Biomedical subjects

X Tian

Publications and source records attributed to X Tian.

At least 19 recordsLinked to original sources

PubMed2003

99mTc-peptide-peptide nucleic acid probes for imaging oncogene mRNAs in tumours.

Imaging oncogene mRNA in tumours would provide a powerful tool for the early detection of occult malignant lesions. The goal was to prepare a chimera consisting of a dodecamer antisense peptide nucleic acid (PNA) specific for c-MYC oncogene overexpressed in human breast cancer cells and a chelating moiety that facilitates quantitative radiolabelling with 99mTc and evaluate it for hybridization and tissue distribution in laboratory animals. The pentapeptide chelator-PNA dodecamer specific for c-MYC mRNA was extended from a solid support by 9-fluorenylmethyloxycarbonyl (Fmoc) coupling. Similarly, a chelator-PNA chimera with four central mismatches was also prepared which served as a control. The chimeras were purified, characterized and evaluated for hybridization to c-MYC mRNA by fluorescent, real-time polymerase chain reaction (RT-PCR). The chimeras were labelled with 99mTc and their tissue distribution was examined in athymic nude mice bearing experimental human breast tumours. 99mTc radiolabelling was quantitative and presented a single peak in reversed phase liquid chromatography. Fluorescent real-time polymerase chain reactions using primer and fluorescent probe sets previously calculated for c-MYC mRNA demonstrated inhibition of reverse transcription by the c-MYC specific chimera as compared to that of the control. Tissue distribution studies of antisense and mismatch chimeras at 4 h and 24 h after administration displayed modest accumulation in the liver, and appreciable levels in tumours. These observations suggest that 99mTc-peptide-PNA probes might be useful for imaging gene expression in tumours, and the approach is worthy of further investigation.

Animals↗

PubMed2002

Therapeutically effective antibodies against amyloid-beta peptide target amyloid-beta residues 4-10 and inhibit cytotoxicity and fibrillogenesis.

Immunization of transgenic mouse models of Alzheimer disease using amyloid-beta peptide (Abeta) reduces both the Alzheimer disease-like neuropathology and the spatial memory impairments of these mice. However, a therapeutic trial of immunization with Abeta42 in humans was discontinued because a few patients developed significant meningo-encephalitic cellular inflammatory reactions. Here we show that beneficial effects in mice arise from antibodies selectively directed against residues 4-10 of Abeta42, and that these antibodies inhibit both Abeta fibrillogenesis and cytotoxicity without eliciting an inflammatory response. These findings provide the basis for improved immunization antigens as well as attempts to design small-molecule mimics as alternative therapies.

Alzheimer Vaccines↗

PubMed2002

Bipedal stance exercise and prostaglandin E2 (PGE2) and its synergistic effect in increasing bone mass and in lowering the PGE2 dose required to prevent ovariectomized-induced cancellous bone loss in aged rats.

Previous reports have shown that bone loss was partially prevented by bipedal stance "exercise" following ovariectomy (ovx), and it was well documented that prostaglandin E2 (PGE(2)) had an anabolic effect on the rat skeleton. The aim of this study was to determine whether lower doses of PGE(2) could prevent ovx-induced cancellous bone loss with the combination of bipedal stance exercise. Seventy-eight 10-month-old female Sprague-Dawley rats were either ovariectomized or sham-operated on day 0 and then treated with PGE(2) (0, 0.3, or 1 mg/kg per day) and/or housed in normal height cages (NC, 28 cm) or raised cages (RC, 33 cm) for 8 weeks. Bone histomorphometry was performed on the double-fluorescent-labeled proximal tibial metaphysis. In sham rats, 1 mg/kg PGE(2) + RC had synergistic effects in increasing trabecular bone area, width, and number by stimulating mineral apposition rate and bone formation rate. As expected, ovx induced cancellous bone loss, accompanied by elevated activation frequency. Without RC, PGE(2) monotherapy prevented ovx-induced bone loss at the 1 mg/kg per day dose, whereas this prevention effect was observed at the 0.3 mg/kg per day dose when combined with RC. Similar to their effects in sham rats, PGE(2) and RC had synergistic effects in augmenting cancellous bone mass and architecture and maintaining the elevated bone formation but depressing bone resorption and activation frequency. We conclude that bipedal stance exercise lowers the PGE(2) dose required to prevent ovx-induced cancellous bone loss in the proximal tibial metaphysis in aged rats.

