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Biomedical subjects

L Dong

Publications and source records attributed to L Dong.

At least 55 records · Page 3Linked to original sources
PubMed2001

[Determination of warfarin in plasma by HPLC and an investigation of monitoring patients after cardiac valve replacement].

OBJECTIVE: A simple HPLC method was established for the determination of warfarin in plasma to investigate the relationship between warfarin concentration and anticoagulant effect. METHODS: The mixture of dichloromethane and hexane (1:9) was used as extracting solvent for the plasma samples. The chromatographic separation was on C18 column with a mobile phase consisting of methanol and 50 mmol/L ammonium acetate buffer (pH2.5, 70:30). RESULTS: The calibration curve was linear within 50-2000 ng/ml. The extraction recoveries of warfarin were 78.0%-81.6%. The recoveries of methodology were 103.1%-106.5%. Inter-day and intra-day RSD were 2.33%-5.46% and 5.29%-7.73%, respectively. This method was used for determining warfarin in 70 patients after cardiac valve replacement. The results showed that 44 cases had their INR within the safety range (1.31-2.35) recommended to Chinese, and of them 37(84.1%) cases had a warfarin level at 616.2 +/- 154.8 ng/ml. CONCLUSION: This method is useful in monitoring warfarin concentration during anticoagulant therapy.

Anticoagulants↗
PubMed2001

Externalization and internalization of cardiac endothelin receptors during different phases of sepsis in rat.

OBJECTIVE: To study the redistribution of endothelin-1 (ET-1) receptors in two subcellular organelles, the sarcolemmal membrane and the light vesicle, of rat heart during the progression of sepsis. METHODS: Sepsis was induced by cecal ligation and puncture (CLP). ET1 receptor was assayed by using [125I]-ET1 binding. Marker enzyme activities, protein yield, and dry-to-wet weight ratio of cardiac membranes were measured. RESULTS: Septic rat heart exhibited two distinct phases: an initial hyperdynamic phase( 9h after CLP; early stage of sepsis) followed by a hypodynamic (18h after CLP, late stage of sepsis) phase. [125I]-ET1 binding study showed that during early stage of sepsis, the B(max) of ET1 receptors was increased by 30% in sarcolemma but decreased by 19% in light vesicles, while during late stage of sepsis, the B(max) was decreased by 24% in sarcolemma but increased by 38% in light vesicles. The total binding of sarcolemma and light vesicles was increased by 25% during early stage of sepsis but decreased by 17% during late stage of sepsis. CONCLUSIONS: These data indicated that ET1 receptors in the rat heart were externalized from light vesicles to sarcolemmal membranes during early hyperdynamic phase while internalized from surface membranes to intracellular compartment during late hypodynamic phase of sepsis.

Animals↗
PubMed2000

Founder's Award, Society for Biomaterials. Sixth World Biomaterials Congress 2000, Kamuela, HI,May 15-20, 2000. Really smart bioconjugates of smart polymers and receptor proteins.

Over the past 18 years we have been deeply involved with the synthesis and applications of stimuli-responsive polymer systems, especially polymer-biomolecule conjugates. This article summarizes our work with one of these conjugate systems, specifically polymer-protein conjugates. We include conjugates prepared by random polymer conjugation to lysine amino groups, and also those prepared by site-specific conjugation of the polymer to specific amino acid sites that are genetically engineered into the known amino acid sequence of the protein. We describe the preparation and properties of thermally sensitive random conjugates to enzymes and several affinity recognition proteins. We have also prepared site-specific conjugates to streptavidin with temperature-sensitive polymers, pH-sensitive polymers, and light-sensitive polymers. The preparation of these conjugates and their many fascinating applications are reviewed in this article.

Acrylamides↗
PubMed2000

The senescence-accelerated mouse shows aging-related defects in cellular but not humoral immunity against influenza virus infection.

The senescence-accelerated mouse (SAM) strain P1, which has a short life span, was adopted as a murine model for an investigation of the pathogenesis of viral infection in elderly adults. After intranasal inoculation with influenza A virus, the SAM-P1 mice showed a higher rate of mortality, with prolonged virus growth in the lungs. The increased susceptibility was associated with impaired activity of both NK cells and virus-specific cytotoxic T lymphocytes. The production of interferon-gamma and interleukin-12 was significantly restrained, which suggests a partial deficiency of the T helper (Th) 1 cells. In contrast, the immunologic activity of the Th2 cells appeared to be functionally normal, judging from the release of large amounts of interleukin-4 followed by production of appropriate amounts of influenza virus-specific antibody. It is suggested that the elicitation of cellular immunity is an important and effective procedure for protecting the elderly from influenza virus infection.

