Molecular characterization and functional expression of opioid receptor-libe_1 receptor

The opioid receptor-libel receptor (ORL), an orphan receptor whose human and murine complementary DNAs,has been characterized recently. ORL transcripts are particularly abundant in the central nervous system. We demonstrated that ORL expressed in human neuroblastoma SK-N-SH and SH-SY5Y cell lines by radioligand binding assay, reverse transcription polymerase chain reaction (RT-PCR) and Northern analysis in the present study. Stimulation with ORL1 specific agonist, nociceptin/orphanin Fo, increased [34S]GTPrγS binding to SK-N-SH cell membranes (EC50 = 14 ±0.45 nM), and attenuated forskolin-stimulated accumulation of cellular cAMP (EC50= 0.80 ±0.45 nM, indicative that activation of ORL1 activates G proteins and inhibits adenylyl cyclase. Activation of ORL1 receptor was also accessed using CHO:hORL1 cell line by microphysiometer. Treatment of nociceptin/orphanin FQ increased extracellular acidification rate significantly.

Expression pattern and function analyses of the MADS thranscription factor genes in wheat(Triticum aestivum L.) under phosphorusstarvation condition

supported by the National Natural Science Foundation of China （31201674 and 31371618）;the National Transgenic Major Program, China （2011ZX08008）

Renin and angiotensinogen expression and functions in growth and apoptosis of human glioblastoma

The expression and function in growth and apoptosis of the renin-angiotensin system(RAS)was evaluated inhuman glioblastoma.Renin and angiotensinogen(AGT)mRNAs and proteins were found by in situ hybridisationand immunohistochemistry in glioblastoma cells.Angiotensinogen was present in glioblastoma cystic fluids.Thus,human glioblastoma cells produce renin and AGT and secrete AGT.Human glioblastoma and glioblastoma cellsexpressed renin,AGT,renin receptor,AT(2)and/or AT(1)mRNAs and proteins determined by RT-PCR and/

Functional expression of opioid receptor-like receptorand its endogenous specific agonist nociceptin/orphanin FQ during mouse embryogenesis

Expression of opioid receptor-like receptor (ORL1)and its endogenous peptide agonist nociceptin/orphaninFo (N/OFQ) during mouse embryogenesis have been investigated. Transcripts of ORL1 and N/OFQ were detected by RT-PCR in mouse brain of day 8 embryo (E8)and the expression continued afterwards. Northern blotanalysis revealed abundant expression of ORL1 at postnatal day 1 (P1) and N/OFQ at E17 and P1 in the brain butnone was detected in other embryonic tissues. The presence of functional ORL1 in mouse embryonic brain wasalso confirmed by specific binding of [3H] N/OFQ (kd=1.3±0.5 nM and Bmax = 72±9 fmol/mg protein) as wellas by N/OFQ-stimulated G protein activation.

Transcriptome and Functional Analysis of Fiber-related Gene Expression in Cotton

Fiber cell initiation is a complex process involving many pathways,including phytohormones and components for transcriptional and posttranscriptional regulation.Here we report expression

Expression and functional identification of the hypothetical adhesin P32 from Mycoplasma genitalium

Mycoplasma genitalium is the main causative agent for non-gonococcal and non-chlamydial urethritis. P32 is the putative surface-exposed membrane protein of M. genitalium and it has substaintial identity in amino acid sequence with adhesin protein P30 from M. pneumoniae. Since M. pneumoniae mutants lacking P30 protein is defective in cytadherence, P32 protein has been proposed to be an essential adhesin implicated in the adherence of M. genitaliurn to host cells. The prokaryotic expression vector pET-30 （ ＋ ）/p32 was constructed in the present study, and the recombinant protein was expressed in E. coli and purified under denaturing condition. As demonstrated by the immuno- blotting analysis, the recombinant protein could react with rabbit antisera against M. genitalium, and adherence inhibition assays were performed with antisera against this recombinant protein. It was demonstrated that P32 protein apperared to be an adhesion protein of M. genitalium, thus providing the experimental basis for better understanding of the pathogenesis of M. genitalium infection and for the development of the related vaccines against the infection.

