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The synthesis of the originally proposed structure of (±)-nicotlactone A, a potent antiviral lignan with three continuous chiral centers, is reported in 5 steps from methyl acrylate. The key steps of the synthesis included an In-catalyzed regioselective allylation and a Mn-catalyzed Mukaiyama hydration reaction. Our synthetic strategy also enabled us to get the other three epimers and investigate the structure–activity relationship. The NMR data of the synthesized compounds do not match that of the isolated sample, indicating that the structure of nicotlactone A remains to be reassigned. All the synthetic target compounds were evaluated for their anti-tobacco mosaic virus (anti-TMV) activity. Bioassay results indicated that (±)-8-demethylnicotlactone A displayed similar anti-TMV activity to the commercial agent ningnanmycin, thus being a promising candidate or lead compound for developing novel antiviral agents in crop protection.

Polynitrogen heterocycles are readily available and have recently arisen as versatile synthons for the formation of various C–C and C–X bonds, and medicinally active nitrogen-containing heterocycles. Several cascade reactions, including annulation, radical cascade, and borylation reactions, have been reported in which polynitrogen heterocycles are applied as arylation reagents. The success of these exceptional reactions illustrates the great synthetic potential of polynitrogen heterocycles, which provides a direct and useful approach to arylation reactions and the synthesis of nitrogen-containing heterocycles. The use of photocatalysts to effectively transfer energy from visible light to non-absorbing compounds has gained increasing attention as this method allows for the mild and efficient generation of radicals in a controlled manner. This approach has thus led to new methods that involve unique bond formation reactions. In addition, the use of free radical intermediates stabilized by transition metal catalysts is a powerful way to construct new chemical bonds. The aim of this review is to highlight the rapidly expanding area of radical-initiated denitrogenative cascade reactions of polynitrogen heterocycles and elaborate on their mechanisms from a new perspective by using photocatalysis and metal-based catalysis.

A novel and efficient strategy for trifluoromethylthiolation and dearomatization of activated alkynes with stable and readily available AgSCF3 has been developed. Reported herein is the unprecedented electrochemical generation of the SCF3 radical in the absence of persulfate for the synthesis of SCF3-containing spiro[5,5]trienones in good yields via a 6-exo-trig radical cyclization.

On the basis of the structure of nicotlactone A (

We previously demonstrated that circulating extracellular vesicles (EVs) from patients with valvular heart disease (VHD; vEVs) contain inflammatory components and inhibit endothelium-dependent vasodilation. Neutrophil chemotaxis plays a key role in renal dysfunction, and dexmedetomidine (DEX) can reduce renal dysfunction in cardiac surgery. However, the roles of vEVs in neutrophil chemotaxis and effects of DEX on vEVs are unknown. Here, we investigated the impact of vEVs on neutrophil chemotaxis in kidneys and the influence of DEX on vEVs. Circulating EVs were isolated from healthy subjects and patients with VHD. The effects of EVs on chemokine generation, forkhead box protein O3a (FOXO3a) pathway activation and neutrophil chemotaxis on cultured human umbilical vein endothelial cells (HUVECs) and kidneys in mice and the influence of DEX on EVs were detected. vEVs increased FOXO3a expression, decreased phosphorylation of Akt and FOXO3a, promoted FOXO3a nuclear translocation, and activated the FOXO3a signaling pathway in vitro. DEX pretreatment reduced vEV-induced CXCL4 and CCL5 expression and neutrophil chemotaxis in cultured HUVECs via the FOXO3a signaling pathway. vEVs were also found to suppress Akt phosphorylation and activate FOXO3a signaling to increase plasma levels of CXCL4 and CCL5 and neutrophil accumulation in kidney. The overall mechanism was inhibited in vivo with DEX pretreatment. Our data demonstrated that vEVs induced CXCL4-CCL5 to stimulate neutrophil infiltration in kidney, which can be inhibited by DEX via the FOXO3a signaling. Our findings reveal a unique mechanism involving vEVs in inducing neutrophils chemotaxis and may provide a novel basis for using DEX in reducing renal dysfunction in valvular heart surgery.

