A large scale screen for genes （3rd chromosome） related to Wingless signaling pathway

A wing specific F 1 genetic screen was carried out using the powerful Drosophila genetic system, combined with yeast FRT/FLP and GAL4/UAS system. Form the wing phenotypes and germline clone embryonic cuticle phenotypes observed in these mutant alleles, a number of mutant alleles of known or unknown genes were isolated. Among them, fifteen mutant alleles related to Wingless signal transduction were further isolated; the arm of these mutations located were determined, and their location in the chromosome were roughly mapped.     

Enhancement of Au-Ag-Te contents in tellurium-bearing ore minerals via bioleaching

The purpose of this study was to enhance the content of valuable metals, such as Au, Ag, and Te, in tellurium-bearing minerals via bioleaching. The ore samples composed of invisible Au and Au paragenesis minerals (such as pyrite, chalcopyrite, sphalerite and galena) in combination with tellurium-bearing minerals (hessite, sylvanite and Tellurobismuthite) were studied. Indigenous microbes from mine drainage were isolated and identified as Acidithiobacillus ferrooxidans, which were used in bioleaching after adaption to copper. The effect of the microbial adaption on the bioleaching performance was then compared with the results produced by the non-adaptive process. The microbial adaption enhanced the Au-Ag-Te contents in biological leaching of tellurium-bearing ore minerals. This suggests that bioleaching with adapted microbes can be used both as a pretreatment and in the main recovery processes of valuable metals.     

Microbial community structure in major habitats above 6000 m on Mount Everest

Bacterial abundance in surface snow between 6600 and 8000 m a.s.l. on the northern slope of Mt. Ev- erest was investigated by flow cytometry. Bacterial diversity in serac ice at 6000 m a.s.l., glacier melt- water at 6350 m, and surface snow at 6600 m a.s.l. was examined by constructing a 16S rRNA gene clone library. Bacterial abundance in snow was higher than that in the Antarctic but similar to other mountain regions in the world. Bacterial abundance in surface snow increased with altitude but showed no correlation with chemical parameters. Bacteria in the cryosphere on Mt. Everest were closely related to those isolated from soil, aquatic environments, plants, animals, humans and other frozen environ- ments. Bacterial community structures in major habitats above 6000 m were variable. The Cyto- phaga-Flavobacterium-Bacteroides (CFB) group absolutely dominated in glacial meltwater, while β-Proteobacteria and the CFB group dominated in serac ice, and β-Proteobacteria and Actinobacteria dominated in surface snow. The remarkable differences among the habitats were most likely due to the bacterial post-deposition changes during acclimation processes.     

Sponge fossil assemblage from the Early Cambrian Hetang Formation in southern Anhui

Abundant well-preserved large articulated sponge fossils and isolated spicules have been reported from the Early Cambrian Hetang Formation, southern Anhui Province. This unique epifaunal fossil assemblage dominated by articulated sponge fossils is called the Xidi Sponge Fauna. The sponge fauna lived in a quiet oxygenic environment below the storm wave base. Bloom of phytoplankton and rapid sedimentation rate resulted in the deposition of the black shales. Sufficient food supply, lack of other competitors, abundant ecological niches, and demand for oxygen during early Cambrian were in favor of the diversification and evolution of large sponges in the Early Cambrian.     

Self-assembly of insulated molecular wires of a watersoluble cationic PPV and anionic dendrons

Insulated molecular wires of poly（phenylenevinylene） （PPV） were prepared by wrapping the conjugated backbones with dendrons through a noncovalent approach. It was found that electrostatic interaction between the quaternary ammonium groups of PPV-1 and the carboxylate moieties in dendrons induced the packing of dendrons along PPV-1 conjugated backbones. Absorption and emission spectroscopic examinations in solution and solid film indicated that the PPV-1 backbones adopted a more planar and isolated conformation in the complexes. Furthermore, interchain interactions in the complexes could be greatly reduced, improving the quantum yield of PPV-1.     

