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Biomedical subjects

Bin Liu

Publications and source records attributed to Bin Liu.

At least 37 records · Page 2Linked to original sources

Quantitative intracellular molecular profiling using a one-dimensional flow system.

We report on the development of one-dimensional microfluidic bead arrays for rapid and quantitative molecular profiling of human cancer cells. This new bioanalytical platform integrates the rapid binding kinetics of suspension bead carriers, the multiplexing and encoding capabilities of gene/protein chips, and the liquid handling advantages of microfluidic devices. Using antibody-conjugated beads in a two-site "sandwich" format, we demonstrate that the proteomic contents of as few as 56 human lung epithelial cancer cells can be determined with high sensitivity and specificity. The results indicate that each cell contains approximately 6 x 10(5) copies of the tumor suppressor protein P53. We have further examined the expression changes of P53, c-Myc, and beta-Actin as a function of anticancer drug treatment and have validated these changes by using Western blotting. This ability to quantitatively analyze normal and diseased cells raises new possibilities in studying cancer heterogeneity and circulating tumor cells.

Animals↗

Effects of estrogen and related agents upon methamphetamine-induced neurotoxicity within an impaired nigrostriatal dopaminergic system of ovariectomized mice.

Estrogen increases methamphetamine (MA)-induced neurotoxicity within the impaired nigrostriatal dopaminergic (NSDA) system of ovariectomized female mice, as defined by enhanced striatal dopamine (DA) depletion. In this study we compared the effects of a lower dose of estradiol benzoate (EB, 1 microg) with related agents--tamoxifen (TMX, 12.5 microg), testosterone (5 microg) and dehydroepiandrosterone (DHEA, 3 mg) in this paradigm. In experiment 1, ovariectomized mice received an initial treatment with MA. At 1 week after MA, mice were treated with EB, TMX, testosterone, DHEA or oil vehicle and 24 h later a second MA treatment. Striatal DA and 3,4-dihydroxyphenylacetic acid (DOPAC) concentrations in the MA-treated groups were decreased compared to the non-MA-treated control. Neither EB nor any of the other agents tested showed enhanced neurodegenerative or neuroprotective effects against a second MA invasion. To verify that estrogen was capable of showing a neuroprotective effect under a condition of two administrations of MA, in experiment 2, EB was administered either once or twice prior to each of the two MA treatments. EB treatment prior to the first MA invasion or first and second MA protected the NSDA system against DA and DOPAC depletion. These results imply that a lower dose of EB, TMX, testosterone and DHEA cannot exert neurodegenerative or neuroprotective effects in the impaired NSDA model. However, EB administered prior to the introduction of neurotoxicity can protect the NSDA system. This study may provide an understanding of the variations in results on the effects of estrogen upon the NSDA neurodegenerative disorder, Parkinson's disease.

3,4-Dihydroxyphenylacetic Acid↗

Differentiation of rat bone marrow stem cells in liver after partial hepatectomy.

AIM: To investigate the differentiation of rat bone marrow stem cells in liver after partial hepatectomy. METHODS: Bone marrow cells were collected from the tibia of rat with partial hepatectomy, the medial and left hepatic lobes were excised. The bone marrow stem cells (Thy(+)CD3(-)CD45RA(-) cells) were enriched from the bone marrow cells by depleting red cells and fluorescence-activated cell sorting. The sorted bone marrow stem cells were labeled by PKH26-GL in vitro and autotransplanted by portal vein injection. After 2 wk, the transplanted bone marrow stem cells in liver were examined by the immunohistochemistry of albumin (hepatocyte-specific marker). RESULTS: The bone marrow stem cells (Thy(+)CD3(-)CD45RA(-) cells) accounted for 2.8% of bone marrow cells without red cells. The labeling rate of 10 microM PKH26-GL on sorted bone marrow stem cells was about 95%. There were sporadic PKH26-GL-labeled cells among hepatocytes in liver tissue section, and some of the cells expressed albumin. CONCLUSION: Rat bone marrow stem cells can differentiate into hepatocytes in regenerative environment and may participate in liver regeneration after partial hepatectomy.

Animals↗

Duplication of photoinduced azo polymer surface-relief gratings through a soft lithographic approach.

