4种植物源性成分多重real-time PCR检测方法的建立及其在食用淀粉中的应用

建立一种可同时快速检测红薯、木薯、马铃薯、玉米源性成分的多重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以红薯g3pdh基因、木薯g3pdh基因、马铃薯UGPase基因、玉米zSSIIb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,开展方法学验证,并对不同掺入比例模拟样品和实际淀粉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。与15种非目标源性均无交叉反应;对目标DNA的检测灵敏度可达到3×10^(-3) ng/μL,且具有良好的线性关系和扩增效率;对淀粉样品的检出限可达0.1%,对50份实际样品进行检测,结果与参比方法一致,说明建立的多重real-time PCR法可用于食用淀粉种类掺假鉴别检测。

Distribution, residence time, autotrophic production, and heterotrophic removal of DOP in the Mirs Bay, northern South China Sea

The importance of dissolved organic phosphorus(DOP) as a potential nutrient source for primary producers in marine systems has been recognized for up to eight decades, but currently, the understanding of the biogeochemistry of DOP is in its infancy. In the present study, monthly data between 2000 and 2014 were used to analyze the temporal and spatial distributions of DOP in the Mir Bay, the northern South China Sea. The DOP residence time(TDOP) was also investigated using a simple regression analysis in combination with chlorophyll a(Chl a) measurements while excess DOP(ΔDOP), produced by the biogeochemical processes of autotrophic production and heterotrophic removal, was determined using a two-component mixing mass-balance model in combination with salinity measurements. The results showed that the DOP concentration was(0.017±0.010) mg/L higher in the surface-water compared with the bottom-water and higher in the inner Tolo Harbour and waters adjacent to Shatoujiao compared with the main zone of the bay. Although seasonal changes and annual variability in the DOP were small, the surface DOP concentration was higher in the wet season(April–September)than in the dry season(October–March) due to the impacts of seaward discharges and atmospheric deposition into the bay. Measurement and regression results showed that the DOP release rate from phytoplankton production was about 1.83(gP)/(gChl a) and the TDOP was about 7 d, which implied that the DOP cycle in the bay was rapid. The ΔDOP was calculated from the model to be about 0.000 mg/L in the main zone of the bay and about 0.002 mg/L in the inner Tolo Harbour and waters adjacent to Shaotoujiao, suggesting that the autotrophic production of DOP was almost balanced by the heterotrophic removal in the main zone of the bay and dominated in the inner Tolo Harbour and waters adjacent to Shaotoujiao. In conclusion, the Mirs Bay is very productive and fairly heterotrophic.

马乳酒样乳杆菌马乳酒样亚种real-time PCR检测方法的建立与应用

本研究建立了一种特异性实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)检测方法,根据模式菌株马乳酒样乳杆菌马乳酒样亚种ZW3的16S rDNA序列和全基因组序列设计筛选特异性引物,采用SYBR Green I荧光染料建立real-time PCR方法,并对方法的特异性、灵敏度、重复性和混合体系等进行检测。结果表明,本研究所建立的方法特异性强、灵敏度高、重复性好,建立real-time PCR的标准曲线,其决定系数R2为0.965,具有良好的线性关系,且在马乳酒样乳杆菌马乳酒样亚种及混合体系中可以特异性检出。综上,本研究建立的real-time PCR法可以快速、准确地检测马乳酒样乳杆菌马乳酒样亚种,为马乳酒样乳杆菌的特异性定性定量检测提供了一种新的方法。

Turing Instability of Gray-Scott Reaction-Diffusion Model with Time Delay Effects

The reaction diffusion Gray-Scott model with time delay is put forward with the assumption of Neumann boundary condition is satisfied. Based on the Turing bifurcation condition, the Turing curves on two parameter plane are discussed without time delay. The normal form is computed via applying Lyapunov-Schmidt reduction method in system of PDE, and the bifurcating direction of pitchfork bifurcation underlying codimension-1 singularity of Turing point is computed. The continuation of Pitchfork bifurcation is simulated with varying free parameter continuously near the turing point, which is in coincidence with the theoritical analysis results. The wave pattern formation in the case of turing instability is also simulated which discover Turing oscillation phenomena from periodicity to irregularity.

