A study on the effect of the bacterial inoculant on corn silage quality,digestibility and performance in dairy cattle

Effect of the bacterial inoculant Sil-All (short for ‘ SA‘ ) on corn silage quality, digestibility and performance in dairy cattle was studied. SA treated silage resulted in the higher level of retained protein and slightly higher level of residual sugars in that treatment, the better digestibility of the neutral detergent fibre (NDF) and the acid detergent fibre (ADF), the lower pH and evolution of ammonia compared to the control; The fall in pH was most rapid in the SA treated silage in the first 4 days of ensilage, when it matters most for the conservation of protein, compared with the control silage and this was linked more to both lactic and total acids. Both volatile fatty acid (VFA) and lactate content increased with ensiling time, increase degree reached up near 100%; Lactate content in treatment silage was higher than that in the control, and this difference was all maintained at the level of approximately 7% - 13% almost in all cases, at length, up 16% in the case of day 15; pH value decreased with ensiling time. Moreover, pH value in most of treatment were lower than that in the control ; The digestibility of both dry matter (DM) and crude protein (CP) was appreciably improved by the use of inoculant SA. Also, in relation to reductions, of NDF and ADF were noted. An increase in milk yield of the order of 0.9kg per head per day in favour of SA was observed. Alternatively, with an average daily feed intake of 20kg, this indicates with 1 ton of corn silage milk yield could be increased by 46.5kg.     

Development of an analytical method to determine phenolic endocrine disrupting chemicals in sewage and sludge by GC/MS

An analytical method has been developed to determine hisphenoi A and five degradation products of alkyiphenoi ethoxylates in sewage and sludge. The parameters of accelerated solvent extraction （ASE） were optimized. C18 cartridge was used to clean up the sewage and the extract of sludge. The sensitivity of the mass detector was improved by adjusting the voltage of the lens manually. The instrumental detection limit was 3.2-17.8pg. The method detection limits were 2.5-11.4 ng/L and 0.3-2.1ng/g for sewage and sludge analysis respectively. The recovery and relative standard deviation for sewage analysis were 79.2%-111.4% and 6.9% - 12.6%; for sludge analysis were 68.5% - 114.0% and 8.9%-16.0%. Finally this method was applied to determining the phenolic endocrine disrupting chemicals in sewage and sludge from a sewage treatment plant in Beijing.     

Multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers

The mutation detections of KRAS and BRAF genes are of significant importance to predict the responses to anti-cancer therapy and develop new drugs. In this paper, we developed a multi-step fluorescence resonance energy transfer (FRET) assay for multiplex detection of KRAS and BRAF mutations using cationic conjugated polymers (CCP). The newly established detection system could detect as low as 2% mutant DNAs in DNA admixtures. By triggering the emission intensity change of CCP and the dyes labeled in the DNA, four possible statuses (three mutations and one wildtype) can be differentiated in one extension reaction. The detection efficiency of this new method in clinical molecular diagnosis was validated by determining KRAS and BRAF mutations of 51 formalin-fixed paraffin-embedded (FFPE) ovary tissue samples. Furthermore, the result of the CCP-based multi-step FRET assay can be directly visualized under UV light so that no expensive instruments and technical expertise are needed. Thus, the assay provides a sensitive, reliable, cost-effective and simple method for the detection of disease-related gene mutations.     

Determination of risperidone in human plasma by HPLC-MS/MS and its application to a pharmacokinetic study in Chinese volunteers

