Search

To determine the role of glycine (Gly) in the generation and modulation of basic respiratory rhythm.Neonatal (0-3 days) SD rats of either sex were used in this study. The medulla oblongata brain slice containing the medial region of the nucleus retrofacialis (mNRF) and the hypoglossal nerve rootlets was prepared, and the surgical procedure was performed in the modified Kreb's solution (MKS) with continuous carbogen (95% O(2) and 5% CO(2)) within 3 min. The rhythmical respiratory discharge activity (RRDA) of the hypoglossal nerve rootlets was recorded using suction electrode. Eighteen medulla oblongata slice preparations were divided into 3 groups and treated for 20 min with Gly receptor specific agonist Gly (10 micromol/L), Gly receptor antagonist strychnine (STR, 1 micromol/L), or Gly+STR after a 20 min Gly application. The changes in RRDA of the hypoglossal nerve rootlets were observed.Gly significantly decreased the inspiratory time and integral amplitude (IA), but the changes of respiratory cycle (RC) and expiratory time (TE) were not statistically significant. STR induced a decrease in expiratory time and respiratory cycle without significantly affecting the inspiratory time or integral amplitud. The effect of Gly on the respiratory rhythm was partially reversed by additional application of STR.Gly may play an important role in the modulation of RRDA in the medulla oblongata slice of neonatal rats.

To compare the differences in clinical therapeutic effect and safety between amphotericin B and its liposome form in treating invasive fungal infection (IFI) in hematological disorder with neutrocytopenia.Of 111 patients with IFI, 82 were treated with amphotericin B and 29 with amphotericin B liposome. The mean cumulative dose of amphotericin B was 617 (60-1895) mg and the mean course was 18 (7-60) d, and those for amphotericin B liposome was 925 (140-3420) mg and 13 (7-50) d, respectively.The total effective rates of amphotericin B and its liposome groups were 69% and 58%, respectively (P0.05). The adverse effect rates of chill and fever in amphotericin B and its liposome groups were 21% and 10% (P0.05), hypopotassemia 34% and 14% (P=0.03), hepatic impairment 22% and 17% (P0.05), and renal impairment 9% and 3%, respectively (P0.05).The therapeutic effect for IFI of amphotericin B and its liposome was similar. The severe adverse reaction of amphotericin B liposome was slightly lower than that of amphotericin B.

Altered glycosylation may be a hallmark of malignant transformation and cancer progression. In the work described, a specific mannose-binding lectin, Pinellia pedatisecta agglutinin (PPA), was genetically fused with the extracellular domain of coxsackie-adenovirus receptor (CAR) to generate the soluble CAR (sCAR)-PPA fusion protein. The adenoviral transduction of acute myeloid leukemia (AML) cell lines Kasumi-1 and HL-60 was increased by sCAR-PPA, indicating that a fraction of AML cells exposing mannose residues was detected by PPA. However, sCAR-PPA did not increase the adenoviral infection of KG-1 cells, suggesting the mannose exposure of AML cells may be cell type specific. Furthermore, the infectious efficiency of Ad-EGFP in chronic myeloid leukemia cell line K562 was significantly increased by sCAR-PPA as well. We, herein, report that PPA recognized a fraction of myeloid leukemia cells showing mannose-exposing phenotype. The sCAR-PPA fusion protein combined with the adenoviral vector system may provide a useful tool for investigating myeloid leukemia cells exposing mannose residues and further elucidating the role of these cells in the leukemia development.

The present study was carried out to determine the role of histamine H(1) and H(2) receptors in the generation of basic respiratory rhythm. Neonatal (aged 0-3 d) Sprague-Dawley rats of either sex were used. The medulla oblongata slice containing the medial region of the nucleus retrofacialis (mNRF) and the hypoglossal nerve rootlets was prepared and the surgical procedure was performed in the modified Kreb's solution (MKS) with continuous carbogen (95% O(2) and 5% CO(2)), and ended in 3 min. Respiratory rhythmical discharge activity (RRDA) of the rootlets of hypoglossal nerve was recorded by suction electrode. Thirty medulla oblongata slice preparations were divided into 5 groups. In groups I, II and III, histamine (5 μmol/L), H(1) receptor specific antagonist pyrilamine (10 μmol/L) and H(2) receptor specific antagonist cimetidine (5 μmol/L) was added into the perfusion solution for 15 min separately. In group IV, after application of histamine for 15 min, additional pyrilamine was added into the perfusion for another 15 min. In group V, after application of histamine for 15 min, additional cimetidine was added into the perfusion for another 15 min. The discharges of the roots of hypoglossal nerve were recorded. Signals were amplified and band-pass filtered (100-3.3 kHz). Data were sampled (1-10 kHz) and stored in the computer via BL-420 biological signal processing system. Our results showed that histamine significantly decreased the respiratory cycle (RC) and expiratory time (TE), but changes of integral amplitude (IA) and inspiratory time (TI) were not statistically significant. Pyrilamine induced significant increases in RC and TE, but changes of TI and IA were not statistically significant. Cimetidine had no effects on RC, TE, TI and IA of RRDA. The effect of histamine on the respiratory rhythm was reversed by additional application of pyrilamine but not cimetidine. Taken together, with the results mentioned above, histamine H(1) receptors but not H(2) receptors may play an important role in the modulation of RRDA in the medulla oblongata slice preparation of neonatal rats.