Applying real-time control to enhance the performance of nitrogen removal in CAST system

A bench-scale reactor（72 L） red with domestic sewage, was operated more than 3 months with three operation modes： traditional mode, modified mode and real-time control mode, so as to evaluate effects of the operation mode on the system performance and to develop a feasible control strategy. Results obtained from fixed-time control study indicate that the variations of the pH and oxidation-reduction potential（ORP） profiles can represent dynamic characteristics of system and the cycle sequences can be controlled and optimized by the control points on the pH and ORP profiles. A control strategy was, therefore, developed and applied to real-time control mode. Compared with traditional mode, the total nitrogen（TN） removal can be increased by approximately 16% in modified mode and a mean TN removal of 92% was achieved in real-time control mode. Moreover, approximately 12.5% aeration energy was saved in real- time control mode. The result of this study shows that the performance of nitrogen removal was enhanced in modified operation mode. Moreover, the real-time control made it possible to optimize process operation and save aeration energy.

4种动物源性成分多重real-time PCR检测方法的建立及其在驴肉制品检测中的应用

建立一种同时快速检测驴、马、猪及鸭源性成分的四重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以4种源性成分的Nad5、ATpase6、ATP8、cytb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,并对该方法进行方法学验证,同时对不同掺入比例模拟样品、不同加工工艺模拟样品和实际驴肉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。当Ct值≤35.0时,方法对16种非目标源性具有良好特异性;灵敏度可检测到质量浓度为2×10^(-4)ng/μL的模板DNA;对生肉的检出限为肉含量的0.001%,对熟肉制品的检出限为肉含量的0.01%;对100份实际样品进行检测,结果与标准方法一致,说明建立的多重real-time PCR法可用于肉及肉制品中常见掺假源性成分的检测。

Statistical analysis of generalized exponential distribution under progressive censoring with binomial removals

The estimation of generalized exponential distribution based on progressive censoring with binomial removals is presented, where the number of units removed at each failure time follows a binomial distribution. Maximum likelihood estimators of the parameters and their confidence intervals are derived. The expected time required to complete the life test under this censoring scheme is investigated. Finally, the numerical examples are given to illustrate some theoretical results by means of Monte-Carlo simulation.

Removal of nitrogen and phosphorus in a combined A^2/O-BAF system with a short aerobic SRT

A bench-scale anaerobic/anoxic/aerobic process-biological aerated filter （A^2/O-BAF） combined system was carded out to treat wastewater with lower C/N and C/P ratios. The A^2/O process was operated in a short aerobic sludge retention time （SRT） for organic pollutants and phosphorus removal, and denitrification. The subsequent BAF process was mainly used for nitrification. The BAF effluent was partially returned to anoxic zone of the A^2/O process to provide electron acceptors for denitrification and anoxic P uptake. This unique system formed an environment for reproducing the denitdfying phosphate-accumulating organisms （DPAOs）. The ratio of DPAOs to phosphorus accumulating organisms （PAOs） could be maintained at 28% by optimizing the organic loads in the anaerobic zone and the nitrate loads into the anoxic zone in the A^2/O process. The aerobic phosphorus over-uptake and discharge of excess activated sludge was the main mechanism of phosphorus removal in the combined system. The aerobic SRT of the A^2/O process should meet the demands for the development of aerobic PAOs and the restraint on the nitrifiers growth, and the contact time in the aerobic zone of the A^2/O process should be longer than 30 min, which ensured efficient phosphorus removal in the combined system. The adequate BAF effluent return rates should be controlled with 1--4 mg/L nitrate nitrogen in the anoxic zone effluent of A^2/O process to achieve the optimal nitrogen and phosphorus removal efficiencies.

