[Research development of the interaction of small molecules with nucleic acids].
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Animals↗
Biomedical subjects
J Liu
Publications and source records attributed to J Liu.
[Studies on the arbutin biosynthesis by hairy root of Panax ginseng C.A. Mayer].
OBJECTIVE: To study basic conditions for biosynthesis arbutin by the hairy root of Panax ginseng. METHOD: Based on the content of arbutin and the biotransformation rate of hydroquione, these conditions such as the culture stage, the lasting time and the concentration of hydroquione fitting for biosynthesis were found. RESULT: After cultured for 22 days, transferred in fresh B5 medium having 2 mmol.L-1 hydroquione, the hairy root biotransformated hydroquione into arbutin in 24 hours, the bioconversion rate was 89.0% and the content of arbutin in dry root was 13.0%. CONCLUSION: Arbutin, was which never found in the genus of Panax ginseng, can be biosynthesized by the hairy root of Panax ginseng.
Arbutin↗
Expression and deletion analysis of EcoRII endonuclease and methylase gene.
OBJECTIVE: To clone complete EcoRII restriction endonuclease gene (ecoRIIR) and methyltransferase gene (ecoRIIM) in one vector and to analyze the coordinating expression of this whole R-M system. METHODS: Unidirectional deletion subclones were constructed with ExoIII. ecoRIIR/ M genes were preliminarily located in the cloned fragment according to the enzyme activities of subclones. Exact deletion sites were determined by sequencing, and transcriptional start sites were determined by S1 mapping. RESULTS: The DNA fragment which was cloned into pBluescript SK + contained intact ecoRIIR gene and ecoRIIM gene, and two transcriptional start sites of ecoRIIR gene were determined. 132bp to 458bp from 3'end of ecoRIIR gene are indispensable to enzyme activities and deletion of 202bp from 3' end of ecoRIIM gene made enzyme lose the capability in DNA protection to resist specific cut with EcoRII endonuclease (EcoRII. R). Deletion of the coding and flanking sequences of one gene did not affect the expression of the other gene, and the recombinants only containing ecoRIIR gene appeared to be lethal to dcm+ host. CONCLUSION: ecoRIIM gene linking closely to ecoRIIR gene is very important for the existence of the R-M system in process of evolution, but the key to control EcoRII R-M order may not exist in transcriptional level .
Base Sequence↗
[Determination of lead and arsenic in copper aspirinate by graphite furnace atomic absorption spectrometry].
A graphite furnace atomic absorption spectrometric method has been established to determine trace lead and arsenic in copper aspirinate, a new anti-inflammatory and anti-thrombotic agent. The sample is pretreated by ashing at 530 degrees C in the presence of Ni (NO3)2, followed by dissolution with HNO3. The method is simple and has a low detection limit. The relative standard deviation is 11%-15%, the recovery is 92%-112% for lead and 86%-119% for arsenic.
Anti-Inflammatory Agents, Non-Steroidal↗
[Spectroscopy study on the photochromism of Schiff base N-salicylidene-1-hexadecylamine].
The photochromism of N-Salicylidene-1-Hexadecylamine was studied by time-dependent fluorescence spectra, UV-Vis absorption spectra and theoretical chemistry calculation method. The photo-product was determined to be the keto form. The dynamics of the photochromism of the title compound was studied by the time course technique.
English Abstract↗
[Study on the deteriorating course of fresh milk by laser-induced fluorescence spectra].
Along with the development of living standard, people's demand for food quality and food hygiene also rises. People demand food not only with rich nutrition, inexpensive price, but also with safety. So food hygiene test is paid common attention of society. Milk is a nourishing food and is loved by people. Sour milk goods from milk is also in great demand. But nourishing foods are good for growing many microbes. Fresh milk and sour milk are easy contaminated by microbes and go bad. Laser-induced fluorescence (LIF) technology is an important part of modern optics. It is broadly applied in biomedicine, diagnostics, test of food hygiene, environment protecting, owing to its high sensitivity, high speed, automation, untouched testing. In this paper, we attempted to LIF technology to test milk food quality. We used the third harmonics pulsed Nd:YAG laser (355 nm) as the exciting source, and a multi-track spectrometer as the detector and measured the intensities of apply LIF of fresh milk and sour milk during their deteriorating course. Test system and test method are introduced, fluorescence spectra of deteriorating course are also attached. The test result makes clear that there are close connection between deteriorating course and fluorescence spectra.
Animals↗
[The relationship between clinical prognosis and classification of the isoforms of fuse gene in acute promyelocytic leukemia].