To observe the effects of Tangweian granule on 5-HT(2A)R in rat model with diabetic gastroparesis (DGP).The rats with diabetic gastroparesis induced by injecting alloxan and giving 200% Radix Rehmanniae preparata were divided into four groups randomly: Tangweian high dosage group, Tangweian low dosage group, motilium control group and the model control group, 10 rats each group. Each group was irrigated with drugs during establishing the model. Additionally, we chose 10 rats by way of normal control group. Further more, Tangweian high dosage group were irrigated stomach with gliclazide 20 mg x kg(-1) and Tangweian granule 31.75 g x kg(-1); Tangweian low dosage group were irrigated stomach with gliclazide 20 mg x kg(-1) and Tangweian granule 15.88 g x kg(-1); motilium control group were irrigated stomach with gliclazide 20 mg x kg(-1) and motilium 3.75 mg x kg(-1) and the model control group were irrigated stomach with distilled water. Then the effects of Tangweian granule on 5-HT(2A)R were observed.The curative group had better effects than the control group in lowering the blood sugar and the level of 5-HT(2A)R content (P0.01). And there was significant difference between the curative group and control group (P0.05).It is verified that Tangweian granule has obvious effects on lowering the blood sugar and improving the level of 5-HT(2A)R.

To analyse the outcome of newly diagnosed adult acute myeloid leukemia (AML) patients treated with HAA (homoharringtonine, cytarabine and aclarubicin) regimen and explore the efficacy and safety of this regimen.Eighty patients were treated with HAA regimen. The complete remission (CR) rate was observed. Kaplan-Meier method was used to estimate relapse free survival (RFS) rate and the differences were compared with 2-sided log-rank test.Of the 80 patients, 65 (81%) attained CR and the CR rate after the first course of induction was 75%. For the CR patients, the median follow-up was 26 (2 -69) months, and the estimated 3-year overall survival (OS) rate was 51% and the estimated 3-year RFS was 53%. For the AML-M5 and AML-M /M2 patients the CR rate was 74% and 87% and 3 year RFS of CR patients was 75% and 37%, respectively. The CR rate of 100%, 83% and 20% was achieved in patients with favorable, intermediate and unfavorable cytogenetics, respectively. The 3 year OS for favorable and intermediate group was 76% and 50% respectively. The median survival time of unfavorable group was only 6 months.HAA regimen is a safe, efficacious, and well-tolerable induction therapy for newly diagnosed AML.

The flow shop scheduling problem (FSSP) with limited buffers constraint is a typical NP-hard combinatorial optimization problem and represents an important area in production scheduling. In this paper, a class of differential evolution (DE) method with the optimal computing budget allocation (OCBA) technique and hypothesis test (HT), namely OHTDE, is proposed for the stochastic flow shop scheduling with limited buffers between consecutive machines to minimize the maximum completion time (i.e., makespan). In the OHTDE, the population-based search mechanism of DE and a special crossover are applied for well exploration and exploitation, and the OCBA technique is used to allocate limited sampling budgets to provide reliable evaluation and identification for good individuals. Meanwhile, HT is also applied to perform a statistical comparison to avoid some repeated search to some extent. The results and comparisons demonstrate the superiority of OHTDE in terms of effectiveness and robustness.

To investigate the effects of nikethamide on the generation and modulation of rhythmic respiration of neonatal rats and the role of 5-HT(2A) receptor in this course, experiments were performed on the transverse medullary slices of neonatal rats (both sexes, 1-3 d) in vitro. The slices containing the medial region of the nucleus retrofacialis (mNRF) with the hypoglossal nerve rootlets were prepared in which the respiratory-related rhythmic discharge activity (RRDA) was recorded from the hypoglossal nerve rootlets by suction electrode. The possible role of nikethamide on RRDA was investigated by administration of an agonist of 5-HT(2A) receptor, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), and an antagonist of 5-HT(2A) receptor, ketanserine, dissolved in modified Krebos solution (MKS). Thirty slices were randomly divided into five groups: Group 1: the slices were perfused with different concentrations of nikethamide (0.5, 1, 3, 5, 7, 10 μg/mL), and the most effective concentration was selected; Group 2: the slices were perfused with DOI (40 μmol/L); Group 3: the slices were perfused with ketanserine (40 μmol/L); Group 4: the slices were perfused with ketanserine + DOI; Group 5: the slices were perfused with nikethamide, then perfused with nikethamide + ketanserine after washout of nikethamide. Nikethamide increased RRDA in transverse medullary slices at 0.5-7 μg/mL, and 5 μg/mL was the most effective concentration. DOI increased RRDA with prolonged inspiratory time (TI), increased integral amplitude (IA), and shortened respiratory cycle (RC). Ketanserine decreased RRDA with shortened TI, decreased IA and prolonged RC. Ketanserine + DOI had no significant effects on RRDA. The effects of nikethamide on RC and IA were totally and partially reversed by additional application of ketanserine, but the effect of nikethamide on TI was not influenced by ketanserine. It is proposed that nikethamide increases RRDA partly via 5-HT(2A) receptors.

To detect the expression levels of telomere binding factor 2 (TRF2) on leukemia cell lines and primary leukemia cells.The expression of TRF2 mRNA was detected with quantitative real-time RT-PCR in leukemia cell lines and primary leukemia cells. The Western blot analysis was used for the detection of TRF2 protein expression.TRF2 was overexpressed in T-cell leukemia cell lines but not in myelogenous leukemia cell lines. Significant higher expression levels of TRF2 were observed in primary leukemia cells from patients with M0 and M1 subtypes of acute myelogenous leukemia (AML) compared with normal control and other subtypes of AML.Increased TRF2 expression levels are found in T-cell leukemia cell lines and AML patients with poor prognosis, which suggests that TRF2 expression might be related to the prognosis of leukemia.

