Search PubMed⌕ Search

Biomedical subjects

T Liu

Publications and source records attributed to T Liu.

At least 253 records · Page 14Linked to original sources
PubMed1999

The relationship between expressions of N-myc and c-myc oncogenes in neuroblastoma: an in situ hybridization and immunocytochemical study.

N-myc gene amplification is the most characteristic feature of neuroblastoma. c-myc oncogene, another member of myc gene family, plays an important role in cell proliferation and differentiation. Both of them may contribute to tumorigenesis of neuroblastoma. In this study we use the in situ hybridization and immunocytochemical methods to test the frequencies of N-myc and c-myc expressions in 20 cases of human neuroblastoma at mRNA and protein levels. The positive rates of the expression of N-myc are 90% and 100% detected by in situ hybridization and immunocytochemical methods respectively. The positive rates of c-myc are 80% and 85% respectively. Sixty percent of the 20 specimens tested by in situ hybridization and 55% by immunocytochemistry show an inverse relationship between the expressions of these two oncogenes and this may indicate that there are different gene expression controlling mechanisms in different cases.

Adrenal Gland Neoplasms↗
PubMed1998

Identification of two regions in the cytoplasmic domain of CD44 through which PMA, calcium, and foskolin differentially regulate the binding of CD44 to hyaluronic acid.

CD44 contains two clustered basic residues of three arginines and three lysines in the membrane-proximal region of its cytoplasmic domain. These two clusters are conserved among different species and different splicing forms. The function of these two motifs is not known. We substituted either the three-arginine or the three-lysine motif with alanine (CD44.3R3A and CD44.3K3A) and established stable CD44 transfectants. The effects of these mutations on the binding of CD44 to one of its ligands, hyaluronic acid (HA), were studied. When stimulated with PMA, transfectants bearing CD44.3K3A and CD44.3R3A proteins have reduced HA-binding capacity. When stimulated with forskolin, an activator of cAMP-dependent PKC, CD44.3R3A transfectants were able to bind low but detectable level of fluorescent-conjugated HA (F-HA). In contrast, CD44.3K3A transfectants were unable to bind any F-HA. Elevation of intracellular calcium concentrations either by ionomycin or thapsigargin also induced binding of HA in CD44.3R3A but not in CD44. 3K3A transfectants. These results provide evidence that both the arginine and the lysine motifs are important in the binding of CD44 to high levels of HA when stimulated with PMA. In contrast, when transfectants were stimulated with either forskolin or a Ca2+ mobilizer to bind a low level of F-HA, the lysine cluster, but not the arginine cluster, is required. These two closely located basic clusters are, therefore, differentially involved in the binding of CD44 to HA, depending on the level of ligand binding and the nature of the stimulatory signals.

Arginine↗
PubMed1998

Mass spectrometric sequencing of site-specific carcinogen-modified oligodeoxyribonucleotides containing bulky benzo[a]pyrene diol epoxide-deoxyguanosyl adducts.

Site-specific carcinogen-modified oligonucleotides are often used in site-directed mutagenesis and other biological and biochemical studies of structure-function relationships. Postsynthetic analysis and confirmation of the sites of carcinogen binding in such oligonucleotides is an important step in the characterization of these site-specific carcinogen-DNA adducts. It is shown here that negative ion mode electrospray tandem mass spectrometry methods and collision-induced dissociation offer a rapid and convenient approach for the sequencing of products derived from the reaction of the carcinogenic and mutagenic metabolite of benzo[a]pyrene, the diol epoxide r7,t8-dihydroxy-t9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE), with the 11-mer oligonucleotide d(CATGCGGCCTAC). The site of reaction of anti-BPDE with either one of the three dG residues in this oligonucleotide can be accurately established by comparing the mass/charge ratios of the observed collision-induced dissociation fragments with calculated values.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1998

Mutagenic potential of stereoisomeric bay region (+)- and (-)-cis-anti-benzo[a]pyrene diol epoxide-N2-2'-deoxyguanosine adducts in Escherichia coli and simian kidney cells.