In this work, a soft lithographic approach has been developed to duplicate photoinduced surface-relief-gratings (SRGs) of azo polymer films to generate the surface pattern replicas composed of different materials on various substrates. For this purpose, thin films of an epoxy-based azo polymer (BP-AZ-CA) were prepared by spin-coating, and SRGs with different structures were inscribed by exposing the films to interference patterns of Ar(+) laser beams at modest intensity (150 mW/cm(2)). Using the azo polymer films as masters, stamps of poly(dimethylsiloxane) (PDMS) were prepared by replica molding. The PDMS stamps were then used to transfer the solutions of poly(3-hexylthiophene) (P3HT), multiwalled carbon nanotube (MWNT), and BP-AZ-CA to different substrates by contact printing. Through this process, surface pattern replicas made of the functional materials were obtained. The pattern formation and quality depended on the factors such as the solution concentration, contacting time in the printing process, and printing pressure. Under the proper conditions, the printed patterns showed the same grating periods as the masters and the same relief depths as the stamps (replicas of the masters). This approach, showing some attractive characteristics such as the easiness of master preparation and the versatility of soft fabrication processes, can be applied to the fabrications of optical functional surfaces, sensors, and photonic devices.

Journal Article↗

Sex differences in methamphetamine-evoked striatal dopamine of mice are reversed by nomifensine.

Male mice show more severe striatal dopamine depletions to the psychostimulant, methamphetamine (MA). To gain some understanding for this sex difference, we examined MA-evoked dopamine (DA) responses from superfused striatal tissue fragments of male and female mice under conditions of a dopamine transporter which was either unaltered (Experiment 1) or inhibited, with use of the drug, nomifensine (Experiment 2). In Experiment 1, MA-evoked DA was significantly greater in male versus female mice. In Experiment 2, diminished, albeit statistically significant, DA responses to MA infusion in the presence of nomifensine were obtained from striatal tissue of female, but not male, mice. In Experiment 3, potassium-evoked DA responses and sex differences were abolished in the presence of nomifensine. These data demonstrate a clear sex difference in DA responses to MA. Interestingly, under conditions where dopamine transporter function is inhibited, MA retains its ability to evoke DA. However, this capacity was only observed within striatal tissue fragments of female mice and not under conditions of potassium-evoked DA. These results indicate an additional component for the bases of sex differences in nigrostriatal dopaminergic function in health and in disease. In particular, the present findings have important implications in suggesting an alternative, non-traditional, mechanism for MA effects and indicate that such a function is limited to females.

Animals↗

Epidermal growth factor receptor activation: an upstream signal for transition of quiescent astrocytes into reactive astrocytes after neural injury.

Modulating the behaviors of reactive astrocytes is a potential therapeutic strategy for neurodegenerative diseases. We found that upregulation and activation of the epidermal growth factor receptor (EGFR) occur in astrocytes after different injuries in optic nerves in vivo. Activation of EGFR regulates genes and cellular processes representing most major markers of reactive astrocytes and genes related with glaucomatous optic neuropathy and other neural disorders. These results suggest that activation of EGFR is a common, regulatory pathway that triggers quiescent astrocytes into reactive astrocytes in response to neural injuries in the optic nerve, and perhaps other parts of the CNS. Targeting EGFR activation using an EGFR tyrosine kinase inhibitor prevents the loss of retinal ganglion cells in a model of glaucomatous optic neuropathy. Because these inhibitors are currently used clinically, our results present an approach to reactive astrocytes as a potential new target for the treatment of neurodegenerations.

Animals↗

Two novel bacteriophages of thermophilic bacteria isolated from deep-sea hydrothermal fields.

Bacteriophages of thermophiles are of great interest due to their important roles in many biogeochemical and ecological processes. However, no virion has been isolated from deep-sea thermophilic bacteria to date. In this investigation, two lytic bacteriophages (termed Bacillus virus W1 and Geobacillus virus E1) of thermophilic bacteria were purified from deep-sea hydrothermal fields in the Pacific for the first time. Bacillus virus W1 (BVW1) obtained from Bacillus sp. w13, had a long tail (300nm in length and 15 nm in width) and a hexagonal head (70 nm in diameter). Another virus, Geobacillus virus E1 (GVE1) from Geobacillus sp. E26323, was a typical Siphoviridae phage with a hexagonal head (130 nm in diameter) and a tail (180 nm in length and 30 nm in width). The two phages contained double-stranded genomic DNAs. The genomic DNA sizes of BVW1 and GVE1 were estimated to be about 18 and 41 kb, respectively. Based on SDS-PAGE of purified virions, six major proteins were revealed for each of the two phages. The findings in our study will be very helpful to realize the effect of virus on thermophiles as well as the communities in deep-sea hydrothermal fields.

