morphine tolerance.

Persistent infection with high-risk types of human papillomavirus has been proved as an important etiological factor for cervical cancer. HPV major capsid protein (L1) can self-assemble into VLPs when expressed in a recombinant expression system. VLP can induce high levels of typespecific neutralizing antibodies to efficiently prevent the infection of the same genotype HPV and eventually prevent cervical cancer and other infection related diseases. HPV type 58 is one of the most common high-risk HPVs in Chinese cervical cancer patients. However, there is no effective vaccine to prevent HPV 58 infection so far. The HPV 58L1wt gene was modified by several strategies. The modified gene, named as HPV 58 mL1, was expressed by Baculovirus Expression System, and the recombinant protein was purified by CsCl ultracentrifugation. The transmission electron microscopy analysis showed that the recombinant HPV 58 mL1 protein could selfassemble into regular VLP with a diameter of about 55 nm. New Zealand white rabbit and guinea pigs were subcutaneously immunized with HPV 58 mL1 VLP. VLP ELISA showed that HPV 58 VLP could induce high titers of HPV 58 mL1 VLP specific antibodies. Dot blot assay indicated that the sera antibodies were against the surface epitopes of VLP. Taken together, HPV 58 mL1 VLP was produced by Baculovirus Expression System and antisera were obtained from two different species, which were important for further developing effective HPV 58 VLP based prophylactic vaccines.

Phosphoinositide 3-kinases (PI3Ks) is one of the main downstream molecules of receptor tyrosine kinases and G protein-coupled receptors. Activated PI3Ks catalyzes the production of second messenger phosphatidylinositol-3,4,5-trisphosphate (PIP3), which phosphorylates Akt. Activation of Akt plays vital roles in regulation of cell proliferation, differentiation, apoptosis, glucose transportation and metabolism via its downstream enzymes or transcriptors such as glycogen synthase kinase-3, FoxO1, and mammalian target of rapamycin (mTOR). The current review aims to briefly summarize and discuss the recent findings regarding the roles of PI3K-Akt signaling axis in regulation of glucose homeostasis.

The Gene Expression Omnibus (GEO) database, the first public repository for gene expression data, premiered at National Center for Biotechnology Information（NCBI） in July 2000. The GEO database contains a wide assortment of high-throughput experimental data, including single and dual channel microarray-based experiments measuring the abundance of mRNA, genomic DNA and protein molecules. Data are also archived which origin from non-array-based high-throughput functional genomics and proteomics technologies, including serial analysis of gene expression (SAGE) and protein identification technology. To date, the GEO database contains data representing almost 10 000 hybridization experiments and SAGE libraries from 30 different organisms. This paper outlines the query and browse in GEO database, data download, format, data analysis, and deposit and update. Also, it focuses on the managing terminology used in the GEO-data browser, whiledescribing the course of data mining and GEO’s future applications in the fieldof molecular biology. GEO is publicly accessible at http:// www.ncbi.nlm.nih.go v/projects/geo/.

The nuclear pore complex (NPC) is in charge of the exchange of macromolecules between the nuclei and the cytoplasm of eucaryotic cells. The small compounds can get into the nuclei through the NPC freely or by passive diffusion. However, those proteins whose molecular weights are larger than 50 kD can only get into nuclei by active transport. There must be one or more special signals in the amino acid sequences, so-called nuclear localization signals, which can be recognized by respective karyopherins, in those proteins. There are diverse sequences in NLSs, including classical NLS (cNLS), PY-NLS, which can be recognized by importin β2, and some other NLSs. Though there are similar properties among the same family members of NLSs, there is no fully conserved amino acid sequence. Different NLSs often correspond to different mechanisms of nuclear transport, andalso, couple of functional NLSs may exist in one certain protein. The studies on NLS help us not only reveal new nuclear transport mechanism of macromolecules, but also discover new functions of proteins. Categories of NLSs were summarized by the authors. Two common-used software employed in NLS prediction and the strategies used to identify NLS are also introduced in this review.

