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Published emission factors (EFs) often vary significantly, leading to high uncertainties in emission estimations. There are few reliable EFs from field measurements of residential wood combustion in China. In this study, 17 wood fuels and one bamboo were combusted in a typical residential stove in rural China to measure realistic EFs of particulate matter (PM), organic carbon (OC), and elemental carbon (EC), as well as to investigate the influence of fuel properties and combustion conditions on the EFs. Measured EFs of PM, OC, and EC (EF(PM), EF(OC), and EF(EC), respectively) were in the range of 0.38-6.4, 0.024-3.0, and 0.039-3.9 g/kg (dry basis), with means and standard derivation of 2.2 ± 1.2, 0.62 ± 0.64, and 0.83 ± 0.69 g/kg, respectively. Shrubby biomass combustion produced higher EFs than tree woods, and both species had lower EFs than those of indoor crop residue burning (p < 0.05). Significant correlations between EF(PM), EF(OC), and EF(EC) were expected. By using a nine-stage cascade impactor, it was shown that size distributions of PM emitted from tree biomass combustions were unimodal with peaks at a diameter less than 0.4 μm (PM(0.4)), much finer than the PM from indoor crop residue burning. Approximately 79.4% of the total PM from tree wood combustion was PM with a diameter less than 2.1 μm (PM(2.1)). PM size distributions for shrubby biomasses were slightly different from those for tree fuels. On the basis of the measured EFs, total emissions of PM, OC, and EC from residential wood combustion in rural China in 2007 were estimated at about 303, 75.7, and 92.0 Gg.

For comparison of arbitrary frequency signals, the paper proposed two levels of length vernier based on the time-space relationship are used in three levels of phase coincidence detecting circuits to extract the phase coincidence information by proper logic calculation. The length∕phase of each vernier is respectively corresponding to the accuracy and the resolution of detecting circuit. The time-space relationship is based on high-stability, high-accuracy, and high-speed of signal transmission. The method is effective to reduce the fuzzy region in the phase coincidence information and reach a higher measuring precision.

Aim: FTO is the most important polygene for obesity and type 2 diabetes. Our aims were to investigate whether a variant in FTO affects glucose dysregulation through its effect on fat accumulation or distribution, and when and how FTO influences fat accumulation and distribution and glucose dysregulation from birth until midlife., Methods: A total of 454 Japanese female university students (mean age: 20 years) and 132 middle-aged women (mean age: 50 years) who were the biological mothers of the students underwent the following: genotyping for rs1558902 in the FTO gene, assessment of fat accumulation and distribution, obesity-related metabolic traits and serum adipokine measurement. A subsample of 364 students reported their weight history since birth., Results: The A allele in rs1558902 was substantially less common in young and middle-aged Japanese women (18 and 17%, respectively) than in the European population (45%). The strong effect of genotype AA on BMI was evident at age 12, 15, 20 and 50 years whereas there was no effect on birth weight. In young and middle-aged women, the variant was strongly associated with higher body weight and fat mass. The effects on abdominal girth, fasting glucose, homeostasis model assessment insulin resistance and HbA1c were evident in mothers but not in students. In addition, genotype AA was associated with increased white blood cell counts and hsCRP in mothers only. Associations with fasting glucose, insulin resistance, and white blood cell counts remained after correction for BMI, abdominal girth and fat mass. In multiple logistic regression analysis, AA homozygote in FTO was associated with higher odds of overweight (BMI ≥25 kg/m(2)) in young (OR 1.73 (95%CI 1.06-30.0)) and middle-aged women (OR 1.73 (95%CI 1.06-30.0)). It was also associated with higher odds of abdominal fat accumulation (abdominal girth ≥90 cm, OR 1.73 (95%CI 1.06-30.0)) and fasting hyperglycemia (≥100 mg/dL) (OR 1.87(95%CI 1.05-40.4)) in middle-aged mothers., Conclusion: Despite the small sample size, the low average BMI, and the low risk allele frequency, a genetic variation at the FTO locus was related to greater weight gain before age 12 in Japanese women. At age 20, it was related to general adiposity. In midlife, however, it was related to abdominal adiposity in addition to general adiposity, fasting hyperglycemia, higher HbA1c and subtle systemic inflammation. Fasting hyperglycemia associated with higher HbA1c in midlife was independent of its effects on general and abdominal adiposity.

HdeA protein is a small, ATP-independent, acid stress chaperone that undergoes a dimer-to-monomer transition in acidic environments. The HdeA monomer binds a broad range of proteins to prevent their acid-induced aggregation. To understand better HdeA's function and mechanism, we perform constant-pH molecular dynamics simulations (CPHMD) to elucidate the details of the HdeA dimer dissociation process. First the pK(a) values of all the acidic titratable groups in HdeA are obtained and reveal a large pK(a) shift only for Glu(37). However, the pH-dependent monomer charge exhibits a large shift from -4 at pH > 6 to +6 at pH = 2.5, suggesting that the dramatic change in charge on each monomer may drive dissociation. By combining the CPHMD approach with umbrella sampling, we demonstrate a significant stability decrease of the HdeA dimer when the environmental pH changes from 4.0 to 3.5 and identify the key acidic residue-lysine interactions responsible for the observed pH sensing in HdeA chaperon activity function., (© 2011 American Chemical Society)

