[Wound healing and extracellular matrix].
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Animals↗
Biomedical subjects
Z Xia
Publications and source records attributed to Z Xia.
[The check and trouble shoot of mA trouble of shimadzu X-ray generator HD150B-10].
X-ray machine mA circuits are generally composed by mA Pre-set circuits and mA detect circuits. To effectively control the tube current, there are many safety circuits and auxiliary circuits in mA control circuits. The trouble of mA circuits of HD150B-10 displays "mA Trouble" promptly on the plasma display; however, it is difficult to discover the actual trouble in the circuits because the mA control circuits are interconnected. In this paper, the question we discuss is how to check and shoot quickly and correctly the trouble of mA circuits, after the prompt appearance of "mA Trouble" on the plasma display.
Equipment Design↗
[Comparative study of two fusion retinoic acid receptor alpha in acute promyelocytic leukemia].
OBJECTIVE: To make a comparison of biological properties between PML-RARalpha and PLZF-RARalpha fusion proteins. METHODS: Receptor radioligand binding assay, receptor DNA binding assay and immunofluorescence methods were used. RESULTS AND CONCLUSION: PML-RARalpha and PLZF-RARalpha had similar ligand-binding affinities. Both could bind in vitro to retinoic acid response elements (RAREs) forming homodimers and to retinoic acid X receptor. However ,they differed in the relative binding affinities to different RAREs,the behavior of forming complex with RXR and the subcellular localization. More importantly, PML-RARalpha and PLZF-RARalpha could block different regulatory pathways mediated by PML or PLZF, through heterocomplex formation with wild-type PML or PLZF. The differences between PML-RARalpha and PLZF-RARalpha may in part explain the apparent resistance of APL with t(11;17) to differentiating effect of all-trans retinoic acid (ATRA).
DNA↗
[The preliminary study on prevention and treatment of neonatal jaundice].
OBJECTIVE: The purification and identification of heme oxygenase isoforms (HO-1) and HO-2 from human liver were described and Sn-protoporphyrin (SnPP) was used to inhibit HO-1 activity in order to provide a new method for prevention and treatment of neonatal jaundice. METHODS: Human hepatic microsomal fractions were purified by DEAE-Sephadex A-25 and hydroxylapatite chromatography. The enzyme activities of the two isoforms were detected with and without Snpp. RESULTS: HO-1 was the predominant form with a ratio of 2.4:1 (HO-1:HO-2). The apparent molecular weight of HO-1 and HO-2 on SDS-PAGE was 30,000 and 36,000 under reducing conditions, respectively. The study also showed that HO-1 was the traditional inducible heme oxygenase and HO-2 was a non-inducible heme oxygenase. CONCLUSION: The efficient inhibition effect of SnPP on HO-1 is suggested.
Enzyme Inhibitors↗
Calcium influx via the NMDA receptor induces immediate early gene transcription by a MAP kinase/ERK-dependent mechanism.
The regulation of gene expression by neurotransmitters is likely to play a key role in neuroplasticity both during development and in the adult animal. Therefore, it is important to determine the mechanisms of neuronal gene regulation to understand fully the mechanisms of learning, memory, and other long-term adaptive changes in neurons. The neurotransmitter glutamate stimulates rapid and transient induction of many genes, including the c-fos proto-oncogene. The c-fos promoter contains several critical regulatory elements, including the serum response element (SRE), that mediate glutamate-induced transcription in neurons; however, the mechanism by which the SRE functions in neurons has not been defined. In this study, we sought to identify transcription factors that mediate glutamate induction of transcription through the SRE in cortical neurons and to elucidate the mechanism(s) of transcriptional activation by these factors. To facilitate this analysis, we developed an improved calcium phosphate coprecipitation procedure to transiently introduce DNA into primary neurons, both efficiently and consistently. Using this protocol, we demonstrate that the transcription factors serum response factor (SRF) and Elk-1 can mediate glutamate induction of transcription through the SRE in cortical neurons. There are at least two distinct pathways by which glutamate signals through the SRE: an SRF-dependent pathway that can operate in the absence of Elk and an Elk-dependent pathway. Activation of the Elk-dependent pathway of transcription seems to require phosphorylation of Elk-1 by extracellular signal-regulated kinases (ERKs), providing evidence for a physiological function of ERKs in glutamate signaling in neurons. Taken together, these findings suggest that SRF, Elk, and ERKs may have important roles in neuroplasticity.
Animals↗
Determination of the gene sequence and the three-dimensional structure at 2.4 angstroms resolution of methanol dehydrogenase from Methylophilus W3A1.