To investigate the role of GABA A receptor in nikethamide-induced respiratory enhancement in the medullary slices of neonatal rats.Ex vivo medullary slices of neonatal rats (1 to 3 days old) containing the medial region of the nucleus retrofacialis with the hypoglossal nerve rootlets were prepared and perfused with modified Kreb's solution to record respiration-related rhythmic discharge activity (RRDA) from the hypoglossal nerve rootlets using suction electrodes. Thirty RRDA-positive slices were randomized into 5 equal groups and perfused with nikethamide (at concentrations of 0.5, 1, 3, 5, 7, and 10 microg/ml with the optimal nikethamide concentration determined), GABA (at 10, 20, 40, and 60 micromol/ to determine the optimal concentration), 10 micromol/ bicuculline, 10 micromol/ bicuculline plus 40 micromol/L GABA, and 5 microg/ml nikethamide followed by 5 microg/ml nikethamide plus 10 micromol/ bicuculline after wash out, respectively.Nikethamide increased RRDA at the concentrations of 0.5-7 microg/ml, and 5 microg/ml nikethamide showed the most distinct effect on the inspiratory time (TI), integral amplitude (IA), and respiratory cycle (RC). GABA at 40 micromol/ showed the most effective inhibition of RRDA in terms of TI, IA, and RC. Bicuculline at 10 micromol/ could increase the IA, TI and RC, but the combination of 10 micromol/ bicuculline and 40 micromol/ GABA had no significant effects on RRDA. Compared with nikethamide used alone, nikethamide plus bicuculline significantly increased TI and IA without affecting RC.Nikethamide can enhance RRDA of the hypoglossal nerve rootlets in the medullary slices of neonatal rats, and the effect can be partially mediated by the GABA A receptor.

Aims This study was designed to investigate the effects of sodium ferulate (SF) on rat isolated thoracic aortas and the possible mechanisms. Main methods Isometric tension was recorded in response to drugs in organ bath. Cytosolic free Ca 2+ concentration ([Ca 2+ ] i ) was measured using Fluo-3 in cultured rat aortic smooth muscle cells (RASMC). Key findings SF (0.1–30 mM) relaxed the isolated aortic rings precontracted with phenylephrine (PE) and high-K + in a concentration-dependent manner with respective pD 2 of 2.7 ± 0.02 and 2.6 ± 0.06. Mechanical removal of endothelium did not significantly modify the SF-induced relaxation. In Ca 2+ -free solution, SF noticeably inhibited extracellular Ca 2+ -induced contraction in high-K + and PE pre-challenged rings, and suppressed the transient contraction induced by PE and caffeine. The vasorelaxant effect of SF was unaffected by various K + channel blockers such as tetraethylammonium, glibenclamide, 4-aminopyridine, and barium chloride. In addition, SF concentration-dependently reduced the contraction induced by phorbol-12-myristate-13-acetate, an activator of protein kinase C (PKC), in the absence of extracellular Ca 2+ , with the pD 2 of 2.9 ± 0.03. In RASMC, SF had no effect on PE- or KCl-induced [Ca 2+ ] i increase either in the presence or in the absence of external Ca 2+ . Significance These results indicate that SF acts directly as a non-selective relaxant to vascular smooth muscle. The direct inhibition of the common pathway after [Ca 2+ ] i increase may account for the SF-induced relaxation in Ca 2+ -dependent contraction, while the blockage of the PKC-mediated contractile mechanism is likely responsible for the SF-induced relaxation in Ca 2+ -independent contraction.

One of the key challenges in computational biology is prediction of three-dimensional protein structures from amino-acid sequences. For most proteins, the "native state" lies at the bottom of a free-energy landscape. Protein structure prediction involves varying the degrees of freedom of the protein in a constrained manner until it approaches its native state. In the Rosetta protein structure prediction protocols, a large number of independent folding trajectories are simulated, and several lowest-energy results are likely to be close to the native state. The availability of hundred-teraflop, and shortly, petaflop, computing resources is revolutionizing the approaches available for protein structure prediction. Here, we discuss issues involved in utilizing such machines efficiently with the Rosetta code, including an overview of recent results of the Critical Assessment of Techniques for Protein Structure Prediction 7 (CASP7) in which the computationally demanding structure-refinement process was run on 16 racks of the IBM Blue Gene/L (TM) system at the IBM T. J. Watson Research Center. We highlight recent advances in high-performance computing and discuss,future development paths that make use of the next-generation petascale (> 10(12) floating-point operations per second) machines.

A blind image watermarking scheme based on secret sharing in discrete wavelet transform domain is proposed. Watermark was divided into n shadows according to secret sharing scheme. And t or more of those shadows can reconstruct the watermark, while t-1 or less shadows could not do it. In order to achieve optimum embedding strategy, a closed loop embedding process is proposed, which is modified iteratively according to results of performance analysis. The convergence of closed loop watermarking is proved. Independent component analysis is utilized so that detector can not merely detect watermark but also can extract it. Before watermark reconstruction, one way hashing function is used to withstand cheating attacks. The experimental results show that it is robust against a wide range of attacks proposed by Stirmark and it is more safety than traditional watermarking techniques.

The human proteome is rich with protein kinases, and this richness has made the kinase of crucial importance in initiating and maintaining cell behavior. Elucidating cell signaling networks and manipulating their components to understand and alter behavior require well designed inhibitors. These inhibitors are needed in culture to cause and study network perturbations, and the same compounds can be used as drugs to treat disease. Understanding the structural biology of protein kinases in detail, including their commonalities, differences and modes of substrate interaction, is necessary for designing high quality inhibitors that will be of true use for cell biology and disease therapy. To this end, we here report on a structural analysis of all available active-conformation protein kinases, discussing residue conservation, the novel features of such conservation, unique properties of atypical kinases and variability in the context of substrate binding. We also demonstrate how this information can be used for structure prediction. Our findings will be of use not only in understanding protein kinase function and evolution, but they highlight the flaws inherent in kinase drug design as commonly practiced and dictate an appropriate strategy for the sophisticated design of specific inhibitors for use in the laboratory and disease therapy.

We describe predictions made using the Rosetta structure prediction methodology for both template-based modeling and free modeling categories in the Seventh Critical Assessment of Techniques for Protein Structure Prediction. For the first time, aggressive sampling and all-atom refinement could be carried out for the majority of targets, an advance enabled by the Rosetta@home distributed computing network. Template-based modeling predictions using an iterative refinement algorithm improved over the best existing templates for the majority of proteins with less than 200 residues. Free modeling methods gave near-atomic accuracy predictions for several targets under 100 residues from all secondary structure classes. These results indicate that refinement with an all-atom energy function, although computationally expensive, is a powerful method for obtaining accurate structure predictions.

