PubMed Health⌕ Search

Biomedical subjects

X H Chen

Publications and source records attributed to X H Chen.

At least 55 records · Page 3Linked to original sources
PubMed1998

CCK(B) receptors in the periaqueductal grey are involved in electroacupuncture antinociception in the rat cold water tail-flick test.

Cholecystokinin octapeptide (CCK-8) (0.25-2.0 ng), the CCK(A) receptor antagonist L-364,718 (60-100 ng) or the CCK(B) receptor antagonist L-365,260 (0.3125-1.25 ng) was administered into the periaqueductal grey (PAG) of male SD rats. The antinociceptive effect induced by electroacupuncture (EA) stimulation of different frequencies was then measured by the cold water tail-flick (CWT) test. The results showed that (1) microinjection of CCK-8 into the PAG can significantly block the antinociceptive effect induced by all frequencies of EA stimulation. The effectiveness of the blockade was 100 > 2 Hz. In addition, CCK-8 blocks the antinociception seen following termination of the electrical stimulation at 100 Hz; (2) microinjection of L-365,260 (1.25 ng) into the PAG significantly increased the 100 Hz EA antinociceptive effect but not the 2 Hz EA antinociceptive effect and microinjection of L-364,718 into PAG did not affect either 2 or 100 Hz EA antinociception. These results demonstrate that CCK-8 in the PAG can antagonize the antinociceptive effect induced by EA stimulation, and the CCK effect is likely to be mediated by the CCK(B) receptor, but not the CCK(A) receptor.

Animals↗
PubMed1998

Magnetic resonance spectroscopy of diffuse brain trauma in the pig.

The acute metabolic events linked to the evolution of selective axonal pathology in the white matter following diffuse brain injury have not previously been evaluated due to the paucity of relevant experimental models. Here, we utilized a new model of inertial brain injury in the pig that selectively damages axons in the white matter, and applied proton and phosphorous magnetic resonance spectroscopy (MRS) to noninvasively monitor the temporal course of metabolic changes following trauma. Evaluating four pigs with MRS prior to injury, within 1 h and 3 and 7 days postinjury, we found that widespread axonal injury was produced in the absence of changes in pH, PCr/Pi, or the concentrations of ATP, and lactate. However, we did observe an acute 60% loss of intracellular Mg2+ levels, which gradually resolved by 7 days postinjury. In addition, we found that the levels of the neuron marker, N-acetylaspartate (NAA), acutely dropped 20% and remained persistently decreased for at least 7 days postinjury. Moreover, the changes in Mg2+ and NAA were found with MRS in the absence of abnormalities with conventional magnetic resonance imaging (MRI). These results show that (1) profound alterations in intracellular metabolism occur acutely following diffuse axonal pathology in the white matter, but in the absence of indicators of ischemia, and (2) axonal pathology may be evaluated with high sensitivity utilizing noninvasive MRS techniques.

Adenosine Triphosphate↗
PubMed1998

Effects of nitroquine on ultrastructures and cytochrome oxidase of exoerythrocytic Plasmodium yoelii in rat liver.

AIM: To study the effects of nitroquine acetate (NA) on the ultrastructures and cytochrome-c oxidase (CCO) of exoerythrocytic forms (EEF) of Plasmodium yoelii. METHODS: Rats were inoculated with sporozoites directly into the liver. After 48 h rats were killed. Rat liver thin sections were incubated in histochemical reaction medium, then examined by transmission electron microscopy. NA (2 mg.kg-1) was fed to rats 3.5 h and 14 h before killing the rats. RESULTS: At 3.5 h, in the parasites there appeared swelling and proliferation of mitochondria, dilation of endoplasmic reticulum, and reduction of the electron density of parasites' nuclei. The structures of the parasites disintegrated to form many autophagocytes 14 h after exposure to NA. The reaction products of CCO still existed until 14 h after using NA. CONCLUSION: CCO was not the starting point of NA action. NA interferes with the structure and function of the cytoplasm and nucleus of malaria parasites and exerts its antimalarial effects in many aspects.

