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Biomedical subjects

L Qin

Publications and source records attributed to L Qin.

At least 127 records · Page 7Linked to original sources
PubMed1998

[Effects of modulation of abscisic acid during somatic embryogenesis in Lycium barbarum L].

Using Enzyme Linked Immunosorbent Assay (ELISA) method, we determined the ABA contents of different stages in somatic embryogenesis. The results showed that endogenous ABA contents increased to maximum value twice during somatic embryogenesis. After first maximum value of ABA contents embryogenic cells were observed in callus, and simultaneously, there was a specific protein of somatic embryogenesis investigated by SDS-PAGE. This protein accumulates preferentially in embryogenic callus but not in transferred callus. So it is suggested that ABA could promote the expression of specific genes and the synthesis of embryogenic protein during somatic embryogenesis in Lycium barbarum L. and ABA play an important role in globular stage as well. In addition, treatment of non-embryogenic activity callus with 4 mumol/L exogenous ABA could stimulate somatic embryogenesis. And the ABA function mechanism in relation to somatic embryogenesis was discussed.

Abscisic Acid↗
PubMed1998

[Study on the design and application of the contact Nd:YAG laser scalper].

A method of designing the contact laser scalper is presented. The calculations using this method is in good agreement with the measured results in laboratory. Some operations on rabbits using the newly developed contact Nd:YAG laser scalper are performed and shown that the contact Nd:YAG laser is a promising medical equipment.

Aluminum Oxide↗
PubMed1997

Promoter attenuation in gene therapy: interferon-gamma and tumor necrosis factor-alpha inhibit transgene expression.

One of the major limitations to current gene therapy is the low-level and transient vector gene expression due to poorly defined mechanisms, possibly including promoter attenuation or extinction. Because the application of gene therapy vectors in vivo induces cytokine production through specific or nonspecific immune responses, we hypothesized that cytokine-mediated signals may alter vector gene expression. Our data indicate that the cytokines interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) inhibit transgene expression from certain widely used viral promoters/enhancers (cytomegalovirus, Rous sarcoma virus, simian virus 40, Moloney murine leukemia virus long terminal repeat) delivered by adenoviral, retroviral or plasmid vectors in vitro. A constitutive cellular promoter (beta-actin) is less sensitive to these cytokine effects. Inhibition is at the mRNA level and cytokines do not cause vector DNA degradation, inhibit total cellular protein synthesis, or kill infected/transfected cells. Administration of neutralizing anti-IFN-gamma monoclonal antibody results in enhanced transgene expression in vivo. Thus, standard gene therapy vectors in current use may be improved by altering cytokine-responsive regulatory elements. Determination of the mechanisms involved in cytokine-regulated vector gene expression may improve the understanding of the cellular disposition of vectors for gene transfer and gene therapy.

Adenoviruses, Human↗
PubMed1997

Purification and cloning of hepatocyte growth factor activator inhibitor type 2, a Kunitz-type serine protease inhibitor.

Hepatocyte growth factor (HGF) activator is a serine protease responsible for proteolytic activation of HGF in response to tissue injury and thus plays an important role in the regulation of biological functions of HGF in regenerating tissue. We previously purified an inhibitor of HGF activator (HGF activator inhibitor type 1, HAI-1) from the conditioned medium of a human stomach carcinoma cell line MKN45 and cloned its cDNA. HAI-1 is a novel member of the Kunitz family of serine protease inhibitors. In the present study, we purified a second type of HGF activator inhibitor (HAI-2) from the conditioned medium of MKN45 cells and molecularly cloned its cDNA. The cDNA sequence revealed that HAI-2 is derived from a precursor protein of 252 amino acids and contains two Kunitz domains, indicating that HAI-2 is also a member of the Kunitz family of serine protease inhibitors. The primary translation product of HAI-2 has a hydrophobic sequence in the COOH-terminal region, suggesting that, like HAI-1, HAI-2 is produced in a membrane-associated form and secreted in a proteolytically truncated form. Because HAI-2 and HAI-1 are potent inhibitors specific for HGF activator, they may be involved in regulation of proteolytic activation of HGF in injured tissues.

Amino Acid Sequence↗
PubMed1997

Adenovirus-mediated gene transfer of viral interleukin-10 inhibits the immune response to both alloantigen and adenoviral antigen.