Sophocarpine, one of the major alkaloid compounds isolated from Sophora pachycarpa, is highly valued and important in traditional Chinese medicine. In the present study, we aimed to explore the possible mechanisms underlying sophocarpine-mediated neuroprotection against transient focal cerebral ischemia. Sophocarpine (5, 10, or 20mg/kg) was given 30min before focal ischemia was induced in rats by occlusion of the middle cerebral artery. After sophocarpine treatment, the total infarct volume was significantly decreased in comparison to the ischemia-reperfusion values. The results of a neurological evaluation were significantly improved in the sophocarpine treated group when compared to controls. The number of TUNEL-positive cells was significantly reduced compared to the untreated ischemic group. Results of Western blotting and immunohistochemical staining indicated that pretreatment with sophocarpine down-regulated the expression of acid-sensing ion channel 1 (ASIC1) in the ischemic cortex. These results suggest that sophocarpine ameliorated the ischemic injury induced by transient focal cerebral ischemia in rats and that this neuroprotective effect might be related to the anti-ASIC1 channel and anti-apoptotic action of sophocarpine., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Cyclooxygenase-2 (COX-2), an inducible isoform of cyclooxygenase, has been reported to be correlated with tumorigenesis, tumor progression and metastasis. The present study was designed to investigate the clinicopathological and prognostic significance of COX-2 in Chinese patients with prostate cancer. Firstly, RT-PCR and Western blot assays were performed to detect the expression of COX-2 mRNA and protein in prostate cancer cell lines and 20 tissue samples (tumor or corresponding non-tumor). Next, immunohistochemistry was performed to detect the expression of COX-2 protein in 88 prostate cancer tissue samples. Finally, the correlation between COX-2 expression and clinicopathological factors and patient survival was evaluated. We found that the expression levels of COX-2 mRNA and protein showed significant difference among four prostate cancer cell lines. Moreover, the levels of COX-2 mRNA and protein were significantly higher in prostate cancer tissues than in corresponding non-tumor tissues. COX-2 staining was positive in the cytoplasm of prostate cancer cells. High-COX-2 expression was correlated with the Gleason score (P=0.009), tumor stage (P=0.012), and lymph-node status (P=0.036). Furthermore, patients with high-COX-2 expression showed lower disease-free (P=0.014) and overall survival (P=0.047) rates than those with low-COX-2 expression. Univariate and multivariate analyses suggested that the status of COX-2 protein expression was an independent prognostic indicator for patients' survival. Taken together, higher COX-2 protein expression might provide an independent prognostic marker for Chinese patients with prostate cancer who have undergone surgery., (Copyright © 2009 Elsevier GmbH. All rights reserved.)

Objective: To establish fluorescence resonance energy transfer (FRET) assay method of detecting proteolytic activity of non-structural protein 3-4A (NS3-4A) serine protease of hepatitis C virus (HCV) for high throughput screening inhibitors against HCV in vitro., Methods: HCV recombinant plasmid pMAL~c2/NS3-4A was transformed into the E.coli strain K12TB1. Maltose-binding-protein (MBP) NS3-4A fusion protein expression was induced by adding isopropyl-β-D-thiogalacto-pyranoside (IPTG) and purified by affinity chromatography. The proteolytic activity of MBP-NS3-4A protease was analyzed by FRET with the special protease substrate. The reaction system in this model was optimized, and the reliability of the model was evaluated., Results: High throughput screening model for HCV NS3-4A protease inhibitors was established, and the best concentrations of enzyme and substrate were optimized. In the model, the Km value of protease was 4.74 μmol/L, Z factor was up to 0.80, and coefficient of variation (CV) was 1.91%. BILN 2061, one of the known HCV protease inhibitors, was measured with the Ki of 0.30 nmol/L., Conclusion: The assay model using FRET method for HCV NS3 4A serine protease is stable and reliable, and the model is suitable for high throughput screening for HCV NS3 4A protease inhibitors.

Stringent air-quality control measures were implemented for the 2008 Beijing Olympic Games. This large-scale manmade experiment provided an opportunity to evaluate the effectiveness of measures to reduce the reactivity of hydrocarbons (HCs) from emission sources, which is important for ground-level ozone abatement. Photochemical initial concentrations (PICs), i.e., the levels of HCs from sources before undergoing chemical reactions, were calculated from ambient measurements. PICs obtained using the ratio method for HCs and the sequential reaction model for alkyl nitrates were in good agreement. Propene, 1-butene, iso-butene, trans-2-butene, cis-2-butene, trans-2-pentene, and m,p-xylene were identified as key reactive species in terms of their photochemical consumptions and correspondent ozone formation potentials (OFPs). During the Olympics and Paralympics, the PICs of these seven species were reduced by 27-66%, contributing 20% to the reduction in total PICs and 60% to the reduction in total OFP compared with June levels. Source apportionments from the chemical mass balance model indicated that gasoline vehicle exhaust was the predominant contributor to the key reactive species (45-78%). Reductions of gasoline vehicle exhaust during the Olympics and Paralympics explained 53-77% and 59-68% of the reductions in PICs of the key reactive HCs and total OFP, respectively.

