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Biomedical subjects

C Gao

Publications and source records attributed to C Gao.

At least 19 recordsLinked to original sources

Slow excitatory synaptic transmission mediated by P2Y1 receptors in the guinea-pig enteric nervous system.

Electrophysiological recording was used to study a type of slow excitatory postsynaptic potential (slow EPSP) that was mediated by release of ATP and its action at P2Y1 receptors on morphologically identified neurones in the submucosal plexus of guinea-pig small intestine. MRS2179, a selective P2Y1 purinergic receptor antagonist, blocked both the slow EPSP and mimicry of the EPSP by exogenously applied ATP. Increased conductance accounted for the depolarization phase of the EPSP, which occurred exclusively in neurones with S-type electrophysiological behaviour and uniaxonal morphology. The purinergic excitatory input to the submucosal neurones came from neighbouring neurones in the same plexus, from neurones in the myenteric plexus and from sympathetic postganglionic neurones. ATP-mediated EPSPs occurred coincident with fast nicotinic synaptic potentials evoked by the myenteric projections and with noradrenergic IPSPs evoked by sympathetic fibres that innervated the same neurones. The P2Y1 receptor on the neurones was identified as a metabotropic receptor linked to activation of phospholipase C, synthesis of inositol 1,4,5-trisphosphate and mobilization of Ca2+ from intracellular stores.

Adenosine Diphosphate↗

Non-filamentous sludge bulking caused by a deficiency of nitrogen in industrial wastewater treatment.

Deficiency in the nutrient supply such as nitrogen usually results in activated sludge bulking and this phenomenon often takes place in the industrial wastewater treatment plants with activated sludge process. The effects of nitrogen deficiency on activated sludge bulking were studied specially in some experiments carried out in a sequencing batch reactor fed with brewing process wastewater in this paper. The experimental results showed that the sludge settled properly at an influent BOD/N value of 100/4. When the value of BOD/N was 100/3, filaments had an excessive growth at one time during the reaction process. Afterwards, the number of filamentous bacteria began to reduce and simultaneously an excessive growth of viscous Zoogloea with high percentage of moisture was observed and non-filamentous activated sludge bulking occurred. When the influent BOD/N value was 100/2, the excessive growth of filamentous microorganisms could not be observed at all times and the sludge characterization was similar to the case in which BOD/N value was 100/3. When the value of influent BOD/N was 100/0.94, more serious non-filamentous bulking occurred. Furthermore, the effects of nitrogen deficiency on the nitrogen sources and phosphorus sources utilization rate and the COD removal rate were investigated in the experiments.

Beer↗

A second look at anorectal infections in cancer patients in a large cancer institute: the success of early intervention with antibiotics and surgery.

BACKGROUND: Infection of anorectal region represents a significant complication of anti-cancer therapy. Anorectal infection occurs in patients receiving aggressive chemotherapy. Untreated infection leads to substantial morbidity and in the past, mortality. METHODS: 82 episodes of anorectal infection in 64 patients with malignant diseases occurring over 12 years at the National Cancer Institute (NCI) were retrospectively reviewed. RESULTS: The overall incidence is comparable to the prior NCI experience despite a shift in patient population to a lower percentage of lymphoid/leukemic diagnoses (34% vs 77%). There were no deaths associated with anorectal infection in the 12 years reviewed compared to seven of 44 in the previous decade (p = 0.003). Antibiotic therapy alone was successful in managing 25/82. Only five episodes were treated with surgery alone compared to nearly 45% in the previous decade. There were no major surgical complications. Neutropenia was present in 43/82 episodes. 11 episodes were complicated by bacteremia, predominately with Staphylococcus non- aureus (n = 8). Wound cultures were performed in 36 episodes from 23 patients yielding 99 separate isolates. Gram-negative isolates were most common overall. Modification of antibiotic therapy with further anaerobic coverage was administered successfully in 39/77 episodes. CONCLUSION: This study illustrates that anorectal infections in cancer patients can be successfully managed with antibiotic therapy and local care; surgery can be withheld unless there is evidence of progressive infection or substantial fluctuance and necrosis.