Animals↗
PubMed1997

Aspartate substitutions establish the concerted action of P-region glutamates in repeats I and III in forming the protonation site of L-type Ca2+ channels.

Hydrogen ions reduce ion flux through voltage-gated Ca2+ channels by binding to a single protonation site with an unusually high pKa. Recent evidence localizes the protonation site to the same locus that supports high affinity Ca2+ binding and selectivity, a set of four conserved glutamate residues near the external mouth of the pore. Remaining controversy concerns the question of whether the protonation site arises from a single glutamate, Glu-1086 (EIII), or a combination of Glu-1086 and Glu-334 (EI) working in concert. We tested these hypotheses with individual Glu --> Asp substitutions. The Glu --> Asp replacements in repeats I and III stood out in two ways. First, in both EID and EIIID, protonation was destabilized relative to wild type, whereas it was unchanged in EIID and stabilized in EIVD. The changes in affinity were entirely due to alterations in H+ off-rate. Second, the ratio of protonated conductance to deprotonated conductance was significantly closer to unity for EID and EIIID than for wild-type channels or other Asp mutants. Both results support the idea that EI and EIII act together to stabilize a single titratable H+ ion and behave nearly symmetrically in influencing pore conductance. Neutralization of EIII by alanine replacement clearly failed to abolish susceptibility to protonation, indicating that no single glutamate was absolutely required. Taken together, all the evidence supports a model in which multiple carboxylates work in concert to form a single high affinity protonation site.

Animals↗
PubMed1997

Jak1 expression is required for mediating interleukin-4-induced tyrosine phosphorylation of insulin receptor substrate and Stat6 signaling molecules.

The Jak1, Jak2, Jak3, and Fes tyrosine kinases have been demonstrated to undergo tyrosine phosphorylation in response to interleukin (IL)-4 stimulation in different cell systems. However, it is not clear which, if any, of these kinases are responsible for initiating IL-4-induced tyrosine phosphorylation of intracellular substrates in vivo. In the present study, we have utilized a mutant Jak1-deficient HeLa cell line, E1C3, and its parental Jak1-expressing counterpart, 1D4, to analyze the role of Jak1 in mediating IL-4-induced tyrosine phosphorylation events. IL-4 treatment rapidly induced tyrosine phosphorylation of insulin receptor substrate (IRS)-1 and IRS-2 in 1D4 but not in E1C3 cells. IL-4-mediated tyrosine phosphorylation of Stat6 was pronounced in 1D4 cells, while no IL-4-induced Stat6 phosphorylation was detected in E1C3 cells. IL-4 also induced Stat6 DNA binding activity from lysates of 1D4 but not E1C3 cells utilizing a radiolabeled immunoglobulin heavy chain germline epsilon promotor sequence (Iepsilon) in an electrophoretic mobility shift assay. Reconstitution of Jak1 expression in E1C3 cells restored the ability of IL-4 to induce IRS and Stat6 tyrosine phosphorylation. These results provide evidence that Jak1 expression is required for mediating tyrosine phosphorylation and activation of crucial molecules involved in IL-4 signal transduction.

HeLa Cells↗
PubMed1997

Progressive atrophy and neuron death for one year following brain trauma in the rat.

