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Biomedical subjects

G Lin

Publications and source records attributed to G Lin.

At least 145 records · Page 8Linked to original sources
PubMed1995

Effects of interleukin-1 and interleukin-1 receptor antagonist in AIDS-Kaposi's sarcoma.

Kaposi's sarcoma (KS) is the most common tumor seen in patients with HIV-1 infection. HIV-1 may induce KS directly through viral protein(s) or indirectly through regulation of cytokines such as IL-1 and IL-6. We have shown that AIDS-KS spindle cells express IL-1 beta and that IL-1ra inhibits KS-spindle cell growth. IL-1ra had little effect on human umbilical vein endothelial cells (HUVEC), human aortic smooth muscle cells (HASM), and human foreskin fibroblast (NN41). These findings support an autocrine activity for IL-1. Furthermore, exogenous IL-1 can enhance AIDS-KS cell growth, and this effect is completely blocked by IL-1ra. As expected, IL-1ra also blocks IL-1 mediated upregulation of IL-6 and bFGF, both of which are autocrine growth factors for KS. IL-1ra is thus a potential candidate for the treatment of AIDS-associated KS.

Acquired Immunodeficiency Syndrome↗
PubMed1995

Interleukin-4 suppresses the expression of macrophage NADPH oxidase heavy chain subunit (gp91-phox).

The production of superoxide anion by NADPH oxidase is a principal nonspecific bactericidal activity of macrophages and neutrophils in host defense. However, exuberant production of superoxide anion also damages host tissues. Cloning and DNA sequencing of the 91 kDa subunit (gp91-phox) open reading frame indicated a high degree of sequence conservation, greater than 90% in nucleotide and amino acid sequences, between the porcine and human cDNAs. We show in pigs that interleukin-4 (IL-4), a T lymphocyte cytokine which plays a major role in mediating antibody responses to pathogens, suppresses superoxide anion production in macrophages by specifically reducing the level of mRNA encoding gp91-phox. Messenger RNA levels are suppressed approx. 70% within 4 h and persist for 24 h without any change in the rate of mRNA turnover. Nuclear run-on analysis showed that IL-4 did not alter the rate of gp91-phox gene transcription under conditions in which IL-1 beta transcription was inhibited. These results indicate that IL-4 suppresses the inflammatory response of macrophages by mechanisms that include post-transcriptional regulation of the 91 kDa catalytic subunit of NADPH oxidase, and transcriptional regulation of inflammatory cytokine expression.

Animals↗
PubMed1995

Expression of CD34 in endothelial cells, hematopoietic progenitors and nervous cells in fetal and adult mouse tissues.

The human cell surface molecule CD34 is selectively expressed on uncommitted and committed hematopoietic progenitor cells and on vascular endothelial cells. It has been suggested that CD34 regulates early events in blood cell migration and differentiation, possibly as a cell adhesion molecule. To characterize the patterns of expression of CD34 in the mouse embryo and in the adult, as well as to dissect the function of different portions of the extracellular domain of this molecule, we have generated the first monoclonal antibodies (mAb) specific for mouse CD34. The epitope(s) recognized by these mAb are not carbohydrate moieties, and are comprised either within the immunoglobulin-like domain or within a portion of the mucin domain, containing approximately half of the predicted O- and N-linked carbohydrate attachment sites. The specificity of the antibodies was established by ELISA and Western blotting. Western analysis revealed that these mAb recognize a protein of approximately 110 kDa in PA6 stromal cell lysates, which can be specifically blocked by the recombinant CD34 protein. To establish the reactivity of these mAb on different cell lineages, a panel of cell lines was stained. This analysis showed strong reactivities with 3T3 fibroblasts, stromal cell lines from fetal liver and with the endothelial cell line D10. Bone marrow hematopoietic progenitors were also stained by these mAb. Immunostaining of frozen sections from embryonic and adult tissues revealed a strong reactivity against vascular endothelial cells at different stages of development, including sinusoidal cells in the fetal liver, yolk sac, and in the fetal bone marrow, endothelial cells from adult lung and kidney, and neural cells, including those of the neural tube of midgestation embryos and neuronal bodies in adult brain.

