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Y Dong

Publications and source records attributed to Y Dong.

At least 271 records · Page 15Linked to original sources

The glucocorticoid receptor and a putative repressor protein coordinately modulate glucocorticoid responsiveness of the mouse mammary tumor virus promoter in the rat hepatoma cell line M1.19.

Signal transduction by glucocorticoid hormones is mediated by the intracellular glucocorticoid receptor protein. The mechanisms determining cell type- or tissue-specific differences in hormone responsiveness remain, however, unclear. To address this issue we have used two different rat hepatoma cell lines, 762 and 6.10.2, respectively, in which mouse mammary tumor virus has been stably integrated. Nuclear extracts from both of these cell lines contained a factor that bound to a sequence motif extending from -163 to -147 in the mouse mammary tumor virus promoter and that appeared to repress hormonal induction of viral mRNA expression. Transient transfection experiments indicated that the cellular levels of this putative repressor did not affect basal promoter activity; this factor appeared rather to determine cellular sensitivity to glucocorticoids. Moreover, in these experiments the relative levels of the glucocorticoid receptor appeared to be the main determinant of maximum inducibility of virus expression by hormone. Taken together, these data indicate that the differential expression patterns of receptor versus the putative repressor protein may determine the level of hormonal responsiveness of target genes in glucocorticoid-sensitive tissues.

Animals↗

A new macrophage differentiation antigen which is a member of the scavenger receptor superfamily.

The antibodies Ki-M8, Ber-Mac3, GHI/61 and SM4 define a human macrophage-associated antigen with a relative molecular mass of 130,000 which we designate M130. The protein was purified by immunoaffinity chromatography and an N-terminal and three internal amino acid sequences were obtained. A cDNA fragment was initially obtained by polymerase chain reaction (PCR) using reverse-translated primers. Several variant cDNA clones, derived from alternative spliced messages, were obtained from a lipopolysaccharide-stimulated human monocyte library and were sequenced. The relative abundance of these variants was evaluated by a series of overlapping PCR reactions. The size of the most representative cDNA is 3.7 kb and closely agrees with the mRNA size of 3.8 kb determined by Northern blot analysis. The membrane protein encoded contains a leader peptide of 40 residues, a putative extracellular domain of 1003 residues, followed by a hydrophobic segment of 24 residues and a cytoplasmic domain of 49 residues. The extracellular domain was found to contain nine repeating elements, of about 110 residues, which are similar to those of the scavenger receptor superfamily.

Amino Acid Sequence↗

Cereal grain digestion by selected strains of ruminal fungi.

The ruminal fungi Orpinomyces joyonii strain 19-2, Neocallimastix patriciarum strain 27, and Piromyces communis strain 22 were examined for their ability to digest cereal starch. All strains digested corn starch more readily than barley or wheat starch. Orpinomyces joyonii 19-2 exhibited the greatest propensity to digest starch in wheat and barley, whereas the digestion of these starches by N. patriciarum 27 and P. communis 22 was limited. Media ammonia concentrations were lower when fungal growth was evident, suggesting that all strains assimilate ammonia. Fungi formed extensive rhizoidal systems on the endosperm of corn, but O. joyonii 19-2 was the only strain to form such systems on the endosperm of wheat and barley. All strains penetrated the protein matrix of corn but did not penetrate starch granules. Starch granules from all three cereals were pitted, evidence of extensive digestion by extracellular amylases produced by O. joyonii 19-2. Similar pitting was observed on the surface of corn starch granules digested by N. patriciarum 27 and P. communis 22, but not on wheat and barley starch granules. The ability of ruminal fungi to digest cereal grains depends on both the strain of fungus and the type of grain. The extent to which fungi digest cereal grain in the rumen remains to be determined.

Ammonia↗

Comparison of amylolytic and proteolytic activities of ruminal fungi grown on cereal grains.

Strains of the ruminal fungi Neocallimastix patriciarum, Orpinomyces joyonii, and Piromyces communis were grown on cellobiose and on cereal grains and then examined for proteolytic and amylolytic activities. On cellobiose all three fungi displayed similar activities, with the exception of little amylolytic activity in the cell-associated fraction of N. patriciarum. Growth on the cereal grains barley, corn, and wheat showed differences in proteolytic and amylolytic activities amongst the ruminal fungi and between the cereal grains. The data suggest that while these fungi are capable of fermenting the cereal grains the mode of enzymatic attack varies both with the particular fungus and the type of cereal grain.

Amylases↗

Detection of anti-SSB antibodies in patients with rheumatic diseases.

Anti-SSB antibodies were measured by ELISA in patients with various kinds of connective tissue diseases using SSB antigen purified from fresh rabbit thymus. The SSB antigen reacted with anti-SSB standard serum, and the positive rates in SS, SLE, RA, PBS and MCTD were 55.1%, 48.3%, 32.8%, 30.8% and 26.3%, respectively. The titers of anti-SSB antibodies were higher in SS and SLE patients than in other connective tissue disease patients. However, 10% of normal individuals were found to have anti-SSB antibodies with low titers. The anti-SSB antibodies detected were mainly of IgG isotype. Preliminary analysis of clinical data showed no relationship between anti-SSB and systemic involvement in SS.

