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Biomedical subjects

Hao Zhou

Publications and source records attributed to Hao Zhou.

At least 37 records · Page 2Linked to original sources
PubMed2005

Calculation of the electric polarizability of a charged spherical dielectric particle by the theory of colloidal electrokinetics.

Dielectrophoresis (DEP) is increasingly being explored as a means to manipulate or separate colloidal particles. The direction and strength of the DEP force depend strongly on the induced dipole strength, K, of a polarized particle, and predictions of DEP forces require carefully computed values for K. In this paper, we present the calculation of the dipole strength using the full electrokinetic theory of Mangelsdorf and White for both static and oscillating electric fields. The effects of particle zeta potential, radius, Debye length and electrolyte composition on the magnitude and sign of Re(K) are discussed. The full theory model is compared with the extended Maxwell-Wagner (EMW) model and the results show that the EMW model can fail to predict the full Re(K) variation with frequency, even predicting Re(K) with the incorrect sign depending on system parameters. A program for the dipole strength calculation shown in this paper is available from the authors.

Colloids↗
PubMed2005

Lateral separation of colloids or cells by dielectrophoresis augmented by AC electroosmosis.

Colloidal particles and biological cells are patterned and separated laterally adjacent to a micropatterned electrode array by applying AC electric fields that are principally oriented normally to the electrode array. This is demonstrated for yeast cells, red blood cells, and colloidal polystyrene particles of different sizes and zeta-potentials. The separation mechanism is observed experimentally to depend on the applied field frequency and voltage. At high frequencies, particles position themselves in a manner that is consistent with dielectrophoresis, while at low frequencies, the positioning is explained in terms of a strong coupling between gravity, the vertical component of the dielectrophoretic force, and the Stokes drag on particles induced by AC electroosmotic flow. Compared to high frequency dielectrophoretic separations, the low frequency separations are faster and require lower applied voltages. Furthermore, the AC electroosmosis coupling with dielectrophoresis may enable cell separations that are not feasible based on dielectrophoresis alone.

Cell Membrane↗
PubMed2005

Metal-responsive transcription factor (MTF-1) handles both extremes, copper load and copper starvation, by activating different genes.

From insects to mammals, metallothionein genes are induced in response to heavy metal load by the transcription factor MTF-1, which binds to short DNA sequence motifs, termed metal response elements (MREs). Here we describe a novel and seemingly paradoxical role for MTF-1 in Drosophila in that it also mediates transcriptional activation of Ctr1B, a copper importer, upon copper depletion. Activation depends on the same type of MRE motifs in the upstream region of the Ctr1B gene as are normally required for metal induction. Thus, a single transcription factor, MTF-1, plays a direct role in both copper detoxification and acquisition by inducing the expression of metallothioneins and of a copper importer, respectively.

Animals↗
PubMed2005

Fibrillogenesis of apomyoglobin facilitated by aggregation sequence of yeast Sup35 in various regions.

To examine the effect of aggregation sequence QGGYQQQYNP from yeast Sup35 on fibril formation of sperm whale apomyoglobin (apoMb), we constructed several mutants via substitution. Urea-induced unfolding of apoMb confirms that the substitution of the aggregation sequence does not significantly affect the stability of the mutants compared to wild type (WT) at pH 4.2. Under this condition, however, despite the difference in rate most apoMb mutants form fibrils more readily than WT with distinct morphology. These results suggest that the aggregation sequence facilitates fibril assembly of apoMb at acidic pH in vitro and this facilitation depends on the regions replaced.

Amino Acid Sequence↗
PubMed2005

Contributions of disulfide bonds in a nested pattern to the structure, stability, and biological functions of endostatin.

