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Omnivorous insects make foraging decisions between plant and prey resources depending on their accessibility, availability, and nutritional value. This shapes the stability and complexity of foodwebs, but also pest control services in agroecosystems. The mirid bug Lygus pratensis is a common cotton pest in China, but it also feeds on a variety of prey species. However, little is known about how different types of available resources affect its fitness and foraging behaviour. In laboratory experiments, we measured the fitness (survival, longevity and fecundity) of L. pratensis fed with bean pod only, bean + Aphis gossypii nymphs, or bean + Helicoverpa armigera eggs, and we also conducted focal observations of its foraging behaviour when provided the latter two. Adding H. armigera to its diet increased its fitness (both survival and fecundity), while adding A. gossypii was marginally detrimental. The different diets did not affect the time spent walking (searching for food resources) or preying, but significantly affected the time spent sapping bean tissue depending on L. pratensis life stage. Nymphs spent more time sapping plant when provided with H. armigera than A. gossypii (possibly through higher efficiency of handling prey). In addition, adults spent less time sapping plant than did nymphs (possibly through good efficiency of handling A. gossypii mobile prey and due to higher protein requirements). This special case of life-history omnivory highlights the complexity of natural foodwebs, where a major pest at the juvenile stage may reduce pest damage by preying on other pests species at the adult stage.

This study aimed to compare the efficiencies of clustered regulatory interspaced short palindromic repeat (CRISPR)/Cas9-mediated gene knock-ins with zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) in bovine and dairy goat fetal fibroblasts. To test the knock-in efficiency, a set of ZFNs and CRISPR/Cas9 plasmids were designed to edit the bovine myostatin (MSTN) gene at exon 2, while a set of TALENs and CRISPR/Cas9 plasmids were designed for editing the dairy goat β-casein gene at exon 2. Donor plasmids utilizing the ZFNs, TALENs, and CRISPR/Cas9 cutting sites were constructed in the GFP-PGK-NeoR plasmid background, including a 5’ and 3’ homologous arm flanking the genes humanized Fat-1 (hFat-1) or enhanced green fluorescent protein (eGFP). Subsequently, the ZFNs, TALENs, or CRISPR/Cas9 and the hFat-1 or eGFP plasmids were co-transfected by electroporation into bovine and dairy goat fetal fibroblasts. After G418 (Geneticin) selection, single cells were obtained by mouth pipetting, flow cytometry or a cell shove. The gene knock-in events were screened by PCR across the homologous arms. The results showed that in bovine fetal fibrobalsts, the efficiencies of ZFNs-mediated eGFP and hFat-1 gene knock-ins were 13.68 and 0%, respectively. The efficiencies of CRISPR/Cas9-mediated eGFP and hFat-1 gene knock-ins were 77.02 and 79.01%, respectively. The eGFP gene knock-in efficiency using CRISPR/Cas9 was about 5.6 times higher than when using the ZFNs gene editing system. Additionally, the hFat-1 gene knock-in was only obtained when using the CRISPR/Cas9 system. The difference of knock-in efficiencies between the ZFNs and CRISPR/Cas9 systems were extremely significant (P

Nowadays, strain online monitoring technology of high-temperature components plays an important role in ensuring accurate operation and safe production in the fields of aerospace, nuclear power and power plants. A SAW high-temperature strain sensor employing composite structure of based Al 2 O 3 / IDTs/ AlN/ Metal/ Al 2 O 3 is proposed to obtain strain information at extreme high-temperature environment. Obviously, high performance as Q-value and low insertion loss of the sensing chip is the key for achieving successful wireless strain sensing system. In this work, the theoretical simulation by using the finite element method (FEM) analysis was performed to realize the SAW propagation in multi-layered composite structures, and corresponding coupling of modes (COM) parameter was extracted. On this basis, the optimal design on the SAW high-temperature strain sensing device was realized.

