Search PubMed⌕ Search

Biomedical subjects

L Zhao

Publications and source records attributed to L Zhao.

At least 19 recordsLinked to original sources
PubMed1996

RAGE and amyloid-beta peptide neurotoxicity in Alzheimer's disease.

Amyloid-beta peptide is central to the pathology of Alzheimer's disease, because it is neurotoxic--directly by inducing oxidant stress, and indirectly by activating microglia. A specific cell-surface acceptor site that could focus its effects on target cells has been postulated but not identified. Here we present evidence that the 'receptor for advanced glycation end products' (RAGE) is such a receptor, and that it mediates effects of the peptide on neurons and microglia. Increased expressing of RAGE in Alzheimer's disease brain indicates that it is relevant to the pathogenesis of neuronal dysfunction and death.

Alzheimer Disease↗
PubMed1996

Orientation of paramagnetic probes attached to gizzard regulatory light chain bound to myosin heads in rabbit skeletal muscle.

The orientation of the myosin neck was monitored using electron paramagnetic resonance (EPR) spectroscopy. Gizzard regulatory light chain was labeled with a nitroxide spin probe and exchanged for the native subunit, located in the myosin neck, in rabbit psoas muscle fibers. The EPR spectra of rigor fibers indicated a substantial degree of probe immobilization and showed a strong dependence on the orientation of the fiber axis relative to the magnetic field, indicating that the neck was ordered in this state. Spectra of relaxed fibers at 24 degrees C showed that the neck was disordered, but the spectra of relaxed fibers at 4 degrees C indicated that the neck was partially ordered. Active fibers at the two temperatures produced spectra identical to relaxed fibers, indicating that no novel angles could be seen in the neck during the powerstroke. Proteolytic fragments of myosin, S1 and HMM, were exchanged with labeled light chains and bound to thin filaments in unlabeled fibers. The distribution of probe orientations for HMM was identical to that of labeled rigor fibers, while S1 showed a slightly different distribution, suggesting that the neck is distorted (by a few degrees) by the interactions of the two heads of myosin when bound to actin.

Animals↗
PubMed1996

Regulation of cytokine mRNA expression in activated lymphocytes by human choriocarcinoma JAR cells.

Because the fetus is semiallogenic to the mother, considerable modulation of the maternal immune response must occur in order for pregnancy to be successfully carried to term. Some authors have hypothesized that the immunomodulation of pregnancy includes an adjustment of cytokine responses away from the Th1 paradigm and toward the Th2 pattern. In vivo data from murine pregnancy support this hypothesis. However, in humans, the Th1/Th2 model appears to be more complex than that in mice, and cytokine expression of mRNA in human decidual tissue does not reflect a clear-cut Th2 bias. The experiments described here were undertaken to determine whether and how trophoblastic cells modulate cytokine expression in activated lymphocytes, and whether there is a trend toward the use of the Th2 pattern in an experimental model of the maternal-fetal interface. We used reverse transcriptase polymerase chain reaction (rtPCR) to detect cytokine mRNA expression in human peripheral blood mononuclear cells cocultivated with human choriocarcinoma JAR cells. We found that although IL-2 (a paradigmatic Th1 cytokine) was significantly down-regulated by JAR cells at the mRNA level, similar decreases were also seen in IL-10, which participates in the Th2 paradigm. We were unable to detect changes in either interferon-gamma (IFN-gamma, a Th1 cytokine) or IL-4 (a Th2 cytokine) mRNA's or in IL-2R expression by fluorescence-activated cell sorting. These studies indicate that human choriocarcinoma JAR cells are capable of modifying cytokines in activated lymphocytes other than those involved in the Th1 paradigm. While it may be useful to view human responses against the background of these patterns established from murine systems, it is reasonable to conclude that human pregnancy may not involve regulation of Th1 immune responses exclusively.

Base Sequence↗
PubMed1996

Lysis of adult ventricular myocytes by cells infiltrating rejecting murine cardiac allografts.

