Abstract:Essential tremor is one of the most common neurological disorders which the causes remain unknown. The clinical feature is heterogeneous and many ET patients have positive family history. Thus far, three gene loci have been identified and two susceptibility genes including DRD3 (the dopamine receptor D3 gene) and LINGO1 (the Leucine-rich repeat-and lg domain-containing NOGO receptor-interacting protein 1 gene) have been reported recently. The genetics of essential tremor will be summarized.

Abstract:Typically a drug target is a key molecule involved in a particular metabolic or signaling pathway, that is specific to a disease condition or pathology. Drugs may be designed that bind to the active region and inhibit this key molecule. Determining specific disease-related target molecules is the basis of modern drug development. In the process of drug target discovery, bioinformatics methods play irreplaceable roles, especially suited for the analyses of large-scale and multi-omics data. On current, many disease-related database resources have emerged. Various bioinformatics methods have been established based on biological network characteristics, multiple gene chips, proteomics and metabolomics data to discover potential drug targets, and predict the target druggability and side effects of drugs.

Abstract:Inflammation is the normal host tissue response to infection and injury. However uncontrolled and unresolved inflammation contributes to a range of acute and chronic human disease. Recent results have demonstrated several new families of lipid mediators, derived from eicosapentaenoic acid(EPA) and docosahexaenoic acid(DHA) of n-3 serial fatty acid, that including resolvins and protectins, to be both anti-inflammation and proresolving. Biosynthesis, anti-inflammatory action and mechanism of these new proresolving lipid mediators were described in order to complete the anti-inflammatory mechanism of n-3 polyunsaturated fatty acid and offer exciting new potential for therapeutic control.

Abstract:Cellulosomes produced by certain anaerobic microbes are commonly assembled from multiple subunits to macromolecular machines, and are extracellular protein complexes, which could organize and coordinate a variety of enzymic components to synergistically and effectively degrade lignocelluloses. Cellulosomes are the main forces of anaerobic hydrolytic celluloses, and play a vital role in breaking and using crystalline celluloses. The highly-efficient degradation on lignocelluloses of cellulosomes come from their self-assembled complex high-level structures, and the complex structures of the different anaerobic microorganisms have the astonishing diversity. The research advances on the diversity of assembly mode, structure, and the artificial design of cellulosomes were introduced.

Abstract:Calmodulin (CaM) is an important signal protein in eukaryotes and play multiple regulatory roles in eukaryotes' development in normal and stress conditions. Previous reports showed that calmodulin could accelerate animals and plants' cell proliferation in vitro. However, there is nearly no reports on cell proliferation in vivo in plants under normal and stress conditions. Here, calmodulin antagonist W7 inhibited the root elongation of Arabidopsis seedlings, and the area and cell number of root apical meristem were reduced, the expression of CYCB1;1 gene which plays a key role in cell proliferation was seriously restrained. This data indicates that activated calmodulin is required for cell proliferation. Abscisic acid (ABA), as a key hormone mediating plant adaptation to various environmental challenges, regulates many processes of plant growth and development, such as seed germination and seedling growth. In the present of W7, wild type arabidopsis seedlings are insensitive to ABA, and cam2cam3cam4 triple mutant had decreased sensitvity to ABA. The results implied that CaM might play important role in cell proliferation of root apical meristem and involved in ABA response.

