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Biomedical subjects

X Liu

Publications and source records attributed to X Liu.

At least 361 records · Page 20Linked to original sources

BioProspector: discovering conserved DNA motifs in upstream regulatory regions of co-expressed genes.

The development of genome sequencing and DNA microarray analysis of gene expression gives rise to the demand for data-mining tools. BioProspector, a C program using a Gibbs sampling strategy, examines the upstream region of genes in the same gene expression pattern group and looks for regulatory sequence motifs. BioProspector uses zero to third-order Markov background models whose parameters are either given by the user or estimated from a specified sequence file. The significance of each motif found is judged based on a motif score distribution estimated by a Monte Carlo method. In addition, BioProspector modifies the motif model used in the earlier Gibbs samplers to allow for the modeling of gapped motifs and motifs with palindromic patterns. All these modifications greatly improve the performance of the program. Although testing and development are still in progress, the program has shown preliminary success in finding the binding motifs for Saccharomyces cerevisiae RAP1, Bacillus subtilis RNA polymerase, and Escherichia coli CRP. We are currently working on combining BioProspector with a clustering program to explore gene expression networks and regulatory mechanisms.

Bacillus subtilis↗

[Development of three-dimensional stereo viewer for high-resolution data].

In order to visualize high-resolution three-dimensional (3D) data as a stereogram, a real-time volume-rendering system using a hardware graphic board and conventional PC was developed. A 256(3) data set could be visualized at a redrawing rate of 12 Hz, and a 512(3) data set at a rate of 2.5 Hz. It was demonstrated that stereogram visualization using volume graphic hardware architecture potentially enables rapid examination of high-resolution 3D data by changing visualization parameters such as level, window, transfer function for opacity, and color map or coordinate direction.

Computer Graphics↗

Identification of key residues responsible for enzymatic and platelet-aggregation-inhibiting activities of acidic phospholipase A2S from Agkistrodon halys Pallas.

Site-directed mutagenesis was used to probe the structural and functional relationship of acidic phospholipase A2 from Agkistrodon halys Pallas. The mutants are AP-E6R (E6R), AP-D115K (D115K), AP-6R115K (E6R, D115K), AP-Y118M (Y118M), and AP-W119T (W119T). All mutants were inserted into a bacterial expression vector and effectively expressed in E. coli RR1. The purified recombinant enzymes were used to assay for enzymatic and inhibiting platelet aggregation activities. The enzymatic activities of AP-D115K, AP-Y118M and AP-W119T are close to that of denatured-refolded acidic phospholipase A2, while the enzymatic activities of AP-E6R, AP-6R1 15K are lower than that of denatured-refolded acidic phospholipase A2 (AP-WT). In these five mutants, AP-Y118M showed strongest inhibiting effect on platelet aggregation, which is the same as that of AP-WT, AP-W119T showed only modest activity and AP-E6R, AP-D115K, AP-6R115K showed little activity. To study the structural and functional relationships among these five mutants, molecular modeling of these five mutants was done. The roles of various amino acid residues in the enzymatic activity and pharmacological activity of acidic phospholipase A2 are discussed.

Adult↗

[Studies on neonatal screening, clinical and gene analysis for tetrahydrobiopterin deficiency in Southern Chinese].

OBJECTIVE: To find out the incidence of tetrahydrobiopterin deficiency(BH4D) among patients with hyperphenylalaninemia in Southern Chinese and evaluate the clinical outcome and gene mutations of tetrahydrobiopterin deficient patients. METHODS: Analyses of urinary neopterin(N) and biopterin(B) were done in 87 patients with hyperphenylalaninemia by high-performance liquid chromatography. The patients with BH4 deficiency and their parents were asked to undergo the gene mutation analysis and the patients were treated and followed up. RESULTS: Eleven cases of which the urinary N/B ratio was higher than 38 and B% lower than 5% were diagnosed as BH4 deficiency caused by 6-pyruvoyl-tetrahydropterin synthase(PTPS) deficiency. The incidence of BH4 deficiency among patients with hyperphenylalaninemia is 12% in Southern Chinese. PTPS gene mutations (P87S, N52S, D96N and G144R) were detected from 5 PTPS deficient families. The G144R mutation is a new mutation. The five PTPS-deficient patients were treated with synthetic BH4, neurotransmitter precursors L-dopa and 5-hydroxytryptophan. They had satisfactory physical and mental development after treatment, and 4 of them scored their IQ 70-80. CONCLUSION: The screening for BH4 deficiency should be carried out in all patients with hyperphenylalaninemia in order to minimize the misdiagnoses.