Endostatin can inhibit the proliferation and migration of endothelial cells. It contains two pairs of disulfide bonds in a nested pattern. We constructed three mutants, C33A/C173A, C135A/C165A, and all-Ala, to evaluate the contributions of individual disulfide bonds to the structure, stability, and biological functions of endostatin. Both tryptophan emission fluorescence spectrum and 1H nuclear magnetic resonance spectrum show that C135A/C165A and all-Ala, the two mutants lacking disulfide bond Cys135-Cys165, lost nearly their entire tertiary structure. Although C33A/C173A appears to retain some native-like structures, it is less stable and has a higher helical content, which confirms our earlier hypothesis that the polypeptide backbone of endostatin has a high helical propensity. C135A/C165A and all-Ala mutants lost most of their inhibitory activities both on the migration and proliferation of human microvascular endothelial cells, whereas C33A/C173A is partially active. The mutants without disulfide bond Cys135-Cys165 can hardly be internalized and localized to cytoskeleton and nucleus in the cell, which probably contributes to their loss of inhibition on the migration and proliferation of endothelial cells. Our studies provide a structural basis for the two disulfide bonds on the biological functions of endostatin.

Amino Acid Sequence↗
PubMed2005

Dynamic distribution of Ser-10 phosphorylated histone H3 in cytoplasm of MCF-7 and CHO cells during mitosis.

The dynamic distribution of phosphorylated Histone H3 on Ser10 (phospho-H3) in cells was investigated to determine its function during mitosis. Human breast adenocarcinoma cells MCF-7, and Chinese hamster cells CHO were analyzed by indirect immunofluorescence staining with an antibody against phospho-H3. We found that the phosphorylation begins at early prophase, and spreads throughout the chromosomes at late prophase. At metaphase, most of the phospho-H3 aggregates at the end of the condensed entity of chromosomes at equatorial plate. During anaphase and telophase, the fluorescent signal of phospho-H3 is detached from chromosomes into cytoplasm. At early anaphase, phospho-H3 shows ladder bands between two sets of separated chromosome, and forms "sandwich-like structure" when the chromosomes condensed. With the cleavage progressing, the "ladders" of the histone contract into a bigger bright dot. Then the histone aggregates and some of compacted microtubules in the midbody region are composed into a "bar-like" complex to separate daughter cells. The daughter cells seal their plasma membrane along with the ends of the "bar", inside which locates microtubules and modified histones, to finish the cytokinesis and keep the "bar complex" out of the cells. The specific distribution and kinetics of phospho-H3 in cytoplasm suggest that the modified histones may take part in the formation of midbody and play a crucial role in cytokinesis.

Animals↗
PubMed2005

[The clinical study of the epithelial flap of painless LASEK and Epi-LASIK].

OBJECTIVE: To protect epithelial flap and improve the rate of painless LASEK and Epi-LASIK surgery. METHODS: 285 eyes of 285 cases of LASEK, 39 eyes of 39 cases of Epi-LASIK were involved in the study. Laser machines used in the study included Mel 80, lasersight LSX, NIDEK EC5000, VISX 4star, AOV (66 vision) and 217Z. The Epi-LASIK hinge position was routinely at 11-12 o'clock. The application time of alcohol for LASEK was 10-20 seconds. The hinge was routinely at 12-1 o'clock by the way of epithelial-CCC. RESULTS: Postoperative pain: 269 cases (91.23%) in LASEK and 36 cases (92.31%) in Epi-LASIK were quiet (patients did not complain pain and open the eyes in a nature way), There was no significant difference in the ration of postoperative pain between two groups. LASEK: CLs were taken off in 10 cases (3.51%) in 7-11 days. Epi-LASIK: CLs were taken off in 2 cases (5.13%) in 7-10 days. The main reason of delayed taking of the CLs was irregular edema of epithelium. CONCLUSIONS: The LASEK and Epi-LASIK are able to be a painless procedure. LASEK/Epi-LASIK can get quiet situation postoperatively with quickly recovery of effective vision by LASIK. Delayed taking off CLs is safe and effective to protect the survival epithelium in case of the epithelial flap delayed healing.

Adolescent↗
PubMed2004

Low stability of nucleocapsid protein in SARS virus.

