Biomedical subjects

F Chen

Publications and source records attributed to F Chen.

At least 235 records · Page 13Linked to original sources

PubMed2000

[Altered subcellular distribution of daunorubicin in the non-P-glycoprotein-mediated multidrug-resistant cell line HL-60/ADR].

OBJECTIVE: To investigate DNR subcellular distribution in the non-P-glycoprotein-mediated multidrug-resistant cell line HL-60/ADR and its relation to multidrug resistance. METHODS: DNR subcellular disposition was studied by confocal scanning laser microscopy, fluorescent methods, MTT and RT-PCR. The effects of verapamil, brefeldin A, chloroquine were also examined. RESULTS: In the drug-sensitive cell line HL-60 DNR fluorescence distributed evenly in the nucleus and cytoplasm, while in the resistant cell line DNR distributed in a punctate pattern in the cytoplasm and was reduced in the nucleus. Verapamil, brefeldin A, but not chloroquine could recover the intracellular distribution of DNR from punctate to even in the resistant cell line. CONCLUSION: Altered subcellular disposition of DNR in resistant cell line was involved in the mechanism of multidrug resistance.

Antibiotics, Antineoplastic↗

PubMed2000

[Regulation of telomerase activity in HL-60 and NB4 cells by arsenic trioxide].

OBJECTIVE: To investigate the regulation of telomerase activity in HL-60 and NB4 cells exposed to arsenic trioxide (As(2)O(3)). METHODS: Cell morphology, intracellular DNA distribution, bcl-2 expression and telomerase activity were evaluated in HL-60 and NB4 cells exposed to As(2)O(3) or ATRA. RESULTS: The differentiation induction of HL-60 and NB4 cells by low concentration of As(2)O(3) was weaker than that by ATRA, but downregulation effect of As(2)O(3) on telomerase activity was more strong and quick. Higher concentration of As(2)O(3) induced apoptosis in HL-60 or NB4 cells accompanied by more rapid down regulation of telomerase activity and bcl-2 expression. Neither As(2)O(3) nor ATRA showed direct inhibition effect on telomerase activity. CONCLUSION: The downregulation of telomerase activity by low concentration of As(2)O(3) in HL-60 and NB4 cells is the result of overlap of differentiation and apoptosis. There are similar mechanisms in the regulation of telomerase activity and apoptosis in different leukemia cells. Bcl-2 may play an important role in these mechanisms.

Apoptosis↗

PubMed2000

[Study on mixed stationary phase composed of 2,3,6-tributyl-beta-cyclodextrin and AgNO3 for capillary gas chromatography].

A new mixed gas chromatographic stationary phase composed of 2,3,6-tributyl-beta-cyclodextrin and AgNO3(in polyethylene glycol 400) was synthesized for the first time. It was coated on fused silica capillary column. Excellent selectivity was obtained for the separation of seven enantiomeric pairs and four groups of positional isomers of aromatic compounds. The recognition and separation mechanisms were discussed by measuring enthalpy, entropy, free energy and the difference in enthalpy and entropy of these compounds. Results were compared with those obtained on 2,3,6-tributyl-beta-cyclodextrin stationary phase. The results showed that the mixed stationary phase had positive synergistic effects.

Chromatography, Gas↗

PubMed2000

[Studies on gene knocking out of 2-keto aldose reductases from Erwinia sp. SCB125].

Based on the reported gene sequences, the segments containing 2-keto aldose reductase (2-KRA and B) genes were amplified by PCR from the plasmids and Erwinia sp. SCB125 each for gene expression and gene knocking out. Then cloning them into expression vector pBL and successfully expressing them with high enzyme activity in E. coli DH5 alpha. After their enzyme activities were proved, the work on gene knock out followed. Introducing the knock-out vector which distribute unstably during the cell division to the host strains Erwinia sp. SCB125. Screening firstly by the positive marker, one resistance which resulted from the expression of the resistance gene inserting inside the reductase genes and the negative marker, another resistance which outside the reductase genes in the vector. The strains selected out will be tested by further study. This work was the bases of blocking the pathway metabolism and constructing a recombinant strain that can produce 2-KLG directly from D-glucose by one-step fermentation.