A challenge in protein–protein docking is to account for the conformational changes in the monomers that occur upon binding. The RosettaDock method, which incorporates sidechain flexibility but keeps the backbone fixed, was found in previous CAPRI rounds (4 and 5) to generate docking models with atomic accuracy, provided that conformational changes were mainly restricted to protein sidechains. In the recent rounds of CAPRI (6–12), large backbone conformational changes occur upon binding for several target complexes. To address these challenges, we explicitly introduced backbone flexibility in our modeling procedures by combining rigid-body docking with protein structure prediction techniques such as modeling variable loops and building homology models. Encouragingly, using this approach we were able to correctly predict a significant backbone conformational change of an interface loop for Target 20 (12 A rmsd between those in the unbound monomer and complex structures), but accounting for backbone flexibility in protein–protein docking is still very challenging because of the significantly larger conformational space, which must be surveyed. Motivated by these CAPRI challenges, we have made progress in reformulating RosettaDock using a “fold-tree” representation, which provides a general framework for treating a wide variety of flexible-backbone docking problems. Proteins 2007. © 2007 Wiley-Liss, Inc.

Overexpression of human telomere repeat binding factor 2 (TRF2), which may play an important role in the fate of cancer cells, has been observed in adult T-cell leukemia. Previous reports have shown that the inhibition of TRF2 results in the apoptosis of cancer cells. In this study, we demonstrated that arsenic trioxide (As2O3) induced in vitro growth inhibition and/or apoptosis of human T-cell leukemia cell line Molt-4 in a caspase-independent manner. Telomerase activity was not inhibited, although the level of the reverse transcriptase subunit of the human telomerase gene (hTERT) mRNA expression was down regulated during the early times and then recovered to the level found in untreated controls about 48 hours after treatment with As2O3. Furthermore, a remarkable telomere shortening related to exposure of As2O3 was observed in 50 population doubling. Inc ontrast, the alteration of telomere length did not occur after exposure to higher concentration of As2O3 (10 microM) for 24 hours and 48 hours, respectively, suggesting that the shortening of telomeres induced by As2O3 is dependent of a series of cell division cycles. Chromosomal analysis showed that As2O3 exposure caused chromosomal end-to-end fusion in human T-cell leukemia cells while downregulation of TRF2 was observed. Finally, the inhibition of TRF2 protein expression and the sensitivity to As2O3 in a panel of leukemia cell lines were checked. The data revealed that inhibition of TRF2 rendered leukemia cells more susceptible to As2O3. In conclusion, the downregulation of TRF2 by As2O3 contribute to chromosomal end-to-end fusion, and apoptosis in leukemia cells, suggesting that TRF2 could be an attractive target for new therapies of leukemia.

For the purpose of estimating DOAs and delays of multipath signals, a novel improved 2-dimensional (2-D) ESPRIT is proposed. The improved method estimates the joint space-time signal subspace firstly. Then FFT and deconvolution is done to map time delay parameters into frequency phase shifts and a modified joint signal subspace is constructed. At last DOAs and delays are estimated by using rotational invariance property. Compared with traditional 2-D ESPRIT method, the method we proposed here can prevent the correlated noise problem when FFT is done to estimated channel directly. Simulation results show the effectiveness of the improved method.

To investigate the effects of ST1571 on the development of dendritic cells (DC) derived from bone marrow mononuclear cells of patients with chronic myeloid leukemia (CML).Bone marrow mononuclear cells (BMMNC) from CML patients and healthy volunteers were cultured initially using multiple cytokine combinations as follows: recombinant human granulocyte/ macrophage colony-stimulating-factor (rhGM-CSF) plus recombinant human interleukin-4 (rhIL-4) as CML and normal control groups, rhGM-CSF plus rhIL-4 and ST1571 as CML experimental groups, and from day 8 recombinant human tumor necrosis factor-alpha ( rhTNF-alpha) was added to stimulate DC maturation. The morphologic features of cells were observed by Wright's staining and phenotypes were assessed by flow cytometry. Cytogenetic analysis was performed by fluorescence in-situ hybridization (FISH), and the antigen-presenting function was assayed by mixed lymphocyte reaction (MLR). The concentration of VEGF was detected by ELISA.CML experimental groups treated with STI571 displayed morphological features similar to those of control groups with delicate membrane projections. However, in comparison with the CML control groups, the CML experimental groups showed an increased expression of CD80, CD86, CD83 and HLA-DR and showed more intense abilities of allogeneic antigen presentation, which were similar to those of normal control groups. FISH confirmed that DCs of both CML, groups were of leukemic origin. The concentration of VEGF was dramatically reduced in CML experimental groups.In vitro, STI571 promotes the activation/maturation of DCs derived from BMMNCs of patients with CMI, and decreases VEGF production by the leukemic cells. The promotion of DC maturation may be partially due to decreased inhibitory effect of VEGF.

The melanoma differentiation-associated gene-7/interleukin-24 (MDA-7/IL-24) is a novel candidate of tumor suppressor, which can experimentally inhibit the proliferation of cancer cells and induce apoptosis of various human malignant cells.

For large-scale chemical process, which consists of lots of production units, all units have their respective optimization objects which are often conflicting with each other for a series of constraints on material and energy balance. In this paper, the total solution with two layers structure strategy made up of multi-units unified optimization and predictive control of each unit is realized. For the global optimization has high dimension, serious nonlinearity and uncertainty, the optimization algorithm based on differential evolution (DE) is performed, while a hybrid DE approach combining hypothesis test (HT) to compare the optimization objects under uncertainty is proposed. The simulation results of an application example to a 20Mt/a gas separation process show that the proposed total solution with two layers structure strategy is successful and multi-units unified optimization method based on HTDE is effective and robust for solving the optimization problem under uncertainty.