Biopterins↗

[Dietary intake of antioxidant vitamins and personal demographic characteristics of Guangdong residents].

The study aim at revealing the associations between the intake of dietary antioxidant vitamins and personal characteristics on age, sex, education, working status and household gross income etc. The intake of antioxidant vitamins in a population of Guangdong Province was studied. A total of 418 males and 503 females, aged 25-84 year were interviewed, and questionnaires on socio-demography and on a 12-month food intake frequency (FFQ) were completed in 1995. The daily average intake of antioxidant vitamins and energy was higher in male than in female. Older subjects consumed lesser antioxidant vitamins. The vitamin E intake of higher education persons as well as high family income females (> 15001RMB) was higher. Individual business owners and farmers consumed lesser antioxidant vitamins than the other counterparts. The highest vitamin E intake was found in the currently unemployed men and retired women. BMI was positively associated with the intake of antioxidant vitamins excepted for the obesity group. The result suggests that the consumption of antioxidant vitamins varies with characteristics of socio-demographic status. Health promotion programs, such as suggestions on eating more fruits and vegetables should be targeted at the elderly, the less educated, lower income families and rural community.

Adult↗

[Determination of protocatechualdehyde in yu xue bi granula].

The content of Protocatechualdehyde in YuXueBi granula was determined by HPLC. The chromatographic column was packed with ODS-2 (particle size 5 microns). The mobile phase was MeOH-H2O-H3PO4(17:83:0.003) adjusted to pH 3 with N(C2H5)3. The detection wavelength was 280 nm. The velocity of flow was 1.0 ml/min. The average recovery rate was 96.14%, RSD was 1.16%. The method was simple and stable.

Benzaldehydes↗

Colon epithelial cell death in 2,4,6-trinitrobenzenesulfonic acid-induced colitis is associated with increased inducible nitric-oxide synthase expression and peroxynitrite production.

Peroxynitrite, derived from the reaction of nitric oxide (NO(.)) with superoxide (O(2)), is a potent nitrating and oxidizing agent that can induce apoptosis in a variety of different cell types. In the present study, we investigated the possible role of peroxynitrite as a mediator of colon epithelial cell death in rat colitis. Rat colon inflammation was induced by intracolonic administration of 2,4,6-trinitrobenzenesulfonic acid (TNBS) and rats were sacrificed 24 h after TNBS administration. Expression of inducible nitric-oxide synthase (iNOS) was detected by reverse transcription-polymerase chain reaction and immunohistochemistry. The enzymatic activities of Ca(2+)-independent iNOS and Ca(2+)-dependent constitutive nitric-oxide synthase were determined biochemically. Evidence of peroxynitrite-mediated cell injury was detected by immunostaining of nitrotyrosine. Apoptosis was examined by in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay and DNA gel electrophoresis. To evaluate the specific contribution of peroxynitrite to the observed cell injury, a selective iNOS inhibitor, L-N(G)-[1-iminoethyl]lysine (L-NIL), was administered after TNBS induction. Morphological examination and analysis of TUNEL/cytokeratin double immunofluorescence revealed significant apoptosis in mucosal epithelial cells. Nitrotyrosine was colocalized with TUNEL, strongly demonstrating the association of peroxynitrite with the apoptotic death of colon epithelial cells. The administration of L-NIL reduced iNOS activity in 24-h lesions by 92% and also significantly attenuated both nitrotyrosine staining and apoptotic cell counts in the colon epithelium. These results strongly suggest that local elevated level of peroxynitrite produced from increased iNOS expression and activity is a major contributor to colon epithelial apoptosis during colon inflammation.