This study presents a rapid, specific and sensitive liquid chromatography/tandem mass spectrometry （LC-MS/MS） assay for determination of risperidone （RIS） in human serum using paroxetine as an internal standard （IS）. An Alltima-C18 column （2.1 mm×100 mm, 3 μm） and a mobile phase consisting of 0.1% formic acid-acetonitrile （40：60, v/v） were used for separation. The analysis was performed by selected reaction monitoring （SRM） method, and the peak area of the m/z 411.3→191.1 transition for RIS was measured versus that of the m/z 330.1→192.1 transition for IS to generate the standard curves. The assay linearity of RIS was confirmed over the range 0.25-50.00 ng/ml and the limit of quantitation was 0.05 ng/ml. The linear range corresponds well with the serum concentrations of the analytes obtained in clinical pharmacokinetic studies. Intraday and interday relative standard deviations were 1.85%-9.09% and 1.56%-4.38%, respectively. The recovery of RIS from serum was in the range of 70.20%-84.50%. The method was successfully applied to investigate the bioequivalence between two kinds of tablets （test versus reference products） in 18 healthy male Chinese volunteers. The result suggests that two formulations are bioequivalent.     

Integration of rolling circle amplification and cationic conjugated polymer for the homogeneous detection of single nucleotide polymorphisms

A novel, homogeneous and sensitive assay for the detection of single nucleotide polymorphisms (SNPs) by integration of rolling circle amplification (RCA) and cationic conjugated polymer (CCP) has been developed and tested. Mutant DNA serves as the template for specifically circularizing a padlock probe (PLP) with a sequence that is complementary to the mutant DNA. Afterwards, the mutant DNA directly acts as the primer to initiate the RCA reaction in the presence of phi29 DNA polymerase that generates a long, tandem single-strand DNA product. During the RCA reaction, fluorescein-labeled dUTPs are incorporated into the RCA products. When the CCP is introduced, efficient FRET from CCP to fluorescein occurs as a result of the strong electrostatic interactions between the CCP and the DNA produced by RCA. The wild-type DNA contains a single base mismatch with PLP with the result that the PLP is not circularized, RCA is not triggered and inefficient FRET results. By measuring the change of the emission intensities of CCP and fluorescein, it was possible to detect the SNP in a homogeneous manner. The method is sensitive and specific enough to detect 0.1 pmol/L mutant DNA and to determine a mutant allele frequency as low as 2.0%.     

Applications of two electric arc plasma torches for the beneficiation of natural quartz

Experimental beneficiation processes of quartz concentrate in arc plasma torches of two different types and electric powers were studied. An emission scanning electron microscope and a universal laser analyzer were used to investigate the structures as well as the size distributions of grains and microparticles. Inductively coupled plasma–mass spectrometry was used to determine the chemical compositions of nonstructural solid-phase mineral impurities in quartz concentrate. Results related to the modified grains' structure and size distribution, the compositions of impurities, and the gas–liquid inclusions in the quartz concentrate were investigated. The total impurities concentrations in the processed grains were found to satisfy the IOTA-STD standard(industry standard for grading high quality fused quartz products). The optimal condition(i.e., the optimal specific plasma enthalpy) for the production of high-purity quartz in arc plasma torches was found to depend on the geological-genetic type and the structural and textural features(i.e., chemical composition and gas–liquid inclusions) of the quartz concentrate.     

Detection of immunoglobulin E using an aptamer based dot-blot assay

A novel aptamer based dot-blot assay for the detection of immunoglobulin E (IgE) was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose membrane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3’,5,5’-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol/L to 1 μmol/L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol/L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.     

Determination of Polysaccharide in Radix pseudostellariae Extract by Size-Exclusion High-Performance Liquid Chromatography

A simple size-exclusion high-performance liquid chromatographic method was developed for rapid molecular mass screening to determine the average molecular mass, polydispersity, and quantity of Taizishen polysaccharides extract. The screening used a TOSOH TSK-GEL G3000SW high-performance liquid chromatographic column with 0.1 mol/L NaNO3 buffer as the mobile phase and refractive index detec- tor. The molecular mass calibration curve was linear for polysaccharide standards from 10 to 100 kDa with a correlation coefficient of 0.9991. The method can be used to analyze the quantity, average molecular mass, and polydispersity of polysaccharides. In addition, the screening method is suitable for quality control of polysaccharide preparations in Chinese medicines and pharmaceutical products.     