To check the anti-apoptosis gene bcl-2 protein/mRNA expression in 28 acute promyelocytic leukemia (APL), by immunohistochemistry and in situ hybridization, and to analyse the relationship among the bcl-2 gene expressions, clinical prognosis and classification of the isoforms of fuse gene in APL. The result shows the two types of isoforms of PML-RAR alpha have no significance in bcl-2 protein/mRNA expression, indexes of clinical prognosis. This shows that the classification of PML-RAR alpha isoform has no influence on clinical prognosis.
Adolescent↗
DNA polymerase beta: contributions of template-positioning and dNTP triphosphate-binding residues to catalysis and fidelity.
The specific catalytic roles of two groups of DNA polymerase beta active site residues identified from crystal structures were investigated: residues possibly involved in DNA template positioning (Lys280, Asn294, and Glu295) and residues possibly involved in binding the triphosphate moiety of the incoming dNTP (Arg149, Ser180, Arg183, and Ser188). Eight site-specific mutants were constructed: K280A, N294A, N294Q, E295A, R149A, S180A, R183A, and S188A. Two-dimensional NMR analysis was employed to show that the global conformation of the mutants has not been perturbed significantly. Pre-steady-state kinetic analyses with single-nucleotide gapped DNA substrates were then performed to obtain the rate of catalysis at saturating dNTP (k(pol)), the apparent dissociation constant for dNTP (K(d)), catalytic efficiency k(pol)/K(d), and fidelity. Of the three template-positioning residues, Asn294 and Glu295 (but not Lys280) contribute significantly to k(pol). Taken together with other data, the results suggest that these two residues help to stabilize the transition state during catalysis even though they interact with the DNA template backbone rather than directly with the incoming dNTP or the opposite base on the template. Furthermore, the fidelity increases by up to 19-fold for N294Q due to differential k(pol) effects between correct and incorrect nucleotides. Of the four potential triphosphate-binding residues, Ser180 and Arg183 contribute significantly to k(pol) while the effects of R149A are relatively small and are primarily on K(d), and Ser188 appears to play a minimal role in the catalysis by Pol beta. These results identify several residues important for catalysis and quantitate the contributions of each of those residues. The functional data are discussed in relation to the prediction on the basis of available crystal structures.
Animals↗
Magnetic-field-induced structural transitions in a ferrofluid emulsion.
A ferrofluid emulsion, subjected to a slowly increasing magnetic field, exhibits a complicated structural behavior: a gas of Brownian particles changes to columnar solid structures due to induced dipole interaction. Two transition (intermediate) structural regimes are observed: (i) randomly distributed chains and particles and (ii) distinct thin columns and randomly distributed chains and particles. Three structural transition magnetic fields are found, one marking each structural transition, from the initial to the final structural regime. A structural diagram of the structural transition magnetic fields, H(C), versus particle volume fractions, straight phi, is constructed experimentally. Theoretical models of scaling calculations, based upon the dominant magnetic interaction in each structural regime, give the three structural transition magnetic-field relations as H(C1) proportional to straight phi(-1/2), H(C2) proportional to straight phi(-1/4), and H(C3) proportional to (straight phi(gamma)/G2)exp(piG/straight phi((gamma/2))), where gamma=0.39 and G=0.29 for our sample. The final end shape of columns and the relative position between columns show that the end-end repulsion between chains is important in the structural formation.
Journal Article↗
Regulation of apoptosis and cell cycle progression by MCL1. Differential role of proliferating cell nuclear antigen.
MCL1 (ML1 myeloid cell leukemia 1), a Bcl-2 (B- cell lymphoma-leukemia 2) homologue, is known to function as an anti-apoptotic protein. Here we show in vitro and in vivo that MCL1 interacts with the cell cycle regulator, proliferating cell nuclear antigen (PCNA). This finding prompted us to investigate whether MCL1, in addition to its anti-apoptotic function, has an effect on cell cycle progression. A bromodeoxyuridine uptake assay showed that the overexpression of MCL1 significantly inhibited the cell cycle progression through the S-phase. The S-phase of the cell cycle is also known to be regulated by PCNA. A mutant of MCL1 that lacks PCNA binding (MCL1(Delta)(4A)) could not inhibit cell cycle progression as effectively as wild type MCL1. In contrast, MCL1(Delta)(4A) retained its anti-apoptotic function in HeLa cells when challenged by Etoposide. In addition, the intracellular localization of MCL1(Delta)(4A) was identical to that of wild type MCL1. An in vitro pull-down assay suggested that MCL1 is the only Bcl-2 family protein to interact with PCNA. In fact, MCL1, not other Bcl-2 family proteins, contained the PCNA-binding motif described previously. Taken together, MCL1 is a regulator of both apoptosis and cell cycle progression, and the cell cycle regulatory function of MCL1 is mediated through its interaction with PCNA.