Acute heart failure (AHF) is a severe complication after cardiac surgery with cardiopulmonary bypass (CPB). Although some AHF biomarkers have been used in clinic, they have limitations when applied in the prediction and diagnosis of AHF after cardiac surgery with CPB, and there are still no effective and specific biomarkers. We and other researchers have shown that circulating microparticles (MPs) increased in a variety of cardiovascular diseases. However, whether the concentration of circulating MPs could be a new biomarker for AHF after cardiac surgery remains unknown. Here, 90 patients undergoing cardiac surgery with CPB and 45 healthy subjects were enrolled. Patients were assigned into AHF (n=14) or non-AHF (n=76) group according to the diagnosis criteria of AHF. The concentrations of circulating MPs were determined before, as well as 12 h and 3 days after operation with nanoparticle tracking analysis technique. MPs concentrations in patients before surgery were significantly higher than those of healthy subjects. Plasma levels of MPs were significantly elevated at 12 h after surgery in patients with AHF, but not in those without AHF, and the circulating MPs concentrations at 12 h after surgery were higher in AHF group compared with non-AHF group. Logistic regression analysis indicated that MPs concentration at postoperative 12 h was an independent risk factor for AHF. The area under receiver operating characteristic curve for MPs concentration at postoperative 12 h was 0.81 and the best cut-off value is 5.20×108 particles mL-1 with a sensitivity of 93% and a specificity of 10%. These data suggested that the concentration of circulating MPs might be a new biomarker for the occurrence of AHF after cardiac surgery with CPB.

A novel and efficient strategy for trifluoromethylthiolation and dearomatization of activated alkynes with stable and readily available AgSCF

The gene DTNBP1 encodes the protein dysbindin and is among the most promising and highly investigated schizophrenia-risk genes. Accumulating evidence suggests that dysbindin plays an important role in the regulation of neuroplasticity. Dysbindin was reported to be a stable component of BLOC-1 complex in the cytosol. However, little is known about the endogenous dysbindin-containing complex in the brain synaptosome. In this study, we investigated the associated proteome of dysbindin in the P2 synaptosome fraction of mouse brain. Our data suggest that dysbindin has three isoforms associating with different complexes in the P2 fraction of mouse brain. To facilitate immunopurification, BAC transgenic mice expressing a tagged dysbindin were generated, and 47 putative dysbindin-associated proteins, including all components of BLOC-1, were identified by mass spectrometry in the dysbindin-containing complex purified from P2. The interactions of several selected candidates, including WDR11, FAM91A1, snapin, muted, pallidin, and two proteasome subunits, PSMD9 and PSMA4, were verified by coimmunoprecipitation. The specific proteasomal activity is significantly reduced in the P2 fraction of the brains of the dysbindin-null mutant (sandy) mice. Our data suggest that dysbindin is functionally interrelated to the ubiquitin-proteasome system and offer a molecular repertoire for future study of dysbindin functional networks in brain.

The cysteine protease caspase-3, best known as an executioner of cell death in apoptosis, also plays a non-apoptotic role in N-methyl-d-aspartate receptor-dependent long-term depression of synaptic transmission (NMDAR-LTD) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor endocytosis in neurons. The mechanism by which caspase-3 regulates LTD and AMPA receptor endocytosis, however, remains unclear. Here, we addressed this question by using an enzymatic N-terminal peptide enrichment method and mass spectrometry to identify caspase-3 substrates in neurons. Of the many candidates revealed by this proteomic study, we have confirmed BASP1, Dbn1, and Gap43 as true caspase-3 substrates. Moreover, in hippocampal neurons, Gap43 mutants deficient in caspase-3 cleavage inhibit AMPA receptor endocytosis and LTD. We further demonstrated that Gap43, a protein well-known for its functions in axons, is also localized at postsynaptic sites. Our study has identified Gap43 as a key caspase-3 substrate involved in LTD and AMPA receptor endocytosis, uncovered a novel postsynaptic function for Gap43 and provided new insights into how long-term synaptic depression is induced.