Animals↗

PubMed2001

Basis for signaling specificity difference between Sos and Ras-GRF guanine nucleotide exchange factors.

Sos and Ras-GRF are two families of guanine nucleotide exchange factors that activate Ras proteins in cells. Sos proteins are ubiquitously expressed and are activated in response to cell-surface tyrosine kinase stimulation. In contrast, Ras-GRF proteins are expressed primarily in central nervous system neurons and are activated by calcium/calmodulin binding and by phosphorylation. Although both Sos1 and Ras-GRF1 activate the Ras proteins Ha-Ras, N-Ras, and Ki-Ras, only Ras-GRF1 also activates the functionally distinct R-Ras GTPase. In this study, we determined which amino acid sequences in these exchange factors and their target GTPases are responsible for this signaling specificity difference. Analysis of chimeras and individual amino acid exchanges between Sos1 and Ras-GRF1 revealed that the critical amino acids reside within an 11-amino acid segment of their catalytic domains between the second and third structurally conserved regions (amino acids (aa) 828-838 in Sos1 and 1057-1067 in Ras-GRF1) of Ras guanine nucleotide exchange factors. In Sos1, this segment is in helix B, which is known to interact with the switch 2 region of Ha-Ras. Interestingly, a similar analysis of Ha-Ras and R-Ras chimeras did not identify the switch 2 region of Ha-Ras as encoding specificity. Instead, we found a more distal protein segment, helix 3 (aa 91-103 in Ha-Ras and 117-129 in R-Ras), which interacts instead primarily with helix K (aa 1002-1016) of Sos1. These findings suggest that specificity derives from the fact that R-Ras-specific amino acids in the region analogous to Ha-Ras helix 3 prevent a functional interaction with Sos1 indirectly, possibly by preventing an appropriate association of its switch 2 region with helix B of Sos1. Although previous studies have shown that helix B of Sos1 and helix 3 of Ha-Ras are involved in promoting nucleotide exchange on Ras proteins, this study highlights the importance of these regions in establishing signaling specificity.

Amino Acid Sequence↗

PubMed2001

Cloning and analysis of mold-specific genes in the dimorphic fungus Histoplasma capsulatum.

A critical feature in the pathogenesis of the respiratory pathogen Histoplasma capsulatum is the conversion from the mold form (found in soil) to the yeast form in the lungs of the host. Little is known about the molecular biology of Histoplasma dimorphism. In particular, the possible roles of genes which are transcriptionally silent in yeast (i.e. mold-specific) have not been studied. We have produced a cDNA library highly enriched for mold-upregulated clones by fragmenting cDNA and removing yeast-specific and common sequences with a highly efficient enzyme degrading subtraction method. Screening of randomly selected clones identified cDNA fragments representing 16 different mold-upregulated genes. Because multiple cDNA fragments can be treated as alleles in a genetic screen, we were able to apply probability analysis to estimate the total number of mold-upregulated genes. We estimate that there are 27 upregulated genes; cDNA fragments of 16 have been isolated. Here we report the first isolation and analysis of cDNA from two mold-specific genes, MS8 (GenBank AF292398) and MS88 (GenBank AF357882). The MS8 transcript was very strongly expressed in mold but not detected on Northern blots with yeast RNA. The putative MS8 protein was predicted to be 21.3 kDa (203 aa), very rich in glutamine and glycine and had a calculated pI of 6.76. The MS88 transcript was weakly expressed in mold and not detected in yeast. The putative MS88 protein was predicted to be 22.5 kDa (219 aa) with a pI of 4.46. GenBank similarity searches revealed that the putative MS8 protein was similar to a glutamine-rich protein, of unknown function, from the fungus Colletotrichum gloeosporioides (GenBank U94186). No significant matches were found for the putative MS88 protein.

