INTRODUCTION: Centrifugal blood pumps have been widely adopted in conventional adult cardiopulmonary bypass and circulatory assist procedures. Different brands of centrifugal blood pumps incorporate distinct designs which affect pump performance. In this adult extracorporeal life support (ECLS) model, the performances of two brands of centrifugal blood pump (RotaFlow blood pump and CentriMag blood pump) were compared. METHODS: The simulated adult ECLS circuit used in this study included a centrifugal blood pump, Quadrox D membrane oxygenator and Sorin adult ECLS tubing package. A Sorin Cardiovascular(R) VVR(R) 4000i venous reservoir (Sorin S.p.A., Milan, Italy) with a Hoffman clamp served as a pseudo-patient. The circuit was primed with 900ml heparinized human packed red blood cells and 300ml lactated Ringer's solution (total volume 1200 ml, corrected hematocrit 40%). Trials were conducted at normothermia (36( degrees )C). Performance, including circuit pressure and flow rate, was measured for every setting analyzed. RESULTS: The shut-off pressure of the RotaFlow was higher than the CentriMag at all measurement points given the same rotation speed (p < 0.0001). The shut-off pressure differential between the two centrifugal blood pumps was significant and increased given higher rotation speeds (p < 0.0001). The RotaFlow blood pump has higher maximal flow rate (9.08 +/- 0.01L/min) compared with the CentriMag blood pump (8.37 +/- 0.02L/min) (p < 0.0001). The blood flow rate differential between the two pumps when measured at the same revolutions per minute (RPM) ranged from 1.64L/min to 1.73L/min. CONCLUSIONS: The results obtained in this experiment demonstrate that the RotaFlow has a higher shut-off pressure (less retrograde flow) and maximal blood flow rate than the CentriMag blood pump. Findings support the conclusion that the RotaFlow disposable pump head has a better mechanical performance than the CentriMag. In addition, the RotaFlow disposable pump is 20-30 times less expensive than the CentriMag.

Screening biological readouts in cell culture are increasing in frequency and throughput. In such assays, cell types may be rare and reagents or compounds may be expensive often resulting in a reduced number of conditions and/or replicates. "Tubeless" microfluidics offers a method to reduce this burden, as has been previously shown.1 In addition the In Cell Western (ICW) has recently been adapted to microfluidic cultures allowing high throughput analysis of immunocytochemistry (ICC) in microfluidic channels.2 Combining automated liquid handling in tubeless microfluidics with the ICW provides rapid and quantitative high throughput cell-based screens. Here we validate this platform using three parameters: operational robustness (pipetting reliability), cell seeding consistency, and cell staining consistency (both nuclear and antibody). Integration of liquid handling with microfluidics was found to be over 97% operationally robust. Cell seeding consistency between each microchannel and within each microchannel was found to be within a standard deviation of less than 5% and 6% respectively. Finally, through optimization of liquid handling steps, uniformity between all the channels was found for both nuclear and antibody staining. These results lay the foundation to perform most standard ICW assays using automated tubeless microfluidics.

Gold nanoparticles (AuNPs) are widely used as colorimetric probes for biosensing, relying on their unique particle size-dependent and/or interparticle distance-dependent extinction spectrum and solution color. Herein, we describe an AuNP-based colorimetric assay to detect binding interactions between nuclear hormone receptors and their corresponding DNA-binding elements, particularly the human estrogen receptors (ERalpha and ERbeta) and their cognate estrogen response elements (EREs). We found that the protein-DNA (ER-ERE) complexes can stabilize citrate anion-capped AuNPs against salt-induced aggregation to a larger extent than the protein (ER) or the DNA (ERE) alone, due to their unique molecular size and charge properties that provide a strong electrosteric protection. Moreover, our results show that the extent of stabilization is sequence-dependent and can distinguish a single base variation in the ERE associated with minor changes in protein-DNA binding affinity. With this assay, many important parameters of protein-DNA binding events (e.g., sequence selectivity, distinct DNA binding properties of protein subtypes, binding stoichiometry, and sequence-independent transient binding) can be determined instantly without using labels, tedious sample preparations, and sophisticated instrumentation. These benefits, in particular the high-throughput potential, could enable this assay to become the assay of choice to complement conventional techniques for large scale characterization of protein-DNA interactions, a key aspect in biological research.