Eight different strains of yeasts, Aspergillus niger , Aspergillus oryzae and White-rot fungus were tested for biotransformation of saponins to astragaloside IV in potato medium with Radix Astragali. Among these strains, immobilized cells of A. niger M85 exhibited the best performance on the biotransformation of saponins to astragaloside IV from Radix Astragali. After the biotransformation with immobilized A. niger M85, the contents of astragaloside IV and total astragalosides in Radix Astragali increased 10.7-fold (2.326 mg/g) and 8.6-fold (2.56 mg/g) to the solvent extraction sample, respectively. Optimization was done at incubation period 5 days, the amount of Radix Astragali powder 2 g, medium volume 50 mL/g, incubation temperature 30 °C and initial pH 6.0. The immobilized A. niger M85 can be reused seven times and retained 71% of its residual activity. The A. niger M85 was found to be a highly efficient producer of astragaloside IV.

The bacterial ribosomal protein S12 contains a universally conserved D88 residue on a loop region thought to be critically involved in translation due to its proximal location to the A site of the 30S subunit. While D88 mutants are lethal this residue has been found to be post-translationally modified to β-methylthioaspartic acid, a post-translational modification (PTM) identified in S12 orthologs from several phylogenetically distinct bacteria. In a previous report focused on characterizing this PTM, our results provided evidence that this conserved loop region might be involved in forming multiple proteins-protein interactions ( Strader , M. B. ; Costantino , N. ; Elkins , C. A. ; Chen , C. Y. ; Patel , I. ; Makusky , A. J. ; Choy , J. S. ; Court , D. L. ; Markey , S. P. ; Kowalak , J. A. A proteomic and transcriptomic approach reveals new insight into betamethylthiolation of Escherichia coli ribosomal protein S12. Mol. Cell. Proteomics 2011 , 10 , M110 005199 ). To follow-up on this study, the D88 containing loop was probed to identify candidate binders employing a two-step complementary affinity purification strategy. The first involved an endogenously expressed S12 protein containing a C-terminal tag for capturing S12 binding partners. The second strategy utilized a synthetic biotinylated peptide representing the D88 conserved loop region for capturing S12 loop interaction partners. Captured proteins from both approaches were detected by utilizing SDS-PAGE and one-dimensional liquid chromatography-tandem mass spectrometry. The results presented in this report revealed proteins that form direct interactions with the 30S subunit and elucidated which are likely to interact with S12. In addition, we provide evidence that two proteins involved in regulating ribosome and/or mRNA transcript levels under stress conditions, RNase R and Hfq, form direct interactions with the S12 conserved loop, suggesting that it is likely part of a protein binding interface.

In previous studies, we identified a putative 38-nucleotide stem-loop structure (zipcode) in the 3' untranslated region of the cytochrome c oxidase subunit IV (COXIV) mRNA that was necessary and sufficient for the axonal localization of the message in primary superior cervical ganglion (SCG) neurons. However, little is known about the proteins that interact with the COXIV-zipcode and regulate the axonal trafficking and local translation of the COXIV message. To identify proteins involved in the axonal transport of the COXIV mRNA, we used the biotinylated 38-nucleotide COXIV RNA zipcode as bait in the affinity purification of COXIV zipcode binding proteins. Gel-shift assays of the biotinylated COXIV zipcode indicated that the putative stem-loop structure functions as a nucleation site for the formation of ribonucleoprotein complexes. Mass spectrometric analysis of the COXIV zipcode ribonucleoprotein complex led to the identification of a large number RNA binding proteins, including fused in sarcoma/translated in liposarcoma (FUS/TLS), and Y-box protein 1 (YB-1). Validation experiments, using western analyses, confirmed the presence of the candidate proteins in the COXIV zipcode affinity purified complexes obtained from SCG axons. Immunohistochemical studies show that FUS, and YB-1 are present in SCG axons. Importantly, RNA immunoprecipitation studies show that FUS, and YB-1 interact with endogenous axonal COXIV transcripts. siRNA-mediated downregulation of the candidate proteins FUS and YB-1 expression in the cell-bodies diminishes the levels of COXIV mRNA in the axon, suggesting functional roles for these proteins in the axonal trafficking of COXIV mRNA.