Aging↗
PubMed2000

Egg-adapted replication-restricted virus protects mice against lethal influenza.

Infection of mice with the egg-adapted (EA) strain of influenza virus was studied as a murine model of human live attenuated virus vaccine. The growth and spread of the EA virus in the mouse lungs were restricted, and only small inflammatory changes were detected in the respiratory tracts. Deletion and substitutions of amino acids were found in the hemagglutinin molecule of the EA virus, which were attributable to the reduced envelope fusion activity in virus multiplication cycles. Intranasal inoculation of mice with the EA virus induced specific IgG and IgA antibody production together with a specific cytotoxic T lymphocyte response. Immunized mice showed a distinct resistance to subsequent lethal challenge with the virulent influenza virus. These results indicate that the mutant virus loaded with a growth restriction in the respiratory tract is an appropriate candidate for a live attenuated vaccine.

Animals↗
PubMed2000

Application of a modified simplex method to the multivariable optimization of a new FIA system for the determination of osmium.

A methodology based on the coupling of experimental design and a modified simplex method is proposed for the optimization of a new flow injection-kinetic system for the spectrophotometric determination of Os (IV) with m-acetylchlorophosphonazo, which has for the first time been used as chromogenic reagent in the quantitative analysis of this element. An orthogonal array design is utilized to design the experimental protocol, in which six variables are varied simultaneously, and obtain the initial simplex using 25 experiments. A modified simplex method is applied to continuously optimize the data of the orthogonal array design; the search for optimum conditions of 6 variables using the modified simplex method required only 25 experiments. The efficiency and simplicity of the coupling of the experimental design and the modified simplex method are attractive for the development of new analytical methods. The method has been applied to the determination of Os (IV) in a refined ore as well as in a secondary alloy and provided satisfactory results.

Journal Article↗
PubMed2000

Preconditioning with cortical spreading depression decreases intraischemic cerebral glutamate levels and down-regulates excitatory amino acid transporters EAAT1 and EAAT2 from rat cerebal cortex plasma membranes.

We previously reported a 50% reduction in cortical infarct volume following transient focal cerebral ischemia in rats preconditioned 3 days earlier with cortical spreading depression (CSD). The mechanism of the protective effect of prior CSD remains unknown. Recent studies demonstrate reversal of excitatory amino acid transporters (EAATs) to be a principal cause for elevated extracellular glutamate levels during cerebral ischemia. The present study measured the effect of CSD preconditioning on (a) intraischemic glutamate levels and (b) regulation of glutamate transporters within the ischemic cortex of the rat. Three days following either CSD or sham preconditioning, rats were subjected to 200 min of focal cerebral ischemia, and extracellular glutamate concentration was measured by in vivo microdialysis. Cortical glutamate exposure decreased 70% from 1,772.4 +/- 1,469.2 microM-min in sham-treated (n = 8) to 569.0 +/- 707.8 microM-min in CSD-treated (n = 13) rats (p <0.05). The effect of CSD preconditioning on glutamate transporter levels in plasma membranes (PMs) prepared from rat cerebral cortex was assessed by western blot analysis. Down-regulation of the glial glutamate transporter isoforms EAAT2 and EAAT1 from the PM fraction was observed at 1, 3, and 7 days but not at 0 or 21 days after CSD. Semiquantitative lane analysis showed a maximal decrease of 90% for EAAT2 and 50% for EAAT1 at 3 days post-CSD. The neuronal isoform EAAT3 was unaffected by CSD. This period of down-regulation coincides with the time frame reported for induced ischemic tolerance. These data are consistent with reversal of glutamate transporter function contributing to glutamate release during ischemia and suggest that down-regulation of these transporters may contribute to ischemic tolerance induced by CSD.

ATP-Binding Cassette Transporters↗
PubMed2000

Culture models of neurodegenerative disease.

In order to investigate how mutant SOD1 protein or environmental exogenous stressors lead to the death of motor neurons, we have established several in vitro model systems. We describe some features of the various models in order to demonstrate the advantages and shortcomings of each system.

