Biomedical subjects

Q Zhou

Publications and source records attributed to Q Zhou.

At least 91 records · Page 5Linked to original sources

PubMed2001

Metabotropic glutamate receptor activation causes a rapid redistribution of AMPA receptors.

Electrophysiology, immunostaining and time lapse imaging techniques were employed to study the mechanism of long-term depression (LTD) induced by DHPG, a specific group I metabotropic glutamate receptor (mGluR) agonist. Experiments were performed in primary hippocampal culture or in the CA1 area of acute rat hippocampal slices. In agreement with previous results by others, we show that DHPG (200 microM, 10 min) can induce LTD (DHPG-LTD) in acute slices, in the presence or absence of synaptic inhibition. In addition, in voltage clamp whole cell experiments we find that accompanying the reduction in the evoked excitatory postsynaptic current (EPSC), miniature EPSC amplitude and frequency are reduced. Similar results were obtained in cultured neurons. Immunostaining and time lapse imaging showed a long-lasting loss of AMPA receptors from the membrane surface of cultured neurons after DHPG treatment, which appears to occur in only a subset of the puncta. Further electrophysiological recordings on slices showed that blocking postsynaptic endocytosis by introducing a blocking peptide named D15 in recording pipettes abolished the DHPG-LTD. In conclusion, these data suggest that LTD induced by mGluR activation is due to a rapid removal of AMPA receptors from the postsynaptic membrane.

Animals↗

PubMed2001

Evaluation of phosphorus bioavailability in sediments of three contrasting lakes in China and the UK.

The bioavailability of phosphorus (P) in sediments from West Lake and Lake Tai (China) and Lough Erne (Northern Ireland) was evaluated using total P (TP), water soluble P (WSP), readily desorbable P (RDP), algal available P (AAP) and Olsen-P. The results indicated that the TP content, the amounts of phosphorus extracted by each method and their proportions to TP varied widely between sediments of the three lakes. TP and chemical extractable phosphorus in the sediments of Lough Erne were considerably higher than those of the two lakes in China. Although TP in the West Lake sediments was similar to that of Lake Tai, the extractable phosphorus data suggested that the bioavailability of P in West Lake sediments was higher than that in sediments from Lake Tai. Nevertheless, the rank order of extraction efficiency was the same in all lakes and the amount extracted was in the sequence AAP > Olsen-P > WSP > RDP. There was a good correlation between extracted and TP in all the methods examined.

Biological Availability↗

PubMed2001

Identification and determination of nucleosides in rat brain microdialysates by liquid chromatography/electrospray tandem mass spectrometry.

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed for the determination of brain basal nucleosides (inosine, guanosine and adenosine) in microdialysates from the striatum and cortex of freely moving rats. A microdialysis probe was surgically implanted into the striatum or cortex of individual rats and Ringer's solution was used as the perfusion medium at a flow rate of 0.3 or 0.5 microl/min. The samples were then analyzed off-line by LC/MS/MS experiments. The separation of inosine, guanosine and adenosine was carried out on a cyano column using a mobile phase of 10 mM ammonium acetate, 1% acetic acid and 8% methanol at a flow rate of 0.4 ml/min. Analytes were detected by electrospray ionization tandem mass spectrometry in the positive ion mode. The detection limit for inosine, guanosine and adenosine was 80, 80 and 40 pg on column, respectively. With this method, the intercellular basal inosine, guanosine and adenosine concentrations in striatum and cortex of rat were determined.

Animals↗

PubMed2001

Sequence-specific delivery of a quinone methide intermediate to the major groove of DNA.