3T3 Cells↗
PubMed1995

Cloning, expression, and purification of an anti-desipramine single chain antibody in NS/O myeloma cells.

Drug-specific monoclonal antibodies and their antigen-binding Fab fragments reverse acute desipramine toxicity in a rat experimental model by inducing a redistribution of drug from cardiac tissue into serum and extracellular fluid. In order to investigate the use of smaller recombinant antibody fragments such as single chain Fv (sFv) as an antidote, an efficient murine NS/O myeloma expression system was developed. The variable light (VL) and variable heavy (VH) domains of a murine anti-desipramine monoclonal antibody were cloned and sequenced. A 270 amino acid VH-(Gly4Ser)3-VL sFv was prepared by overlapping polymerase chain reaction (PCR) amplification of VH with heavy chain leader peptide, VL, and the linker. This construct was subcloned into a mammalian expression vector which utilizes the SR alpha promoter, a hybrid promoter consisting of the SV40 early promoter with portions of the human T-cell leukemia virus type I long terminal repeat and also containing the Escherichia cloi xanthine-guanine phosphoribosyltransferase gene for selection. NS/O myeloma cells were transfected by electroporation. Stable recombinant NS/O clones were screened for expression of sFv using reverse transcriptase-PCR to detect mRNA and an enzyme-linked immunosorbent assay (ELISA) to detect sFv. Secreted sFv from clones capable of growth to a cell density of 2-4 x 10(6) viable cells/mL was purified in a single step using a desipramine affinity column resulting in 12-39 mg/L of purified sFv. Affinity-purified sFv had comparable desipramine binding activity to Fab when evaluated by competitive ELISA.

Amino Acid Sequence↗
PubMed1995

Comparison of serum C-reactive protein concentrations for laparoscopic versus open cholecystectomy.

In an attempt to quantify the difference in tissue damage between open cholecystectomy (OC) and laparoscopic cholecystectomy (LC), we have compared in a prospective manner the pre- and post-operative concentrations of serum C-reactive protein (CRP) in 17 patients undergoing LC and 13 patients undergoing OC. In addition, we measured the pre- and postoperative white blood cell counts (WBC), the postoperative body temperature, and the postoperative duration of hospitalization. There were no differences in the preoperative serum CRP concentrations--5.9 +/- 2.62 mg/l (mean +/- SD) for the LC group and 6.12 +/- 2.38 mg/l for the OC group. Serum CRP rose markedly following OC compared to that of patients who underwent LC (128.6 +/- 45.1 mg/l vs 26.8 +/- 10.5 mg/l) (P < 0.001). There were also significant differences in the postoperative WBC count (14,000 +/- 2,900 cells for the OC group vs 10,600 +/- 3,000 cells for the LC group), the postoperative body temperature (37.5 +/- 0.3 degrees C vs 37.0 +/- 0.3 degrees C), and the postoperative hospital stay (5.5 +/- 1.5 days vs 1.9 +/- 0.9 days). There was no correlation between serum CRP concentrations and the other postoperative parameters. These results provide us with biochemical evidence supporting the clinical observation that LC is far less traumatic to the patient than OC.

Adult↗
PubMed1995

The fate of long-standing intraperitoneal gallstone in the rat.

Gallstones are occasionally spilled into the peritoneal cavity during open and laparoscopic cholecystectomy. Using the rat model, we investigated the long term effect of such retained intraperitoneal gallstones. During a follow-up period of one year, no systemic deleterious outcome could be attributed to the presence of the implanted gallstones except for mild local effects. Based on the rat model we conclude that reasonable effort should be made in order to retrieve an escaped gallstone, but there is no justification whatsoever for a conversion of the laparoscopic procedure to an open laparotomy only for the purpose of retrieving a lost stone.