We have investigated the mutagenic potential of site-specifically positioned DNA adducts with (+)- and (-)-cis-anti stereochemistry derived from the binding of r7,t8-dihydroxy-t9,10-epoxy-7,8,9, 10-tetrahydrobenzo[a]pyrene (BPDE) to N2-2'-deoxyguanosine (G1 or G2) in the sequence context 5'TCCTCCTG1 G2CCTCTC. BPDE-modified oligodeoxynucleotides were ligated to a single-stranded DNA vector and replicated in Escherichia coli or simian kidney (COS7) cells. The presence of (+)- or (-)-cis adduct strongly reduced the yield of transformants in E. coli, and the yield was improved by the induction of SOS functions. Both adducts were mutagenic in E. coli and COS cells, generating primarily G --> T transversions. In E. coli, the (-)-cis adduct was more mutagenic than the (+)-cis adduct, while in COS cells, both adducts were equally mutagenic. These results were compared with those obtained with stereoisomeric (+)- and (-)-trans adducts [Moriya, M., et al. (1996) Biochemistry 35, 16646-16651). In E. coli, cis adducts, especially (-)-cis adducts, are consistently more mutagenic than the comparable trans adduct. In COS cells, trans adducts yield higher frequencies of mutations than the two cis adducts and, with the exception of the high-mutation frequency associated with the (+)-trans adduct at G2, relatively small differences in mutation frequencies are observed for the three other adducts. In E. coli, mutation frequency is a pronounced function of adduct stereochemistry and adduct position. These findings suggest that the fidelity of translesional synthesis across BPDE-dG adducts is strongly influenced by adduct stereochemistry, nucleotide sequence context, and the DNA replication complex.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1998

Mechanisms regulating the binding activity of CD44 to hyaluronic acid.

CD44 is a cell surface glycoprotein present on many cell types. Many CD44 isoforms have been identified. All CD44 isoforms utilize identical transmembrane and cytoplasmic domains. The hematopoietic form of CD44 (CD44H) is the major CD44 protein present on normal human lymphocytes and monocytes. One of the ligands for CD44 is hyaluronic acid (HA), a polymer consisting of repeat units of disaccharide; N-acetyl-D-glucosamine and N-acetyl-D-glucuronic acid. Since HA is present ubiquitously in extracellular matrix and in circulation, promiscuous binding of HA to CD44 may have undesirable affect. Similar to other adhesion molecules, binding of HA to cell surface CD44 requires regulation. In this review, we summarized our studies using a human lymphoma cell line, Jurkat. We found that binding of CD44+ Jurkat transfectants to HA requires cellular activation. Cellular activation induces the reorganization of the cytoskeleton proteins. Reorganization of cytoskeletal proteins results in clustering of CD44 on the cell surface. Clustering of CD44 on the cell surface is a prerequisite for the homodimerization of CD44. Our studies on Jurkat transfectants and results from other investigators suggest that interactions between CD44 and HA is a dynamic process and requires the participation of different cellular components; depending of the nature of the cell type and/or the nature of the activation signals.

Amino Acid Sequence↗
PubMed1998

[Detection of SMN gene deletions in spinal muscular atrophy].

OBJECTIVE: Survival motor neuron gene(SMN) and neuronal apoptosis inhibitory protein gene (NAIP) have been identified as the candidates of progressive spinal muscular atrophy (SMA)-determining genes. The aims of this study were to investigate the absence of SMN gene exon 7 in Chinese SMA patients, to confirm the relationship between the deletion of the SMN and SMA further, and to establish methods for gene diagnosis and prenatal diagnosis of SMA. METHODS: PCR-SSCP with silver staining method was used to detect the genomic DNA of 37 SMA patients and 30 normal individuals for deletions of SMN exon 7. RESULTS: Homozygous deletion of the SMN exon 7 was identified in 86.7%(13/15) of type I SMA patients and 86.4%(19/22) of type II patients. In the 88 controls (including parents of patients and normal individuals), homozygous absence of SMA exon 7 was only found in a mother of a patient. CONCLUSION: The data support that homozygous absence of SMN exon 7 is strongly associated with SMA. The percentage of homozygous deletions in this study is almost as high as that reported by other researchers. This method is useful, reliable and effective for gene diagnosis and prenatal diagnosis of SMA.

Cyclic AMP Response Element-Binding Protein↗
PubMed1998

Viscosity-adjustable block copolymer for DNA separation by capillary electrophoresis.