Amino Acid Sequence↗

PubMed2001

Separation and determination of podophyllum lignans by micellar electrokinetic chromatography.

A micellar electrokinetic chromatography method was established for the quantitative analysis of seven podophyllum lignans in Podophyllum emodi Wall. var. chinesis sprague. The optimum buffer system was 10 mM NaH2PO4-5 mM borate-100 mM sodium dodecylsulfate-30% isopropanol (pH 7.20). Voltage was 18 kV and detection at 214 nm. The second derivative chromatogram was used to determine a low-content component and those not fully separated from adjacent ones. The RSD values of migration times and peak areas were <2.2 and <5.5%, respectively. The effects of several CE parameters on the resolutions were studied systematically.

Chromatography, Micellar Electrokinetic Capillary↗

PubMed2001

Vascular endothelial growth factor: acting as an autocrine growth factor for human gastric adenocarcinoma cell MGC803.

Vascular endothelial growth factor (VEGF) is known to be a highly specific mitogen for endothelial cells through two high-affinity tyrosine kinase receptors, VEGFR-1 and VEGFR-2, which are almost specifically expressed in endothelial cells. However, recent findings showed that VEGF receptors may also expressed by nonendothelial cells, especially by tumor cells. To further understand the functional expression of VEGF receptors by nonendothelial cells, our preliminary screening detected the expression of VEGFR-2 in 115 different paraffin-embedded cancer specimens including 35 cases of bladder tumor, 30 cases of breast cancer, 25 cases of intestinal cancer, and 25 cases of lung cancer with immunohistochemistry. The results showed that VEGFR-2 was widely expressed in different tumor tissues. By reverse transcription PCR, NCI-H23, NCI-H460, MGC803, MDA-MB-231, 293, and MCF7 cells were evaluated for the mRNA expression of both VEGF and VEGFR-2. The data indicated that all these tumor cell lines expressed detectable amounts of VEGF mRNA, but only 293, MCF7, and MGC803 cells coexpressed VEGFR-2. Immunoblot analysis also demonstrated the expression of VEGFR-2 at protein level. We further demonstrate that exogenous rhVEGF(165) could stimulate cell growth in MGC803, a tumor cell line derived from gastric adenocarcinoma, in a dose- and time-dependent manner. Furthermore, the antibodies against rhVEGF(165) and VEGFR-2 could block rhVEGF(165)-mediated proliferation of MGC803 cells. These unexpected results provided direct evidence that VEGF may act as an autocrine growth factor to induce the proliferation of gastric adenocarcinoma cells as well as tumor angiogenic cells, thus suggesting a promising tumor therapeutic application based upon the VEGF system.

Adenocarcinoma↗

PubMed2001

Prenylation of target GTPases contributes to signaling specificity of Ras-guanine nucleotide exchange factors.

Ras-GRF1 and Ras-GRF2 constitute a family of calmodulin-regulated guanine-nucleotide exchange factors (GEFs) that activate Ras proteins. Here we show that whereas Ras-GRF1 activated both Ha-Ras and R-Ras in cells, Ras-GRF2 activated only Ha-Ras. The inability of Ras-GRF2 to activate R-Ras was the consequence of the GTPase being post-translationally modified, since Ras-GRF2 activated unprocessed R-Ras as effectively as unprocessed Ha-Ras when assays were performed either in vivo or in vitro. Moreover, Ras-GRF2 failed to activate fully processed R-Ras in vitro. The particular C-terminal lipid attached to the GTPases played an important role in determining signaling specificity, since R-Ras became more responsive to Ras-GRF2 when it was farnesylated instead of geranylgeranylated. Similarly, Ha-Ras became less responsive to Ras-GRF2 when it was geranylgeranylated instead of farnesylated. Analysis of chimeras between Ras-GRF1 and Ras-GRF2 demonstrated that a 30-amino acid segment embedded with their catalytic domains was responsible for recognizing the presence of different lipids on Ras proteins. These results indicate that the specific lipid moiety attached to GTPases can contribute to signaling specificity of Ras-GEFs.

