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Biomedical subjects

X M Wang

Publications and source records attributed to X M Wang.

At least 55 records · Page 3Linked to original sources

Endomorphin-1 and endomorphin-2 modulate responses of trigeminal neurons evoked by N-methyl-D-aspartic acid and somatosensory stimuli.

The present study investigated the modulation of N-methyl-D-aspartate (NMDA)-evoked and peripheral cutaneous stimulus-evoked responses of trigeminal neurons by endomorphins, endogenous ligands for the mu-opioid receptor. Effects of endomorphins, administered microiontophoretically, were tested on the responses of nociceptive neurons recorded in the superficial and deeper dorsal horn of the medulla (trigeminal nucleus caudalis) in anesthetized rats. Endomorphin-1 and endomorphin-2 predominantly reduced the NMDA-evoked responses, producing an inhibitory effect of 54.1 +/- 2.96% (mean +/- SE; n = 34, P < 0.001) in 92% (34/37) of neurons and 63.6 +/- 3.61% (n = 32, P < 0.001) in 91% (32/35) of neurons, respectively. The inhibitory effect of endomorphins was modality specific; noxious stimulus-evoked responses were reduced more than nonnoxious stimulus-evoked responses. Naloxone applied at iontophoretic current that blocked the inhibitory effect of [D-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin, reduced the peak inhibitory effect of endomorphins on the NMDA- and natural stimulus-evoked responses. We suggest that endomorphins by acting at micro-opioid receptor selectively modulate noxious stimulus-evoked responses in the medullary dorsal horn.

Analgesics, Opioid↗

Correlation of Glut-1 glucose transporter expression with.

Positron emission tomography (PET) with [18F]2-fluoro-2-deoxy-D-glucose (FDG) may show negative results for bronchioloalveolar lung carcinoma. We investigated the correlation of Glut-1 glucose transporter expression with [18F]FDG uptake in non-small cell lung cancer. Thirty-two patients with 34 non-small cell lung cancers (7 bronchioloalveolar carcinomas, 23 non-bronchioloalveolar adenocarcinomas, 3 squamous cell carcinomas, and 1 adenosquamous cell carcinoma) were studied. Final diagnoses were established by histology (via thoracotomy) in all patients. [18F]FDG PET was performed 40 min after i.v. injection of 185 MBq [18F]FDG. For semi-quantitative analysis of [18F]FDG uptake, standardized uptake values (SUVs) were calculated. Glut-1 expression was studied in terms of the immunohistochemistry of paraffin sections using anti-Glut-1 antibody to determine the intensity (0-3) of Glut-1 immunoreactivity and percentage of the Glut-1-positive area. Of seven bronchioloalveolar carcinomas, six (85.7%) were negative for the expression of Glut-1, while only one (4.3%) of 23 non-bronchioloalveolar adenocarcinomas was negative (P < 0.0001). The percentages of Glut-1-positive area, as well as the SUVs, were significantly lower in bronchioloalveolar carcinomas (n = 7) (2.86% +/- 7.56% and 1.25 +/- 0.75, respectively) than in non-bronchioloalveolar adenocarcinomas (n = 23) (54.83% +/- 25.64%, P < 0.0001, and 3.94 +/- 1.93, P = 0.001, respectively). The degree of cell differentiation correlated with the percentage of Glut-1-positive area and SUVs in adenocarcinoma of the lung. Correlations between SUVs and the intensity of Glut-1 immunoreactivity were also significant (intensities 0 and 1, n = 11, SUV 1.47 +/- 0.63; intensities 2 and 3, n=23, SUV 4.78 +/- 2.13; P < 0.0001). The percentage of Glut-1-positive area correlated significantly with SUVs (n = 34, r = 0.658, P < 0.01). Overexpression of Glut-1 correlated with high [18F]FDG uptake. These findings suggest that Glut-1 expression is related to [18F]FDG uptake in non-small cell lung cancer. Glut-1 expression, as well as [18F]FDG uptake, correlated with the degree of cell differentiation in adenocarcinomas, and both Glut-1 expression and [18F]FDG uptake were significantly lower in bronchioloalveolar carcinomas than in non-bronchioloalveolar carcinomas.

Adenocarcinoma, Bronchiolo-Alveolar↗

[Expression of 3'-truncated preS/S gene in transgenic mice].