Autophagy is a very important biological process, by which the cytoplasmic constituents of protein and organelles are sequestered in a doublemembrane autophagosome and then delivered to the lysosome for degradation. Mitophagy is a process referred to mitochondria degradation via autophagy. The selective degradation of mitochondria is highly regulated by various molecules, including “core” Atg complexes, Atg32, Atg33, Uth1 and Aup1 that localized on the mitochondrial outer membrane in yeast, as well as NIX, PINK1 and Parkin in mammalian cells. As it is an essential cellular pathway to eliminate damaged or depolarized mitochondria to maintain the homeostasis of cells, the dysfunction of mitophagy is closely related to neurodegenerative diseases, such as the Parkinson disease. This review summarizes the latest advances in mitophagy researches.

In the past years, proteomics was developed quickly and its corresponding methodological research has also been made great progress, a series of new technologies blended in proteomics technology, particularly the new combination of multi-dimensional liquid chromatography and mass spectrometry (MS)-based technologies with protein fragmentation methods, enable us easily to identify the dynamics of proteins profiles in any complex biological process. Isobaric tags for relative and absolute quantification (iTRAQ) is one of the new techniques with high sensitivity and accuracy, demonstrating a remarkable advantage in simultaneous analysis of multiple samples and subsequently providing the relative quantification on hundreds of proteins at one time. The iTRAQ reagent produced high quality, reproducible result in enriched complexes, organelles, and whole cell lysates．Hence iTRAQ has been widely used in areas of life science. This review will focus on the principle, experimental procedure and application of iTRAQ in the last few years.

A number of proteins in eucaryon contain zinc finger domains and this class of proteins are called as zinc finger proteins. Because of this special finger-like structure,zinc finger proteins play an important role in the recognition and binding to DNA, protein and RNA. Many zinc finger proteins contain DNA-binding domains included in the zinc finger domains, and also effect domains such as KRAB, SCAN, BTB/POZ, SNAG, SANT and PLAG, linked to the zinc finger domains. These proteins function as transcriptional factors to regulate the target gene transcription. Some zinc finger proteins such as LIM-ZF, MYND-ZF, PHD-ZF and RING-ZF, function as the protein adaptors to mediate the protein-protein interaction. Additionally, some of zinc finger proteins can interact with RNAs and regulate gene expression at the post-transcriptional levels. Here we review the interaction between zinc finger proteins and DNA, RNAs and other proteins, respectively.

The better understanding of protein-protein interactions is crucial for elucidating the structural/functional relations of proteins, investigating their roles in associated disease development, and determining potential drug targets for clinical applications. A vast number of protein-protein interactions have been identified and the information was organized and hosted in many protein-protein interaction databases with the help of high throughput screening technologies, computational predictions and literature-mining processes. This review will first introduce the three common methods used to acquire protein-protein interaction data in large-scale, and then will focus on the introduction of public databases for protein-protein interactions in humans, including HPRD, BIND, IntAct, MINT, DIP, MIPS, as well as the interconnection and the cross-reference of these databases. Finally we will discuss the application of these databases for building protein-protein interaction networks.

Lactoferricin is an multifunctional polypeptide derived from the N-terminal region of intact lactoferrin through pepsin hydrolysis under acidic conditions. In some cases, the activity of lactoferricin is more potent than that of lactoferrin, including the anti-bacterial (gram-positive or negative), antifungal, antiviral, antitumour, immunomodulatory and anti-inflammatory activities. Comparing to the verified effects of lactoferricin by in vitro experiments, its in vivo effects is being investigated, involving genomic and proteomic approaches for insight mechanistic studies. This article reviews the structure, biological activities and mechanisms of lactoferricin,as also projects its prospects in the preparation and applications．

microRNAs（miRNAs） are newly discovered 21～25nt small RNAs that regulate gene expression at post-transcriptional level. miRNAs have been proved to be involved in various physical and pathological precesses, such as development, cell proliferation, apoptosis, fat metabolism, hormone secretion and tumor development. The study methods on miRNAs include two main categories: miRNA-specific technology based on classical experiment techniques and broadly applied bioinformatics. The former is involved in miRNA detection and function analysis, and the latter focus on miRNA gene prediction and miRNA targets prediction. These two complementary parts could provide not only the functional clues, but also the solid proof to better understand the physical roles of miRNAs and the mechanisms by which they function.