Adolescent↗

Immunoreactivity of Hu proteins facilitates identification of myenteric neurones in guinea-pig small intestine.

Hu proteins, together with neurone-specific enolase (NSE), protein gene product 9.5 (PGP-9.5), microtubule-associated protein-2 (MAP-2) and tubulin beta III isoform, were evaluated immunohistochemically as neuronal markers in whole-mount preparations and cultures obtained from the myenteric plexus of guinea-pig small intestine. Anti-Hu immunostaining marked the ganglion cell somas and nuclei without staining of the neuronal processes in the whole-mounts and cultures. The ganglion cell bodies were not obscured by staining of multiple neuronal fibres and this facilitated accurate counting of the neurones. MAP2 immunostaining also provided clear images of individual neurones in both whole mounts and cultures. Immunoreactivity for NSE, PGP-9.5 and tubulin beta III isoform provided sharp images of the ganglion cells in culture, but not in whole-mount preparations. Strong staining of the neuronal processes in the whole-mount preparations obscured the profiles of the ganglion cell bodies to such an extent that accurate counting of the total neuronal population was compromised. Anti-Hu immunostaining was judged to be an acceptable method for obtaining reliable estimates of total numbers of myenteric neurones in relation to other specific histochemical properties such as histamine binding.

Animals↗

Sensory-motor responses to volume-controlled duodenal distension.

Abstract Visceral perception and secondary peristalsis evoked by distension of the duodenum were studied in 10 healthy volunteers. An impedance planimetric probe for cross-sectional area (CSA) measurements inside a balloon and with three pressure channels was used. Balloon distensions were performed in the fed state with or without the administration of the antimuscarinic drug butylscopolamine. A modified questionnaire was used to assess the nonpainful and painful sensations. The total tension (T(total)) and the passive tension (T(passive)) were determined from the distensions without and with the administration of butylscopolamine, respectively. The active tension (T(active)) was T(total) - T(passive). The stepwise balloon distensions induced the first sensation at a volume of 33 +/- 3 mL. After administration of butylscopolamine the first sensation appeared at 42 +/- 1 mL. The perception score (PS) revealed an approximately linear increase as function of volume, CSA, pressure and tension after the first sensation. Butylscopolamine resulted in significant changes in PS score as function of volume, CSA and strain, but not as a function of pressure and tension. The frequency of the secondary peristalsis increased to the highest value (8.2 +/- 0.8 contractions min(-1)) at a volume of 21 mL. Butylscopolamine almost abolished the distension-evoked motility. T(total) and T(passive) increased nonlinearly as a function of volume, whereas T(active) increased up to a distension volume of 33 mL and then decreased at higher volumes. Hence, the conventional length-tension diagrams as known from studies of smooth muscle strips in vitro can be reproduced in the human duodenum in vivo. This new way of studying intestinal sensation and motility may prove to have both basic and clinical importance as both passive tissue structures and the sensorimotor function are tested.

Adult↗

Mechanical properties in the human gastric antrum using B-mode ultrasonography and antral distension.

The aims of this study were to investigate gastric antral geometry and stress-strain properties by using transabdominal ultrasound scanning during volume-controlled distensions in the human gastric antrum. Seven healthy volunteers underwent stepwise inflation of a bag located in the antrum with volumes up to 60 ml. The stretch ratio and Cauchy stress and strain were calculated from measurements of pressure, diameter, and wall thickness. A second distension series was conducted in three volunteers during administration of the anticholinergic drug butylscopolamine. Analysis of stretch ratios demonstrated positive strain in the circumferential direction, negative strain in the radial direction, and no strain in the longitudinal direction. The stress-strain relation was exponential and did not differ without or with the administration of butylscopolamine. The wall stress was decomposed into its active and passive components. The well-known length-tension diagram from in vitro studies of smooth muscle strips was reproduced. The maximum active tension appeared at a volume of 50 ml, corresponding to a stretch ratio of 1.5. We conclude that the method provides measures of antral biomechanical wall properties and can be used to reproduce the muscle length-tension diagram in humans.

