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Biomedical subjects

J Jiang

Publications and source records attributed to J Jiang.

At least 415 records · Page 23Linked to original sources

Stable expression of the rat GLP-I receptor in CHO cells: activation and binding characteristics utilizing GLP-I(7-36)-amide, oxyntomodulin, exendin-4, and exendin(9-39).

Glucagon-like peptide-I (GLP-I) is a potent insulinotropic peptide that mediates its actions at pancreatic B-cells via specific receptors. In the present study we stably expressed the rat B-cell GLP-I receptor in CHO cells and studied binding characteristics and receptor activation utilizing the naturally occurring receptor agonist GLP-I(7-36)-amide (GLP-I), the proglucagon-derived GLP-I-related peptide oxyntomodulin, the GLP-I receptor agonist exendin-4, and the specific antagonist exendin(9-39). The potencies to displace [125I]GLP-I from the receptor were GLP-I > exendin-4 > exendin(9-39) > oxyntomodulin, and to displace [125I]exendin-4 GLP-I = exendin-4 > exendin(9-39) > oxyntomodulin. cAMP production was stimulated equally by GLP-I and exendin-4. Oxyntomodulin was less potent to stimulate cAMP generation. Exendin(9-39) blocked the stimulatory action of GLP-I and exendin-4 on cAMP production, but not that of oxyntomodulin. This study shows that GLP-I and exendin-4 are potent agonists at the transfected rat B-cell GLP-I receptor whereas oxyntomodulin is only a weak GLP-I receptor agonist. Furthermore, exendin(9-39) is a potent GLP-I receptor antagonist. This peptide is a valuable tool to further study the physiological actions of GLP-I.

Animals↗

Bioconcentration of cadmium in water hyacinth (Eichhornia crassipes) in relation to thiol group content.

To study the bioconcentration of cadmium in water hyacinth, the plants were exposed to water containing 2 microg Cd2+/ml for extended periods of time. Three strains from several exposures during a 30-day period were sampled for the analyses of cadmium and thiol group. The data showed that the plant concentrates cadmium mainly in the roots and that the cadmium uptake is proportional to the increase of the thiol group content. The latter suggests the possibility of using the thiol group content to assess the bioconcentration of heavy metal ions in water hyacinth and as a general parameter for monitoring the heavy metal pollution of water. A simple two-compartmental model was used to simulate the kinetics of cadmium uptake. The calculated bioconcentration factor matches the one derived directly from experimental data, indicating the adequacy of the model.

Journal Article↗

In vivo measurement of oxygen concentration using sonochemically synthesized microspheres.

Proteinaceous microspheres filled with nitroxides dissolved in an organic liquid have been synthesized for the first time using high intensity ultrasound; these were used to measure oxygen concentrations in living biological systems. The microspheres have an average size of 2.5 microns, and the proteinaceous shell is permeable to oxygen. Encapsulation of the nitroxides into the microsphere greatly increased the sensitivity of the electron paramagnetic resonance signal line width to oxygen because of the higher solubility of oxygen in organic solvents. The encapsulation also protected the nitroxide from bioreduction. No decrease in intensity of the electron paramagnetic resonance signal was observed during 70 min after intravenous injection of the microspheres into a mouse. Measurement of the changes in oxygen concentration in vivo by means of restriction of blood flow, anesthesia, and change of oxygen content in the respired gas were made using these microspheres.

Anesthetics, General↗

Doxorubicin-induced changes in intracellular Ca2+ transients observed in cardiac myocytes isolated from guinea-pig heart.