In this study, a central composite rotatable design based on response surface methodology (RSM) was employed to design and formulate an appropriate paeonol microparticle formulation. Five levels of a three-factor, rotatable, central composite design were used to evaluate the critical formulation variables. The optimum conditions for preparing paeonol-loaded microparticles were predicted to be: polyvinyl alcohol (PVA) content (2.84%), the ratio of drug to polymer (6.88) and the stirring rate (1007.59 rpm). The optimized responses for production yield and loading efficiency were found to be 68.86% and 55.90%, respectively, and the particle size were 23.27 ± 0.76 µm and the sorting coefficient (σ) was 0.732. Furthermore, in vitro release study suggested that microparticle could be a suitable delivery system in treating skin disease for its sustained release of drug. In conclusion, RSM can be successfully used to optimize the effect of formulation variables.

Background Percutaneous coronary interventions to implant bioresorbable vascular scaffolds (BVSs) were designed to reduce the late thrombotic events that occur with metallic stents. Purpose To estimate the incidence of scaffold thrombosis after BVS implantation and compare everolimus-eluting BVSs with everolimus-eluting metallic stents (EESs) in terms of safety and efficacy at mid- and long-term follow-up in adults who had a percutaneous coronary intervention. Data sources PubMed, EMBASE, the Cochrane Library, conference proceedings, and relevant Web sites from inception until 20 May 2017, without language restriction. Study selection 7 randomized trials and 38 observational studies (each with a minimum of 6 months and 100 patient-years of follow-up) in adults with coronary artery disease who had a BVS or an EES and reported scaffold or stent thrombosis (main outcome) or other secondary outcomes (such as death, myocardial infarction, or revascularization). Data extraction 2 reviewers independently extracted study data, rated study quality, and assessed strength of evidence. Data synthesis The pooled incidence of definite or probable scaffold thrombosis after BVS implantation was 1.8% (95% CI, 1.5% to 2.2%) at a median follow-up of 1 year (41 studies, 21 884 patients) and 0.8% (CI, 0.5% to 1.3%) beyond 1 year (14 studies, 4688 patients). Seven trials involving 5578 patients that directly compared BVSs with EESs showed an increased risk for definite or probable scaffold thrombosis (odds ratio [OR], 3.40 [CI, 2.01 to 5.76]) with BVSs at a median follow-up of 25 months. Increased risks were present at early (prominently subacute), late, and very late stages, and odds beyond 1 year were almost double those seen within 1 year. Bioresorbably vascular scaffolds increased risks for myocardial infarction (OR, 1.63 [CI, 1.26 to 2.10]), target lesion revascularization (OR, 1.31 [CI, 1.03 to 1.67]), and target lesion failure (OR, 1.37 [CI, 1.12 to 1.66]); the odds for these 3 end points also increased over time. The incidences of all-cause, cardiac, and noncardiac death and of target vessel and any revascularization did not differ. Limitation Quality of observational studies was unclear, and some data were unpublished. Conclusion Compared with EESs, BVSs increased the risks for scaffold thrombosis and other thrombotic events at mid- and long-term follow-up, and risks increased over time. Primary funding source National Natural Science Foundation of China.

Atomic force microscopy (AFM) is a promising microscopy technique that can provide high-resolution images of bacterial cells without fixation. Three species of bacteria, Xanthomonas campestris, Pseudomonas syringae, and Bacillus subtilis, were used in this study. AFM images were obtained from unfixed and glutaraldehyde-fixed cells, and cell height was measured. The mean height of bacterial cells prepared by fixation was higher than that of those prepared by nonfixation. However, the height changes were different between Gram-negative and Gram-positive bacteria: the mean height of two fixed Gram-negative bacteria, X. campestris and P. syringae, increased by 112.31 and 84.08%, respectively, whereas Gram-positive bacterium, B. subtilis, increased only by 38.79%. The results above indicated that glutaraldehyde fixation could affect the measured height of cells imaged by AFM; further more, the effect of glutaraldehyde fixation on the measured height of Gram-negative bacterial cells imaged by AFM seemed much more than on that of Gram-positive bacterial cells. SCANNING 34: 6–11, 2012. © 2011 Wiley Periodicals, Inc.