Bacillaceae↗

Characterizing of tissue microstructure with single-detector polarization-sensitive optical coherence tomography.

Assessing tissue birefringence with imaging modality polarization-sensitive optical coherence tomography (PS-OCT) could improve the characterization of in vivo tissue pathology. Among the birefringent components, collagen may provide invaluable clinical information because of its alteration in disorders ranging from myocardial infarction to arthritis. But the features required of clinical imaging modality in these areas usually include the ability to assess the parameter of interest rapidly and without extensive data analysis, the characteristics that single-detector PS-OCT demonstrates. But beyond detecting organized collagen, which has been previously demonstrated and confirmed with the appropriate histological techniques, additional information can potentially be gained with PS-OCT, including collagen type, form versus intrinsic birefringence, the collagen angle, and the presence of multiple birefringence materials. In part I, we apply the simple but powerful fast-Fourier transform (FFT) to both PS-OCT mathematical modeling and in vitro bovine meniscus for improved PS-OCT data analysis. The FFT analysis yields, in a rapid, straightforward, and easily interpreted manner, information on the presence of multiple birefringent materials, distinguishing the true anatomical structure from patterns in image resulting from alterations in the polarization state and identifying the tissue/phantom optical axes. Therefore the use of the FFT analysis of PS-OCT data provides information on tissue composition beyond identifying the presence of organized collagen in real time and directly from the image without extensive mathematical manipulation or data analysis. In part II, Helistat phantoms (collagen type I) are analyzed with the ultimate goal of improved tissue characterization. This study, along with the data in part I, advance the insights gained from PS-OCT images beyond simply determining the presence or absence of birefringence.

Algorithms↗

[Inhibiting cerebral glioma growth with continuous low-dose chemotherapy and cyclooxygenase-2 inhibitor in nude mice].

OBJECTIVE: To observe the changes in glioma growth characteristic and apoptosis of tumor cells after single handed continuous low-dose chemotherapy, cyclooxygenase-2 inhibitor treatment alone and the combination of the two treatments. METHODS: The U251MG cells in exponential phase of growth were made into 10(7)/mL cell suspension in free-serum 1640 and stored in 37 degrees C incubator. The survival rate of cells was above 95%. The U251MG cells were implanted into the right parietooccipital lobe of the 4-week old nude mouse with a 5 micro liter micro amount sample injector. The number of injected U251MG cells was 5 x 10(4) for a mouse. Twenty days after the model making, the nude mice were treated with elemene and indometacin respectively and the combination of them, twice a week. The mice were divided into four groups. Group I was treated with indometacin alone, group II elemene alone, group III low-dose elemene plus indometacin, Group IV was used as controls, including tumor control and blank control. The animals were killed on the 40th and 50th day after implantation by breaking cervical vertebra. The fixed brain was made into 3 microm slices by paraffin section. The slices were carried out with HE staining and immunohistochemical staining of glial fibrillary acidic protein(GFAP), cell proliferation-associated antigen(Ki-67), cyclooxygenase-2(COX-2), CD34, programmed cell death 5(PDCD5) and terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL). RESULTS: The proliferation of glioma cells was predominant in the tumor control mouse brain. Several immature blood vessels were observed in the tumor implanted for 40 days. The white matter was infiltrated by bulk glioma cells along with capillary vessel clusters in the mouse brain implanted for 50 days. In groups with combination treatment of the two drugs, 40 days after the implantation, several apoptosis cells and glioma cells were observed in tumor where the positive signal for GFAP was showed; and 50 days after the implantation, lots of apoptosis cells were observed in tumor cell implantation area where the negative signal for GFAP and positive signal for PDCD5 was showed. The volume of tumor was (29.8+/-39.1) mm(3) 40 days after the implantation, and (78.4+/-125.9) mm(3) 50 days after the implantation. There was no statistically significant difference in tumor volume among groups(P=0.11). CONCLUSION: The combination of two treatments could merely prolong the survival time of the nude mouse model, without the effect of eliminating the tumor completely.

Animals↗

Leptin enhances in vitro secretion of IgG antiplatelet antibodies by splenocytes and peripheral blood mononuclear cells from patients with chronic idiopathic thrombocytopenic purpura.