The 3'-truncated preS/S region from HBV genome, encoding a transcriptional transactivator, was cloned and a recombination expression vector for 3'-truncated HBV preS/S sequences under the control of the CMV promoter was constructed by recombination DNA techniques. Then, the expression vector DNA was microinjected into pronuclei of fertilized mouse oocytes. Founders of transgenic mice harbouring the recombination gene which can be expressed were screened by polymerase chain reaction (PCR) at genomic DNA level and confirmed by ELISA andlysis at protein level. Two of 15 mice in one series of microinjections showed the expression of 3'-truncated preS/S gene from HBV genome. The expression vector of 3'-truncated preS/S gene might be helpful for further studies of relationship between the expression production of 3'-truncated preS/S sequence and HBV-associated oncogenesis in vitro. The transgenic mice with expressing 3'-truncated preS/S gene will provide new insight into its roles in the development of human hepatocelluar carcinoma (HCC) in vivo.

Animals↗

Apoptosis of osteoclast-like cells induced by alendronate is related to Fas gene expression.

OBJECTIVE: To detect the mechanism of alendronate during inhibition of bone resorption by inducing apoptosis of osteoclasts and its relationship with the Fas gene. METHODS: The osteoclast-like cells (OCLs) from a giant cell tumor in vitro were used. After treatment with alendronate for 48 hours, the OCLs were identified by fluorescence microscope, transmission electron microscope, in situ end labeling (TUNEL), HE staining, in situ hybridization with Dig-Fas Probe, and immunohistochemistry with anti-Fas antibodies. RESULTS: The detachment rate of OCLs was 76% after exposure to alendronate, while the control cell detachment rate was only 3%. The nonadherent OCLs showed the features of cell contraction, pyknosis, chromatin condensation connecting with nuclear membrane to form circle or crescent bodies, and occasional nuclear fragmentation. The detached OCLs were positive by in situ end labeling, and the control group was negative. Nonadherent OCLs were ultrastructurally consistent with apoptosis (shrunken cells with pyknotic nuclei, chromatin condensation connecting with nuclear membrane, mitochondria recruitment with intact structure, and loosening of endoplasmic reticulum). The results by in situ hybridization and immunohistochemistry indicated that the Fas gene expression of nonadherent OCLs were positive, but the controls were negative. CONCLUSION: Alendronate promotes apoptosis of the OCLs and it is related with the expression of the Fas gene.

Alendronate↗

[Effect of acetylcholine on the discharge of presympathetic neurons of the rostral ventrolateral medulla of rats].

Effect of acetylcholine (ACh) on presympathetic neurons of the rostral ventrolateral medulla (RVLM) was studied by extracellular recording technique and microiontophoresis in urethane-anaesthetized rats. Spontaneous discharges of 35 presympathetic neurons of RVLM were recorded. All the neurons showed increased discharge by microiontophoresis of ACh dose- dependently with the increase of iontophoretic current. In 22 units microiontophoresis of atropine decreased the baseline discharge rate and completely blocked the excitatory effect of ACh on these neurons, while in the remaining 7 and 6 neurons microiontophoresis of d-tubocurarine (d-TC) and hexamethonium (C(6)) respectively had no effect and did not block the action of ACh. The above finding suggests that presympathetic neurons of RVLM receive a tonic excitatory cholinergic input mediated by M receptors.

Acetylcholine↗

Rescue of axotomized rubrospinal neurons by brain-derived neurotrophic factor (BDNF) in the developing opossum, Didelphis virginiana.

Many rubrospinal neurons die in developing opossums when their axon is cut at thoracic levels of the spinal cord and in the present study we asked whether they can be rescued by brain-derived neurotrophic factor (BDNF). Bilateral injections of Fast Blue (FB) were made into the rostral lumbar cord to prelabel rubrospinal neurons and 5 days later the rubrospinal tract was cut unilaterally by hemisecting the thoracic cord. Immediately after hemisection, BDNF-soaked gelfoam was placed into the lesion cavity. Since pilot data indicated that one application of BDNF was not sufficient to produce a rescue effect, a second application was made 7 days later. Seven days after the second application the pups were killed by an overdose of anesthetic so that the red nucleus contralateral and ipsilateral to the lesion site could be examined for labeled neurons. The rubrospinal tract is almost entirely crossed, so the red nucleus contralateral to the lesion contained many axotomized neurons, whereas the red nucleus ipsilateral to it did not. Age-matched controls were subjected to the same procedures, but the gelfoam applied to the lesion site in the experimental animals was soaked only in the vehicle used to deliver BDNF. In all cases, labeled neurons were fewer in number in the red nucleus contralateral to the lesion than ipsilateral to it. It was of particular interest, however, that labeled neurons contralateral to the lesion were more numerous in the animals treated with BDNF than in the controls. We conclude that BDNF rescues at least some rubrospinal neurons from axotomy-induced cell death in developing opossums suggesting that loss of access to BDNF, and perhaps other neurotrophins, contributes to failure of rubrospinal neurons to survive axotomy.