Animals↗
PubMed1995

Puqiedinone, a novel 5 alpha-cevanine alkaloid from the bulbs of Fritillaria puqiensis, an antitussive traditional Chinese medicine.

A novel 5 alpha-cevanine alkaloid, puqiedinone [1], was isolated from the bulbs of Fritillaria puqiensis, a traditional Chinese medicine used widely for its antitussive and expectorant properties. The structure of 1 was assigned as (20R,22S,25R)-20-deoxy-5 alpha-cevanine-3 beta-ol-6-one based on spectral analysis and comparison with the structures of related known compounds. Puqietinone [2] was also identified from the same herbal plant and the structure was revised as (22R,25S)-N-methyl-22,26-epiminocholest-3 beta-ol-6-one based on spectroscopic and X-ray crystallographic methods.

Antitussive Agents↗
PubMed1995

Induction of inflammatory cytokines in bovine alveolar macrophages following stimulation with Pasteurella haemolytica lipopolysaccharide.

Bovine tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) cDNAs were generated by reverse transcription and then by PCR amplification from lipopolysaccharide (LPS)-stimulated alveolar macrophage RNA. The amplified cDNAs were cloned into pPow and expressed in Escherichia coli DH5 alpha. The expressed proteins were confirmed as TNF-alpha and IL-1 beta by Western blot (immunoblot) analysis and bioassays. We then used the cloned genes as probes in Northern (RNA) blots and investigated the kinetics of TNF-alpha and IL-1 beta mRNA expression in bovine alveolar macrophages stimulated with purified LPS from Pasteurella haemolytica 12296. The effect of LPS on TNF-alpha and IL-1 beta gene expression was dose dependent, and induction was observed at a concentration of 0.01 microgram/ml. Both TNF-alpha and IL-1 beta mRNA expression were detectable within 0.5 h after stimulation with 1 microgram of LPS per ml, peaked at 1 to 2 h, steadily declined up to 16 h, and were undetectable by 24 h. Secreted TNF-alpha measured by bioassay peaked at 4 h and accumulated at a lesser concentration in conditioned medium throughout the 24 h. By contrast, secreted IL-1 beta was induced at 8 h and reached a maximal concentration at 24 h after stimulation. The ability of LPS to induce TNF-alpha and IL-1 beta gene expression and secretion of bioactive proteins were suppressed by polymyxin B. Our findings support a role for LPS from P. haemolytica in the induction of inflammatory cytokines in bovine pneumonic pasteurellosis.

Animals↗
PubMed1995

Effects of pesticides on the ratio of 16 alpha/2-hydroxyestrone: a biologic marker of breast cancer risk.

Xenobiotic estrogens are external compounds with estrogenic activity that may thereby affect the risk of breast cancer. This paper describes a mechanism by which xeno-estrogens may affect the development of breast cancer. Estradiol metabolism proceeds by hydroxylation at one of two mutually exclusive sites at C-2 and C-16 alpha. The catechol pathway yields the weakly estrogenic 2-hydroxyestrone (2-OHE1), which inhibits breast cell proliferation. In contrast, the alternative pathway yields the genotoxic 16 alpha-hydroxyestrone (16 alpha-OHE1), which enhances breast cell growth, increases unscheduled DNA synthesis, and oncogene and virus expression, and increases anchorage-independent growth. Using a radiometric assay that measures the relative formation of 16 alpha-OHE1 versus 2-OHE1 from specifically tritiated estradiol in (ER+) MCF-7 cells, we compared the ratio of 16 alpha-OHE1/2-OHE1 observed after treatment with the known rodent carcinogen 7,12-dimethylbenz[a]anthracene (DMBA) with the ratios after treatment with DDT, atrazine, gamma-benzene hexachloride, kepone, coplanar PCBs, endosulfans I and II, linoleic and eicosapentenoic acids, and indole-3-carbinol (I3C). These pesticides significantly increase the ratio of 16 alpha-OHE1/2-OHE1 metabolites to values comparable to or greater than those observed after DMBA. In contrast, the antitumor agent I3C increased 2-OHE1 formation and yielded ratios that are 1/3 of those found in unexposed control cells and 1/10th of those found in DMBA-treated cells. Thus the ratio of 16 alpha-OHE1/2-OHE1 may provide a marker for the risk of breast cancer. Assays of this ratio, which can be measured in spot urines, may prove useful for a variety of in vitro and in vivo studies bearing on breast cancer risk.