Several oxygenated monoterpenes from petals of Rosa damascena

Aim To discover the oxygenated monoterpenes that play a critical role in the biosynthetic pathway of monoterpene in Rosa damascene.Method Several kinds of isolation techniques such as liquid-liquid partition,silica column chromatography,thin layer chromatography and high performance liquid chromatography were used to separate the compounds and several identification techniques such as GC-MS and 1H-NMR were used to determine the structure of the isolated compounds.Results Three oxidized acyclic monoterpenes were obtained.They are 8-hydroxygeraniol,8-hydroxynerol and 8-hydroxylinalool.Conclusion The compounds are the important materials of pharmaceutical,cosmetic and food industries.The work has not only provided a new method for the determination of the main aromatic components of Rosa damascena,but also built up an important foundation for the study of biosynthesis of monoterpenoids in the plant.     

Isolation of a pollen specific promoter in tritordeum

The promoter is a cis-acting element in regulating gene expression. A promoterless plasmid containing UidA gene was transformed into tritordeum by barmbadment. Histochemical analysis of various tissues in transgenic tritordeum was carried to examine tissue-specific expression of GUS(beta-glucuronidase) activity. The pollen-specific promoter was trapped and identified successfully in a transformant line. PCR(polymerase chain reaction) method was used to isolate this pollen-specific promoter. By sequencing and analyzing the amplified fragment from PCR, a part of UidA gene and a flanking sequence were obtained. Some essential elements of plant promoters were found in the sequence. To determine the function of it, the cloned fragment was fused with UidA gene, then cloned and transformed into Triticum durum. The transgenic plant transformed by this vector showed GUS expression only in pollen. Therefore a pollen-specific promoter was isolated successfully.     

A novel peptide,selected from phage display library of random peptides,can efficiently target into human breast cancer cell

To develop a targeting vector for breast cancer biotherapy, MDA-MB-231 cell, a human breast cancer cell line, was co-cultured with pC89 （9 aa） phage display library of random peptides. In multiple inde-pendent peptide-presenting phage screening trials, subtilisin was used as a protease to inactivate extra-cellular phages. The internalized phages were collected by cell lysising and amplified in E. coli XLI-Blue. Through five rounds of selection, the pepUde-presenting phages which could be internalized in MDA-MB-231 cells were isolated. A comparison was made between internalization capacities of peptide-presenting phages isolated from MDA-MB-231 cells and RGD-integrin binding phage by coculturing them with other human tumor cell lines and normal cells. The nucleoUde sequences of isolated peptide-presenting phages were then determined by DNA sequencing. To uncover whether phage coat protein or amino acid order was required for the character of the pepUde to MDA-MB-231 cells, three peptides were synthesized. They are CASPSGALRSC, ASPSGALRS and CGVIFDHSVPC （the shifted sequence of CASPSGALRSC）, and after coculturing them with different cell lines, their targeting capacities to MDA-MB-231 cells were detected. These data suggested that the internalization process was highly selective, and capable of capturing a specific peptide from parent peptide variants. Moreover, the targeting internalization event of pepUdes was an amino acid sequence dependent manner. The results demonstrated the feasibility of using phage display library of random peptides to develop new targeting system for intracellular delivery of macromolecules, and the peptide we obtained might be modified as a targeting vector for breast cancer gene therapy.     

Sequence analysis of the Microcystis aeruginosa FACHB-912 phytochrome gene supports positive selection in cyanobacteria

The phytochrome gene from Microcystis aeruginosa FACHB-912,isolated from algal blooms in Taihu Lake in China,was sequenced and found to code for a wavelength transition light receptor protein.The gene was composed of five distinct domains:a PAS fold domain,a GAF domain,a phytochrome domain,a His Kinase A(phosphoacceptor) domain,and a histidine kinase domain.Red-absorbing phytochrome,far-red-absorbing phytochrome,and photoconversion kinetics were monitored via the spectral characteristics of the protein.To detect positive selection,homologous sequences from cyanobacterium phytochrome genes were obtained from GenBank.Potential selective pressure was identified by phylogenetic analysis with maximum likelihood analyses.The frequency of the phytochrome gene being subjected to selective constraints suggests that positive selection is a potentially important mechanism that promotes the evolution of the cyanobacteria phytochrome gene.The sites identified in this study provide targets for further research on the structural-functional role of these residues,and on the correlation with the mechanism of algal blooms.     