Development and application of a real-time polymerase chain reaction method for Campylobacter jejuni detection

AIM:To develop a real-time polymerase chain reaction(PCR) method to detect and quantify Campylobacter jejuni(C.jejuni) from stool specimens.METHODS:Primers and a probe for real-time PCR were designed based on the specific DNA sequence of the hipO gene in C.jejuni.The specificity of the primers and probe were tested against a set of Campylobacter spp.and other enteric pathogens.The optimal PCR conditions were determined by testing a series of conditions with standard a C.jejuni template.The detection limits were obtained using purified DNA from bacterial culture and extracted DNA from the stool specimen.Two hundred and forty-two specimens were analyzed for the presence of C.jejuni by direct bacterial culture and real-time PCR.RESULTS:The optimal PCR system was determined using reference DNA templates,1 × uracil-DNA glycosylase,3.5 mmol/L MgCl 2,1.25 U platinum Taq polymerase,0.4 mmol/L PCR nucleotide mix,0.48 μmol/L of each primer,0.2 μmol/L of probe and 2 μL of DNA template in a final volume of 25 μL.The PCR reaction was carried as follows:95 ℃ for 4 min,followed by 45 cycles of 10 s at 95 ℃ and 30 s at 59 ℃.The detection limit was 4.3 CFU/mL using purified DNA from bacterial culture and 10 3 CFU/g using DNA from stool specimens.Twenty(8.3%,20/242) C.jejuni strains were isolated from bacterial culture,while 41(16.9%,41/242) samples were found to be positive by realtime PCR.DNA sequencing of the PCR product indicated the presence of C.jejuni in the specimen.One mixed infection of C.jejuni and Salmonella was detected in one specimen and the PCR test for this specimen was positive.CONCLUSION:The sensitivity of detection of C.jejuni from stool specimens was much higher using this PCR assay than using the direct culture method.

Background removal and weld defect detection based on energy distribution of image

The lateral wave in ultrasonic TOFD （time of flight diffraction） image has a tail in transit time, which disturbs the detection and evaluation of shallow weld defect. Meanwhile, the lateral wave and back-wall echo that act as background add redundant data in digital image processing. In order to separate defect wave from lateral wave and prepare the way for following image processing, an algorithm of background removal method named as mean-subtraction is developed. Based on this, an improved method by statistic of the energy distribution in the image is proposed. The results show that by choosing proper threshold value according to the axial energy distribution of the image, the background can be removed automatically and the defect section becomes predominant. Meanwhile, diffractive wave of shallow weld defect can be separated from lateral wave effectively.

Kinetic Model of Hydrogenation for Removal of Trace Olefins from Alkylation Mixture Formed during Linear Alkylbenzene Synthesis

Through the hydrogenation experiments for removal of olefin impurities in effluent mixture obtained from alkylation reactor for linear alkylbenzene synthesis, the reaction kinetics was studied. The experimental results showed that the catalytic activity of Pd/Al2 O3 among four kinds of catalysts was the highest, and the olefin conversion reached 100%when the reaction temperature was higher than 140 ℃. The kinetic model was established through parameters estimation.The experimental validation of the kinetic model showed that the kinetic model had higher simulation accuracy. The results of prediction by the kinetic model indicated that with the intensification of reaction conditions, the olefin conversion would increase and the bromine index of the linear alkyl benzene would decrease. Under the conditions covering a WHSV of 2.0 h^(-1),a temperature of 90 ℃, and a hydrogen partial pressure of 1.5 MPa, the olefin conversion was 99.67%, and the bromine index of the linear alkyl benzene could be reduced to less than 20.00 mgBr/(100 g).