Soluble Expression and Rapid Quantification of GFP-hepA Fusion Protein in Recombinant Escherichia coli

To establish a rapid quantification method for heparinase I during its production in recombinant Escherichia coli, a translational fusion vector was constructed by fusing the N terminus of heparinase I to the C terminus of a green fluorescent protein mutant （GFPmutl）. As a result, not only was the functional recombinant expression of heparinase I in E. coli accomplished, but also a linear correlation was obtained between the GFP fluorescence intensity and heparinase I activity, allowing enzyme activity to be quantified rapidly during the fermentation.

Molecular Therapy and Prevention of Liver Diseases

Molecular analyses have become an integral part of biomedical research as well as clinical medicine. The definition of the genetic basis of many human diseases has led to a better understanding of their pathogenesis and has in addition offered new perspectives for their diagnosis, therapy and prevention. Genetically, human diseases can be classified as hereditary monogenic, acquired monogenic and polygenic diseases. Based on this classification, gene therapy is based on six concepts: (1) gene repair, (2) gene substitution, (3) cell therapy, (4) block of gene expression or function, (5) DNA vaccination and (6) gene augmentation. While major advances have been made in all areas of gene therapy during the last years, various delivery, targeting and safety issues need to be addressed before these strategies will enter clinical practice. Nevertheless, gene therapy will eventually become part of the management of patients with various liver diseases, complementing or replacing existing therapeutic and preventive strategies.

Cloning and Characterization of a Lycium chinense Carotenoid Isomerase Gene Enhancing Carotenoid Accumulation in Transgenic Tobacco

Carotenoid isomerase （CRTISO）is a key enzyme that catalyzes the conversion of cis-lycopene to all- trans tycopene. In this study, we isolated and characterized the CRTISO gene from Lycium chinense （LcCRTISO） for the first time. The open reading flame of LcCRTISO was 1 815 bp encoding a protein of 604 amino acids with a molecular mass of 66.24 kDa. Amino acid sequence analysis revealed that the LcCRTISO had a high level of simi- larity to other CRTISO. Phylogenetic analysis displayed that LcCRTISO kept a closer relationship with the CRTISO of plants than with those of other species. Semi-quantitative PCR analysis indicated that LcCRTISO gene was expressed in all tissues tested with the highest expression in maturing fruits. The overexpression of LcCRTISO gene in transgenic tobacco resulted in an increase of total carotenoids in the leaves with [3-carotene and lutein being the predominants. The results obtained here clearly suggested that the LcCRTISO gene was a promising candidate for carotenoid production.

Research progress of microRNA in prevention and treatment of osteonecrosis of the femoral head

In the 1980s,Feinbaum et al.first discovered miRNA in the development of worms and termed it as Lin-4.With the development of microbial science and continuous improvement in the gene sequencing technology,miRNA has been gradually gaining attention for intensive research in clinical medicine.Among these,one group of miRNAs has been confirmed to play fundamental roles in gene regulation in various orthopedics diseases,such as bone tumors,osteoarthritis,and rheumatoid arthritis.The study of miRNAs in the osteonecrosis of the femoral head(ONFH)can improve the understanding of the pathogenesis of the disease.ONFH is an orthopedic disease that is the primary cause of disrupted blood supply to the femoral head and the main symptoms of bone and muscle dysfunction.Recent studies showed that miRNA played a major role in the regulation of the microcirculation of ONFH,damage and repair of blood vessels,local microcirculation dysfunction caused by other diseases,and apoptosis of bone cells.In this study,recent related research results of miRNA and ONFH were analyzed and summarized,and the prospective in the prevention and treatment of the disease was also discussed.

Semi-tensor product of matrices approach to reachability of finite automata with application to language recognition

This paper investigates the transition function and the reachability conditions of finite automata by using a semitensor product of matrices, which is a new powerful matrix analysis tool. The states and input symbols are first expressed in vector forms, then the transition function is described in an algebraic form. Using this algebraic representation, a sufficient and necessary condition of the reachability of any two states is proposed, based on which an algorithm is developed for discovering all the paths from one state to another. Furthermore, a mechanism is established to recognize the language acceptable by a finite automaton. Finally, illustrative examples show that the results/algorithms presented in this paper are suitable for both deterministic finite automata （DFA） and nondeterministic finite automata （NFA）.

