Biomedical subjects

R Zheng

Publications and source records attributed to R Zheng.

At least 55 records · Page 3Linked to original sources

PubMed1999

Chemokines in infection and inflammatory disease--the Royal Society of Medicine.

This well-organized meeting covered many clinical and academic areas related to chemokines and their receptors. Speakers from both sides of the Atlantic attended, including distinguished scientists and clinicians from both academic and pharmaceutical institutions. Although the meeting did not reveal sensational research data, it provided a forum in which attendees could express their views, using their research results as confirmation. Currently, the pathogenic and therapeutic roles of chemokines, and in particular their receptors, are clearer, although they are still not fully understood in HIV/AIDS and inflammatory diseases, such as rheumatoid arthritis and asthma. Progress will only be made with further research in order to reveal the importance and therapeutic value of these molecules. In this sense, the meeting provided good food for thought, with suggestions of possible targets and the discussion of various applications of research findings.

Journal Article↗

PubMed1998

Structural symmetry: the three-dimensional structure of Haemophilus influenzae diaminopimelate epimerase.

The Haemophilus influenzae diaminopimelate epimerase was cloned, expressed, purified, and crystallized in the C2221 space group (a = 102.1 A, b = 115.4 A, c = 66.3 A, alpha = beta = gamma = 90 degrees). The three-dimensional structure was solved to 2.7 A using a single Pt derivative and the Se-Met-substituted enzyme to a conventional R factor of 19.0% (Rfree = 24.2%). The 274 amino acid enzyme consists of two structurally homologous domains, each containing eight beta-strands and two alpha-helices. Diaminopimelate epimerase is a representative of the PLP-independent amino acid racemases, for which no structure has yet been determined and substantial evidence exists supporting the role of two cysteine residues as the catalytic acid and base. Cys73 of the amino terminal domain is found in disulfide linkage, at the domain interface, with Cys217 of the carboxy terminal domain, and we suggest that these two cysteine residues in the reduced, active enzyme function as the acid and base in the mechanism.

Amino Acid Isomerases↗

PubMed1998

Hydrolysis of N-succinyl-L,L-diaminopimelic acid by the Haemophilus influenzae dapE-encoded desuccinylase: metal activation, solvent isotope effects, and kinetic mechanism.

Hydrolysis of N-succinyl-L,L-diaminopimelic acid by the dapE-encoded desuccinylase is required for the bacterial synthesis of lysine and meso-diaminopimelic acid. We have investigated the catalytic mechanism of the recombinant enzyme from Haemophilus influenzae. The desuccinylase was overexpressed in Escherichia coli and purified to homogeneity. Steady-state kinetic experiments verified that the enzyme is metal-dependent, with a Km for N-succinyl-L,L-diaminopimelic acid of 1.3 mM and a turnover number of 200 s-1 in the presence of zinc. The maximal velocity was independent of pH above 7 but decreased with a slope of 1 below pH 7. The pH dependence of V/K was bell-shaped with apparent pKs of 6.5 and 8.3. Both L,L- and D,L-diaminopimelic acid were competitive inhibitors of the substrate, but d,d-diaminopimelic acid was not. Solvent kinetic isotope effect studies yielded inverse isotope effects, with values for D2OV/K of 0.62 and D2OV of 0.78. Determination of metal stoichiometry by ICP-AES indicated one tightly bound metal ion, while sequence homologies suggest the presence of two metal binding sites. On the basis of these observations, we propose a chemical mechanism for this metalloenzyme, which has a number of important structurally defined homologues.

Amidohydrolases↗

PubMed1998

Solution structure of the His12 --> Cys mutant of the N-terminal zinc binding domain of HIV-1 integrase complexed to cadmium.

The solution structure of His12 --> Cys mutant of the N-terminal zinc binding domain (residues 1-55; IN(1-55)) of HIV-1 integrase complexed to cadmium has been solved by multidimensional heteronuclear NMR spectroscopy. The overall structure is very similar to that of the wild-type N-terminal domain complexed to zinc. In contrast to the wild-type domain, however, which exists in two interconverting conformational states arising from different modes of coordination of the two histidine side chains to the metal, the cadmium complex of the His12 --> Cys mutant exists in only a single form at low pH. The conformation of the polypeptide chain encompassing residues 10-18 is intermediate between the two forms of the wild-type complex.