The viscosity-adjustable property of F127 block copolymer PEO99PPO69PEO99, PEO and PPO being poly(ethylene oxide) and poly(propylene oxide), respectively, was found to be useful for the development of automated capillary electrophoresis (CE). The polymer solution can form a gel-like structure with sieving ability and can also serve as a dynamic coating material, thereby effectively suppressing the electroosmotic flow induced by the ionization of the silanol group on the quartz capillary inner wall. When applied to CE as a separation medium, F127 block copolymer can provide the advantages of high separation resolution, easy injection and replacement of the triblock copolymer solution and convenient capillary column treatment. High reproducibility of DNA electrophoretic migration time in CE by replenishing F127 solution in acid-washed capillary tubings can be achieved. The relative standard deviation of the DNA migration time is less than 2%. In the investigation of F127 concentration and temperature effects on the performance of DNA separation in CE, we have found that the DNA electrophoretic migration behavior in the F127 gel-like solution cannot be described by any one of the existing models.

DNA↗
PubMed1998

Application of free-flow electrophoresis to the purification of trichosanthin from a crude product of acetone fractional precipitation.

The application of free-flow electrophoresis (FFE) to the purification of trichosanthin (TCS) from a crude product of acetone fractional precipitation was investigated. An electrophoresis technique, combining field step electrophoresis (FSE) and zone electrophoresis (ZE) to a one-step procedure, was optimized until a satisfactory purification factor (1.35), high resolution, and purity (>99%) were achieved. Testing several separation buffer systems revealed that a throughput of 14.2 mg/h can be obtained when the very basic TCS (pI 10.1) was dissolved and electrophoresed in a phosphate buffer system of pH 4. The purity of electrophoresed trichosanthin was proved by a variety of analytical methods, such as sodium dodecyl sulfate (SDS)-gel electrophoresis, capillary isoelectric focusing (CIEF), and sequencing of N- and C-termini. The high purity and large throughput achieved at low cost by using FFE indicates that this method can be employed for TCS purification.

Acetone↗
PubMed1998

The role for endothelin in mesangial proliferative glomerulonephritis.

To understand the effect of endothelin (ET) on the mesangium and matrix proliferation on the basis of hyperlipidemia, the model of mesangial proliferative glomerulonephritis (MSPGN) was established by intravenously giving calf serum albumin (250 mg/L) to the rabbits fed on feed containing 1% cholesterol for 4-8 weeks. Our results showed that in the 4th week of the experiment, the ET level in experiment group was significantly higher than that of normal controls (P < 0.01) and the pathological examination revealed development of MSPGN. In the 8th week of the experiment, the ET level was further elevated and pathological study demonstrated local glomerular sclerosis and infiltration of lymphocytes of interstitia. We are led to conclude that ET, as a newly identified hormone, is involved in the pathogenesis and development of MSPGN and acts as an important regulator in the proliferation of mesangium and accumulation of matrix of basal membrane.

Animals↗
PubMed1998

Differential effects of bone mineral content and bone area on vertebral strength in a swine model.

Since the biomechanical competence of a vertebral body may be closely related to the content and distribution of the bone mineral, we have evaluated the effects of projected vertebral bone area (BA) and bone mineral parameters [bone mineral content (BMC) or bone mineral density (BMD)] on their biomechanical competence. We used dual-energy X-ray absorptiometry (DXA) to assess the bone mineral parameters of 36 swine thoracic vertebrae (T1-T12) and 15 lumbar vertebrae (L1-L5) after removal of the posterior elements. The failure load, compressive stress, and the stored strain energy of these vertebral bodies were assessed by a uniaxial compressive test using an MTS 810 testing system. Multiple regression analysis showed a significantly negative effect of BA and significantly positive effect of BMC on the biomechanical competence (compressive stress, r2 = 0.67, P < 0.0001; failure load, r2 = 0.75, P < 0.0001). However, the stored strain energy was only related to the BMC (r2 = 0.35, P < 0.0001). The contributory effects of BMC and BA on the biomechanical competence were not equal. The effects of BMC was larger than BA in determining the failure load and stored strain energy, whereas the reverse was found for the compressive stress. Using the log-transformed parameters as the regressors resulted in similar results. These results suggested the differential effects of BA and BMC in determining the biomechanical competence of vertebral bodies. We recommend the use of both parameters instead of BMD alone for evaluation of the vertebral biomechanical competence.

Absorptiometry, Photon↗
PubMed1998

Preclinical pharmacology of the natural product anticancer agent 10-hydroxycamptothecin, an inhibitor of topoisomerase I.