Effects of doxorubicin on intracellular Ca2+ concentrations ([Ca2+]i) were examined using myocytes isolated from guinea-pig heart and loaded with fura 2. Changes in twitch contractions were assessed from the motion of myocytes. Ca2+ transients and contractions were triggered by electrical-field stimulation. Exposure of myocytes to doxorubicin depressed Ca2+ transients and contractions. The time to peak Ca2+ transient was prolonged, and Ca2+ sequestration was delayed. In myocytes treated with doxorubicin, an increase in external CaCl2 concentration from 1.2 to 3.6 mM increased the resting and peak [Ca2+]i and enhanced twitch contraction. In the presence of 3.6 mM CaCl2, isoproterenol failed to enhance Ca2+ transients or contractions of doxorubicin-treated myocytes. Effects of doxorubicin were compared with those of agents known to alter Ca2+ handling by myocytes. Caffeine enhanced the peak and resting [Ca2+]2 and contraction. Verapamil caused a rapid decrease in Ca2+ transients and twitch contractions. The effects of verapamil were reversed by isoproterenol in the presence of 3.6 mM CaCl2. Ryanodine alone and combined with doxorubicin depressed contractions and Ca2+ transients and elevated resting [Ca2+]i. Resting [Ca2+]i was further elevated by an increase in CaCl2 concentration and the addition of isoproterenol. The combination of verapamil and ryanodine inhibited Ca2+ transients and contractions. In the presence of verapamil and ryanodine, an increase in extracellular CaCl2 concentration increased resting and peak [Ca2+]i. Isoproterenol further elevated resting and peak [Ca2+]i and increased twitch contractions. These results indicate that doxorubicin alters cellular Ca2+ handling. The actions of doxorubicin are not mimicked by caffeine, verapamil, ryanodine, or the combination of verapamil and ryanodine. Among these agents, ryanodine produced effects that were closest to those observed with doxorubicin.

Animals↗

Detection of the human glucagon-like peptide 1(7-36) amide receptor on insulinoma-derived cell membranes.

125I-glucagon-like peptide 1(7-36)amide was covalently cross-linked to a specific binding protein in human insulinoma cell membranes. A single radiolabeled band at M(r) 63,000 was identified by SDS-PAGE after solubilization of the ligand-binding protein complex. The molecular weight of this apparent GLP-1 receptor in human endocrine pancreatic tissue was of identical size as the GLP-1 receptor on rat insulinoma-derived RINm5F cell membranes. The radiolabeled band was undetectable when 1 microM of unlabeled GLP-1(7-36)amide or of the GLP-1 antagonist exendin(9-39)amide was included in the binding assay. Utilizing isolated poly-A+ RNA from the human insulinoma and a 1,500 bp Eco-RI fragment of the cDNA coding for the rat GLP-1(7-36)amide receptor for Northern blot analysis, a main hybridization signal at about 7 kb was found by Northern blotting. Our data provide the first direct evidence of the existence of GLP-1 receptors in human endocrine pancreatic tissue.

Animals↗

[Therapeutic effectiveness of zuzhongping on patients with arteriosclerostic cerebral infarction].

Forty-six patients with acute arteriosclerostic cerebral infarction were randomly divided into two groups: control group and treatment group. Each of them included 23 patients respectively. The patients in the control group were given Dextran-40 but the ones in the treatment group were given the mixture of Zuzhongping. The course of treatment was 3 weeks. It was found that there was a significant difference (P < 0.01) in the score percentage, before and after treatment of neurological defects, between the control group and the treatment group, and the former (29.70 +/- 33.52) was much lower than the latter (45.40 +/- 27.60). The total curative rate of the treatment group (87.0%, 20/23) was significantly higher than that of the control group (60.9%, 14/23). There was an obviously prolonged KPTT (kaolin partial thromboplastin time) value and a decreased Fb (fibrinogen) level in the treatment group. Before treatment they were 32.43 +/- 4.03 sec and 6.18 +/- 1.77 g/L respectively, but after treatment, 52.96 +/- 10.50 sec and 4.5 +/- 0.95 g/L respectively. The authors suggest that the significant therapeutic efficacy of Zuzhongping in the patients with acute arteriosclerostic cerebral infarction is related to its action of anticoagulation, modification of PGI2 and TXA2 level in the body, decreased blood Fb level, hyperglycemia, etc.

Aged↗

[A primary study of the factors related to the patients' adherence of anti-hypertensive therapy].

An interview covering demographic information, history of hypertension as well as medical and health-related questions was conducted on 540 hypertension patients in Chengdu. The objective of the study was to analyse the factors related to the adherence to antihypertensive therapy of these patients. The simple analysis showed that the education level of the patients, the type of payment for visiting physician, the convenience of visiting physician, the awareness of the severity of hypertension, the average family income of the patients were related to the adherence to therapy. The step multi-regression analysis showed that the type of payment for visiting physician, the convenience of visiting physician and the average family income were related to the adherence to antihypertensive therapy.