The DNA sequences for the genes encoding the heavy and light subunits of methanol dehydrogenase from Methylophilus methylotrophus W3A1 have been determined. The deduced amino acid sequence has enabled the structure of the enzyme to be refined at 2.4 angstrom resolution against X-ray data collected on a Hamlin area detector. The structure was refined using the programs PROFFT and X-PLOR with several model building step interspersed. The final model contains two heavy chains (571 amino acids), two light chains (69 amino acids), two molecules of pyrroloquinoline quinone, two Ca2+ and 521 solvent molecules. Each half molecule contains four disulfide linkages and four cis peptides. One of the disulfides is formed from two adjacent cysteine residues linked by a trans peptide which creates a novel eight-membered ring. The heavy subunit is an 8-fold beta-propeller, each "blade" of which is a four-stranded antiparallel twisted beta-sheet. The light chain is an elongated subunit stretching across the surface of the heavy subunit, with residues 1 to 32 containing four beta-turns and residues 33 to 62 forming a helix; however, it neither interacts with the active site, nor the other HL dimer and its functional role is obscure. Around the 8-fold beta-propeller there is a repeating pattern of tryptophan residues located in the outer strand of seven of the eight beta-leaflets, each packed between adjacent leaflets. Each of these tryptophan residues is centered in the beta-strand and participates in the main chain hydrogen bonding of the sheet. Five of the seven tryptophan residues have closely similar interactions with the adjacent beta-leaflet including stacking of the tryptophan indole rings against a peptide plane and formation of a hydrogen bond from NE1 of the indole ring to a main-chain carbonyl. This repeating pattern is conserved over a number of MEDH sequences. The PQQ is located on the pseudo 8-fold rotation axis of the heavy subunit, in a funnel-shaped internal cavity, sandwiched between the indole ring of Trp237 and the two sulfur atoms of the Cys103-Cys104 vicinal disulfide. A hexacoordinate Ca2+ is bound in the active site by one nitrogen and five oxygen ligands, three from the PQQ and the others from two protein side-chains. In the active site an isolated solvent molecule is bound to the O5 of PQQ and to a nearby aspartate side-chain; its position may be the binding site for methanol. The aspartate might than serve as a general base for proton abstraction from the substrate hydroxyl. The C5 atom of PQQ could be activated by electrophilic catalysis by a nearby argenine side-chain or by the calcium ion bound to PQQ.
Alcohol Oxidoreductases↗
Post-transcriptional regulation of chymase expression in mast cells. A cytokine-dependent mechanism for controlling the expression of granule neutral proteases of hematopoietic cells.
Although all mouse mast cells are derived from a common progenitor, these effector cells exhibit tissue-specific differences in their expression of the chymase family of serine proteases whose genes reside on chromosome 14. Immature bone marrow-derived mast cells (mBMMC), developed in vitro with interleukin (IL) 3-enriched medium, were cultured in the presence or absence of IL-10 to determine at the molecular level how the expression of the individual chymases is differentially regulated. As assessed by RNA blot analysis, mBMMC contain high steady-state levels of the transcript that encodes mouse mast cell protease (mMCP) 5, but not the homologous chymase transcripts that encode mMCP-1, mMCP-2, or mMCP-4. Nevertheless, nuclear run-on analysis revealed that these cells transcribe all four mast cell chymase genes. IL-10 elicited high steady-state levels of the mMCP-2 transcript, and pulse-chase experiments revealed that the half-life of the mMCP-2 transcript in mBMMC maintained in the presence of IL-10 is approximately 4-fold longer than that in replicate cells subsequently cultured in medium without IL-10. Reverse transcription-polymerase chain reaction/nucleotide sequence analysis demonstrated that mBMMC cultured in the absence or presence of IL-10 correctly process mMCP-2 pre-mRNA. Experiments with cycloheximide and actinomycin D indicated that IL-10 induces expression of a trans-acting factor(s) that stabilizes the mMCP-2 transcript or facilitates its processing. The discovery that the expression of certain chymases in mBMMC is regulated primarily at the post-transcriptional level provides a basis for understanding the mechanism by which specific cytokines dictate expression of the chromosome 14 family of serine proteases in cells that participate in inflammatory processes.
Animals↗
Natural disruption of the mouse mast cell protease 7 gene in the C57BL/6 mouse.