To study the expression of epithelial-cadherin (E-cad), CD44v6 and Cx43 in hepatocellular carcinoma (HCC) and its relationship with sex and age of patients, as well as tumor histopathologic grades.Double immunofluorescent staining and laser scanning confocal microscopy was used to study the expression of E-cad, CD44v6 and Cx43 in 30 cases of normal liver tissue, 25 cases of benign hepatic lesions and 38 cases of HCC. In the HCC group, correlation of antigen expression with sex and age of patients and tumor histopathologic grades was studied by T-test.Significant decrease in expression of E-cad and Cx43 was noted in HCC group, as compared to normal liver tissue and benign hepatic lesion (P0.05). On the other hand, CD44v6 expression was higher in HCC group than in the other two groups (P0.05). In HCC group, the expression of E-cad and Cx43 did not correlate with sex, age and histopathologic grades (P0.05). However, CD44v6 expression positively correlated with higher tumor histopathologic grades (P0.05) but not sex and age of patients (P0.05). In HCC group, the expression of E-cad positively correlated with that of Cx43, while the expression of CD44v6 negatively correlated with that of E-cad and Cx43.Tumor immunophenotype alters during development and progression of HCC. Low expression of E-cad and Cx43 and high expression of CD44v6 may be related to aggressive clinical behavior of HCC, moreover, high expression of CD44v6 correlated with high tumor grades. Detection of E-cad, CD44v6 and Cx43 expression may thus be useful in predicting prognosis of HCC.

This paper proposes an effective Differential Evolution (DE) based hybrid algorithm for Multi-objective Permutation Flow Shop Scheduling Problem (MPFSSP), which is a typical NP-hard combinatorial optimization problem. In the proposed Multi-objective Hybrid DE (MOHDE), both DE-based searching operators and some special local searching operators are designed to balance the exploration and exploitation abilities. Firstly, to make DE suitable for solving MPFSSP, a largest-order-value (LOV) rule based on random key representation is developed to convert the continuous values of individuals in DE to job permutations. Then, to enrich the searching behaviors and to avoid premature convergence, a Variable Neighborhood Search (VNS) based local search with multiple different neighborhoods is designed and incorporated into the MOHDE. Simulation results and comparisons with the famous random-weight genetic algorithm (RWGA) demonstrate the effectiveness and robustness of our proposed MOHDE.

To construct expression vector of hTERT-hIL-18 fusion gene in eukaryotic cells and to study its biological function.hIL-18 gene was amplified by RT-PCR, then T-A cloned and inserted into PCDNA3.1(+)/hTERT vector. The sequence of fusion gene was examined by enzyme incision and DNA sequencing. The vector with fusion gene was transformed into 3T3 cells by the method of lipofecting, and proved by Western blot. The secretion gamma-interferon was measured with ELISA and cell apoptosis was detected with flow cytometry.Expression vector PCDNA3.1(+) of hTERT/hIL-18 fusion gene was constructed successfully. The correct sequence was proved by enzyme incision and sequencing and there was a correct open reading frame. Fusion protein of hTERT/hIL-18 was effectively expressed in eukaryotic cells and was proved by Western blot and immunofluorescence stain. The fusion protein stimulated KG-1 cells to secrete gamma-interferon and had anti-apoptosis effect.Fusion protein hTERT-hIL-18 is highly effectively expressed in eukaryotic cells and is biologically active.

To investigate the effect of topotecan (TPT) on human myelodysplastic syndrome (MDS) cells in vitro and in vivo.Cell growth was measured by a MTT assay. The percentage of cells undergoing apoptosis was determined by flow cytometry after staining with annexin V-FITC and propidium iodide. The morphology of apoptotic cells was observed by transmission electron microscopy (TEM). Furthermore, the antitumor effect on MDS cells in xenotransplanted severe combined immunodeficiency (SCID) mice was evaluated by tumor volume and survival. Western blot was used for determining the expression of topoisomerase I (Top1) protein.The growth of Mutz-1 cells was suppressed by TPT treatment in a dose-dependent manner. The 50% inhibition in Mutz-1 cell growth (IC(50)) of TPT for 72 h was 272 ng/L. The percentage of apoptotic cells observed in the Mutz-1 cells after exposure to TPT (160 ng/L) in 48 h and 72 h was (54.16 +/-4.29)% and (72.97+/-6.12)%, respectively. TEM showed the characteristics of apoptosis in Mutz-1 cells treated with TPT. The xenotransplanted SCID mice treated with TPT showed inhibited tumor growth compared with control group. TPT treatment resulted in a longer survival as compared with the control group (P0.001) and with the As2O3-treated group (P0.001). The cells exposed to TPT exhibited a time-dependent decrease of Top1 protein expression.TPT can inhibit Mutz-1 cell growth and induce apoptosis in vitro.The downregulation of Top1 may be involved in the apoptosis induced by TPT. TPT has a significant antitumor effect in vivo.

In this paper, adaptive fuzzy control is proposed for a class of uncertain MIMO nonlinear systems with unknown dead-zones and a triangular control structure. The design is based on the principle of sliding mode control and the property of Nussbaum function. The approach does not require a priori knowledge of the sign of the control gain and the upper bound and lower bound of dead zone model parameter to be known a priori. The adaptive compensation for the optimal approximation error is employed to minimize the influence of modeling errors and parameter estimation errors. By theoretical analysis, the closed-loop control system is proved to be stable in the sense that all signals involved are bounded, with tracking errors converging to zero. Simulation results demonstrate the effectiveness of the approach.