Aim and Methods: We assessed the relationship of the body mass index (BMI) of 187 college female students aged 18 years with the reported BMI of their middle-aged biological parents measured on 2 occasions: when the parents were 18-20 years old and at the time of the study. The relationships of fat mass measured using whole body dual energy X-ray absorptiometry (DXA) and serum leptin levels were also determined between 148 daughters and middle-aged parents (148 mothers and 59 fathers)., Results: The BMI of daughters was associated with their mothers' BMI (r=0.30, p<0.0001) but not with their fathers' BMI measured when they were 18 years old. Daughters' BMI showed a stronger association with the current BMI of their mothers BMI (r=0.36, p<0.0001) than that of their fathers' BMI (r=0.19, p=0.01). In addition, the serum leptin levels of daughters were correlated with their mothers' leptin values (r=0.22, p=0.04). Further, not only total body fat mass (r=0.19, p<0.05) but also fat mass in the trunk (r=0.18, p<0.05) and legs (r=0.17, p<0.05) was associated between daughters and their mothers., Conclusion: The significant correlation between daughters' and mothers' BMI measured when their mothers were 18 years old did not result from shared environmental factors, including the intrauterine environment. The results in the present study therefore suggest that adiposity in 18-year-old daughters may be influenced by the maternal effect. The associations of serum leptin and DXA-derived fat mass between daughters and their mothers may support our hypothesis.

We extend the weighted ensemble (WE) path sampling method to perform rigorous statistical sampling for systems at steady state. A straightforward steady-state implementation of WE is directly practical for simple landscapes, but not when significant metastable intermediates states are present. We therefore develop an enhanced WE scheme, building on existing ideas, which accelerates attainment of steady state in complex systems. We apply both WE approaches to several model systems, confirming their correctness and efficiency by comparison with brute-force results. The enhanced version is significantly faster than the brute force and straightforward WE for systems with WE bins that accurately reflect the reaction coordinate(s). The new WE methods can also be applied to equilibrium sampling, since equilibrium is a steady state.

In this article, the propagation of guided thermoelastic waves in the circumferential direction of orthotropic cylindrical curved plates subjected to stress-free, isothermal boundary conditions is investigated in the context of the Green-Naghdi (GN) generalized thermoelastic theory (without energy dissipation). The coupled wave equations and heat conduction equation are solved by the Legendre orthogonal polynomial series expansion approach. The convergency of the method is discussed through a numerical example. The dispersion curves of thermal modes and elastic modes are illustrated simultaneously. Dispersion curves of the corresponding pure elastic cylindrical plate are also shown to analyze the influence of the thermoelasticity on elastic modes. The displacement, temperature and stress distributions are shown to discuss the differences between the elastic modes and thermal modes. A thermoelastic cylindrical plate with a different ratio of radius to thickness is considered to discuss the influence of the ratio on the characteristics of circumferential thermoelastic waves., (2009 Elsevier B.V. All rights reserved.)

Aim: To investigate the feasibility of using diffusion-weighted MR imaging (DWI) to accurately measure breast cancer extension., Materials and Methods: The extensions of 59 breast lesions were investigated on DW images and apparent diffusion coefficient (ADC) maps and compared to the pathological exams., Results: Three groups were observed: accurate, overdiagnosis, and false negative. There were no significant differences seen in accurate or false-negative group when b was 500 and 1000 s/mm(2) when two lesions in the overdiagnosis group at both b values., Conclusion: DWI and the ADC value have potential for evaluating cancer extension., (Copyright 2010 Elsevier Inc. All rights reserved.)

The "weighted ensemble" method, introduced by Huber and Kim [Biophys. J. 70, 97 (1996)], is one of a handful of rigorous approaches to path sampling of rare events. Expanding earlier discussions, we show that the technique is statistically exact for a wide class of Markovian and non-Markovian dynamics. The derivation is based on standard path-integral (path probability) ideas, but recasts the weighted-ensemble approach as simple "resampling" in path space. Similar reasoning indicates that arbitrary nonstatic binning procedures, which merely guide the resampling process, are also valid. Numerical examples confirm the claims, including the use of bins which can adaptively find the target state in a simple model.

Trichophyton rubrum (T.rubrum) is one of the most common human fungal pathogens that cause chronic infections of the skin and nails. To identify antifungal responsive genes, cDNA microarray analysis was performed for T. rubrum to reveal global transcriptional profiles of drug-specific responses to 5-Flucytosine (5-FC). cDNA microarray was constructed from the T. rubrum expressed sequence tag (ESTs) database, the minimum inhibitory concentration (MIC) of 5-FC was determined, and microarray hybridization and data analysis were applied. The expression pattern of 7 genes observed by microarray was confirmed by the quantitative real-time reverser transcription polymerase chain reaction (RT-PCR). Data analysis indicated that a total of 474 genes were found differentially expressed, 196 showed an increase in expression and 278 showed a decrease in expression. Marked down-regulation of genes involved in nucleotide metabolism (such as CDC21), transcription (such as E2F1), and RNA processing (such as SGN1, RIM4 and NOP1) was observed. Other genes involved in signal transduction, chaperones, inorganic ion transport, secondary metabolite biosynthesis, amino acid transport, lipid transport and potential drug resistance mechanism were also affected by 5-FC. Quantitative real-time RT-PCR of the selected genes confirmed the reliability of the microarray results. This is the first analysis of transcriptional profiles in response to 5-FC for T. rubrum. The findings may be valuable for the identification of genes involved in mechanisms of action and mechanisms of antifungal drug resistance of 5-FC.

