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Leptin is a peripheral satiety hormone that also plays important roles in energy homeostasis in vertebrates ranging from fish to mammals. In teleost fish, however, the regulatory mechanism for leptin gene expression still remains unclear. In this study, we found that glucagon, a key hormone in glucose homeostasis, was effective at elevating the leptin-AI and leptin-AII transcript levels in goldfish liver via both in vivo intraperitoneal injection and in vitro cells incubation approaches. The responses of leptin-AI and leptin-AII mRNA to glucagon treatment were highly comparable. In contrast, blockade of local glucagon action could reduce the basal and induced leptin-AI and leptin-AII mRNA expression. The stimulation of leptin levels by glucagon was caused by the activation of adenylate cyclase (AC)/cyclic-AMP (cAMP)/ protein kinase A (PKA), and probably cAMP response element-binding protein (CREB) cascades. Our study described the effect and signal transduction mechanism of glucagon on leptin gene expression in goldfish liver, and may also provide new insight into leptin as a mediator in the regulatory network of energy metabolism in the fish model.

In this paper ,we discus a wide class of loss functions for non-negative matrix factorization (NMF) and derive flexible and improved NMF algorithms based alpha-divergences for error measures, which generalize or combine several different criteria in order to extract intrusion signal.

Endophytic fungi are little known for their role in gibberellins (GAs) synthesis and abiotic stress resistance in crop plants. We isolated 10 endophytes from the roots of field-grown soybean and screened their culture filtrates (CF) on the GAs biosynthesis mutant rice line - Waito-C. CF bioassay showed that endophyte GMH-1B significantly promoted the growth of Waito-C compared with controls. GMH-1B was identified as Penicillium minioluteum LHL09 on the basis of ITS regions rDNA sequence homology and phylogenetic analyses. GC/MS-SIM analysis of CF of P. minioluteum revealed the presence of bioactive GA(4) and GA(7). In endophyte-soybean plant interaction, P. minioluteum association significantly promoted growth characteristics (shoot length, shoot fresh and dry biomasses, chlorophyll content, and leaf area) and nitrogen assimilation, with and without sodium chloride (NaCl)-induced salinity (70 and 140 mM) stress, as compared with control. Field-emission scanning electron microcopy showed active colonization of endophyte with host plants before and after stress treatments. In response to salinity stress, low endogenous abscisic acid and high salicylic acid accumulation in endophyte-associated plants elucidated the stress mitigation by P. minioluteum. The endophytic fungal symbiosis of P. minioluteum also increased the daidzein and genistein contents in the soybean as compared with control plants, under salt stress. Thus, P. minioluteum ameliorated the adverse effects of abiotic salinity stress and rescued soybean plant growth by influencing biosynthesis of the plant's hormones and flavonoids.

The beneficial effects of plant growth promoting fungi (PGPF) on plant growth and development are well documented. However, limited information is available on gibberellin (GA) production capacity of PGPF of endophytic origin. In current study, 11 fungal endophytes were isolated from cucumber roots and then screened on Waito-C rice, in order to identify plant growth promoting fungal strains. The fungal isolate GAH7 provided the maximum shoot length (11.3 cm) in comparison to control treatment (7.8 cm). In a separate experiment, bioassay of GAH7 significantly promoted growth attributes of cucumber. The GAH7 culture filtrate (CF) was found to contain physiologically active gibberellins in higher concentrations (GA1, 0.81 ng/ml; GA3, 4.34 ng/ml and GA4, 9.31 ng/ml) in conjunction with physiologically inactive GA9 (0.74 ng/ml), GA15 (0.97 ng/ml), GA19 (1.67 ng/ml) and GA20 (0.46 ng/ml). Isolate GAH7 produced higher amounts of GA3, GA4, GA9 and GA19 than wild type Fusarium fujikuroi, which was used as control for GA production. Gibberellins were analyzed through gas chromatograph/mass spectrometer (GC/MS) with selected ion monitoring (SIM). The fungal isolate GAH7 was later identified as a new strain of Phoma on the basis of sequence homology (99%) and phylogenetic analysis of 18S rDNA sequence.