Biomarkers↗
PubMed1994

Structures of human and porcine aldehyde reductase: an enzyme implicated in diabetic complications.

The crystal structures of porcine and human aldehyde reductase, an enzyme implicated in complications of diabetes, have been determined by X-ray diffraction methods. The crystallographic R factor for the refined porcine aldehyde reductase model is 0.19 at 2.8 A resolution. There are two molecules in the asymmetric unit related by a local non-crystallographic twofold axis. The human aldehyde reductase model has been refined to an R factor of 0.21 at 2.48 A resolution. The amino-acid sequence of porcine aldehyde reductase revealed a remarkable homology with human aldehyde reductase. The coenzyme-binding site residues are conserved and adopt similar conformations in human and porcine aldehyde reductase apo-enzymes. The tertiary structures of aldhyde reductase and aldose reductase are similar and consist of a beta/alpha-barrel, with the coenzyme-binding site located at the carboxy-terminus end of the strands of the barrel. The crystal structure of porcine and human aldehyde reductase should allow in vitro mutagenesis to elucidate the mechanism of action for this enzyme and facilitate the effective design of specific inhibitors.

Journal Article↗
PubMed1994

[Surgery for acute arterial emboli to the arm].

During a 5-year period (1987-1992) we removed under local anesthesia 20 emboli and 2 thrombi from the arms of 18 patients aged 60-90 using a Fogarty catheter. 18 of the emboli (90%) were of cardiac origin. 1 patient had thrombosis of the axillary artery (2 episodes). The occlusions were brachial in 14 (70%), axillary in 5 (25%), and radial in 1. From 1.5 hours to 1 week elapsed between onset of symptoms and operation. 9 patients underwent arteriography. Surgery was successful in 19 (95%). Limb necrosis developed in a patient operated on 1 week after onset of symptoms. Necrosis progressed despite a palpable radial pulse. 33 patients (15%) died of causes unrelated to the operation: 2 had terminal cardiac disease and the third concomitant occlusion of the superior mesenteric artery. Early surgical exploration is advocated for acute arterial embolism of the upper extremity. Ischemic changes may be reversible even later than 24 hours from onset of symptoms. Routine arteriography is not needed before surgical exploration of these cases.

Aged↗
PubMed1994

Regulation of interleukin-8 expression in porcine alveolar macrophages by bacterial lipopolysaccharide.

Interleukin (IL)-8 is a macrophage-derived neutrophil chemotactic factor that plays an important role in the recruitment of neutrophils to inflammatory loci. Hence, expression of IL-8 by alveolar macrophages may be a significant factor in host defense in the lung and in the pathogenesis of pneumonia in swine. To initiate molecular studies of IL-8 regulation in pigs, we cloned IL-8 cDNA and examined the regulation of its mRNA in alveolar macrophages. The porcine IL-8 cDNA consists of 1491 base pairs including a coding region of 309 base pairs. The deduced amino acid sequence was 75 and 81% similar to human and rabbit IL-8, respectively. Resting macrophages contained low levels of IL-8 mRNA, which increased markedly after exposure to bacterial lipopolysaccharide (LPS). LPS induction of IL-8 was direct, not mediated through elevation of tumor necrosis factor or interleukin-1. The effect of LPS on IL-8 expression was dose dependent, and induction was observed at a concentration of 10 pg/ml. IL-8 mRNA expression was detectable within 0.5 h after stimulation with LPS, peaked at 3-6 h at about 30-fold higher levels than in resting cells, and was maintained for 24 h. Secreted IL-8, measured by neutrophil chemotaxis, was induced within 4 h by LPS, and accumulated in the media throughout the 24-h period. The mechanism of induction of IL-8 mRNA appeared to involve transcription and RNA processing. Nuclear run-on analysis showed that the IL-8 gene was actively transcribed in noninduced cells; upon stimulation with LPS, the rate of IL-8 transcription was increased about 4-fold. A single mature mRNA species was detected by primer extension analysis. The half-life of IL-8 mRNA transcripts in aveolar macrophages was approximately 2 h and did not change after LPS stimulation. The ability of LPS to induce IL-8 expression was suppressed by recombinant human IL-4 and dexamethasone in a concentration-dependent manner. These observations indicate that the expression of IL-8 is an early event in the sequelae to bacterial infection in the lung.