The C57BL/6 mouse differs from the BALB/c mouse in that its ear and skin mast cells and its progenitor bone marrow-derived mast cells (mBMMCs) do not express mouse mast cell protease (mMCP) 7. We now report that, as detected by nuclear run-on analysis, the mMCP-7 gene is transcribed in C57BL/6 mBMMCs at a rate comparable to that in BALB/c mBMMCs. Reverse transcriptase-polymerase chain reaction analysis and sequencing of the product revealed that the ears of C57BL/6 mice contain small amounts of a mMCP-7 transcript that possesses a 98-base pair deletion. The deletion begins at a normally quiescent cryptic splice site (G416TGAG), 98 base pairs upstream of the normal exon 2/intron 2 splice site (G514TGAG), and introduces a premature stop codon in the alternatively spliced transcript. Thus, even if translated, the mature protein would consist of only 18 amino acids as compared to 245 amino acids in normal mMCP-7. Sequence analysis of the mMCP-7 gene in the C57BL/6 mouse revealed that the cryptic splice site is activated due to a G514-->A point mutation at the first nucleotide of the normal exon 2/intron 2 splice site. This is the first report of a mutation of a gene that encodes a mast cell secretory granule constituent that leads to its loss of expression. Moreover, the mMCP-7 gene is the first found in any species that sequentially has undergone a splice site mutation to cause retention of an intron and then a second splice site mutation to cause activation of a cryptic splice site.
Amino Acid Sequence↗
Dysregulation of bcl-2, c-myc, and Fas expression during tricyclic antidepressant-induced apoptosis in human peripheral lymphocytes.
Tricyclic antidepressants (TCAs) have been shown to induce apoptosis in human lymphocytes. In the present report, we investigated in parallel the regulation of the three oncogenes bcl-2, c-myc, and Fas. A reduction in c-myc and bcl-2 levels of 35-40% and 22-27%, respectively, was observed. On the other hand, Fas expression on the outer surface of the plasma membrane was increased up to 31%. In conclusion, bcl-2, c-myc, and Fas are undergoing dysregulation due to TCA-induced apoptosis.
Antidepressive Agents, Tricyclic↗
Tricyclic antidepressants inhibit IL-6, IL-1 beta and TNF-alpha release in human blood monocytes and IL-2 and interferon-gamma in T cells.
Tricyclic antidepressants (TCAs) are widely used in treating depressive disorders. It has been demonstrated that, for instance, IL-1 beta and IL-6 inhibit the HPA axis, which plays a role in the development of depressions. Therefore. we were interested in investigating how TCAs influence cytokine release by T lymphocytes and monocytes respectively. Cells were incubated with either 5 microM clomipramine, 15 microM imipramine or 20 microM citalopram. IL-2 release was suppressed to 60% of the control values by clomipramine and imipramine (p = 0.001; p = 0.000), but citalopram was found to cause a much weaker inhibition (only 18%) (p = 0.16). INF-gamma release was affected to a lower degree than IL-2 release, and imipramine (34%) (p = 0.054) was more potent than clomipramine (24%) (p = 0.16) and citalopram (12%) (p = 0.059) in this case. Monocytes incubated with TCA for 4 h exhibited only limited inhibition of IL-1 beta and IL-6 release, i.e., 6-25% for all three compounds. The corresponding value for TNF-alpha release was 20-45% inhibition, with citalopram being the weakest inhibitor. After 10 h of monocytes to LPS exposure, all three compounds exerted a strong inhibition of IL-1 beta and TNF-alpha release, i.e., 60-70% with p-values below 0.012 for all of them. However the inhibition of IL-6 release was less than 35%. Citalopram was equality as potent as imipramine and clomipramine in inhibiting IL-6 release after long-term exposure of monocytes to LPS. All three TCAs elevated intracellular cAMP concentrations significantly in T lymphocytes and monocytes (p < 0.001).
Antidepressive Agents, Tricyclic↗
The risk of pregnancy after tubal sterilization: findings from the U.S. Collaborative Review of Sterilization.
OBJECTIVE: Our purpose was to determine the risk of pregnancy after tubal sterilization for common methods of tubal occlusion. STUDY DESIGN: A multicenter, prospective cohort study was conducted in U.S. medical centers. A total of 10,685 women who underwent tubal sterilization was followed up for 8 to 14 years. The risk of pregnancy was assessed by cumulative life-table probabilities and proportional hazards models. RESULTS: A total of 143 sterilization failures was identified. Cumulative 10-year probabilities of pregnancy were highest after clip sterilization (36.5/1000 procedures) and lowest after unipolar coagulation (7.5/1000) and postpartum partial salpingectomy (7.5/1000). The cumulative risk of pregnancy was highest among women sterilized at a young age with bipolar coagulation (54.3/1000) and clip application (52.1/1000). CONCLUSIONS: Although tubal sterilization is highly effective, the risk of sterilization failure is higher than generally reported. The risk persists for years after the procedure and varies by method of tubal occlusion and age.
Age Factors↗
Optimally functional fluorescein isothiocyanate-labelled fibrinogen for quantitative studies of binding to activated platelets and platelet aggregation.