In this study, two lentiviral vectors (LvNELL1 and LvBMP2) that encode human NELL1 and bone morphogenetic protein-2 (BMP2), respectively, were constructed. The effect of LvNELL1 infection on the proliferation, osteogenesis, and adipogenesis of human adipose-derived stem cells (hADSCs) in vitro was assessed and compared with that of LvBMP2. The results showed that hADSCs infected with LvNELL1 could efficiently and stably overexpress the target genes. 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide assay results demonstrated that LvBMP2, but not LvNELL1, enhanced the proliferation of hADSCs. Assessment of alkaline phosphatase activity and cellular mineralization indicated that LvNELL1 infection promoted the osteogenic differentiation of hADSCs, and the effect was comparable with that of LvBMP2. Real-time polymerase chain reaction (PCR) revealed that LvNELL1 infection upregulated OSX expression but not RUNX2 expression in hADSCs. In addition, adipogenic markers (lipid droplets, peroxisome proliferator-activating receptor g, and lipoprotein lipase) analysis showed that LvNELL1 could dramatically inhibit the adipogenic differentiation of hADSCs, but LvBMP2 had no such effect. Taken together, these findings suggested that lentiviral-mediated NELL1 gene transfer in hADSCs may be a novel and promising approach to achieve effective and precise bone regeneration.

NEL-like protein 1 (NELL1) is a newly identified secreted protein involved in craniosynostosis and has been found to promote osteogenic differentiation of mesenchymal stem cells. The objective of this study was to investigate the effect of NELL1 on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and the potential underlying mechanism. hPDLSCs underwent lentivirus-mediated NELL1 transfection (Lenti-NELL1) and markers of osteogenesis were assessed [alkaline phosphate (ALP), osteocalcin (OCN) and calcium deposition] to evaluate the effect of NELL1 on the differentiation of these cells. Quantitative polymerase chain reaction (qPCR) was employed to measure the mRNA expression of Msx2 and Runx2, and Lenti-enhanced green fluorescent protein (EGFP) served as a control. Western blot analysis and qPCR analyses confirmed that Lenti-NELL1-transfected hPDLSCs could express NELL1. Compared with the Lenti-EGFP group, ALP, OCN, calcium deposition and Msx2 mRNA expression were markedly increased (P0.01), but there was no significant difference in Runx2 mRNA expression between the two groups (P0.01). hPDLSCs can be transfected by Lenti-NELL1 and can stably express NELL1. NELL1 is able to promote the osteogenic differentiation of hPDLSCs, which may be related to the downregulation of Msx2 expression. Lenti-NELL1 transfection can be used during in vitro gene therapy for periodontal regeneration.

Radix Astragali is considered an excellent source of Traditional Chinese Medicine and a kind of famous restorative food. To make full usage of Radix Astragali, calycosin and formononetin were enriched through various macroporous resins from its residues, HPD500 resin, compared with other resins, offered the best effectiveness for the enrichment and separation of compounds. 35 °C was selected for its highest adoption capacity during static adsorption process, which can be obtained from adsorption isotherms. And then dynamic adsorption and desorption tests were carried out to optimize the operating parameters. After one run treatment with HPD500 resin, the contents of calycosin and formononetin in the product were 4.41-fold and 4.60-fold with recovery yields of 75.16% and 77.06%, respectively. Investigations on the antioxidant activity showed that eluate with 60% ethanol exhibited better free radical-scavenging activity (IC50 0.03 mg/mL) than crude extracts (IC50 0.05 mg/mL). Thus, this fast and effective method was established and could be a promising for the large-scale preparation of calycosin and formononetin from Radix Astragali residues.

International audience; Here, we examine tRNA-aminoacyl synthetase (ARS) localization in protein synthesis. Proteomics reveals that ten of the twenty cytosolic ARSs associate with ribosomes in sucrose gradients: phenylalanyl-RS (FRS), and the 9 ARSs that form the multi-ARS complex (MSC). Using the ribopuromycylation method (RPM) for localizing intracellular translation, we show that FRS and the MSC, and to a lesser extent other ARSs, localize to translating ribosomes, most strikingly when translation is restricted to poxvirus or alphavirus factories in infected cells. Immunoproximity fluorescence indicates close proximity between MSC and the ribosome. Stress induced-translational shutdown recruits the MSC to stress-granules, a depot for mRNA and translation components. MSC binding to mRNA provides a facile explanation for its delivery to translating ribosomes and stress granules. These findings, along with the abundance of the MSC (9 × 10(6) copies per cell, roughly equimolar with ribosomes), is consistent with the idea that MSC specificity, recently reported to vary with cellular stress (Netzer, N., Goodenbour, J. M., David, A., Dittmar, K. A., Jones, R. B., Schneider, J. R., Boone, D., Eves, E. M., Rosner, M. R., Gibbs, J. S., Embry, A., Dolan, B., Das, S., Hickman, H. D., Berglund, P., Bennink, J. R., Yewdell, J. W., and Pan, T. (2009) Nature 462, 522-526) can be modulated at the level of individual mRNAs to modify decoding of specific gene products.