Binding Sites↗

PubMed1998

Role of superoxide anion on the proliferation and c-Ha-ras or p53 expression in prostate cancer cell line PC3.

The purpose of this study was to investigate the role of superoxide anion (02-*) in the regulation of p53 or c-Ha-ras expression and proliferation in the prostate cancer cell line PC3. Cell proliferation was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in the presence of O2-*, basic fibroblast growth factor (bFGF) or their combination. p53 or C-Ha-ras expression in the cells treated with O2-* was assayed by fluorescence in situ hybridization (FISH). The proliferation was significantly inhibited by O2-* in a concentration-dependent manner ranging from 9 to 36 micromol/l nicotinamide adenine dinucleotid (NADH) combined with 2-8 micromol/l N-methylphenazonium methyl sulfate (PMS). Enhancement of proliferation by 2 ng/ml bFGF was significantly inhibited by O2-*. Although O2-* was not able to alter c-Ha-ras gene expression, O2-* at the concentrations of 18 micromol/l NADH and 4 micromol/l PMS upregulated the expression of p53. O2-* may modulate proliferation and gene expression in PC3 cells.

Anions↗

PubMed1998

Solution structure of the cellular factor BAF responsible for protecting retroviral DNA from autointegration.

The solution structure of the human barrier-to-autointegration factor, BAF, a 21,000 Mr dimer, has been solved by NMR, including extensive use of dipolar couplings which provide a priori long range structural information. BAF is a highly evolutionarily conserved DNA binding protein that is responsible for inhibiting autointegration of retroviral DNA, thereby promoting integration of retroviral DNA into the host chromosome. BAF is largely helical, and each subunit is composed of five helices. The dimer is elongated in shape and the dimer interface comprises principally hydrophobic contacts supplemented by a single salt bridge. Despite the absence of any sequence similarity to any other known protein family, the topology of helices 3-5 is similar to that of a number of DNA binding proteins, with helices 4 and 5 constituting a helix-turn-helix motif. A model for the interaction of BAF with DNA that is consistent with structural and mutagenesis data is proposed.

Amino Acid Sequence↗

PubMed1998

Mechanisms of isoniazid resistance in Mycobacterium tuberculosis: enzymatic characterization of enoyl reductase mutants identified in isoniazid-resistant clinical isolates.

Mutants in the structural gene of the inhA-encoded NADH-dependent 2-trans enoyl-acyl carrier protein reductase were identified from isoniazid-resistant clinical isolates of Mycobacterium tuberculosis. Recombinant InhA proteins with defined single amino acid replacements were expressed in Escherichia coli and purified to homogeneity. Steady-state kinetic parameters for wild type (WT) and I16T, I21V, I47T, and I95P mutants of the enoyl reductase were measured spectrophotometrically. NADH binding to WT and I16T, I21V, I47T, S94A, and I95P mutant reductases were determined by fluorescence spectroscopy and demonstrated that all mutant enzymes had reduced NADH affinity and that NADH binding to all mutants was cooperative as compared with the hyperbolic binding of NADH to the WT enzyme. Since KatG-produced electrophilic derivatives of isoniazid have been suggested to inactivate the enoyl reductase-NADH complex, the kinetics of inactivation for the WT and I21V and I95P mutants was determined. Both mutations resulted in significantly increased values for the apparent first-order rate constant of inactivation.

Antitubercular Agents↗

PubMed1998

[An experimental research of recombinant human epidermal growth factor on corneal wound healing].