Amino Acid Sequence↗
PubMed1994

Interleukin-4 suppresses inflammatory cytokine gene transcription in porcine macrophages.

Inflammatory cytokines, including interleukin (IL)-1 alpha, IL-1 beta, IL-8, and tumor necrosis factor alpha (TNF-alpha) are produced by macrophages in response to a variety of pathogenic stimuli. We show here that the expression of inflammatory cytokines is suppressed by IL-4 at the transcriptional level. Interleukin-4, when added together with bacterial lipopolysaccharide (LPS), suppressed LPS-induced increases in mRNA levels of IL-1 alpha, IL-1 beta, IL-8, and TNF-alpha in alveolar macrophages. The level of suppression was dependent on dose and time of exposure and reached a maximum of 75-80% of uninduced values for IL-1 alpha, IL-8, and TNF. Interleukin-1 beta expression was completely inhibited by IL-4. The amount of secreted protein, as determined by TNF-alpha bioassay, was also suppressed by IL-4. Half-maximal suppression occurred at IL-4 concentrations between 0.02 and 0.1 ng/ml for all inflammatory cytokines. Nuclear run-on assays showed that IL-4 suppressed transcriptional activity of all inflammatory cytokines. Messenger RNA stability was not changed by IL-4. The data suggest that IL-4 plays an important transcriptional role in the regulation of alveolar macrophage inflammatory activities in respiratory disease and raise the possibility that IL-4 may function in vivo as a coordinator of inflammatory and immune responses.

Animals↗
PubMed1994

Decomposition of clozapine N-oxide in the qualitative and quantitative analysis of clozapine and its metabolites.

Pooled plasma from healthy volunteers was spiked with pure, synthetic clozapine or clozapine N-oxide and then made alkaline with either sodium hydroxide or sodium carbonate. The alkalized samples were allowed to stand at room temperature for various time intervals before extraction with organic solvent and then analyzed by high-performance liquid chromatography. It was found that clozapine N-oxide was reduced to clozapine in plasma made alkaline with sodium hydroxide, but such reduction was negligible in the plasma made alkaline with sodium carbonate. The amount of clozapine produced from clozapine N-oxide depended upon both the strength of the alkali added to the plasma and the duration of exposure of the plasma proteins plus clozapine N-oxide to the alkali. The reduction appears to take place through reducing equivalents generated by the action of strong alkali on plasma proteins. The thermal lability of clozapine N-oxide during gas chromatography-mass spectrometric analysis was also investigated. It was found that clozapine N-oxide was quantitatively decomposed to clozapine during gas chromatography-mass spectrometric analysis.

Chemistry Techniques, Analytical↗
PubMed1994

Differentiation of two classes of "A" system amino acid transporters.

