[Hanfeng Pre-reservoir Commissioning Time Variation Feature of the Hydrology and Water Quality in Three Gorges Reservoir].

Hanfeng Pre-reservoir is very rare in the world which is specially designed to reduce the impact of Fluctuating Zone, and it is formed in Hanfeng Lake of Three Gorges reservoir. The Hanfeng Pre-reservoir has many special hydrological characteristics and ecological environment features based on its unique "pre-reservoir" control mode, the wide seasonal wetland of Fluctuating Zone, the huge life pollution and agricultural pollution, and the pressure of huge city and excessive population. HanFeng Lake has a variety of morphological features such as lakes, rivers, and other backwater bay, for the effect of water level regulation in Three Gorges, since the successful commissioning of the Hanfeng Lake pre-dam system in 2015.

[Hanfeng Pre-dam Commissioning Eutrophication Status and Control Evaluation in Three Gorges Reservoir].

To reduce the impact of Fluctuating Zone, the Three Gorges Reservoir pre-dam is rare in the world which is specially designed and is the largest artificial lake body in China. The ecological benefits of landscape, farmland and lake and the social benefits of livable city have been significantly enhanced since the successful commissioning of the Hanfeng Lake pre-dam system. The paper proposed the application of layered hydrology and water quality monitoring for analysis of Tributary runoff and lake body section in the pre-dam commissioning in the whole year, and a total of 17 measured indicators inlucding hydrological parameters such as , etc, physical parameters such as , pH, SD, DO, TSS etc.

[Comparison of the electrophysiological features between the rhythmic cells of the aortic vestibule and the sinoatrial node in the rabbit].

The purpose of this study was to clarify the characteristics of the pacemaker cells in the left ventricular outflow tract (aortic vestibule) and compare them with those of the cells in the sinoatrial node (SAN). By using conventional intracellular microelectrode technique to record their action potentials, some ionic channel blockers were used to observe their electrophysiological effects on the two types of pacemaker cells in the rabbit, especially on the ionic movement during phase 0 and phase 4. The results obtained are as follows.

[Detection of 2'-O-ribose Methylation Sites on Rice 25 S rRNA].

Ribose methylation is a widespread type of nucleotide modification in rRNA. In order to map the methylation sites of rice 25 S rRNA, a series of primers complementary to both yeast 28 S and rice 25 S rRNA simultaneously were synthesized. Primer extensions at different dNTP concentrations were carried out to detect the methylation sites of both yeast and rice rRNAs.

Studies on interaction between hTNF-alpha and its two receptors with expressed hsTR55-preS1/hsTR75-preS1 fusion soluble receptors.

The gene encoding the N-terminal 2-50 amino acids of HBsAg-preS1 was amplified by PCR and fused to the 3'-end of two human soluble TNF receptor genes to form the hsTR55-preS1/hsTR75-preS1 fusion genes. The recombinant bicistronic expression vectors were further constructed, which contained one of the human soluble TNF receptor fusion genes and the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), followed by the neomycin phosphotransferases as the selectable marker. BHK-21 cells transfected with those vectors by electroporation were selected with G-418, and the positive colonies expressing the protein of interest were obtained.

[Identification of a snoRNA47 gene cluster in Oryza sativa].

Small nucleolar RNAs (snoRNAs) are required for ribose 2'-O-methylation of eukaryotic ribosomal RNA. By researching in international rice genome databases, a snoRNA gene cluster, consisting of three box C/D snoRNA gene candidates, was found on chromosome 6. All of snoRNA coding sequences in this cluster exhibited the characteristic structure of box C/D antisense snoRNA.

Analysis of C-terminal Amidating Structure and Determination of Amidating Enzyme Activity by Using Capillary Electrophoresis.

Based on the difference in charge and hydrophobicity between amidating moiety and the carboxyl group in a given buffer solution, a new method to analyze the C-terminal amidating structure and the amidating enzyme activity by using capillary electrophoresis was established.

Selection with SELEX Method of Small RNA Molecules Specifically Binding to Starch.

A 73-base DNA library with T7 promoter was chemically synthesized according to the sequence specified by the 2.5S RNA component of glycogen branching holoenzyme (EC 2.4.

Efficient Secretion of Proteins Expressed from Insect Cells Directed by PAM Signal Peptide.

Signal and leading peptide sequences of rat PAM was inserted into the baculovirus transfer vector, and secretion expression plasmids pBACPAG2 and pBacPAI for the fusion gene PABC-hGRF and PABC-IGF-I were constructed, respectively. By cotransfection with linear genomic DNA of modified Autographa californica nuclear polyhedrosis virus (BacPAK6) and homologous recombination, the recombinant AcNPV, BacPAG and BacPAI, were obtained and identified. Fusion proteins PABC-hGRF and PABC-IGF-I were secreted efficiently from Sf21 cells infected with BacPAG and BacPAI, respectively, and those fusion protein could be purified efficiently by IgG affinity column.