Effect of infiltrating time on interfacial reaction and properties of tungsten particles reinforced Zr-based bulk metallic glass composites

The tungsten particles reinforced Zr41.2Ti13.8Cu12.5Ni10Be22.5 (Vit 1 alloy) bulk metallic glass composites (BMGCs) were prepared by the melt infiltrating casting method with the infiltrating time of 1, 5 and 10 min, respectively. The changes of interfacial reaction and compression properties of the bulk metallic glass composites with different infiltrating times were studied. Results show that with the increase of infiltrating time, tiny nanocrystals are generated at the interfacial boundary of tungsten particles and the amorphous matrix, and the size of tiny crystals increases with the infiltrating time. When the infiltrating time is 10 min, polygonal crystals with a larger size are also generated within the amorphous matrix. The compressive strength of the composites also increases with the infiltrating time. When the infiltrating time is 10 min, the compressive strength of the composite reaches 2,030 MPa and the compression strain is 44%. The fracture morphology of the composite materials is in a vein-like pattern and the melting phenomenon is found on the fracture surface. In addition, the density of the shear bands during the compressive tests of the composite materials increases with the infiltrating time.

Singularly perturbed reaction diffusion equations with time delay

A class of initial boundary value problems of differential-difference equations for reaction diffusion with a small time delay is considered. Under suitable conditions and by using the stretched variable method, a formal asymptotic solution is constructed. Then, by use of the theory of differential inequalities, the uniform validity of the solution is proved.

Influence of Reaction Time on Growth Behaviours of Mg-Al LDH Films

Mg-Al LDH film was fabricated on anodized magnesium alloy AZ31 by in-situ growth method.The characteristics of the film were investigated by X-ray diffraction(XRD),Fourier transform infrared(FT-IR),scanning electron microscopy(SEM)observation and electrochemical tests.The results showed that the crystallinity of LDH film was increased with reaction time.When the reaction time was more than 12 h,the LDH film had complete crystal layered structure.The anodic oxide film was sealed basically by the growth of LDH nanosheets after 6 h reaction.The corrosion resistance of the films became better with the increasing of reaction time.However,after 12 h reaction,the corrosion resistance of the film decreased.The formation behavior of the Mg-Al LDH film was proposed.That the competitive growth of LDH grains,which contains dissolution and recrystallization,was proposed to explain the change of the growth and corrosion resistance of the Mg-Al LDH film.

A statistical inference for generalized Rayleigh model under Type-Ⅱ progressive censoring with binomial removals

This paper considers the parameters and reliability characteristics estimation problem of the generalized Rayleigh distribution under progressively Type-Ⅱ censoring with random removals,that is,the number of units removed at each failure time follows the binomial distribution.The maximum likelihood estimation and the Bayesian estimation are derived.In the meanwhile,through a great quantity of Monte Carlo simulation experiments we have studied different hyperparameters as well as symmetric and asymmetric loss functions in the Bayesian estimation procedure.A real industrial case is presented to justify and illustrate the proposed methods.We also investigate the expected experimentation time and discuss the influence of the parameters on the termination point to complete the censoring test.

Mechanisms of shale hydration and water block removal

Shale samples of Silurian Longmaxi Formation in the Changning area of the Sichuan Basin, SW China, were selected to carry out scanning electron microscopy, CT imaging, high-pressure mercury injection, low-temperature nitrogen adsorption and imbibition experiments to compare the hydration characteristics of montmorillonite and illite, analyze the main factors affecting the water block removal of shale, and reveal the mechanisms of pore structure evolution during shale hydration. The hydration characteristics of shale are closely related to the composition of clay minerals, the shale with high illite content is not susceptible to hydration and thus has limited room for pore structure improvement;the shale with high montmorillonite is susceptible to hydration expansion and thus has higher potential of pore structure improvement by stimulation;the shale with high illite content has stronger imbibition in the initial stage, but insufficient diffusion ability, and thus is likely to have water block;the shale with high montmorillonite content has weaker imbibition in the initial stage but better water diffusion, so water blocking in this kind of shale can be removed to some degree;the shale reservoir has an optimal hydration time, when it is best in physical properties, but hydration time too long would cause damage to the reservoir, and the shale with high illite content has a shorter optimal hydration time;inorganic cations can inhibit the hydration of clay minerals and have stronger inhibition to illite expansion, especially K^(+);for the reservoir with high content of montmorillonite, the cation content of fracturing fluid can be lowered to promote the shale hydration;fracturing fluid with high K^(+) content can be injected into reservoirs with high illite content to suppress hydration.