BACKGROUND: Immunologic mechanisms that mediate myocardial cell injury during rejection are not fully understood. We therefore investigated whether cells that infiltrate rejecting cardiac allografts are capable of directly injuring myocytes and whether this injury resembles that produced by cytotoxic T lymphocytes (CTLs) that are generated in a mixed lymphocyte reaction (MLR). METHODS AND RESULTS: Heart-infiltrating cells (HICs) were isolated from murine heterotopic BALB/c cardiac allografts undergoing rejection 6 to 8 days after transplantation into C57BL/6 mice. An in vitro model system of cultured adult murine ventricular myocytes was developed to facilitate investigation of cell-mediated myocyte injury. Isolated adult myocytes were incubated with either HICs or MLR effector cells, and myocyte death was quantified by counting the number of rod-shaped myocytes excluding trypan blue. The frequency of donor-reactive CTLs was similar in the HIC and MLR populations, as assessed by limiting dilution analysis. However, HICs were less efficient at killing donor-strain myocytes than were MLR cells. CTL-mediated cell lysis occurred by 6 hours, whereas myocyte injury produced by HICs was more gradual, with considerable cytotoxicity occurring between 12 and 24 hours. Furthermore, whereas MLR cells lysed only donor-strain myocytes, HIC lysed donor, third-party, and syngeneic myocytes. Treatment of MLR cells and HICs with anti-CD8 antibody plus complement produced a much greater inhibition of MLR cytotoxicity than of HIC cytotoxicity. CONCLUSIONS: These data demonstrate that only a small component of myocyte injury mediated by allograft-infiltrating cells can be ascribed to CTLs within the infiltrating cell population. These findings suggest that cell types associated with a delayed-type hypersensitivity response, as well as CTLs, cause myocyte injury during cardiac rejection.

Animals↗
PubMed1996

Preservation of global cardiac function in the rabbit following protracted ischemia/reperfusion using monophosphoryl lipid A (MLA).

Monophosphoryl lipid A (MLA), a derivative of the minimal substructure of lipopolysaccharide (lipid A) possesses immunomodulatory activity of the parent lipid A yet enjoys reduced toxicity. It has previously been reported that pretreatment with MLA reduces myocardial infarct size and stunning in dogs following ischemia and reperfusion. The aim of this study was to evaluate the ability of monophosphoryl lipid A (MLA) to preserve global cardiac function and peripheral hemodynamics in a rabbit model of prolonged regional ischemia (90 min), and reperfusion (6 h). An evaluation of potential mechanisms by which MLA may preserve cardiac function was also undertaken. Single dose pretreatment with MLA (35 micrograms/kg i.v.) 24 h prior to ischemia resulted in significant improvement in left ventricular developed pressure, dP/dt, rate-pressure product and mean arterial pressure during reperfusion (P < 0.05 v control). Although in this model of prolonged ischemia MLA pretreatment did not reduce infarct size (54.5 +/- 11.4% in control v 63.3 +/- 8.3% in MLA, P = N.S.), evaluation of myocardial adenylate and adenosine catabolite pools at the end of ischemia indicated a preservation of ATP and ADP and a decreased production of downstream adenosine catabolites including inosine, xanthine and uric acid. Adenosine kinase, but not 5'-nucleotidase (5'-NTase) or adenosine deaminase activity determined following reperfusion was 76% and 60% higher (P < 0.05) in non-risk and post-ischemic myocardium of MLA pretreated rabbits compared with controls. Although there was a trend toward lower tissue myeloperoxidase activity in post-ischemic myocardium from treated rabbits, the results were not significantly different from control animals. These results suggest that a 24-h pretreatment with MLA, without further treatment during ischemia or reperfusion was associated with: (1) preservation of global myocardial function during reperfusion; (2) preservation of myocardial high energy adenylates and reduced formation of adenosine catabolites during ischemia; (3) elevated myocardial adenosine kinase activity. Increased recycling of adenosine to phosphorylated nucleotides may result from MLA's affect on adenosine kinase, which could explain the drugs effect on adenylate and adenosine metabolite pools.

5'-Nucleotidase↗
PubMed1996

Effects of epidermal growth factor (EGF) and colony-stimulating factor-1 (CSF-1) on expression of c-fos in rat mandibular molars: implications for tooth eruption.