To investigate the telomerase activity and the telomere length in CD4(+), CD8(+) and CD19(+) lymphocytes from patients with systemic lupus erythematosus (SLE).Telomerase activity of CD4(+), CD8(+) and CD19(+) cells from patients with SLE and normal controls was examined by telomeric repeats amplification protocol. Telomere length in those cells was measured by Southern blot method.CD4(+), CD8(+) and CD19(+) cells in patients with SLE showed high telomerase activity, but only telomerase activity of CD19(+) cells was significantly higher than that in controls. The length of telomere in CD4(+) and CD8(+) cells was significantly shorter than that in controls, but not in CD19(+) cells.Higher telomerase activity in CD19(+) cells and shortened telomere length in CD4(+) and CD8(+) cells of patients with SLE may be associated with pathogenesis of SLE.

To investigate the mechanisms of the telomerase regulations during the apoptosis of the human MDS-RAEB cell line MUTZ-1 cells induced by arsenic trioxide (As(2)O(3)), telomerase activity was detected by TRAP-ELISA and the expressions of mRNAs of hTERT, TRF1 (TTAGGG repeat binding factor 1), TRF2 (TTAGGG repeat binding factor 2), bcl-2, and bax genes were detected by RT-PCR. Apoptosis was detected by translocation of phosphatidylserine (PS) by flow cytometry. The results showed that 1 - 8 micromol/L of As(2)O(3) induced typical apoptosis of MUIZ-1 cells in the dose-and time-dependent manners, the telomerase activity could be down-regulated at this concentration and negatively correlated with increased apoptosis (r = -0.938, P = 0.018). The expression of telomerase activity was positively related to the expression of hTERT (r = 0.783, P = 0.022), but As(2)O(3) had no effect on the mRNA expression of TRF1 and TRF2 genes. The inhibition of telomerase activity by As(2)O(3) on MUTZ-1 cells was accompanied with the low expression of bcl-2 gene and the decrease of bcl-2/bax ratio. It is concluded that the apoptosis of MUTZ-1cells induced by As(2)O(3) may occur via the inhibition of telomerase activity and down-regulation of the expression of hTERT mRNA, and this may be one of the mechanisms inducing apoptosis in MUTZ-1 cells treated by As(2)O(3).

To induce primary chronic myeloid leukemia (CML) cells into dendritic cells (DCs).Bone marrow mononuclear cells (MNCs) were isolated from 13 CML patients and peripheral blood MNCs from 5 healthy donors. The isolated MNCs were co-cultured with rhGM-CSF 1,000 U/ml, rhIL- 4,500 U/ml and TNF-alpha 50 U/ml for 10 days. The morphological features were observed by Wright's staining,inverted microscope and electron microscope. CD(80), CD(86), CD(83), CD(1a) and HLA-DR expression were assayed by flow cytometry, cytogenetic analysis was performed by fluorescence in-situ hybridization(FISH). The concentration of IL-12 was measured by ELISA and the function of antigen presenting was tested by mixed lymphocyte reaction (MLR).After being cultured with cytokines, the typical dendritic appearance with delicate membrane projections was observed. The CD(80), CD(86), CD(83), CD(1a) and HLA-DR markers and capacity of stimulating allogeneic T cells were upregulated significantly. FISH confirmed that the DCs were generated from leukemic origin and CML DCs could secrete higher level of IL-12 than CML MNCs. There were no differences in morphology and immunophenotype expression between DCs derived from CML and those from normal individuals. However, DCs from CML patients displayed weaker activity than that of normal individuals when tested in MLR.CML cells could be induced into leukemia-DCs by co-culture with cytokines.

To investigate the effects of interleukin 7 (IL-7) on B7 molecules expression and immunogenicity of acute leukemia (AL) cells.The B7 molecules expression on fresh acute leukemia cells and on the IL-7 exposed leukemia cells was detected by FACScan cytometer. B7-1 and B7-2 mRNA in IL-7 treated HL-60 cells were detected by reverse transcription-PCR (RT-PCR). The stimulation of proliferation of allogeneic peripheral blood mononuclear cells (PBMNC) by IL-7 treated leukemia cells was detected by MTT method. The level of interferon-gamma (IFN-gamma) secreted by the stimulated PBMNC was determined using enzyme-linked immunosorbent assays (ELISA). The blocking experiments were performed using monoclonal antibodies against B7-1, B7-2 and W6/32.B7-1 was weakly expressed in three, whereas B7-2 did in only one of eleven AL patients. IL-7 significantly enhanced B7 molecules expression on AL cells in a time-dependent manner. Furthermore, IL-7 could induce higher expression of B7-1 and B7-2 mRNAs on HL-60 cells. IL-7 treated leukemia cells could stimulate PBMNC proliferation and promote their IFN-gamma production. Anti-B7-1 and anti W6/32 but not anti-B7-2 monoclonal antibodies significantly inhibited the stimulated PBMNC proliferation and IFN-gamma secretion.Fresh AL cells express low level of B7-1 and B7-2 molecules. IL-7 enhances the B7 molecules expression on AL cells. The IL-7-treated leukemia cells can significantly stimulate the proliferation of allogeneic PBMNC and induce their IFN gamma secretion.

A new concept of ‘W’ tailless aerodynamic configuration was presented in this paper. This configuration is designed based on the flow behavior of forward-swept wing (FSW) and the flow control concept. It adopts blended high swept side-strake body design technique and adapts to high subsonic and maneuverability aircraft. By numerical simulation, Flow mechanism and aerodynamic characteristic of ‘W’ configuration and referenced FSW configuration are investigated. ‘W’ configuration shows excellent longitudinal aerodynamic performance. Both of yaw and roll are unstable, however, the moment values are small. The roll stability can be achieved above medium angle of attack, and yaw stability can be improved with proper aerodynamic ways by use of FSW’s favorable outboard wing stall performance. The results show that ‘W’ tailless configuration takes on excellent aerodynamic characteristics. Its design idea is reasonable and has a good develop foreground.

