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Biomedical subjects

D Yuan

Publications and source records attributed to D Yuan.

At least 37 records · Page 2Linked to original sources

Recognition of multiple patterns of DNA sites by Drosophila homeodomain protein Bicoid.

Our previous studies demonstrated that the Drosophila homeodomain protein, Bicoid (Bcd), binds DNA cooperatively. In this study, we determined the patterns of adjacent DNA sites required for cooperative recognition by Bcd. Our in vitro selection and biochemical experiments demonstrated that Bcd binds preferentially to both head-to-head and tail-to-tail symmetric sites that are separated by short spacing. An increase in the spacing reduces the strict requirement of symmetric patterns of adjacent sites, permitting Bcd to recognize tandem repeat sites cooperatively. Our further experiments in vivo showed that the only pair of optimally spaced symmetric Bcd sites in a hunchback (hb) enhancer element contributes the most to transcriptional activation by Bcd, demonstrating the biological importance of the binding site patterns revealed by our in vitro selection studies.

Animals↗

Pharmacological properties of traditional medicines. XXV. Effects of ephedrine, amygdalin, glycyrrhizin, gypsum and their combinations on body temperature and body fluid.

Effects of ephedrine, amygdalin, glycyrrhizin, gypsum and their combinations on body temperature and body fluid were studied in rats using the method developed in our previous reports. Ephedrine significantly increased respiratory evaporative water loss and heat loss in response to a marked elevation of body temperature. There was a small but significant increase in body temperature when amygdalin was orally given rats at a dose of 46.32 mg/kg. Glycyrrhizin and gypsum were unable to affect body temperature. However, gypsum was able to prevent the increased action of ephedrine on body temperature, amygdalin exhibited a preventive tendency to it, and glycyrrhizin did not affect it. The results are in good agreement with classical claims of Makyo-kanseki-to and the related crude drugs in traditional medicine. Moreover, a combination of the four components reproduced the effects of Makyo-kanseki-to on body temperature and body fluid. This report suggests that the co-administration of ephedrine and gypsum is physiologically more desirable than ephedrine alone for dry-type asthmatic patients with a fever. Also, it experimentally supports the clinical efficacy of Makyo-kanseki-to.

Amygdalin↗

Comparison of the mutagenicity of exhaust emissions from motor vehicles using leaded and unleaded gasoline as fuel.

While unleaded gasoline has the advantage of eliminating lead from automobile exhaust, its potential to reduce the exhaust gas and particles, merits further examination. In the present studies, the concentrations of hydrocarbons (HC) and carbon mono-oxides (CO) in emissions were analyzed on Santana engine Dynamometer under a standard test cycle, and total exhaust particles were collected from engines using leaded and unleaded gasoline. It was found that unleaded gasoline reduced the emissions of CO and HC, and decreased the quantity of vehicle exhaust particulate matters by 60%. With the unleaded gasoline, only 23 kinds of organic substances, adsorbed in the particles, were identified by gas chromatography/mass spectrometer (GC/MS) while 32 components were detected using the leaded gasoline. The results of in vitro Salmonella/microsomal test and micronucleus induction assay in CHL cells indicated that both types of gasoline increased the number of histidine-independent colonies and the frequencies of micronucleus induction; no significant difference was found in their mutagenicity.

Carbon Monoxide↗

Effect of upstream RNA processing on selection of mu S versus mu M poly(A) sites.

All of the regulatory factors responsible for augmenting microseconds mRNA levels preceding the dramatic increase in secretory IgM production upon B cell activation has not been totally elucidated. Whereas previous experiments have centered on the region of the gene specifying the choice between splicing to mu M exons versus selection of the mu S poly(A) site, we have found that upstream sequences within the Cmu gene, specifically the Cmu 4 acceptor splice site together with intronic sequences between the Cmu 3++ and Cmu 4 exons, play an important role in dictating the precision or the extent of splicing to the mu M exons even under conditions in which functional polyadenylation factors should be in excess. Therefore, splicing of upstream exons can affect remotely located downstream exons. These findings suggest that regulation of differential mu S/mu M mRNA expression may involve general processing enzymes that recognize specific cis -regulatory sequences residing within the body of the mu gene and account for the unique ability of activated B cells to secrete copious amounts of IgM.

Alternative Splicing↗

Pharmacological properties of traditional medicines. XXIV. Classification of antiasthmatics based on constitutional predispositions.