OBJECTIVE: To investigate the effects of recombinant human epidermal growth factor (rhEGF) eye drops on corneal wound healing. METHODS: Twenty-four white rabbits were randomly divided into 4 groups, 6 rabbits 12 eyes each. Anterior keratectomy of 8 mm in diameter and 1/3 cornea in thickness was performed on each eye. Each of the following concentrations of rhEGF: 1, 10, 100 microg/ml eye drops or normal saline (control) was applied four times daily for a week respectively for one group. The wound area was determined by computer imaging analysis. RESULTS: The mean epithelial healing rate of rhEGF 1, 10, 100 microg/ml groups was 9.31, 9.96, 9.31 mm(2)/day respectively, significantly greater than 8.11 mm(2)/day of the control group. The action of rhEGF of 10 microg/ml was somewhat better than that of 1 or 100 microg/ml, and no significant difference was noticed among the three rhEGF groups. Moderate inflammation and corneal neovascularization were induced in the rhEGF 100 microg/ml treated group. CONCLUSION: rhEGF 1 - 10 microg/ml can accelerate corneal wound healing in the rabbit with no adverse side-effects. It may be used to treat serious corneal trauma and ulcer clinically.

Animals↗

PubMed1998

[The study of changes of basement membrane in gingivae of adult periodontitis].

The major components of basement membrane including laminin and type IV collagen were studied by immunohistochemistry in 20 gingival specimens from patients with moderate to severe adult periodontitis and 10 normal gingival specimens combing with electron microscopic and PAS analysis. The results showed that laminin and type IV collagen were both present and stained with a continuous liner pattern in oral sulcular epithelium of the normal gingivae, while the pocket epithelial alterations of the periodontitis occurred, which included thinning, interruption, partial or complete absence of basement membrane involving one or more rete pegs and fragmentations. All these alterations may play an important role in the pathogenesis of adult periodontitis.

Adult↗

PubMed1998

[Determination of Ni and Pd in white karat gold jewellery by the EDXRF extrapolate-regression method].

The EDXRF extrapolate-regression method described in this paper combines regression method with the fundamental formula of fluorescence intensity. The contents of Ni and Pd in white karat gold jewellery were calculated theoretically according to the spectrum of the sample. The content of gold was deternined without standards. The precision was 0.1% and the deviation was 0.3% compared with AA.

English Abstract↗

PubMed1997

Three-dimensional structure of Escherichia coli dihydrodipicolinate reductase in complex with NADH and the inhibitor 2,6-pyridinedicarboxylate.

Dihydrodipicolinate reductase catalyzes the NAD(P)H-dependent reduction of the alpha,beta-unsaturated cyclic imine dihydrodipicolinate to form the cyclic imine tetrahydrodipicolinate. The enzyme is a component of the biosynthetic pathway that leads to diaminopimelate and lysine in bacteria and higher plants. Because these pathways are unique to microorganisms and plants, they may represent attractive targets for new antimicrobial or herbicidal compounds. The three-dimensional structure of the ternary complex of Escherichia coli dihydrodipicolinate reductase with NADH and the inhibitor 2,6-pyridinedicarboxylate has been solved using a combination of molecular replacement and noncrystallographic symmetry averaging procedures and refined against 2.6 A resolution data to a crystallographic R-factor of 21.4% (Rfree is 29.7%). The native enzyme is a 120 000 molecular weight tetramer of identical subunits. The refined crystallographic model contains a tetramer, three molecules of NADH, three molecules of inhibitor, one phosphate ion, and 186 water molecules per asymmetric unit. Each subunit consists of two domains connected by two flexible hinge regions. While three of the four subunits of the tetramer have a closed conformation, in which the nicotinamide ring of the cofactor bound to the N-terminal domain and the reducible carbon of the substrate bound to the substrate binding domain are about 3.5 A away, the fourth subunit is unliganded and shows an open conformation, suggesting that the enzyme undergoes a major conformational change upon binding of both substrates. The residues involved in binding of the inhibitor and the residues involved in catalysis have been identified on the basis of the three-dimensional structure. Site-directed mutants have been used to further characterize the role of these residues in binding and catalysis. A chemical mechanism for the enzyme, based on these and previously reported data, is proposed.

Binding Sites↗

PubMed1997

Repair effect of phenylpropanoid glycosides on thymine radical anion induced by pulse radiolysis.