Pancreastatin, a novel pancreatic peptide that inhibits insulin secretion

In mammalian tissues the C-terminal amide structure has been found to occur only in neuroactive or hormonally-active peptides. About half known neuropeptide and peptide hormones have this unique chemical feature. Using a chemical detection method, a search for previously unknown peptides that possess the C-terminal amide structure in extracts of brain and intestine was carried out and a number of novel neuropeptides and hormonal peptides, designated neuropeptide Y, PHI, peptide YY, galanin and neuropeptide K were isolated. We recently performed a similar search in porcine pancreas and found a high concentration of a peptide having a glycine amide at its C-terminus. Here we report the isolation, primary structure and biological activity of this novel peptide. The 49-residue peptide strongly inhibits glucose-induced insulin release from the isolated perfused pancreas and was therefore named pancreastatin. It may be important in the regulation of insulin secretion and in the pathogenesis and treatment of diabetes mellitus.     

cDNA cloning of bovine substance-K receptor through oocyte expression system

The neuropeptide receptors which are present in very small quantities in the cell and are embedded tightly in the plasma membrane have not been well characterized. Mammals contain three distinct tachykinin neuropeptides, substance P, substance K and neuromedin K, and it has been suggested that there are multiple tachykinin receptors. By electrophysiological measurement, we have previously shown that Xenopus oocytes injected with brain and stomach mRNAs faithfully express mammalian substance-P and substance-K receptors, respectively. Here we report the isolation of the cDNA clone for bovine substance-K receptor (SKR) by extending this method to develop a new cloning strategy. We constructed a stomach cDNA library with a cloning vector that allowed in vitro synthesis of mRNAs and then identified a particular cDNA clone by testing for receptor expression following injection of the mRNAs synthesized in vitro into the oocyte system. Because oocytes injected with exogenous mRNAs can express numerous receptors and channels, our new strategy will be applicable in the general molecular cloning of these proteins. The result provides the first indication that the neuropeptide receptor has sequence similarity with rhodopsin-type receptors (the G-protein-coupled receptor family) and thus possesses multiple membrane-spanning domains.     

Leptin-regulated endocannabinoids are involved in maintaining food intake

Leptin is the primary signal through which the hypothalamus senses nutritional state and modulates food intake and energy balance. Leptin reduces food intake by upregulating anorexigenic (appetite-reducing) neuropeptides, such as alpha-melanocyte-stimulating hormone, and downregulating orexigenic (appetite-stimulating) factors, primarily neuropeptide Y. Genetic defects in anorexigenic signalling, such as mutations in the melanocortin-4 (ref. 5) or leptin receptors, cause obesity. However, alternative orexigenic pathways maintain food intake in mice deficient in neuropeptide Y. CB1 cannabinoid receptors and the endocannabinoids anandamide and 2-arachidonoyl glycerol are present in the hypothalamus, and marijuana and anandamide stimulate food intake. Here we show that following temporary food restriction, CB1 receptor knockout mice eat less than their wild-type littermates, and the CB1 antagonist SR141716A reduces food intake in wild-type but not knockout mice. Furthermore, defective leptin signalling is associated with elevated hypothalamic, but not cerebellar, levels of endocannabinoids in obese db/db and ob/ob mice and Zucker rats. Acute leptin treatment of normal rats and ob/ob mice reduces anandamide and 2-arachidonoyl glycerol in the hypothalamus. These findings indicate that endocannabinoids in the hypothalamus may tonically activate CB1 receptors to maintain food intake and form part of the neural circuitry regulated by leptin.     

Neuropeptides modulate the beta-adrenergic response of purified astrocytes in vitro

Neuropeptides may have functions in the central nervous system (CNS) other than altering neuronal excitability. For example, they may act as regulators of brain metabolism by affecting glycogenolysis. Since it has been suggested that glial cells might provide metabolic support for neuronal activity, they may well be one of the targets for neuropeptide regulation of metabolism. Consistent with this view are reports that peptide-containing nerve terminals have been seen apposed to astrocytes, but it is also quite possible that peptides could act at sites lacking morphological specialization. Primary cultures containing CNS glial cells have been shown to respond to beta-adrenergic agonists with an increase in cyclic AMP and, as a result, with an increase in glycogenolysis and have also been shown to respond to a variety of peptides with changes in cyclic AMP. In the study reported here, we have examined the effects of several peptides on relatively pure cultures of rat astrocytes. We demonstrate that the increase in intracellular cyclic AMP induced by noradrenaline is markedly enhanced by somatostatin and substance P and is inhibited by enkephalin, even though these peptides on their own have little or no effect on the basal levels of cyclic AMP. Vasoactive intestinal peptide (VIP) on the other hand increases cyclic AMP in the absence of noradrenaline. These results suggest that neuropeptides influence glial cells as well as neurones in the CNS and, in the case of somatostatin and substance P, provide further examples of neuropeptides modulating the response to another chemical signal without having a detectable action on their own.     