The nucleocapsid protein (N protein) is one of the major virion structural proteins of a newly identified coronavirus, which has been confirmed as the causative agent of severe acute respiratory syndrome (SARS). The major function of N protein is to assemble the RNA of coronavirus. In the present study, the gene encoding the N protein was cloned and the protein was expressed, purified, and refolded as shown by (1)H NMR measurement. The maximal Trp emission wavelength occurs near 331 nm, suggesting substantial burial of Trp residues. Circular dichroism measurements indicate that N protein contains little alpha-helical structure. Acid titration shows that N protein begins to unfold near pH 5 and is fully denatured near pH 2.7, and the acid unfolding process is reversible. The physical and chemical properties of N protein indicate that its stability is low. N protein is denatured reversibly at pH 7.4 either by urea (with C(m) of 2.77 M and m value of 2.74 kcal mol(-1) M(-1)) or GdmCl (with C(m) of 1.46 M and m value of 4.50 kcal mol(-1) M(-1)). In the heat-induced denaturation in phosphate-buffered saline buffer, N-protein starts to unfold at 35 degrees C and is completely denatured at 55 degrees C, where SARS virus was also reported to be inactivated. We propose that the low stability of N protein may be critical for the stability and function of SARS virus.

Cations, Monovalent↗
PubMed2004

Acid-induced unfolding mechanism of recombinant human endostatin.

Endostatin is a potent angiogenesis inhibitor. The structure of endostatin is unique in that its secondary structure is mainly irregular loops and beta-sheets and contains only a small fraction of alpha-helices with two pairs of disulfide bonds in a nested pattern. We choose human endostatin as a model system to study the folding mechanism of this kind. Nuclear magnetic resonance (NMR), tryptophan emission fluorescence, and circular dichroism (CD) were used to monitor the unfolding process of endostatin upon acid titration. Urea-induced unfolding was used to measure the stability of endostatin under different conditions. Our results show that endostatin is very acid-resistant; some native structure still remains even at pH 2 as evidenced by (1)H NMR. Trifluoroethanol (TFE) destabilizes native endostatin, while it makes endostatin even more acid-resistant in the low pH region. Stability measurement of endostatin suggests that endostatin is still in native structure at pH 3.5 despite the decreased stability. Acid-induced unfolding of endostatin is reversible, although it requires a long time to reach equilibrium below pH 3. Surprisingly, the alpha-helical content of endostatin is increased when it is unfolded at pH 1.6, and the alpha-helical content of the polypeptide chain of unfolded endostatin increases linearly with TFE concentration in the range of 0-30%. This observation indicates that the polypeptide chain of unfolded endostatin has an intrinsic alpha-helical propensity. Our discoveries may provide clues for refolding endostatin more efficiently. The acid-resistance property of endostatin may have biological significance in that it cannot be easily digested by proteases in an acidic environment such as in a lysosome in the cell.

Acetic Acid↗
PubMed2004

Regiospecific, enantiospecific total synthesis of the 12-alkoxy-substituted indole alkaloids, (+)-12-methoxy-Na-methylvellosimine, (+)-12-methoxyaffinisine, and (-)-fuchsiaefoline.

[reaction: see text] The enantiospecific synthesis of 7-methoxy-d-tryptophan was completed by combination of the Larock heteroannulation process with a Schöllkopf-based chiral auxiliary in good yield. This ester was then employed in the first total synthesis of (+)-12-methoxy-Na-methylvellosimine, (+)-12-methoxyaffinisine, and (-)-fuchsiaefoline in regiospecific, stereospecific fashion in excellent overall yield. The asymmetric Pictet-Spengler reaction and enolate-driven palladium-catalyzed cross coupling processes served as key steps.

Journal Article↗
PubMed2004

Arsenic trioxide inhibits proliferation in K562 cells by changing cell cycle and survivin expression.