Initiation translation of most eukaryotic mRNAs occurs by cap-dependent translation. However, a group of positive sense RNA viruses lacking cap structures can initiate the translation by a cis-element in the 5′untranslated region——internal ribosome entry site (IRES). They can recruit ribosome subunits to the start site of viral mRNA with the aid of certain trans-acting factors. To date，RNA viruses of IRES-dependent translation initiation are discovered in the mammals, invertebrates and plants. IRES elements are grouped in four classes based on the different structure and function. The further study of IRES elements in the RNA viruses will not o nly help the understanding viral pathogenesis, but also provide the guidance for the industrial application and disease therapy. The review focuses on the discovery, classes, structure and function of IRES elements,respectively.

For the industrial enzymatic production of D-p-hydroxyphenylglycine (D-p-HPG), substrate D,L-p-hydroxyphenylhydantoin (D,L-p-HPH) was synthesized using glyoxylic acid-urea-phenol, the enzymatic conversion of D,L-p-HPH to intermediate N-carbamoyl-D-p-hydroxyphenylglycine (CpHPG) was carried out by D-hydantoinase genetic engineering strain E. coli BL21/pMD-T 7-dht resting cells and the chemical conversion of CpHPG to D-p-HPG was performed using sodium nitrite under acidic conditions. The results showed that chemical yield of D, L-p-HPH was 60%. The fermentation was carried out for 12 hours in a 10 L fermentor containing 8 L medium. The D-hydantoinase activity of engineering strain E. coli BL21/pMD-T 7-dht was 3000 U/L and the amount of the recombinant D-hydantoinase was about 60% of the total bacterial soluble proteins by SDS-polyacrylamide gel electrophoresis thin layer scanning. The yield of bacterial cell was 6%. Biotransformation was carried out at 37℃ in 8 L of reaction mixture containing 4% (W/V) D, L-p-HPH, 1% intact cell (wet weight) and 0.1 mol/L sodium phosphate (pH 9.0), with a molar yield of 96% within 5 hours. The pure product CpHPG was further chemically converted to D-p-HPG with an 89% yield. Thus, the overall conversion efficiency of D, L-p-HPH to D-p-HPG was 84%. The chemical purity and optical purity of D-p-HPG was 99% and 99.5%, respectively. The melting point (mp), optical rotation and infrared (IR) absorption spectrum of the product were identical with the control product of D-p-HPG.

Proteomic analyses is an effective way to identify protein constitutients in Lewy body-like inclusions (or aggresome) in vitro. Synthetic proteasome inhibitor (PSI, 10 µmol/L) exposure for 48 hours was used to induce the formation of cytoplasmic proteineous inclusions (termed as PSI-induced inclusions) in PC12 cells. The proteomic approaches of biochemical fractionation, two-dimensional electrophoresis (2-D) and identification via peptide mass fingerprints (PMF) were deployed and 20 protein components of LBs were identified, including 2 proteins involved in the production of synaptic neurotransmitter, 6 subunits of the 26S proteasome, 2 cytoskeleton proteins, 2 subunits of mitochondrial complexes, 1 anti-oxidant protein, and 7 chaperone or chaperone-like proteins. The results suggested that these LB protein components might had been recruited in PSI-induced inclusion formations in PC12 cells under the conditions of protease inhibition.

Changes of intracellular Ca 2+ ,pH value and mitochondria membrane potential(△Ψ m) in the apoptosis of MGC 803 cells induced by water soluble constituents of Glycyrrhiza uralensis Fisch(WSCG) were investigated.MGC 803 cells were incubated with 0.5,0.75,1.0 and 1.5 g/L WSCG for 1,4,7,11,15,19 and 23 hours respectively.The percentage of apoptosis and intracellular Ca 2+ content increased in a dose and time dependent manner,except that the intracellular Ca 2+ content of 1.5 g/L WSCG treated cells started to decrease after 11 hours.The cells were alkalized after various treatments,except that 1.5 g/L WSCG treated cells turned to acidify after 11 hours.It was found that the mitochondria △Ψ m of all treated cells decreased drastically at the first hour,then kept decreasing slowly until the 23rd hour.The results indicated that intracellular Ca 2+ ,pH and mitochondria △Ψ m all played pivotal roles in the apoptosis of MGC 803 cells induced by WSCG.