Silyl-protected phenol derivatives serve as convenient precursors for generating highly electrophilic quinone methide intermediates under biological conditions. Reaction is initiated by addition of fluoride and has previously exhibited proficiency in DNA alkylation and cross-linking. This approach has now been extended to the modification of duplex DNA through triplex recognition and fluoride-dependent quinone methide induction. Both oligonucleotides of a model duplex were alkylated in a sequence specific manner by an oligonucleotide conjugate that is consistent with triplex association. Optimum reaction required the presence of the two complementary target sequences and a pH of below 6.5. In addition, one guanine in each strand adjacent to the triplex region was the predominant site of alkylation. The yield of modification varied from approximately 20% for the purine-rich strand to only 4% for the pyrimidine-rich strand. This surprising difference indicates that the linker between the recognition and reactive elements may limit productive interaction between the quinone methide and the reactive nucleophiles of DNA. Restricted orientation of this intermediate may also be responsible for the lack of target cross-linking at detectable levels.

Alkylation↗

PubMed2001

Detecting AMD with multiply scattered light tomography.

PURPOSE: to use a novel technique, Multiply Scattered Light Tomography (MSLT), to provide a comfortable, rapid, and noninvasive method for detection and management of Age-related Macualar Degeneration. METHODS: two patient groups were studied in clinical settings with MSLT and confocal scanning laser tomography. In Poway, CA, 21 retinal patients underwent tomography, and the 17 patients with suspicion of exudation also had ICG. An Angio-Scan (Laser Diagnostic Technologies, Inc.) was used to provide simultaneous fundus reflectance and ICG imaging. In Methuen, MA, 20 retinal patients underwent tomography with fluorescein angiography for suspicion of exudation. The MSLT was based on the TopSS (Laser Diagnostic Technologies, Inc.), with a Vertical Cavity Surface Emitting Laser array at 850 mm as the illumination source. The central laser produced confocal images. The surrounding lasers produced multiply scattered light images. RESULTS: MSLT emphasized structures beneath the retina such as drusen, choroidal new vessel membranes, and pigment epithelial detachments. Exudation seen on angiography was visualized by MSLT as topographical structures with distinct borders. Superficial structures, e.g., cysts and epiretinal membranes, were visualized in 850 nm images. DISCUSSION: confocal tomography and MSLT provided a rapid, noninvasive method to detect and localize macular degeneration and pathological structures found in eyes of older patients.

Adult↗

PubMed2001

Estrogens and epididymal function.

Estrogen is synthesized in the male reproductive system and is found in high concentrations in rete testis and seminal fluids. This luminal estrogen targets estrogen receptors (ER) along the male reproductive tract, and in particular the efferent ductules, where ERalpha is abundant. However, both ERalpha and ERbeta are found in various regions of the male reproductive tract. The transgenic ER knockout mice (alphaERKO and betaERKO) have been used to help define the role of ER in the male. In the alphaERKO animal model, the efferent ductules are dramatically altered, forming an epithelium in which fluid reabsorption is inhibited and epithelial cells have greatly reduced numbers of lysosomes and organelles associated with endocytosis. The betaERKO male reproductive tract appears normal. Because these animals are transgenic and lack ER throughout development, we developed animal models using pure antiestrogen ICI 182,780 treatments in adult males. The data show that ERalpha participates in the regulation of the apical cytoplasm of non-ciliated cells of the efferent ductules, narrow cells of initial segment epididymis and clear cells in the remaining segments of the epididymis. There appears to be no effect on vas deferens. The inhibition of ERalpha function in the male leads to decreases in sperm concentrations and eventually to infertility. The current literature leaves the mechanisms of estrogen action in the male reproductive tract unsettled and raises the question of androgen's contribution to the regulation of fluid transport, especially in the efferent ductules.

Animals↗

PubMed2001

Developmental potential of mouse embryos reconstructed from metaphase embryonic stem cell nuclei.