Antibodies, Antinuclear↗

[The purification of SS-B antigen and detection of anti-SS-B antibodies].

SS-B antigen was purified from fresh rabbit thymus by ammonium sulfate precipitation and column chromatography with Sephadex G100 and phosphocellulose. The M. W. of SS-B is ranged at 41,000 to 48,000. It does not contain the other extractable antigens, like Sm, RNP, PM-ScL, Scl-70, Jo-1, and PCNA. The purified SS-B antigen only reacts with the CDC standard serum of anti-SS-B antigen only reacts with the CDC standard serum of anti-SS-B antibody by ELISA. The positive rate of the antibodies being 55.1%, 48.3%, 32.8%, 30.8% and 26.3% in SS, SLE, RA, PSS and MCTD respectively. The titers of anti-SS-B antibodies were higher in SS and SLE patients than other connective tissue disease patients. It was found that all of the anti-SS-B antibodies detected were mainly of IgG isotype. Preliminary analysis of clinical date shows that there is no relationship between anti-SS-B antibody and systemic involvement in SS.

Arthritis, Rheumatoid↗

[Sensitivity in vitro of Plasmodium falciparum to chloroquine, pyronaridine, artesunate and piperaquine in south Yunnan].

The sensitivity of P. falciparum to chloroquine, pyronaridine, artesunate and piperaquine (CQ, PD, AT, PQ) was assayed using in vitro microtechnique in south Yunnan in 1990. The resistance rates were 98.7% (75/76), 27.6% (16/58), 13.8% (9/65) and 97.7% (43/44) respectively, and ID50 were 125.0, 19.0, 4.7 and 243.3 nmol/L, respectively. The resistance rate against CQ showed no change as compared to the rates against CQ 5 and 9 years ago; but the ID50 was lower. CQ-resistant P. falciparum showed a marked cross-resistance to PQ, but not to PD and AT. AT-resistant P. falciparum exhibited cross-resistance to the above-mentioned three drugs. PD-resistant P. falciparum showed no cross resistance to AT, but showed cross resistance to CQ and PQ. In comparison with chloroquine-coated plates, the plates coated with pyronaridine, artesunate or piperaquine gave similar results as the former, which were shown by the rise in schizont inhibition rates along with the rise in drug concentration. It indicates that pyronaridine-, artesunate-, and piperaquine-coated plates can be used in the assay of sensitivity of P. falciparum to the three drugs.

Animals↗

[The manifestations of the nervous system in primary Sjogren syndrome].

We reported 9 cases of primary Sjogren syndrome (SS) who were complicated with nervous system involvement. All were women. Age between 24 to 58 years old. Their clinical symptoms of the nervous system varied widely, 7 of them manifested with the involvement of central nervous system, the main features were in case 1 bouts of seizures, case 2 multi-level damage of the brain and spinal cord which was similar to multiple sclerosis, case 3 recurrent hemiplegia caused by cerebral thrombosis then fatal vascular hemorrhage, case 4 sudden hemiplegia, case 5 persistent psychological disturbance, case 6 cranial neuropathy of V and VII, case 9 multi-focal symptoms of ataxia, myelopathy and transient blindness. 5 of the 9, case 3, 6-9 all appeared with the symptoms of peripheral neuropathy, case 7 also complicated with carpal tunnel syndrome. 4 of the 9 cases also complicated with renal tubular acidosis and/or chronic active hepatitis and/or fibrosing alveolitis and/or thrombocytopenic purpura and/or myositis. No differences of the positivity of autoantibodies was observed between those with or without nervous system involvement.

Adult↗

Identification and characterization of a cis-acting element that interferes with glucocorticoid-inducible activation of the mouse mammary tumor virus promoter.

The rat hepatoma cell line M1.19 is stably infected by the mouse mammary tumor virus (MMTV), and the expression of the virus is induced by glucocorticoid treatment. However, in the 6.10.2 variant of M1.19, an increase in MMTV transcription is hardly detectable upon exposure to hormone. The mechanism of hormone-unresponsiveness in these cells has been unclear. In this study, we show that nuclear extract from 6.10.2 cells contains a specific DNA-binding activity that recognizes a sequence motif extending from positions -163 to -147 on the MMTV promoter. An oligonucleotide probe spanning this region binds a nuclear factor distinct from the glucocorticoid receptor. In vivo competition experiments, where increased amounts of a plasmid containing this element were transfected into 6.10.2 cells, showed a dose-dependent increase in hormonal inducibility of MMTV expression. Together, these results indicate that this sequence motif negatively modulates glucocorticoid-inducible activation of the MMTV promoter. Moreover, we have characterized a nuclear factor that preferentially binds to the coding strand of this element.

Animals↗

Regulation of glucocorticoid receptor expression.