The classification of the uterine myoma and adenomyosis from their ultrasound images mainly depends on doctors' experience and lacks objective criterions. Here a novel classification method is proposed using the multiresolution analysis and the orientational fractal analysis. Firstly, texture features under various resolutions and orientational fractal features are obtained from ultrasound images. Then the feature selection (FS) process is implemented using the sequential forward selection algorithm (SFS). Finally a classifier based on the support vector machine (SVM) is set up to classify the images into normal (Nor), myoma (Myo) and adenomyosis (Ad) cases. From the application of 27 Nor, 45 Ad and 74 Myo cases, it is showed that orientational fractal features and some multiresolution texture features are sensitive to the uterine Myo and Ad classification. The SVM classifier with the selected features may be useful in the practical classification.

Accurate high-resolution refinement of protein structure models is a formidable challenge because of the delicate balance of forces in the native state, the difficulty in sampling the very large number of alternative tightly packed conformations, and the inaccuracies in current force fields. Indeed, energy-based refinement of comparative models generally leads to degradation rather than improvement in model quality, and, hence, most current comparative modeling procedures omit physically based refinement. However, despite their inaccuracies, current force fields do contain information that is orthogonal to the evolutionary information on which comparative models are based, and, hence, refinement might be able to improve comparative models if the space that is sampled is restricted sufficiently so that false attractors are avoided. Here, we use the principal components of the variation of backbone structures within a homologous family to define a small number of evolutionarily favored sampling directions and show that model quality can be improved by energy-based optimization along these directions. With the progression of structural genomics initiatives (1–3), comparative modeling has become an increasingly important method for building protein structure models (4, 5). After a suitable structure template is chosen, accurate comparative modeling requires a correct alignment between the target protein sequence and the template sequence, an accurate method for modeling the loops (the insertions and deletions in an alignment) and side chains, and, finally, a method for refining the coordinates derived from the template structure toward those of the true native structure (6–8). In this study, we focus on this last model-refinement step. Improvement of the accuracy of comparative models is very important because accurate comparative models potentially can be used for many applications, such as virtual drug scanning (9), molecular replacement (10), and function prediction (11). Refinement is particularly important when the sequence identity between a target protein and the template protein is 1.5 Å (13)., However, high-resolution refinement is as formidable as it is important. This difficulty is due to both the large size of conformational space and the delicate balance of forces in the native state. Indeed, in the recent CASP5 experiment (The 5th Community Wide Experiment on the Critical Assessment of Techniques for Protein Structure Prediction), most refined structures had larger rmsd to the native structure than the starting template backbone conformation (7). High-resolution refinement is thus a very stringent test of accuracy that perhaps no current force field satisfies. Progress on this very important but very challenging problem may be facilitated by focusing on more constrained and thus more tractable refinement problems. We were led to thinking about such problems by the observation that a refinement protocol that did not markedly improve de novo structure-prediction models was very much more successful on the more constrained rigid-body protein–protein docking problem (14). The greatly reduced number of backbone degrees of freedom in the protein–protein docking problem significantly reduces the number of false attractors in the free-energy landscape: As illustrated in ref. 14, the docking free-energy landscapes typically are funneled strongly into the native minimum. The conceptual step forward in this paper is to use evolutionary information to reduce the number of degrees of freedom in the monomeric protein-refinement problem to mimic the situation in the protein–protein docking problem. We accomplish this goal by restricting sampling to the subspace defined by the largest principal components (PCs) of the variation in the structural core of homologous proteins. This strategy greatly enhances the sampling of near-native backbone conformations, and the low-energy models identified by using the Rosetta high-resolution energy function (15, 16) usually have lower rmsd to the native backbones than the starting templates. This restricted refinement problem can provide a testing ground for evaluating and improving potential functions for the unrestricted comparative-modeling refinement problem. More practically, the refinement of structure cores by energy-based sampling along evolutionarily preferred directions can serve as the first step toward improving a model structure built from a template. After a more accurate structure core is obtained, the rest of the structure can be built by using loop modeling and side-chain repacking (6)., This work is supported by the Howard Hughes Medical Institute and Ministerio de Ciencia y Tecnología Grant BIO2001-3745 (to A.R.O.). Research at Centro Biología Molecular Severo Ochoa is facilitated by an institutional grant from the Ramón Areces Foundation

To detect the expression levels of telomere binding factor 2 (TRF2) mRNA in tumor tissue of non-Hodgkin lymphoma (NHL) patients using quantitative real-time RT-PCR.The target gene mRNA was amplified with RT-PCR, then was sequentially electrophoresed and purified as standards, and the standard curves of gene expression were established. The expression levels of TRF2 mRNA of lymphoid tissue from NHL and reactive lymphoadenopathy were detected with real-time RT-PCR.The correlation coefficient was 0.996 between the amount of template cDNA and the intensity of fluorescence signal when gene expression standard curves were established. The correlation coefficient of template cDNA amount and grey density of bands derived from gel electrophoresis of real-time RT-PCR final products was 0.779 (P0.05). Of all NHL patients, expression levels of TRF2 mRNA of follicular lymphoma, mantle cell lymphoma and diffuse large B cell lymphoma were(22.943 +/-9.424) amol, (23.181 +/-5.983) amol and (18.339 +/-7.910) amol, respectively, which had no significant difference compared with reactive lymphoadenopathy [(21.796 +/-4.800) amol, P0.05]. The expression level of TRF2 mRNA of Burkitt lymphoma was (33.170 +/-12.841) amol, which was significantly higher than that of reactive lymphoadenopathy and other types of NHL (P0.05).Alcohol drinking isn't one of the risk factors of colorectal cancer among Jiashan County population.