Evaluation of whole body reaction time and one leg with eye closed balance in elderly Japanese

We evaluated whole body reaction time and one leg with eye closed balance in elderly Japanese. A total of 2059 elderly Japanese (730 men and 1329 women), aged 60 - 79 years, were enrolled in the cross-sectional investigation study. Anthropometric parameters, whole body reaction time and one leg with eye closed balance were measured. Whole body reaction time was significantly higher and one leg with eye closed balance was significantly lower in 70’s than those in 60’s in both sexes. Two hundred twenty seven men (31.1%) and 533 women (40.1%) were having no medications. In men, whole body reaction time was significantly higher and one leg with eye closed balance was significantly lower in subjects with medications than those in subjects without in 60’s. However, significant differences of whole body reaction time and one leg with eye closed balance between subjects with and without medications in men with 70’s and women with 60’s and 70’s were not noted. This standard mean value may provide a useful reference database for evaluating whole body reaction time and one leg with eye closed balance in Japanese elderly subjects.

Changes in reaction time, coefficient of variance of reaction time, and autonomic nerve function in the mental fatigue state caused by long-term computerized Kraepelin test workload in healthy volunteers

Fatigue is a common sense caused by crushing labor, stressful social events and various illnesses. It is usually judged by their subjective symptoms, but it should be evaluated in an objective perspective. Here we show that the decrease of working efficiency and sympathetic hyperactivity are associated with mental fatigue state caused by prolonged mental workload. Recently we made a new mental fatigue model of healthy volunteers caused by long-term computerized Kraepelin test (CKT) workload. CKT is our new software for automatically checking the calculation capability, with which it is easy to determine the reaction time (RT), coefficient of variance of reaction time (CV), and accuracy of the answers (AC) during tasks. We put 24 healthy volunteers into the fatigue state by subjecting them to 120 minutes’ CKT workload, and then studied the changes in fatigue sensation, RT, CV, and AC before and after the CKT workload. The fatigue sensation, RT, and CV were clearly increased by the fatigue-inducing task and recovered during the resting period. We also studied the changes in autonomic nerve activity by using heart rate variability analysis. The low/high frequency component ratio (LF/HF) was signifi-cantly increased by the fatigue-inducing task and decreased by resting, suggesting that mental stress causes a relatively sympathetic nerve activity-dominant state. Therefore, our new fatigue model involving a long-term CKT workload is a good mental fatigue model to provide much information about the fatigue state simultane-ously, and the increase of RT, CV, and proportion of sympathetic activity (LF/HF) are associated with mental fatigue state. These might be useful objective biomarkers or evaluating a mental fatigue state.

Comparison between Auditory and Visual Simple Reaction Times

Objective: The purpose of this study was to find out whether the simple reaction time was faster for auditory or visual stimulus and the factors responsible for improving the performance of the athlete. Methodology: 14 subjects were as- signed randomly into groups consisting of 2 members. Both the members from each group performed both the visual and auditory tests. The tests were taken from the DirectRT software program from a laptop. The DirectRT software consists of Testlabvisual and Testlabsounds to test the reaction times to visual and auditory stimuli. The 2 members from each group completed both the visual and auditory reaction times, the data was taken and the mean reaction time was calculated excluding the first and last values. Results: The results show that the mean visual reaction time is around 331 milliseconds as compared to the mean auditory reaction time of around 284 milliseconds. Conclusion: This shows that the auditory reaction time is faster than the visual reaction time. And also males have faster reaction times when compared to females for both auditory as well as visual stimuli.