Amidines↗

Acute intermittent morphine increases preprodynorphin and kappa opioid receptor mRNA levels in the rat brain.

We determined the effects of morphine on mRNA levels for the opioid ligands preprodynorphin (PPD) and preproenkephalin (PPE) and the kappa opioid receptor (KOR). Rats received six injections of morphine (6.25 mg/kg/injection) every 2 h, and were sacrificed 30 min later. mRNA levels were measured in brain tissue after removal of the cortex, cerebellum and brainstem. There were increases in PPD and KOR mRNA levels (P<0.05 and P<0.005, respectively), with no alteration of PPE. These alterations in the kappa/dynorphin system may counter morphine-induced effects on the brain.

Animals↗

Developmental plasticity of ascending spinal axons studies using the North American opossum, Didelphis virginiana.

The objectives of the present study were to determine if axons of all ascending tracts grow through the lesion after transection of the thoracic spinal cord during development in the North American opossum, and if so, whether they reach regions of the brain they normally innervate. Opossum pups were subjected to transection of the mid-thoracic cord at PD5, PD8, PD12, PD20, or PD26 and injections of Fast Blue (FB) into the lower thoracic or upper lumbar cord 30-40 days or 6 months later. In the PD5 transected cases, labeled axons were present in all of the supraspinal areas labeled by comparable injections in unlesioned, age-matched controls. In the experimental cases, however, labeled axons appeared to be fewer in number and in some areas more restricted in location than in the controls. When lesions were made at PD8, labeled axons were present in the brain of animals allowed to survive 30-40 days prior to FB injections but they were not observed in those allowed to survive 6 months. When lesions were made at PD12 or later, labeled axons were never found rostral to the lesion. It appears, therefore, that axons of all ascending spinal pathways grow though the lesion after transection of the thoracic cord in developing opossums and that they innervate appropriate areas of the brain. Interestingly, the critical period for such growth is shorter than that for most descending axons, suggesting that factors which influence loss of developmental plasticity are not the same for all axons.

Afferent Pathways↗

Laboratory diagnosis of toxigenic Clostridium difficile by polymerase chain reaction: presence of toxin genes and their stable expression in toxigenic isolates from Japanese individuals.

Clostridium difficile causes pseudomembranous colitis and antibiotic-associated diarrhea. The definitive diagnosis of C. difficile infection is finally accomplished by the isolation of toxigenic C. difficile. However, only a small number of Japanese clinical laboratories are able to reach a definitive diagnosis of C. difficile infection, probably because simple reliable assays for toxins in the isolates are not available. In this study, we examined the compatibility of a polymerase chain reaction (PCR) assay and tissue culture assay to identify toxigenic C. difficile, in toxigenic and nontoxigenic C. difficile isolates from Japanese patients and healthy carriers. The specificity of PCR primers was demonstrated by restriction endonuclease digestion and seminested PCR in C. difficile VPI 10463 strain. No PCR product was amplified in the eight other clostridial species used to check the specificity of the PCR assay. The detection limit was 10(3) cells. Both toxin A and toxin B genes (the genes encoding the major virulence factors of C. difficile) were detected in 58 toxigenic C. difficile isolates, which showed a wide range of cytotoxic activity in tissue culture assays. Neither of the toxin genes was carried by 40 nontoxigenic strains of C. difficile. The results of this study strongly suggest that a definitive diagnosis of C. difficile infection can be accomplished by PCR detection of the toxin genes rather than by tissue culture assay of isolates.