Aged↗

[Genetic polymorphism analysis of apolipoprotein CII microsatellite DNA (TG) n (AG) m in patients with coronary heart disease].

OBJECTIVE: To investigate the relationship between the polymorphism of apolipoprotein CII (apoCII) microsatellite DNA (TG) n (AG) m and coronary heart disease (CHD). METHODS: The apoCII microsatellite DNA (TG) n(AG)m genotypes of 108 patients with CHD and 231 healthy persons were detected by using polymerase chain reaction (PCR) and high voltage denatured polyacrylamid gels electrophoresis. RESULTS: The allele frequency distribution was significantly different between controls and patients with CHD (chi 2 = 19.93, P < 0.05). The allele 17 frequency in CHD group was remarkably higher than that in controls (chi 2 = 13.98, P < 0.01). The concentrations of TG and HDL-C among different genotype groups were significantly different. The levels of TG in 17/29 group and 17/30 group were higher than those in other groups (P < 0.05), while the concentration of HDL-C was lower (P < 0.05). CONCLUSION: The allele 17 of apoCII may be an independent risk factor for CHD in Han population.

Adult↗

P2X(7) receptors in the enteric nervous system of guinea-pig small intestine.

The P2X(7) purinergic receptor subtype has been cloned and emphasized as a prototypic P2Z receptor involved in neurotransmission in the central nervous system and ATP-mediated lysis of macrophages in the immune system. Less is known about the neurobiology of P2X(7) receptors in the enteric nervous system (ENS). We studied the distribution of the receptor with indirect immunofluorescence and used selective agonists and antagonists to analyze pharmacologic aspects of its electrophysiologic behavior as determined with intracellular "sharp" microelectrodes and patch-clamp recording methods in neurons identified morphologically by biocytin injection in the ENS. Application of ATP or 2'- (or-3'-) O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzBzATP) activated an inward current in myenteric neurons. Brilliant blue G, a selective P2X(7) antagonist, suppressed the responses to both agonists. Potency of the antagonist was greatest (smaller IC(50)) for the current evoked by BzBzATP. The P2X(7) antagonists 1-[N,O-bis (1,5-isoquinolinesulfonyl)-N-methyl-l-tyrosyl]-4-piperazine (KN-62) and oxidized ATP also suppressed the BzBzATP-activated current. Micropressure application of BzBzATP evoked rapidly activating depolarizing responses in intracellular studies with "sharp" microelectrodes. Oxidized-ATP suppressed these responses in both myenteric and submucosal neurons. Rapidly activating depolarizing responses evoked by application of nicotinic, serotonergic 5-HT(3), or gamma-aminobutyric acid A (GABA(A)) receptor agonists were unaffected by brilliant blue G. Immunoreactivity for the P2X(7) receptor was widely distributed surrounding ganglion cell bodies and associated with nerve fibers in both myenteric and submucous plexuses. P2X(7) immunoreactivity was colocalized with synapsin and synaptophysin and surrounded ganglion cells that contained either calbindin, calretinin, neuropeptide Y, substance P, or nitric oxide synthase. The mucosa, submucosal blood vessels, and the circular muscle coat also showed P2X(7) receptor immunoreactivity.

Animals↗

Synthesis of solid-supported mirror-image sugars: a novel method for selecting receptors for cellular-surface carbohydrates.