To study the mechanisms involved in the inhibition of chronic myeloid leukemic cells (K562) proliferation induced by arsenic trioxide (As2O3) and to explore the potential role of Survivin, an inhibitor of apoptosis protein, in the regulation of As2O3 induced cell apoptosis, K562 cells were cultured with As2O3 of different concentrations. Cells were collected for proliferation analysis by MTT assay. Cell cycle distribution and cell apoptosis were analyzed by flow cytometry. Expression of Survivin protein and mRNA were detected by flow cytometry and RT-PCR, respectively. Our results showed that As2O3(2-10 micromol/L) inhibited K562 cells growth effectively, but it did not induce cells apoptosis significantly. The percentage of K562 cells at G2/M phase increased in proportion to As2O3 concentrations, and the expression of Survivin mRNA and content of Survivin protein was up-regulated accordingly. It is concluded that As2O3 inhibited K562 cells growth by inducing cell cycle arrest mainly at G2/M phase. Over-expression of Survivin gene and protein might be one of the possible mechanisms contributing to K562 cells' resistance to As2O3-induced apoptosis.

Antigens, Neoplasm↗
PubMed2004

Renal potassium handling in healthy and sick newborns.

Growing infants must maintain a state of positive K+ balance, a task accomplished, in large part, by the kidney. The distal nephron is uniquely adapted to retain total body K+ early in life. The magnitude and direction of net K+ transport in the cortical collecting duct (CCD), the segment responsible for the final renal regulation of K+ balance in the adult, reflect the balance of opposing fluxes of K+ secretion and K+ absorption. Evidence now indicates that the low capacity of the neonatal CCD for K+ secretion is due, at least in part, to a relative paucity of conducting K+ channels in the urinary membrane. A relative excess of K+ absorption in this nephron segment may further reduce net urinary K+ secretion. Under conditions prevailing in vivo, the balance of fluxes in the CCD likely contributes to the relative K+ retention characteristic of the neonatal kidney.

Absorption↗
PubMed2003

Chemical and enzymatic synthesis of fluorinated-dehydroalanine-containing peptides.

Michael acceptors have long been recognized as reactive functionalities that may link a biologically active molecule to its cellular target. 1,2-Dehydro amino acids are potential Michael acceptors present in a large number of natural products, but their reactivity is modulated by the deactivating nature of the alpha-amino group engaged in an amide bond. We describe here the preparation of 3-fluoro-1,2-dehydroalanine moieties within peptides that significantly enhance the reactivity of the Michael acceptor. Two different routes were designed to access these compounds, one relying on chemical means to introduce the desired functionality and the second taking advantage of a peptide epimerase. In the chemical approach, the fluoro-Pummerer reaction of cysteine derivatives afforded 3-fluorocysteine residues that were oxidized to the corresponding sulfoxides, followed by thermolytic elimination to provide the desired 3-fluorodehydroalanines. The mechanism of the fluoro-Pummerer reaction was investigated and several possible pathways were ruled out. The enzymatic approach utilized the dipeptide epimerase YcjG from Escherichia coli. Difluorinated alanine was incorporated at the C terminus of a dipeptide by chemical means. The resulting peptide proved to be a substrate for YcjG, which catalyzed fluoride elimination to provide the 3-fluorodehydroalanine-containing peptide. Mechanistic investigations showed that fluoride elimination occurred faster than epimerization and at a rate close to that of epimerization of Ala-Ala.

Alanine↗
PubMed2003

Biomimetic studies on the mechanism of stereoselective lanthionine formation.

Selenocysteine derivatives are useful precursors for the synthesis of peptide conjugates and selenopeptides. Several diastereomers of Fmoc-3-methyl-Se-phenylselenocysteine (FmocMeSec(Ph)) were prepared and used in solid phase peptide synthesis (SPPS). Once incorporated into peptides, the phenylselenide functionality provides a useful handle for the site and stereospecific introduction of E- or Z-dehydrobutyrine residues into peptide chains via oxidative elimination. The oxidation conditions are mild, can be performed on a solid support, and tolerate functionalities commonly found in peptides, including variously protected cysteine residues. Dehydropeptides containing unprotected cysteine residues undergo intramolecular stereoselective conjugate addition to afford cyclic lanthionines and methyllanthionines, which have the same stereochemistry as found in lantibiotics, a family of ribosomally synthesized and post-translationally modified peptide antibiotics. The observed stereoselectivity is shown to originate from a kinetic rather than a thermodynamic preference.