Molecular Cloning and Expression of Squalene Epoxidase from a Medicinal Plant, Bupleurum chinense

Objective In plant, squalene epoxidase （SE） catalyzes the first oxygenation step in the biosynthetic pathway of triterpenoid and phytosterol, representing one of the rate-limiting enzymes in this pathway. Bupleurum chinense is an important medicinal herb with its major active constituents such as triterpenoid saponins and saikosaponins. In order to obtain the series of enzymatic genes involved in saikosaponin biosynthesis, a cDNA of SE, designated BcSEI, was cloned from B. chinense. Methods The BcSEI gene was cloned by homology-based PCR and 5＇/3＇ RACE methods from the adventitious roots of B. chinense. The physical and chemical parameters of BcSE1 protein were predicted by protparam. In order to discover hints in amino acid sequences on the dominant functions in the biosynthesis of saponin or phytosterol, sequences of SE from other plants were downloaded from NCBI for sequences alignment and phylogenetic analysis. BcSEI was cloned into a yeast mutant KLNI （MATa, ergl.＇：URA3, leu2, ura3, and trpl） to verify the enzyme activity of BcSE1. Additionally, the tissue-specific expression and methyl jasmonate （MeJA） inducibility of BcSEI were investigated using quantitative real-time PCR. Results The predicted protein of BcSE1 is highly similar to SEs from other plants sharing amino acid sequence identities of up to 88%. The BcSEI can functionally complement with yeast SE gene （ERGI） when expressed in the KLNI mutant （MATa, ergl：：URA3, leu2, ura3, and trpl）. Using as controls with ^-amyrin synthase （G-AS） which is presumed to catalyze the first committed step in saikosaponin biosynthesis and a cycloartenol synthase （CAS） relating to the phytosterol biosynthesis, the transcript of BcSE1 was significantly elevated by MeJA in adventitious roots of B. chinenseand the transcript of BcSElwas most abundant in the fruits and flowers of plants, followed by that in the leaves and roots, and least in stems. Conclusion It is the first time to illustrate the molecular information of SE in B. chinense and to clone the full-length SEgene in plants of genus Bupleurum L.

Assembly of yield heterosis of an elite rice hybrid is promising by manipulating dominant quantitative trait loci

In the past,rice hybrids with strong heterosis have been obtained empirically,by developing and testing thousands of combinations.Here,we aimed to determine whether heterosis of an elite hybrid could be achieved by manipulating major quantitative trait loci.We used 202 chromosome segment substitution lines from the elite hybrid Shanyou 63 to evaluate single segment heterosis(SSH)of yield per plant and identify heterotic loci.All nine detected heteroticloci acted in a dominant fashion,and no SSH exhibited overdominance.Functional alleles of key yield-related genes Ghd7,Ghd7.1,Hd1,and GS3 were dispersed in both parents.No functional alleles of three investigated genes were expressed at higher levels in the hybrids than in the more desirable parents.A hybrid pyramiding eight heterotic loci in the female parent Zhenshan 97 background had a comparable yield to Shanyou 63 and much higher yield than Zhenshan 97.Five hybrids pyramiding eight or nine heterotic loci in the combined parental genome background showed similar yield performance to that of Shanyou 63.These results suggest that dominance underlying functional complementation is an important contributor to yield heterosis and that heterosis assembly might be successfully promised by manipulating several major dominant heterotic loci.

A Numerical Algorithm for Arbitrary Real-Order Hankel Transform

The Hankel transform is widely used to solve various engineering and physics problems,such as the representation of electromagnetic field components in the medium,the representation of dynamic stress intensity factors,vibration of axisymmetric infinite membrane and displacement intensity factors which all involve this type of integration.However,traditional numerical integration algorithms cannot be used due to the high oscillation characteristics of the Bessel function,so it is particularly important to propose a high precision and efficient numerical algorithm for calculating the integral of high oscillation.In this paper,the improved Gaver-Stehfest(G-S)inverse Laplace transform method for arbitrary real-order Bessel function integration is presented by using the asymptotic characteristics of the Bessel function and the accumulation of integration,and the optimized G-S coefficients are given.The effectiveness of the algorithm is verified by numerical examples.Compared with the linear transformation accelerated convergence algorithm,it shows that the G-S inverse Laplace transform method is suitable for arbitrary real order Hankel transform,and the time consumption is relatively stable and short,which provides a reliable calculation method for the study of electromagnetic mechanics,wave propagation,and fracture dynamics.