Co-administration of Interleukin-2 Enhances Cellular and Humoral Immune Responses to HIV Vaccine DNA Prime／MVA Boost Regime

Interleukine-2 (IL-2) is a growth factor for antigen-stimulated T lymphocytes and is responsible for T-cell clonal expansion after antigen recognition. It has been demonstrated that DNA vaccine-elicited immune responses in mice could be augmented substantially by using either an IL-2 protein or a plasmid expressing IL-2. Twenty mice, divided into four experimental groups, were immunized with: (1) sham plasmid; (2) HIV-1 DNA vaccine alone; (3) HIV-1 DNA vaccine and IL-2 protein; or (4) HIV-1 DNA vaccine and IL-2 plasmid, separately. All the groups were immunized 3 times at a 2-week interval. Fourteen days after the last DNA vaccine injection, recombinant MVA was injected into all the mice except those in group 1. ELISA and ELISPOT were employed to investigate the effect of IL-2 on DNA vaccine immune responses. The obtained results strongly indicate that the efficacy of HIV vaccine can be enhanced by co-administration of a plasmid encoding IL-2.     

REGULATION OF GLUCOCORTICOID RECEPTOR BY GLUCOCORTICOIDS(Ⅱ) ——THE STUDIES ON RAT LIVER, BRAIN AND SPLEEN

The concentration of glucocorticoids (GC) in plasma was maintained at stress level, 20--40μg/dl, for 3 days by subcutaneous injection of hydrocortisone (F) in polyvinyl alcohol (PVA) into rats, and the specific binding of [~3H]Dexamethasone (Dex) in liver, spleen and brain was determined before and after injection. The binding capacity of glucocorticoid receptor (GR) in liver and spleen was decreased significantly 1 h after injection and maintained at low level for several days after the concentration of GC in plasma had returned to the normal level. The K_d was not altered. The changes of GR in brain was not significant. Thus it may be concluded that GC can down-regulate GR in rats, but with different characteristics in various target organs.     

Endonuclease-based Method for Detecting the Sequence Specific DNA Binding Protein on Double-stranded DNA Microarray

The double-stranded DNA (dsDNA) probe contains two different protein binding sites. One is for DNA- binding proteins to be detected and the other is for a DNA restriction enzyme. The two sites were arranged together with no base interval. The working principle of the capturing dsDNA probe is described as follows: the capturing probe can be cut with the DNA restriction enzyme (such as EcoR I) to cause a sticky terminal, if the probe is not bound with a target protein, and the sticky terminal can be extended and labeled with Cy3-dUTP by DNA polymerase. When the probe is bound with a target protein, the probe is not capable to be cut by the restriction enzyme because of space obstruction. The amount of the target DNA binding proteins can be measured according to the variations of fluorescent signals of the corresponding probes.     

Specific hydrolysis of DNA by Ce~(4+) -ODN hybrid

The cerium complex bound to the derivative of oligoDNA has been synthesized successfully that can hydrolyze DNA with sequence-specificity. The synthesized derivative of oligoDNA, 5'-EDTA-P-10 mers ODN, was demonstrated by HPLC. The fluorescence spectrum of Tb3+ was detected after its interaction with the hybrid of 10-mers ODN and 26-mers ODN and the results show that the artificial endo-enzyme can recognize and combine firmly with the substrate DNA. The electrophorogram shows that the cerium-oligoDNA hybrid can specifically hydrolyze its substrate DNA and the cleavage site of this hydrolysis reaction is also discussed. This artificial nuclease can be widely used in molecular biology and genetic engineering as one kind of endo-enzyme.     

SPONTANEOUS POTENTIALS OF EMBRYONIC EPITHELIUM IN URODELES (CYNOPS ORIENTALIS)

The spontaneous potentials (SPs) of nerveless embryonic epithelium of Cynops orientaliswere observed by conventional intracellular micro-electrode technique. The amplitude and fre-quency of SPs of epithelial cells and the course of initiation and decline were recorded. TheSPs can occur repeatedly at the stages when the embryonic epithelium is able to conduct exci-tation. The SP is very similar to the evoked propagatable potential, but has shorter duration.The amplitudes and frenquencies of SPs differ in the different embryos and in differentepithelial cells in the same embryo. The excitable epithelial cells of the embryo may lose theirconductivity when the SPs come to a stable phase both in amplitude and frequency. And theconductivity can recover again after the subsiding of SPs. The SPs can be eliminated by tetro-dotoxin (TTX), but they are not affected by the treatment of cobalt chloride.     