Nrf2(NF-E2-related factor 2)/ARE(antioxidant response element) is a novel defensive pathway involved in oxidative and chemical stress of cells. Underphysiological conditions, Nrf2 binds to kelch-like ECH-associated protein-1(Keap1) in the cytoplasm, remains inactivated and easy to be degradated. Under oxidative or chemical stress, Keap1 modification or Nrf2 phosphorylation result in activation of Nrf2 through its dissociation from Keap1.The activated Nrf2 translocates into the nucleus and interacts with ARE. The expression of cytoprotective target genes, including phase Ⅱdetoxifying enzymes, antioxidant proteins and the molecular proteasome/chaperones is then transactivated in response to the stress．Signal molecules, such as MAPK，PI3K/AKT，PKC, etc., extensively involved in the process of Nrf2 activation and translocation to the nucleus, however，which pathway to be activated and play a key role depends on the type of stimuli or cells and the way of stimulation applies．This article summarizes the latest research progresses regarding the molecular structures and the activation mechanisms of Nrf2，the signaling molecules and downstream genes of Nrf2/ARE pathway and their implications in tumor therapy，inflammation and aging．

Autophagy plays a critical role in replicative senescence cells. However, it remains poorly understood. Thereby we constructed the exogenous H2O2-nduced 2BS premature senescent cell model. And the β-Galactosidase (β-Gal) staining was conducted to confirm premature senescence. Morphological methods, specific markers for autophagy and mTOR signaling pathway were used to illuminate the change of autophagosomes. Monodansylcadaverine (MDC) staining and electron microscopy were firstly applied to detect the morphology of autophagosomes. GFP-LC3 was analyzed by fluorescence microscopy and immunoblotting was employedto examine the protein expression of LC3. Then, p70S6, the downstream effector of mTOR pathway, was examined by Western blotting. Consequently, we found that in the H2O2-induced premature senescent 2BS cell, autophagosomes increased in contrast to the young cells and have a protective function in premature senescence.

In human, over 70% of genome is continuously transcribed. In fact, only 1%～2% of the genome are coding genes, while 80%～90% non-coding regulatory elements are transcribed into long non-coding RNAs (LncRNA). Studies of LncRNAs have the potential to be more challenging than other RNAs. LncRNA tends to be poorly conserved interspecies and its expression often shows more cell type specific than protein coding genes. In addition, LncRNA also shows low expression levels. It is now well appreciated that LncRNA was involved in a wide variety of biological processes, such as epigenetics, cancer, and brain function. Current evidence suggests that LncRNA could regulate gene expression by diverse mechanisms (execute molecular functions as scaffold molecules, molecular guides, molecular decoys, and mediators on signaling pathway) and it may largely depend on the structure of LncRNA. Comprehensive discovery of the structure, function and mechanism of LncRNA not only provides a useful explanations of the physiological and pathological processes of the organism, but also gives us a new perspective to diagnosis, prevent and treatment some clinical diseases. Here we review the recent progress of the structure, function and underlying functional mechanisms of LncRNA.

Channelrhodopsin-2(ChR2), a directly light-activated cation-selective ion channel, is a seven-transmembrane (7-TM) helix protein isolated from Chlamydomonas reinhardtii, and contains the light-isomerizable chromophore all-trans-retinal. ChR2 opens rapidly after absorption of a photon to generate photocurrent. When it was expressed in excitable cells using a variety of transfection techniques, ChR2 makes depolarization of the excitable cells very straightforward after stimulated by blue light. It is useful for many bioengineering and neuroscience applications, such as control of neuronal firing for probing of neural circuits. Compared with the traditional electrophysiological technique and the pharmaceutical methods, ChR2 has the higher selectivity and specificity. The emerging field of controlling networks of genetically modified cells by light has been termed optogenetics. ChR2, a critical component of optogenetics, is a novel promising tool for neuroscience research. ChR2 has been applied to many aspects of nervous system research, such as visual neural circuit, tactual neural circuit, auditory neural circuit and olfactory neural circuit, as well as some nervous system disease research. The review highlights the recent advances of ChR2 in the study of nervous system, prospecting its future profile.