Amino Acid Sequence↗

PubMed2001

The development of a potential single photon emission computed tomography (SPECT) imaging agent for the corticotropin-releasing hormone receptor type.

A high-affinity radioligand for CRHR1 has been prepared that can serve as a template for the development of SPECT imaging agents. The 5-chloro-N-cyclopropylmethyl-N-(2,6-dichloro-4-iodophenyl)-2-methyl-N-propylpyrimidine-4,6-diamine (6b, Ki = 14 nM), and the corresponding 4-bromophenyl analogue (6a, Ki = 21 nM), were synthesized in four steps from compound 3.

Animals↗

PubMed2001

Enmein type diterpenoids from Isodon japonica.

Two enmein type diterpenoids, taibaijaponicains A and B, were isolated from the ethanol extract of the leaves and branches of Isodon japonica. Their structures are designated as 6beta,11alpha-dihydroxy-16alpha-methoxymethyl-6,20-epoxy-6,7-seco-ent-kaur-15-one-1,7-olide and 3beta-acetoxy-6beta,11alpha-dihydroxy-16alpha-methoxymethyl-6,20-epoxy-6,7-seco-ent-kaur-15-one-1,7-olide, respectively, on the basis of detailed spectroscopic analyses.

Diterpenes↗

PubMed2001

Triterpenoid saponins from Clematis tangutica.

Two new triterpenoid saponins, tanguticoside A and B along with seven known saponins vitalboside B, alpha-hederin, saponin PK, beta-hederin, saponin PJ3, saponin PE, and ciwujianoside A were isolated from aerial part of Clematis tangutica. By chemical and spectral evidences methods, the structures of tanguticoside A and B were elucidated as 3-O-beta-D-glucopyranosylhederagenin 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside and 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosylhederagenin 28-O-alpha-D-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside, respectively.

Drugs, Chinese Herbal↗

PubMed2001

Regulation of Ras signaling specificity by protein kinase C.

Ras proteins have the capacity to bind to and activate at least three families of downstream target proteins: Raf kinases, phosphatidylinositol 3 (PI 3)-kinase, and Ral-specific guanine nucleotide exchange factors (Ral-GEFs). We have previously shown that the Ras/Ral-GEF and Ras/Raf pathways oppose each other upon nerve growth factor stimulation, with the former promoting proliferation and the latter promoting cell cycle arrest. Moreover, the pathways are not activated equally. While the Ras/Raf/Erk signaling pathway is induced for hours, the Ras/Ral-GEF/Ral signaling pathway is induced for only minutes. Here we show that this preferential down-regulation of Ral signaling is mediated, at least in part, by protein kinase C (PKC). In particular, we show that PKC activation by phorbol ester treatment of cells blocks growth factor-induced Ral activation while it enhances Erk activation. Moreover, suppression of growth factor-induced PKC activation enhances and prolongs Ral activation. PKC does not influence the basal activity of the Ral-GEF designated Ral-GDS but suppresses its activation by Ras. Interestingly, Ras binding to the C-terminal Ras binding domain of Ral-GDS is not affected by PKC activity. Instead, suppression of Ral-GDS activation occurs through the region N terminal to the catalytic domain, which becomes phosphorylated in response to phorbol ester treatment of cells. These findings identify a role for PKC in determining the specificity of Ras signaling by its ability to differentially modulate Ras effector protein activation.

Animals↗

PubMed2001

Magnetic tracker calibration for an augmented reality system for therapy.