It is commonly accepted that bronchial asthma or rhinitis accompanies disorders in body fluids and body temperature. The effects of ephedrine and the traditional antiasthmatics "Makyo-kanseki-to" and "Goko-to" were therefore studied on such constitutional predispositions as insensible perspiration and body temperature. Ephedrine markedly increased body temperature and exhibited a strong increased action on respiratory insensible perspiration, whereas Makyo-kanseki-to and Goko-to not only prevented the elevation of body temperature, but also increased respiratory insensible perspiration following the reduction of non-evaporative heat loss from the body surface. Thus, the diagnostic criteria of these two medicines used to treat hot-type asthma or dry cough were experimentally determined. The results also suggest that there is a great possibility that the administration of antiasthmatics may elicit side effects or make diseases worse unless their actions on constitutional predispositions are taken into account, such as body temperature and body fluids.

Amygdalin↗

A new experimental device for the measurement of moisture emission and heat release from respiratory organs and body surface.

A new experimental device was developed to investigate respiratory diseases. The moisture and heat released from respiratory organs and the body surface of a rat were determined by means of this device as well as the rectal temperature. The high recovery of results was statistically confirmed, and the measured values at various environmental temperatures were significantly different from each other. Some standard drugs, such as ephedrine, aminophylline and chlorpromazine, were examined. Their stimulant or depressant actions were clearly observed. The results of some traditional medicines for the treatment of rhinitis and bronchial asthma from this measuring system were consistent with their clinical applications. These results suggest that this new experimental system is not only effective in the experimental understanding of cold-hot syndrome, but also contributes to the evaluation of the effects of traditional medicines.

Animals↗

The effect of NK cell activation by tumor cells on antigen-specific antibody responses.

In addition to mediating direct cytotoxicity, NK cells can exert regulatory effects on specific immune responses. For example, injection of poly (I:C) can alter specific Ab responses, which is attributable to the production of IFN-gamma by NK cells. To test whether direct activation of NK cells can exert the same effect, we have injected, at the same time as Ag challenge, BCL1-C11 tumor cells, which are highly effective inducers of IFN-gamma production by NK cells. The results show a specific enhancement of the IgG2a response, which does not occur with a tumor (70Z/3) that does not induce IFN-gamma production. This enhancement is NK cell and IL-12 dependent. However, BCL1-C11 cells cannot directly induce IL-12 production in peritoneal exudate cells (PECs). On the other hand, PECs from tumor-treated mice produce IL-12 in response to LPS, suggesting that they are primed in vivo. Furthermore, the IL-12 production is NK cell and IFN-gamma dependent. These results indicate that if tumor cells can directly activate NK cells to produce IFN-gamma, this cytokine initiates an amplification loop by activating macrophages to produce IL-12, which in turn activates NK cells further, resulting in the alteration of the isotype distribution of specific Ab responses. Production of the appropriate Ab isotype should enhance Ab-dependent cellular cytotoxicity against targets mediated by NK cells, implicating their role(s) in the specific immune response as well as the initial nonspecific phase.

Animals↗

Effect of the presence of transgenic H and L chain genes on B cell development and allelic exclusion.

The expression of various components of the antigen receptor is closely associated with the developmental progression of B lymphocytes. For this reason, introduction of rearranged Ig transgenes exerts profound effects on B cell development. In these studies we show that the presence of transgenes for both mu and delta H as well as L chains accelerates the rate of B cell maturation resulting in the appearance of large numbers of IgD-expressing B lymphocytes in vitro that are otherwise undetectable. In addition, allelic exclusion, normally exerted very effectively by these particular transgenes (carried in the MD-3 strain), was lost in the long-term bone marrow cultures. These findings can be recapitulated in vivo by serial adoptive transfers of bone marrow cells from the transgenic animals into lethally irradiated recipients. We conclude from these studies that allelic exclusion is not necessarily mediated by any one event but may be a result of the integrated occurrence of Ig H chain gene rearrangement, accumulation of transcription factors, along with the ordered progression of B cell differentiation under the influence of the appropriate inductive microenvironment. These findings may account for at least some of the observed cases of allelic inclusion in transgenic animals.

Alleles↗

[Clinial study of "qingjunyin" detoxification for the treatment of heroin addicts].