Although adenoviral vectors are attractive for gene transfer, their effectiveness is limited by host antiviral immune responses. In this study, we determined if host antiallograft and antiviral immunity could be diminished with an adenoviral vector encoding the immunosuppressive cytokine viral interleukin-10 (vIL-10). AdSV40vIL-10, a vIL-10-expressing adenoviral vector with an SV40 promoter, induced significant prolongation of murine cardiac allograft survival to 32.2 +/- 1.7 days compared to 14.2 +/- 1.0 days for controls (p < 0.01). This effect was specific for vIL-10 encoding vector and could be inhibited by anti-vIL-10 monoclonal antibody (mAb). In vivo administration of adenovirus facilitated the generation of adenovirus-specific cytotoxic T lymphocytes (CTL), whereas treatment with AdSV40vIL-10 prevented CTL priming and generation of virus-specific immunity. AdSV40vIL-10 also induced extended expression of a beta-galactosidase reporter from a co-injected LacZ-encoding adenoviral vector. These results demonstrate that adenovirus-mediated gene transfer and expression of vIL-10 prolong allograft survival and inhibit the immune response to adenoviral antigens, thereby improving the persistence of the vector and extending transgene expression. The efficacy of adenoviral vectors can be improved by incorporating immunosuppressive genes into the vector.

Adenoviridae↗
PubMed1997

Hepatocyte growth factor activator inhibitor, a novel Kunitz-type serine protease inhibitor.

Hepatocyte growth factor (HGF) activator is a serine protease that is produced and secreted by the liver and circulates in the blood as an inactive zymogen. In response to tissue injury, the HGF activator zymogen is converted to the active form by limited proteolysis. The activated HGF activator converts an inactive single chain precursor of HGF to a biologically active heterodimer in injured tissue. The activated HGF may be involved in the regeneration of the injured tissue. In this study, we purified an inhibitor of HGF activator from the conditioned medium of a human MKN45 stomach carcinoma cell line and molecularly cloned its cDNA. The sequence of the cDNA revealed that the inhibitor has two well defined Kunitz domains, suggesting that the inhibitor is a member of the Kunitz family of serine protease inhibitors. The sequence also showed that the primary translation product of the inhibitor has a hydrophobic sequence at the COOH-terminal region. Inhibitory activity toward HGF activator was detected in the membrane fraction as well as in the conditioned medium of MKN45 cells. These results suggest that the inhibitor may be produced as a membrane-associated form and secreted by the producing cells as a proteolytically truncated form.

Amino Acid Sequence↗
PubMed1997

Electrical stimulation prevents immobilization atrophy in skeletal muscle of rabbits.

OBJECTIVE: To investigate the effect of unilateral cast immobilization with and without surface electrical stimulation (ES) on the tibialis anterior (TA) muscle of rabbits. DESIGN: Prospective randomized trial. SETTING: University medical school. ANIMALS: 53 New Zealand White rabbits (aged 54 to 63 days, weight 1.73 to 1.91 kg). METHODS AND INTERVENTION: Random assignment, for a 3-week period, to one of four groups: C group (control group), I group (immobilization group), S group (group of electrical stimulation which was stimulated isometrically at 50 Hz, 30 minutes per day, 5 times a week), and IS group (immobilization group which, like the S-group, received electrical stimulation). OUTCOME MEASURES: Muscle wet wight, muscle fiber cross-sectional area, muscle fiber types, and muscle capillary supply. RESULTS: Muscle wet weight decreased significantly in the I group by 19% (p < or = .05), with a corresponding significant reduction in the total muscle fiber cross-sectional area of 26% (p < or = .05). No significant changes were observed in muscle wet weight and muscle fiber cross-sectional area in the S and IS groups. Interstitial fibrosis was observed in the I group and occasionally in the IS group. No significant changes in the total number of muscle fiber types I and II were found in all experimental groups. The capillary supply of the S and IS groups did not change significantly. However, capillary-to-fiber ratio was significantly reduced by 20% with a simultaneously nonsignificant increase in capillary density (capillaries/mm2) of 11% (p > .05) in the I group. Furthermore, muscle fiber regeneration was observed predominantly in the I group. CONCLUSIONS: In this experimental model, ES effectively prevented immobilization-induced muscle atrophy by minimizing reduction of muscle fiber cross-sectional area, interstitial fibrosis, and impaired blood supply.