Animals↗
PubMed2000

[Selected CD34+ cells transplantation: a primary clinical report].

OBJECTIVE: CD34+ is an immunophenotype of hematopoietic stem cells/progenitors. CD34+ cells selection in vitro may deplete T-cells 4-5 logs and tumor cells 3-4 logs. It will benefit to mismatched related donor allo-transplantation and autologous transplantation of tumor diseases. METHODS: 19 patients aged 29 (5-52) years were treated by allo- (n = 5) or auto-PB CD34+ cells transplantation (n = 14). Grafts from fifteen patients with various disorders (3 lymphomas, 6 multiple myelomas, 2 SLE, 1 Sjogren's syndrome, 2 breast cancer and 1 medulloblastoma) and five haploidentical donors for leukemia patients (1 ALL-CR2, 2 AML-rel, 1 CML-CP and 1 MDS-RAEB) were isolated using magnetic-activated cell sorting (CliniMACS, Milteny Biotech, Germany). RESULTS: After separation, purity of CD34+ cells was > 97%. Depletion of CD34 negative cells was extensive: CD3+ 2.6-4.6 logs, CD4+ > 5 logs, CD8+ 4.6-> 5 logs, and CD19+ 1.2-3.1 logs. 19 patients who received selected CD34+ cells transplantation (CD34+ CT) were followed up for 11(1-20) months. Overall survival(OS) was 13/19 (68.4%) cases. 1/5 patients with haploidentical transplant had disease free survival for 19 months. 12/14(85.7%) auto-CD34+ CT achieved overall survival(OS). CONCLUSION: Selected CD34+ cells transplantation significantly decreases the incidence of > II GVHD and depletes tumor cell contamination. This approach may be useful to haploidentical or unrelated donor transplantation. It also benefits autologous transplant in various tumor disorders.

Adolescent↗
PubMed2000

Alternation of Na(+)-Ca2+ exchange in rat cardiac sarcolemmal membranes during different phases of sepsis.

OBJECTIVE: To study the alteration of Na(+)-Ca2+ exchange in rat cardiac sarcolemmal membrane during phases of septic shock. METHODS: Sepsis was induced by cecal ligation and puncture (CLP). Na(+)-Ca2+ exchange was assayed by radioactive analysis. RESULTS: Na(+)-dependent 45Ca2+ uptake was decreased by 62%-69% in late phase of sepsis, whereas it was not affected in early phase of sepsis. Na(+)-Ca2+ exchange stimulated by 5' guanylyl imidodiphosphate [Gpp (NH) p] was decreased by 65.7% in late phase of sepsis but unaltered in early phase of sepsis. Two agonists (angiotensin II and phenylephrine) coupled to Gq and a protein kinase C activator, phorbol 12-myristate 13-acetate (PMA) all inhibited Na(+)-Ca2+ exchange in late phase of sepsis. Na(+)-Ca2+ exchange activities induced by phosphorylation of Na(+)-Ca2+ exchange were decreased in late phase of sepsis, whereas inhibition of Na(+)-Ca2+ exchange by dephosphorylation was increased both in early and late phases of sepsis. CONCLUSION: The alteration of Na(+)-Ca2+ exchange during different phases of sepsis might be related to the activities of Gq, protein kinase C, and phosphorylation/dephosphorylation.

Animals↗
PubMed2000

[Alteration of interferon-gamma and interleukin-12 released by bronchoalveolar lavage cells from pulmonary tuberculosis].