Adult↗

[Changes of norepinephrine release in rat's nucleus reticularis paragigantocellularis lateralis in acupuncture analgesia].

Norepinephrine (NE) of the nucleus reticularis paragigantocellularis lateralis (RPGL) plays an important role in acupuncture analgesia (AA). The aim of this experiment was to investigate the role of NE in RPGL during AA at presynaptic level by using push-pull perfusion, high performance liquid chromatography with electrochemical detection (HPLC-ECD) techniques. The results indicated that pain threshold increased significantly after 20 min of EA, while the content of NE and its metabolite MHPG in the perfusate from the RPGL was markedly decreased. A negative correlation existed between the changes of pain threshold and the release of NE and MHPG. It is indicated that electroacupuncture (EA) is able to inhibit the release of NE from the RPGL during AA.

Acupuncture Analgesia↗

[Cloning and sequencing of the major surface antigen gene of Toxoplasma gondii].

According to the published gene sequence of the major surface antigen (P30), a pair of primers were designed and synthesized. At the 5' end of sense and anti-sense strand of the primers, EcoRI and BamHI sites were added, respectively. Using PCR, the coding sequences of P30 gene were amplified. The amplified gene fragments and plasmid pBV220 were digested with EcoRI and BamHI, and then ligated. The inserted gene fragment was sequenced by the chain termination method. The reading revealed that nucleotide sequence determined here was the same as the sequence published by Burg (1988), except one base was exchanged. The recombinant plasmid pBV220-P30 was constructed.

Amino Acid Sequence↗

Ligand-specificity of the rat GLP-I receptor recombinantly expressed in Chinese hamster ovary (CHO-) cells.

Glucagon-like peptide-I (GLP-I) is a potent incretin hormone and is considered as a new therapeutic tool in the treatment of diabetes mellitus. This study was designed to precisely characterize the binding behavior and activation of the recombinant GLP-I receptor against naturally occurring ligands of the glucagon/VIP/secretin peptide hormone family. CHO-cells were stably transfected with a plasmid containing a cDNA encoding for the rat GLP-I receptor. Northern blot analysis with this cDNA showed a single band of 2.7 kb in CHO cells, while in RINm5F cells, three bands of 2.7, 3.4, and 3.6 kb were specifically labelled. In receptor-binding studies 125I-GLP-I was displaced by GLP-I and weakly by PHI and oxyntomodulin but not by helodermin, helospectin I, helospectin II, secretin, VIP, and PACAP-38. Intracellular cAMP generation was stimulated by GLP-I, PHI, and oxyntomodulin. Helodermin, helospectin I, helospectin II, secretin, VIP, and PACAP-38 were not able to displace 125I-GLP-I from its receptor or to stimulate intracellular cAMP production. This data shows that the GLP-I receptor is characterized by a high ligand specificity.

Animals↗

Binding affinities and cooperative interactions with bHLH activators delimit threshold responses to the dorsal gradient morphogen.

The dorsal (dl) morphogen gradient initiates the formation of the mesoderm, neuroectoderm, and dorsal ectoderm by setting different limits of regulatory gene expression along the dorsoventral axis of the early Drosophila embryo. In this paper, we show that low affinity dl-binding sites restrict target gene expression to the ventralmost regions (presumptive mesoderm), where there are peak levels of dl, while high affinity sites permit expression in ventrolateral regions (mesoderm and mesectoderm) containing intermediate levels of the morphogen. Activation by low levels of dl in lateral regions (the presumptive neuroectoderm) depends on cooperative DNA binding interactions between dl and bHLH proteins. The snail repressor blocks this interaction and restricts expression to the neuroectoderm. We discuss how enhancers serve as templates to bring weakly interacting regulatory factors into close proximity so that they can function combinatorially to activate and repress transcription.

Animals↗

Protein kinase C: a novel target for inhibiting gastric cancer cell invasion.