Abstract:In order to explore the effect of exposure to subtoxic dose of organophosphorus insecticide chlorpyrifos on the formation of atherosclerosis induced by the high fat diet in New Zealand rabbits and analyze the possible mechanisms, thirty two healthy male New Zealand rabbits were divided randomly into four groups: control, high-fat diet, chlorpyrifos and high-fat diet + chlorpyrifos group. Subtoxic dose of chlorpyrifos (20 mg/kg·d) was administered by lavage every day for six months. The levels of serum fat, activities of cholinesterase and alanine aminotransferase, serum creatinine and blood urea nitrogen were measured respectively. The peritoneal macrophages were assembled and cellular cholesterol efflux was analyzed. Area of atherosclerosis plaque of thoracic aorta was measured by Sudan Ⅳ. Common carotid artery was fixed in formalin, sliced and HE dyed and pathology analysis system was used. The expression of ABCA1 was detected by Real-time PCR and Western blot. Compared with control group, serum TC, LDL and TG were singificantly increased and expressions of ABCA1 in liver, aorta and peritoneal macrophages were markedly increased and cholesterol efflux of peritoneal macrophages was significantly increased in high-fat diet group. There was obvious atherosclerosis lesion in thoracic aorta and common carotid artery in high-fat diet group. Compared with control group, activity of cholinesterase was singificantly decreased, but there were no symptom of intoxation and injury of function of liver and kidney, and level of HDL, expression of ABCA1 and cholesterol efflux were markedly decreased in chlorpyrifos group. Compared with high-fat diet group, activity of cholinesterase was decreased, but there were no symptom of intoxation and injury of function of liver and kidney, and expression of ABCA1 and cholesterol efflux were singificantly decreased, the atherosclerosis lesion areas in thoracic aorta and common carotid artery were increased in high-fat diet + chlorpyrifos group. These results suggest that a long time exposure to subtoxic dose of chlorpyrifos may accelerate formation of atherosclerosis induced by high fat diet in New Zealand rabbits, which the mechanism may be related to the decrease of ABCA1 expression and cholesterol efflux induced by chlorpyrifos.

Abstract:RING1 can bind to DNA and inhibit some gene transcription. RING1 was identified to interact with lamin A by screening a human skeletal muscle library in a yeast two-hybrid interacting screen. The results of one to one reverse hybridization indicated that AH109 transfected with RING1 and lamin A constructs could grow on SD/-Leu/-Trp/-Ade/-His medium. RING1 was inserted into pEGFP-N1 vector and the constructs including pDsRed-LA and pEGFP-RING1 was used to transfect HEK293 cells. Visualized by laser confocal microscopy, RING1 and Lamin A were co-located around nucleus. Interatcion of RING1 and lamin A was comfirmed by immunoprecipitations. These results showed a new lamin A binding protein that can present some evidence for lamin A acting on gene expression and cell senescence.

Abstract:The full length cDNAs of rotavirus structural protein genes, vp4, vp6 and vp7 were cloned from the rotavirus infected child stool specimen in Beijing through RT-PCR．The protein sequences and their antigenic determinants were predicted．According to the epitope peptide sequences, 4 epitopes from these structural proteins were chosen, a DNA fragment encoding all these epitopes was synthesized and cloned into the prokaryotic expression vector. Multiple epitope protein (rotavirus multiple epitopes, RME) expressed in E. coli can be recognized by the polyclonal antibody of rotavirus, and induce immune response in mice．The specific antibody IgG induced by RME can recognize human rotavirus (Wa strain), RME itself as well as individual epitope peptides. The antibody titer of IgG to RME is high (1∶40 000), while the titers to EV4, EV6 or EV7 are in a range of 1∶10 000 to 1∶20 000. However the IgG titer to the Wa strain is lower, i.e., 1∶2 500．Intriguingly, the RME-induced IgG can neutralize the Wa strain rotavirus challenge in the MAC145 cell line. This research has laid the foundations of producing effective bioengineering vaccines to rotavirus.

Abstract:von Willebrand factor (vWF) is a huge multimeric plasma glycoprotein with important functions involved in thrombosis and hemostasis. Its qualitative and/or quantitative abnormalities result in a bleeding disorder termed von Willebrand disease (VWD). Gene therapy is favorable for treatment of VWD because this disease is monogenic and the vWF is a secretory protein making non-specific targeting organ required for gene delivery. But the vWF gene is hardly packaged in most existing viral vectors especially the desired adeno- associated virus (AAV) vectors for its oversized cDNA in size (8.4 kb). The intein-mediated protein trans-splicing was explored to co-transfer split three fragmented vWF gene into eukaryotic cells by a ternary-vector system and the functional vWF protein was expected to be formed posttranslationally by protein trans-splicing. The vWF cDNA was broken into three fragments before codons of Cys¹⁰⁹⁹ and Ser²⁰⁰⁴ which required for protein splicing and then fused with Ssp DnaE and Ssp DnaB inteins respectively. A group of three eukaryotic expression vectors were produced by inserting these three fusion genes into pcDNA3.1(+) respectively. By transient co-transfection of 293 cells with these three vectors the conditioned culture supernatant was observed for vWF multimer pattern by electrophoresis, analyzed for vWF antigen and binding capacity of coagulation factor Ⅷ (FⅧ) quantitatively. With FⅧ gene co-transfection, the antigen and activity of FⅧ in the culture supernatant were measured. The data showed that with intein-mediated protein trans-splicing after translation the culture supernatant from cells co-transfected with intein-fused three fragmented vWF genes displayed a vWF multimer pattern and FⅧ binding capacity similar to normal human plasma and cells of vWF gene transfected positive control, and the FⅧ secretion and activity were increased dramatically in FⅧ gene co-transfected cells indicating the functional recovery of spliced vWF as a FⅧ carrier. It suggests that inteins could be used as a powerful means for tri-fragmental co-delivery of the vWF gene and may be valuable for application of intein-based ternary AAV vector in split vWF gene delivery in gene therapy for VWD to overcome the packaging limitation of AAV vectors.