PURPOSE: 10-Hydroxycamptothecin (HCPT) is an indole alkaloid isolated from a Chinese tree, Camptotheca acuminata, and has a wide spectrum of anticancer activity in vitro and in vivo mainly through inhibitory effects on topoisomerase I. HCPT has been shown to be more potent and less toxic than camptothecin and has recently undergone clinical trials. To determine how HCPT might be best used as an anticancer agent, preclinical studies of the pharmacokinetics, tissue distribution, metabolism and elimination of HCPT in rats were undertaken. METHODS: HCPT was administered to rats by i.v. bolus injection at doses of 1, 3, and 10 mg/kg body weight. HCPT (lactone and carboxylate) and its metabolites in plasma, urine, feces, and various tissues were quantitated by reversed-phase HPLC. Pharmacokinetic parameters were then estimated. RESULTS: Following i.v. administration at doses of 3 or 10 mg/kg, the plasma concentration-time profile for lactone HCPT could be best described by a three-compartment model, with terminal elimination half-lives of 140.4 and 428.6 min, respectively. A two-compartment model was used to fit the plasma concentration-time curve at 1 mg/kg, with a terminal elimination half-life of 30.5 min. Carboxylate HCPT had a longer half-life than the lactone form of HCPT. During the initial 6 h after dosing, urinary excretion was the major route of elimination, and fecal excretion became the major route of elimination thereafter. HCPT was widely distributed to various tissues including the enterohepatic system, kidney, and bone marrow. The lactone form of HCPT was detectable in various tissues examined up to 72 h after dosing at all the three test doses. HCPT glucuronides were present in plasma, urine, feces and various tissues. No significant toxicity was observed at doses of 1 or 3 mg/kg. Polyuria and hematuria were observed only during the initial 3 h after dosing at 10 mg/kg. CONCLUSIONS: Prolonged elimination of HCPT in vivo may have a significant impact on its therapeutic effects. HCPT is metabolized to its carboxylate form and glucuronides. Dose-dependent toxicity was observed with i.v. administration of HCPT. The results of this study should be useful in the design of future human trials with this anticancer drug.

Animals↗
PubMed1998

The effect of antibody against vascular endothelial growth factor on tumor growth and metastasis.

Vascular endothelial growth factor (VEGF), a very important in the process of tumor angiogenesis, was chosen as a target in a study to determine whether manipulation of angiogenesis with antibody against VEGF may interrupt tumor growth and metastasis. Anti-VEGF antibody was obtained from immunized rabbits, purified on an affinity column, and identified as neutralized antibody by Mile's assay. IVTA2MA891, a murine spontaneous breast cancer with a high rate of metastasis in lung in TA2 x 615 F1 mice, was chosen as an animal model in this study, because of the high expression of VEGF in the primary tumor as well as in the lung metastatic tumor. The anti-VEGF antibody could inhibit growth of S180 sarcoma in a dose-dependent manner, and the inhibition rate could reach 41.0% with a dose of 200 microg mouse(-1) day(-1). Anti-VEGF antibody could inhibit tumor growth by 76.2% in nude mice bearing human gastric cancer (MGC 803). When anti-VEGF antibody was combined with 131I-3H11, a murine monoclonal antibody conjugated with 131I, only one of five nude mice developed tumor and 84.0% more inhibition of tumor growth was obtained in comparison with treatment by 131I-3H11 alone. The growth of the primary tumor was inhibited by 44.0% and the number and size of the metastatic foci in the lungs were reduced by 73.0% and 83.7% respectively in the animal model, with a high rate of metastasis in lung. The anti-VEGF antibody may be potentially useful for clinical treatment of cancer and metastasis.

Animals↗
PubMed1998

Measurement of cardiac troponin T is an effective method for predicting complications among emergency department patients with chest pain.

STUDY OBJECTIVES: To determine the test performance characteristics of serum cardiac troponin T (cTnT) measurement for diagnosis of acute myocardial infarction (AMI), and to determine the ability of cTnT to stratify emergency department patients with chest pain into high- and low-risk groups for cardiac complications. METHODS: We conducted a prospective observational cohort study with convenience sampling in a tertiary care, urban ED. The study sample comprised 667 patients presenting to the ED with a complaint of chest pain or other symptoms suggesting acute ischemic coronary syndrome (AICS). Patients were assigned to different blood sampling protocols for cTnT therapy on the basis of their ECG at presentation: nondiagnostic for AMI at 0, 3, 6, 9, 12, and 24 hours after ED presentation; or ECG diagnostic for AMI at 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 12, 18, and 24 hours after ED presentation. RESULTS: Of 667 patients, 34 had AMI diagnosed within 24 hours of ED arrival. Using a .2 microgram/L discrimination level for cTnT, sensitivity for AMI within 24 hours of ED arrival was 97% (95% confidence interval, 91.4% to 99.9%), and specificity was 92% (89.8%-94.1%). When the effects of age, race, sex, and creatine kinase-MB isoenzyme subunit test results were controlled, a patient with cTnT of .2 microgram/L or greater was 3.5 (1.4 to 9.1) times more likely to have a cardiac complication within 60 days of ED arrival than a patient with a cTnT value below .2 microgram/L. CONCLUSION: Measurement of cTnT will accurately identify myocardial necrosis in patients presenting to the ED with possible AICS. Elevated cTnT values identify patients at increased risk of cardiac complications.