BACKGROUND: Gastric adenocarcinoma is a common neoplasm worldwide. Patients with completely resected disease often have locoregional recurrence, and adjuvant chemotherapy has failed to reduce the common occurrence of metastases. Protein kinase C (PKC) is thought to be important in tumor cell invasion, but its relationship to gastric cancer cell invasion, and thus metastases, remains unexplored. We recently identified and established invasive and noninvasive human gastric adenocarcinoma cell lines, which can now be used to test agents for inhibition of tumor cell invasion by inhibition of PKC activity. PURPOSE: The objectives were (a) to test threo-dihydrosphingosine (SPC100221), a specific inhibitor of PKC at its regulatory site, and staurosporine, a potent but nonspecific inhibitor of PKC at its catalytic site, for their effects on gastric cancer cell invasion in vitro and (b) to determine whether the expression of PKC isoforms can distinguish invasive from noninvasive gastric cancer cells. METHODS: Gastric cancer cell invasion through Matrigel-coated Nuclepore filters in the Boyden chamber assay was analyzed in the presence of graded concentrations of SPC100221 and staurosporine. The invasive SK-GT-1 and SK-GT-5 cell lines and the noninvasive SK-GT-2 and SK-GT-4 cell lines were used. PKC isoform expression was determined by reverse transcription of messenger RNAs to complementary DNA and subsequent amplification by the polymerase chain reaction. RESULTS: The effects of staurosporine and SPC100221 on tumor cell invasion were tested at drug concentrations that did not inhibit cell proliferation, as evidenced by [3H]thymidine uptake. Staurosporine and SPC100221 at subtoxic doses inhibited human gastric cancer cell invasion by 50% at 5 x 10(-9) M and 2 x 10(-7) M, respectively. The expression of PKC beta was observed in the invasive but not the noninvasive gastric cancer cells. Both types of cells, however, expressed the PKC alpha and PKC gamma isoforms. CONCLUSIONS: Gastric cancer cell invasion can be inhibited by PKC inhibitors, and expression of PKC beta may be a marker of invasiveness in gastric cancer. IMPLICATIONS: PKC appears to represent a new target for inhibition of gastric cancer cell invasion, and SPC100221, in view of its PKC specificity, may provide a model for future drug development in this area. Moreover, PKC beta may have a fundamental role in the development of invasive potential in gastric cancer.

Alkaloids↗

Conversion of a dorsal-dependent silencer into an enhancer: evidence for dorsal corepressors.

In Drosophila, the dorsal (dl) morphogen gradient initiates the differentiation of the embryonic mesoderm and neuroectoderm by activating the expression of regulatory genes (e.g. twist and snail) in a concentration-dependent manner. dl also functions as a repressor that establishes the dorsal epidermis and amnioserosa by restricting regulatory genes such as dpp and zen to dorsal regions of the embryo. The ability of dl to function as both an activator and repressor distinguishes it from the bicoid morphogen, which appears to function solely as an activator. In an effort to determine how dl functions as a repressor we have performed a detailed characterization of a zen silencer element, called the VRE, which mediates ventral repression in response to the dl gradient. A minimal 110 bp VRE sequence is identified, which is able to silence the ventral expression of a heterologous promoter. This sequence contains two dl binding sites as well as binding sites for additional nuclear factors present in early embryos. Mutations in the latter binding sites convert the minimal VRE into an enhancer, which mediates transcriptional activation in ventral regions in response to dl. These results suggest that dl is intrinsically an activator, but is converted into a potent silencer when it interacts with neighboring corepressors.

Animals↗

A 'zebra' chromosome arising from multiple translocations involving non-homologous chromosomes.

An alloplasmic wheat line carrying a 'zebra' chromosome z5A was isolated from the derivatives of an Elymus trachycaulus x Triticum aestivum cv Chinese Spring hybrid. Chromosome z5A was named zebra because of its striped genomic in situ hybridization pattern. z5A consists of four chromosome segments derived from E. trachycaulus and four chromosome segments, including the centromere, from wheat. The short arm of z5A paired with the telocentric chromosome 1H(t)S of E. trachycaulus and the long arm with the long arm of normal 5A. z5A also carried several genetic markers derived from 1H(t)S. Chromosome 1H(t) was the only E. trachycaulus chromosome found in the sib plants of a previous generation from which z5A was derived. Monosomic 5A and telocentric chromosome 5AL were also found in most of the sib plants. The zebra chromosome most probably originated from spontaneous multiple translocations between chromosomes 5A and 1H(t)S or 5A and 1H(t).