Although atrophic changes have been well described following traumatic brain injury (TBI) in humans, little is known concerning the mechanisms or progression of brain tissue loss. In the present study, we evaluated the temporal profile of histopathological changes following parasagittal fluid-percussion (FP) brain injury in rats over 1 year postinjury. Anesthetized 3-4 month-old Sprague-Dawley Rats (n = 51) were subjected to FP brain injury of high severity (2.5-2.9 atm, n = 51) or sham treatment (n = 27). At 1 h, 2 h, 48 h, 1 week, 2 weeks, 1 month, 2 months, 6 months and 1 year after brain injury or sham treatment, these animals were humanely euthanized. Brain sections were analyzed with image-processing techniques to determine the extent of cortical tissue loss and shrinkage of the hippocampal pyramidal cell layer. In addition, cell counting was performed to determine the number of neurons in the dentate hilus of the hippocampus, and glial fibrillary acidic protein (GFAP) immunostaining was used to reveal reactive astrocytosis. Examination of the injured brains revealed substantial and progressive tissue loss with concomitant ventriculomegaly in the hemisphere ipsilateral to injury. The regions with the most notable progressive atrophy included the cortex, hippocampus, thalamus, and septum. Quantitative analysis demonstrated a significantly progressive loss of cortical tissue as well as shrinkage of the hippocampal pyramidal cell layer ipsilateral to injury over 1 year following injury. In addition, reactive astrocytosis in regions of atrophy and progressive bilateral death of neurons in the dentate hilus was observed for 1 year following injury. These results suggest that a chronically progressive degenerative process may be initiated by brain trauma. Thus, there is a temporally broad window within which to introduce novel therapeutic strategies designed to ameliorate the short and long-term consequences of brain trauma.

Animals↗
PubMed1997

[Effect of zuogui wan on the contents of hypothalamic monoaminic transmitters and body weight in rats treated with monosodium glutamate neonatally].

OBJECTIVE: To evaluate the interrelationship between the traditional Chinese medicine the Kidney-Yin Deficiency Syndrome and the metabolism of hypothalamic monoaminic neurotransmitters after the lesion of arcuate nucleus (ARC). METHODS: Rats were injected with monosodium glutamate (MSG, at 2, 4, 6, 8, 10 days after born. 4 mg/g body weight, s.c.) during the neonatal period, then the contents of hypothalamic monoaminic transmitters were determined with HPLC and the changes of increase of body weight were observed. RESULTS: Decreases in levels of norepinephrine (NE), dopamine (DA) associated with its metabolite, 3,4-dihydroxyphenyl acetic acid (DOPAC) were observed, while vanillylmandelic acid (VMA), metabolite of NE, serotonin (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) were unchanged. Furthermore, adult rats with neonatal MSG treatment also demonstrated growth retardation, obesity but lower body weight, and shorter length of naso-anal. Zuogui Wan, classical drugs of nourishing and recuperating the Kidney-Yin, relieved the disorder of the metabolism of hypothalamic monoaminic transmitter in MSG-rats. CONCLUSION: The metabolic disorders of the central monoaminic transmitters may be related with the Deficiency of the Kidney-Yin.

Animals↗
PubMed1997

[Preparation and anti-HIV activity study of baicalien and its benzylated derivatives].

Baicalein (1) was prepared from baicalin by acid hydrolysis with yield of 24%. 5, 6-Dibenzyl-7-hydroxyl-flanone (4) and 6-benzyl-5, 7-dihydroxyl-flanone (5) were obtained by using different benzylating agents, and then hydrolyzing those benzylated compounds. The total yield of (4) and (5) were 5.9% and 5%, respectively. Both 4 and 5 were less effective than baicalien on inhibition of HIV-1 RT in vitro.

Anti-HIV Agents↗
PubMed1997

Characterization of diffuse axonal pathology and selective hippocampal damage following inertial brain trauma in the pig.

Dynamic deformation applied to white matter tracts is a common feature of human brain trauma, and may result in diffuse axonal injury (DAI). To produce DAI in an experimental model, we have utilized nonimpact inertial loading to induce brain trauma in miniature swine. This species was chosen due to its large gyrencephalic brain with substantial white matter domains. Twenty anesthetized (2% isoflurane) miniature swine were subjected to pure impulsive centroidal rotation 110 degrees in the coronal plane in 4 to 6 ms; peak accelerations ranged from 0.6 to 1.7 x 10(5) rad/s2. Seven days following injury, the brains were fixed (4% paraformaldehyde). Histopathologic examination was performed on 40 microns sections stained with cresyl violet (Nissl), antibodies targeting neurofilament (axonal damage), GFAP (astrocytes), and pig IgG (protein extravasation). Widespread multifocal axonal injury was observed in combination with gliosis throughout the brain, most commonly in the root of gyri and at the interface of the gray and white matter. Very little vascular disruption was noted in regions of axonal injury. Neuronal damage was primarily found in the CA1 and CA3 subfields of the hippocampus. These results suggest that this model is clinically relevant and useful for evaluating mechanisms of inertial brain trauma.