Mice have recently been successfully cloned from embryonic stem (ES) cells. However, these fast dividing cells provide a heterogeneous population of donor nuclei, in terms of cell cycle stage. Here we used metaphases as a source of donor nuclei because they offer the advantage of being both unambiguously recognizable and synchronous with the recipient metaphase II oocyte. We showed that metaphases from ES cells can provide a significantly higher development rate to the morula or blastocyst stage (56--70%) than interphasic nuclei (up to 28%) following injection into a recipient oocyte. Selective detachment of mitotic cells after a demecolcin treatment greatly facilitates and accelerates the reconstruction of embryos by providing a nearly pure population of cells in metaphase and did not markedly affect the developmental rate. Most of the blastocysts obtained by this procedure were normal in terms of both morphology and ratio of inner cell mass and total cell number. After transfer into pseudopregnant recipients at the one- or two-cell stage, the ability of metaphase to be fully reprogrammed was demonstrated by the birth of two pups (1.5% of activated oocytes). Although the implantation rate was quite high (up to 32.9% of activated oocytes), the postimplantation development was characterized by a high and rapid mortality. Our data provide a clear situation to explore the long-lasting effects that can be induced by early reprogramming events.

Animals↗

PubMed2001

Insertional mutagenesis of genes required for seed development in Arabidopsis thaliana.

The purpose of this project was to identify large numbers of Arabidopsis genes with essential functions during seed development. More than 120,000 T-DNA insertion lines were generated following Agrobacterium-mediated transformation. Transgenic plants were screened for defective seeds and putative mutants were subjected to detailed analysis in subsequent generations. Plasmid rescue and TAIL-PCR were used to recover plant sequences flanking insertion sites in tagged mutants. More than 4200 mutants with a wide range of seed phenotypes were identified. Over 1700 of these mutants were analyzed in detail. The 350 tagged embryo-defective (emb) mutants identified to date represent a significant advance toward saturation mutagenesis of EMB genes in Arabidopsis. Plant sequences adjacent to T-DNA borders in mutants with confirmed insertion sites were used to map genome locations and establish tentative identities for 167 EMB genes with diverse biological functions. The frequency of duplicate mutant alleles recovered is consistent with a relatively small number of essential (EMB) genes with nonredundant functions during seed development. Other functions critical to seed development in Arabidopsis may be protected from deleterious mutations by extensive genome duplications.

Alleles↗

PubMed2001

Arabidopsis genes essential for seedling viability: isolation of insertional mutants and molecular cloning.

We have undertaken a large-scale genetic screen to identify genes with a seedling-lethal mutant phenotype. From screening approximately 38,000 insertional mutant lines, we identified >500 seedling-lethal mutants, completed cosegregation analysis of the insertion and the lethal phenotype for >200 mutants, molecularly characterized 54 mutants, and provided a detailed description for 22 of them. Most of the seedling-lethal mutants seem to affect chloroplast function because they display altered pigmentation and affect genes encoding proteins predicted to have chloroplast localization. Although a high level of functional redundancy in Arabidopsis might be expected because 65% of genes are members of gene families, we found that 41% of the essential genes found in this study are members of Arabidopsis gene families. In addition, we isolated several interesting classes of mutants and genes. We found three mutants in the recently discovered nonmevalonate isoprenoid biosynthetic pathway and mutants disrupting genes similar to Tic40 and tatC, which are likely to be involved in chloroplast protein translocation. Finally, we directly compared T-DNA and Ac/Ds transposon mutagenesis methods in Arabidopsis on a genome scale. In each population, we found only about one-third of the insertion mutations cosegregated with a mutant phenotype.

Arabidopsis↗

PubMed2001

Gene and protein expressions of nitric oxide synthases in ischemia-reperfused peripheral nerve of the rat.

This study examined mRNA and protein expressions of neuronal (nNOS), inducible (iNOS), and endothelial nitric oxide synthases (eNOS) in peripheral nerve after ischemia-reperfusion (I/R). Sixty-six rats were divided into the ischemia only and I/R groups. One sciatic nerve of each animal was used as the experimental side and the opposite untreated nerve as the control. mRNA levels in the nerve were quantitatively measured by competitive PCR, and protein was determined by Western blotting and immunohistochemical staining. The results showed that, after ischemia (2 h), both nNOS and eNOS protein expressions decreased. After I/R (2 h of ischemia followed by 3 h of reperfusion), expression of both nNOS and eNOS mRNA and protein decreased further. In contrast, iNOS mRNA significantly increased after ischemia and was further upregulated (14-fold) after I/R, while iNOS protein was not detected. The results reveal the dynamic expression of individual NOS isoforms during the course of I/R injury. An understanding of this modulation on a cellular and molecular level may lead to understanding the mechanisms of I/R injury and to methods of ameliorating peripheral nerve injury.