Dynamic and quantitative studies of the binding of fibrinogen (Fg) to its receptor, GPIIb-IIIa, on activated platelets, leading to platelet aggregation, are best studied with fluorescently-labelled Fg by flow cytometry. Due to conflicting reports on the functionality of FITC-labelled Fg, we have developed a reproducible and 'mild' labelling of fibrinogen with FITC-celite at pH 7.4-8.5 for direct and dynamic studies of specific Fg binding to activated platelets evaluated for native platelet-rich plasma, for washed platelets, and for activated, fixed platelets. We have demonstrated the equivalence of FITC-labelled and unlabelled Fg for binding to activated GPIIb-IIIa receptors, and in the rate and extent of mediating platelet aggregation. We found that FITC-Fg labelled at pH > or = 9 had reduced to absent specific binding to activated platelets, whether using soluble FITC or FITC-celite. The FITC-labelled Fg must be diluted 3-fold with unlabelled Fg when evaluating maximal Fg binding to activated platelets in order to prevent autoquenching of the FITC-Fg which leads to underestimation of Fg levels. The dissociation constant (KD) of Fg on stable preparations of activated, fixed platelets, determined with FITC-Fg binding to platelets by flow cytometry, was in the range reported for 125I-labelled Fg, 70-255 nm with Bmax = 10000-25000 Fg per platelet (n = 20). The FITC-Fg was used to monitor Fg binding to activated platelets directly by plasma, as well as to evaluate platelet subpopulations which maximally bind Fg according to the concentration of ADP used as activator. It is expected that this 'mildly' labelled FITC-Fg will stimulate further studies of platelet activation directly in native anticoagulated blood/plasma, for both basic and clinical research.
Blood Platelets↗
The tricyclic antidepressants clomipramine and citalopram induce apoptosis in cultured human lymphocytes.
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Antidepressive Agents, Tricyclic↗
Ultra low temperature cryopreservation of somatic embryogenic cell line of foxtail millet [Setaria italica (L.) Beauv].
Ultra low temperature cryopreservation is one of the methods for preservation of biological material. Until now, a major problem of protoplast culture of Gramineae is the instability of state of the somatic embryogenic cell line. In our experiments, elements affecting the ultra low temperature cryopreservation of somatic embryogenic cell line were studied: components of cryopreserve solution, somatic embryogenic cell line of different subculture time, growth recovery of cryopreserved cell line, and their protoplast cultures. Results demonstrated that the ultra low temperature cryopreservation did not change the properties of protoplast culture, and by using the cryopreserve method, plating efficiency of protoplast culture of cryopreserved cell line was maintained or enhanced.
Cell Line↗
[Chemical constituents of Gossampinus malabarica (L.) Merr. (II)].
Four chemical compounds isolated from the root of Gossampinus malabarica were identified as daucosterol, oleanolic acid, hesperidin and potassium nitrate by physico-chemical constants and spectroscopic analysis.
Drugs, Chinese Herbal↗
[Phospholipase A2 and multiple organ failure].
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Animals↗
Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis.
Apoptosis plays an important role during neuronal development, and defects in apoptosis may underlie various neurodegenerative disorders. To characterize molecular mechanisms that regulate neuronal apoptosis, the contributions to cell death of mitogen-activated protein (MAP) kinase family members, including ERK (extracellular signal-regulated kinase), JNK (c-JUN NH2-terminal protein kinase), and p38, were examined after withdrawal of nerve growth factor (NGF) from rat PC-12 pheochromocytoma cells. NGF withdrawal led to sustained activation of the JNK and p38 enzymes and inhibition of ERKs. The effects of dominant-interfering or constitutively activated forms of various components of the JNK-p38 and ERK signaling pathways demonstrated that activation of JNK and p38 and concurrent inhibition of ERK are critical for induction of apoptosis in these cells. Therefore, the dynamic balance between growth factor-activated ERK and stress-activated JNK-p38 pathways may be important in determining whether a cell survives or undergoes apoptosis.
Alkaloids↗
Bacterial expression of functional membrane-bound thromboxane synthase having intact sequence and truncated N-terminal hydrophobic segment.
A full-length cDNA for human placental thromboxane synthase and a shortened cDNA lacking the sequence corresponding to the N-terminal 2-29 amino acids were expressed in Escherichia coli using a pCW expression vector. Both intact and truncated recombinant enzyme were found in the membrane fraction and were catalytically active. These results suggest that the N-terminal hydrophobic segment, a proposed membrane anchor for P-450 enzymes, is not solely responsible for attachment of thromboxane synthase to the membrane and is not required for the proper protein folding or the enzyme activity.
Amino Acid Sequence↗