The enrichment and separation of astragalosides I-IV (AGs I-IV) were studied on eight macroporous resins in the present study. SA-3 resin offered the best adsorption and desorption capacities for AGs I-IV than other resins. The models of adsorption kinetics were investigated in order to elucidate the mechanism of adsorption. The pseudo-second-order model was the better choice than the pseudo-first-order model to describe the adsorption behavior of AGs I-IV onto SA-3 resin. The equilibrium experimental data were well fitted to Langmuir and Freundlich isotherms. SA-3 resin adsorption chromatography tests were carried out to optimize the separation process of AGs I-IV from Radix Astragali extracts. With the optimum parameters for adsorption and desorption, the contents of AGs I-IV were 8.78-, 11.60-, 10.52- and 11.28-fold increased with the recovery yields being 65.88, 90.92, 84.25 and 94.17%, respectively. The preparative enrichment and separation of AGs I-IV from Radix Astragali extracts can be easily and effectively achieved by SA-3 resin adsorption chromatography. The developed methodology can also be referenced for the separation of other active constituents from herbal materials and manufacture of Radix Astragali products.

An efficient microwave-assisted extraction (MAE) technique was employed in extracting astragalosides I–IV from Radix Astragali. Astragalosides were quantified by liquid chromatography-electrospray ionisation mass spectrometry (LC-ESI/MS). The MAE procedure was optimised, validated and compared with other conventional extraction techniques. MAE gave the best result due to the highest extraction efficiency within the shortest extraction time. The optimal conditions of MAE were: employing 80% ethanol as solvent, ratio of solid/liquid 1:25 (g/ml), temperature 70 °C, irradiation power 700 W and three extraction cycles, each 5 min. Scanning electron microscopy (SEM) images of Radix Astragali materials after different extractions were obtained to provide visual evidence of the disruption effect. This is the first report on combining MAE with LC-ESI/MS for the extraction and quantification of astragalosides I–IV in Radix Astragali. The developed MAE method provided a good alternative for the extraction of triterpenoid saponins in Radix Astragali as well as other herbs.

MicroRNAs (miRNAs) selectively localize to subcompartments of the neuron, such as dendrites, axons, and presynaptic terminals, where they regulate the local protein synthesis of their putative target genes. In addition to mature miRNAs, precursor miRNAs (pre-miRNAs) have also been shown to localize to somatodendritic and axonal compartments. miRNA-338 (miR-338) regulates the local expression of several nuclear-encoded mitochondrial mRNAs within axons of sympathetic neurons. Previous work has shown that precursor miR-338 (pre-miR-338) introduced into the axon can locally be processed into mature miR-338, where it can regulate local ATP synthesis. However, the mechanisms underlying the localization of pre-miRNAs to the axonal compartment remain unknown. In this study, we investigated the axonal localization of pre-miR-338. Using proteomic and biochemical approaches, we provide evidence for the localization of pre-miR-338 to distal neuronal compartments and identify several constituents of the pre-miR-338 ribonucleoprotein complex. Furthermore, we found that pre-miR-338 is associated with the mitochondria in axons of superior cervical ganglion (SCG) neurons. The maintenance of mitochondrial function within axons requires the precise spatiotemporal synthesis of nuclear-encoded mRNAs, some of which are regulated by miR-338. Therefore, the association of pre-miR-338 with axonal mitochondria could serve as a reservoir of mature, biologically active miRNAs, which could coordinate the intra-axonal expression of multiple nuclear-encoded mitochondrial mRNAs.