Animals↗

PubMed2001

Comprehensive-trace level determination of methyltin compounds in aqueous samples by cryogenic purge-and-trap gas chromatography with flame photometric detection.

A comprehensive method was developed for the sensitive and fast determination of trace levels of methyltin compounds in aqueous samples. Tin compounds in aqueous solution at pH 5 were converted to the corresponding volatile hydrides: CH3SnH3, (CH3)2SnH2, and (CH3)3SnH, by reaction with potassium borohydride. A CP-4010 purge and trap injector (PTI) was used to purge analyte species from water directly. The volatile derivatives were base-line separated on a capillary column in an Angilent-6890 gas chromatograph by a suitable temperature program and were detected by a flame photometric detector (FPD). The detection limits were 18 ng L-1 for monomethyltin, 12 ng L-1 for dimethyltin, and 3 ng L-1 for trimethyltin, respectively. This method was successfully applied to the determination of methyltin compounds in different aqueous samples.

Chemical Industry↗

PubMed2001

Contortrostatin, a homodimeric disintegrin, actively disrupts focal adhesion and cytoskeletal structure and inhibits cell motility through a novel mechanism.

Integrins play a major role in the regulation of cell motility. They physically link the extracellular environment to the cytoskeleton and participate in large protein complexes known as focal adhesions. In this report, it is demonstrated that treatment of tumor cells with the homodimeric disintegrin contortrostatin induces integrin-mediated tyrosine phosphorylation events and causes severe disruptions in the actin cytoskeleton and disassembly of focal adhesion structures without affecting cellular adhesion to a reconstituted basement membrane. Included in this disruption is the tyrosine phosphorylation and altered subcellular localization of FAK. Through use of transfected 293 cells with specific integrin expression profiles and anti-alphavbeta3 mAbs, we demonstrate that these events are mediated exclusively by the alphavbeta3 integrin and are likely the result of contortrostatin-mediated crosslinking of this receptor at the cell surface, since monovalent disintegrins, flavoridin or echistatin do not induce such effects. Further, it is shown that contortrostatin potently inhibits motility in cells expressing the alphavbeta33 integrin. The results of this study describe a novel integrin-mediated mechanism by which cell motility can be inhibited and suggest an alternative approach to therapeutic intervention for cancer invasion and metastasis.

Animals↗

PubMed2001

Chiral metabolism of propafenone in rat hepatic microsomes treated with two inducers.