A Novel Ionic Liquid／water Biphasic System for the Preparation of Oximes

A variety of carbonyl compounds can be converted into oximes efficiently and conveniently in a novel ionic liquid/water bi-phasic system in the presence of sodium bicarbonate at ambient temperature. The ionic liquid 1-butyl-3-methyl imidazolium hexafluorophosphate [bmim]PF6 is immiscible with water or diethyl ether and can be easily recycled for reuse without noticeable droping in activity after separation of the products. The protocol is rapid, the yields are excellent, the method is simple and the ionic liquid can be reused.     

LONG-RANGE CORRELATIONS IN DNA SEQUENCES USING TWO-DIMENSIONAL DNA WALKS

The characterization of long-range correlations and fractal properties of DNA sequences has proved to be a difficult though rewarding task mainly due to the mosaic character of DNA consisting of many patches of various lengths with different nucleotide constitutions. In this paper we investigate statistical correlations among different positions in DNA sequences using the two-dimensional DNA walk. The root-mean-square fluctuation F(l) is described by a power law. The autocorrelation function C(l), which is used to measure the linear dependence and periodicity, exists a power law of C(l) -τμ. We also calculate the mean-square distance ＜R2(l)＞ along the DNA chain, and it may be expressed as ＜R2(l)＞ - l r with 2 ＞γ＞ 1. Our investigations can provide some insights into long-range correlations in DNA sequences.     

Design and Synthesis of Lipids Bearing Nucleosides as Gene Vectors

Delivery of exogenous DNA into cells of various origins plays an important role not only in basic research, but also in clinical application in gene therapy. Great efforts have been made towards the development of viral and nonviral vectors for these purposes1~6. However, at the present stage, none of the vectors can claim to be the panacea. Although most viral vectors can transfer gene efficiently and some of them can be used for gene transfer into tissue cells in vivo, they suffer from immu…     

Optimization of Processing Parameters for Minimizing Warpage of Large Thin-walled Parts in Whole Stages of Injection Molding

This study investigated total warpage of a type of motorcycle seat support made of polypropylene（PP） during the entire process of injection molding and free-cooling after demolding. Finite element modeling（FEM） analysis for injection molding and its associated thermal deformation was carried out in the study. The effects of processing parameters on warpage occurring in different stages were analyzed by Taguchi optimization method. It was found that packing pressure is the major factor that affects warpage in the injection stage, whereas cooling time is the major factor in free-cooling stage. From an overall evaluation, melt temperature affects the total warpage most, followed by cooling time, packing pressure, packing time and mold temperature. The result proved that optimum parameters for minimizing final warpage of the injected parts can be obtained only when the deformation in the entire manufacturing process is addressed in both molding and demolding stages.     

USE OF HUMAN ERYTHROCYTE GHOSTS FOR TRANSFER OF~(125)I-BSA AND~(125)I-DNA INTO ANIMAL CELLS THROUGH CELL FUSION

~(125)I-BSA and human ~(125)I-DNA fragments have been prepared for tests of loading efficiencyby the erythrocyte ghosts, which is found to be 3.7 and 3.0 per cent respectively. The experi-ments are reproduceable. The ~(125)I-DNA fragments trapped in the erythrocyte ghosts aretransferred into DON cells through cell fusion or "microrinjection". The injection efficiencyappraised by autoradiography amounts to 58 per cent.     

Spectral and Electrochemical Investigation of Intercalations of Adrenaline and CT－DNA

A strong interaction between double stranded calf-thymus DNA (ds-DNA) and adrenaline in solution, but no interaction between single stranded calf-thymus DNA (ss-DNA) and adrenaline was observed by the use of UV-visible spectroscopy and voltammetric techniques. It is suggested that the interaction leads to an intercalation of adrenaline molecules into the groove of ds-DNA and the formation of ds-DNA (adrenallne)n complex. The binding site size of the interaction of adrenaline with CT-DNA in nucleotide phosphate [ NP] has been determined as 25. The interaction of different concentration adrenaline with DNA modified GCE shows that the DNA modified GCE can be a good tool to detect lower concentration adrenaline.     