PURPOSE: This cohort study evaluated the factors associated with discontinuation of antihypertensive pharmacotherapy among Chinese patients who were prescribed beta-blockers. We tested the hypothesis that patients' age, gender, socioeconomic status, clinical settings and the number of comorbidities were significantly associated with the discontinuation of beta-blockers. METHODS: From a validated clinical database we included all adult patients 18 years or older who were prescribed a beta-blocker in any government primary care clinic in one large territory of Hong Kong during 01 January 2004-30 June 2007. We evaluated the cumulative incidence of drug discontinuation within 180 days of the prescriptions and the factors associated with discontinuation of beta-blockers by multivariable regression analysis. RESULTS: From a total of 19,177 eligible patients (mean age = 59.1 years), 20.8% discontinued their medication. Younger patients [aged <50 years; adjusted odds ratios (aOR) 0.41-0.52 for patients aged >/=50 years; p < 0.001], female gender (aOR 0.87 for males, p = 0.001), fee-waivers (aOR 0.78 for fee-payers, p < 0.001), attendances in family medicine specialist clinics (FMSC) (aOR 1.49, p < 0.001) and staff clinics (aOR 2.32, p < 0.001), residence in more urbanized areas (aOR 0.80 for North District, p < 0.001), new visits (aOR 0.55 for follow-up visits, p < 0.001) and absence of concomitant comorbidities (aOR 0.60 for one comorbidity, p < 0.001; aOR 0.56 for two comorbidities, p = 0.002) were positively associated with drug discontinuation. CONCLUSIONS: Patients who were prescribed beta-blockers with these associated factors should be monitored more closely for antihypertensive drug adherence.

Quantum dots (QDs), also named semiconductor nanocrystals, have initiated a new realm of bioscience by combining nanomaterials with biology, which will profoundly influence future biological and biomedical research. In this review, we describe the extraordinary optical properties of QDs and developments in methods for their synthesis. We focus on fluorescent imaging with QDs both in vitro and in vivo, and the cytotoxicity of QDs and potential barriers to their use in practical biomedical applications. Finally, we provide insights into improvements aimed at decreasing the cytotoxicity of QDs and the future outlook of QD applications in biomedical fields.

The InGaZnO thin films are fabricated on the quartz glass using pulsed laser deposition (PLD), where the target is prepared by mixing the Ga(2)O(3), In(2)O(3), and ZnO powders at a mol ratio of 1:1:8 before the solid-state reactions in a tube furnace at the atmospheric pressure. The product thin films were characterized comprehensively by X-ray diffraction, atomic force microscopy, Hall-effect investigation, and X-ray photoelectron spectroscopy. Thus, we demonstrate semiconductor thin-film materials with high smoothness, high transmittance in visible region, and excellent electrical properties.

The mechanisms controlling organismal aging have yet to be clearly defined. In our recent paper [1], we revealed thatTAp63, the p53 family member, is a critical gene in preventing organismal aging by controlling the maintenance of dermal and epidermal precursor and stem cells critical for wound healing and hair growth. In the absence of TAp63, dermal stem cells (skin-derived precursors or SKPs) in young mice are hyperproliferative. As early as one month of age, SKPs and epidermal precursor cells exhibit signs of premature aging including a marked increase in senescence, DNA damage, and genomic instability resulting in an exhaustion of these cells and an overall acceleration in aging. Here, we discuss our findings and its relevance to longevity, regenerative medicine, and tumorigenesis.