Aldehyde Reductase↗

PubMed2000

[Supercritical extraction of Alpinia officinarum and GC-MS analysis of its components].

In order to find the effective components and new pharmaceutical effect of Alpinia officinarum, and further exploit it, we extracted its essential oils with the technique of Supercritical-CO2 fluid, then confirmed there were 27 components in Pot I And 111 components in Pot II by using the chemical component analysis and the computer information retrieval in the GC-MS system.

Alpinia↗

PubMed2000

[Clinical investigation of 0.3% levofloxacin eyedrops on the treatment of cases with acute bacterial conjunctivitis and bacterial keratitis].

PURPOSE: To investigate the effects of 0.3% levofloxacin eyedrops on the treatment of acute bacterial conjunctivitis and bacterial keratitis. METHODS: The patients were randomly divided into treated group and control group. They were given 0.3% levofloxacin and 0.3% ofloxacin eyedrops respectively. RESULTS: From April to December in 1999, 132 cases were enrolled in the study. It showed that there was no significant difference between two drops in the cure and effective rates, but the mean cure day of keratitis in levofloxacin group was significantly shorter than that in ofloxacin group. No serious side effects occurred. CONCLUSION: Levofloxacin eyedrops was effective and safe antibiotics in treating bacterial conjunctivitis and keratitis.

Adolescent↗

PubMed2000

[Partial laryngectomy plus radiotherapy versus partial laryngectomy alone for laryngeal carcinoma].

OBJECTIVE: To investigate the effect of radiotherapy in the combined treatment of laryngeal cancer with partial laryngectomy. METHODS: One hundred and seventy-one cases of laryngeal cancer were randomly separated into three groups of partial laryngectomy, partial laryngectomy with preoperative radiotherapy (dose 40-50Gy) and partial laryngectomy with postoperative radiotherapy(dose 51-70 Gy). The surgical procedures included cordectomy, vertical partial laryngectomy, horizontal partial laryngectomy and near total laryngectomy. RESULTS: The overall 5-year survival rate was 82% (123/150). The 5-year survival rates for surgery alone group (56 cases), surgery with preoperative radiotherapy group(65 cases) and surgery with postoperative radiotherapy group(50 cases) were 85.7%, 80.7% and 79.5% respectively. There was no significant difference among the groups(chi 2 = 0.703, P = 0.704). Analysis of survival rates revealed a significant difference among the different stages(chi 2 = 12.248, P = 0.007). All 171 cases except 3 that had near total laryngectomy achieved satisfactory phonation. CONCLUSION: Neither preoperative radiotherapy nor postoperative radiotherapy can improve the 5-year survival rate of laryngeal cancer with partial laryngectomy. All patients with partial laryngectomy got satisfactory phonation.

Adolescent↗

PubMed1999

Retina-specific nuclear receptor: A potential regulator of cellular retinaldehyde-binding protein expressed in retinal pigment epithelium and Müller glial cells.

In an effort to identify nuclear receptors important in retinal disease, we screened a retina cDNA library for nuclear receptors. Here we describe the identification of a retina-specific nuclear receptor (RNR) from both human and mouse. Human RNR is a splice variant of the recently published photoreceptor cell-specific nuclear receptor [Kobayashi, M., Takezawa, S., Hara, K., Yu, R. T., Umesono, Y., Agata, K., Taniwaki, M., Yasuda, K. & Umesono, K. (1999) Proc. Natl. Acad. Sci. USA 96, 4814-4819] whereas the mouse RNR is a mouse ortholog. Northern blot and reverse transcription-PCR analyses of human mRNA samples demonstrate that RNR is expressed exclusively in the retina, with transcripts of approximately 7.5 kb, approximately 3.0 kb, and approximately 2.3 kb by Northern blot analysis. In situ hybridization with multiple probes on both primate and mouse eye sections demonstrates that RNR is expressed in the retinal pigment epithelium and in Müller glial cells. By using the Gal4 chimeric receptor/reporter cotransfection system, the ligand binding domain of RNR was found to repress transcriptional activity in the absence of exogenous ligand. Gel mobility shift assays revealed that RNR can interact with the promoter of the cellular retinaldehyde binding protein gene in the presence of retinoic acid receptor (RAR) and/or retinoid X receptor (RXR). These data raise the possibility that RNR acts to regulate the visual cycle through its interaction with cellular retinaldehyde binding protein and therefore may be a target for retinal diseases such as retinitis pigmentosa and age-related macular degeneration.