KiSS-1 has been identified as a putative metastasis-suppressor gene in human melanomas and breast cancer cell lines. Although loss of KiSS-1 expression has been associated with progression and poor prognosis of various cancers, the exact role of KiSS-1 expression in HCC is not well-defined. Our study investigated KiSS-1 expression levels in HCC and its role in invasion and metastasis of human HCC. The expression levels of KiSS-1 and MMP-9 protein were determined by tissue microarray (TMA) serial sections, immunohistochemistry and semi-quantitative image analysis. All clinical and histological data obtained were subjected to statistical analysis. The expression of KiSS-1 protein in HCC and intrahepatic metastasis lesions was significantly lower (P < 0.01) when compared with non-tumor liver tissue and normal liver tissue. Multivariate analysis revealed a significant inverse correlation between KiSS-1 expression and o1 TNM stage, (F = 7.113, P < 0.01) and o2 intrahepatic metastasis (t = 2.898, P < 0.01). Loss of KiSS-1 in intrahepatic metastasis versus primary carcinomas was statistically significant (P<0.01). We also found a negative correlation between KiSS-1 and MMP-9 expression in HCC (r = -0.506, P < 0.01). We conclude that loss of KiSS-1 during HCC metastasis, along with a concomitant upregulation of MMP-9 suggests a possible mechanism for cell motility and invasion during HCC metastasis, with KiSS-1 emerging as a possible therapeutic target during HCC metastasis., ((c) 2009 Wiley-Liss, Inc.)

Mutants of a highly pathogenic, porcine reproductive, and respiratory syndrome virus (PRRSV), JXA1 strain, were prepared by continuous in vitro passage. Genomic sequence comparisons were made between mutants obtained at different passages and the parental strain JXA1. The mutant strain obtained at passage 80 contained a 12 nucleotide insertion and 108 nucleotide mutations that resulted in 45 amino acid changes. Most of these changes (89%) occurred between passage 10 and 45 and were genetically stable for the next 35-70 passages. A comparison of the mutants, their parental strain, and several American PRRSV strains, identified 13 characteristic amino acid changes. These sites, as well as the distinct 12 nucleotide insertion, represent possible genetic markers for the evaluation of live vaccine applications, particularly for additional studies of the safety and potency of live PRRSV vaccines.

Human STAU1 is one member of the family of double-stranded RNA (dsRNA)-binding proteins. It is thought to function in transporting mRNA, controlling translation and eliciting mRNA decay in neurons, and to function in infection of influenza virus and human immunodeficiency virus type 1 (HIV-1). Four transcripts coding two isoforms have been identified before. In this study, we have isolated a novel transcript of STAU1, coding a novel isoform that has six amino acids more (SFPLKQ) than isoform a. In order to examine the tissue distribution of this novel isoform, we have performed RT-PCR experiments and the analysis showed that it was highly expressed in heart, liver, kidney and pancreas.

The computational sampling of rare, large-scale, conformational transitions in proteins is a well appreciated challenge-for which a number of potentially efficient path-sampling methodologies have been proposed. Here, we study a large-scale transition in a united-residue model of calmodulin using the "weighted ensemble" (WE) approach of Huber and Kim. Because of the model's relative simplicity, we are able to compare our results with brute-force simulations. The comparison indicates that the WE approach quantitatively reproduces the brute-force results, as assessed by considering (i) the reaction rate, (ii) the distribution of event durations, and (iii) structural distributions describing the heterogeneity of the paths. Importantly, the WE method is readily applied to more chemically accurate models, and by studying a series of lower temperatures, our results suggest that the WE method can increase efficiency by orders of magnitude in more challenging systems.

Objective: To explore the clinical features and treatment results of brachial plexus palsy caused by halo traction before posterior correction in patients with severe scoliosis., Methods: A total of 300 cases of severe scoliosis received halo traction before posterior correction in our department from July 1997 to November 2004. Among them, 7 cases were complicated with brachial plexus palsy. The average Cobb angle was 110 degree (range, 90 degree-135 degree). Diagnoses were made as idiopathic scoliosis in 1 case, congenital scoliosis in 3 cases, and neuromuscular scoliosis in 3 cases. Additionally, diastematomyelia and tethered cord syndrome were found in 3 cases and thoracolumbar kyphosis in 2 cases. Weight of traction was immediately reduced when the patient developed any abnormal neurological symptoms in the upper extremity, and rehabilitation training was undertaken. Simultaneously, neurotrophic pharmacotherapy was applied, and the neurological function restoration of the upper limbs and the recovery time were documented., Results: Traction was used for an average of 3.5 weeks (range, 2-6 weeks) before spinal fusion for these 7 patients. The average traction weight was 8 kg, which was 19% on average (range, 13%-26%) of the average body weight (40.2 kg). These 7 patients had long and thin body configuration with a mean height of 175 cm. The duration between symptoms of brachial plexus paralysis and the diagnosis was 1-3 hours. All of these 7 patients presented various degrees of numbness in the ulnar side of the hand and forearm. Median nerve paresis was found in 3 cases and ulnar nerve paresis in 4 cases. Complete recovery of the neurological function had been achieved by the end of three months., Conclusions: The clinical features of brachial plexus palsy caused by halo traction include median nerve paresis, ulnar nerve paralysis, and numbness in the ulnar side of the hand and forearm, which may be due to the injury of the inferior part of the brachial plexus, i.e., damage of C8 and T1 nerve roots. Complete recovery of neurological function can be expected when the patient is kept under careful observation for recognizing this complication as soon as possible, then immediately reducing or removing the traction weight, and adopting rehabilitation training and neurotrophic pharmaceutical treatment.

