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Biomedical subjects

Q H Li

Publications and source records attributed to Q H Li.

At least 19 recordsLinked to original sources

Protein encoded by HSV-1 stimulation-related gene 1 (HSRG1) interacts with and inhibits SV40 large T antigen.

Herpes simplex virus (HSV)-1 stimulation-related gene 1 (HSRG1) protein expression is induced in HSV-1 infected cells. We found that HSRG1 interacts with SV40 large T antigen (LT) in yeast two-hybrid assay and bimolecular fluorescence complementation (BiFC) assay. This interaction alters LT's regulation of the SV40 promoter and its ability to influence the cell cycle. Choramphenicol acetyl-transferase (CAT) assays revealed that initiation of gene transcription by LT is changed by HSRG1 expression. HSRG1 inhibits the ability of LT to activate SV40 late gene transcription. Further data indicate that the ability of LT protein to stimulate S-phase entry is also inhibited by the expression of HSRG1. The results of a colony-forming assay suggested that expression of HSRG1 in cells transfected by LT gene decreased the rate of colony formation. Yeast two-hybrid beta-galactosidase assay revealed that amino acid residues 132-450 in LT bind HSRG1.

Animals↗

Interactive image-guided management of colloid cysts of the third ventricle.

OBJECT: Image guidance provides a three-dimensional view of the lesion and allows the surgeon to plan a surgical strategy that takes the relationship of the lesion and the surrounding brain into account. We evaluated the degree of resection and the functional outcome of patients with colloid cysts from the third ventricle submitted to surgical resection using interactive image-guided approach. METHOD: Using image-guided methodology and an endoscopic approach we analyzed the functional outcome of 11 patients with diagnosis of colloid cyst of the third ventricle who were treated at our institution from August 1993 to September 2000. The mean age was 39.5 years and the mean follow-up was 36.5 months. Analyzing the clinical outcome, 54.5% of the patients developed short-term memory disturbance in the first 30 days after surgery. None of these patients persisted with this symptomatology for more than one month. In terms of late post-operative morbidity, 1 patient developed persistent post-operative seizures, which were controlled with anti-seizure medications. Complete resection of the cyst was achieved in all patients. CONCLUSIONS: The low rate of complications and high rate of total resection encourage us to continue using the multimodal technique. Longer follow-up and an increase in the number of patients are needed to assess the efficacy of this methodology.

Adolescent↗

[Bibliometric analysis on tissue engineering research literatures].

OBJECTIVE: To comprehend the progress of tissue engineering research and speculate its developmental trends. METHODS: MEDLINE search was conducted to retrieve the papers published between 1987 to 1999 under the main headings of tissue engineering. Years, nationalities, languages, journals, authors and heading frequencies of 314 papers were analyzed by bibliometrics. RESULTS: Since 1990, the number of tissue engineering research literatures had doubled, and papers between 1998 and 1999 made up 57.96% of the total papers. All papers came from 15 nations, in 6 languages and 140 journals; 64.97% came from United States and 25.79% from England, Netherlands and Germany; 93.95% was in English; 42.04% was published on 15 journals. Vacanti JP and 19 other authors presented 5 to 24 papers. Heading frequencies were cytology 22.89%, transplantation 13.30%, scaffolds and extracellular matrixes 11.72%, implanting 10.60%, polymers 8.91%, potential applications 8.91%, artificial substitutes 6.88%, tissue culture 6.70% and biogenetics 4.96%. CONCLUSION: Tissue engineering literatures mainly come from United States, England, Netherlands and Germany. English is the major language. J Biomed Mater Res and 14 other journals are important journals about tissue engineering research. Vacanti JP and 19 other authors are prolific authors. Cytology, transplantation, scaffolds and extracellular matrixes and implanting are hot topics and key points on tissue engineering research.

Animals↗

Human CYP2C-mediated stereoselective phenytoin hydroxylation in Japanese: difference in chiral preference of CYP2C9 and CYP2C19.