The clinical characteristics and cerebrospinal fluid (CSF) cytological features of 34 hospitalized patients with leptomeningeal carcinomatosis (LC) were studied. Most patients presented with signs of meningeal irritation (19 cases) and intra-cranial hypertension (23 cases). Computed tomography (CT) and/or magnetic resonance imaging (MRI) revealed brain parenchymal lesions, hydrocephalus and leptomeningeal enhancement (nine, six and eight cases, respectively). The CSF changes included high opening pressure (21 cases), increased white blood cell count (23 cases), elevated protein levels (25 cases) and low glucose levels (17 cases). Malignant cells were found in all CSF specimens and 32 cases had malignant cells in their initial CSF examinations. High serum levels of carcinoembryonic antigen (CEA) occurred in 11 patients. Signs of meningeal irritation and intra-cranial hypertension were common. It is concluded that serum CEA measurement along with CT and MRI scanning are helpful in the diagnosis of LC. Crucially, however, CSF cytology could be the most important technique for the diagnosis of LC.

ABSTRACT: BACKGROUND: DNA-methyltransferase (DNMT)-3A plays an important role in the development of embryogenesis and the generation of aberrant methylation in carcinogenesis. The aim of this study was to investigate the role of a DNMT3A promoter genetic variant on its transcriptional activity and to evaluate the association between DNMT3A gene polymorphism and the susceptibility to gastric cancer (GC) and oesophagus carcinoma (EC) in the Chinese population. METHODS: We selected one of the single nucleotide polymorphisms (SNPs) -448A>G in the DNMT3A promoter region and evaluated its effect on activity using a luciferase assay. -448A>G polymorphisms of DNMT3A were determined by polymerase chain reaction / restriction fragment length polymorphism and confirmed by sequencing. The distribution of -448A>G polymorphisms was detected in 208 GC patients and 346 healthy controls matched for age and gender. The distribution of -448A>G polymorphisms was also detected in 96 EC patients and matched 241 healthy controls. The association of -448A>G polymorphisms of DNMT3A and the risk of GC and EC was evaluated by stratified analysis according to the patient's age and gender. RESULTS: In a promoter assay, carriage of the -448 A allele showed a significantly higher promoter activity ( > two fold) compared with the -448G allele (P<0.001). The allele frequency of -448A among GC patients and controls was 32.9% versus 19.9%, respectively. Overall, we found that, compared with GG carriers, the DNMT3A -448AA homozygotes has a > six fold increased risk of GC. Stratification analysis showed that AA homozygotes have a more profound risk in the subgroups of individuals at the age range [less than or equal to] 60 years in GC. However, individuals with -448AG and -448AA were not statistically significantly associated with an increased risk of EC compared with those carried the -448GG genotype. CONCLUSIONS: The DNMT3A -448A>G polymorphism is a novel functional SNP and contributes to its genetic susceptibility to GC. -448A>G can be used as a stratification marker to predict an individual's susceptibility to GC, especially in the subgroups of individuals at the age range [less than or equal to] 60 years. However, the relative distribution of -448A>G in EC can not be used as a prediction marker in order to evaluate an individual's susceptibility to EC.

The aim of this study was to detect the expression pattern of DNA methyltransferase 3B (DNMT3B) variants in primary gastric cancer (GC) and to explore the clinical significance of DNMT3B variants in gastric carcinogenesis. Specific polymerase chain reaction (PCR) primer sets were designed to distinguish individual DNMT3B variants according to their splicing patterns. Expression levels of DNMT3B variants were assessed by quantitative real-time RT-PCR in gastric cancer tissue, normal gastric mucosae and GC cell lines. The relationship between the expression patterns of the DNMT3B variants and corresponding clinical information was analyzed by observing the expression levels of different variants in the tumors. These results demonstrate that DNMT3B overexpression is related to late phase invasion (P=0.029) and intestinal type (P=0.012) in GC. DNMT3B3 expression was higher in normal tissue, compared to tumor tissue (P=0.033). In contrast, only 18, 32 and 35% of the patient tumors overexpressed DNMT3B1, DNMT3B4 and DNMT3B5, respectively. While taking into account environmental factors (H. pylori, Epstein-Barr virus infection), H. pylori infection elevated DNMT3B1 and DNMT3B3 variants in tumors, while increasing DNMT3B4 in both tumor and non-cancerous tissues. Our findings indicated that the expression of DNMT3B3 is the major splice variant in normal gastric mucosae and may be affected by H. pylori infection. Elevated DNMT3B variants may influence the progression of gastric cancer and may possibly be a powerful indicator for the disease.