Alanine↗
PubMed2003

A copper-regulated transporter required for copper acquisition, pigmentation, and specific stages of development in Drosophila melanogaster.

The trace element copper is required for normal growth and development, serving as an essential catalytic co-factor for enzymes involved in energy generation, oxidative stress protection, neuropeptide maturation, and other fundamental processes. In yeast and mammals copper acquisition occurs through the action of the Ctr1 family of high affinity copper transporters. Here we describe studies using Drosophila melanogaster to investigate the role of copper acquisition through Ctr1 in normal growth and development. Three distinct Drosophila Ctr1 genes (Ctr1A, Ctr1B, and Ctr1C) have been identified, which have unique expression patterns over the course of development. Interestingly, Ctr1B, which is expressed exclusively during the late embryonic and larval stages of development, is transcriptionally activated in response to nutritionally induced copper deprivation and down-regulated in response to copper adequacy. The generation of Ctr1B mutant flies results in decreased larval copper accumulation, marked body pigmentation defects that parallel defects in tyrosinase activity, and specific developmental arrest under conditions of both nutritional copper limitation and excess. These studies establish that copper acquisition through the Drosophila Ctr1B transporter is crucial for normal growth and in early and specific stages of metazoan development.

Amino Acid Motifs↗
PubMed2003

Immunogene therapy of tumors with vaccine based on xenogeneic epidermal growth factor receptor.

The breaking of immune tolerance against self epidermal growth factor receptor (EGFr) should be a useful approach for the treatment of receptor-positive tumors with active immunization. To test this concept, we constructed a plasmid DNA encoding extracellular domain of xenogeneic (human) EGFr (hEe-p) or corresponding control mouse EGFr (mEe-p) and empty vector (c-p). Mice immunized with hEe-p showed both protective and therapeutic antitumor activity against EGFr-positive tumor. Sera isolated from the hEe-p-immunized mice exhibited positive staining for EGFr-positive tumor cells in flow cytometric analysis and recognized a single 170-kDa band in Western blot analysis. Ig subclasses responded to rEGFr proteins were elevated in IgG1, Ig2a, and Ig2b. There was the deposition of IgG on the tumor cells. Adoptive transfer of the purified Igs showed the antitumor activity. The increased killing activity of CTL against EGFr-positive tumor cells could be blocked by anti-CD8 or anti-MHC class I mAb. In vivo depletion of CD4(+) T lymphocytes could completely abrogate the antitumor activity, whereas the depletion of CD8(+) cells showed partial abrogation. The adoptive transfer of CD4-depleted (CD8(+)) or CD8-depleted (CD4(+)) T lymphocytes isolated from mice immunized with hEe-p vaccine showed the antitumor activity. In addition, the increase in level of both IFN-gamma and IL-4 was found. Taken together, these findings may provide a new vaccine strategy for the treatment of EGFr-positive tumors through the induction of the autoimmune response against EGFr in a cross-reaction between the xenogeneic homologous and self EGFr.

Adoptive Transfer↗
PubMed2003

[The utility boiler low NOx combustion optimization based on ANN and simulated annealing algorithm].

With the developing restrict environmental protection demand, more attention was paid on the low NOx combustion optimizing technology for its cheap and easy property. In this work, field experiments on the NOx emissions characteristics of a 600 MW coal-fired boiler were carried out, on the base of the artificial neural network (ANN) modeling, the simulated annealing (SA) algorithm was employed to optimize the boiler combustion to achieve a low NOx emissions concentration, and the combustion scheme was obtained. Two sets of SA parameters were adopted to find a better SA scheme, the result show that the parameters of T0 = 50 K, alpha = 0.6 can lead to a better optimizing process. This work can give the foundation of the boiler low NOx combustion on-line control technology.

Algorithms↗