The c-fos gene is expressed in the dental follicle of the first mandibular molar of rats. Molecules that accelerate tooth eruption, epidermal growth factor (EGF) and colony-stimulating factor-1 (CSF-1), enhance the expression of the c-fos gene in vitro in both a time- and concentration-dependent manner. In vivo, EGF enhances c-fos expression in the follicle from day 0-7 postnatally, but by day 9 the follicle is refractory to this stimulus. Immunostaining reveals that the c-fos gene is translated in the cultured dental follicle cells, with staining seen in the nucleus and the perinuclear region of the cytoplasm. In vivo, immunostaining for c-fos is prominent in the dental follicle early postnatally, with little or no staining seen in the stellate reticulum and dental pulp. By day 10 postnatally, staining for c-fos is greatly reduced in the dental follicle. Thus, the presence of c-fos early postnatally in the tissue required for eruption, the dental follicle, as well as the enhancement of c-fos gene expression in the follicle by EGF or CSF-1, suggests that c-fos plays a role in tooth eruption. That role may be either to promote differentiation of mononuclear cells into osteoclasts needed for eruption or to recruit the mononuclear cells into the follicle.

Animals↗
PubMed1996

Systemic uptake and clearance of chloroform by hairless rats following dermal exposure. I. Brief exposure to aqueous solutions.

The systemic uptake of chloroform from dilute aqueous solutions into live hairless rats under conditions simulating dermal environmental exposure was studied. Whole blood was sampled during a 30-min immersion of an animal within water containing a known concentration of chloroform and then for 5.5 h following its removal from the bath. The amount of chloroform systemically absorbed was determined by comparing the AUCs of the blood concentration vs. time plots from dermal exposure to that obtained after i.v. infusion (for a period of 30 min) of an aqueous solution containing a known amount of chloroform (positive control). Although dermal data implied two-compartment disposition characteristics, i.v. infusion data fit best to a three-compartment disposition. Linear pharmacokinetics was observed both by i.v. administration and percutaneous absorption at the dose levels studied. Chloroform was detected in the rat blood as early as 4 min following exposure. Our findings suggest that about 10.2 mg of chloroform was systemically absorbed after dermal exposure of a rat to an aqueous solution of 0.44 mg/ml. This amount is substantially higher than the predictions of mathematical risk-models put forth by some investigators. However, when expressed as the "effective" permeability coefficient (Kpeff), close agreement was noticed between our value and those estimated by others using physiologically based pharmacokinetic (PBPK) models. Also, in terms of Kpeff, reasonable agreement existed between our and another investigator's past estimates of uptake based on depletion of bath level of chloroform and the actual uptake measured in our current experiments. The estimated onset of systemic entry seen here is entirely consistent with our estimate of how long it takes to establish the diffusion gradient across the stratum corneum based on tape stripping.

Animals↗
PubMed1996

Erwinia amylovora secretes harpin via a type III pathway and contains a homolog of yopN of Yersinia spp.

Type III secretion functions in flagellar biosynthesis and in export of virulence factors from several animal pathogens, and for plant pathogens, it has been shown to be involved in the export of elicitors of the hypersensitive reaction. Typified by the Yop delivery system of Yersinia spp., type III secretion is sec independent and requires multiple components. Sequence analysis of an 11.5-kb region of the hrp gene cluster of Erwinia amylovora containing hrpI, a previously characterized type III gene, revealed a group of eight or more type III genes corresponding to the virB or lcrB (yscN-to-yscU) locus of Yersinia spp. A homolog of another Yop secretion gene, yscD, was found between hrpI and this group downstream. Immediately upstream of hrpI, a homolog of yopN was discovered. yopN is a putative sensor involved in host-cell-contact-triggered expression and transfer of protein, e.g., YopE, to the host cytoplasm. In-frame deletion mutagenesis of one of the type III genes, designated hrcT, was nonpolar and resulted in a Hrp- strain that produced but did not secrete harpin, an elicitor of the hypersensitive reaction that is also required for pathogenesis. Cladistic analysis of the HrpI (herein renamed HrcV) or LcrD protein family revealed two distinct groups for plant pathogens. The Yersinia protein grouped more closely with the plant pathogen homologs than with homologs from other animal pathogens; flagellar biosynthesis proteins grouped distinctly. A possible evolutionary history of type III secretion is presented, and the potential significance of the similarity between the harpin and Yop export systems is discussed, particularly with respect to a potential role for the YopN homolog in pathogenesis of plants.