ADP-ribosylation factor 6 (Arf6) is a small GTPase which influences membrane receptor trafficking and the actin cytoskeleton. In adipocytes, Arf6 regulates the trafficking of the glucose transporter, GLUT4 and consequently insulin-stimulated glucose transport. Previous studies also indicated a role of Arf6 in adrenergic receptor trafficking but whether this contributes to the control of lipolysis in adipocytes remains unknown. This was examined in the present study using RNA interference (RNAi) and pharmaceutical inhibition in murine cultured 3T3-L1 adipocytes. Down regulation of Arf6 by RNAi impairs isoproterenol-stimulated lipolysis specifically but does not alter triacylglycerol (TAG) synthesis or the insulin signaling pathway. Neither total TAG amounts nor TAG fatty acid compositions are altered. The inhibitory effect on lipolysis is mimicked by dynasore, a specific inhibitor for dynamin, which is required for endocytosis. In contrast, lipolysis triggered by reagents that bypass events at the plasma membrane (e.g. forskolin, isobutylmethylxanthine (IBMX) or 8-Bromo-cAMP) is not affected. Moreover, Arf6 protein levels in white adipose tissues are markedly increased in ob/ob mice, while they are decreased in obesity resistant CD36 null mice. These changes reflect at least in part alterations in Arf6 mRNA levels. Collectively, these results suggest a role of the endocytic pathway and its regulation by Arf6 in adrenergic stimulation of lipolysis in adipocytes and potentially in the development of obesity.

The present study aims to clarify the inhibitive effect of the compounds from Rheum palmatum L. on the SARS-3CL protease. The SARS-CoV 3CL gene was amplified from RNA of the SARS virus by PCR. The SARS-CoV 3CL protease was purified from a colon bacillus recombinant. Drugs and 3CL protease were incubated together. The inhibition rate and IC₅₀ were calculated based on absorbance. Components from the Rheum palmatum L. had a high level of anti-SARS-CoV 3CL protease activity. The IC₅₀ was 13.76 +/- 0.03 mug/mL and the inhibition rate was up to 96%. In conclusion, extracts from Rheum palmatum L. have a high level of inhibitory activity against 3CL protease, suggesting that extracts from Rheum palmatum L. may represent a potential therapeutic for SARS.

The chiral tris-monodentate imidazolinyl ligands 1 a-c exhibit a strong tendency to form the discrete, helical [2+3] nanocages 3 ([1(2)2(3)]) with tartaric acids 2. Circular dichroism (CD) spectra and theoretical calculations reveal that supramolecular handedness of capsulelike architectures is determined only by the chirality of the imidazolinyl ligands rather than tartaric acids. The chirality of imidazolinyl ligands is transferred to the helicity of the complexes through the directed hydrogen bonds between the N3 atom of imidazoline rings and the carboxyl of tartaric acids. These hydrogen-bonded nanocages can spontaneously self-assemble into spherical vesicles, during which the hydrogen bonding that arises from the hydroxyl groups of tartaric acids plays a crucial issue. The vesicles formed by [{(S,S,S)-1 a}(2)(2(L))(3)] (3 a) may further evolve into microspheres that gelate organic solvents after being aged at -20 degrees C for 24 h, and can also be unprecedentedly transformed to tubular assemblies capable of rigidifying the solvents when subjected to ultrasound irradiation.