PURIFICATION AND CHARACTERIZATION OF A DNA POLYMERASE-TEMPLATE COMPLEX IN MOUSE ASCITES TUMOR CELLS

A DNA polymerase-template complex found in mouse Erhlich ascites tumor cells waspurified by twice discontinuous gradient centrifugations and agarose gel filtration. Afterpurification, the specific activity of the complex increased about 500-fold. The size of thecomplex was found to be 510,000 daltons. The shape of the complex was globular under electronmieroscope. Its diameter was between 100--110 A. The complex was isolated into two mainenzyme proteins and a homogeneous DNA template by DEAE cellulose chromatography. Themain enzyme had a molecular weight of 300,000 daltons and a sedimentation coefficient of9.5s. The endogenous template DNA showed a single zone in polyacrylamide gel electropho-resis analysis. The sedimentation coefficient was determined to be 3.8s.     

EXPRESSION OF A ZEIN GENE (Z4) INTRODUCED INTO DICOTYLEDONOUS Solanum nigrum

A maize genomic clone containing a zein gene (Z4) is inserted into the T-DNA of the Ti plasmid pTiT37. Agrobacterium tumefaciens strain harboring this modified Ti plasmid is used to infect stem sections of young plants or explants of dicotyledonous Solanum nigrum. Axenic transformed calli active in nopaline synthesis are obtained and transgenic plants are differentiated from them DNA Southern hybridization and RNA dot-hybridization analyses show that the zein gene is really transferred and integrated into the nuclear genome of transformed Solanum nigrum and that the zein gene can be transcribed into mRNA in the transformed calli and shoots. But the presence of the zein protein cannot be detected in either the transformed calli or the transgenic shoots. The results of thte experiments demonstrate that the promoter of a gene from monocotyledonous plants can function normally in transgenic dicots. The possibility of developmentally-regulated expression of the zein gene in transformed dicots is discussed in     

PERIODIC AND NONPERIODIC OSCILLATIONS OF NONLINEAR ATMOSPHERIC WAVES FORCED BY PERIODIC THERMAL FORCING——SYSTEM WITH DISSIPATION

This is a continued paper of [1]. The dissipation is considered here for analysing the mul-tiple equilibria and their instability. It is found that the far super-resonant and sub-resonantequilibria are stable and the near super-resonant equilibrium is unstable. The numerical resultsfor the periodic varying forcing show that the two kinds of stable equilibria in the dissipativesystem can alternatively transform into an asymmetrical oscillation with a period same asthat of the forcing.     

Purification and Characterization of DNA Methylase From HL-60 Cells

A solid leukemia sarcoma has been successfully developed after subcutaneous inoculation of the cultured human promyelocytic leukemia cells (HL-60 cells) into unde mice. The solid leukemia sarcoma is a more plantiful source than the cultured cells for enzymatic study and its growing environment is closer to that of the human body than the cultured cells.We establish an efficient procedure of purifying HL-60 cells DNA methylase which includes: disruption of HL-60 cells by homogenization and sonication, removing the cell fragments and cellular particles by centrifuge and ultracentrifuge (105.000 g); removing endogenous DNA by streptomycin sulfate, salting out by (NH4)2SO4, ion exchange chromatography on DEAE-cellulose (DE-52), gel filtration over Sephadex G-100 column.The DNA methylase from HL-60 cells has been purified 204 fold by this procedure. The purified enzyme shows a single-band on PG-PAGE. A 479-kD molecular weight of this enzyme is measured by PG-PAGE. The enzyme properties of HL-60 DNA methylase     

Selective Sensing Characteristics of Ca Doped BeO Nano-sized Tube toward H20 and NH3

By means of density functional calculations, the structural and electronic properties of chemical modification of pristine and Ca-doped BeO nanotubes were investigated with NH3 and H20 molecules. It was found that the NH3 and H20 molecules can be adsorbed on the Be atom of the tube sidewall with the adsorption energies of about 36.1 and 39.0 kcal/mol, respectively. Density of states analysis shows that the electronic properties of the BeONT are slightly changed after the adsorption processes. Substitution of a Be atom in the tube surface with a Ca atom increases the adsorption energies by about 7.4 and 14.7 kcal/mol for NH3 and H20, respectively. Unlike the pristine tube, the electronic properties of Ca-doped BeONT are sensitive to NH3 and H20 molecules. Also, the Ca-doped tube is much more sensitive to H20 molecule than NH3 one.     