Animals↗
PubMed1997

Transcriptional repression by Oshox1, a novel homeodomain leucine zipper protein from rice.

This paper describes the characterization of Oshox1, a cDNA clone from rice encoding a member of the homeodomain-leucine zipper (HD-Zip) class of putative transcription factors. Oshox1 maps to chromosome 10 and belongs to a family of related rice genes. Two-hybrid assays showed that Oshox1 protein can homodimerize, but can also form heterodimers with an Arabidopsis HD-Zip protein. This suggests that protein-protein interactions may also occur between different HD-Zip proteins in rice, which would provide enormous versatility for generating specific gene-control mechanisms. Oshox1 mRNA could be detected in various rice tissues at different developmental stages, with highest levels in embryos, shoots of seedlings, and leaves of mature plants. Transgenic expression of Oshox1 in Arabidopsis retarded growth and affected leaf size and shape, indicative of a role as developmental regulator. In vitro and in vivo DNA-binding studies revealed that Oshox1 interacts with the pseudopalindromic sequence CAAT(C/G)ATTG, confirming that the protein represents a transcription factor. Oshox1 was found to repress reporter gene activity in rice suspension cells, most likely by a mechanism of active transcriptional repression. Repression was strictly dependent on the presence of upstream Oshox1 binding sites in the reporter gene constructs and a function of the N-terminal region of Oshox1, preceding the homeodomain.

Amino Acid Sequence↗
PubMed1997

Retinoid X receptor-specific ligands synergistically upregulate 1, 25-dihydroxyvitamin D3-dependent transcription in epidermal keratinocytes in vitro and in vivo.

We have examined the mechanism by which endogenous retinoid X receptor (RXR), vitamin D3 receptor (VDR), and cognate ligands regulate nuclear 1,25-dihydroxyvitamin D3 (D3) signaling in epidermal keratinocytes from skin, a physiologic D3 target. In vitro, RXR and VDR-specific antibodies identified endogenous RXR and VDR bound to a vitamin D3-responsive element (DR3) as heterodimers (VDR-RXR). In cultured keratinocytes, 9-cis retinoic acid (9cRA), a panagonist for RXR and retinoic acid receptor (RAR), and an RXR-selective agonist, SR11237, synergized with D3 to activate DR3 via endogenous as well as overexpressed VDR-RXR, whereas both of these RXR agonists alone were ineffective. In contrast, SR11237 did not synergize with but antagonized an RAR-selective ligand activation of a retinoic acid-responsive element (DR5) via endogenous RAR-RXR. Furthermore, expression of RXR mutated in transactivation domain AF-2 inhibited endogenous VDR-RXR activity over DR3. This mutant efficiently bound to DR3 as VDR-RXR but showed reduced capacity to transactivate DR3 in response to D3 and SR11237. In vivo, D3 and SR11237 synergistically induced the naturally occurring D3-responsive 24-hydroxylase gene in epidermis of mouse skin, whereas SR11237 alone was ineffective. Our data suggest that allosteric changes caused by VDR in DR3-bound VDR-RXR do not block access of ligands to RXR. RXR ligand-induced conformational changes permit VDR-RXR, via both VDR and RXR activation function domains, to mediate maximal D3 signaling in keratinocytes.

Animals↗
PubMed1996

The roles of CD11/CD18 and ICAM-1 in acute Pseudomonas aeruginosa-induced pneumonia in mice.

Neutrophil accumulation in response to Pseudomonas aeruginosa in the lungs is mediated through CD11/CD18. This study determined the roles of CD11a, CD11b, and intercellular adhesion molecule (ICAM)-1 in P. aeruginosa-induced pneumonia and compared the function of ICAM-1 using Abs or ICAM-1 mutant mice. Anesthetized BALB/c mice pretreated with either Abs against CD11a, CD11b, ICAM-1, or rat IgG received intratracheal instillation of P. aeruginosa for 4 h. In other studies, ICAM-1 mutant and wild-type mice received either anti-ICAM-1 Ab or rat IgG followed by instillation of P. aeruginosa. The data show that Abs against CD11a, CD11b, and ICAM-1 in BALB/c mice inhibited neutrophil emigration by 79, 81, and 56%, respectively. ICAM-1 mutant mice showed no inhibition of neutrophil emigration compared with wild-type mice. Pretreatment with anti-ICAM-1 Ab inhibited neutrophil emigration in wild-type (129/SvxC57) mice by 67% but had no effect in ICAM-1 mutant mice, suggesting that the Ab was acting specifically through recognition of its Ag. We conclude that CD11a and CD11b are required for neutrophil emigration. The observed function of ICAM-1 varies depending on the method by which it is inhibited. Abs may overestimate function by altering other cellular functions or mutant mice may develop alternative pathways of emigration.