Regio- and stereoselective hydroxylation of phenytoin was determined in liver microsomes of nine extensive (EM) and three poor metabolizers (PM) of mephenytoin. Hydroxyphenytoins (HPPH) were isolated and quantified after separation into four regio- and stereoisomers. The total rates of microsomal phenytoin 4'- hydroxylation were approximately 3-fold higher than those of 3'-hydroxylation, and not significantly different in EM and PM. Formation of 4'-(R)-HPPH was 4.4-fold higher in EM than in PM, whereas no clear differences between EM and PM were detected in the formation of 4'-(S)-, 3'-(R)-, and 3'-(S)-HPPH. Cytochrome P450 (CYP)2C9, expressed in a fission yeast, Schizosaccharomyces pombe, catalyzed the formation of 4'-(R)- and 4'-(S)-HPPH stereoselectively, as observed with EM, in which predominantly 4'-(S)-HPPH was formed. Recombinant CYP2C19 was more stereoselective for 4'-(R)-HPPH formation. These results, in addition to inhibition experiments with anti-human CYP2C antibody, indicate that phenytoin hydroxylation is mainly catalyzed by CYP2C9. Furthermore, CYP2C19 showed limited contribution to phenytoin 4'-hydroxylation with a different chiral preference from CYP2C9.

Antibodies↗

Comparative investigation on spindle behavior and MPF activity changes during oocyte maturation between gynogenetic and amphimictic crucian carp.

The spindle behavior and MPF activity changes in the progression of oocyte maturation were investigated and compared with cytological observation and kinase assay between gynogenetic silver crucian carp and amphimictic colored crucian carp. MPF activity was measured by using histone H1 as phosphorylation substrate. There were two similar oscillatory MPF kinase activity changes during oocyte maturation in two kinds of fishes with different reproductive modes, but there existed some subtle difference between them. The subtle difference was that the first peak of MPF kinase activity was kept to a longer-lasting time in the gynogenetic silver crucian carp than in the amphimictic colored crucian carp. It was suggested that the difference may be related to the spindle behavior changes, such as tripolar spindle formation and spindle rearrangement in the gynogenetic crucian carp.

Animals↗

Poliovirus mutants at histidine 195 of VP2 do not cleave VP0 into VP2 and VP4.

The final stage of poliovirus assembly is characterized by a cleavage of the capsid precursor protein VP0 into VP2 and VP4. This cleavage is thought to be autocatalytic and dependent on RNA encapsidation. Analysis of the poliovirus empty capsid structure has led to a mechanistic model for VP0 cleavage involving a conserved histidine residue that is present in the surrounding environment of the VP0 cleavage site. Histidine 195 of VP2 (2195H) is hypothesized to activate local water molecules, thus initiating a nucleophilic attack at the scissile bond. To test this hypothesis, 2195H mutants were constructed and their phenotypes were characterized. Consistent with the requirement of VP0 cleavage for poliovirus infectivity, all 2195H mutants were nonviable upon introduction of the mutant genomes into HeLa cells. Replacement of 2195H with threonine or arginine resulted in the assembly of a highly unstable 150S virus particle. Further analyses showed that these particles contain genomic RNA and uncleaved VP0, criteria associated with the provirion assembly intermediate. These data support the involvement of 2195H in mediating VP0 cleavage during the final stages of virus assembly.

Capsid↗

Use of the Wei-Lin-Weissfeld method for the analysis of a recurring and a terminating event.

We consider application of the Wei-Lin-Weissfeld (WLW) method for multiple failure time data when analysing a disease process consisting of a recurring outcome, such as clinical progression, and a terminating outcome, such as death. In order to adapt WLW for this situation, 'events' must be specified that define multiple failure times and whether these are censored. Various choices of events are possible, and each corresponds to inferences about a different aspect of the underlying disease process. Definitions which regard the terminating outcome as a censor of the recurring outcome focus on specific cause-specific hazard functions, while event definitions which make no distinction between a recurring and terminating outcome focus on hazard functions of the induced failure times. Some event definitions require strong statistical assumptions to yield valid inferences and are not recommended. The application of WLW for recurring/terminating processes is illustrated with the results of two recently conducted clinical trials in persons with HIV.