To evaluate the efficacy of treatment of heroin addicts (n = 100) by "Qingjunyin" (QJY) detoxification (10 days program). Methadone detoxification (10 days program) group (n = 50) and colonidine treated group (n = 50) serve as controls. QJY group is given 180 ml P. O daily. The dosage of the two comparatine groups is given according to Documentation, the method of abserving the treatment protocols and detoxification standard is according to current regulation. The scores of abstinence syndrome in QJY detoxification group are lower than those in colonidine treated group in the first three days of potocol, but this difference disappear in the late stage of treatment. While QJY detoxification is effective as methadone detoxification in the control of abstinece syndrome during the first five days of treatment but the difference in the scores of abstinence syndrome between QJY and methadone group is observed during the late five days of protocol. QJY does not result in potential dependence and has definite curative effect in the treatment of heroin addiction.

Adolescent↗

Regulation of IgM and IgD heavy chain gene expression: effect of abrogation of intergenic transcriptional termination.

Early IgM+ B cells express little or no membrane IgD due to the low abundance of delta mRNA. Extensive transcriptional termination regulated by sequences in the intronic region between mu and delta heavy chain genes may be the primary reason for the lack of delta gene transcription. We have examined the effect of deletion of these sequences on the regulation of IgM and IgD heavy chain gene expression in transfectants as well as mice carrying this otherwise intact transgene. By run-on transcriptional measurement, we show that the delta exons are transcribed in bone marrow B cells from these transgenic mice. However, in spite of the induced premature synthesis of the full-length mu-delta transcript in pre-B cells, processing to delta mRNA does not occur until the lymphocytes express cell surface IgM. Therefore, during B cell development, synthesis of the full-length transcript is a necessary but not sufficient condition for initiation of delta mRNA synthesis. Furthermore, unexpectedly, the abrogation of transcriptional termination was found to also affect the processing of the primary transcript to microM mRNA. These results show that expression of IgD in primary B cells is stringently regulated and closely linked to IgM expression.

Animals↗

Sequences outside the homeodomain of bicoid are required for protein-protein interaction.

The Drosophila morphogenetic protein Bicoid (Bcd) is required for the development of anterior structures of the embryo. Bcd, a homeodomain protein, is distributed as an anterior-to-posterior gradient in the embryo. It stimulates the expression of the hunchback (hb) gene in the anterior half in an all-or-none fashion. We have recently shown that Bcd binds cooperatively to a hb enhancer element and proposed that cooperative DNA binding is facilitated by an interaction between Bcd molecules. In this report, we further analyze the interaction between Bcd molecules and define regions important for protein-protein interaction. We show that the homeodomain of Bcd alone fails to interact with another Bcd molecule efficiently. The protein sequence flanking either side of the homeodomain restores the protein-protein interaction function. Mutations in the homeodomain that affect DNA binding do not adversely affect the protein-protein interaction function, suggesting that the surfaces for DNA binding and protein-protein interaction are separable. Finally, we demonstrate that the homeodomain of Bcd alone, unlike the intact Bcd, fails to bind DNA cooperatively. These results further support the notion that cooperative DNA binding is facilitated by the interaction between Bcd molecules. They strongly suggest that protein-protein interaction is an important property of Bcd for its biological activities.

DNA Footprinting↗

The role of NK cells during in vivo antigen-specific antibody responses.

Investigations into the role of NK cells in regulating Ab responses have yielded variable results, some suggesting that NK cells can down-regulate Ag-specific Ig production and others proposing an enhancing effect. These apparently inconsistent findings may stem partially from the specificity of reagents used in purifying cell populations and/or the nature of the in vitro systems used to study these events. We chose to investigate the ability of either resting or poly(I:C)-activated NK cells to alter an in vivo Ab response in mice given a T-independent (TNP-LPS) or T-dependent (TNP-keyhole limpet hemocyanin (KLH)) Ag. By using a more specific Ab, anti-NK-1.1, to deplete NK cells, we were able to clearly show that resting, endogenous NK cells do not affect either type of response, as measured by serum Ag-specific Ig levels quantitated by isotype-specific ELISA. In contrast, activation of NK cells by poly(I:C) increased Ag-specific IgC2a as well as IgG1 levels. Interestingly, only the effect on IgG2a production is reversible by depletion of NK cells.

Animals↗

The Drosophila morphogenetic protein Bicoid binds DNA cooperatively.