Abstract:Chalcone synthase (CHS), coded by the chs gene super-family, is a key enzyme in flavonoid biosynthesis. Two independent promoters was isolated for chsA, named PchsA-L (550 bp) and PchsA-S (354 bp) (GenBank accession number EF199747 and EF199748 respectively), from the genomic DNA of Petunia hybrida. PchsA-L differs with PchsA-S mainly in that PchsA-L has a 182 bp fragment from 88～269 bp, and the sequence 103～201 bp has the characteristics of a typical intron. Both promoter sequences contain conserved sequences of TATA box, CCAAT box, cap site (CCATAA), and the flower-specific promoter sequences TACPyAT box, anther box (TAGAAGTGACAGAAAT), G-box (CACGTG), box1 element (ATGTCACGTGCCATC) and box2 element (TGTGTTGAAGGTTTGCTA). The petunia plant used for promoter cloning was a diploid with 14 chromosomes. Southern blotting showed that both promoters had multiple copies in the genome. The two promoters segregated in the offspring but the segregation did not meet the ratio of 1∶2∶1. qRT-PCR analysis showed no significant difference in chsA gene expression in non-UV-treated and UV-treated floral organ of plants with one or both promoters. The expression of chsA in the UV-treated seedling leaves was increased compared with UV-treated floral organ, and PchsA-L-driven chsA expression in the UV-treated seedling leaves was very significantly increased than that driven by PchsA-S while there was no chsA gene expression in non-UV-treated seedling leaves. The results show the presence of two independent promoters PchsA-L and PchsA-S for chsA in the petunia genome; the 182 bp intron-like sequence in PchsA-L promoter could significantly increased chsA gene expression in UV-treated seedling leaves.

Abstract:Thyroid hormone responsive spot 14 (THRSP) is a nucleoprotein induced by thyroid hormone, which makes an essential effect on the regulation of animal lipogenesis. In order to clarify the genetic mechanisms of the differences between Chinese native pig breeds (fat-type group, FTG) and introduced pig species (lean-type group, LTG) in carcass fat deposition, DNA samples of ear tissues were extracted from FTG(n = 228) composed of Wannan Spotted, Jixi Black and Dingyuan and LTG (n = 92) from Landrace, Large White, Duroc and their hybrid pigs and then detected the SNPs in the coding region of THRSP gene. The effects of SNPs on mRNA folding and protein secondary structure in FTPG and LTPG were predicted using Vienna RNA folding software and YASPIN web services, and their distribution and association with lipogenesis capability in FTG and LTG were analyzed. The results showed that the homologies of nucleotide sequence in coding region of pig THRSP gene with human and cattle were 86% and 88%, respectively. Two SNP sites, G123A and A308G, were located at 123 bp and 308 bp away from the start of the CDS, respectively. The mutation at G123A site was a synonymous mutation, but the mutation at A308G site caused the change of amino acids from lysine to arginine at 103rd site of THRSP protein which resulted in the changes of casein kinase Ⅱ phosphorylation site from TKEE to TREE, mRNA folding and protein secondary structure. The frequencies of alleles 123G and 308A were 0.975 9 and 0.589 9 in FTG, repectively, while that of alleles 123G and 308A were 0.657 7 and 0.815 2 in LTG. The total frequency of gamete 123G308A and 123G308G in FTG achieved 0.975 9, while that of gamete 123A308A and 123G308A in LTG was 0.815 3. Data from the present study imply the association of polymorphism at G123A and A308G sites with lipogenesis capability of pigs and their important regulating role in the expression of lipogenesis genes in pigs.