Acquired Immunodeficiency Syndrome↗

Application accuracy study of a semipermanent fiducial system for frameless stereotaxis.

The accuracy of a semipermanent fiducial marker system developed at Wayne State University in collaboration with Fisher-Leibinger (Freiburg, Germany) was compared with reference to a standard stereotactic frame (Zamorano-Dujovny Localizing Unit; Fisher-Leibinger). For each patient in our study, 10 semipermanent markers were placed on the skull through a small incision and a pilot hole drilled for the marker; five markers were used for registration, and five were used for comparison. Gadolinium-enhanced magnetic resonance imaging was performed, and, upon registration using both ring and fiducial markers, 184 random points were collected by infrared digitization. All three-dimensional measurements (x, y, z) were converted into distance values correlating each value to the origin by the formula dij = SQRT (xij2 + yij2 + zij2). The mean difference of fiducial coordinates vs. absolute image coordinates was 1.72 +/- 0.42 mm (P = .0001), implying no significant difference. The mean difference in dij of the stereotactic ring coordinates vs. the absolute image coordinates was 3.35 +/- 0.59 mm (P = .00011). The mean difference in the fiducial markers vs. the stereotactic ring coordinates was 2.95 +/- 0.45 mm (P = .0001). All tests were declared significant at alpha = .016. The combination of interactive guidance with semipermanent fiducial markers allows for accurate localization of intracranial targets (as accurate or even more accurate than the stereotactic frame). Semipermanent fiducial markers facilitate the procedure logistically, allow for staged procedures (i.e., at the skull base or in epilepsy), and provide access for combined supra- and infratentorial approaches. We believe that the semipermanent fiducial markers system might represent an important development leading toward widespread use of interactive image guidance in conventional neurosurgery.

Cerebellum↗

Vasodilator effect of human adrenomedullin(13-52) on hypertensive rats.

Human adrenomedullin (hADM) is a newly isolated peptide with hypotensive activity in normotensive rats. The objective of this study was to investigate the effect of hADM(13-52) on hypertensive animals. hADM(13-52) induced a dose-dependent decrease in the blood pressure of spontaneously hypertensive rats and renal hypertensive rats. This result suggests that hADM is a novel antihypertensive peptide. In isolated rat aortic arteries, hADM(13-52) produced nitric oxide dependent relaxation and inhibited endothelin 1 and angiotensin II release. These in vitro effects may represent the molecular mechanisms underlying the hypotensive action of hADM in vivo.

Adrenomedullin↗

[Membrane fusion between Ehrlich ascites of mastocarcinoma cells and liposome induced by proton translocation of transplasma membrane NADH-ferricyanide redox enzymes].

By using resonance energy transfer assay (RET) and fluorescence microscopy we show experimental evidence that membrane fusion of Ehrlich ascites of mastocarcinoma cells with liposomes could be induced by the proton translocation activity associated with NADH-ferricyanide redox enzyme of transplasma membrane of cancer cells. The iodoacetate, an inhibitor of glycolysis, was found to be able to depress the proton translocation activity and also to inhibit the membrane fusion. It is suggested that NADH produced mainly by glycolysis is utilized as the substrate (electron donor) for transmembrane ferricyanide reduction, and the proton pumping activity in the cancer cells is coupled to the transmembrane NADH-ferricyanide redox enzyme system. Experiments also show that membrane fusion extent of cancer cells with liposomes is proportional to the amount of H+ pumped out by the cells and membrane fusion process also exhibits a H+ consuming mode just as in fusion process of mitochondria with liposomes by redox enzyme proton pumps of respiratory chain. All of the results presented in this paper consists with recent reports of this laboratory, which indicated that various types of proton pumping system from different membrane system of cell have a new function in membrane fusion. Therefore, the proton pumping induced membrane fusion may have a more general physiological importance in triggering and modulating fusion process of native membrane in vivo.