Endophytic fungi are plant symbionts that produce a variety of beneficial metabolites for plant growth and protection against herbivory and pathogens. Fourteen fungal samples were isolated from the roots of soybean cultivar Daemangkong and screened on waito-c rice for their plant growth-promoting capacity. Twelve of the fungal isolates promoted plant growth, while two inhibited it. The fungal isolate DK-1-1 induced maximum plant growth in both waito-c rice and soybean. The plant growth promotion capacity of DK-1-1 was higher than the wild type Gibberella fujikuroi. Gibberellin (GA) analysis of culture filtrate of DK-1-1 showed the presence of higher amounts of bioactive GA3, GA4, and GA7 (6.62, 2.1 and 1.26 ng/mL, respectively) along with physiologically inactive GA5, GA15, GA19, and GA24. Phylogenetic analysis of 18S rDNA sequence identified the fungal isolate as a new strain of Cladosporium sphaerospermum. Gibberellin production and plant growth-promoting ability of genus Cladosporium are reported for the first time in the present study.

Based on the sequence of BAC (Bacterial Artificial Chromosome) along with the Cre/lox P system, the gene-targeting vectors to multiple loci of the repetitive internal transcribed spacers between rDNA genes in Leghorn chicken were constructed. The key material of multiple loci gene targeting in vivo would be obtained. First, the plasmid of pYLSV-TDN with TK, HRDS2, and Neo genes was constructed. The TK-HRDS2-Neo DNA fragment obtained from the plasmid of pYLSV-TDN was digested by Not I/HindIII and inserted into the upstream of the lox P site of BAC plasmid for obtaining the selective vector of BAC-TDN. The expression vector of pYLVS-GID with EGFP, hIFN genes, and HRDS1 was then obtained. The plasmid of BAC-TDN-VS-GID was obtained by cotransformation of the selective vector of BAC-TDN and the expression vector of pYLVS-GID to E. coli NS3529 through the action of Cre/lox P system. The gene-targeting vector of BAC-TDN-GID to multiple loci of the ITS region in Leghorn chicken was obtained by cleaving the sequence of pYLVS with the homing endonuclease of I -Sce I and ligating with the linker of LS. The insertion and the insert direction of DNA fragments were identified by restriction digestion or PCR and sequencing in each clone. The significance of the technique ofgene-targeting vector to multiple loci are shown as follows. First, the targeting loci were increased to 100 - 300. Second, the problems of unstable expression of inserted genes were partially solved. Third, the need for safety against toxicity integration was resolved. Fourth, the forbidden zone of gene integrating on the repetitive DNA sequences was broken through.

Seven sequence-specific ribozymes (M1GS RNAs) derived in vitro from the catalytic RNA subunit of Escherichia coli RNase P and targeting the mRNAs transcribed by the UL54 gene encoding the DNA polymerase of human cytomegalovirus were screened from 11 ribozymes that were designed based on four rules: (1) the NCCA-3' terminal must be unpaired with the substrate; (2) the guide sequence (GS) must be at least 12 nt in length; (3) the eighth nucleotide must be U, counting from the site -1; and (4) around the cleavage site, the sites -1/+1/+2 must be U/G/C or C/G/C. Further investigation of the factors affecting the cleavage effect and the optimal ratio for M1GS/substrate was carried out. It was determined that the optimal ratio for M1GS/substrate was 2:1 and too much M1GS led to substrate degrading. As indicated above, several M1GS that cleaved HCMV UL54 RNA segments in vitro were successfully designed and constructed. Our studies support the use of ribozyme M1GS as antisense molecules to silence HCMV mRNA in vitro, and using the selection procedure as a general approach for the engineering of RNase P ribozymes.