OBJECTIVE: To explore the change of cytokine released in bronchoalveolar lavage fluid (BALF) cells from pulmonary tuberculosis (TB) patients. METHODS: BALF cells were obtained from group A: 10 normal control volunteers (normal control group); group B: 10 radiographically uninvolved sites of pulmonary TB patients (uninvolved sites group); group C: radiographically involved two lobes of lung and smear-positive of 10 pulmonary TB patients with an onset less than one year; and group D: radiographically involved more than three lobes and smear-positive of 10 pulmonary TB patients with an onset longer than two years. The IFN-gamma and IL-4 production in the culture supermate of BALF lymphocytes stimulated with PHA and IL-12 production in the culture supermate of BALF macrophage (AM) stimulated with lipopolysaccharide were determined using sandwich ELISA. RESULTS: In group C, the IFN-gamma and IL-12 levels [(834 +/- 101) and (294 +/- 73) ng/L, respectively] were not only significantly higher than those of the group B[(197 +/- 59) and (67 +/- 23) ng/L, respectively, all P < 0.01] and normal control group[(179 +/- 51) and (57 +/- 24) ng/L, respectively, all P < 0.01], but also significantly higher than those of group D[(431 +/- 169) and (121 +/- 63) ng/L, respectively, all P < 0.01]; in group D, the IFN-gamma and IL-12 levels were significantly higher than those of the group B and normal control group (all P < 0.01); there were no significant differences between the group C and normal control group (P > 0.05); no differences were seen in IL-4 level among the 4 study groups. CONCLUSIONS: A TH1 type response occurs in involved sites of pulmonary TB patients and that may result from AM which as a regulator through production of IL-12, and the TH1 type response may affect presentation and outcome in pulmonary TB.

Adult↗
PubMed1999

Mechanisms of transcriptional activation of bcl-2 gene expression by 17beta-estradiol in breast cancer cells.

bcl-2 gene expression is induced by 17beta-estradiol (E2) in T47D and MCF-7 human breast cancer cells, and the mechanism of E2 responsiveness was further investigated by analysis of the bcl-2 gene promoter. The -1602 to -1534 distal region (bcl-2j) of the promoter was E2-responsive; however, in gel mobility shift assays, the estrogen receptor alpha (ER(alpha)) did not bind [(32)P]bcl-2j, whereas Sp1 protein formed a retarded band complex. Further analysis demonstrated that the upstream region (-1603 to -1579) of the bcl-2 gene promoter contained two GC/GA-rich sites at -1601 (5'-GGGCTGG-3') and -1588 (3'-GGAGGG-5') that bound Sp1 protein. Subsequent studies confirmed that transactivation by E2 was dependent on ER(alpha)/Sp1 interactions with both GC-rich sites, and this was confirmed by in vitro footprinting. In contrast, a 21-base pair E2-responsive downstream region (-1578 to -1534) did not bind Sp1 or ER(alpha) protein; however, analysis of a complex binding pattern with nuclear extracts showed that ATF-1 and CREB-1 bound to this motif. These data coupled with results of transient transfection studies demonstrated that transcriptional activation by E2 of the -1578 to -1534 region of the bcl-2 gene promoter was dependent on induction of cAMP and subsequent activation through a cAMP response element. Thus, hormone regulation of bcl-2 gene expression in breast cancer cells involves multiple enhancer elements and E2-mediated transactivation does not require direct binding of the estrogen receptor with promoter DNA.

Activating Transcription Factor 1↗
PubMed1999

Cross-reactivity of myelin basic protein-specific T cells with multiple microbial peptides: experimental autoimmune encephalomyelitis induction in TCR transgenic mice.

Activation of autoreactive T cells is a crucial event in the pathogenesis of autoimmune diseases. Cross-reactivity between microbial and self Ags (molecular mimicry) is one hypothesis that could explain the activation of autoreactive T cells. We have systematically examined this hypothesis in experimental autoimmune encephalomyelitis using mice bearing exclusively myelin basic protein (MBP)-specific T cells (designated T+ alpha-). A peptide substitution analysis was performed in which each residue of the MBPAc1-11 peptide was exchanged by all 20 naturally occurring amino acids. This allowed the definition of the motif (supertope) that is recognized by the MBPAc1-11-specific T cells. The supertope was used to screen protein databases (SwissProt and TREMBL). By the search, 832 peptides of microbial origin were identified and synthesized. Of these, 61 peptides induced proliferation of the MBPAc1-11-specific transgenic T cells in vitro. Thus, the definition of a supertope by global amino acid substitution can identify multiple microbial mimic peptides that activate an encephalitogenic TCR. Peptides with only two native MBP-residues were sufficient to activate MBPAc1-11-specific T cells in vitro, and experimental autoimmune encephalomyelitis could be induced by immunizing mice with a mimic peptide with only four native MBP residues.

Amino Acid Substitution↗
PubMed1999

Differential expression of peroxiredoxin subtypes in human brain cell types.