Adult↗

Orphanin FQ (nociceptin) modulates responses of trigeminal neurons evoked by excitatory amino acids and somatosensory stimuli, and blocks the substance P-induced facilitation of N-methyl-D-aspartate-evoked responses.

The present investigation details the modulation of medullary dorsal horn neuron responses to excitatory amino acids and peripheral cutaneous stimuli by orphanin FQ (nociceptin), an endogenous ligand for the opioid receptor-like, receptor. Effects of orphanin FQ, administered microiontophoretically or given intracerebroventricularly, were tested on the responses of nociceptive-specific, wide dynamic range and low threshold neurons recorded in the superficial and deeper dorsal horn of the medulla (trigeminal nucleus caudalis) in anesthetized (urethane or pentobarbital) male rats. Microiontophoretic application of orphanin FQ reduced the N-methyl-D-aspartate-evoked responses in 86% (71/82) of neurons, and the (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid-evoked responses in 86% (30/35) of neurons. However, orphanin FQ produced a longer lasting inhibitory effect on the N-methyl-D-aspartate-evoked responses relative to the (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid-evoked responses. The inhibitory effect of orphanin FQ was not modality-specific, responses evoked by noxious as well as non-noxious stimuli were reduced in 22/23 neurons. However, the inhibitory effect was more pronounced on noxious stimulus-evoked responses. Naloxone applied at currents that antagonized the inhibitory effects of selective agonists at mu and kappa opioid receptors failed to inhibit the effects of orphanin FQ. Microiontophoretic co-application of substance P with N-methyl-D-aspartate facilitated the N-methyl-D-aspartate-evoked responses in 52% (26/50) of nociceptive neurons. Orphanin FQ blocked or reduced the substance P-induced facilitation by 86+/-24.4% (n = 14). In order to compare electrophysiological data with previous behavioral observations, effects of orphanin FQ administered intracerebroventricularly were tested on the excitatory amino acid-evoked responses. Orphanin FQ reduced the N-methyl-D-aspartate-evoked responses in 85% (11/13) of neurons whereas the (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid-evoked responses were facilitated in 69% (9/13) of neurons. We suggest that orphanin FQ produces a predominantly inhibitory effect on, (i) noxious stimuli evoked responses, (ii) excitatory amino acid receptor-mediated transmission and, (iii) the substance P-induced facilitation of the N-methyl-D-aspartate-evoked responses. We conclude that orphanin FQ primarily produced an antinociceptive action at the level of the dorsal horn of the medulla.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

Characterization of early developmental pattern of expression of neurotensin/neuromedin N gene in foregut and midgut.

The gut endocrine gene, neurotensin (NT/N), is expressed in a strict temporally and spatially specific pattern. Utilizing a sensitive reverse transcription-polymerase chain reaction procedure, we analyzed foregut and midgut organs for NT/N expression, determined the earliest apparent time point that NT/N is expressed in the gastrointestinal tract and compared the temporal relationship of NT/N with other genes. NT/N expression was detected in the fetal and early postnatal stomach. In the small bowel, NT/N was expressed at the earliest fetal time (14 days) that the small bowel could be reliably delineated from other gut organs; in contrast, expression of sucrase-isomaltase was only apparent at 28 days after birth. NT/N was expressed in the fetal (12, 14 and 16 days) liver and then again on days 3 and 7 after birth; however, NT/N was detected only in the fetal (14 and 16 days) pancreas. Finally, NT/N expression was first detected at approximately 12 days gestation in the primitive foregut and midgut, thus occurring significantly earlier than actual intestinal cytodifferentiation. NT/N is widely expressed in the gastrointestinal tract during fetal development, suggesting the presence of a shared ancestral stem cell; NT/N expression is then restricted to the small bowel of the adult. The determination of the cellular factors regulating the expression of NT/N will provide a better understanding of the mechanisms responsible for the strict patterning of gene expression in the gastrointestinal tract and possibly gut differentiation.

Animals↗

The selective cytotoxicity of cobra venom factor immunoconjugate on cultured human nasopharyngeal carcinoma cell line.