We have investigated the morphology and microstructure of carbon nanotubes and nanoparticles in cathode deposits prepared by self-sustained arc discharge. Scanning electron microscopy images indicate that there are two regions exhibiting different morphologies on the top surface of the cathode deposits. In the central region, there is a triangular pattern of spots with a diameter up to 100mum, which consists of carbon nanotubes and nanoparticles. In the fringe region, carbon nanotubes and nanoparticles are distributed randomly. In addition, carbon nanotubes in the central region have a larger inner diameter, compared with those in the fringe region. The outer diameter distribution of tubes in the central region is narrower than that of tubes in the fringe region, while the former has a smaller peak value than the latter. For the nanoparticles, they exhibit a different behaviour from the tubes existing in the same region. The difference between the microstructure of tubes or particles in the two regions is attributed to the different temperatures and temperature gradients during their formation.

Red light, temperature and gibberellins are well known for their capacity to induce higher germination in dormant seeds of several plant species. In the current study, we investigated the effect of various temperature (10, 13, 16, 19, 22 and 25°C) and gibberellic acid (GA 3 ) and GA 4+7 concentrations (0.1, 1, 10, and 100 uM) on germination of Poa annua, Alopecurus aequalis and Stellaria aquatica seeds incubated under continuous dark or treated with red light pulse of 80 mol m -2 s -1 red light (660 nm) for 10 min after 12 h of dark period. We found that incubation temperature and gibberellins (GAs) significantly enhanced seed germination in red light treated seeds, as maximum germination was observed for A. aequalis (67.3% at 13°C), P. annua (61.3% at 13°C), and S. aquatica (42% at 19°C) in such treatments. Exogenous GA 4+7 significantly promoted germination of P. annua and A. aequalis seeds, while GA 3 induced higher germination in S. aquatica seeds. It was observed that GAs were more effective in the presence of red light as least germination stimulation was recorded for GAs under dark condition and no germination was found in weed seeds at control and 0.1 mM GA 3 and GA 4+7 treatments. It can be concluded that lower temperature (13°C) and GA 4+7 in conjunctions with red light pulse, induced maximum germination in dormant weed seeds. Key words : Germination, temperature, nitrogen-containing compounds, red light, gibberellins.

Disseminated superficial actinic porokeratosis is an autosomal dominant cutaneous disorder characterized by many uniformly small, minimal, annular, anhidrotic, and keratotic lesions. The genetic basis for this disease is unknown. Using a genomewide search in a large Chinese family, we identified a locus at chromosome 12q23.2-24. 1 responsible for disseminated superficial actinic porokeratosis. The fine mapping study indicates that the disseminated superficial actinic porokeratosis gene is located within a 9.6 cM region between markers D12S1727 and D12S1605, with a maximum two-point LOD score of 20.53 (theta = 0.00) at D12S78. This is the first locus identified for a genetic disease where the major phenotype is porokeratosis. The study provides a map location for isolation of a gene causing disseminated superficial actinic porokeratosis.

In mammals, leptin is a peripheral satiety factor that inhibits feeding by regulating a variety of appetite-related hormones in the brain. However, most of the previous studies examining leptin in fish feeding were performed with mammalian leptins, which share very low sequence homologies with fish leptins. To elucidate the function and mechanism of endogenous fish leptins in feeding regulation, recombinant goldfish leptin-AI and leptin-AII were expressed in methylotrophic yeast and purified by immobilized metal ion affinity chromatography (IMAC). By intraperitoneal (IP) injection, both leptin-AI and leptin-AII were shown to inhibit the feeding behavior and to reduce the food consumption of goldfish in 2 h. In addition, co-treatment of leptin-AI or leptin-AII could block the feeding behavior and reduce the food consumption induced by neuropeptide Y (NPY) injection. High levels of leptin receptor (lepR) mRNA were detected in the hypothalamus, telencephalon, optic tectum and cerebellum of the goldfish brain. The appetite inhibitory effects of leptins were mediated by downregulating the mRNA levels of orexigenic NPY, agouti-related peptide (AgRP) and orexin and upregulating the mRNA levels of anorexigenic cocaine-amphetamine-regulated transcript (CART), cholecystokinin (CCK), melanin-concentrating hormone (MCH) and proopiomelanocortin (POMC) in different areas of the goldfish brain. Our study, as a whole, provides new insights into the functions and mechanisms of leptins in appetite control in a fish model.