Injection of mRNA from GF-14 cells (a mouse lymphocyte cell line) into Xenopus oocytes led to the expression of a transport activity characteristic of amino acid transport system A, i.e., Na+ dependent and recognizing aminoisobutyric acid and its methyl derivative (AIB and meAIB). Sucrose density gradient fractionation of GF-14 mRNA showed that the maximum level of expression of activity was associated with a 2.2-kb RNA fraction. Pretreatment of GF-14 cells with insulin caused a twofold increase in system A transport activity in the cells themselves and the mRNA from the insulin-treated cells induced a comparable increase in A system transport over control mRNA when expressed in oocytes. mRNA from cells treated with insulin in presence of actinomycin D did not show a response to insulin, suggesting that GF-14 cells synthesized new transporter in response to insulin. Similar experiments with mRNA from the Ehrlich ascites cells, showed that expression of system A type transport was associated chiefly with a 4.2-kb mRNA fraction. Although insulin treatment of Ehrlich cells also caused an increase in A system transport activity in the cells themselves (nearly twofold), the response to insulin was not blocked by actinomycin D or cycloheximide. Moreover, mRNA from insulin-treated and untreated Ehrlich cells gave the same level of expression in oocytes of A type amino acid transport. In contrast to GF-14 cells, Western blots of plasma membranes showed that insulin treatment of Ehrlich cells increased the amount therein of a 120- to 130-kDa peptide, previously associated with amino acid transport (Proc. Natl. Acad. Sci. USA 85, 7877, 1988), suggesting that in the Ehrlich cells, but not in GF-14, cells this polypeptide was translocated from intracellular sites in response to insulin. The data associate two distinctly different responses to insulin with A-type amino acid transporters and are consistent with the existence of more than a single A type amino acid transporter in mouse cells.

Amino Acids↗
PubMed1994

Comparison of personality traits of only and sibling school children in Beijing.

The number of only children in China has increased rapidly since the late 1970s, when the Chinese government advocated the "one-child-per-family" policy. This increase has caused much public concern about the healthy growth of the young generation. In this study the behavioral traits of Chinese, urban, only children were investigated. The participants were 444 children with siblings and 473 only children from Grades 1, 3, and 5 in Beijing primary schools. An inventory consisting of 27 items covering three behavioral traits was developed. The children were rated on the inventory by the principal teacher of the class. Factor analysis revealed three main factors: achievement motivation, interpersonal skills, and attitude toward manual labor. The only children exceeded the sibling children on achievement motivation. No differences were found between only children and sibling children on interpersonal skills and attitude toward manual labor. Gender differences were evident; girls consistently received higher ratings on achievement motivation and interpersonal skills. The possible underlying reasons for these differences are discussed.

Achievement↗
PubMed1994

Are elevated liver enzymes and bilirubin levels significant after laparoscopic cholecystectomy in the absence of bile duct injury?

OBJECTIVE: Increased aspartate aminotransferase (AST), alanine aminotransferase (ALT), and bilirubin levels were noted incidentally after a laparoscopic cholecystectomy. The percentage in which such elevation occurs and its clinical significance in the absence of bile duct injury were investigated. SUMMARY BACKGROUND DATA: Bile duct injury is the most feared complication of laparoscopic cholecystectomy. Some laboratory tests may be indicative of this complication, such as increases in liver enzyme (AST, ALT, and alkaline phosphatase [ALP]) and bilirubin. These parameters have not been investigated in patients who had laparoscopic cholecystectomy and in whom no damage to the bile duct was noted. METHODS: Sixty-seven patients with normal results of preoperative liver function test were entered into the study. Blood was collected 24 hours after laparoscopic cholecystectomy, and AST, ALT, ALP, and bilirubin levels were measured. RESULTS: A mean 1.8-fold increase in AST occurred in 73% of patients; 82% showed a 2.2-fold increase in ALT. A statistically nonsignificant increase was noted in 53% of patients (ALP remained within normal limits), and in 14% of patients bilirubin levels were increased (they were primarily of the unconjugated type). CONCLUSIONS: In many patients a significant increase in AST and ALT levels occurred after laparoscopic cholecystectomy, but they returned to normal values within 72 hours. The cause of this is unclear, and these elevations appear to have no clinical significance.

Adult↗