Expression in BHK Cells of Two Human TNF Receptors and Their Interaction with hTNF-alpha.

Two human tumor necrosis factor receptor genes were cloned into eukaryotic expression vectors pcDNA3, pDR2 and pXJ-41, respectively. Those vectors were transfected into BHK-21 cells and the expression of hTNFRs was identified and demonstrated by binding of (125)I hTNF-alpha and Scatchard analysis. The expression of two hTNFRs in BHK cells at the RNA transcriptional and the protein translation levels were also demonstrated by RT-PCR and indirect ELISA.

A Discriminator Base in tDNA(Trp).

The discriminator base G73 was one of the major identity elements of tRNA(Trp). Aminoacylation assay and CD spectrum of tDNA(Trp)-NCCrA demonstrated that under the same pH, the change of discriminator base affected tRNA conformation at the 3' terminus while tDNA(Trp) with the same discriminator base exhibited different conformation and aminoacylation activity under various pH conditions. Our results showed that it was the conformation of tRNA, but not the base itself, that determined the accurate recognition between tRNA and its cognate synthetase.

Isolation and Identification of Membrane Proteins Binding to Transfer RNA.

Crude extract of total membrane proteins of yeast was obtained by a method of phase separation in Triton X-114. The membrane extract was applied to the affinity column on which yeast total tRNAs were covalently coupled to hydrazinyl Sepharose 4B. By eluting with increasing concentration gradient of (NH(4))(2)SO(4), eluted proteins were found between concentrations of (NH(4))(2)SO(4) 0.

Construction and Expression of hTNFalpha-hTGF3 Fused Gene.

Human tumor necrosis factor was fused with hTGF3, the third loop region of hTGFalpha specifically binding to EGF receptor, through a flexible linker by recombinant DNA techniques. The constructed plasmid containing hTNFalpha-hTGF3 fused gene, pSB92-TLT, was expressed under the control of P(L) promoter and after the induction at 42 degrees it gave a high level of expression of the fused gene, ranging in 50%-60% of total bacterial protein. 95% of the expressed product was inclusion body, and only 5% was soluble.

SELEX Screening and Characterization of Small RNA Molecules That Specifically Bind the Reactive Blue Dye.

A 73-base single-stranded DNA library with a T7 promoter was chemically synthesized, in which 20 consecutive bases were completely randomized. After PCR amplification and T7 RNA polymerase transcription of the DNA random library, the in vitro transcribed RNA random library was subjected to 8 successive rounds of reactive blue dye column selection by SELEX method, resulting in a sharp increase of the percentage of the transcribed RNA pool that could bind reactive blue dye, from less than 0.03% in the first round pool to 22.

Properties of Recombinant Aeromonas punctata Prolyl Endopeptidase.

The properties of recombinant Aeromonas punctata prolyl endopeptidase(apPEP) were studied using specific substrate and peptides. Results show that the optimum catalytic temperature and pH was 34 degrees and 8.4, the stability of the apPEP was in the range of 4-32 degrees and pH 6.

Overexpressed Human TNF Receptor-75 Can Independently Mediate hTNFalpha-induced Cytotoxicity in BHK-21 Cells.

A recombinant bicistronic expression vector was constructed containing human TNF receptor-75 gene and encephalomyocarditis virus(EMCV)internal ribosome entry site(IRES), followed by a neomycin phosphotransferase gene as selectable marker. BHK-21 cells were transfected with this vector using electroporation method and positive clones overexpressing the genes of interest were obtained after selection with G-418. The expression of two hTNFRs in those cells at RNA transcriptional and the protein translation levels had been proved by RT-PCR and indirect ELISA.

Immobilized Tryptophanyl-tRNA Synthetase from Bacillus subtilis.

In order to investigate the recognition mechanism and the relationship between structure and function of tRNA(Trp) with tryptophanyl-tRNA synthetase (TrpRS), TrpRS from Bacillus subtilis was purified and immobilized on CNBr-activated Sepharose 4B. Protein recovery and activity recovery of the immobilization were 95.5% and 31.

Mitochondrial tRNA(Trp)s Imply a Eubacterial Origin.

Three mitochondrial tRNA(Trp)s genes from Oryza sativa, Homo sapiens and Saccharomyces serevisea were constructed. In vitro transcripts of these mitochondrial tRNA(Trp)s genes were able to be tryptophanylated by Bacillus subtilis tryptophanyl-tRNA synthetase (TrpRS), but could not be catalyzed by TrpRS from rat liver. Kinetic assay showed that B.

Cloning and Expression of the cDNA Coding for Rat Peptidylglycine alpha-Amidating Monooxygenase.

The dDNA of genes encoding rat peptidylglycine alpha-aminating monooxygenase(rPAM) were cloned and their expression in E. coli studied. Three DNA fragments were isolated from the rat brain cDNA library using the methods of plaque hybridization and PCR.