The purpose of this investigation was to compare the effects of different relative humidity (RH) on the microcosmic conformation of the recombinant AAV-2 virion at 22°C. rAAV-2 virions prepared on copper grid were placed in a high, middle or low RH cabinet and incubated for 72, 48 and 24 h, respectively. The rAAV-2 virions were observed by transmission electron microscope and the values of major axis length, minor axis length and ellipticity of the rAAV-2 virions were obtained using the IMS cell image analysis system. After incubation for 48 and 72 h, the major axis length and minor axis length of the rAAV-2 virion started to rapidly decrease in high RH. Conversely, the axis lengths rapidly increased in low RH. Then, the ellipticity of the rAAV-2 virion would almost tend to approach the identical value of 0.9 for 48 and 72 h incubations in high RH. The results suggest that the rAAV-2 virion tended to favor a smaller, round, more stable conformation in high RH compared to low RH which implied that the rAAV-2 virion was probably prone to living in high relative humidity conditions.

Insulin-like growth factor type-I receptor (IGF-1R) signal pathway is critical in several physiological activities and some pathogenesis. IGF-1R derived nonphosphorylated peptide LYASVNPEY was immobilized on the surface of CM5 sensor chip specialized for surface plasmon resonance (SPR) biosensor. The peptide surface was treated with cell lysates containing or not containing IGF-1R kinase. The results show that using SPR biosensor PY20 can discriminate the different levels of peptide tyrosine phosphorylation (PTP) caused by different incubation times and different lysates. The effect of the PTP on peptide-protein interaction was further studied. The results demonstrated that only the phosphorylated peptide can interact with its downstream insulin receptor substrate-1 (IRS-1). The binding affinity (KA) and the association and dissociation kinetics (ka and kd) of the phosphorylated peptide with IRS-1 were measured by SPR biosensor, with the results of KA = (2.02 ± 0.08) × 108 M−1, ka = (2.30 ± 0.15) × 106 M−1 s−1 and kd = (1.14 ± 0.13) × 10−2 s−1, respectively. This new phosphorylation method of peptide that simulates cell receptor has the characteristics of real-time, speediness, label-free, and convenience for detection, which is instrumental for investigations on receptor phosphorylation signal pathways and drug screening against these signal pathways.

The paeonol proniosomes ointment and ordinary ointment were administered to rats. Physiological saline served as perfused solution. The perfusion rate was 5 mL x L(-1) and the microdialysis samples were collected every 20 min intervals. The paeonol concentration in perfused solution was determined by HPLC. Investigation of the pharmacokinetics of paeonol proniosomes ointment and ordinary ointment by the skin-blood synchronous microdialysis coupled with HPLC is reported in this study. The results show that the recovery was (54.80 +/- 1.50)% in vitro and (54.58 +/- 4.61)% in vivo. The results showed that paeonol proniosomes ointment significantly raised the drug concentrations in skin more than the paeonol ordinary ointment. The paeono proniosomes ointment has less drugs into the blood as the ordinary ointments in blood, but its blood drug concentrations were steadier. The paeonol proniosomes ointment may be developed into a new preparation.

In this paper we try to find out the importance of legitimacy across different stages of corporate life cycles. We introduce the concept of ties intensity and develop a conceptual framework showing that at the enterprise's different development stages, how the two different kinds of ties intensity will affect the legitimacy types. Our research is devoted to find out what kind of effect will different development stages have on legitimacy. We find that in enterprise creation period, the strength of market ties has positive effect on the legitimacy, but the strength of government ties has no significant effect on it; In the early growth period of enterprises, the strength of market ties has no significant effect on the legitimacy, but the strength of government ties has positive effect it; In the mature period of the enterprise, the strength of market ties has positive effect on the legitimacy, the strength of government ties has no significant effect on it.

To clone 1b type of HCV NS3-4b Gene and express in HEK 293 cells, lay the foundation for further study of the HCV NS3-4b recombinant adeno-associated virus vaccine and its dendritic cell vaccine.HCV 1b patients' serum was collected, and full length NS3-4b segment was amplified by RT-PCR and cloned into adeno-associated virus' expression vector pAAV. CMV. EGFP in order to express in HEK 293 cells. At last, it was validated whether express or not by Western Blot.The 1b type gene NS3-4b were amplified and consistent to the expected size (2838 bp), the recombinant plasmid has been confirmed its successful restructured by double enzyme and sequencing, at last, Western Blot map can see objective protein expression after it transfect HEK 293 cells.The adeno-associsted virus recombination HCV NS3-4b plasmid have successfully constructed and it can express in eukaryotic cells.