Alternative Splicing↗

PubMed1999

Involvement of 5'-flanking kappaB-like sites within bcl-x gene in silica-induced Bcl-x expression.

The present study investigated the involvement of the transcription factor NF-kappaB in the expression of an anti-apoptotic gene, bcl-x, using a murine macrophage cell line and peritoneal macrophages from both wild type (p50(+/+)) and NF-kappaB p50 gene knockout (p50(-/-)) mice. Increased expression of Bcl-x protein was observed in native and silica-exposed p50(-/-) macrophages in which the NF-kappaB p65-containing complex was predominantly induced. Co-transfection experiment using a bcl-x promoter reporter construct and an expression vector for NF-kappaB p50 or p65 indicates that p65, but not p50, up-regulates the promoter activity of the bcl-x gene. DNA sequence analysis revealed that there are several kappaB-like sites within the 5'-flanking region of the bcl-x gene. Electrophoretic mobility shift assay suggested differences in binding of the NF-kappaB complexes to these putative NF-kappaB binding sites of the bcl-x gene.

Animals↗

PubMed1999

The human acid ceramidase gene (ASAH): structure, chromosomal location, mutation analysis, and expression.

Acid ceramidase (AC) is the lysosomal enzyme that degrades ceramide into sphingosine and fatty acid. A deficiency in human AC activity leads to the lysosomal storage disorder, Farber disease (FD). The human AC gene (HGMW-approved symbol ASAH) was cloned and characterized, revealing an organization similar to that of the murine AC gene. The human gene spans about 30 kb in length and contains 14 exons ranging in size from 46 to 1201 bp. The exon/intron junctions were determined and found to follow the GT-AG rule. The putative promoter region had a GC content over 60%, lacked a TATA box, and contained several sequences matching transcription factor binding sites, including nine SP-1 sites, one AP-1 site, and three CACC boxes. The promoter activity of a 475-bp fragment from within this region was demonstrated by chloramphenicol acyltransferase assays. Northern blotting revealed variable expression of the human AC RNA; i.e., expression of the major 2.4-kb transcript was high in heart and kidney, followed by lung and placenta, but low in pancreas, liver, brain, and skeletal muscle. Two minor AC transcripts of 1.7 and 1.2 kb also were detected in heart and skeletal muscle. The human AC gene was mapped to the chromosomal region 8p21.3-p22 by in situ hybridization and FISH analyses, syntenic with the mouse chromosomal location. Finally, three new missense mutations, E138V, R254G, and P362R, were identified in the human AC gene from FD patients. Mutant AC cDNAs containing these point mutations were constructed and examined using the FLAG-tagged expression system. Although the levels of protein expression for these mutant ACs were about equivalent to that of the controls, their enzymatic activity was markedly reduced, confirming their authenticity.

3T3 Cells↗

PubMed1999

Up-regulation of cortical AMPA receptor binding in the fawn-hooded rat following ethanol withdrawal.

The present study has employed quantitative receptor autoradiography to compare the binding of (S)-[3H]5-fluorowillardiine to (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors in the brains of alcohol-preferring Fawn-Hooded (FH) rats, alcohol non-preferring Wistar-Kyoto (WKY) rats, and FH rats following a 28-day period of 5% ethanol consumption with or without ethanol withdrawal. Significantly higher binding of [3H]5-fluorowillardiine was found in the cingulate cortex (+12%) and claustrum (+13%) in alcohol naïve FH rats compared to WKY rats. Chronic ethanol consumption decreased binding of (S)-[3H]5-fluorowillardiine in four cortical regions (frontal, parietal, occipital and temporal cortex), hippocampus and septohippocampal nucleus. In contrast, ethanol withdrawal induced a significant "rebound" increase in binding by +22% in frontal and parietal cortex, by +17% in cingulate cortex and +13% in claustrum, and by +14% in the septohippocampal nucleus compared to chronic ethanol-exposed FH rats. The findings suggest that AMPA receptors in frontal cortical regions are sensitive to ethanol and therefore may be implicated in the predisposition of alcohol preference in FH rats.