Redundancy analysis (RDA) was employed to reveal the relationships between soil and rocky desertification through vegetation investigation and analysis of soil samples collected in typical karst mountain area of southwest Guizhou Province. The results showed that except TP, TK and ACa, all other variables including SOC, TN, MBC, ROC, DOC, available nutrients and basal respiration showed significant downward trends during the rocky desertification process. RDA results showed significant correlations between different types of desertification and soil variables, described as non-degradedpotential desertificationlight desertificationmoderate desertificationsevere desertification. Moreover, RDA showed that using SOC, TN, AN, and BD as soil indicators, 74.4% of the variance information on soil and rocky desertification could be explained. Furthermore, the results of correlation analysis showed that soil variables were significantly affected by surface vegetation. Considering the ecological function of the aboveground vegetation and the soil quality, Zanthoxylum would be a good choice for restoration of local vegetation in karst mountain area.

To investigate the influence of combined thoracoscopic and laparoscopic esophagectomy for early postoperative pulmonary function, and to study the relative factors for postoperative pulmonary complications.From September 2009 to December 2010, 61 patients with esophageal cancer had undergone esophagectomy surgery, of which 32 patients had undergone combined thoracoscopic and laparoscopic esophagectomy (CTLE group), and 29 patients had undergone open three-field esophagectomy (open group). Pulmonary function, including forced vital capacity (FVC), forced expiratory volume in 1 second (FEV(1)) were measured on the 1(th) preoperative day, 5(th) and 10(th) postoperative day, and arterial blood gas analyses were performed during the same period. Meanwhile, pain scores and other potentially relevant factors were recorded as well.Preoperative pulmonary function and arterial blood gas analysis, including FEV(1)%, FVC%, PaO2 in two groups had no significant difference (t = -1.608 to 0.709, P = 0.113 to 0.481). On the 10(th) postoperative day, FEV(1)%, FVC%, PaO2, and SaO2 of two groups were significantly different (FEV(1)%: 77% ± 17% vs. 53% ± 13%, t = 6.241, P = 0.000; FVC%: 78% ± 13% vs. 57% ± 16%, t = 5.549, P = 0.000; PaO2: (87 ± 9) mmHg vs. (79 ± 14) mmHg, t = 2.477, P = 0.017; SaO2: 96% ± 3% vs. 94% ± 2%, t = 2.313, P = 0.024; 1 mmHg = 0.133 kPa). Pain score of CTLE group was lower than open group, and the scores of two groups had significant difference before the 5(th) day after surgery (t = -4.398 to -1.815, P = 0.000 to 0.049). Postoperative pulmonary complications of CTLE group was lower than open group (6/32 vs. 12/29, χ(2) = 3.745, P = 0.049).Combined thoracoscopic and laparoscopic esophagectomy has advantages on early postoperative pulmonary function. It can relatively reduce the incidence of pulmonary complications after surgery.

β-methylthiolation is a novel post-translational modification mapping to a universally conserved Asp 88 of the bacterial ribosomal protein S12. This S12 specific modification has been identified on orthologs from multiple bacterial species. The origin and functional significance was investigated with both a proteomic strategy to identify candidate S12 interactors and expression microarrays to search for phenotypes that result from targeted gene knockouts of select candidates. Utilizing an endogenous recombinant E. coli S12 protein with an affinity tag as bait, mass spectrometric analysis identified candidate S12 binding partners including RimO (previously shown to be required for this post-translational modification) and YcaO, a conserved protein of unknown function. Transcriptomic analysis of bacterial strains with deleted genes for RimO and YcaO identified an overlapping transcriptional phenotype suggesting that YcaO and RimO likely share a common function. As a follow up, quantitative mass spectrometry additionally indicated that both proteins dramatically impacted the modification status of S12. Collectively, these results indicate that the YcaO protein is involved in β-methylthiolation of S12 and its absence impairs the ability of RimO to modify S12. Additionally, the proteomic data from this study provides direct evidence that the E. coli specific β-methylthiolation likely occurs when S12 is assembled as part of a ribosomal subunit.