The aim of the present study was to design a novel topical skin-target drug-delivery system, the paeonol microsponge, and to investigate its drug-release patterns in dosage form, both in vitro and in vivo. Paeonol microsponges were prepared using the quasi-emulsion solvent-diffusion method. In vitro release studies were carried out using Franz diffusion cells, while in vivo studies were investigated by microdialysis after the paeonol microsponges were incorporated into a cream base. In vitro release studies showed that the drug delivered via microsponges increased the paeonol permeation rate. Ex vivo drug-deposition studies showed that the microsponge formulation improved drug residence in skin. In addition, in vivo microdialysis showed that the values for the area under the concentration versus time curve (AUC) for the paeonol microsponge cream was much higher than that of paeonol cream without microsponges. Maximum time (Tmax) was 220 min for paeonol microsponge cream and 480 min for paeonol cream, while the half-life (t1/2) of paeonol microsponge cream (935.1 min) was almost twice that of paeonol cream (548.6 min) in the skin (n = 3). Meanwhile, in the plasma, the AUC value for paeonol microsponge cream was half that of the paeonol cream. Based on these results, paeonol-loaded microsponge formulations could be a better alternative for treating skin disease, as the formulation increases drug bioavailability by lengthening the time of drug residence in the skin and should reduce side-effects because of the lower levels of paeonol moving into the circulation.

The envelope protein of dengue virus is involved in host cell attachment for entry and induction of protective immunity. Current efforts are focused on producing a tetravalent vaccine by mixing four monovalent vaccine components. In this work, we developed a genetic vaccine based on a novel adeno-associated viral (AAV) vector expressing the carboxy-terminal truncated envelope protein (79E) of dengue virus. The expression of the recombinant 79E protein in HEK 293 cells was confirmed by Western blot. Vectors packaged with novel AAV capsids (AAV2/8 or AAV2/rh32.33) were injected into C57BL/6 mice intramuscularly. Dengue virus antigen was produced in the mice and induced long-lasting antibody responses against the dengue virus still detectable 20 weeks after immunization. AAV2/8 vaccine induced higher anti-dengue virus antibody levels than AAV2/rh32.33 vaccine or AAV plasmid. Furthermore, the anti-dengue antibodies could neutralize homogeneous dengue virus. These results demonstrated that the AAV vaccines possessed appropriate immunogenicity and could be used for the development of an effective dengue vaccine.

To investigate the possible role of perforin (PFN) in the pathogenesis of severe preeclampsia.Thirty-two cases of severe preeclampsia were included in the study. Thirty-two cases of normal pregnancy were selected as control group in random. The expression of PFN mRNA in the peripheral blood mononuclear cells (PBMC) was detected by reverse transcription (RT)-PCR, and its correlation with mean arterial pressure was analyzed in severe preeclamptic patients. The expression of PFN protein in the decidua was detected by immunohistochemistry.(1) The expression of PFN mRNA in PBMC:the PFN mRNA level in severe preeclamptic group was 1.19 ± 0.31, and that in normal pregnancy group is 0.82 ± 0.28. The PFN mRNA level in severe preeclamptic group was significantly higher than that of control group (P0.01). (2) Correlation analysis: the mean blood pressure in severe preeclampsia group was (133 ± 5) mm Hg (1 mm Hg = 0.133 kPa). There was significant positive correlation between level of PFN mRNA in PBMC and mean blood pressure in severe preeclamptic patients (r = 0.701, P = 0.000). (3) Decidual PFN protein expression:PFN protein was mainly expressed in lymphocytes and the cytoplasm of decidual stromal cells. The positive ratio of PFN in the decidua of severe preeclamptic patients was 84% (27/32), significantly higher than that of control group (53%, 17/32, P0.01).Expression of PFN was significantly increased in severe preeclampsia, and it was of significant positive correlation with mean blood pressure. PFN may participate in the pathogenesis of severe preeclampsia.