几种神经肽对生殖的影响和作用

关于神经内分泌对生殖调控的研究已越来越多地集中在神经肽的作用上.速激肽即是神经 肽中较大的一族.对从无脊椎动物及脊椎动物中发现的几种速激肽和一些重要的神经肽在生殖中 的影响作出简要综述.     

A novel active pentapeptide from chicken brain identified by antibodies to FMRFamide

The tetrapeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) and peptides structurally related to it, have been isolated from molluscan ganglia. They have widespread actions on both invertebrate and vertebrate tissues and there is increasing evidence that they are an important group of invertebrate peptide neurotransmitters. It is of interest that the primary amino acid sequence of FMRFamide forms the C-terminal tetrapeptide of an enkephalin-like heptapeptide (Met-enkephalin-ArgPhe) isolated from bovine adrenal medulla and striatum. Antisera to FMRFamide have been shown to react in radioimmunoassay and immunohistochemistry with material in the central nervous system of various vertebrate species, but the identity of this material, and in particular its relationship to the opioid heptapeptide, remains uncertain. We have used antibodies specific for the C-terminus of FMRFamide in radioimmunoassays to monitor purification of the material in chicken brain. We describe here the sequence of one of the peptides obtained. It is a biologically active peptide which does not seem to be related to other known vertebrate neuropeptides.     

Enkephalin-, VIP- and substance P-like immunoreactivity in the carotid body

The carotid body type I cell contains amines and has features, both morphological and cytochemical, which indicate that it may also produce a peptide. Many regulatory peptides are now known to be present in both central and peripheral tissues. In the periphery these neuropeptides occur in both classical endocrine (APUD) cells and the neurones of the autonomic nervous system. We have now investigated the possible presence of neuropeptides in the cat carotid body using both immunocytochemistry and radioimmunoassay. Met- and Leu-enkephalin-like material occurred in considerable quantities in carotid body extracts and enkephalin-like immunoreactivity was localised in type I cells. Both vasoactive intestinal polypeptide (VIP)- and substance P-like immunoreactivity was also present but was localised in nerve fibres distributed throughout the organ. These active neuropeptides are widely distributed in mammalian tissues, forming a diffuse regulatory system which now seems to include the carotid body.     

P物质对大鼠星状神经节细胞的除极化

应用细胞内生物电记录技术,观察神经肽P物质(SP)对大鼠星状神经节细胞的影响.SP在1 μmol～10 μmol或更高的浓度范围内,供试35个细胞, 有28个细胞发生膜除极反应.用低钙(0.25 mm)或用含河豚毒素(TTX,1 μmol)克氏液灌流神经节,不影响SP引起的除极反应的幅度和时程.SP引起除极反应的同时常伴有膜电阻增大.当膜电位增大时,除极化反应幅度变小,反转电位为-80 mV至-100 mV.研究表明,SP对部分星状神经节细胞具有兴奋作用,使通过这些细胞的信息传递增强;SP对细胞膜的除极作用是由于其引起细胞膜钾导降低所致.     

Nerve growth factor regulates expression of neuropeptide genes in adult sensory neurons

Nerve growth factor (NGF) is a trophic molecule essential for the survival of sympathetic and sensory neurons during ontogeny. The extent to which NGF is involved in the maintenance or regulation of the differentiated phenotypes of mature peripheral neurons is much less clear, however. Biochemical analysis of the actions of NGF upon peripheral neurons has been hampered by the lack of a preparation of neuronal cells that are responsive to NGF but do not require it for survival. We report here that in adult dorsal root ganglion neurons, which can be isolated, enriched and maintained in culture in the absence of neuronal growth factors, the expression of mRNAs encoding the precursors of two neuropeptides, substance P and calcitonin gene-related peptide is regulated by NGF. Our results provide the first direct evidence of a continuous dynamic role for NGF in regulation of peptide neurotransmitter/neuromodulator levels in mature sensory neurons.     

神经肽、神经递质与肿瘤的关系

对神经肽及神经递质对肿瘤增殖和分化的调控作用、调控机理及肿瘤细胞表面的神经肽受体的研究进展进行了综述。