Animals↗

[Analysis on genetic polymorphism of 13 STR loci on chromosomes 11 and 19 in Chinese Hans].

OBJECTIVE: To analyze the genetic polymorphism of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 in Chinese Hans. METHODS: Polymerase chain reaction-single strand length polymorphism(PCR-SSLP) was used to genotype 100 randomly selected individuals of the Han nationality at 8 STR loci(D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359) on chromosome 11 and 5 STR loci(D19S247, D19S714, D19S433, D19S246, D19S254) on chromosome 19. RESULTS: Eight alleles and 26 genotypes, 9 alleles and 15 genotypes, 6 alleles and 16 genotypes, 12 alleles and 40 genotypes, 6 alleles and 19 genotypes, 12 alleles and 48 genotypes, 8 alleles and 20 genotypes, 7 alleles and 13 genotypes were observed at D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359. The heterozygosities for the 8 STR loci were 87%, 68%, 73%, 92%, 71%, 86%, 75% and 71%, respectively. Ten alleles and 19 genotypes, 10 alleles and 26 genotypes, 10 alleles and 24 genotypes, 11 alleles and 29 genotypes, 8 alleles and 18 genotypes were observed at D19S247, D19S714, D19S433, D19S246, D19S254. The heterozygosities for the 5 STR loci were 63%, 82% 72%, 81% 74%, respectively. CONCLUSION: The distribution of allele frequencies of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 was consistent with the Hardy-Weinberg equilibrium and the highly genetic polymorphism was observed in Chinese Han population.

China↗

[Association analysis of attempted suicide and 5-HT2A receptor gene].

OBJECTIVE: To examine the relationship of attempted suicide with A-1438G and T102C polymorphisms of 5-HT2A receptor gene. METHODS: Genotypes and allele frequencies of A-1438G and T102C polymorphisms of 5-HT2A receptor gene in 149 attempted suicides and 190 normal controls were examined by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). RESULTS: No differences in genotypes and allele frequencies of A-1438G and T102C polymorphisms of 5-HT2A receptor gene were observed between attempted suicides and the controls. However, an association between male attempted suicides and genotype(GG) of A-1438G polymorphism was found. CONCLUSION: These results suggested that liability to suicide behavior might be associated with A-1438G polymorpyhism of 5-HT2A receptor gene in male subjects.

Adolescent↗

Delayed dark-adaptation and lipofuscin accumulation in abcr+/- mice: implications for involvement of ABCR in age-related macular degeneration.

PURPOSE: To examine the ocular phenotype in mice heterozygous for a null mutation in the abcr gene. METHODS: Retinas and retinal pigment epithelia (RPE) were prepared from wild-type, abcr+/-, and abcr-/- mice. Fresh tissues were homogenized and analyzed by normal phase high-performance liquid chromatography (HPLC) for the presence of retinoids and phospholipids. In another study, fixed tissues were sectioned and analyzed by light and electron microscopy. Finally, anesthetized mice were studied by electroretinography (ERG) at different times after exposure to strong light. RESULTS: A2E, the major fluorophore of lipofuscin, and its precursors, A2PE-H(2) and A2PE, were approximately fourfold more abundant in 8-month-old abcr+/- than in the wild-type retina and RPE. The levels of these substances in abcr+/- mice were approximately 40% those in abcr-/- mice. Lipofuscin pigment-granules were also visible in abcr+/- RPE cells by electron microscopy. Accumulation of A2PE-H(2) and A2E in abcr+/- retina and RPE, respectively, was strongly dependent on light exposure. Heterozygous mutants also exhibited delayed recovery of rod sensitivity by ERG. This delay was correlated with elevated levels of all-trans-retinaldehyde (all-trans-RAL) in retina after a photobleach and was not caused by a reduction in quantum-catch due to depletion of 11-cis-retinaldehyde (11-cis-RAL). CONCLUSIONS: Partial loss of the ABCR or rim protein is sufficient to cause a phenotype in mice similar to recessive Stargardt's disease (STGD) and age-related macular degeneration (AMD) in humans. These data are consistent with the suggestion that the STGD carrier-state may predispose to the development of AMD.