Amino Acid Sequence↗
Determination of PyPuPu (PyPuPy) intermolecular triple-stranded DNA by capillary electrophoresis.
The PyPuPu and PyPuPy intermolecular triple-stranded DNA (tsDNA) can be determined more easily by capillary electrophoresis (CE) than by traditional methods. The tsDNA and its component compounds can be well separated by using a sieving matrix of 1.0% hydroxypropylmethylcellulose (HPMC) containing 2.5 mM magnesium ions. Such factors as buffer pH, the concentration of triplex-forming oligonucleotide (TFO), temperature, and the concentration of magnesium cation in the formation and stabilization of triple-stranded helices have been studied with capillary electrophoresis. The triplex cannot be formed when the buffer pH is lower than 4.0. When the concentration of TFO is four times higher than that of dsDNA, all of the dsDNA molecules can be associated. The limit of capillary electrophoresis detection with good reproducibility is 0.5-1 nM (S/N = 3). The CE analysis of short tsDNA takes only 40 min, whereas gel electrophoresis needs at least 5 h.
Antineoplastic Agents↗
Tyrosine phosphorylation of cortactin is required for H2O2-mediated injury of human endothelial cells.
Injury of endothelial cells induced by reactive oxygen species plays an important role in the development of early stages of vascular diseases such as hypertension and atherosclerosis. Exposure of human umbilical vein endothelial cells to hydrogen peroxide (H(2)O(2)), a common form of reaction oxygen species, triggers a series of intracellular events, including actin cytoskeletal reorganization, cytoplasm shrinkage, membrane blebbing and protein-tyrosine phosphorylation. The effect of H(2)O(2) on endothelial cells is dramatically enhanced when a survival pathway involving extracellular signal-regulated kinase is blocked by PD098059. In contrast, the injury of endothelial cells mediated by H(2)O(2) is inhibited by PP2, a selective specific inhibitor for protein-tyrosine kinase Src. Cortactin, a filamentous actin (F-actin)-associated protein, becomes phosphorylated at tyrosine residues upon stimulation by H(2)O(2) in a manner dependent on the activity of Src. The level of tyrosine phosphorylation of cortactin is correlated with the formation of membrane blebs. Overexpression of wild-type cortactin tagged with green fluorescent protein in endothelial cells via a retroviral vector substantiates the H(2)O(2)-induced morphological changes, whereas overexpression of a green fluorescent protein-cortactin mutant deficient in tyrosine phosphorylation renders endothelial cells resistant to H(2)O(2). The functional role of cortactin in H(2)O(2)-mediated shape changes was also evaluated in NIH 3T3 cells. Stable 3T3 transfectants expressing wild-type cortactin in the presence of either H(2)O(2)/PD098059 or H(2)O(2) alone at 200 microm exhibited a dramatic shape change characterized by rounding up or aggregation. However, the similar changes were not detected with cells overexpressing a cortactin mutant deficient in tyrosine phosphorylation. These data demonstrate an important role of the Src/cortactin-dependent actin reorganization in the injury of endothelial cells mediated by reactive oxygen species.
CSK Tyrosine-Protein Kinase↗
Health and war in Congo-Brazzaville.
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Congo↗
A deoxyinosine specific endonuclease from hyperthermophile, Archaeoglobus fulgidus: a homolog of Escherichia coli endonuclease V.
Deoxyadenosine undergoes spontaneous deamination to deoxyinosine in DNA. Based on amino acids sequence homology, putative homologs of endonuclease V were identified in several organisms including archaebacteria, eubacteria as well as eukaryotes. The translated amino acid sequence of the Archaeoglobus fulgidus nfi gene shows 39% identity and 55% similarity to the E. coli nfi gene. A. fulgidus endonuclease V was cloned and expressed in E. coli as a C-terminal hexa-histidine fusion protein. The C-terminal fusion protein was purified to apparent homogeneity by a combination of Ni(++) affinity and MonoS cation exchange liquid chromatography. The purified C-terminal fusion protein has a molecular weight of about 25kDa and showed endonuclease activity towards DNA containing deoxyinosine. A. fulgidus endonuclease V has an absolute requirement for Mg(2+) and an optimum reaction temperature at 85 degrees C. However, in contrast to E. coli endonuclease V, which has a wide substrate spectrum, endonuclease V from A. fulgidus recognized only deoxyinosine. These data suggest that the deoxyinosine cleavage activity is a primordial activity of endonuclease V and that multiple enzymatic activities of E. coli endonuclease V were acquired later during evolution.