Motivation: Much information about new protein sequences is derived from identifying homologous proteins. Such tasks are difficult when the evolutionary relationships are distant. Some modern methods achieve better results by building a model of a set of related sequences, and then identifying new proteins that fit the model. A further advance was the development of iterative methods that refine the model as more homologs are discovered. These methods are generally limited by ad hoc methods of sequence weighting, neglect of underlying evolutionary relationships and the representation of the set with a single one-size-fits-all model. These limitations are avoided through the use of a Tree hidden Markov model (T-HMM) approach. Our previous work described how a non-iterative version of the T-HMM method could identify distant homologs with superior performance compared with other non-iterated approaches, and described how this method was particularly appropriate for being implemented as an iterative algorithm. Results: We describe an iterative version of the T-HMM algorithm, and evaluate its performance for the detection of distant homologs. Significant improvement over other commonly used methods is found. Availability: The software (C++, Perl) is available from the corresponding author.

Microsomal epoxide hydrolase (mEH) is a bifunctional protein that plays a crucial role in the metabolism of numerous xenobiotics as well as in mediating the hepatic sodium-dependent uptake of bile acids that are involved in numerous physiological processes including the regulation of cholesterol metabolism. The transcription factors and nuclear receptors that control the constitutive and inducible expression of the mEH gene (EPHX1), however, have not been described. To characterize these factors, a series of 5'-deletion constructs have been transfected into human liver-derived HepG2 cells as well as non-hepatic HeLa cells. Promoter activity analysis indicated the presence of a positive regulatory element in the -80/-70 bp region. Sequence analysis revealed a putative GATA site at -79/-74 bp as well as an additional site at -31/-26 bp. Electrophoretic mobility shift assays with an anti-GATA-4 antibody confirmed that GATA-4 bound to these two sites with a dissociation constant of 1.56 nM (-79 site) and 0.65 nM (-31 site). Coexpression of GATA-4 stimulated EPHX1 promoter activity up to 7.5-fold in a dose-dependent manner. Endogenous EPHX1 message in HepG2 cells was also significantly increased by overexpression of GATA-4. Mutating the -79 element resulted in a 65% loss of promoter activity, while mutating the -31 element had no effect on basal activity but greatly reduced the response to additional GATA-4. In HeLa cells, which do not express GATA-4, EPHX1 activity was negligible; however, activity could be reconstituted by the addition of exogenous GATA-4. These results demonstrate that GATA-4 plays a critical role in regulating EPHX1 expression.

It is often desired to identify further homologs of a family of biological sequences from the ever-growing sequence databases. Profile hidden Markov models excel at capturing the common statistical features of a group of biological sequences. With these common features, we can search the biological database and find new homologous sequences. Most general profile hidden Markov model methods, however, treat the evolutionary relationships between the sequences in a homologous group in an ad-hoc manner. We hereby introduce a method to incorporate phylogenetic information directly into hidden Markov models, and demonstrate that the resulting model performs better than most of the current multiple sequence-based methods for finding distant homologs.

To examine 10 cases with primary cutaneous CD30-positive anaplastic large cell lymphoma (ALCL), analyze their clinical manifestations and pathological and immunohistochemical features, and improve early diagnosis of this disease.We studied the morphological characteristics of primary cutaneous CD30-positive ALCL using histopathological methods. Leukocyte common antigen (LCA), CD20, CD30, CD45RO, CD68, epithelial membrane antigen (EMA), cytokeratin (CK) and HMB45 antibodies were used to determine the expression of their respective antigens from routine paraffin samples of the patients.Ten patients (7 men and 3 women, aged 31 to 84 years) complained of subcutaneous masses or papular eruptions over their lower trunks and extremities. Histopathologically, the lesions were composed of numerous large round or oval pleomorphic cells. The cytoplasm was usually abundant, amphophilic or basophilic, and finely vacuolated. Nuclei were commonly eccentrically localized and lobated or horseshoed in shape, and multinucleated giant cells and Reed-Sternberg-like cells were seen. Nucleoli were generally multiple and large. Of the 10 patients, tumor cells displayed positive antigen expression of CD30 in all cases, positive CD45RO in 6 cases, positive CD20 in only 1 case, but negative CD45RO and CD20 expressions in 3 cases. Two patients died at 7 weeks and 3.4 years of follow-up, respectively.Our study highlights the importance of histopathologic features and positive CD30 staining for differentiation of this disease from other malignant skin tumors.

OBJECTIVE: To study the mechanism of inhibitory effect of Ubenimex on human leukemic cells. METHODS: K562 and HL60 cells were treated with Ubenimex at different concentrations, and the growth inhibition was analysed by MTT assay. Cell apoptosis was evaluated by light microscopy, agrose gel electrophoresis, TUNEL labeling method and flow cytometry (FCM) assay. RESULTS: (1)Treatment with Ubenimex remarkably inhibited the growth of HL60 cells, the IC(50) of Ubenimex for HL60 cells was 13.03mgr;g/ml. But K562 cells were less sensitive than HL60. Ubenimex inhibited the growth of HL60 and K562 cells in a dose-dependent manner. (2)Apoptosis of leukemic cells was induced by Ubenimex, which was shown by the changes in morphology, DNA ladder on agrose gel, TUNEL labeling,typical peak before G1 phase of cell cycle and the positive of Annexin V(FITC) on the cells membrane with FCM. (3)Ubenimex induced apoptosis of K562 and HL60 cells in a dose-and-time-dependent manner. (4)The cell cycle analysis by FCM showed that the HL60 cells were blocked in G1 phase after treated by Ubenimex. Conclution Ubenimex can efficiently induce apoptosis of HL60 and K562 cells, this may be one of the mechanisms for inhibiting effect of Ubenimex on leukemia.

The magnetic phase separation and exchange bias effect in off-stoichiometric Ni2Mn1.4Ga0.6 alloy were reported. Spontaneous exchange bias after zero-field cooling was observed in this alloy. The magnetic nature of ground state was characterized as phase separation with non-percolated ferromagnetic domains in spin glass matrix due to the spatial composition fluctuation and competing ferromagnetic and antiferromagnetic interactions. A mechanism of field-induced growth of ferromagnetic domains changing from non-percolating to percolating state was proposed for the appearance of spontaneous exchange bias.