Gene silencing via microRNA(miRNA) expression and small interference RNA(siRNA) become important methods in for gene regulation. SiRNAbased RNA interference mediates the gene silencing by sequence complementary with target mRNA, whereas miRNA negatively regulates the gene expression at the posttranscriptional level through complementary sites at the 3′ untranslated regions of the target genes. The siRNA and miRNA regulation of gene expression has wide implications for the understanding of biology, as well as for disease etiology and treatment. Three sets of macromolecules, Dicers, Agos and 20 nt~25 nt RNA duplex, are introduced as the signature components of RNA silencing, two types of the classic silencing mechanisms of siRNA and miRNA are described. This review also compares the differences and discusses about the new arguments about the biological origins and silencing mechanisms of siRNA and miRNA.

Endoplasmic reticulum (ER) stress has been recognized as one of most important contributors to diabetes mellitus and neurodegenerative disorders. Homocysteine, oxidative stress and calcium metabolism disturbance were proved to induce endoplasmic reticulum stress cascade processes, including inhibition of protein synthesis initiation, expression of endoplasmic reticulum stress protein, and/or endoplasmic reticulum related apoptosis. Each of those dynamic processes can be biochemical discerned and dissected into unfolded protein response (UPR), integrated stress response (ISR), and/or endoplasmic reticulum associated death (ERAD) respectively. The cells will be eliminated by apoptosis via ERAD, once while their ER stress responses unfavorably go beyond UPR and ISR processes. A comprehensive understanding of the impact of the ER stress pathway within those suffering cells on their pathological process might be conducive to provide a new target for the prevention and treatment of those diseases.

DNA methylation plays a crucial role in various cellular functions such as differentiation, genomic stability, Xinactivation and imprinting. DNA methylation is mediated by DNA methyltransferases (DNMTs), including DNMT1, DNMT3a and DNMT3b. Aberrent DNA methylation has been observed in many types of cancer. Promoter hypermethylation causes silencing of key tumor supperssor genes, and consequently leads to cell proliferation, angiogenesis, invasion and metastasis. Genomewide hypomethylation contributes to genomic instability, which is an important feature of various malignancies. This review examined alteration in DNA methylation and the consequent effect on gene expression in cancer, and discussed the use of DNA methylation inhibitors in cancer therapy.

利用逆转录PCR和RACE技术获得建鲤2种Δ6脂肪酸去饱和酶（FADs6-a,FADs6-b）的全长cDNA序列. FADs6-a基因的cDNA总长为1 966 bp，开放阅读框为1 335 bp，编码444个氨基酸；FADs6-b基因的cDNA总长为1 931 bp，开放阅读框为1 335 bp，编码444个氨基酸.FADs6-a和FADs6-b基因都包括N端细胞色素b5结构域、3个富含组氨酸的结构域和2个推测的跨膜区，具有典型的Δ6脂肪酸去饱和酶结构特点.氨基酸同源性分析显示,建鲤FADs6-a和FADs6-b与斑马鱼的相似性较高，而与海水鱼类的相似性较低，与人类FADs6的相似性高于与FADs5的相似性.通过实时荧光定量PCR(RT-qPCR)检测该基因在建鲤幼鱼不同组织中的表达量, 发现2种Δ6 脂肪酸去饱和酶基因在建鲤肝脏的表达量最高，其次是肠、脑、肌肉、心和肾，而前肠高于后肠，FADs6-a的表达量高于FADs6-b.得到的结论是，建鲤具有2种类型的合成高度不饱和脂肪酸(HUFA)的关键酶—Δ6脂肪酸去饱和酶，在肝脏和肠道中的含量较多，且FADs6-a和FADs6-b在结构和组织的表达方面都存在差异，这为进一步研究建鲤HUFA的合成途径及调控机理奠定了基础.