Background: Non-viral methods of gene delivery, especially using polyethylenimine (PEI), have been widely used in gene therapy or DNA vaccination. However, the PEI system has its own drawbacks, which limits its applications., Methods: We have developed a novel non-viral delivery system based on PEI coated on the surface of bacterial magnetic nanoparticles (BMPs). The ability of BMPs-PEI complexes to bind DNA was determined by retardation of plasmid DNA in agarose gel electrophoresis. The transfection efficiency of BMPs-PEI/DNA complexes into eukaryotic cells was determined by flow cytometric analysis. The MTT assay was invited to investigate the cytotoxicity of BMPs-PEI/DNA complexes. The expression efficiency in vivo of BMPs-PEI bound to the plasmid pCMVbeta encoding beta-galactosidase was evaluated intramuscularly inoculated into mice. The immune responses of in vivo delivery of BMPs-PEI bound plasmid pcD-VP1 were determined by MTT assay for T cell proliferation and ELISA for detecting total IgG antibodies., Results: BMPs-PEI complexes could bind DNA and provide protection from DNase degradation. The transfection efficiency of BMPs-PEI/DNA complexes was higher than that in PEI/DNA complexes. Interestingly, in contrast to PEI, the BMPs-PEI complex was less cytotoxic to cells in vitro. We further demonstrated that the BMPs-PEI system can deliver an exogenous gene to animals and allow it to be expressed in vivo. Such expression resulted in higher levels of humoral and cellular immune responses against the target antigen compared to controls., Conclusions: We have developed a novel BMPs-PEI gene delivery system with a high transfection efficiency and low toxicity, which presents an attractive strategy for gene therapy and DNA vaccination., (Copyright 2007 John Wiley & Sons, Ltd.)

Despite their importance in activated processes, transition-event durations--which are much shorter than first passage times--have not received a complete theoretical treatment. The authors therefore study the distribution rhob(t) of durations of transition events over a barrier in a one-dimensional system undergoing overdamped Langevin dynamics. The authors show that rhob(t) is determined by a Fokker-Planck equation with absorbing boundary conditions and obtain a number of results, including (i) the analytic form of the asymptotic short-time transient behavior, which is universal and independent of the potential function; (ii) the first nonuniversal correction to the short-time behavior leading to an estimate of a key physical time scale; (iii) following previous work, a recursive formulation for calculating, exactly, all moments of rhob based solely on the potential function-along with approximations for the distribution based on a small number of moments; and (iv) a high-barrier approximation to the long-time (t-->infinity) behavior of rhob(t). The authors also find that the mean event duration does not depend simply on the barrier-top frequency (curvature) but is sensitive to details of the potential. All of the analytic results are confirmed by transition-path-sampling simulations implemented in a novel way. Finally, the authors discuss which aspects of the duration distribution are expected to be general for more complex systems.

Objective: To explore the effect of h(1) calponin mRNA expression on the biliary tract dynamics, and investigate the molecular mechanisms of gallstone formation in pregnancy., Methods: This study was carried out in Renmin Hospital of Wuhan University, and in the Department of Molecular Biology and Biochemistry, Wuhan University School of Medicine, Wuhan, China from July to December 2004. Thirty female guinea pigs were divided randomly into 3 groups, the nonpregnant group (n=10) (group A), the 30 days of pregnancy group (n=10) (group B), and the 60 days of pregnancy group (n=10) (group C). Animal models of pregnancy were established on pregnant group guinea pigs through feeding animals with one cage according to female versus male as 4:1. The total cholesterol (TC), total bilirubin (TBiL), total bile acid (TBA) in the bile and the serum estradiol (E(2)), progesterone (Pg) levels were determined respectively. Expression levels of h(1) calponin mRNA in gallbladder smooth muscles and Oddi's Sphincter (OS) were evaluated using semiquantitative reverse transcription polymerase chain reaction (RT-PCR)., Results: The concentration of TC, TBiL and the serum E(2) and Pg were more significantly increasing in group C than that in the other 2 groups. However, the concentration of TBA decreased gradually from group A to group C. Up-regulation of h(1) calponin gene expression was observed in the gallbladder smooth muscles in group C, but converse in OS., Conclusion: The h(1) calponin might play an important role in inducing dysfunction of extrahepatic biliary tract, bile stasis in gallbladder and gallstone formation in pregnancy.

Background: Compelling evidence indicates that Shigella species, the etiologic agents of bacillary dysentery, as well as enteroinvasive Escherichia coli, are derived from multiple origins of Escherichia coli and form a single pathovar. To further understand the genome diversity and virulence evolution of Shigella, comparative genomic hybridization microarray analysis was employed to compare the gene content of E. coli K-12 with those of 43 Shigella strains from all lineages., Results: For the 43 strains subjected to CGH microarray analyses, the common backbone of the Shigella genome was estimated to contain more than 1,900 open reading frames (ORFs), with a mean number of 726 undetectable ORFs. The mosaic distribution of absent regions indicated that insertions and/or deletions have led to the highly diversified genomes of pathogenic strains., Conclusion: These results support the hypothesis that by gain and loss of functions, Shigella species became successful human pathogens through convergent evolution from diverse genomic backgrounds. Moreover, we also found many specific differences between different lineages, providing a window into understanding bacterial speciation and taxonomic relationships.