ATP-Binding Cassette Transporters↗

Vascular endothelial growth factor promotes proliferation and migration of cavernous smooth muscle cells.

PURPOSE: We investigated whether cavernous smooth muscle cells secrete vascular endothelial growth factor (VEGF), how they respond to VEGF and how age affects their ability to secrete and respond to VEGF. MATERIALS AND METHODS: Corpus cavernous tissues were isolated from rats of various ages and grown as monolayer cell cultures. Smooth muscle identity was determined by immunofluorescence staining. Secreted VEGF was measured with a VEGF enzyme-linked immunosorbent assay kit. Cell proliferation was assayed with a cell titer kit. Cell migration was measured with Transwell chambers. VEGF receptor (VEGFR) messenger RNA and proteins were identified by reverse transcription-polymerase chain reaction and immunoblotting, respectively. RESULTS: Cavernous smooth muscle cells from young (age 1 week) and old (age 28 months) rats secreted the least amounts of VEGF and those from 16-week-old rats secreted the most. VEGF stimulated the growth of cavernous smooth muscle cells at an optimal concentration of 12.5 ng./ml. At this concentration cavernous smooth muscle cells from 11-week-old and 28-month-old rats showed the highest and lowest growth rates, respectively. The optimal VEGF concentration for cavernous smooth muscle cell migration was 10 ng./ml. At this concentration cells from 4-week-old and 28-month-old rats showed the highest and lowest migratory rates, respectively. The expression of VEGFR-1 appeared to parallel the VEGF regulated growth rates of these cells. VEGFR-2 was undetectable in cavernous smooth muscle cells. CONCLUSIONS: Cavernous smooth muscle cells from rats of different ages secreted different amounts of VEGF and showed different abilities to respond to VEGF. Cavernous smooth muscle cells responded to VEGF in terms of cell proliferation and migration. Cavernous smooth muscle cells from rats of different ages expressed different levels of VEGFR-1 but did not express VEGFR-2.

Aging↗

[Effect of different cultured conditions on endothelial cell and its resistance to the fluid imposed shear stress--a comparative study].

This study was intended to enhance the resistance of cultured endothelial cells (EC) to fluid shear stress and clarify the possible mechanism. A modified parallel plate flow apparatus was used to compare the strength between the grafts and the endothelial cells under shear stress condition and static condition. The dynamic change of cytoskeletal actin filaments and the effects of different adhesive proteins coated on the strength of EC adhesion to the glass were studied. The results showed that the number of cells retention of the shear stress-conditioned group was significantly larger than that of the static group. Precoated fibronectin and laminin significantly promoted the adherence of the EC cultured in a steady environment but seemed to have no effect on the EC cultured under flow condition. The results suggest that the vascular endothelial cell cytoskeleton, stress fiber and the shape of EC reorganize in response to long term fluid-imposed shear stress, and the endothelial cells become tightly adherent to the grafts.

Actins↗

[Effect of flow shear stress on the expression of adhesion molecules of endothelial cells].