Amino Acid Sequence↗
PubMed1996

Adrenomedullin activity in chronically hypoxic rat lungs.

Adrenomedullin (AM) is a novel vasodilator with structural similarities to calcitonin gene-related peptide (CGRP). This study investigated AM activity in the rat lung during hypoxia-induced pulmonary hypertension. Both rat AM (0.2-10 nmol) and alpha-CGRP (0.2-2 nmol) produced dose-related reductions in pulmonary artery pressure in the isolated perfused lung ventilated with 2% O2. Pretreatment with alpha-CGRP, which demonstrated tachyphylaxis, or its antagonist, CGRP-(8-37), reduced the hypotensive response to AM, suggesting that part of the response to AM is mediated by CGRP receptors. 125I-labeled AM and 125I-labeled CGRP binding was significantly increased in lung membranes from 7-day hypoxic animals (AM from 1.94 +/- 0.3 to 3.36 +/- 0.4 and CGRP from 0.06 +/- 0.01 to 0.12 +/- 0.02 pmol/mg protein), with no change in dissociation constant. Moreover, the hypotensive response to both peptides was increased in the lungs of 7-day hypoxic rats. There was no significant change in lung immunoreactive AM concentrations (hypoxic 5.04 +/- 0.48 vs. control 6.28 +/- 0.76 pmol/g wet wt of tissue) or steady-state AM mRNA levels in 7-day hypoxic rats. Nonetheless, AM may be useful for the acute pharmacological manipulation of pulmonary artery pressure in hypoxia-induced pulmonary hypertension.

Adrenomedullin↗
PubMed1996

Sequestration of inhaled particulate antigens by lung phagocytes. A mechanism for the effective inhibition of pulmonary cell-mediated immunity.

Dendritic cells (DCs) have emerged as the dominant antigen-presenting cells (APCs) of the lung, playing a vital role in the induction of cell-mediated immunity to inhaled antigens. We have previously demonstrated that an airway challenge with the soluble antigen hen egg lysozyme yields rapid acquisition of specific antigen-presenting cell activity by purified pulmonary DCs and a cell-mediated immune response in the lung upon secondary challenge. To examine how a particulate antigen leads to a cell-mediated response in vivo, graded concentrations of heat-killed Listeria (HKL) were injected intratracheally into Lewis rats. The bacteria were rapidly ingested by lung macrophages and polymorphonuclear leukocytes. The ability of purified pulmonary DCs pulsed in vivo by an airway challenge with HKL to subsequently stimulate HKL-specific responses ex vivo showed a threshold response, requiring a dose in excess of 10(9) organisms/rat. By contrast, all dosages of HKL yielded specific sensitization of lymphocytes in the draining bilar nodes. Pulmonary DCs purified from rats after a secondary in vivo airway challenge with HKL at day 14 were ineffective antigen-presenting cells except at high dosages of antigen. The generation of cell-mediated pulmonary inflammation paralleled the antigen-presenting cell activity of pulmonary DCs and was observed only at high antigen dosages. Hen egg lysozyme immobilized onto polystyrene beads and injected intratracheally yielded comparable results to those observed with HKL. We suggest that a pulmonary cellular immune response is generated to an inhaled particulate antigen when the protective phagocytic capacities of the lung are exceeded and antigen is able to interact directly with interstitial DCs. The diversion of particulate antigens by pulmonary phagocytes may help to limit undesirable pulmonary inflammation while allowing the generation of antigen-specific immune lymphocytes in vivo.

Animals↗
PubMed1996

Evaluation of the redox tolerance index on hepatic energy reserve of hepatitis B patients.