Animals↗

Lack of low Km diazepam N-demethylase in livers of poor metabolizers for S-mephenytoin 4'-hydroxylation.

Metabolism of diazepam was studied in vitro to identify the forms of cytochrome P450 (CYP) responsible for N-demethylation (nordazepam formation) and 3-hydroxylation (temazepam formation), using liver microsomes obtained from extensive (EM) and poor metabolizers (PM) for S-mephenytoin 4'-hydroxylation. Involvement of at least two P450 forms in diazepam N-demethylation was suggested by a biphasic pattern in Lineweaver-Burk and Eadie-Hofstee plots from the EM, whereas a monophasic pattern was observed from the PM liver microsomes. The kinetic parameters for the N-demethylation in the EM group were: Km 1, 19.4 +/- 0.4 microM; Vmax 1, 0.27 +/- 0.04 nmol min-1 per mg protein; Km 2, 346 +/- 34 microM; Vmax2, 1.82 +/- 0.63 nmol min-1 per mg protein (n = 3, mean +/- SD). The PM group showed the mean values of Km and Vmax (Km, 319 +/- 30 microM; Vmax, 1.49 +/- 0.62 nmol min-1 per mg protein) (n = 3) similar to those of Km2 and Vmax2 in the EM group. An antibody raised against CYP2C9 (anti-human CYP2C) strongly inhibited diazepam N-demethylation in EM liver microsomes at a low substrate concentration (20 microM). However, the anti-human CYP2C showed no clear inhibition of N-demethylation in EM liver microsomes at a high substrate concentration (200 microM). Diazepam N-demethylation in PM liver microsomes was not clearly inhibited by the anti-human CYP2C at either the low or high substrate concentrations. These data suggest that different P450 forms mediated diazepam N-demethylation in EM and PM liver microsomes, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies↗

[Semiquantitative detection and application about the expression of nitric oxide synthase gene].

Using the Reverse Transcription (RT)-Polymerase Chain Reaction (PCR) method of Martin (1993) for semiquantitation determinating of NOS gene, it was found that NOS mRNA is not only existed in brain, but also distributed extensively in heart, kidney, lung and liver. Among of them, NOS mRNA levels were highest in the brain, followed in descending order by kidney, heart. In addition to endothelial cell, NOS gene was also highly expressed in smooth muscle cells, suggesting that they may be an important site of NOS in organism. Furthermore, NOS mRNA levels were found to decrease significantly in brain, kidney, liver and smooth muscle cell in spontaneous hypertensive rat. These data suggest that pathogeny of hypertension may be related to low expression of NOS gene in these tissues.

Amino Acid Oxidoreductases↗

Effects of K(+)-channel blockers on cochlear potentials in the guinea pig.

The effects of different K+ channel blockers, 4-aminopyridine (4-AP), tetraethylammonium (TEA) and quinine, on the various cochlear potentials were observed by the means of perilymph infusion. Each of the three blockers depressed the compound action potential. However, they exerted quite different effects on other cochlear potentials, especially comparing 4-AP, a fast K(+)-channel blocker, with two other blockers. 4-AP induced a significant increase in the magnitude of summating potential, while TEA and quinine decreased it; 4-AP showed no effect on the general endocochlear potential (G-EP, the EP value recorded directly from the scala media, SM) and the negative EP component (N-EP), while TEA and Quinine increased G-EP and decreased the absolute value of N-EP. They also exerted different effects on the EP changes induced by exposure to intense noise. The results indicate the different roles of different K(+)-channels in the generation of cochlear potentials. The relationship of the two components of EP (positive and negative) and the G-EP was discussed.

4-Aminopyridine↗

[Changes in endocochlear potential induced by potassium-channel blockers].