Magnetic trackers are widely used position/orientation measurement devices in interactive computer systems. These trackers output six-degree-of-freedom position and orientation of their receiver coordinates with respect to their transmitter coordinates. It is a potential solution for real-time position/orientation measurement in an augmented reality system for therapy (ART). However, an obstruction to be overcome is that a magnetic tracker is vulnerable to measurement inaccuracy caused by magnetic field distortions (including ferric and non-ferric distortions) when it is used in metallic environments. The purpose of this paper is to discuss calibration methods used to compensate distortions produced by metallic objects which are moving during the measurement procedure, but fixed with a receiver. Past work on the calibration of magnetic trackers has concentrated on static distortions, which are produced by stationary metallic objects in the environment. Compared with static distortions, in this paper a relatively dynamic situation is considered where the distortion is not constant with time. However, the location of the metallic objects is reflected in the outputs of the receiver so that the distortion can be compensated. This type of distortions, which is called 'quasi-static distortion' in this paper, is common in an ART. Compensation of quasi-static distortion is more complicated than that of static distortion and has not been mentioned in past publications. Some new compensation methods for quasi-static distortion will be introduced here. The results demonstrate that the methods are capable of significantly reducing quasi-static distortions.

Calibration↗

PubMed2001

Variations in cardiac diastolic function in hypertensive patients with different left ventricular geometric patterns.

To evaluate the alteration of cardiac function in hypertensive patients with different left ventricular geometric patterns. Echocardiography was used to study left ventricular geometry and cardiac diastolic function in 117 cases of essential hypertension, with 45 normal cases as controls. Echocardiographic date were used to calculated the left ventricular mass index (LVMI) and relative wall thickness (RWT), which values in turn were used to divide the subjects into four groups. The left atrial dimension of the group, with the exception of these hypertensives who showed normal geometry, was larger than that of the control group. The damage of peak of E velocity, peak of A velocity, E/A and the slope between the E and F points (E to F slope) were greater than in hypertension than in the control group. The concentric hypertrophy group and eccentric hypertrophy group suffered more serious damage of left ventricular diastolic function than the concentric remodeling group, and damage of left ventricular diastolic function in the concentric remodeling group was greater than that in the normal geometry group. The degree of cardiac diastolic function damage differed among patients with different left ventricular geometric patterns, when the cardiac structure was changed, the degree of cardiac diastolic function damage increased.

Diastole↗

PubMed2001

Resistant potato selections contain leptine and inhibit development of the Colorado potato beetle (Coleoptera: Chrysomelidae).

We recently described a new source of host-plant resistance to the Colorado potato beetle, Leptinotarsa decemlineata (Say), in a tetraploid potato (Solanum tuberosum L.) selection, ND2858-1. This genotype, and selected backcross progeny, had little damage while check cultivars were defoliated in open-choice field assays. To further characterize the observed deterrence, we determined foliar glycoalkaloids and conducted no-choice assays with ND2858-1 backcross progeny genotypes (ND4382-n). Development of neonate L. decemlineata in detached leaf assays on resistant progeny genotypes was delayed and larval weight gain after 4 d was inhibited by 75% relative to larval development and weight gain on susceptible genotypes. Inhibition of larval development in detached leaf assays with the selected progeny genotypes was equivalent to that of high-leptine genotypes of S. chacoense Bitter. Foliar glycoalkaloids of resistant genotypes included low levels of leptines I and II. The unlikely nature of this cross and the presence of leptine in this and resistant progeny selections cast doubt on the recorded pedigree. Molecular analyses were conducted by restriction fragment-length polymorphism and amplified fragment-length polymorphisms. Both methods established a high degree of relatedness to S. tuberososum and S. chacoense but not to S. fendleri. We conclude that ND2858-1 did not originate from a cross with S. fendleri, but is likely derived from S. chacoense. Oviposition and larval survival were reduced when adult L. decemlineata were placed in cages with resistant genotypes; an effect that was enhanced by inclusion of Perillus bioculatus F. Therefore, the nonpreference previously observed in open-choice field defoliation assays is also associated with antibiotic effects on L. decemlineata. The resistance may be caused by leptines, but is greater than would be expected by the leptine content. This source of host plant resistance could be a cost-effective management strategy, especially if combined with other resistance mechanisms or compatible control measures to delay development of resistance in the target insects.