Time-resolved fluoroimmunoassay for quantitative determination of tylosin and tilmicosin in edible animal tissues

To quantitatively determine tylosin and tilmicosin in edible animal tissues,a time-resolved fluoroimmunoassay(TRFIA) has been developed and validated.For this purpose,desmycosin-O-carboxymethoxylamine-BSA was fixed onto microtiter plates,standards and samples were loaded and,finally,diluted europium-labeled anti-tylosin antibodies were added.Results show that the limit of detection for tylosin was 0.03 ng mL-1 and that for tilmicosin was 0.05 ng mL-1.The recoveries were 73.6% to 120.5%,with coefficients of variation below 15.6% in various biological matrices spiked with tylosin and tilmicosin at concentrations of 50-200 ngg-1.There was good correlation(R2>0.99) between the TRFIA,an enzyme-linked immunosorbent assay and high performance liquid chromatography data.In conclusion,the new TRFIA is applicable to the detection of tylosin and tilmicosin and is an effective and economical method that will enable high-throughput sample screening.The method is expected to be widely applicable.     

Rod-shaped Au@PtCu nanostructures with enhanced peroxidase-like activity and their ELISA application

Pt and its based alloy nanoparticles （NPs） have been reported to demonstrate novel enzyme-like activities. Varying composition is very important to realize the opti- mization of their functions through the tuning of electronic structure. In this paper, our effort is focused in this direction by tailoring the electronic structure of Pt NPs via alloying with copper. Using gold nanorod （Au NR） as core, a simple method to prepare PtCu alloy shell is developed （termed as Au@PtCu NR）. The introduction of copper could result in endcap-preferred growth mode owing to the lattice mismatch between alloy shell and the Au core. The variation in the electronic structure changes the substrate affinity, and enhanced affinity was found for H202. Besides, the designed Au@PtCu nanostructures have realized spatial separation of catalytic and recognition sites. Binding of recognition antibodies had negligible effect on their catalytic activity. Based on their peroxidase- like activity, a highly sensitive detection of human immunoglobulin G （IgG） was demonstrated in a direct enzyme-linked immunosorbent assay （ELISA） mode. The detection limit can be as low as 90 pg/mL.     

Novel protein detection method based on proximity-dependent polymerase reaction and aptamers

In recent years, specific detection of proteins is one of the hot issues about aptamers in proteomics. Here we reported a simple, sensitive and specific proximity-dependent protein assay with dual DNA aptamers. Thrombin was used as the model protein, and two aptamer probes with complementary sequence at 3′-end were designed for the two distinct epitopes of the protein. Association of the two aptamers with thrombin resulted in stable hybrids due to the proximity of 3′-end, then polymerase reaction was induced. The amount of obtained dsDNA was indicated using the fluorescence dye Sybr Green I. The results showed that the initial velocity of polymerase reaction had a positive correlation with concentration of thrombin. The advantages of this dual-aptamer-based approach included simple and flexible design of aptamer probes, high selectivity and high sensitivity. The detection limit was 6.9 pmol/L.     

梯度温度轧制对Q500厚板裂纹愈合的促进作用

To ensure the quality of heavy plate products as determined by ultrasonic inspection, it is necessary to effectively control defects such as cracks and shrinkage cavities in heavy plates. Generally, some defects such as large size cracks exist due to insufficient deformation in the center of traditionally rolled plates. Compared with the traditional rolling process, gradient temperature rolling (GTR) process can effectively increase deformation inside heavy plates. In this study, the effect of GTR on crack healing was analyzed through a comparison experiment with the uniform temperature rolling (UTR). The results show that the GTR process could increase the plastic strain inside the heavy plate and effectively promote the healing process of the preset cracks. The degrees of crack healing at the center and quarter thickness position of the steel plate via GTR were greater than twice those of the plate via UTR. The GTR process can significantly reduce the internal defects of heavy plates and improve the defect detection level of heavy plate products. Also, The GTR process results in the formation of new crystal grains in the crack region, which is crucial to crack healing.     