Repair effects on thymine radical anion by six phenylpropanoid glycosides (PPGs), isolated from Pedicularis species, were studied using pulse radiolysis method. The thymine radical anion was produced by the reaction of hydrated electron with thymine. PPGs were added into the thymine solution saturated with N(2). Kinetic analysis showed that transient absorption spectrum of thymine radical anion formed at first, and then after several microseconds of pulse radiolysis changed to that of PPG radical anion. The evidence indicated that thymine radical anion was repaired through one-electron transfer between the radical anion and PPG. Electrophilic phenyl-substituted unsaturated carboxylic group containing in PPGs' structure was able to capture electron from thymine radical anion before it undergo reversible protonation. The reaction rate constants of electron transfer from thymine radical anion to PPGs were within 1.16-2.29 x 10(9) dm(3) mol(-1) s(-1).

Journal Article↗

PubMed1997

The effects of CD3, CD4 and CD28 signaling on lymphocytes during human immunodeficiency virus-1 infection.

We investigated the effects of early human immunodeficiency virus-1 infection (HIV-1) on CD4- and CD28-mediated co-signaling of the T cell receptor (TCR)/CD3 complex in peripheral blood lymphocytes (PBL). CD4 ligation either alone or in conjunction with TCR occupancy resulted in abrogated signaling shown by impaired co-association of the tyrosine kinase ZAP-70 with the CD3-zeta chains in virally infected PBL. In addition, down-regulation of CD4-associated TCR signaling resulted in diminished tyrosine phosphorylation of mitogen-activated protein kinase (MAPK), a serine threonine kinase which is critically involved in the regulation of transcription factors. Furthermore, these aberrant CD4-driven signals rendered HIV-1-infected PBL susceptible to activation-induced cell death. By contrast, cross-linking of the TCR/CD3 complex with the CD28 receptor improved tyrosine phosphorylation of MAPK and salvaged infected PBL from activation-induced cell death. Our data demonstrate the importance of appropriate CD3, CD4 and CD28 co-stimulatory function to prevent apoptosis. The CD4-mediated signaling defects of the TCR could contribute to the loss of immunocompetent cells during HIV-1 infection via activation-induced cell death, whereas stimulation through the CD28 pathway could reverse these detrimental effects.

Apoptosis↗

PubMed1997

In vitro effects of prostaglandin E2 or indomethacin on the proliferation of lymphokine-activated killer cells and their cytotoxicity against bladder tumor cells in patients with bladder cancer.

PURPOSE: To investigate the combined effects of interleukin-2 (IL-2) with either prostaglandin E2 (PGE2) or indomethacin (IM) on the proliferation and cytolysis of bladder tumor cells by lymphokine-activated killer (LAK) cells in patients with bladder cancer. METHODS: LAK cell proliferation was assayed in the presence of various concentrations of either PGE2 or IM by cell counting. Bladder cancer cell lines BIU-87, EJ and bladder tumor cells (BTC) from the patients were cultured as target cells, and the cytotoxicity of LAK cells was determined by 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. In addition, PGE2 in samples of conditioned medium from bladder cancer cells or peripheral blood mononuclear cells (PBMC) as well as plasma from 21 patients with bladder cancer and 20 healthy donors were determined by radioimmunoassay (RIA). RESULTS: The proliferation of LAK cells induced by IL-2 was inhibited by PGE2 (0.05 to 5 ng/mL) in concentration-dependent manner. The enhanced growth of LAK cells was observed at certain concentrations of IM (100-400 ng/mL) from 48 to 96 h. Pretreatment of LAK cells with IM (200 ng/mL) significantly enhanced cytotoxicity against BIU-87, EJ cells, or BTC. More PGE2 was present in conditioned medium from BIU-87 cells than in the conditioned medium from PBMC. CONCLUSIONS: These studies indicate that LAK cell proliferation induced by IL-2 in patients with bladder cancer is inhibited by PGE2 produced by PBMC and bladder cancer cells. This inhibition can be overcome by IM, which may be of use in immunotherapy of bladder cancer.

Carcinoma, Transitional Cell↗

PubMed1997

Solution structure of the N-terminal zinc binding domain of HIV-1 integrase.