We introduced a novel method, through mirror-image phage display, for the identification of high-affinity D-peptides to target specific cell-surface carbohydrates. Both 3-deoxy-alpha-L-manno-2-octulosonic acid (L-KDO) and L-sialic acid and an L-sialo-disaccharide have been synthesized and attached to a solid support for selection of high-affinity peptide binders displayed on phages. Our initial studies in this effort produce single-chain Fab sequences and dodecapeptides that bind to sialic acid and KDO with nanomolar and high micromolar affinity.

Amino Acid Sequence↗

NMDA receptor activation results in tyrosine phosphorylation of NMDA receptor subunit 2A(NR2A) and interaction of Pyk2 and Src with NR2A after transient cerebral ischemia and reperfusion.

Transient ischemia increases tyrosine phosphorylation of N-methyl-D-aspartate (NMDA) receptor. Several tyrosine kinases are involved in this process. In this study, effect of ischemia and reperfusion (I/R) on tyrosine phosphorylation of NMDA receptor subunit 2A (NR2A) and the interaction of two tyrosine kinases, Src and Pyk2, with NR2A was investigated. Four-vessel occlusion was used to produce transient (15 min) cerebral ischemia in SD rats. Tyrosine phosphorylation of NR2A in hippocampus was enhanced after 15 min of reperfusion and reached its peak level at 6 h of reperfusion. The increase sustained for at least 24 h. Src and Pyk2 co-immunoprecipitated with NR2A and the binding increased after I/R, which also reached a peak at 6 h of reperfusion. Besides, Src and Pyk2 were activated after I/R. These increases were prevented by ketamine, a selective NMDA receptor antagonist, which was administered to the SD rats 20 min before ischemia. Moreover, Src and Pyk2 coprecipitated with each other. These data show that NR2A, Src and Pyk2 might form a protein complex in vivo and the interaction suggests a possible mechanism of signal transduction in the postischemic hippocampus.

Animals↗

CMV-beta-actin promoter directs higher expression from an adeno-associated viral vector in the liver than the cytomegalovirus or elongation factor 1 alpha promoter and results in therapeutic levels of human factor X in mice.

Although AAV vectors show promise for hepatic gene therapy, the optimal transcriptional regulatory elements have not yet been identified. In this study, we show that an AAV vector with the CMV enhancer/chicken beta-actin promoter results in 9.5-fold higher expression after portal vein injection than an AAV vector with the EF1 alpha promoter, and 137-fold higher expression than an AAV vector with the CMV promoter/enhancer. Although induction of the acute-phase response with the administration of lipopolysaccharide (LPS) activated the CMV promoter/enhancer from the context of an adenoviral vector in a previous study, LPS resulted in only a modest induction of this promoter from an AAV vector in vivo. An AAV vector with the CMV-beta-actin promoter upstream of the coagulation protein human factor X (hFX) was injected intravenously into neonatal mice. This resulted in expression of hFX at 548 ng/ml (6.8% of normal) for up to 1.2 years, and 0.6 copies of AAV vector per diploid genome in the liver at the time of sacrifice. Neonatal intramuscular injection resulted in expression of hFX at 248 ng/ml (3.1% of normal), which derived from both liver and muscle. We conclude that neonatal gene therapy with an AAV vector with the CMV-beta-actin promoter might correct hemophilia due to hFX deficiency.

Actins↗

Cellular interactions in the development of the olfactory system: an ablation and homotypic transplantation analysis.

In the current study, we addressed two questions: First, is the olfactory placode necessary for the development of the olfactory bulb and the entire telencephalon? Second, does the olfactory placode contribute cells to the olfactory bulb? We addressed these questions by unilaterally ablating the olfactory placode in chick embryos before an olfactory nerve was produced and, in a second series of experiments, by replacing the ablated chick olfactory placode with a quail olfactory placode. Our results indicate that the olfactory placode is critical for olfactory bulb development, but is not necessary for the development of the rest of the telencephalon. Further, our results support the hypothesis that LHRH neurons and olfactory nerve glia originate in the olfactory placode, but do not support an olfactory placodal origin for other cell types within the olfactory bulb.