AIM: To study the influence of inducers of drug metabolism enzyme, beta-naphthoflavone (BNF) and dexamethasone (DEX), on the stereoselective metabolism of propafenone in the rat hepatic microsomes. METHODS: Phase I metabolism of propafenone was studied using the microsomes induced by BNF and DEX and the non-induced microsome was used as the control. The enzymatic kinetics parameters of propafenone enantiomers were calculated by regress analysis of Eadie-Hofstee Plots. Propafenone enantiomer concentrations were assayed by a chiral HPLC. RESULTS: The metabolite of propafenone, N-desalkylpropafenone, was found after incubation of propafenone with the rat hepatic microsomes induced by BNF and DEX. In these two groups, the stereoselectivity favoring R(-) isomer was observed in metabolism at low substrate concentrations of racemic propafenone, but lost the stereoselectivity at high substrate concentrations. However, in control group, no stereoselectivity was observed. The enzyme kinetic parameters were: (1) K(m). CONTROL GROUP: R(-) 83+/-6, S(+) 94+/-7; BNF group: R(-) 105+/-6, S(+)128+/-14; DEX group: R(-) 86+/-11, S(+) 118+/-16; (2)V(max). CONTROL GROUP: R(-) 0.75+/-0.16, S(+) 0.72+/-0.07; BNF group: R(-) 1.04+/-0.15, S(+)1.07+/-14; DEX group: R(-) 0.93+/-0.06, S(+) 1.04+/-0.09; (3)Cl(int). CONTROL GROUP: R(-) 8.9+/-1.1, S(+) 7.6+/-0.7; BNF group: R(-) 9.9+/-0.9, S(+)8.3+/-0.7; DEX group: R(-) 10.9+/-0.8, S(+) 8.9+/-0.9. The enantiomeric differences in K(m) and Cl(int) were both significant, but not in V(max), in BNF and DEX group. Whereas enantiomeric differences in three parameters were all insignificant in control group. Furthermore, K(m) and V(max) were both significantly less than those in BNF or DEX group. In the rat liver microsome induced by DEX, nimodipine (NDP) decreased the stereoselectivity in propafenone metabolism at low substrate concentration. The inhibition of NDP on the metabolism of propafenone was stereoselective with R(-)-isomer being impaired more than S(+)-isomer. The inhibition constant (Ki) of S(+)- and R(-)-propafenone, calculated from Dixon plots, was 15.4 and 8.6 mg x L(-1), respectively. CONCLUSION: CYP1A subfamily(induced by BNF) and CYP3A4 (induced by DEX) have pronounced contribution to propafenone N-desalkylation which exhibited stereoselectivity depending on substrate concentration. The molecular base for this phenomenon is the stereoselectivity in affinity of substrate to the enzyme activity centers instead of at the catalyzing sites.

Animals↗

PubMed2001

Preliminary research on myosin light chain kinase in rabbit liver.

AIM: To study preliminarily the properties of myosin light chain kinase (MLCK) in rabbit liver. METHODS: The expression of MLCK was detected by reverse transcription-polymerase chain reaction(RT-PCR); the MLCK was obtained from rabbit liver, and its activity was analyzed by gamma-(32)P incorporation technique to detect the phosphorylation of myosin light chain. RESULTS: MLCK was expressed in rabbit liver, and the activity of the enzyme was similar to rabbit smooth muscle MLCK, and calmodulin-dependent. When the concentration was 0.65 mg x L(-1), the activity was at the highest level. CONCLUSION: MLCK expressed in rabbit liver may catalyze the phosphorylation of myosin light chain, which may play important roles in the regulation of hepatic cell functions.

Animals↗

PubMed2001

[Reduction of FHIT gene expression in primary lung cancer: relationship with the proliferation and apoptosis of tumor cell].

OBJECTIVE: To investigate the role of FHIT(fragile histidine triad) gene in oncogenesis and progression of human lung cancer and explore the relationship of FHIT gene expression with the proliferation and apoptosis of tumor cells. METHODS: The expression of FHIT gene and that of Ki-67 were detected in 166 lung cancer samples and 37 lung benign lesion tissues as control by immunohistochemistry, and the apoptotic level of tumor cell was detected by terminal deoxynucl neotidyl transferase mediated dUTP nick end labelling(TUNEL). RESULTS: (1)The expression level of FHIT gene was found to be significantly lower in lung cancer tissues than in benign lesion tissues (P<0.01); (2)The expression level of FHIT gene was closely related to histological classification, cancer cell differentiation, P-TNM stages and lymph node involvement in lung cancer patients (P<0.05); (3)Lasting and heavy smoking might be one of the important reasons of reduction of FHIT gene expression in lung cancer patients; (4)The postoperative survival time of patients in high FHIT expression group was significantly longer than those in low expression group (P<0.05); (5)The reduction of FHIT gene expression might be related with the over-proliferation and suppression of apoptosis of lung cancer cell(P<0.01). CONCLUSION: FHIT gene may play a role in the regulation of proliferation and apoptosis of lung cancer cell. And reduction of FHIT gene expression may be related to the pathogenesis and development of lung cancer.

Acid Anhydride Hydrolases↗

PubMed2001

Parasympathetic innervation of the meibomian glands in rats.