OBJECTIVE: To detect the redox tolerance index (RTI) for hepatic energy reserve. PATIENTS AND METHODS: 27 patients with chronic active hepatitis B (CAH) and 34 patients with post-hepatitis B liver cirrhosis (LC) were studied. RESULTS: The RTI was significantly lower in CAH and LC cases than in normal controls (P < 0.01). There was no difference between CAH and LC. The RTI of decompensated LC was significantly lower than that of the compensated LC (P < 0.05), indicating that there was no correlation between RTI and the parameters of routine liver functions. CONCLUSION: It is suggested that the RTI based on redox theory is effective and accurate in predicting hepatic energy reserve.

Adult↗
PubMed1996

[Ion-selective microelectrodes: principle and application of in vivo measurements of ionic concentrations in cochlear endolymph].

OBJECTIVE: To introduce the method of making neutral carrier ion-selective microelectrode and use it for in vivo measurement of ionic concentrations in cochloear endolymph. METHODS: GG-17 glass capillaries with 1.85 mm O.D. were rinsed extensively and dried in an oven. Two capillaries were parallelly connected and pulled with microelectrode puller to make a double-barreled micropipette. One barrel was back-filled with ion exchanger and internal reference solution, which served as ionic potential electrode, while the other barrel filled with 150 mmol/L KCL as reference electrode. Each barrel of microelectrode was connected to differential electrometer via Ag-AgCl wire and output was recorded on a three-channel recorder. Every ion-selective microelectrode was calibrated in a series of standard solutions to determine the required characteristics. Twenty healthy guinea pigs with normal hearing were anesthetized and were artificially respired through the tracheal canal after the intramuscular injection of suxamethonium chloride. The tympanic bulla was exposed and a double-barreled ion-selective electrode was inserted into the scala media through the round window and basilar membrane. The ionic potentials and endocochlear potentials (EP) were simultaneously recorded from the basal turn of the cochlea. Ionic concentrations were then calculated by Nicolsky-Eisenman equation. RESULTS: The concentrations of potassium, sodium and calcium ions in the cochlear endolymph were 146.3 +/- 11.8 mmol/L, 0.36 +/- 0.22 mmol/L and 16.2 +/- 5.7 micromol/L, respectively. CONCLUSIONS: The neutral carrier ion-selective microelectrode made with the above method is reliable and therefore can be used for accurately measuring the ionic concentrations of microenvironment in vivo continuously and transiently.

Animals↗
PubMed1996

[Histopathological and immunohistochemical studies on gliosarcoma].

Immunohistochemical staining of 5 gliosarcomas was performed. Neoplastic glial component stained positive by GFAP. Endothelial cells lining the lumina of glomeruloid vascular structures stained positively with both UEA-1 and FVIII/RAg antibodies. Mesenchymal cells of sarcomatous areas stained positively with SMSA antibody and presence of PDGFR. The staining results demonstrate that the sarcomatous component of gliosarcoma is of smooth muscle origin and suggest that the vascular smooth muscle hyperplasia is related to PDGF.

Aged↗
PubMed1996

Nitrite, N-nitroso compounds, and other analytes in physiological fluids in relation to precancerous gastric lesions.

Levels of gastric juice nitrite, several urinary N-nitroso compounds, and other analytes were examined among nearly 600 residents in an area of Shandong, China, where precancerous gastric lesions are common and rates of stomach cancer are among the world's highest. Gastric juice nitrite levels were considerably higher among those with gastric juice pH values above 2.4 versus below 2.4. Nitrite was detected more often and at higher levels among persons with later stage gastric lesions, especially when gastric pH was high. Of those with intestinal metaplasia, 17.5% had detectable levels of gastric nitrite, while this analyte was detected in only 7.2% of those with less advanced lesions. Relative to those with undetectable nitrite, the odds of intestinal metaplasia increased from 1.5 (95% confidence interval = 0.6-4.1) to 4.1 (95% confidence interval = 1.8-9.3) among those with low and high nitrite concentrations, respectively. Urinary acetaldehyde and formaldehyde levels also tended to be higher among those with more advanced pathology, particularly dysplasia. However, urinary excretion levels of total N-nitroso compounds and several nitrosamino acids differed little among those with chronic atrophic gastritis and intestinal metaplasia and dysplasia, consistent with findings from recent studies in the United Kingdom, France, and Colombia. The data from this high-risk population suggest that elevated levels of gastric nitrite, especially in a high pH environment, are associated with advanced precancerous gastric lesions, although specific N-nitroso compounds were not implicated.