The selective cytotoxicity of a CVF immunoconjugate on human nasopharyngeal carcinoma cell line was reported. Cobra venom factor (CVF), a C3b-like glycoprotein, was linked to BAC5, a murine monoclonal antibody directed against a human nasopharyngeal carcinoma-associated membrane antigen, by a disulfide bond. The high affinity to cultured human nasopharyngeal cells (CNE2) and the complement activating potency retained in CVF immunoconjugate. Although the equimolar concentration of BAC5 or CVF alone was harmless to CNE2 cells, the CVF immunoconjugate in the presence of fresh human serum exhibited selective cytotoxicity on CNE2 cells in a concentration- (IC50 3.07 x 10(-7) mol/L) and time-dependent manner. No cytotoxicity occurred on either CNE1 (another human nasopharyngeal carcinoma cell line) or MGC-803 (human gastric carcinoma cell line) cells. Furthermore, direct lytic factor (DLF, cardiotoxin) separated from cobra venom, augmented CVF immunoconjugate-induced cytotoxicity significantly. These results indicate that the CVF immunoconjugate has complement-mediated selective cytotoxicity on CNE2 cells, which can be potentiated by DLF.

Animals↗

Hypothalamic-pituitary-adrenal activity and pro-opiomelanocortin mRNA levels in the hypothalamus and pituitary of the rat are differentially modulated by acute intermittent morphine with or without water restriction stress.

Acute administration of morphine stimulates the secretion of hypothalamic-pituitary-adrenal (HPA) hormones, ACTH, beta-endorphin and corticosterone in the rat. In this study we investigated the effects of repeated multiple-dose morphine on HPA activity under two different conditions: without or with water restriction stress. Rats received six intermittent injections of morphine (6.25 mg/kg per injection, s.c.) every 2 h and were killed 30 min after the last injection. The results were as follows. (1) Morphine significantly elevated plasma ACTH and corticosterone levels; water restriction also significantly increased ACTH secretion, but with no significant increase of plasma corticosterone levels. In contrast, rats treated with morphine under the water restriction condition failed to show any increases of either ACTH or corticosterone levels. (2) Morphine did not change pro-opiomelanocortin (POMC) mRNA levels in the anterior pituitary; whereas water restriction significantly increased the POMC mRNA levels. The water restriction-induced increases of POMC mRNA in the anterior pituitary were absent in the rats which received morphine. (3) Morphine significantly increased POMC mRNA levels in the hypothalamus; water restriction had no effect. The morphine-induced increases in POMC mRNA in the hypothalamus were absent in the rat under the water restriction condition. These findings, that the effects of morphine on HPA activation or POMC mRNA expression depend on the presence of stress, suggest a counter-regulatory role of opiates on a stress response and opioid gene expression.

Adrenocorticotropic Hormone↗

Pharmacokinetics of atenolol enantiomers in 12 Chinese healthy men.

AIM: To study the pharmacokinetics of atenolol (Ate) stereoisomers in Chinese. METHOD: A single oral dose of 100 mg of racemic Ate tablets were given to 12 healthy volunteers of Han nationality. Plasma and urine concentrations were determined by the reversed phase HPLC method. RESULTS: The disposition of d-Ate and l-Ate was conformed to one-compartment model. Maximal plasma concentration (Cmax): l-Ate (331 +/- 79) micrograms.L-1, d-Ate (342 +/- 78) micrograms.L-1. Area under blood concentration-time curve (AUC): d-Ate (2635 +/- 610) micrograms.h.L-1, l-Ate (2442 +/- 588) micrograms.h.L-1. Renal clearance (Clr): l-Ate (6.9 +/- 1.2) L.h-1, d-Ate (6.5 +/- 1.3) L.h-1. CONCLUSION: The disposition of Ate stereoisomers is of stereoselectivity.

Adrenergic beta-Antagonists↗

[A method for preparation of carbon fiber electrode].

Carbon fiber electrode (CFE) can be used in detection of exocytosis of single neuron or endocrine cell. A simplified method for preparation of CFE, as described in the present paper, greatly ensures the coordination and success of the preparation. Such CFEs have a low noise level. Exocytosis of rat adrenal chromaffin cells was investigated in the clarification of the effect of MPP+ on transmitter release. It is found that exocytosis was not stimulated by MPP+ and the high K+ induced secretion was not changed by MPP+. These observations suggest that the previously reported elevation of dopamine content in the surrounding histosolution of DA neuron is probably resulted from blockage of the re-uptake of DA by MPP+.

1-Methyl-4-phenylpyridinium↗