Colletotrichum capsici f. nicotianae is an important plant pathogen in tobacco-grown area of Weifang region of Shangdong Province, China. In this study, the toxicity of liquid culture media from different isolates was characterized, and some properties of the toxic ingredient were identified. The results indicated that the optimal toxin-producing conditions for C. capsici f. nicotianae were in potato dextrose broth under pH 6.0, at 25∼30 °C for 13 days. The liquid culture media from all isolates were toxic to tobacco plants and induced the wilting symptoms. The toxin from the liquid culture media has thermal, acid–base stability and a broad spectrum of toxicity to the plants. Furthermore, the direct bioassay for two components of the liquid filtrates precipitated by ethanol showed that the active ingredient of the toxin is a kind of nonprotein substance, which was further supported by the papain hydrolysis test.

Hydroelectric cascade exploitation, one of the major ways for exploiting water resources and developing hydropower, not only satisfies the needs of various national economic sectors, but also promotes the socio-economic sustainable development of river basin. unavoidable anthropogenic impacts on the entire basin ecosystem. Based on the process of hydroelectric cascade exploitation and the ecological characteristics of river basins, this paper reviewed the major impacts of hydroelectric cascade exploitation on dam-area ecosystems, river reservoirs micro-climate, riparian ecosystems, river aquatic ecosystems, wetlands, and river landscapes. Some prospects for future research were offered, e.g., strengthening the research of chain reactions and cumulative effects of ecological factors affected by hydroelectric cascade exploitation, intensifying the study of positive and negative ecological effects under the dam networks and their joint operations, and improving the research of successional development and stability of basin ecosystems at different temporal and spatial scales.

Atomic force microscopy (AFM) is a promising microscopy technique that can provide high-resolution images of bacterial cells without fixation. Three species of bacteria, Xanthomonas campestris, Pseudomonas syringae, and Bacillus subtilis, were used in this study. AFM images were obtained from unfixed and glutaraldehyde-fixed cells, and cell height was measured. The mean height of bacterial cells prepared by fixation was higher than that of those prepared by nonfixation. However, the height changes were different between Gram-negative and Gram-positive bacteria: the mean height of two fixed Gram-negative bacteria, X. campestris and P. syringae, increased by 112.31 and 84.08%, respectively, whereas Gram-positive bacterium, B. subtilis, increased only by 38.79%. The results above indicated that glutaraldehyde fixation could affect the measured height of cells imaged by AFM; further more, the effect of glutaraldehyde fixation on the measured height of Gram-negative bacterial cells imaged by AFM seemed much more than on that of Gram-positive bacterial cells.

In this study, we propose a new method to predict hairpins in proteins and its evaluation based on the support vector machine. Different from previous methods, new feature representation scheme based on auto covariance is adopted. We also investigate two structure properties of proteins (protein secondary structure and residue conformation propensity), and examine their effects on prediction. Moreover, we employ an ensemble classifier approach based on the majority voting to improve prediction accuracy on hairpins. Experimental results on a dataset of 1926 protein chains show that our approach outperforms those previously published in the literature, which demonstrates the effectiveness of the proposed method.

To evaluate the effects of bruxism on masticatory muscle electromyographic (EMG) activity.Twenty-four bruxers and sixteen asymptomatic control subjects were included through questionnaire and clinical examination. EMG activity was recorded by placing surface electrodes on bilateral anterior temporalis (TA), masseters (MM), anterior digastrics (DA) and sternocleidomastoid (SCM) muscles. EMG activities at rest, during maximal voluntary clenching in intercuspal position and swallowing were recorded by means of Bio PAK system.EMG activities of TA and MM at rest were significantly higher in bruxism group than in control group (P0.05). When subjects clenched their teeth in intercuspal position, the activities of TA and MM were much lower in bruxism group than in control one (P0.05). EMG activity during swallowing was no significant difference between the two groups. The asymmetry index of bilateral TA and MM in bruxism group was a little higher than the control group, but there was no significant difference between the two groups (P0.05).Masticatory muscle dysfunction of bruxers is mainly represented as higher potential in postural position and lower potential during maximal voluntary clenching in intercuspal position of anterior temporalis and masseters.