Physicochemical characteristics of various milk samples available in Pakistan

We report physicochemical characteristics of various kinds of liquid milk commercially available in Pakistan in comparison with those of fresh natural milk from animals. Milk samples were collected from local markets at Peshawar, Pakistan, and analyzed for their physical features, including moisture, total solids, specific gravity, conductivity, viscosity and titratable acidity （lactic acid equivalent）, and chemical components and macro-minerals, including total protein, casein, lactose, ash and minerals （Na, K and Mg）. These items were compared with the physicochemical characteristics of the fresh natural milk samples from buffalo, cow and goat. The results were also compared with reported nutritional quality of milk from various countries and World Health Organization （WHO） standards. We found that all the physical features and chemical components of commercially available milk in Pakistan markets meet WHO＇s requirements, except for Na, K, Ca and Mg, which are below the standards.     

Immune response in cattle inoculated with the recombinant complete polyprotein of foot-and-mouth disease virus from Bombyx mori larvae

The intact open reading frame (ORF) of foot-and-mouth disease virus (FMDV) Asia I/XJ strain was am- plified by RT-PCR and inserted into the transfer vector pVL1393 to generate plasmid pVL-ORF. Bm-N cells were transfected with pVL-ORF and linearized Bm-BacPAK6 DNA, and the recombinant silkworm baculovirus Bm-ORF containing the full ORF of FMDV was obtained. The results of indirect im- munofluorescence assay (IFA) showed that Bm-ORF could be expressed efficiently in Bm-N cell. After inoculating the early 5th instar larvae of silkworm, the polyprotein of FMDV could be detected by sandwich ELISA and empty capsid-like particles could be observed under the electron microscope. Expression products from silkworm were used as the antigen to immunize the cattle. The specific an- tibody was induced in all vaccinated animals. The immunized cattle were challenged with the virulent FMDV Asia I/XJ strain, two of the four cattle were completely protected and clinical symptoms were alleviated and delayed in the others. The results suggest that this strategy might be used to develop the new subunit FMDV vaccine.     

MGB probe assay for rapid detection of mtDNAl1778 mutation in the Chinese LHON patients by real-time PCR

Objective： Leber＇s hereditary optic neuropathy （LHON） is a maternally inherited degeneration of the optic nerve caused by point mutations of mitochondrial DNA （mtDNA）. Many unsolved questions regarding the penetrance and pathophysiological mechanism of LHON demand efficient and reliable mutation testing. This study aims to develop a minor groove binder （MGB） probe assay for rapid detection of mtDNA11778 mutation and heteroplasmy in Chinese LHON patients by real-time polymerase chain reaction （PCR）. Methods： Forty-eight patients suspected of having LHON and their maternal relatives underwent a molecular genetic evaluation, with 20 normal individuals as a control group at the same time. A real-time PCR involving two MGB probes was used to detect the mtDNA 1 1778 mutation and heteroplasmy. A linear standard curve was obtained by pUCmLHONG and pUCmLHONA clones. Results： All 48 LHON patients and their maternal relatives were positive for rntDNA11778 mutation in our assay, 27 heteroplasmic and 21 homoplasmic. Eighteen cases did not show an occurrence of the disease, while 9 developed the disease among the 27 heteroplasmic mutation cases. Eleven did not show an occurrence of the disease, while 10 cases developed the disease among 21 homoplasmic mutation cases. There was a significant difference in the incidence between the heteroplasmic and the homoplasmic mutation types. The time needed for running a real-time PCR assay was only 80 min. Conclusion： This real-time PCR assay is a rapid, reliable method for mtDNA mutation detection as well as heteroplasmy quantification. Detecting this ratio is very important for predicting phenotypic expression of unaffected carriers.     