In this study, we constructed single mutants MTS-1, MTS-2 of IroN and ShuA gene and double mutant MTS of them in Shigella dysenteriae A1 strain 51197 by insert and absence. The functional detection of every mutant was performed at the level of culture medium and cell experiment. The gene expression profiles of the mutants and the wild-type strains under iron-enriched and iron-limited conditions were analyzed by the SD51197 whole genomic microarray. The results showed that all the mutants grew obviously less well than the wild-type strains in L broth appending iron chelator DIP. The addition of iron to the cultures can stimulate the growth of mutants back to wild-type levels. In either the experiments on the ability of intracellular multiplication or the cell-to-cell spread in HeLa and U937 cell lines, mutants showed no obvious change in virulence compared with the parental strain SD51197. However when DIP was added to the cultured HeLa cells, the ability of intracellular multiplication of MTS-1, MTS-2, MTS has reduced about 23.4%, 25.2%, 43.6% respectively. The analysis of expression profiles under the iron-limited condition showed that the mutants were more sensitive for the changes of iron deficiency than the wild-type strains, many genes have been altered. Up-regulated genes mainly involved genes of transcription, coenzyme metabolism, amino acid transport and metabolism, and unknown functional genes, while down-regulated genes mainly involved genes of energy and carbohydrate metabolism and unknown function genes; the expression levels of known iron-transport associated genes generally showed up-regulated. The results demonstrated that iron-transport associated genes IroN, ShuA were likely to have some effects on the virulence and growth of S. dysenteriae.

Comparative Genomic Hybridization (CGH) microarray analysis was used to compare the genomic compositions of all eighteen Shigella boydii serotype representative strains. The results indicated the genomic "backbone" of this subgroup contained 2552 ORFs homologous to nonpathogenic E. coli K12. Compared with the genome of K12199 ORFs were found to be absent in all S. boydii serotype representatives, including mainly outer membrane protein genes and O-antigen biosynthesis genes. Yet the specific ORFs of S. boydii subgroup contained basically bacteriophage genes and the function unknown (FUN) genes. Some iron metabolism, transport and type II secretion system related genes were found in most representative strains. According to the CGH phylogenetic analysis, the eighteen S. boydii serotype representatives were divided into four groups, in which serotype C13 strain was remarkably distinguished from the other serotype strains. This grouping result corresponded to the distribution of some metabolism related genes. Furthermore, the analysis of genome backbone genes, specific genes, and the phylogenetic trees allowed us to discover the evolution laws of S. boydii and to find out important clues to pathogenesis research, vaccination and the therapeutic medicine development.

Genomic compositions of representatives of thirteen S. dysenteriae serotypes were investigated by performing comparative genomic hybridization (CGH) with microarray containing the whole genomic ORFs (open reading frames, ORFs) of E. coli K12 strain MG1655 and specific ORFs of S. dysenteriae A1 strain Sd51197. The CGH results indicated the genomes of the serotypes contain 2654 conserved ORFs originating from E. coli. However, 219 intrinsic genes of E. coli including those prophage genes, molecular chaperones, synthesis of specific O antigen and so on were absent. Moreover, some specific genes such as type II secretion system associated components, iron transport related genes and some others as well were acquired through horizontal transfer. According to phylogenic trees based on genetic composition, it was demonstrated that A1, A2, A8, A10 were distinct from the other S. dysenteriae serotypes. Our results in this report may provide new insights into the physiological process, pathogenicity and evolution of S. dysenteriae.

In asthma, human airway epithelial cells (HAECs) regulate the intensity of mucosal inflammation, in part, by releasing the pro-inflammatory cytokine interleukin (IL)-1beta. However, the IL-1beta inhibitors, IL-1 receptor antagonist (IL-1RA) and soluble IL-1 receptor type II (sIL-1RII), regulate IL-1beta bioactivity. In order to better understand the control of IL-1beta activity in the airway mucosa, the role(s) of tumour necrosis factor (TNF)-alpha, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in the release of IL-1beta and its inhibitors by cultured HAECs were examined. HAECs were treated with TNF-alpha (2-200 ng.mL(-1)), dibutyryl cAMP (0.01-1 mM), 8-bromo-cGMP (0.01-1 mM) or vehicle for 24 h, and cytokine levels in the HAEC-conditioned medium were measured by enzyme-linked immunosorbent assay. HAECs produced IL-1beta, IL-1RA and sIL-1RII constitutively, but the inhibitor concentrations greatly exceeded that of IL-1beta (by approximately 100- and approximately 550-fold, respectively). TNF-alpha dose-dependently increased the levels of all IL-1beta cytokine family members. However, over the range of TNF-alpha concentrations studied, IL-1beta concentration increased more than those of its inhibitors. cAMP increased constitutive and TNF-alpha-stimulated IL-1beta release but reduced that of sIL-1RII. In contrast, cGMP had no effect on IL-1beta but reduced IL-1RA and sIL-1RII release. Under basal conditions, the disproportionate release of inhibitors relative to interleukin-1beta by human airway epithelial cells probably prevents interleukin-1beta-mediated biological effects. Tumour necrosis factor-alpha, cyclic adenosine monophosphate and cyclic guanosine monophosphate may potentiate mucosal inflammation by increasing interleukin-1beta levels relative to those of its inhibitors in the airway mucosa.