Acute Disease↗
PubMed1996

Gene transfer of transforming growth factor-beta 1 prolongs murine cardiac allograft survival by inhibiting cell-mediated immunity.

Delivery of immunosuppressants directly to allografts using gene transfer and gene therapy approaches may inhibit immune activation while avoiding the systemic toxicity of conventional immunosuppression. Cardiac grafts from allogeneic (C57BL/6, H-2b) donors were transplanted into CBA/J (H-2k) recipients in a heterotopic, non-vascularized model pSVTGF-beta 1, a plasmid encoding murine transforming growth factor-beta 1 (TGF-beta 1) under the control of an SV40 promoter, was directly injected into grafts at surgery and prolonged survival from 12.0 +/- 0.7 to 25.1 +/- 2.1 days (p < 0.001) in a dose-dependent manner. Plasmid gene transfer-induced immunosuppression was localized to the area of the graft because plasmid injected remote from the graft did not prolong allograft survival and systemic immunity was not influenced by local gene transfer. Limiting dilution analysis of graft-infiltrating cells demonstrated that gene transfer reduced the precursor frequency of donor-specific cytotoxic T lymphocytes (CTL) and activated and total interleukin-2 (IL-2) producing helper T lymphocytes (HTL) in graft-infiltrating cells, whereas CTL generation and HTL precursor frequency in splenic lymphocytes were not altered. Additional data revealed that gene transfer inhibited the priming of TH0 cells and the conversion of primed TH1 cells to activated cells without the participation of TH2 suppressors. These data demonstrate that gene transfer of plasmid DNA encoding TGF-beta 1 in vivo suppresses local T cell immunity, which prolongs allograft survival.

Animals↗
PubMed1996

Blockade of multiple costimulatory receptors induces hyporesponsiveness: inhibition of CD2 plus CD28 pathways.

T-lymphocyte activation requires engagement of the T cell receptor with antigen-major histocompatibility complex, and simultaneous ligation of costimulatory pathways via the lymphocyte receptors CD2 and CD28/ CTLA4. Anti-CD2 monoclonal antibody (mAb) blocks the interaction of the antigen-presenting cell receptor CD48 with its ligand CD2, whereas CTLA4Ig binds with high affinity to the antigen-presenting cell ligands B7-1 and B7-2, blocking their interaction with CD28/CTLA4. We tested the immunosuppressive effects of simultaneously blocking both costimulatory pathways. Using donor C57BL/6J (H2b) hearts transplanted to CBA/J (H2k) recipients, anti-CD2 mAb plus CTLA4Ig administered at the time of transplantation prolonged cardiac allograft mean survival time to >120 days compared with untreated controls (12.2+/-0.5 days, P<0.01), anti-CD2 mAb alone (24.8+/-1.0 days, P<0.01), or CTLA4Ig alone (55.0+/-2.0 days, P<0.01). Retransplantation of these recipients with donor-specific and third-party grafts demonstrated that hyporesponsiveness and tolerance were achieved. In vitro stimulation of lymphocytes from tolerant recipients with donor-specific alloantigen resulted in normal cytotoxic T lymphocyte and mixed lymphocyte reaction responses, showing that clonal deletion or anergy did not occur, but that graft adaptation or suppression likely helped to maintain long-term graft survival. In vitro combinations of anti-CD2 mAb and CTLA4Ig suppressed the generation of allogeneic cytotoxic T lymphocytes (58%) and the mixed lymphocyte reaction (36%); CTLA4Ig was more effective in this regard and the two agents were not synergistic. Anti-CD2 mAb and CTLA4Ig suppressed mitogen-driven proliferation in differential fashions, suggesting that they affected independent signaling pathways. Anti-CD2 mAb and CTLA4Ig also inhibited interleukin (IL)-2, IL-4, and IL-2 receptor (CD25). These data indicate that anti-CD2 mAb plus CTLA4Ig induces hyporesponsiveness and tolerance. The mechanism is likely related to the initial disruption of independent pathways of T-lymphocyte activation leading to antigen-specific long-term graft survival.