The solution structure of the N-terminal zinc binding domain (residues 1-55; IN1-55) of HIV-1 integrase has been solved by NMR spectroscopy. IN1-55 is dimeric, and each monomer comprises four helices with the zinc tetrahedrally coordinated to His 12, His 16, Cys 40 and Cys 43. IN1-55 exists in two interconverting conformational states that differ with regard to the coordination of the two histidine side chains to zinc. The different histidine arrangements are associated with large conformational differences in the polypeptide backbone (residues 9-18) around the coordinating histidines. The dimer interface is predominantly hydrophobic and is formed by the packing of the N-terminal end of helix 1, and helices 3 and 4. The monomer fold is remarkably similar to that of a number of helical DNA binding proteins containing a helix-turn-helix (HTH) motif with helices 2 and 3 of IN1-55 corresponding to the HTH motif. In contrast to the DNA binding proteins where the second helix of the HTH motif is employed for DNA recognition, IN1-55 uses this helix for dimerization.

Bacterial Proteins↗

PubMed1997

[The free radicals and nickel carcinogenesis].

The relationship between free radicals and the carcinogenesis of nickel and its compounds was summarized. Ni(II) could enhance the production of free radicals mainly in three pathways: Fenton reaction, enhancing lipid peroxidation and decreasing cellular antioxidantive defense system. Thus free radacals lead cell carcinogenesis. It suggested that the carcinogenesis of Ni(II) was mainly related to the effect of free radicals.

Animals↗

PubMed1997

Differentiation of human gastric adenocarcinoma cell line MGc80-3 induced by verbascoside.

Verbascoside is a natural antioxidant extracted from Pedicularis striata Pall (Jueyehesen). After being treated with 20 mumol/l verbascoside, the growth curve and mitotic index of human gastric adenocarcinoma MGc80-3 cells decreased remarkably, cell doubling time was delayed, the cellular growth inhibitory rate amounted to 53.2%, cell surface charge assayed by cell electrophoresis obviously changed, the electrophoresis rate dropped from 3.51 microns/s/v/cm to 2.74, i.e., the percent of retardation reached 28.4%. There was a 75% decrease of the tumorigenicity for the treated cells compared with the untreated cells inoculated subcutaneously in BALB/C nude mice. Scanning electron microscopy revealed that the microvilli on the surface of treated cells had been reduced obviously. It confirmed that verbascoside, similar to DMSO, could reverse MGc80-3 cells' malignant phenotypic characteristics and induced redifferentiation of MGc80-3 cells.

Adenocarcinoma↗

PubMed1997

Effect of TNF-alpha and IFN-alpha on the proliferation and cytotoxicity of lymphokine-activated killer cells in patients with bladder cancer.

OBJECTIVE: To investigate the effects of interleukin-2 (IL-2) combined with either tumor necrosis factor-alpha (TNF-alpha) or alpha-interferon (IFN-alpha) on the proliferation and cytolysis to bladder tumor cells of lymphokine-activated killer (LAK) cells in patients with bladder cancer. METHODS: LAK cells were generated by ficoll-paque density-centrifugation from 21 patients with bladder cancer and cultured in medium containing IL-2. LAK cell proliferation was assayed in the presence of various concentrations of either TNF-alpha or IFN-alpha by cell count in 96-well plates. Bladder cancer cell lines BIU-87 and EJ were cultured as target cells and the cytotoxicity of LAK cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide (MTT) assay. RESULTS: The proliferation of LAK cells induced by IL-2 was enhanced by TNF-alpha in a dose-responsive fashion. The direct growth support for the LAK cells was also observed with IFN-alpha at the concentration of 1000 U/ml after 48 hours of culture. TNF-alpha (5000 U/ml) resulted in an increase in the cytotoxicity of LAK cells to BIU-87 and EJ cells. However, the change of cytotoxicity of LAK cells treated with IFN-alpha was not statistically significant. CONCLUSIONS: TNF-alpha and IFN-alpha enhance the proliferation and activation of LAK cells and influence their antitumor cytotoxicity in patients with bladder cancer.

Carcinoma, Transitional Cell↗