Chromosome Banding↗

Induction of an adaptive response to dominant lethality and to chromosome damage of mouse germ cells by low dose radiation.

In the present paper, dominant lethal mutations, chromosome aberrations in spermatocytes and reciprocal translocations in stem spermatogonia were analyzed after whole body exposure of mice to X-radiation. Results both from chromosome aberrations in spermatocytes and for reciprocal translocations in spermatogonia showed that pre-exposure to low doses up to 200 mGy could induce a significant dose-dependent reduction in adapted mice compared to the non-adapted mice; the lower the adaptive dose, the greater the reduction. For dominant lethal mutations, it was found that spermatogonia (both stem cells and differentiated cells) and spermatocytes adapted to 50 mGy X-rays could show an adaptive response, but spermatids and spermatozoa could not.

Adaptation, Physiological↗

Modulated excitation of singly ligated carboxyhemoglobin.

We have extended the method of modulated excitation, a small perturbation kinetic method, to study ligand binding and conformational change of hemoglobin tetramers with a single ligand bound. To restrict the excitation to the first ligand, only 1% of the hemes have bound CO, and the remainder are kept unliganded. A detailed theory is presented which agrees well with the experimental observations. This method of observing ligand recombination also provides a novel and simple method for determination of hemoglobin concentration. Additional relaxation processes are also observed. By fitting independently determined spectra to the spectra associated with the relaxations, these processes are assigned as thermal excitation and thermally driven protonation/deprotonation reactions. These added relaxations arise from the deoxy-Hb portion of the samples, and demonstrate that modulated excitation can be used effectively for temperature perturbation in the absence of photodissociation. The spectra observed are not well described by the spectra of allosteric change, however, and we conclude that there is no significant mixing of quaternary states at the first ligation step. In an appendix we present a derivation of the particular features seen in thermally modulated protonation reactions.

Allosteric Site↗

Allosteric kinetics and equilibria of triligated, cross-linked hemoglobin.

Using modulated excitation, we have measured the forward and reverse rates of the allosteric transition between relaxed (R) and tense (T) quaternary structures for triply ligated hemoglobin (Hb), cross-linked between the alpha chains at Lys 99. Oxygen, carbon monoxide, and water were used as ligands and were studied in phosphate and low Cl- bis-Tris buffers at neutral pH. Since the cross-link prohibits disproportionation, triply ligated aquomet Hb species with ferrous beta chains were specifically isolated by isoelectric focusing. Modulated excitation provides rate pairs and therefore gives equilibrium constants between quaternary structures. To coordinate with that information, oxygen binding curves of fully ferrous and tri-aquomet Hb were also measured. L3, the equilibrium constant between three liganded R and T structures, is determined by modulated excitation to be of order unity for O2 or CO (1.1 to 1.5 for 3O2 and 0.7 for 3CO bound), while with three aquomet subunits it is much greater (> or = 23). R-->T conversion rates are similar to those found for HbA, with weak sensitivity to changes in L3. The L3 values from HbXL O2 were used to obtain a unique allosteric decomposition of the ferrous O2 binding curve in terms of KT, KR, and L3. From these values and the O2 binding curve of tri-aquomet HbXL, L3 was calculated to be 2.7 for the tri-aquomet derivative. Consistency in L3 values between equilibrium and modulated excitation data for tri-aquomet-HbXL can be achieved if the equilibrium constant for O2 binding to the alpha chains is six times lower than that for binding to the beta chains in the R state, while the cooperative properties remain homogeneous. The results are in quantitative agreement with other studies, and suggest that the principal effect of the cross-link is to decrease the R state and T state affinity of the alpha subunits with almost no change in the affinity of the beta subunits, leaving the allosteric parameters L and c unchanged.

Allosteric Regulation↗