The peroxiredoxin (Prx) protein is expressed widely in animal tissues and serves an antioxidant function associated with removal of cellular peroxides. We have cloned two Prx genes and observed differential expression of Prx-I and Prx-II (formerly NKEF-A and NKEF-B) in purified rat brain cell cultures (Sarafian et al. [1998] Mol. Chem. Neuropathol. 34:39-51). We have examined regional and cell-type-specific expression of Prx-I and Prx-II in paraffin sections of human brain using immunohistochemical methods. These studies revealed a clear segregation of expression of these two gene products in different brain cell types. In the cerebral cortex, cerebellum, basal ganglia, substantia nigra, and spinal cord, Prx-I was expressed primarily in astrocytes, while Prx-II was expressed exclusively in neurons. Prx-I was also prominently expressed in ependymal cells and subependymal matrix of substantia nigra and basal ganglia. Prx-II was not expressed at uniform density in all neurons. In general, small neurons such as cerebellar granule neurons displayed little or no staining, while large neurons, such as hippocampal pyramidal and Purkinje neurons were heavily stained. The absence of expression of Prx-I in neurons and the selective expression of Prx-II in large neurons suggest that these antioxidant enzymes serve distinct functional roles that may reflect the different functions and biochemical activities of these cell types. Restricted expression of these genes may also contribute to the selective vulnerability of these cells to a wide variety of neuropathologic conditions.

Adult↗
PubMed1999

Neural precursor cells differentiating in the absence of Rb exhibit delayed terminal mitosis and deregulated E2F 1 and 3 activity.

The severe neurological deficit in embryos carrying null mutations for the retinoblastoma (Rb) gene suggests that Rb plays a crucial role in neurogenesis. While developing neurons undergo apoptosis in vivo neural precursor cells cultured from Rb-deficient embryos appear to differentiate and survive. To determine whether Rb is an essential regulator of the intrinsic pathway modulating terminal mitosis we examined the terminal differentiation of primary cortical progenitor cells and bFGF-dependent neural stem cells derived from Rb-deficient mice. Although Rb -/- neural precursor cells are able to differentiate in vitro we show that these cells exhibit a significant delay in terminal mitosis relative to wild-type cells. Furthermore, Rb -/- cells surviving in vitro exhibit an upregulation of p107 that is found in complexes with E2F3. This suggests that p107 may partially compensate for the loss of Rb in neural precursor cells. Functional ablation of Rb family proteins by adenovirus-mediated delivery of an E1A N-terminal mutant results in apoptosis in Rb-deficient cells, consistent with the interpretation that other Rb family proteins may facilitate differentiation and survival. While p107 is upregulated and interacts with the putative Rb target E2F3 in neural precursor cells, our results indicate that it clearly cannot restore normal E2F regulation. Rb-deficient cells exhibit a significant enhancement of E2F 1 and 3 activity throughout differentiation concomitant with the aberrant expression of E2F-inducible genes. In these studies we show that Rb is essential for the regulation of E2F 1 and 3 activity as well as the onset of terminal mitosis in neural precursor cells.

Adenoviridae↗
PubMed1999

Prevention of neuronal cell death by neural adhesion molecules L1 and CHL1.

The effects of L1-Fc and CHL1-Fc fusion proteins on neuronal survival were investigated. Cerebellar granule neurons of mouse and hippocampal neurons of rat embryo undergo apoptosis when cultured in serum-free medium. Treatment with chimeric proteins containing the extracellular domains of the neural adhesion molecules L1 or CHL1 fused to the Fc region of human immunoglobulin significantly enhanced the survival of neurons. Compared to the control, the percentage of surviving neurons increased about 60% and 45% with L1 and CHL1 fusion proteins, respectively. A fusion protein containing the extracellular domain of NCAM had no effect on survival. The L1 and CHL1 fusion proteins were effective both in soluble form or when offered as a substrate, with the maximal effect at about 1 microg/mL. To explore the intracellular events related to the neuronal survival effects of L1-Fc fusion protein, Bcl-2 and c-Jun expression were analyzed by Western blotting. The level of Bcl-2 in cerebellar granule neurons was increased by treatment with L1-Fc at both 1 and 5 days of culture. The level of c-Jun was not significantly affected at the early time point and was reduced by L1-Fc fusion protein after long-term culture. The results demonstrate that the neural adhesion molecule L1 and its relative CHL1 are potential neuronal survival factors for neurons of the central nervous system. Bcl-2 may serve as one of the intracellular mediators of the neuronal survival effects of L1.

Animals↗