As Chinese economy get fast developed, the residents' income improved greatly and the industry income gap varied constantly presenting a trend of continuous enlargement in the fluctuation.. This situation hinders the development of economy and the improvement of comprehensive strength. Thus to forecast the trend of continuous enlargement of the industry income gap has been extensively focused. With more complex objects and higher precision, there are some blind spots in the single model and it could not meet people's requirements. In order to have complementary advantages and get more accuracy in forecast, in this paper, we use the metabolism GM(1,1) Grey Forecasting Model to analyze the development trend and to forecast the income of the staffs among various industries (year 2011-2015) using the income data of the staffs among various industries of Chongqing City in recent years as Modeling Data, offering theoretical basis for making policy of shortening the income gap among various industries.

Different kinds of orbitozygomatic fractures lead to different levels of orbital structural destructions. Although the magnitude of the management of orbitozygomatic fractures varies considerably, an unsolved and important question remains regarding how to select the best surgical modality according to different kinds of fractures. Among 26 cases of unilateral noncomminuted orbitozygomatic fractures, a segmental osteotomy technique was used to repair the displaced orbitozygomatic complex. The fragment contributing to the orbital structure was mobilized and reduced, whereas the fragment without contribution to the orbital structure was not mobilized and left in situ. Next, the cuneate fragment was imbricated with the orbital osteotomy sites unilaterally or bilaterally. Miniplates and screws were used to span the osteotomy sites and provided fixation once the alignment of the orbit was achieved. The mean difference in volume between fractured orbits preoperatively and postoperatively was 2.47 mL. The mean difference in ocular globe projection was 2.76 mm. When postoperative results were compared with those of the preoperative site, statistically significant difference was noted. At follow-up 1 year postoperatively, the management of orbitozygomatic fractures using segmental osteotomy reduces complications and attains aesthetically satisfying results. Subjective assessment of the patients' globe position found that 88.5% of the patients were satisfied with the outcome, and 11.5% of the patients found it unacceptable. Compared with the traditional method, segmental osteotomy is a simple technique requiring less dissection and can reconstruct the orbital anatomic structure and restore globe position effectively.

To explore the toxicological mechanism of hydroquinone in human bronchial epithelial cells and to investigate whether DNA polymerase beta is involved in protecting cells from damage caused by hydroquinone.DNA polymerase beta knock-down cell line was established via RNA interference as an experimental group. Normal human bronchial epithelial cells and cells transfected with the empty vector of pEGFP-C1 were used as controls. Cells were treated with different concentrations of hydroquinone (ranged from 10 micromol/L to 120 micromol/L) for 4 hours. MTT assay and Comet assay [single-cell gel electrophoresis (SCGE)] were performed respectively to detect the toxicity of hydroquinone.MTT assay showed that DNA polymerase beta knock-down cells treated with different concentrations of hydroquinone had a lower absorbance value at 490 nm than the control cells in a dose-dependant manner. Comet assay revealed that different concentrations of hydroquinone caused more severe DNA damage in DNA polymerase beta knock-down cell line than in control cells and there was no significant difference in the two control groups.Hydroquinone has significant toxicity to human bronchial epithelial cells and causes DNA damage. DNA polymerase beta knock-down cell line appears more sensitive to hydroquinone than the control cells. The results suggest that DNA polymerase beta is involved in protecting cells from damage caused by hydroquinone.