Animals↗
PubMed1996

Antisense confirmation of mu- and kappa-opioid receptor mediation of morphine's effects on body temperature in rats.

Previous studies showed that parenterally administered morphine at 4-16 mg/kg markedly increased body temperature in the rat, but higher doses of morphine (> or = 30 mg/kg, subcutaneously, sc) caused a profound decrease in body temperature. Based on the use of selective opioid agonists and antagonists, we postulated that these effects were due to morphine's actions on mu and kappa receptors, respectively. In the present study, we sought to determine whether an antisense (AS) oligodeoxynucleotide (oligo) against cloned mu or kappa opioid receptors could affect morphine-induced body temperature changes. AS oligos were directed against nucleotides 1-18 of the coding region of the mu receptor and 4-21 of the coding region of the kappa receptor. Male SD rats were surgically implanted with intracerebroventricular (icv) cannulae. Rats received icv injections of vehicle or oligo in the animal colony room on days 1, 3 and 5. Either AS oligo or missense (MS) oligo was infused in a volume of 5 microliters over 30 s to freely moving animals. On day 6, the rats were tested. The results showed that icv treatment with an AS oligo against mu opioid receptors, but not an MS oligo against the mu opioid receptor or an AS oligo against the kappa opioid receptor, significantly attenuated the hyperthermia normally produced by a relatively low dose of morphine administered sc. In addition, treatment with an AS oligo against kappa receptors, but not an MS oligo against kappa opioid receptor or an AS oligo against the mu opioid receptor, significantly blocked the hypothermia induced by a high dose of morphine. This study confirms our earlier postulate that morphine at 4 mg/kg, sc, induces an increase in body temperature primarily via mu opioid receptors in the brain and a high dose (30 mg/kg) of morphine administered sc produces a decrease primarily through kappa opioid receptors in the brain.

Animals↗
PubMed1996

Identification of tyrosine 187 as a protein kinase C-delta phosphorylation site.

Protein kinase C-delta (PKC-delta) has been demonstrated to be phosphorylated on tyrosine residue(s) in many different biological systems (Li, W., Yu, J.-C., Michieli, P., Beeler, J. F., Ellmore, N., Heidaran, M. A., and Pierce, J. H. (1994) Mol. Cell. Biol. 14, 6727-6735; Li, W., Mischak, H., Yu, J.-C., Wang, L.-M., Mushinski, J. F., Heidaran, M. A., and Pierce, J. H. (1994) J. Biol. Chem. 269, 2349-2352; Denning, M. F., Dlugosz, A. A., Howett, M. A., and Yuspa, S. H. (1993) J. Biol. Chem. 268, 26079-26081). Tyrosine phosphorylation of PKC-delta has also been shown to occur in vitro when purified PKC-delta is coincubated with different tyrosine kinase sources. However, the tyrosine phosphorylation site(s) is currently unknown and the exact effect of this phosphorylation on its serine/threonine kinase activity and biological functions is still controversial. To directly investigate the potential role of PKC-delta tyrosine phosphorylation, tyrosine 187 was converted to phenylalanine (PKC-deltaY187F) by site-directed mutagenesis, and expression vectors containing PKC-deltaY187F cDNAs were transfected into both 32D myeloid progenitor cells and NIH 3T3 fibroblasts. The results showed that tyrosine 187 of PKC-delta became phosphorylated in vivo in response to 12-O-tetradecanoylphorbol-13-acetate stimulation or platelet-derived growth factor receptor activation. In vivo labeling and subsequent two-dimensional phosphopeptide analysis demonstrated that one phosphopeptide was absent in PKC-deltaY187F when compared to wild type PKC-delta, further substantiating that tyrosine 187 of PKC-delta is phosphorylated in vivo. Although the phosphotyrosine content of PKC-deltaY187F was reduced compared with PKC-deltaWT, the kinase activity of PKC-deltaY187F toward a PKC-delta substrate was not altered. Moreover, 12-O-tetradecanoylphorbol-13-acetate-mediated monocytic differentiation of 32D cells was not affected by expression of the PKC-deltaY187F mutant. Taken together, these results suggest that tyrosine phosphorylation of PKC-delta on 187 may not influence PKC-delta activation and known functions.