A polysaccharide from the water extract of cultured Cordyceps militaris was isolated through ethanol precipitation, deproteination and gel-filtration chromatography. Their molecular weight was determined using gel-filtration chromatography. The structure of polysaccharide CPS-1 was elucidated by sugar analysis, Smith degradation, IR and 13C-NMR spectroscopy. CPS-1 was shown to possess a significant antiinflammatory activity and suppressed the humoral immunity in mice but had no significant effects on the cellular immunity and the non-specific immunity.

An experimental study of cellobiose inhibition in cellulose hydrolysis by synergism of cellobiohydrolyse I and endoglucanase I is presented. Cellobiose is the structural unit of cellulose molecules and also the main product in enzymatic hydrolysis of cellulose. It has been identified that cellobiose can strongly inhibit hydrolysis reaction of cellulase, whereas it has no effect on the adsorption of cellulase on cellulose surface. The experimental data of FT-IR spectra, fluorescence spectrum and circular dichroism suggested that cellobiose can be combined with tryptophan residue located near the active site of cellobiohydrolase and then form steric hindrance, which prevents cellulose molecule chains from diffusing into active site of cellulase. In addition, the molecular conformation of cellobiohydrolase changes after cellobiose binding, which also causes most of the non-productive adsorption. Under these conditions, microfibrils cannot be separated from cellulose chains, thus further hydrolysis of cellulose can hardly proceed.

The attenuated SA14-14-2 strain of Japanese encephalitis virus (JEV) was cultured in BHK-21 cells. The viral supernatant was purified and concentrated with PEG (MW 20,000). A suitable concentration of JEV antigen was used to sensitize latex to prepare the latex antigen. The specificity, sensitivity and stability of the antigen were assessed. A latex agglutination test (LAT) was developed for rapidly detecting antibody against JEV infection. The LAT and haemagglutination inhibition (HI) assay were compared by simultaneously testing 35 porcine serum samples from five farms. Ninety per cent (20/23) of the samples were seropositive by both assays. No significant difference was found between the two methods (p > 0.05). Furthermore, when 1,613 porcine sera from 120 farms were tested by LAT, the number of positive sera was 652, while that of negative sera was 961, ranging from 20% to 50% positive throughout the year. These results indicate that LAT is an appropriate candidate method for epidemiological surveys for and diagnosis of Japanese encephalitis.

Background: Activation of the beta(2)-adrenergic receptor (beta(2)AR) system expressed by human airway epithelial cells elicits a variety of cyclic adenosine monophosphate (cAMP)-dependent processes that help determine airway caliber and the intensity of airway inflammation in asthma. Glucocorticoids, mainstays in the treatment of asthma, profoundly affect the expression and function of the beta(2)-adrenergic receptor-adenylyl cyclase (beta(2)AR-AC) system in a variety of cell types. However, the effects of glucocorticoids on the beta(2)AR-AC system expressed by human airway epithelial cells are unstudied., Objective: We examined the effects of dexamethasone (DEX) on beta(2)AR gene expression and the function of the beta(2)AR-AC system in cultured human airway epithelial cells., Methods: Studies were performed in normal airway epithelial cells and BEAS-2B cells. Beta(2)AR gene expression was assessed from measurements of beta-adrenergic receptor density, beta(2)AR mRNA, and the activity of a full-length beta(2)AR promoter-luciferase reporter construct. The function of the beta(2)AR-AC system was assessed from cAMP production in response to the beta(2)-agonist isoproterenol and the expression of the stimulatory G protein G(alpha)s., Results: DEX had no effect on beta-adrenergic receptor density or on the beta(1)/beta(2) ratio over a wide range of concentrations and exposure times. However, DEX significantly but transiently enhanced beta(2)AR mRNA levels (approximately 1.5-fold) and beta(2)AR promoter activity (approximately 1.5-fold), indicating increased beta(2)AR gene transcription. DEX also dose-dependently enhanced cAMP responses to isoproterenol but not to forskolin, a direct activator of adenylyl cyclase. DEX-induced changes in cAMP production were associated with small (approximately 15%) increases in G(alpha)s expression., Conclusions: These data indicate that glucocorticoids only transiently enhance beta(2)AR gene transcription and fail to increase steady-state levels of beta(2)AR protein in human airway epithelial cells. Nonetheless, glucocorticoid-induced effects on the beta(2)AR-AC system of human airway epithelial cells contribute to the beneficial effects of corticosteroids in asthma by enhancing the functional response to beta(2)-agonists.

Interleukin (IL)-1 beta increases beta(2)-adrenergic receptor (beta(2)-AR) mRNA and density by protein kinase C (PKC)-dependent mechanisms in human airway epithelial cells. The present study examined the role of several nuclear transcription factors in the PKC-activated upregulation of beta(2)-AR expression. BEAS-2B cells were exposed to the PKC activator phorbol 12-myristate 13-acetate (PMA; 0.1 microM for 2-18 h). PMA had no effect on activator protein (AP)-2 or cAMP response element binding protein DNA binding activity but markedly increased nuclear factor (NF)-kappa B and AP-1 binding as assessed by electrophoretic gel mobility shift assay. PMA also increased the activity of a beta(2)-AR promoter-luciferase reporter construct in transiently transfected cells. These effects were inhibited by the PKC inhibitors Ro-31-8220 and calphostin C. Furthermore, with increasing Ro-31-8220, beta(2)-AR promoter-reporter activity correlated closely with both NF-kappa B and AP-1 activities (r > 0.89 for both). Finally, the selective NF-kappa B inhibitor MG-132 dose dependently reduced NF-kappa B binding and beta(2)-AR promoter activity but increased AP-1 binding. We conclude that PKC-induced upregulation of beta(2)-AR expression in human airway epithelial cells appears to be mediated, at least in part, by increases in NF-kappa B activity.