THERMAL BEHAVIOR OF THERMOTROPIC HYDROXYETHYL CELLULOSE ACETATE/POLYETHYLENE BLENDS

The thermal behavior of thermotropic hydroxythyl cellulose acetate (HECA)/polyethy-lene (PE) blends has been studied by DSC. It is found that the blends of HECA and PEare immiscible but the crystallization of PE is affected by HECA chains in the blends withmore than 50% HECA, which results in the subordinate crystallization of PE and the for-mation of imperfect structures in the PE crystals. The imperfection of PE crystals in theblends can be eliminated after annealing at 393K.     

Application of Single—labelled Probe—primer in PCR Amplification to the Detection of Hepatitis B Virus DNA

A new method based on the incorporation of a single-lablled probe-primer into polymerase chain reaction(PCR) for the detection of PCR-amplified DNA in a closed system is reported.The probeprimerc consists of a specific probe sequence on the 5‘‘‘‘‘‘‘‘-end and a primer sequence on the 3‘‘‘‘‘‘‘‘-end.A flurophore is located at the 5‘‘‘‘‘‘‘‘end.The primeR-quencher is an oligonucleotide,which is complementary to the probe sequence of probe-primer and labelled with a quencher at the 3‘‘‘‘‘‘‘‘-end.In the duplex formed by probe-primer and primer-quencher.the fluorophore and quencher are kept in close proximity to each other.Therefore the fluorescence is quenched.During PCR amplificatio,the specific probe sequence of probeprimer binds to its complement within the same strand of DNA,and is cleaved by Taq DNA polymerase,resulting in the restoration of fluorescence.This system has the same energy transfer mechanism as molecular beacons,and a good quenching effciency can be ensured.Following optimization of PCR conditions,this method was used to detect hepatitis b virus(HBV) dna in patient sera.This technology eliminates the risk of carry-over contamination,simplifies the amplification assay and opens up new possibilities for the real-time detection of the amplified DNA.     

The preparation of cationic folic acid and its application in drug delivery system

The cationic folic acid（CFA） was prepared by introducing triethylenetetramine into folic acid with EDCI/NHS and characterized by IR, NMR and mass spectra. It was found that approximately one of two carboxyls in the folic acid molecule was substituted to form CFA. The conversion of γ-carboxyl is found to be 59% higher than 30% of γ-carboxyl. The CFA and doxorubicin hydrochloride can be loaded on the ionic shell of PTX-encapsulated micelle to form CFA loaded binary drug carrier via static interaction in aqueous solutions. The successful loading was demonstrated by zeta potential measurement and the drug load amount（DLA） of CFA was measured by HPLC. In vitro cytotoxicity results revealed the CFA drug carrier showed higher cytotoxicity to cancer cell MDA-MB-321 than the binary drug carrier without CFA and the positive control, while it showed lower cytotoxicity to normal cell HUVEC than the positive control, and similar cytotoxicity with the binary drug carrier without CFA. These results as well as confocal laser scanning microscopy observation indicate the synthesized CFA drug carrier possesses active tumor-targeting property.     

Kinetics and Mechanism of the Photo-oxidation of Thiophene by O2 Adsorbed on Molecular Sieves

Photochemical oxidation of thiophene in n-octane/water extraction system using O2 as oxidant was studied. The reaction mechanism ofthiophene oxidation was proposed. Results obtained here can be used as the reference for the oxidative desulfurization of gasoline because thiophene is one of the main components containing sulfur in fluid catalytic cracking gasoline. Thiophene dissolved in n-octane was photodecomposed and removed into the water phase at ambient temperature and atmospheric pressure. A 500 W high-pressure mercury lamp （main wave length 365 nm, 0.22 kW/m） was used as light source for irradiation, and air was introduced by a gas pump to supply O2. Thiophene can be photo-oxidized to sulfone, oxalic acid, SO4^2-, and CO2. The desulfurization yield of thiophene in n-octane is 58.9% under photo-irradiation for 5 h under the conditions of air flow at 150 mL/min and V（water）：V（n-octane）=1：1. It can be improved to 92.3% by adding 0.15 g zeoliteartificial into 100 mL reaction system, which is the adsorbent for O2 and thiophene. And under such conditions, the photo-oxidation kinetics of thiophene with O2/zeoliteartificial is first-order with an apparent rate constant of 0.5047 h^-1 and a half-time of 1.37 h. The sulfur content can be depressed from 800 μL/L to less than 62 μL/L.