The effect of various potassium-channel blockers, 4-aminopyridine (4-AP), tetraethylammonium (TEA) and quinine, on the endocochlear potential (EP) was studied in perfused guinea pig inner ears. The fast K(+)-channel blocker, 4-AP, did not alter EP but changed its response model to intense noise exposure. While TEA and quinine significantly reduced the amplitude of negative component of EP (N-EP), comparing with a relatively smaller increase in general EP (G-EP). The results indicated the existence of different K(+)-channels with different physiological functions.

4-Aminopyridine↗

Purification and characterization of a developmentally regulated carboxypeptidase from Mucor racemosus.

A developmentally regulated carboxypeptidase was purified from hyphae of the dimorphic fungus Mucor racemosus. The enzyme, designated carboxypeptidase 3 (CP3), has been purified greater than 900-fold to homogeneity and characterized. The carboxypeptidase migrated as a single electrophoretic band in isoelectric focusing polyacrylamide gel electrophoresis (PAGE), with an isoelectric point of pH 4.4. The apparent molecular mass of the native enzyme was estimated by gel filtration to be 52 kDa. Sodium dodecyl sulfate (SDS)-PAGE under nonreducing conditions revealed the presence of a single polypeptide of 51 kDa. SDS-PAGE of CP3 reacted with 2-mercaptoethanol revealed the presence of two polypeptides of 31 and 18 kDa, indicating a dimer structure (alpha 1 beta 1) of the enzyme with disulfide-linked subunits. By using [1,3-3H]diisopropylfluorophosphate as an active-site labeling reagent, it was determined that the catalytic site resides on the small subunit of the carboxypeptidase. With N-carboben zoxy-L-phenylalanyl-L-leucine (N-CBZ-Phe-Leu) as the substrate, the Km, kcat, and Vmax values were 1.7 x 10(-4) M, 490 s-1, and 588 mumol of Leu released per min per mg of protein, respectively. CP3 was determined to be a serine protease, since its catalytic activity was blocked by the serine protease inhibitors diisopropylfluorophosphate, phenylmethylsulfonyl fluoride, and 3,4-dichloroi Socoumarin (DCI). The enzyme was strongly inhibited by the mercurial compound p-chloromercuribenzoate. The carboxypeptidase readily hydrolyzed peptides with aliphatic or aromatic side chains, whereas most of the peptides which contained glycine in the penultimate position did not serve as substrates for the enzyme. Although CP3 activity was undetectable in Mucor yeast cells, antisera revealed the presence of the enzyme in the yeast form of the fungus. The partial amino acid sequence of the carboxypeptidase was determined.

Amino Acid Sequence↗

Localization of functional projections from corpus callosum to cerebral cortex.

In 30 patients with intractable generalized epilepsy treated with cerebral commissurotomy, the corpus callosum was stimulated intraoperatively at a 1 cm interval with electric current, and evoked potentials (EPs) were recorded from different areas of the brain for determining the distribution patterns of functional projections from the corpus callosum to the cerebral cortex. The surface of the corpus callosum in man was 12 cm long and it was divided into 12 segments (1 cm each). Stimulation of the first segment resulted in EPs only in the frontal lobe. Stimulation of segments 2-4 produced EPs mainly in the anterior, middle and posterior frontal lobe, anterior and central temporal lobe, rarely in parietal and occipital lobes. Stimulation of segments 5-8 induced EPs mainly in the frontal and temporal lobes, but rarely in the parietal and occipital regions; on stimulation of segments 9-11, EPs occurred only in the parietal and occipital regions. These results were confirmed electrophysiologically in 5 cats and anatomically in 8 cats. These data provide an anatomical basis for selective cerebral commissurotomy in the treatment of intractable epilepsy. Hence in patients with a concentration of epileptic discharges in the frontal lobe, the 4 anterior segments of the corpus callosum should be incised. Epileptic discharges in the frontotemporal region indicates that the middle segments should be incised and epileptic activities originating from the parieto-occipital regions can be treated effectively by selective section of the posterior corpus callosum (segments 9-12).

Adolescent↗