A type of 1,1'-binaphthyl-based imidazolium chemosensor module has been synthesized for the highly selective recognition of tryptophan (Trp) among the eleven alpha-amino acids investigated in aqueous solutions via synergistic effects of multiple hydrogen bonding and electrostatic interactions. These results have demonstrated that the C-2 hydrogen atom of the imidazolium ring plays a key role as a hydrogen bond donor. The UV/vis, fluorescence and mass spectral studies have indicated that a 1 : 1 complex is formed between the host and tryptophan. The binding affinity and selectivity of the cleft-like receptor (R)- with l-Trp are superior to those of (R)-. In spite of an inferior selectivity towards various aromatic amino acids, the macrocyclic (R)- displays a remarkable enantiodiscrimination for the two enantiomers of tryptophan with a K(D)/K(L) value as high as 6.2.

In this study the gene encoding HSP70 was isolated from Phascoloma esculenta by homologous cloning and rapid amplification of cDNA ends (RACE). The full-length of cDNA (2520 bp) consists of a 5'-terminal untranslated region (UTR) (125 bp), a 3'-terminal UTR (421bp) with a canonical polyadenylation signal sequence (AATAAA), a poly (A) tail, and an open reading frame (ORF) (1974bp). The predicted molecular mass and isoelectric point for HSP70 is 71.6 kDa and 5.15, respectively. BLAST analysis showed that P. esculenta HSP70 gene shared high similarity. Classical HSP signature motifs, ATP/GTP-Binding Site Motif A, Bipartite Nuclear Targeting Sequence, the cytosolic HSP70 could be expressed in E. coli BL21. After purification, the recombinant pET-HSP70 protein was used to produce the polyclonal antibody in mice and the specificity of the antibody was confirmed by Western blot analysis. Fluorescent real-time quantitative PCR analysis showed that expression of Hsp70 in sipuncula was increased significantly after exposure to 10mM Zn for12h, Cd for 24h, Cu for 48h, and was exposure to 37 degrees C for 24h sea water.

A two-dimensional continuous wavelet transform employing a real mother wavelet is applied to phase analysis of spatial carrier fringe patterns. In this method, a Hilbert transform is first performed on a carrier fringe pattern to get an analytic signal. Then a two-dimensional wavelet transform is calculated for the signal that is yielded by the first transform. Finally, the height-demodulated phase information can be gotten from the wavelet transform coefficients at the wavelet ridge position. The performance of the proposed method has been evaluated by using computer-generated and real fringe patterns. The result performed better than that of one-dimensional real wavelet transform algorithms in the area with phase discontinuous points and high phase variation, especially when there is much noise in the fringe patterns. Computer simulations and experiments verified the validity of the proposed method.

Although Th17 cells play critical roles in the pathogenesis of many inflammatory and autoimmune diseases, their prevalence among tumor-infiltrating lymphocytes (TILs) and function in human tumor immunity remains largely unknown. We have recently demonstrated high percentages of Th17 cells in TILs from ovarian cancer patients, but the mechanisms of accumulation of these Th17 cells in the tumor microenvironment are still unclear. In this study, we further showed elevated Th17 cell populations in the TILs obtained from melanoma and breast and colon cancers, suggesting that development of tumor-infiltrating CD4(+) Th17 cells may be a general feature in cancer patients. We then demonstrated that tumor microenvironmental RANTES and MCP-1 secreted by tumor cells and tumor-derived fibroblasts mediate the recruitment of Th17 cells. In addition to their recruitment, we found that tumor cells and tumor-derived fibroblasts produce a proinflammatory cytokine milieu as well as provide cell-cell contact engagement that facilitates the generation and expansion of Th17 cells. We also showed that inflammatory TLR and nucleotide oligomerization binding domain 2 signaling promote the attraction and generation of Th17 cells induced by tumor cells and tumor-derived fibroblasts. These results identify Th17 cells as an important component of human TILs, demonstrate mechanisms involved in the recruitment and regulation of Th17 cells in tumor microenvironments, and provide new insights relevant for the development of novel cancer immunotherapeutic approaches.