Animals↗

Lipopolysaccharide potentiates the effect of hepatocyte growth factor upon replication in lung, thyroid, spleen, and colon in rats in vivo.

Induction of replication may potentiate in vivo gene therapy, as some viral vectors only transduce dividing cells. Hepatocyte growth factor (HGF) increases the percentage of replicating hepatocytes to 18-fold that in normal rats, and lipopolysaccharide (LPS) modestly potentiates this effect. In this study, the effect of iv HGF upon replication in other organs was determined. HGF at 10 mg/kg resulted in replication that was < or =3-fold that of normal rats in alveolar and proximal renal tubular cells. HGF alone had no effect upon replication of epithelial cells from the bronchi, thyroid, pancreas, or colon or upon cells from the muscle, pancreatic islets, spleen, blood vessels, or thymus. HGF and LPS at 5 mg/kg resulted in replication that was 9-fold that of normal rats in alveolar cells, 25-fold in bronchial epithelial cells, 4-fold in thyroid epithelial cells, 1.5-fold in the red pulp of the spleen, and 2-fold in colonic epithelial cells. The synergistic effect may be due to the fact that LPS upregulated the HGF receptor c-met in thyroid, spleen, and colon. We conclude that iv administration of HGF alone is relatively specific for inducing hepatocyte replication and would allow selective gene transfer into the liver.

Animals↗

ISMh2, a novel insertion sequence-like element associated with nifA from Mesorhizobium huakuii.

Sequence analysis of the nifA gene, including its 5' flanking region, of Mesorhizobium huakuii revealed the presence of a novel IS-like element called ISMh2. It is 828 bp in length and possesses two imperfect terminal inverted repeats of 14 bp with only one mismatch. The putative transposase encoded by ISMh2 is composed of 204 amino acids. In comparison with other insertion sequences ISMh2 likely belongs to the IS6 family. Multiple copies of ISMh2 were detected in the genome of M. huakuii 159 by Southern hybridization. RT-PCR analysis showed that nifA and ISMh2 cotranscribed. Attempts to detect the transposition ability of ISMh2 were unsuccessful.

Alphaproteobacteria↗

Establishment of a human hepatocyte line (OUMS-29) having CYP 1A1 and 1A2 activities from fetal liver tissue by transfection of SV40 LT.

Immortalized human hepatocytes that can retain functions of drug-metabolizing enzymes would be useful for medical and pharmacological studies and for constructing an artificial liver. The aim of this study was to establish immortalized human hepatocyte lines having differentiated liver-specific functions. pSVneo deoxyribonucleic acid, which contains large and small T genes in the early region of simian virus 40, was introduced into hepatocytes that had been obtained from the liver of a 21-wk-old fetus. Neomycin-resistant immortalized colonies were cloned and expanded to mass cultures to examine hepatic functions. Cells were cultured in a chemically defined serum-free medium, ASF104, which contains no peptides other than recombinant human transferrin and insulin. As a result, an immortal human hepatocyte cell line (OUMS-29) having liver-specific functions was established from one of the 13 clones. Expression of CYP 1A1 and 1A2 messenger ribonucleic acid by the cells was induced by treatment with benz[a]pyrene, 3-methylcholanthrene, and benz[a]anthracene. OUMS-29 cells had both the polycyclic aromatic hydrocarbon receptor (AhR) and AhR nuclear translocator. Consequently 7-ethoxyresorufin deethylase activity of the cells was induced time- and dose-dependently by these polycyclic aromatic hydrocarbons. This cell line is expected to be instrumental as an alternative method in animal experiments for studying hepatocarcinogenesis, drug metabolisms of liver cells, and hepatic toxicology.

Cell Line↗

Sialogogic activity in the rat of peptides analogous to [Tyr8]-substance P in which substitutions have been made in the N-terminal amino acids.