Chronic idiopathic thrombocytopenic purpura (ITP) is an organ-specific autoimmune disease characterized by the production of antibodies against antigens on the membranes of platelets, resulting in enhanced destruction of the platelets by macrophages. Leptin, a cytokine-like hormone, plays a role in the pathogenesis of several autoimmune diseases. In this study, we observed significantly increased plasma leptin levels combined with decreased soluble leptin receptor (sOB-R) levels in 18 chronic ITP patients compared with 14 controls. We also demonstrated significantly elevated mRNA expression of long form leptin receptor (Ob-Rb) on peripheral blood mononuclear cells (PBMCs) from chronic ITP patients. Treatment with recombinant leptin or serum with high leptin levels enhanced in vitro secretion of IgG antiplatelet antibodies by splenocytes and PBMCs from patients with chronic ITP. After depletion of CD4(+) T cells from splenocytes, leptin lost this function. Further studies showed that leptin could increase platelet reactive T cells. These findings suggest that leptin may be involved in the pathogenesis of chronic ITP and might offer a potential target for the treatment of this disease.

Adult↗

A novel nuclear-localized protein with special adenylate kinase properties from Caenorhabditis elegans.

The adrenal gland protein AD-004 like protein (ADLP) from Caenorhabditis elegans was cloned and expressed in Escherichia coli. Enzyme assays showed that ADLP has special adenylate kinase (AK) properties, with ATP and dATP as the preferred phosphate donors. In contrast to all other AK isoforms, AMP and dAMP were the preferred substrates of ADLP; CMP, TMP and shikimate acid were also good substrates. Subcellular localization studies showed a predominant nuclear localization for this protein, which is different from AK1-AK5, but similar to that of human AK6. These results suggest that ADLP is more likely a member of the AK6 family. Furthermore, RNAi experiments targeting ADLP were conducted and showed that RNAi treatment resulted in the suppression of worm growth.

Adenylate Kinase↗

Synthesis, characterization and properties of chromium(III) complex [Cr(SA)(en)2]Cl.2H2O.

The reaction of chromium(III) chloride, salicylic acid (SA) and ethylenediamine (en) led to the formation of chromium complex [Cr(SA)(en)(2)]Clx2H(2)O(1). The crystal structure belongs to monoclinic system with the space group P2(1), R(1)=0.0358. In this compound, Cr(III) atom is six-coordinated in octahedral coordination geometry by one phenolic hydroxyl oxygen, one carboxylate oxygen from the salicylic acid and four nitrogen atoms from two ethylenediamine molecules, respectively. The transfer manners of Cr(III) from the title compound to the low-molecular-mass chelator, ethylenediamine-N,N,N',N'-tetraacetic acid (EDTA) and the iron-binding protein apoovotransferrin (apoOTf) were followed by a combination of UV-visible (UV-Vis) and fluorescence spectra in 0.01M Hepes at pH 7.4. The results show that Cr(III) can be transferred from the complex to apoovotransferrin with the retention of the salicylate acted as a synergistic anion.

Chlorides↗

Structural and molecular characterization of Shigella boydii type 16 O antigen.

Shigella is a well-known human pathogen causing dysentery and their typing is solely based on the O antigens. We investigated the chemical structure and gene cluster of Shigella boydii type 16 O antigen. As judged by sugar and methylation analyses along with NMR spectroscopy data, the O antigen has an O-acetylated branched pentasaccharide repeating O unit, which consists of two D-mannose residues (D-Man), one residue each of d-glucuronic acid (D-GlcA), N-acetylglucosamine (D-GlcNAc) and D-galactose (D-Gal), and the structure of the O unit was established. The O antigen gene cluster of S. boydii type 16 was identified and shown to contain putative genes for the synthesis of GDP-D-Man, genes encoding sugar transferases, O unit flippase (Wzx) and O antigen polymerase (Wzy) as expected. The function of the wzy gene was characterized by mutation test. Genes specific to S. boydii type 16 O antigen gene cluster were identified by screening 186 Escherichia coli and Shigella type strains, and can be used to develop PCR assays for detection of type 16 strains.

Base Sequence↗

Structural analyses on intermediates in serine protease catalysis.