 Carbohydrates have become a hotspot in the field of glycoscience due to their various bioactivities and important biological roles in the life activities. High sensitivity and high resolution micro-analytical techniques are required to elucidate the relationship between carbohydrate structure and their complicate biological functions. In recent years, several analytical technologies of labeling methods have been developed. The fluorescence labeling methods which improved the chromatographic determination and structural characterization have drawn wide attention. Both pre-column and post-column approaches have shown several advantages of high sensitivity, good resolution and versatility for labels. This paper compares the features and mechanisms of various fluorescent derivatization methods in applications for different carbohydrates of neutral, amino-, and acidic sugars, including sialic acid, uronic acid and sulfated sugars, as well as non-reducing sugars.

Recent studies showed that adipose triacylglyceride lipase (ATGL) functioned as an additional key enzyme for adipose lipid mobilization. ATGL specifically hydrolyzes the first ester bond of triglycerides and is a rate-limiting enzyme in triacylglycerol hydrolysis. The wide expression of ATGL suggested its important role in lipid metabolism in various tissue and cell types. The expression and activity of ATGL are regulated at both transcriptional and posttranslational levels. ATGL are also found to associate with obesity, diabetes, hepatic steatosis, and other related metabolic disorders. We focus on the discussion of structural characteristics, expression, biological functions, and regulatory mechanism of ATGL, and prospect on its potential application in the future.

Methylation at the 5 position of cytosine(C) in DNA is an important epigenetic modification involved in gene regulation, genomic imprinting, X-chromosome inactivation, suppression of repetitive elements, and carcinogenesis. The 5-methylcytosine (5mC) can be further converted to 5-hydroxymethylcytosine (5hmC) by TET (ten-eleven translocation) proteins family, which is essential for DNA demethylation. 5hmC is enriched at both the start sites of actively transcribed genes and extended promoter regions of Polycomb-repressed genes. TET proteins include three members TET1，TET2 and TET3, and belong to alpha-ketoglutaric acid (α-KG)and Fe²⁺-dependent dioxygenases in which catalysis is a oxidative process. Mouse Tet1 has a dual function in promoting transcription of pluripotency factors and participating in the repression of developmental regulators in embryonic stem cells. The disruptions of human TET proteins are associated with hematologic malignancies, such as frequently mutation of TET2 in myeloproliferative disorders/ neoplasms. These studies on TET proteins and 5hmC will provide new perspective for DNA methylation /demethylation and their biological function.

The endosomal sorting complex required for transport (ESCRT) system is essential for the degradation of ubiquitinated membrane proteins and comprises a major pathway for MVB (multi-vesicular body) formation. ESCRT is also involved in retrovirus budding, cell division, autophagy and fungi pH sensing. Five protein complexes were identified in the ESCR system: ESCRT-0, -Ⅰ, -Ⅱ,-Ⅲ and Vps4-Vta1. ESCRT-Ⅰand -Ⅱinduce the formation of the initial bud during the budding process. ESCRT-Ⅲ forms a lattice at the endosomal membrane and completes the scission at the bud neck, and Vps4Vta1 complex finally disassembles the ESCRT-Ⅲ lattice to recycle ESCRT machinery components. This review summarizes the structural features, budding mechanism and biological roles of the ESCRT system.

Reactive oxygen species (ROS) are highly reactive molecules produced in cell metabolism, thus make surrounding molecules vulnerable to oxidation. ROS are involved in various intracellular signal transduction pathways, and are believed to be important regulators of physiological processes. Autophagy is the degradation and recycling of components in eukaryotes through the lysosome system, and is a key player in cellular response to stress and the pathogenesis of related diseases. In this paper we reviewed the latest studies about ROS and autophagy-related regulation, focusing on autophagy modulation mediated by ROS as signal molecules, especially on signal transduction mechanisms involved in ROS-induced autophagic cell death, autophagic cell survival and mitophagy.

Cancer stem cells (CSCs), also known as tumor initiating cells, are a small subset of tumor cells with the biological characteristics that are similar to those of normal stem cell including self-renewal and differentiation. CSCs are important for cancer occurrence and development. The specific surface markers and abnormal active signal pathways is different from other cancer cells. Finding and identifying the specific surface markers and understanding the related regulatory mechanisms of CSCs behaviors is crucial to developing the new strategies for cancer targeted and personalized therapies in the future. To give a new insight aiming at antibody therapeutics that target CSCs,in this paper,we summarized the current known surface markers and signal pathways.