Based on the sequence of BAC (Bacterial Artificial Chromosome) along with the Cre/lox P system, the gene-targeting vectors to multiple loci of the repetitive internal transcribed spacers between rDNA genes in Leghorn chicken were constructed. The key material of multiple loci gene targeting in vivo would be obtained. First, the plasmid of pYLSV-TDN with TK, HRDS2, and Neo genes was constructed. The TK-HRDS2-Neo DNA fragment obtained from the plasmid of pYLSV-TDN was digested by Not I/HindIII and inserted into the upstream of the lox P site of BAC plasmid for obtaining the selective vector of BAC-TDN. The expression vector of pYLVS-GID with EGFP, hIFN genes, and HRDS1 was then obtained. The plasmid of BAC-TDN-VS-GID was obtained by cotransformation of the selective vector of BAC-TDN and the expression vector of pYLVS-GID to E. coli NS3529 through the action of Cre/lox P system. The gene-targeting vector of BAC-TDN-GID to multiple loci of the ITS region in Leghorn chicken was obtained by cleaving the sequence of pYLVS with the homing endonuclease of I -Sce I and ligating with the linker of LS. The insertion and the insert direction of DNA fragments were identified by restriction digestion or PCR and sequencing in each clone. The significance of the technique ofgene-targeting vector to multiple loci are shown as follows. First, the targeting loci were increased to 100 - 300. Second, the problems of unstable expression of inserted genes were partially solved. Third, the need for safety against toxicity integration was resolved. Fourth, the forbidden zone of gene integrating on the repetitive DNA sequences was broken through.

A sequence-specific M1GS ribozyme (M1-T3) was constructed by covalently linking an oligonucleotide (guide sequence,GS) to the 3' terminus of M1 RNA ,the catalytic subunit of RNase P from Escherichia coli. The engineered ribozyme is targeted to the mRNA sequence encoding a protein kinase (UL97) of HCMV and could effectively cleave the mRNA segment in vitro. Further studies about the significance of some structural elements in the M1 GS (e.g. the 3' CCA tail sequence and a bridge sequence between the 3' terminus of M1 RNA and the 5' terminus of the GS) were carried out. The results showed that the bridge sequence of 88 nucleotides in a mutated M1 GS (i.e. M1-T3*) dramatically increased the cleavage activity to the substrate in vitro. Moreover, the 3'CCA tail sequence was confirmed to be a necessary element for the cleavage activity of M1 GS ribozyme. These data we got in the study will help in understanding the interaction between the M1 GS RNA and its substrate,and will markedly facilitate the research of a general gene targeting agent for anti-HCMV applications.

Mouse Pem, a homeobox gene, encodes a protein consisting of 210 amino acid residues. To study the function of mouse Pem protein, we used the yeast two-hybrid system to screen the library of 7-day mouse embryo with full-length mouse Pem cDNA. Fifty-two colonies were obtained after 1.57 x 10(8) colonies were screened by nutrition limitation and beta-galactosidase assay. Seven individual insert fragments were obtained from the library, and three of them were identified, one of which was confirmed to be the cell division cycle 37 (Cdc37) homolog gene by sequencing. The interaction between mouse Pem and Cdc37 homolog was then confirmed by glutathione S-transferase pull-down assay, and the possible interaction model was suggested.

To clone novel myelin protein related genes, two human ESTs, which shared significant similarity with the human myelin protein zero gene, were found by the comparison of homologue between the cDNA coding region sequences of MPZ gene and the EST database of NCBI. An 801 bp EST contig was assembled, which was 100% identical with a 128 kb genomic sequence, mapped to 1q24. A 435 bp open reading frame (ORF) within the 801 bp contig was shown by computer analysis. Two primers designed according to the sequence of the contig, were coupled with the primers(lambdagt10-5 and gt10-5) on the sequences flanking cloning site of the cDNA library vector to amplify the cDNA library sequences by nested PCR. New primers, designed based on novel cDNA sequences, were used for the PCR amplification with lambdagt10-5 and gt10-5 in the same way as above. Finally, the human myelin protein zero like gene isoform I and II (MPZL1a, MPZL1b GenBank AF095727, AF092424) were cloned. Comparison of gene and protein structures between MPZL1 and MPZ revealed that MPZL1 is the second member of MPZ family. Mutation analysis of MPZL1 gene was performed in 24 Charcot-Marie-Tooth disease (CMT) families and 26 nonsyndrome deafness families, but no mutation was found.