Amino Acid Sequence↗
In vitro selection of an RNA sequence that interacts with high affinity with thymidylate synthase.
Previous studies have shown that the repressive effect of thymidylate synthase (TS) mRNA translation is mediated by direct binding of TS itself to two cis-acting elements on its cognate mRNA. To identify the optimal RNA nucleotides that interact with TS, we in vitro synthesized a completely degenerate, linear RNA pool of 25 nt and employed in vitro selection to isolate high affinity RNA ligands that bind human TS protein. After 10 rounds of selection and amplification, a single RNA molecule was selected that bound TS protein with nearly 20-fold greater affinity than native, wild-type TS RNA sequences. Secondary structure analysis of this RNA sequence predicted it to possess a stem-loop structure. Deletion and/or modification of the UGU loop element within the RNA sequence decreased binding to TS by up to 1000-fold. In vivo transfection experiments revealed that the presence of the selected RNA sequence resulted in a significant increase in the expression of a heterologous luciferase reporter construct in human colon cancer H630 and TS-overexpressing HCT-C:His-TS+ cells, but not in HCT-C18 cells expressing a functionally inactive TS. In addition, the presence of this element in H630 cells leads to induced expression of TS protein. An immunoprecipitation method using RT-PCR confirmed a direct interaction between human TS protein and the selected RNA sequence in transfected human cancer H630 cells. This study identified a novel RNA sequence from a degenerate RNA library that specifically interacts with TS.
Base Sequence↗
Novel ferrocenyl polyene derivatives and their binding to unmodified cyclodextrins
Two new ferrocene derivatives, 7-ferrocenyl-2,4,6-heptatrienal (1) and 7-ferrocenyl-2,4,6-heptatrienol (2), were synthesized and characterized. These two compounds possess a rigid triene chain conjugated to one of the cyclopentadienyl rings of the ferrocene residue, and as a result, they exhibit very stiff structures. The electronic absorption and electrochemical properties of these compounds were utilized to investigate their host-guest binding interactions with the receptors alpha-cyclodextrin (alpha-CD) and beta-cyclodextrin (beta-CD) in aqueous solution. From electronic absorption measurements binding constants in the range 790-12900 M(-)(1) were obtained; beta-CD formed more stable complexes than alpha-CD with both guests. Electrochemical measurements suggest some degree of site selectivity in the complexation processes, with beta-CD binding preferentially to the ferrocene moiety while alpha-CD interacts with the unsaturated chain.
Journal Article↗
Synthesis of a substance P antagonist with a somatostatin scaffold: factors affecting agonism/antagonism at GPCRs and the role of pseudosymmetry.
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Animals↗
Headspace solid-phase microextraction and gas chromatographic-mass spectrometric screening for volatile hydrocarbons in blood.
Optimization for headspace solid-phase microextraction (SPME) was studied with a view to performing gas chromatographic-mass spectrometric (GC-MS) screening of volatile hydrocarbons (VHCs) in blood. Twenty hydrocarbons comprising aliphatic hydrocarbons ranging from n-hexane to n-tridecane, and aromatic hydrocarbons ranging from benzene to trimethylbenzenes were used in this study. This method can be used for examining a burned body to ascertain whether the victim had been alive or not when the burning incident took place. n-Hexane, n-heptane and benzene, the main indicators of gasoline components, were found as detectable peaks through the use of cryogenic oven trapping upon SPME injection into a GC-MS instrument. The optimal screening procedure was performed as follows. The analytes in the headspace of 0.2 g of blood mixed with 0.8 ml of water plus 0.2 microg of toluene-d8 at -5 degrees C were adsorbed to a 100-microm polydimethylsiloxane (PDMS) fiber for 30 min, and measured using the full-mass-scanning GC-MS method. The lower detection limits of all the compounds were 0.01 microg per 1 g of blood. Linearities (r2) within the range 0.01 to 4 microg per 1 g of blood were only obtained for the aromatic hydrocarbons at between 0.9638 (pseudocumene) and 0.9994 (toluene), but not for aliphatic hydrocarbons at between 0.9392 (n-tridecane) and 0.9935 (n-hexane). The coefficients of variation at 0.2 microg/g were less than 8.6% (n-undecane). In conclusion, this method is feasible for the screening of volatile hydrocarbons from blood in forensic medicine.
Burns↗