A conditional geometric phase shift gate, which is fault tolerant to certain types of errors due to its geometric nature, was realized recently via nuclear magnetic resonance (NMR) under adiabatic conditions. However, in quantum computation, everything must be completed within the decoherence time. The adiabatic condition makes any fast conditional Berry phase (cyclic adiabatic geometric phase) shift gate impossible. Here we show that by using a newly designed sequence of simple operations with an additional vertical magnetic field, the conditional geometric phase shift gate can be run nonadiabatically. Therefore geometric quantum computation can be done at the same rate as usual quantum computation.

Protein kinase C (PKC)-activated signal transduction pathways regulate cell growth and differentiation in many cell types. We have observed that interleukin (IL)-1beta upregulates beta2-adrenergic receptor (beta2-AR) density and beta2-AR mRNA in human airway epithelial cells (e.g., BEAS-2B). We therefore tested the hypothesis that PKC-activated pathways mediate IL-1beta-induced beta-AR upregulation. The role of PKC was assessed from the effects of 1) the PKC activator phorbol 12-myristate 13-acetate (PMA) on beta-AR density, 2) selective PKC inhibitors (calphostin C and Ro-31-8220) on beta-AR density, and 3) IL-1beta treatment on the cellular distribution of PKC isozymes. Recombinant human IL-1beta (0.2 nM for 18 h) increased beta-AR density to 213% of control values (P < 0.001). PMA (1 microM for 18 h) increased beta-AR density to 225% of control values (P < 0.005), whereas Ro-31-8220 and calphostin C inhibited the IL-1beta-induced upregulation of beta-AR in dose-dependent fashion. PKC isozymes detected by Western blotting included alpha, betaII, epsilon, mu, zeta, and lambda/iota. IL-1beta increased PKC-mu immunoreactivity in the membrane fraction and had no effect on the distribution of the other PKC isozymes identified. These data indicate that IL-1beta-induced beta-AR upregulation is mimicked by PKC activators and blocked by PKC inhibitors and appears to involve selective activation of the PKC-mu isozyme. We conclude that signal transduction pathways activated by PKC-mu upregulate beta2-AR expression in human airway epithelial cells.

In this study, the phylogenetic relationships between the two biovars and 14 serovars of Ureaplasma urealyticum were studied using the sequences of four different genes or genetic regions, namely: 16S rRNA genes; 16S-23S rRNA gene spacer regions; urease gene subunits ureA, ureB, partial ureC and adjoining regions upstream of ureA, ureA-ureB spacer and ureB-ureC spacer; the 5'-ends of the multiple-banded antigen (MBA) genes. U. urealyticum genotypes, based on all four genomic sequences, could be clearly separated into two clusters corresponding with currently recognized biovars 1 and 2. Sequences were generally conserved within each biovar. However, there was heterogeneity within the 5'-end regions of the MBA genes of the four serovars of biovar 1; the sequence of serovar 3 was identical with the previously published sequence and differed by only three bases from that of serovar 14; but there were significant differences between the sequences of serovars 3 and 14 and those of serovars 1 and 6. Based on the phylogenetic analysis, support is given to previous recommendations that the two biovars of U. urealyticum be classified as distinct species, namely U. parvum and U. urealyticum for biovars 1 and 2, respectively. In the future, the relationship between the new species and clinical manifestations of ureaplasma infections should be studied.

Twenty-seven cases of small bowel tumour confirmed surgically and pathologically have been studied. The sonographic appearances are described as well as a method of ultrasound detection following water ingestion. Tumours were classified into: (i) intraluminal tumours (duodenal adenocarcinoma n = 5; distal ileal lipoma with intussusception, n = 1; (ii) intramural tumours (proximal jejunal adenocarcinoma, n = 2; jejuno-ileal lymphoma, n = 5; (iii) extraluminal tumours (duodenal leiomyosarcoma, n = 2; neurilemmoma, n = 1; jejuno-ileal leiomyoma, n = 4; leiomyosarcoma, n = 6; subserosal lymphangioma, n = 1). Observing the lesion intermittently over a period of 1 h after water ingestion improves the localization of tumours within the small bowel.

Esculentoside A (EsA) is a saponin isolated from the roots of Phytolacca esculenta. Previous experiments showed that it had strong anti-inflammatory effects. Tumour necrosis factor (TNF) is an important inflammatory mediator. In order to study the mechanism of the anti-inflammatory effect of EsA, it was determined whether TNF production from macrophages was altered by EsA under lipopolysaccharide (LPS) stimulated conditions. EsA was found to decrease both extracellular and cell associated TNF production in a dose dependent manner at concentrations higher than 1 mumol/l EsA. Previous studies have showed that EsA reduced the releasing of platelet activating factor (PAF) from rat macrophages. The reducing effects of EsA on the release of TNF and PAF may explain its anti-inflammatory effect.