Alanine↗

PubMed1999

Induction of autoimmunity in a transgenic model of B cell receptor peripheral tolerance: changes in coreceptors and B cell receptor-induced tyrosine-phosphoproteins.

Abrogation of peripheral tolerance in transgenic mice that express a uniform B-cell receptor may create a powerful tool to examine the molecular mechanisms that underlie the autoimmune response in B cells. Here we report that processes that induce a systemic lupus erythematosus-like syndrome in normal mice, namely chronic graft vs host reaction, trigger systemic autoimmunity in a well-established transgenic mice model of B cell receptor peripheral tolerance. The induction of graft vs host reaction in mice that carry both a rearranged B cell Ag receptors specific for hen egg lysozyme and expressing chronically circulating hen egg lysozyme Ag resulted in induction of high and sustained levels of circulating anti-hen egg lysozyme autoantibodies and glomerulonephritis with proteinuria. This was associated with marked changes in expression of cell-surface proteins, such as CD23 and complement receptor 2. B cells from the graft vs host-induced mice could proliferate in vitro in response to self-Ag, and upon stimulation with anti-IgD demonstrated rapid phosphotyrosine phosphorylation of specific proteins, which could not be induced in the anergic double transgenic B cells. Conversely, loss of tolerance was not associated with a higher induction in the level of Syk kinase phosphorylation following stimulation with anti-IgD. Taken collectively, these data establish that 1) processes that induce a systemic lupus erythematosus-like syndrome in normal mice can abrogate peripheral tolerance in transgenic mice expressing self-tolerized B cells, and that 2) loss of tolerance in this model is associated with marked changes in surface expression of B cell coreceptors as well as with selective changes in IgD-induced signaling by discrete tyrosine-phosphoproteins, but not Syk kinase.

Animals↗

PubMed1999

RAG2 is regulated differentially in B and T cells by elements 5' of the promoter.

To study RAG2 gene regulation in vivo, we developed a blastocyst complementation method in which RAG2-deficient embryonic stem cells were transfected with genomic clones containing RAG2 and then assessed for their ability to generate lymphocytes. A RAG2 genomic clone that contained only the RAG2 promoter sequences rescued V(D)J recombination in RAG2-deficient pro-B cell lines, but did not rescue development of RAG2-deficient lymphocytes in vivo. However, inclusion of varying lengths of sequences 5' of the RAG2 promoter generated constructs capable of rescuing only in vivo B cell development, as well as other constructs that rescued both B and T cell development. In particular, the 2-kb 5' region starting just upstream of the RAG2 promoter, as well as the region from 2-7 kb 5', could independently drive B cell development, but not efficient T cell development. Deletion of the 2-kb 5' region from the murine germ line demonstrated that this region was not required for RAG expression sufficient to generate normal B or T cell numbers, implying redundancy among 5' elements. We conclude that RAG2 expression in vivo requires elements beyond the core promoter, that such elements contribute to differential regulation in the B vs. T lineages, and that sequences sufficient to direct B cell expression are located in the promoter-proximal 5' region.

Animals↗

PubMed1999

Freshly fractured crystalline silica induces activator protein-1 activation through ERKs and p38 MAPK.

The transcription factor activator protein-1 (AP-1) reportedly plays an important role in the induction of neoplastic transformation and multiple genes involved in cell proliferation, differentiation, and inflammation. To investigate the mechanisms of silica-induced carcinogenesis, AP-1-luciferase reporter transgenic mice were used as an in vivo model, whereas the JB6 mouse epidermal cell line and a rat lung epithelial cell line were employed as in vitro models to study the effects of silica at the molecular level. Freshly fractured silica caused an 8-fold increase in AP-1 activity in JB6 cells and a 2.5-fold increase in rat lung epithelial cells. The induction of AP-1 activity in cultured cell lines was time- and dose-dependent. Intratracheal administration of silica was also able to induce AP-1 transactivation in transgenic mice. AP-1 activation was first observed at 2 days after silica administration and reached its maximum at 3 days post-exposure of the mice to silica. The signal transduction pathways for AP-1 activation were also investigated using these cell lines. The results demonstrate that freshly fractured silica stimulates mitogen-activated protein kinase (MAPK) family members, as determined by the phosphorylation of p38 MAPK and extracellular signal-regulated protein kinases (ERKs). Inhibition of ERKs with PD98059 or of p38 with SB203580 significantly inhibited silica-induced AP-1 activation. These findings demonstrate for the first time that freshly fractured silica induces AP-1 activation, which may be mediated through p38 MAPK and ERK pathways. Unraveling the complex mechanisms associated with these events may provide insights into the initiation and progression of silica-induced carcinogenesis.