In order to elucidate the regulatory roles for salivation of amino acids in positions 1-4 of the N-terminal region of [Tyr8]-substance P (SP), the structure-sialogogic activity correlations of various synthetic octa- to undecapeptides replaced in positions 1-4 of [Tyr8]-SP with each of 19 common amino acids, one by one, and with the same sequence of the C-terminal hepatapeptide as that of [Tyr8]-SP, were studied in the submandibular glands of rats after intraperitoneal injection. Each of 19 octa-, nona-, deca- and undecapeptides with replaced amino acids and a penta- to decapeptide with the progressive elimination of the N-terminal portion were newly synthesized by the multipin peptide method. All octa- to undecapeptides replaced with each of 19 common amino acids in positions 1-4 had sialogogic activities. In 19 octa- and decapeptides in which P4 and P2 had been replaced, four and three replacements, respectively, had significantly increased secretory activities. In contrast, in 19 nonapeptides in which K3 had been replaced, none had significantly increased secretory activities. Furthermore, in 19 undecapeptides in which R1 had been replaced, most replacements had significantly increased or equipotent activities for fluid secretion. It is concluded that amino acids in the N-terminal region of various tachykinins may not need to be strictly conserved and that amino acid residues in the N-terminal portion, R1 in particular and P2, may strongly inhibit secretory activity.

Amino Acid Substitution↗

Expression and postnatal changes of adrenergic receptor subtype mRNA in rat submandibular glands.

Adrenergic receptors (ARs) are involved in regulating saliva secretion and composition in salivary glands. Nine AR subtypes, including three alpha1-ARs (alpha1a-, alpha1b- and alpha1d-ARs), three alpha2-ARs (alpha2A-, alpha2B- and alpha2C-ARs) and three beta-ARs (beta1,beta2- and beta3-ARs), have been identified through molecular cloning. The five subtype genes, alpha1a-, alpha1b-, alpha2A-, beta1-, and beta2-ARs, were expressed in rat submandibular glands. In contrast, the other four subtype mRNAs, alpha1d-, alpha2B-, alpha2C- and beta3-ARs, were not detected by reverse transcription-polymerase chain reaction (RT-PCR). The steady-state mRNA expression for the five AR subtypes in rat submandibular glands was measured by quantitative competitive RT-PCR using synthetic DNA as internal standard at different stages of postnatal development. The relative rank order of AR subtype mRNA expression was alpha1a>beta2>beta1>alpha2A>alpha1b at all stages except that beta1- and alpha2A-subtypes were reversed at 2 weeks of age. The gene expression of alpha1a-AR subtype relative to total AR was low at 2 weeks of age and increased and reached a maximum at 6 weeks of age, whereas those patterns of alpha2A-, beta1- and beta2-AR subtypes were similar to each other and their gene expressions were high at 2 weeks of age and then decreased. On the other hand, the gene expression of alpha1b-AR subtype did not change over the different stages in relation to that of a housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase, and to total AR. Although rat submandibular glands contain the five AR subtype mRNAs, distinct subtype-specific expression is evident.

Age Factors↗

The extracellular matrix protein lacunin is expressed by a subset of hemocytes involved in basal lamina morphogenesis.

The extracellular matrix protein of Manduca sexta known as lacunin is localized to basal laminae and granular cells of the hemolymph. Circulating granular cells increase in both number and size as pupal basal laminae break down after the initiation of adult development. Basal laminae of wing epithelia break down as ecdysteroid levels rise during adult development, and lacunin immunoreactivity concurrently passes from basal laminae to endocytic vacuoles of granular cells. Granular cells not only endocytose lacunin protein that they salvage from the remnants of the old basal laminae but they also express transcripts for lacunin. As new adult basal laminae form several days later, the number of circulating granular cells decreases as lacunin immunoreactivity appears in the new basal laminae.

Journal Article↗