The Drosophila morphogenetic protein Bicoid, encoded by the maternal gene bicoid, is required for the development of the anterior structures in the embryo. Bicoid, a transcriptional activator containing a homeodomain, is distributed in an anterior-to-posterior gradient in the embryo. In response to this gradient, the zygotic gene hunchback is expressed uniformly in the anterior half of the embryo in a nearly all-or-none manner. In this report we demonstrate that a recombinant Bicoid protein binds cooperatively to its sites within a hunchback enhancer element. A less than 4-fold increase in Bicoid concentration is sufficient to achieve an unbound/bound transition in DNA binding. Using various biochemical and genetic methods we further demonstrate that Bicoid molecules can interact with each other. Our results are consistent with previous studies performed in the embryo, and they suggest that one mechanism to achieve a sharp on/off switch of gene expression in response to a morphogenetic gradient is cooperative DNA binding facilitated by protein-protein interaction.

Animals↗

[Effect of HE-NE laser acupuncture on the spleen in rats].

The splenic sinuses, lymphocytes and antigen presenting cells in the spleen of rat stimulated by He-Ne laser acupuncture were observed by using TEM to investigate the ultrastructural changes of them. The width of endothelial interstice was increased, hence a lot of blood cells oozed out of the splenic sinuses. There were numerous activated T cells which contained abundant mitochondria and ribosome in the periarterial lymphatic sheaths. The B cells were gradually differentiated into large lymphocytes, immature and mature plasmatic cells with a lot of endoplasmic reticula. They were prominently increased in the splenic nodules. The macrophages had short processes with numerous folds and microvilli and tended to neighboring lymphocytes. The nucleus pores were increased. There were a lot of vacuoles, phagosome and mitochondria were closed to macrophages and lymphocytes. In conclusion the activities of the cellular immunity and humoral immunity were enhanced by laser.

Acupuncture Points↗

Vulnerability of the hippocampus to kainate excitotoxicity in the aged, mature and young adult rat.

Sensitivity to excitotoxic damage was assessed in young adult, mature and aged male Sprague-Dawley rats. Kainic acid was injected into the hippocampus and the size of the hippocampal lesion rated. Intrahippocampal injection of kainic acid produced lesions in aged animals that were significantly smaller than lesions in the young rats (P < 0.05), while lesion size in mature rats was intermediate. Excitotoxic damage was localized primarily to the CA3 region of the hippocampus in the aged rats. Young adult rats had more damage to the hippocampus with involvement of CA1 pyramidal and dentate granule cells. These results suggest that increased age may reduce susceptibility to excitotoxic damage.

Aging↗

Regulation of IFN-gamma mRNA production in murine natural killer cells.

We have previously reported that large, presumably in vivo activated, B cells stimulate murine natural killer (NK) cells to secrete increased levels of IFN-gamma. In order to further understand the mechanism of IFN-gamma induction, we compared the regulation of IFN-gamma mRNA production after stimulation of NK cells with either B lymphocytes or phorbol myristate acetate (PMA)+ionomycin. Here we show that stimulation of NK cells by either stimuli results increase in IFN-gamma mRNA, albeit with different kinetics. Although the induction requires new RNA synthesis, we could not detect increased transcription of the IFN-gamma gene after stimulation. Measurement of the rate of mRNA degradation after IFN-gamma mRNA has accumulated demonstrates that this mRNA is more stable than IFN-gamma mRNA from unstimulated NK cells. Together, these results suggest that the increase in IFN-gamma mRNA and protein in NK cells, stimulated by B cells or PMA+ionomycin, results from stabilization of pre-existing IFN-gamma message. Our results also suggest that induction of the factor which stabilizes the mRNA, although as yet unknown, requires new RNA synthesis.

Animals↗

Role of the OCTA site in regulation of IgH chain gene transcription during B cell activation.

We have evaluated the importance of the OCTA site in both the promoter and enhancer regions for the induction of enhancement of IgH chain gene transcription after B cell activation. These studies show that although occupancy of the OCTA site in the promoter is critical for basal transcription of the mu gene, it is not necessary for the increase in transcription induced by in vivo activation. On the other hand, the OCTA site in the enhancer is necessary for neither basal transcription nor in vivo activation of transcription; however, occupancy of this site is required for further up-regulation in transcription of the mu gene in pre-activated cells. These results indicate that different mechanisms may be involved in the activation of resting versus in vivo stimulated B lymphocytes. The findings are discussed in relation to the phenotype described for Oct-2-deficient mice.

Animals↗