Animals↗

PubMed2001

[Magnetic stimulation: theory and advance].

Magnetic stimulation is a noninvasive diagnostic and therapeutic technique. The basic theory of magnetic stimulation is presented briefly. Its advances are elaborated in three aspects of mechanism, instrumentation and applications. The problems in focality, location, safety and its trend are discussed.

Humans↗

PubMed2001

Disposable 1-day Acuvue contact lenses for the delivery of lomefloxacin to rabbits' eyes.

PURPOSE: To investigate the ability of a disposable soft contact lens (1-Day Acuvue) to deliver lomefloxacin, a fluoroquinolones antibiotic, into the cornea and aqueous humor of rabbits. METHODS: Experiments were conducted on adult albino rabbits. 1-Day Acuvue lenses were immersed for 1 hour in commercially-available lomefloxacin eye solution and then placed on the cornea. After 0.5, 2, 4,6, and 8 hours, the animals were killed and the lenses were removed and placed into a 20 mL saline bath. Corneal tissue and aqueous humor samples were also obtained at these times. The release of lomefloxacin from the lenses was calculated by measuring the amount of drug remaining in the lenses after removal from the rabbit eyes. The concentration of lomefloxacin in the cornea and anterior chamber following the wearing of lomefloxacin-loaded lenses was compared with the concentrations following frequent-drop therapy (one drop of lomefloxacin hourly for 8 hours). RESULTS: In vivo, lomefloxacin was released from 1-Day Acuvue lenses gradually over 8 hours. The cornea and aqueous humor levels in the eyes following the wear of lomefloxacin-loaded lenses were significantly higher than those achieved by frequent-drop therapy. The drug levels in the cornea and aqueous humor generally remained above the 90% minimal inhibitory concentration for 8 hours in the 1-Day Acuvue lens group. CONCLUSIONS: 1-Day Acuvue contact lenses soaked in 0.3% lomefloxacin can release sufficient amounts of lomefloxacin and in this study produced higher levels in both the cornea and aqueous humor than that achieved by frequent-drop therapy for up to 8 hours. We conclude that 1-Day Acuvue contact lens can be used as a drug delivery system for lomefloxacin.

Animals↗

PubMed2001

Studies on the uptake and release of fluoroquinolones by disposable contact lenses.

PURPOSE: To evaluate the ability of three types of disposable contact lenses to take up and release three kinds of fluoroquinolones in vitro. METHODS: We evaluated three FDA-approved disposable soft contact lenses (1-Day Acuvue, Medalist, and 14UV) for their ability to absorb and release three kinds of fluoroquinolones. Contact lenses were presoaked in fluoroquinolones for 1, 4, and 24 hours, and the uptake was determined by measuring the concentration of fluoroquinolones in the three types of disposable soft contact lenses by high pressure liquid chromatography (HPLC). After uptake, the lenses were placed in fresh saline baths, and release rates from the lenses were determined by measuring the concentration of fluoroquinolones in the saline baths by HPLC. RESULTS: The disposable soft contact lens with the highest uptake of fluoroquinolones was the 1-Day Acuvue. After presoaking, drug concentrations in the 1-Day Acuvue and Medalist were higher than their theoretical saturation concentrations (i.e., the concentration of the fluoroquinolone eye drops [0.3%]). The release rates from the 1-Day Acuvue and Medalist lenses were slower than for the 14 UV lens. CONCLUSIONS: These results indicate that among the lenses tested, the most practical drug delivery system is the 1-Day Acuvue disposable soft contact lens.

Anti-Infective Agents↗