RNA silencing is a kind of cellular regulating system for mRNA cleavage or translation repression mediated by RNA induced silencing complex (RISC) under sequence-specific direction of small RNAs. As the core component of RISC, AGO(argonaute) protein consists of four domains: N terminus, PAZ, MID and PIWI. PAZ domain recognizes and binds 3-′ terminal 2nt overhang of small RNA duplex non-sequence-specifically. ‘Conserved pocket located at the interface between MID and PIWI recognizes and binds the first nucleotide of 5-′ terminus of small RNA. PIWI domain has a catalytic center for mRNA cleavage. According to the phylogenetic analysis, AGO family is divided into three groups: AGO-like, PIWI-like and GROUP3. Arabidopsis encodes ten AGO proteins (AtAGOs 1-10). Amongst, AtAGOs 1, 4 and 7 have been identified to possess cleavage acitivy, and to be involved in some small RNA pathways. In addition, parti ally functional redundancy exists between AtAGO1 and AtAGO10, AtAGO1 and AtAGO7, and AtAGO4 and AtAGO6.

Perilipin is one of the major members in PAT(perilipin, ADRP, TIP-47) proteinfamily, whose members not only have similar sequences but also have similar gene structure. It is a highly phosphorylated adipocyte protein which locates at the surface of the lipid droplet. It serves important functions in double-regulating triacylglycerol metabolism by blocking lipase approaching droplets to reduce rates of basal lipolysis and facilitate hormonally stimulated lipolysis, then regulates the storage and release of TG. Perilipin plays a important role in lipid metabolism. Its expression and regulation may be associated with obesity and related disorders such as diabetes, atheroma and insulin-resistance. This review mainly introduces the discovery, denomination, structual characteristic, function of perilipin, regulation of hormones and transcription factor on perilipin and its interaction with related lipase. Current researches focus on the interaction of perilipin and HSL, but no research on the interaction of perilipin and ATGL.

As a member of the Fox family, FoxO1 transcription factor is involved in the regulation of apoptosis, oxidative stress, DNA damage repair, cancer development, angiogenesis and glucose metabolism. The phosphatidylinositol 3kinase (PI3K) in the Akt pathway phosphorylates FoxO1 and mobilizes FoxO1 from the nuclei to the cytoplasm, leading to inactivation of FoxO1 and its impact on the downstream targets. Acetylation of FoxO1 attenuates its binding to the target DNA and enhances its phosphorylation, thereby decreases the transcription activities of FoxO1. This review focuses on the FoxO1 posttranslational modifications and their roles in the transcriptional regulation.

Reverse chromatin immunoprecipitation assay (reverse ChIP) is a new method to study DNA-protein interaction in vivo. It uses a specific nucleic acid probe to isolate genomic DNA with its associated proteins and identifies all the proteins bound to those loci using mass spectrometric analysis. This method allows the unbiased identification of targeted DNA locus-associated proteins, especially to find the regulatory proteins associated with the known DNA elements. Reverse ChIP will be of great value in the identification of targeted DNA locus-associated proteins and the study of DNA-protein interaction.

Recombinase polymerase amplification(RPA) is a kind of isothermal nucleic acid amplification technique rising in recent years. RPA is more rapid, convenient and efficient than the polymerase chain reaction (PCR) and other isothermal amplification techniques. In this paper, this new technique will be introduced in detail, with reviewing over the development of its relevant researches and applications until now on in medical diagnosis, agriculture, food safety, biological safety and so on. It is expected that this technique will attract more attention and be perfect in the near future making it play an important role in more fields, and what’s more, write a new start in nucleic acid detection.

Recently, long noncoding RNAs (lncRNAs), a significant new class of transcripts, have been aroused the concern in the scientific community. They were first revealed following the largescale sequencing of a fulllength cDNA library in mouse. LncRNAs, larger than 200 nucleotides, are lack of proteincoding function and have been found to be pervasively transcribed throughout eukaryotic genome. They account for the majority of the transcripts in mammalian genome. In comparison to small noncoding RNAs, the functions of lncRNAs are little understood. Fortunately, a growing number of studies have found that lncRNAs are involved in the multilevel regulation of gene expression. They play significant roles in the processes of embryonic development, the evolution of species, cell differentiation and diseases ranging from neurodegeneration to cancer. In this review, based on a brief introduction to lncRNAs, we have highlighted some recent progresses about their functions in the regulation of gene expression at transcriptional and posttranscriptional levels and epigenetics.