To establish a stable animal model of temporomandibular joint (TMJ) synovitis.Sixteen 6-week-old male SD rats were classified into four groups, control group, occlusal dimension increase group, masseter resection group, occlusal dimension increase group and masseter resection group. The rats in the occlusal dimension increase group were adhered composite resin to their maxillary molars in order to increase the occlusal vertical dimension when they were 9-week-old. The rats in the masseter resection group were cut off their bilateral masseter muscles when they were 6-week-old. In the occlusal dimension increase group and masseter resection group, rats' bilateral masseter muscles were resected and occlusal vertical dimension was increased. All rats were sacrificed at their 10 weeks old. TMJ samples were prepared for histology to evaluate the animal model.The control group showed non-inflammatory changes. The occlusal dimension increase group and the masseter resection group showed vascular dilation and synovial lining proliferation, but there were no statistically significant differences between the two groups (P0.05). Compared to the two disposed groups, the occlusal dimension increase group and masseter resection group showed significant inflammatory changes (P0.05), including synovial lining proliferation, vascular dilation and fibrin deposit.The animal model of TMJ synovitis created in the present investigation could simulate the real pathological features of synovitis in vivo, and this animal model showed the obvious merits of high stability and reproduction.

The optimal conditions for extraction of astragalosides III and IV (AGs III and IV) in Radix Astragali by negative pressure cavitation-accelerated enzyme pretreatment were studied on the basis of a Box-Behnken design and response surface methodology. Experimental results showed that negative pressure, amount of enzyme and incubation temperature were the main factors governing the enzyme pretreatment of Radix Astragali. The optimum parameters were obtained as follows: negative pressure -0.08 Mpa, amount of enzyme 1.48% (w/w of materials) and incubation temperature 45 degrees C. Under the optimal conditions, the maximal extraction yields of AGs III and IV were 0.103 and 0.325 mg/g, which were 41.67% and 65.31% increased as compared to those without enzyme pretreatment, respectively. The effect of negative pressure cavitation and enzyme pretreatment on the structural changes of plant cells was observed by scanning electron microscopy. In conclusion, negative pressure cavitation-accelerated enzyme pretreatment was proved to be environment-friendly and economical, and could be used in secondary metabolites production.

Discretization of attributes with real values in rough set is an important problem in data mining. It is different from the traditional discretization which has particular characteristic. Nguyen S. H has given a detailed description about discretization in rough set. This paper gives a genetic algorithm aimed at discretization proposed by Nguyen S. H. And at the same time we make an experiment on several datasets from UCI machine learning repository by this method. During the experiment, we constantly optimized genetic algorithm by adopting some optimization strategies. The experiment has proved that using genetic algorithm to solve discretization of rough set is efficient whatever in time complexity or in accuracy.

The concept (or Galois) lattice produced from a binary relation which has been proved is useful for many applications. But building concept lattice is difficult. Reference [L. Nourine, et al., 1999] gives a fast algorithm for building lattice, but the concept lattice has some special characters. This article presents a fast algorithm for building concept lattice and corresponding graph, an also gives the time complexity of this algorithm. Database updating is always necessary. In our paper, we also give an algorithm for modify lattice when data update happened.

To demonstrate that mitochondrial morphological and functional changes are an important intermediate link in the course of apoptosis in esophageal carcinoma cells induced by As2O3.The esophageal carcinoma cell line SHEEC1, established in our laboratory, was cultured in 199 growth medium, supplemented with 100mL x L(-1) calf serum and 3 mol x L(-1)As2O3 (the same below). After 2, 4, 6, 12, 24 h of drug adding, the SHEEC1 cells were collected for light-and electron-microscopic examination. The mitochondria were labeled by Rhodamine fluorescence probe and the fluorescence intensity of the mitochondria was measured by flow cytometer and cytofluorimetric analysis. Further,the mitochondrial transmembrane potential (MTP, psim) change was also calculated.The mitochondrial morphological change after adding As2O3 could be divided into three stages. In the early-stage (2-6h) after adding As2O3, an adaptive proliferation of mitochondria appeared; in the mid-stage (6-12 h) a degenerative change was observed; and in the late-stage (12-24 h) the mitochondria swelled with outer membrane broken down and then cells death with apoptotic changes of nucleus. The functional change of the mitochondria indicated by fluorescent intensity, which reflected the MTP status of mitochondria, was in accordance with morphological change of the mitochondria. The fluorescent intensity increased at early-stage, declined in mid-stage and decreased to the lowest in the late-stage. 24 h after As2O3 adding, the cell nucleus showed typical apoptotic changes.Under the inducement of As2O3, the early apoptotic changes of SHEEC1 cells were the apparent morphological and functional changes of mitochondria, afterwards the nucleus changes followed. It is considered that changes of mitochondria are an important intermediate link in the course of apoptosis of esophageal carcinoma cells induced by As2O3.