Although the subject of many studies, detailed structural information on aspects of the catalytic cycle of serine proteases is lacking. Crystallographic analyses were performed in which an acyl-enzyme complex, formed from elastase and a peptide, was reacted with a series of nucleophilic dipeptides. Multiple analyses led to electron density maps consistent with the formation of a tetrahedral species. In certain cases, apparent peptide bond formation at the active site was observed, and the electron density maps suggested production of a cis-amide rather than a trans-amide. Evidence for a cis-amide configuration was also observed in the noncovalent complex between elastase and an alpha1-antitrypsin-derived tetrapeptide. Although there are caveats on the relevance of the crystallographic data to solution catalysis, the results enable detailed proposals for the pathway of the acylation step to be made. At least in some cases, it is proposed that the alcohol of Ser-195 may preferentially attack the carbonyl of the cis-amide form of the substrate, in a stereoelectronically favored manner, to give a tetrahedral oxyanion intermediate, which undergoes N-inversion and/or C-N bond rotation to enable protonation of the leaving group nitrogen. The mechanistic proposals may have consequences for protease inhibition, in particular for the design of high energy intermediate analogues.

Amides↗

Pharmacological characterization of endocannabinoid transport and fatty acid amide hydrolase inhibitors.

: 1. The mechanism of anandamide uptake and disposal has been an issue of considerable debate in the cannabinoid field. Several compounds have been reported to inhibit anandamide uptake or fatty acid amide hydrolase (FAAH; the primary catabolic enzyme of anandamide) activity with varying degrees of potency and selectivity. We recently reported the first evidence of a binding site involved in the uptake of endocannabinoids that is independent from FAAH. There are no direct comparisons of purported selective inhibitory compounds in common assay conditions measuring anandamide uptake, FAAH activity and binding activity. 2. A subset of compounds reported in the literature were tested in our laboratory under common assay conditions to measure their ability to (a) inhibit [(14)C]-anandamide uptake in cells containing (RBL-2H3) or cells lacking (HeLa) FAAH, (b) inhibit purified FAAH hydrolytic activity, and (c) inhibit binding to a putative binding site involved in endocannabinoid transport in both RBL and HeLa cell membranes. 3. Under these conditions, nearly all compounds tested inhibited (a) uptake of [(14)C]-anandamide, (b) enzyme activity in purified FAAH preparations, and (c) radioligand binding of [(3)H]-LY2183240 in RBL and HeLa plasma membrane preparations. General rank order potency was preserved within the three assays. However, concentration response curves were right-shifted for functional [(14)C]-anandamide uptake in HeLa (FAAH(-/-)) cells. 4. A more direct comparison of multiple inhibitors could be made in these three assay systems performed in the same laboratory, revealing more information about the selectivity of these compounds and the relationship between the putative endocannabinoid transport protein and FAAH. At least two separate proteins appear to be involved in uptake and degradation of anandamide. The most potent inhibitory compounds were right-shifted when transport was measured in HeLa (FAAH(-/-)) cells suggesting a requirement for a direct interaction with the FAAH protein to maintain high affinity binding of anandamide or inhibitors to the putative anandamide transport protein.

Amidohydrolases↗

PDGF-BB enhances monocyte chemoattractant protein-1 mRNA stability in smooth muscle cells by downregulating ribonuclease activity.

Platelet-derived growth factor (PDGF) has protean manifestations, including the regulation of growth and migration, in many cell types. We have previously reported that PDGF-BB induces the accumulation of monocyte chemoattractant protein (MCP)-1 mRNA in smooth muscle cells (SMC), in large part due to an increase in mRNA stability. To elucidate the mechanism by which PDGF-BB stabilizes MCP-1 mRNA, we have employed in vitro RNA gel mobility shift and decay assays. Cytoplasmic extracts from PDGF-BB-treated SMC increased the half-life of in vitro transcribed MCP-1 mRNA from approximately 45 min to >2 h. PDGF-BB-inhibitable degradation was not dependent on specific regions of the MCP-1 mRNA and was equally effective on a variety of in vitro transcribed mRNAs. Angiotensin II had a similar effect on MCP-1 mRNA stability, whereas tumor necrosis factor-alpha and basic fibroblast growth factor did not. The PDGF-BB-inhibitable RNAse activity was active at pH 6.6 and heat stable, but was sensitive to proteinase K. Extracts from PDGF-BB- or angiotensin II-treated cells inhibited the RNAse activity of control extracts, suggesting that the effect of PDGF-BB and angiotensin II are due to activation of a soluble inhibitor of the RNAse. The effect of PDGF-BB was blocked by inhibitors of tyrosine phosphorylation, but not by inhibitors of phosphatidylinositol 3-kinase or mitogen-activated protein kinases. These studies provide new insights into the mechanisms by which PDGF-BB enhances mRNA accumulation.

Angiotensin II↗