Abatacept↗
PubMed1996

A novel murine model for the assessment of human CD2-related reagents in vivo.

CD2 is a T cell surface glycoprotein that mediates both cell-cell adhesion and transmembrane signal transduction. To construct a model for the in vivo evaluation of human (h)CD2 function and hCD2-related reagents, hCD2 transgenic mice and murine (m)CD2 knockout mice were crossed, and the F2 generation selected for mCD2-hCD2+ animals by fluorescent flow cytometry. The mCD2-hCD2+ mice are healthy and have a normal distribution of mCD3, mCD4, and mCD8 in thymus, spleen, and lymph node. Therefore expression of the hCD2 transgene does not appear to disrupt normal T cell development. The functionality of hCD2 was demonstrated by T lymphocyte proliferation upon stimulation by combined anti-CD2 plus anti-CD2R (anti-T11(2) plus anti-T11(3)) mAbs. Anti-T11(2) plus anti-T11(3) anti-human CD2 mAbs also induced proliferation of mCD2+hCD2+ F1 lymphocytes, but not mCD2+hCD2- wild-type murine lymphocytes. Either an anti-murine or the human CD2 specific (anti-T11(1)) mAbs inhibited proliferation in alloantigen, PHA, or anti-CD3 mAb stimulated cultures and inhibited only cells bearing the appropriate cognate CD2. In vivo studies of immune function yielded results consistent with these in vitro assays. Thus, anti-T11(1) mAb suppressed contact sensitivity in vivo in the transgenic/knockout mice. mCD2-hCD2+ mice treated with anti-T11(1) or LFA-3 fusion proteins also showed significant prolongation of cardiac allograft survival. This prolongation was associated both with depletion and down-modulation of CD2 on remaining T cells. These data suggest that the transgenic/knockout mice provide a useful in vivo model for the assessment of hCD2-related reagents and CD2 function, free from any potential interactions with mCD2 and mCD2 ligands.

Animals↗
PubMed1996

Enforced interaction of one molecule of plastocyanin with two molecules of cytochrome c and an electron-transfer reaction involving the hydrophobic patch on the plastocyanin surface.

Laser flash photolysis is used to study the photoinduced electron-transfer reaction cyt(III)//pc(II) + 3Zncyt --> cyt(III)//pc(I) + Zincyt+ at pH 7.0 and 25 degrees. In the covalent (symbol//) complex cyt(III)//pc(II) the acidic patch in cupriplastocyanin is directly cross-linked to the basic patch in ferricytochrome c. The triplet state of zinc cytochrome c reduces the pc(II) moiety, not the cyt(III) moiety, of the covalent complex. The reaction is strictly bimolecular in the entire range of ionic strength studied, from 1.25 mM to 1.00 M. The two reactants interact only transiently, in a collisional complex, and do not form a persistent complex cyt(III)//pc(II)/Zncyt. Because noncovalent (symbol/) association of three separate protein molecules is far less probable than association of the covalent complex and another protein molecule, we conclude that, without the aid of covalent cross-links, one molecule of plastocyanin will not form a ternary complex with two molecules of cytochrome c, cyt/pc/cyt. Dependence of the rate constant on ionic strength is analyzed in terms of van Leeuwen theory of electrostatic interactions, which recognizes the importance of dipole moments of the proteins. This analysis shows that 3Zncyt reacts with the hydrophobic patch in the pc(II) moiety of the covalent complex cyt(III)//pc(II). At high ionic strength, at which electrostatic interactions are practically abolished, the blue copper site is reduced with approximately equal rates via the hydrophobic patch in the pc(II) moiety of the complex and via the acidic patch in free pc(II). This is evidence that the two distinct patches on the plastocyanin surface are comparable in their intrinsic "conductivity" for electrons coming to the copper site. Positively charged and electroneutral redox partners tend to react at the acidic patch (although not necessarily at the initial docking site in this broad patch) for electrostatic, not electronic, reasons. Earlier theorectical studies disagreed about the relative electronic conductivities of the two patches. This experimental study corroborates very recent theoretical studies that found the two patches to be comparable in the efficiency of electron transfer.

Animals↗