Rat fetal neural stem cells (rFNSCs) was separated from embryo about 14.5-16.5 days, and cultured in DMEM/F12 media with additives and epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). Effects of lipid on growth and proliferation of rFNSCs was examined by counting the number of neurospheres and incorporation of 3H. The data show that chemical defined lipid improved rFNSCs' growth and cell division. Lipid will be another neural stem cell's culture media's additive.

To investigate the effects of palmitic acid (PA) on human hepatocytes and its mechanism.We administered a mimic hyperlipidemia condition of 0.2-0.4 mmol/L PA to human hepatoma cell line, HepG2 cells. Cell viability was determined by Trypan blue staining. Cell cycle and early apoptosis were determined by propidium iodide and/or Annexin V staining, and the levels of Bcl-2 and Bax were analyzed by flow cytometry.An inhibition of cell growth was observed at a dose- and time-dependent manner in HepG2 cells after the treatment of PA. An apoptosis with appearance of sub-G1 fraction determined by cell cycle analysis significantly increased after the treatment of PA for 4 days. Bcl-2 level slightly decreased; in contrast, Bax level elevated markedly, which resulted in a significant decrease of Bcl-2/Bax ratio.PA may induce cell death on hepatocytes via mitochondria-mediated apoptosis by reducing the level of Bcl-2/Bax.

The 3.0 kb 5' arm (long arm, LA) of rat HPRT gene knock-out vector was cut by Sal I from rat HPRT gene genomic bacterial artificial chromosome (BAC) , and the 1.7 kb 3' arm (short arm, SA) was proliferated by PCR. Neo', 5' arm, 3' arm were sequentially cloned into pBS vector's relative restriction enzyme sites. For acquirement of tk gene, the 5' arm -Neo' -3' arm fragment inserted into pKO vector to construct pKO-HPRT. The pKO-HPRT was linearized by Not I, extracted by ethidium bromide, butanol and phenol/chloroform, and dialyzed by 0.025 microlmol/L Millipore. At the same time, rat neural stem cells cultured from E14.5-16.5 rat fetal brain. Passage 2 rFNSCs was tranfected by linearized pKO-HPRT with Fugene-6t transfection reagents. After 80 microg/ml G418 and 0.2 micromol/L ganciclovir selection, the survived cells was cultured in suspension to form neural spheres. The spheres can be picked up under the microscopy, and proliferated in 96- ,48- and 24-well plates sequentially. When the cell number reached 4 x 10(3)/well, half cells was lysed by lysis buffer to extract DNA, the other half was kept on growing to freeze and extract RNA. The knock-out cell colonies first detected by PCR, then confirmed by Southern blot and RT-PCR. All the results show that we have knocked out HPRT gene in three rat fetal neural stem cell colonies from 32 colonies.

To prepare monoclonal antibodies (mAb) against recombinant human nucleoside diphosphate kinase-A(NDPK-A) and characterize their properties.BALB/c mice were immunized with rhNDPK-A, and mAb was prepared by hybridoma technique. Ig subclass and specificity was analysed by double immunodiffusion and western blot respectively. The titres of mAbs in ascitic fluid, relative affinity and epitopes recognized by mAbs were determined by indirect ELISA.6 hybridoma cell lines secreting anti-rhNDPK-A mAbs were obtained. Their Ig subclass belonged to IgG1. The titers of 6 mAbs in ascitic fluid were 1x10(-4)-5x10(-6). Relative affinity of mAbs were 4.5x10(-9)-2.8x10(-10) mol/L. They recognized 3 different epitopes on rhNDPK-A molecule.6 mAbs against rhNDPK-A have been prepared successfully which provide useful reagent for clinical diagnosis and further research.

SOX2, a SRY-related HMG box protein, is thought to be an important transcription factor during organogenesis, including the stomach although the expression and function are unclear. We investigated SOX2 protein expression to clarify its roles in differentiation and carcinogenesis of the stomach. Using polyclonal SOX2 antibodies, expression of SOX2 in gastric normal mucosae, intestinal metaplasia and carcinomas from 68 gastric carcinoma patients was studied by immuohistochemistry. SOX2 was strongly and moderately expressed in the nuclei of the foveolar epithelium and intestinal metaplasia, respectively, the expression being much higher than that in carcinomas (p