A limiting factor determining the sensitivity of a cell to glucocorticoid hormones is the intracellular concentration of the glucocorticoid receptor (GR) protein. By regulating the expression of GR the cell is able to adapt to both changes in its hormone environment and to the varying requirements for biological response. Studies on the regulation of GR expression have shown this to be a complex process which involves both transcriptional and posttranscriptional mechanisms. Although GR is more or less constitutively expressed in most tissues its concentration varies under different physiological conditions. GR expression is regulated by a number of different agents including factors which act through a second messenger pathway. This allows the cell to control glucocorticoid regulated gene expression through a complex but integrated hormonal network. Here we summarize our studies on GR regulation with emphasis on: i), GR autoregulation; ii), the effect of cAMP on GR expression, and iii), GR expression during fetal rat lung development.

Animals↗

Inhibition of mitochondrial respiratory function by an organic solvent extractable component from an extract of burn eschar.

Saline extracts of burn eschar (CEBE) and normal skin (CENS) caused inhibition to mitochondrial respiration and inner membrane function. Ethyl acetate extracts from CEBE (D1) and CENS (D'1) caused depression of the Respiratory Control Ratio, (RCR), an inhibition of respiration rate in state 3 and stimulation to state 4 respiration. Excellent linear correlations exist between the degree of inhibition to state 3, rate of stimulation to state 4 respiration and the logarithm of doses of D1 and D'1. The effective dose ranges (0.75-0.25 mg/ml for D1 and 4-1 mg/ml for D'1) differ by one order of magnitude. The activity of NADH dehydrogenase and succinate dehydrogenase of mitochondria after incubation with the highest toxic dose of D1 or D'1 remained normal. Dinitrophenol (DNP)-stimulated respiration was moderately inhibited by D1 and D'1. No change of oligomycin-sensitive ATPase activity was demonstrated. Exogenous malondialdehyde (MDA) did not show any inhibitory effect. Preliminary studies show that D1 contains a family of free fatty acids (FFA). Incubation of normal mitochondria with D1 increased the content of saturated FFA and a decrease of unsaturated FFA. The role of other peroxidative products is under investigation.

Adenosine Triphosphatases↗

The mechanism for glucocorticoid-resistance in a rat hepatoma cell variant that contains functional glucocorticoid receptor.

The mechanism of glucocorticoid resistance has been studied in a rat hepatoma cell variant (6.10.2), which contains low levels of glucocorticoid receptor (GR). These cells seem to have lost all the glucocorticoid-induced transcriptional responses as measured by the lack of induction of expression of stably integrated mouse mammary tumor virus (MMTV) and the endogenous gene tyrosine aminotransferase (TAT), as well as the transcriptional suppression of GR gene expression. Physico-chemical characterization of the GR in the glucocorticoid resistant 6.10.2 cells revealed that the receptor is indistinguishable from the wild-type receptor with regard to size, hormone- and DNA-binding. The levels of the receptor mRNA and the total immunoreactive protein found in 6.10.2 cells were about 20% of those found in wild-type cells. Further analysis of 6.10.2 cells demonstrated that the receptor was indeed biologically functional. Treatment of 6.10.2 cells with 8-bromo-cAMP, which induced the endogenous GR level two-fold, restored responsiveness to glucocorticoids. Secondly, pretreatment of the cells with cycloheximide also led to reacquisition of cellular responsiveness to glucocorticoids. We propose that there exists a "threshold" level of GR, which is required for responsiveness and that under normal culture conditions, the level of GR in 6.10.2 cells is below this threshold. Glucocorticoid responsiveness can be restored by raising the GR level above the threshold with 8-bromo-cAMP or, alternatively, by removing the threshold barrier (repressor protein) with cycloheximide. Finally, the existence of such a repressor protein for MMTV induction was shown by in vivo titration with an isolated negative cis-element from the MMTV promoter.

8-Bromo Cyclic Adenosine Monophosphate↗

Prostaglandin E1 activates a chloride current in Jurkat T lymphocytes via cAMP-dependent protein kinase.

Patch-clamp studies have identified a cAMP-dependent Cl- conductance in lymphocytes that is defectively regulated in cystic fibrosis. In this study we used 125I efflux and whole-cell patch-clamp studies to investigate whether prostaglandin E1 (PGE1), an agonist that generates intracellular cAMP in Jurkat T lymphocytes, activates a Cl- conductance. Stimulation of T cells by externally applied PGE1 stimulated 125I efflux and activated a slowly developing membrane current. When external and internal Cl- were about equal, the current reversed at about zero mV, but when external Cl- was lowered from 157 to 7 mM the reversal potential shifted 75 mV in the positive direction, demonstrating that the current carrier was Cl-. In addition, the current was blocked by 10 microM 5-nitro-2(3-phenylpropylamino) benzoic acid (NPPB), a potent Cl- channel blocker. A membrane-permeable cAMP analog mimicked the effect of PGE1, whereas intracellular application of a cAMP antagonist Rp-cAMP blocked the effect of PGE1. Addition of purified catalytic subunit of cAMP-dependent protein kinase (PKA) plus ATP to the recording pipette also activated a similar current, whereas internally applied Walsh inhibitor, the synthetic peptide inhibitor of PKA, blocked the PGE1 effect. These results suggest that PGE1, acting through PKA, activates a Cl- current in Jurkat T cells.

Alprostadil↗