Adult↗
PubMed1998

The epitope stability of group 1 and group 2 allergens in mite extracts.

BACKGROUND: Commercial mite allergenic extracts sold in the US are prepared with whole body mites in buffer solutions containing 50% glycerol. Mite extracts were reported to contain large number of proteolytic enzymes and their structural integrity in aqueous solutions have not been evaluated. OBJECTIVE: To evaluate the epitope stabilities of Group 1 and 2 allergens in two commercial mite extracts used by Center for Biologics Evaluation and Research (CBER) as reference extracts, E5-Dp (Dermatophagoides pteronyssinus) and E5-Df (D. farinae). METHODS: Epitope stability was determined by using monoclonal antibodies in a Sandwich ELISA. Samples were stored at four different temperatures and the amounts of Der p 1, Der p 2, Der f 1, and Der f 2 were determined at different time intervals. The overall stability of mite extracts was evaluated by immunoblot and competition ELISA. RESULTS: The epitope stability of these allergens varies: Der f 1 was stable for at least 3 years and Der f 2 for 1 year when stored at 4 degrees C; Der p 1 and 2 were less stable. None of the Group 1 and 2 allergens remained intact when stored at 50 degrees C. Immunoblot and competition ELISA data also showed similar trend of degradation as compared with extracts stored at 4 degrees C for same length of time. CONCLUSION: With the exception of Der f 1, the amount of detectable epitopes in Group 1 and 2 allergens reduce rapidly after 1 year, especially at elevated temperatures. The changes in allergen composition were also observed by immunoblotting and in relative potency by ELISA competition assay. These findings are highly relavent to the users of CBER's mite extracts as standards.

Allergens↗
PubMed1998

Improving the fixation method for preimplantation genetic diagnosis by fluorescent in situ hybridization.

PURPOSE: Our purpose was to modify a fixation method using Tween-20 and HCl (TH) and to compare it with a protocol using methanol and acetic acid (MA) for the improvement of preimplantation genetic diagnosis by fluorescence in situ hybridization (FISH). METHODS: Single blastomeres were allocated to either the TH or the MA procedure. The two methods were compared to evaluate efficiency of fixation and the intensities of FISH signals. RESULTS: With the TH method, 123 (93.9%) of 131 blastomeres were fixed, while only 95 (78.5%) of 121 were fixed with MA. Average scores for the intensity of FISH signals were significantly stronger for TH than for MA (P < 0.05). There was also a significant difference in signal intensity scores between the two methods for type-3 nuclei. CONCLUSIONS: Our results indicate that not only are fewer nuclei lost during fixation but also stronger FISH signals can be obtained with the TH method. Thus, modified TH can improve the overall efficiency of preimplantation genetic diagnosis.

Acetic Acid↗
PubMed1998

The E protein of satellite phage P4 acts as an anti-repressor by binding to the C protein of helper phage P2.

Temperate phage P2 has the capacity to function as a helper for the defective, unrelated, satellite phage P4. In the absence of a helper, P4 can either lysogenize its host or establish itself as a plasmid. For lytic growth, P4 requires the structural genes, packaging and lysis functions of the helper. P4 can get access to the late genes of prophage P2 by derepression, which is mediated by the P4 E protein. E has been hypothesized to function as an anti-repressor. To locate possible epitopes interacting with E, an epitope display library was screened against E, and the most frequent sequence found had some identities to a region within P2 C. Using the yeast two-hybrid system, a clear activation of a reporter gene was found, strongly supporting an interaction between E and C. The P2 C repressor is believed to act as a dimer, which is confirmed in this work using in vivo dimerization studies. The E protein was also found to form dimers in vivo. The E protein only affects dimerization of C marginally, but the presence of E enhances multimeric forms of C. Furthermore, binding of the C protein to its operator is inhibited by E in vitro, indicating that the anti-repressor function of E is mediated by the formation of multimeric complexes of E and C that interfere with the binding of C to its operator.

Amino Acid Sequence↗