3T3 Cells↗
PubMed1996

Localization of sympathetic and parasympathetic neurons innervating pancreas and spleen in the cat.

The localization of sympathetic and parasympathetic neurons innervating the pancreas and spleen was studied in the cat utilizing retrograde transport of wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP). Injection of WGA-HRP into the pancreas resulted in retrograde labeling in the whole of the solar plexus, while injection of WGA-HRP into the spleen also resulted in heavy labeling in the celiac ganglia bilaterally. Only a few labeled neurons were distributed in the superior mesenteric ganglion. With respect to parasympathetic innervation, HRP-labeled pancreatic and splenic neurons were found throughout the rostrocaudal extent of the dorsal motor nucleus of the vagus nerve (DMV) bilaterally. Although pancreatic neurons in the DMV were mainly observed in limits rostral to the obex, splenic neurons were centered at the level of the obex.

Animals↗
PubMed1996

Use of a mu-antisense oligodeoxynucleotide as a mu opioid receptor noncompetitive antagonist in vivo.

We examined whether mu-antisense (AS) oligodeoxynucleotide (oligo) treatment can be used in a manner similar to the mu-selective irreversible antagonist beta-funaltrexamine (beta-FNA) for in vivo pharmacology. Rats were injected intracerebroventricularly (icv) with a mu-AS or a missense (MS) oligo on days 1, 3, 5, 7, and 9 and were tested for the antinociceptive effect of sc injection of morphine on days 2, 4, 6, 8, and 10 in the cold water tail-flick (CWT) test. In another set of experiments, rats were also tested for the antinociceptive action of morphine twenty-four hours after icv injection of beta-FNA. Both beta-FNA and mu-AS produced rightward shifts in the dose-effect curves of morphine. In addition, pretreatment with 2.5 micrograms or more of beta-FNA or the mu-AS oligo for 5-9 days (but not for 1-3 days) reduced the maximal analgesic effect of morphine. The approximate fraction of functional receptor remaining for morphine was determined with the method of Furchgott to be 49.5% following 2.5 micrograms of beta-FNA; that after 5 days of the mu-AS oligo treatment was 50.8%. The results suggest that the mu-AS oligo can be used in the same manner as highly selective, irreversible mu opioid receptor ligands. Thus, properly designed AS oligos against receptors are of particular benefit when irreversible antagonists are not available. AS oligos represent a new class of selective and powerful pharmacological antagonists.

Animals↗
PubMed1996

Molecular basis of proton block of L-type Ca2+ channels.

Hydrogen ions are important regulators of ion flux through voltage-gated Ca2+ channels but their site of action has been controversial. To identify molecular determinants of proton block of L-type Ca2+ channels, we combined site-directed mutagenesis and unitary current recordings from wild-type (WT) and mutant L-type Ca2+ channels expressed in Xenopus oocytes. WT channels in 150 mM K+ displayed two conductance states, deprotonated (140 pS) and protonated (45 pS), as found previously in native L-type Ca2+ channels. Proton block was altered in a unique fashion by mutation of each of the four P-region glutamates (EI-EIV) that form the locus of high affinity Ca2+ interaction. Glu(E)-->Gln(Q) substitution in either repeats I or III abolished the high-conductance state, as if the titration site had become permanently protonated. While the EIQ mutant displayed only an approximately 40 pS conductance, the EIIIQ mutant showed the approximately 40 pS conductance plus additional pH-sensitive transitions to an even lower conductance level. The EIVQ mutant exhibited the same deprotonated and protonated conductance states as WT, but with an accelerated rate of deprotonation. The EIIQ mutant was unusual in exhibiting three conductance states (approximately 145, 102, 50 pS, respectively). Occupancy of the low conductance state increased with external acidification, albeit much higher proton concentration was required than for WT. In contrast, the equilibrium between medium and high conductance levels was apparently pH-insensitive. We concluded that the protonation site in L-type Ca2+ channels lies within the pore and is formed by a combination of conserved P-region glutamates in repeats I, II, and III, acting in concert. EIV lies to the cytoplasmic side of the site but exerts an additional stabilizing influence on protonation, most likely via electrostatic interaction. These findings are likely to hold for all voltage-gated Ca2+ channels and provide a simple molecular explanation for the modulatory effect of H+ ions on open channel flux and the competition between H+ ions and permeant divalent cations. The characteristics of H+ interactions advanced our picture of the functional interplay between P-region glutamates, with important implications for the mechanism of Ca2+ selectivity and permeation.