Expression of biologically active human clotting factor Ⅸ(hFⅨ) in the mammary gland of transgenic mice

The DNA of human factor Ⅸ (hFⅨ) gene vector pMCⅨm, which had been proven to be able to express in in vitro and living cells, was introduced into 586 zygotes of Kunming White Mice by positive pressure microinjection technique with manual operation. The 499 survival embryos after microinjection were then transferred into pseudopregnant recipient mice and 216 F 0 pups were born. The analysis of PCR and Southern blot hybridization showed that, of the 216, 6 (2 females and 4 males) were integrated with foreign DNA in their genomes, giving an integration frequency of 3% (6/216). Two F\-0 female transgenic mice could express hFⅨ protein in their milk and the content was over 100 ng/mL as measured with ELISA. The biological activities of hFⅨ in the milk of two F\-0 mice were 44 67% and 79 43%, respectively.     

Remote Visualization of High-Speed Rail Power Quality Monitoring

The visualization techniques were explored for power quality monitoring.And remote visualization solutions were proposed for highspeed rail power quality monitoring.Taking the Beijing-Shanghai highspeed rail power monitoring as a study case,a remote visualization client,based on our proposed solutions,was developed for high-speed rail power quality monitoring to efficiently support power quality data analysis of the electricity business.The solutions collected data from monitoring stations deployed along the high-speed rail route and visualized the data set with a variety of visualization technologies to alert the specific stations of catastrophic events.The proposed solutions have been proved to be effective in supporting decision-making for the railway power scheduling and providing diagnosis information for quickly spotting any possible runtime failure in operation.     

Image and Video Quality Assessment Using Neural Network and SVM

An image and video quality assessment method was developed using neural network and support vector machines （SVM） with the peak signal to noise ratio （PSNR） and the structure similarity indexes used to describe image quality. The neural network was used to obtain the mapping functions between the objective quality assessment indexes and subjective quality assessment. The SVM was used to classify the images into different types which were accessed using different mapping functions. Video quality was assessed based on the quality of each frame in the video sequence with various weights to describe motion and scene changes in the video. The number of isolated points in the correlations of the image and video subjective and objective quality assessments was reduced by this method. Simulation results show that the method accurately accesses image quality. The monotonicity of the method for images is 6.94% higher than with the PSNR method, and the root mean square error is at least 35.90% higher than with the PSNR.     

The kinetic studies of direct methane oxidation to methanol in the plasma process

The research outlined here includes a study of methanol production from direct methane conversion by means of thermal and plasma method. The kinetic study, derived from thermal-based approach, was carried out to investigate thoroughly the possible intermediate species likely to be presented in the process. A set of plasma experiments was undertaken by using dielectric barrier discharge （DBD）, classified as non-thermal plasma, done at atmospheric pressure and room temperature. Plasma process yields more methanol than thermal process at the same methane conversion rates and methane to oxygen feed ratios. Oxidation reaction of thermal process resulted CO and CO2 as the most dominant products and the selectivity reached 19% and 68%, respectively. Moreover, more CO and less CO2 were produced in plasma process than in thermal process. The selectivity of CO and CO2 by plasma was 47% and 20%, respectively. Ethane （C2H6）was detected as the only higher hydrocarbon with a significant concentration. The concentration of ethane reached 9% of the total products in plasma process and 17% in thermal process. The maximum selectivity of methanol, the target material of this research, was 12% obtained by plasma method and less than 5% by thermal process. In some certain points, the kinetic model closely matched with the experimental results.     

Equilibrium between titanium ions and high-purity titanium electrorefining in a NaCl-KCl melt

TiCl_x (x = 2.17) was prepared by using titanium sponge to reduce the concentration of TiCl₄ in a NaCl-KCl melt under negative pressure. The as-prepared NaCl-KCl-TiCl_x melt was employed as the electrolyte, and two parallel crude titanium plates and one high-purity titanium plate were used as the anode and cathode, respectively. A series of electrochemical tests were performed to investigate the influence of electrolytic parameters on the current efficiency and quality of cathodic products. The results indicated that the quality of cathodic products was related to the current efficiency, which is significantly dependent on the current density and the initial concentration of titanium ions. The significance of this study is the attainment of high-purity titanium with a low oxygen content of 30×10-6.