Adult↗
PubMed1995

Structure of the gene for the catalytic subunit of human DNA polymerase delta (POLD1).

We have isolated genomic DNA clones covering the gene for human DNA polymerase delta catalytic subunit (POLD1) and its 5' flanking sequence. This gene is divided into 27 exons and is distributed over at least 32 kb of DNA. The exons and most of the introns are relatively small. The sizes of the exons range from 55 to 201 bp. Seven introns are smaller than 100 bp. Intron 1 is the largest intron, with a size of greater than 10 kb. All of the intron-exon junctions match well with the reported consensus sequences. Multiple copies of the Alu repetitive sequence and the variable number of tandem repeats were found in several introns. Transcription of POLD1 appears to initiate at multiple sites. The major start site was 53 nucleotides upstream of the ATG start codon. The sequence of the promoter and upstream DNA is G+C rich and does not contain a TATA sequence. Several potential transcription factor-binding sites, including the AP2-, CTF-, Ets1-, GCF-, MBF-1-, NF-E1-, and Sp1-binding sites, were found in this region. A 1.8-kb pol delta promoter DNA directed the expression of a luciferase reporter gene when transfected into HeLa cells.

Base Sequence↗
PubMed1995

Detection of residual leukemic cells in patients with acute promyelocytic leukemia by the fluorescence in situ hybridization method: potential for predicting relapse.

The translocation between chromosomes 15 and 17, t(15;17)(q22-24;q11-21), is present in the bone marrow cells of most patients with acute promyelocytic leukemia (APL). Although conventional cytogenetic methods are useful for diagnosing this disease, difficulties are experienced in detecting residual disease among those patients who have achieved remission. In this study, we used the fluorescence in situ hybridization (FISH) method to attempt to detect residual leukemic cells in 10 APL patients in clinical remission. The duration of remission ranged from 2 to 93 months at the time of study. Multiple bone marrow samples were analyzed by FISH in most patients. In 6 patients, no cell with t(15;17) was found. These patients remain in complete remission at present (approximately 25 to 33 months since first studied by FISH). In 4 patients, low frequencies of cells with t(15;17) were observed in at least one bone marrow sample examined. All of these patients relapsed within 1 to 14 months. No cell with t(15;17) was identified by the conventional G-banding method in any sample. The FISH results correlated well with that of a two-round nested reverse transcription polymerase chain reaction assay that was performed on the same samples. Thus, our study suggests that FISH is potentially a useful tool for detecting residual APL cells and for identifying patients at high risk of relapse.

Base Sequence↗
PubMed1995

Separation of pure populations of epithelial cells from rabbit distal colon.

A simple method using divalent chelators is described for the isolation of viable populations of surface and crypt cells from rabbit distal colon. Histological studies were performed to monitor colonocyte dissociation and determine contamination by nonepithelial cells. Cell viability was assessed by trypan blue exclusion assay and by 22Na uptake measurements. Electron microscopy was used to determine the integrity of the isolated cells. Alkaline phosphatase and [3H]thymidine uptake were measured to assess the purity of the different cell fractions. Combined fractions 4 and 5 contained the highest percentage of pure surface cells, while fractions 10, 11, and 12 were predominantly crypts. Alkaline phosphatase activity was 13 +/- 3-fold higher in the surface cells than in the crypt cells, while [3H]thymidine uptake was 8 +/- 4-fold higher in the crypt cells than in the surface cells. Amiloride-sensitive and -insensitive 22Na uptake was the same in the surface cells directly after isolation and after 3 h in culture. In this study we demonstrate a method for the preparation of highly enriched fractions of rabbit colon surface and crypt cells that remain viable and functional in short-term culture.

Animals↗