Animals↗
PubMed1996

Electrical stimulation at traditional acupuncture sites in periphery produces brain opioid-receptor-mediated antinociception in rats.

Previous studies in rats measuring latency to tail flick with radiant heat have shown that the antinociceptive effect induced by electrical stimulation of different frequencies at traditional acupuncture sites is mediated via different opioid receptors in the spinal cord. The present study was designed to observe (1) whether electrical stimulation at such sites could produce antinociceptive effects in the cold water tail-flick (CWT) test; (2) whether the antinociceptive effects could be blocked by s.c. injection of the opioid receptor antagonist naloxone and (3) whether i.c.v. injection of selective antagonists for mu (cyclic D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2, CTAP), delta (naltrindole) or kappa (nor-binaltorphimine) opioid receptors would block the antinociceptive effect produced by electrical stimulation. Sprague-Dawley rats were stimulated at frequencies of 2, 30 or 100 Hz with acupuncture needles inserted into acupoints Zusanli and Sanyinjiao in the hind leg for 30 min. Antinociception was assayed in the CWT. The results showed that (1) a significant, frequency-related increase in threshold in the CWT was observed in all electrical stimulation groups as compared with the placebo group and the antinociceptive effect lasted about 30 min poststimulation; (2) naloxone (s.c.) antagonized the antinociceptive effect induced by 2 Hz, 30 Hz or 100 Hz electrical stimulation and (3) either CTAP or naltrindole (i.c.v.) almost completely blocked the antinociceptive effect induced by 2 Hz or 30 Hz electrical stimulation, but was less effective in blocking antinociception induced by 100 Hz electrical stimulation; nor-binaltorphimine (i.c.v.) greatly reduced antinociception induced by 30 Hz or 100 Hz electrical stimulation, but not by 2 Hz electrical stimulation. These results indicate that the antinociception induced by 2 Hz electrical stimulation is mediated by both mu and delta opioid receptors; the antinociception induced by 100 Hz electrical stimulation is mediated primarily by the kappa receptor; and the antinociception induced by 30 Hz electrical stimulation is mediated by all three opioid receptor types. Thus, the antinociceptive effect induced by peripheral electrical stimulation, as measured by the CWT, involves opioid receptors in the rat brain.

Acupuncture Therapy↗
PubMed1996

[Effect of aconitine on content of corticotropin-releasing hormone in hypothalamus of rats].

The effect of aconitine on the content of corticotropin-Releasing-Hormone (CRH) in the hypothalamus of the normal rat injected with aconitine intraperitoneally was studied by means of radioimmunoassay (RIA). Aconitine is the main effective component of Radix Aconiti Carmichaeli Praeparata, the representative traditional Chinese herbal medicine of Warm-Supplementing Kidney-Yang (WSKY). Results showed that the content of hypothalamic CRH menifested a dosedependent increase after 7 days intraperitoneal injection of aconitine (1 microgram/kg, 3 micrograms/kg and 10 micrograms/kg). The CRH neurons in the paraventricular nucleus of hypothalamus and the neural fibers of median eminence also demonstrated increases in number and enrichment in gray tone as observed by immunohistochemical method of ABC. These results suggested that one of the possible mechanism of the WSKY drugs in improving the HPA axis might be exciting the hypothalamic CRH neurons.

Aconitine↗