Most work of early proteomics was focused on the protein expression profile of cells on the different growth period or cells perturbed by mitogens or diseases. However, many vital processes are controlled not only by the relative abundance of proteins but also by reversible post translational modifications with specific spatial and temporal distributions. Consequently，it is important for comprehending complex biological functions of proteins to reveal the post- translational modification occurrence. Because of the low stoichiometry and broad dynamic range, research on the post-translationally modified proteins is still a big technical challenge. Nevertheless,development and integration of different methods, such as affinity-based enrichment methods,multi-dimensional separation technologies and mass spectrometry give the promise to probe the posttranslational proteomics in a large scale. Currently, there are several major kinds of large-scale post-translational modification study, of which phosphorylation and glycosylation are researched more frequently. This article reviews the strategies and techniques used in high-throughput investigation of post-translationally modified proteins in recent years.

The nuclear envelope is composed of two lipid bilayers, outer membrane and inner membrane. Nuclear pore complex (NPC) are formed at sites where the inner and outer membranes of the nuclear envelope are joined and it is a physical barrier of any passive and active transport between the cytoplasm and nucleus. Proteins functioned in the nucleus (used to be called as Cargoes) are synthesized in the cytoplasm and transported to the nucleus across nuclear pore complex in an energy-dependent way. This process is mainly regulated by nucleoporins in NPC, importins in cytoplasm and RanGTP. In addition, the structural character of cargo proteins plays essential roles in the transport to nucleus. This review mainly focuses on the molecular mechanism of protein transportion to the nucleus and recent progresses.

Metagenomics became a developing discipline for the study of microbial genes since 1990s. With metagenomic approaches, total DNA was directly extracted from the environment without the need of the microorganism cultures, therefore, to allow researchers to investigate at the scope of more than 99 % of the microbial genes in overall microorganisms. In general, metagenome technology can be applied in two aspects, to screen for functional genes and exploit proteins with new functions; or to find out the interactions among various microorganisms or between microorganisms and their surrounding environment. As the application of metagenomic technology expands, the researches on library construction, screening, sequencing and analysis advances to drive metagenomics towards even more practical values.

Signal transducers and activators of transcription 3 (STAT3), a transcription factor, plays key roles in regulating the signal transduction events mediated by cytokines and growth factors. In normal status, STAT3 was involved in many activities including survival, proliferation, differentiation and apoptosis. In addition, STAT3 was also be positively regulated by JAK/STAT, MAPK,mTOR pathway. Persistent activation of STAT3 has been demonstrated to be closely correlated to promotion of proliferation, anti-apoptosis, invasion, metastasis, angiogenesis and immune escaping behaviors in tumor cells. Therefore, strategies for inhibiting the roles of STAT3 signal pathway in tumor cells, including oligonucleotides, small molecular weight inhibitors and peptide aptamers, have been extensively studied. Current results showed that these strategies could inhibit tumor process and should be promising in antitumor therapy. In this study, functions of STAT3 and its potential applications in antitumor therapy would be reviewed.

Plasmid pKD46 can express three proteins: Gam, Bet and Exo. Gam inhibits the host RecBCD exonuclease V, so that Bet and Exo can gain access to DNA ends to promote recombination. BW25113 with pKD46 has the function of recombination when induced by L arabinose. PCR products were obtained by using primers with 50 bp extension which were homologous to clpP and by using template plasmid named pKD3 carrying chloramphenicol resistance gene flanked by FRT sites. The PCR products were introduced into BW25113 by electroporation. The strains expressing chloramphenicol resistance gene were selected by chloramphenicol agar. The chloramphenicol resistance gene was then eliminated by using a helper plasmid, pCP20, encoding the Flp recombinase.Using this system, ClpP gene in chromosome of Escherichia coli was deleted.