There is no effective systemic therapy for patients with advanced hepatocellular carcinoma (HCC) except liver transplantation. Sorafenib, a multikinase inhibitor, has been shown to significantly increase overall survival (OS) in a randomized, placebo-controlled, phase III trial of patients with HCC (SHARP). The aim of this study was to evaluate the effectiveness of sorafenib for advanced HCC by carrying out a meta-analysis of randomized controlled trials that compared sorafenib-based therapy with other agent-based therapy. Randomized controlled trials comparing sorafenib or combined chemotherapy with placebo or combined chemotherapy in advanced HCC between 2000 and 2008 were identified and the data were extracted from reports. Outcomes analyzed were objective response rate, time to progression (TTP), OS, and toxicity. The summary hazard ratios (HRs), odds ratios, and their 95% confidence intervals (CIs) for mortality, objective response rate, and toxicity were estimated. All statistical tests were two-sided. Three trials including 924 patients were identified. Sorafenib-based chemotherapy was also associated with a 79% prolongation of TPP (HR = 0.58, 95% CI = 0.49-0.69, P<0.001), and a 37.3% increase in OS (HR = 0.66, 95% CI = 0.55-0.78, P<0.001). Despite significant increases in the frequencies of hand-foot syndrome and diarrhea in patients receiving sorafenib-containing chemotherapy, no significant difference in other toxic events was observed. This meta-analysis suggests that sorafenib-based chemotherapy is superior to placebo-based chemotherapy in terms of TPP and OS without increase in severe toxic effects.

In this paper, the interaction of DNA molecules with aqueous CdTe quantum dots (CdTe QDs), CdTe/SiO(2) composite nanoparticles (CdTe/SiO(2) NPs), and Mn-doped ZnSe quantum dots (Mn:ZnSe d-dots) was studied with ethidium bromide as a probe. The purpose of this work was to study the damage of DNA molecules induced by these three kinds of water-soluble nanoparticles. It was found that ionic strength, pH value and UV irradiation influenced the PL emission properties of CdTe QDs, CdTe/SiO(2) NPs and Mn:ZnSe d-dots, and also influenced the interaction of DNA molecules with them. Among the three kinds of nanoparticles, DNA molecules were most easily damaged by CdTe QDs whether in the dark or under UV irradiation. The CdTe/SiO(2) NPs led to much less DNA damage when compared with CdTe QDs, as a silica overcoating layer could isolate the QDs from the external environment. Mn:ZnSe d-dots as a new class of non-cadmium doped QDs demonstrated almost no damage for DNA molecules, which have great potentials as fluorescent labels in the applications of biomedical assays, imaging of cells and tissues, even in vivo investigations.

Chitosan is known to inhibit microorganisms of concern to plants, animals, and humans. However, the effect of chitosan on human enteric viruses of public health concern has not been extensively investigated. The purpose of this study was to determine the effect of chitosan on three human enteric viral surrogates: murine norovirus 1 (MNV-1), feline calicivirus F-9 (FCV-F9), and (ssRNA) bacteriophage MS2 (MS2). Chitosan oligosaccharide lactate (molecular weight of 5,000) and water-soluble chitosan (molecular weight of 53,000) at concentrations of 1.4, 0.7, and 0.35% were incubated at 37 degrees C for 3 h with equal volumes of each virus at high ( approximately 7 log PFU/ml) and low ( approximately 5 log PFU/ml) titers. Chitosan effects on each treated virus were evaluated with standardized plaque assays in comparison to untreated virus controls. The water-soluble chitosan at 0.7% decreased the FCV-F9 titer by approximately 2.83 log PFU/ml, with decreasing effects at lower concentrations, and also decreased MS2 at high titers by approximately 1.18 to 1.41 log PFU/ml, regardless of the concentration used. Chitosan treatments at the concentrations studied had no effect on MNV-1 at high titers. Chitosan oligosaccharide showed similar trends against the viruses, but to a lesser extent compared with that of water-soluble chitosan. When lower virus titers ( approximately 5 log PFU/ml) were used, plaque reduction was observed for FCV-F9 and MS2, but not MNV-1. The use of higher-molecular-weight chitosan and at higher concentrations with longer incubation may be necessary to inactivate MNV-1. These results in the plaque reduction of human enteric virus surrogates by chitosan treatment show promise for its potential application in the food environment.