PURPOSE: To determine the location of parasympathetic neurons that innervate the meibomian glands in rats. METHODS: The B subunit of cholera toxin (CTB), fast blue, and a retrograde transneuronal tracer, the Bartha strain of pseudorabies virus (PRV-Ba), were injected into the upper eyelids of adult Sprague-Dawley rats after sectioning the ipsilateral branches of the facial nerve and resecting the superior cervical ganglia. Brains and orbital tissues were processed for the immunohistochemical detection of PRV-Ba and CTB. In selected cases, series of brain sections were double labeled for PRV-Ba and tyrosine hydroxylase to determine the relationship between the A5 noradrenergic cell group and superior salivatory nucleus, or for PRV-Ba and choline acetyltransferase to establish the neurochemical phenotype of parasympathetic preganglionic neurons. RESULTS: Labeled ganglionic cells were diffusely distributed within the ipsilateral pterygopalatine ganglion (PPG) and along the more proximal portions of the greater petrosal nerve (GPN). Labeled preganglionic neurons were cholinergic and were located immediately dorsolateral to the rostral-most portion of the facial nucleus and caudal superior olive, where they intermingled with A5 noradrenergic cells. CONCLUSIONS: The meibomian glands and other structures within the lid margin are subject to parasympathetic regulation by ganglion cells diffusely distributed within the PPG and along more proximal portions of the GPN. Cholinergic parasympathetic preganglionic neurons that project to meibomian gland-innervating ganglion cells are located immediately lateral, dorsal, and rostral to the facial motor nucleus in the region commonly referred to as the superior salivatory nucleus.

Amidines↗

PubMed2001

Stereoselective metabolism of propafenone by human liver CYP3A4 expressed in transgenic Chinese hamster CHL cells lines.

AIM: To investigate the stereoselective metabolism of propafenone (PPF) by human liver CYP3A4. METHODS: A chiral and an achiral HPLC were combined to determine the enantiomer of PPF in S9 incubates prepared from transgenic Chinese hamster CHL cells lines expressing CYP3A4. The time-dependent study was performed using individual enantiomer or racemate at low or high substrate concentration. Kinetic parameters were determined employing individual enantiomers as substrates. Enantiomeric inhibition experiments were performed by using R(-)-PPF as an inhibitor and S(+)-PPF as a substrate. RESULTS: Stereoselectivity was found in metabolism of racemic PPF at low substrate concentration (10 mg/L) (S < R), and lost at high substrate concentration (400 mg/L) When an individual enantiomer of high concentration (200 mg/L) was used as a substrate, S(+)-PPF was eliminated faster than its isomer (S < R). However, the opposite situation was observed at low concentration (5 mg/L) (S < R). The Vmax of S(+)-PPF was significantly greater than that of R(-)-PPF [(2.66 +/- 0.32) vs (1.71 +/- 0.19) micromol . mg-1 . min-1]. The Km of R(-)-PPF was significantly lower than that of S(+)-PPF [(73 +/- 11) vs (185 +/- 17) micromol/L]. R(-)-PPF inhibited S(+)-isomer with an IC50 value of 125 micromol . L-1. CONCLUSION: It is concluded that stereoselectivity in metabolism of propafenone via CYP3A4 depend on substrate concentration. Enantiomer/enantiomer interaction of PPF occurred at high concentration of substrate, and resulted in the loss of stereoselectivity. There maybe no enantiomer/enantiomer interaction at low concentration thus keeping the superiority of R(-)-PPF in metabolism.

Animals↗

PubMed2001

[Spatial structure and pattern of Nilaparvata lugens population in large-scale].

Brown planthopper(BPH), Nilaparvata lugens, is a major pest of rice. Its spatial distribution of different generations in Guangdong Province was studied by using variograms. The results showed that this population distributed in a clump pattern in rice fields during the 1st rice season. 3 generations of this population had different spatial structures, with the clump range of 400 km, 200 km and 205 km respectively in the 1st rice season. The simulation distribution maps of BPH, which was interpolated by Ordinary Kriging, showed that the density of BPH populations in west Guangdong province were higher than that in the east. The effect of sampling size on the spatial structure of BPH was not significant.

Animals↗