Adherence of various leukocytes, including monocyte, to vascular endothelial cells may play an important role in the development of atherosclerosis. In vivo, the hemodynamic shear forces have a critical effect on the surface expression of adhesion proteins. In order to elucidate the effect of flow shear stress on the expression of adhesion molecules of endothelial cells, we investigated the effect of flow shear stress (2.23-6.08 dyne/cm2) on the expression of adhesive molecules, intercellular adhesion molecule(ICAM-1), vascular cell adhesion molecule (VCAM-1), and E-selectin on cultured human umbilical vein endothelial cells(HUVECs). The expression of adhesion molecules on the surface of HUVECs induced by shear stress was analyzed using flow cytometry. The results showed that compared with stationary control, the surface expression of ICAM-1 was significantly increased (P < 0.05) on HUVECs after exposure to different shear stress (2.23, 4.20, 6.08 dyne/cm2), and it was found to be in close relationship with the shearing time (r = 0.992, 0.997, 0.997; P < 0.05). After exposure to shear(2.23 dyne/cm2), VCAM-1 expression was significant increased(P < 0.05), and it was positively correlated with the shearing time(r = 0.930; P < 0.05), while VCAM-1 expression dropped down to basal level(P < 0.05) after it was sheared at 4.20 or 6.08 dyne/cm2, and the magnitude of the reduction of VCAM-1 expression was negatively correlated with the shearing time (r = -0.975, -0.989; P < 0.05). E-selectin expression was less sensitive to shear stress, especially at the lower magnitudes of shear. These results indicate that the increase of ICAM-1 and VCAM-1 expression in endothelial cells induced by low shear stress may play a prominent role in the development of both inflammation and atherosclerosis.

Cells, Cultured↗

MYC messenger RNA expression predicts survival outcome in childhood primitive neuroectodermal tumor/medulloblastoma.

PURPOSE AND EXPERIMENTAL DESIGN: Cerebellar primitive neuroectodermal tumors/medulloblastomas (PNET/MB) are the most common malignant brain tumors in childhood. To identify PNET/MB biological prognostic factors that define a patient group with a sufficiently good prognosis to permit a reduction in treatment intensity, we determined the expression levels of MYC mRNA in fresh frozen tumor samples from 26 PNET/MB patients using semiquantitative reverse transcription-PCR. RESULTS: MYC mRNA expression levels in primary PNET/MB showed a wide range with a 22-fold difference between the highest and lowest values and did not correlate with MYC gene amplification. MYC mRNA expression was an independent significant prognostic factor for progression-free survival outcome and was more predictive than standard clinical factors. The combination of low MYC mRNA expression and high TrkC mRNA expression identified a good outcome group of PNET/MB patients (n = 7) with 100% progression-free survival after a median follow-up time of 55 months (range, 15-91 months). Three of these seven good outcome patients survived without radiotherapy. CONCLUSIONS: Low MYC mRNA expression is a powerful independent predictor of favorable clinical outcome in PNET/MB. Assessment of MYC mRNA levels is feasible and may be incorporated in prospective PNET/MB clinical trials to aid in treatment planning for patients with PNET/MB on confirmation of our results in larger studies.

Cerebellar Neoplasms↗

[Effects of tumor necrosis factor alpha on the expression of connective tissue growth factor in hepatic stellate cells].

OBJECTIVE: To investigate the effect of tumor necrosis factor alpha (TNF-alpha) on the expression of connective tissue growth factor (CTGF) in rat hepatic stellate cells (HSC) in vitro. METHODS: HSCs were prepared from SD rats by in situ perfusion and single-step Nycodenz gradient, and were treated with TNF-alpha and TGF-beta 1. The expression of CTGF in HSC was assessed using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Both TNF-alpha and TGF-beta 1 could stimulate the expression of CTGF mRNA in HSC. The expression of CTGF was not found in HSC after treatment with TNF-alpha(10microgram/L) for 6 hours, but was observed after TNF-alpha treatment for 24 and 48 hours. TGF-beta1 could induce the mRNA expression for CTGF just after 6 hours of treatment at 1microgram/L concentration. CONCLUSIONS: TNF-alpha is able to induce the expression of CTGF in HSC, and this may be one of the mechanisms for TNF-alpha involving in the liver fibrosis.

Animals↗

[The influence of ecological factors on simultaneous nitrification and denitrification in MBR].

The influence of dissolved oxygen(DO), C/N and pH on simultaneous nitrification and denitrification in the membrane bioreactor was studied. The experimental results showed that only under condition that each ecological factor was maintained relative steady, simultaneous nitrification and denitrification might proceed smoothly. When DO, C/N and pH of feed water was 1 mg/L, 30 and 7.2, the removal rate of COD, NH4+, and TN were 96%, 95% and 92% respectively.

Bioreactors↗