Animals↗

PubMed1999

Identification of 2,3-butanedione monoxime hydrogenation products by gas chromatography-mass spectrometry in an ion trap mass spectrometer.

The reduced products of 2,3-butanedinone monoxime by reaction with hydrogen in the presence of homogeneous catalysts were identified by gas chromatography coupled to an ion trap mass spectrometer operating either in the electron impact or chemical ionization mode. The major hydrogenation products were found to be several heterocyclic nitrogen-containing compounds: tetramethylpyrazine, 2,4-dimethyl-3-ethylpyrrole, 3,4,5-trimethylpyrazole, 2,5-dimethyl-1-propylpyrrole, 3-acetyl-2,4-dimethylpyrrole, 3,5-dimethyl-4-allypyrazole and tetramethylpyrazine N-monoxide.

Diacetyl↗

PubMed1999

Adaptive numerical morphological filter for identifying chromatographic signals.

A morphological algorithm has been proposed to filter away impulsive noises confounded in the chromatographic signal. Compared with the conventional median filtering method, the results showed that the proposed method has the advantages of a better filtering effect and less distortion. In particular, the morphological filter with adaptive scale gives very good results.

Algorithms↗

PubMed1999

Multiple glucocorticoid receptor transcripts in membrane glucocorticoid receptor-enriched S-49 mouse lymphoma cells.

A cDNA library from plasma membrane glucocorticoid receptor-enriched (mGR(++)) S- 49 mouse T lymphoma cells was screened with full-length rat intracellular GR (iGR) cDNA, BUGR-2 antibody, and PCR amplimers to portions of the mouse GR cDNA. One or two single-base substitutions resulting in amino acid changes (which do not incapacitate the receptor) were found in all but one clone: Val437 --> Gly (located in the first zinc finger), and Glu546 --> Gly (in the steroid-binding domain). Two previously unidentified exon 1 variants (1D and 1E), and two of three previously reported variants (1A, 1B) were found to be spliced onto the common exon 2. Exon 1D- and 1E-containing transcripts were confirmed by direct sequencing of amplimers from reverse transcriptase-coupled PCR. RNase protection studies revealed that one of these transcripts was expressed in mGR(++) cells only, but not in two mGR-less (mGR(--) S-49, and AtT-20 mouse pituitary) cell lines. These studies suggest that at least four promoters may be responsible for the control of GR (iGR and mGR) types in mouse lymphoma cells.

5' Untranslated Regions↗

PubMed1999

Association of the glucocorticoid receptor alternatively-spliced transcript 1A with the presence of the high molecular weight membrane glucocorticoid receptor in mouse lymphoma cells.

Using the combination of a cDNA library prepared from membrane glucocorticoid (mGR)-enriched S-49 cells and a mouse leukocyte genomic library, we have cloned a 7.3 kb full-length glucocorticoid receptor 1A cDNA. Primer extension, 5'RACE, and long distance PCR identified the transcription start site as being located at 1026 bp from the ATG codon. The first 1,013 nucleotides (nts) of the full length sequence constitute 5' UTR sequence (exon 1), the next 2349 bp, the coding region, and the last 3,907 bp, the 3'UTR. The entire 5'UTR sequence is unique to transcript 1A. The 3'UTR sequence is approximately 88.5 % conserved with the rat 3'UTR. Western blot analysis compared the molecular weight of in vitro translation products from the cloned 1A cDNA with partially purified cellular mGR. Both preparations contained the novel 150 KD and the 94 KD classical GR peptides, suggesting that transcript 1A encodes both receptor forms. Transfection of mGR-less and glucocorticoid lysis-resistant AtT-20 and HL-60 cells with full-length GR 1A cDNA imparted both mGR expression and glucocorticoid lysis-sensitivity to these cells.

3' Untranslated Regions↗