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。

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.

垂体后叶粉中3种动物源性成分多重real-time PCR检测方法的建立及应用

目的基于三重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)技术同时快速检测垂体后叶粉中的猪、牛、羊性动物源性成分。方法采用CTAB法提取样品及对照品DNA,以线粒体基因为靶基因设计并合成三重real-time PCR的引物探针,并对建立方法进行验证,用于检测垂体后叶粉样品是否与标识一致。结果猪、牛、羊源性成分的最低检测浓度为10 pg/μL,采用建立的方法检测3批垂体后叶粉样品,均只检测出猪源性成分。结论建立的方法可用于同时检测垂体后叶粉中的猪、牛、羊源性成分,可为相关工作提供技术支持。

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。

Changing the balance of the MTO reaction dual-cycle mechanism: Reactions over ZSM-5 with varying contact times

The methanol to olefins （MTO） reaction was performed over ZSM‐5 zeolite at 300℃ under various methanol weight hourly space velocity （WHSV） values. During these trials, the catalytic perfor‐mance was assessed, in addition to the formation and function of organic compounds retained in the zeolite. Analysis of reaction effluents and confined organics demonstrated a dual‐cycle reaction mechanism when employing ZSM‐5. The extent of the hydrogen transfer reaction, a secondary reac‐tion in the MTO process, varied as the catalyst‐methanol contact time was changed. In addition, 12C/13C‐methanol switch experiments indicated a relationship between the dual‐cycle mechanism and the extent of the hydrogen transfer reaction. Reactions employing a low methanol WHSV in conjunction with a long contact time favored the hydrogen transfer reaction to give alkene products and promoted the generation and accumulation of retained organic species, such as aromatics and methylcyclopentadienes, which enhance the aromatic cycle. When using higher WHSV values, the reduced contact times lessened the extent of the hydrogen transfer reaction and limited the genera‐tion of methylcyclopentadienes and aromatic species. This suppressed the aromatic cycle, such that the alkene cycle became the dominant route during the MTO reaction.

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.

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.

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.

Detection of the Pandemic H1N1/2009 Influenza A Virus by a Highly Sensitive Quantitative Real-time Reverse-transcription Polymerase Chain Reaction Assay

A quantitative real time reverse-transcription polymerase chain reaction （qRT-PCR） assay with specific primers recommended by the World Health Organization （WHO） has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin （HA） genes of the pandemic HlN1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold more sensitive than those with the WHO-recommended primers as the detection limits of the assays with our primers and the WHO-recommended primers were 2.5 and 25 copies of target RNA per reaction, respectively. When tested with 83 clinical samples, 32 were detected to be positive using the qRT-PCR assays with our designed primers, while only 25 were positive by the assays with the WHO-recommended primers. These results suggest that the qRT-PCR system with the newly designed primers represent a highly sensitive assay for diagnosis of the pandemic HIN1/2009 virus infection.

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.

Dynamic detection of wake-sleep transition with reaction time-magnitude

BACKGROUND： According to observable behaviors, sleep and wakefulness are two fundamentally different behavioral states. Although electroencephalogram （EEG） is traditionally used to define sleep stage, it is difficult to detect or to quantify microarousals or disruptions during sleep. In addition initial sleep cannot be defined. It is thought that the wake-sleep transition cannot be defined by EEG patterns. OBJECTIVE： To observe the behavioral response magnitude during wake-sleep transition by EEG monitoring and to define the wake-sleep transition. DESIGN, TIME AND SETTING： A behavioral and neural network study was performed at the Key Lab of Human Being Development and Mental Health of Central China Normal University, and Lab of Brain and Cognitive Science of South Central University for Nationalities, China in July 2007. PARTICIPANTS： A total of 30 healthy volunteers, of equal gender and aged （19.7 ± 1.1 ） years, were recruited from the Central China Normal University, China for this study. None of the subjects had undergone EEG recording prior to this study or received any medication for sleep disturbances. METHODS： A novel adaptive approach was applied to detect wake-sleep transition, which avoided stimulus-induced waking. To test the difference between wake state and wake-sleep transition, the amount of self-information and mutual-information were effective parameters to analyze wake-sleep transition. MAIN OUTCOME MEASURES： The following parameters were measured： morphological changes in reaction time-magnitude, as well as correlation between phase changes and sleep, and wake and wake-sleep transition. RESULTS： There were three typical phases in morphological changes of reaction time-magnitude. With regard to the behavioral definition and criterion for sleep, the phase morphological characteristics displayed good correlation with behavioral states, such as sleep, wakefulness, and sleep onset. Entropy as an indicator of brain cognitive processes was introduced to test for differences between the wakefulness and sleep onset phase. Results indicated a cognitive declined transitional period different between sleep and wake. After staggered cognitive changes during the wake-sleep transition, the brain underwent marked alterations and transitioned into sleep quickly with no bi-directional EEG changes. CONCLUSION： Wake-sleep transition exists as an independent stage.

A Study of Visual Event-Related Potential and Reaction Time in Elderly People: Comparative Analysis of the Scores of Intelligence Test in 30 Subjects

The results of visual event-related potential(ERP)examinations and reactiontime(RT)tests were reported in 30 elders and compared with their performanceintellegence quotient(PIQ)scores.The subjects consisted of 18 males and 12 femalesaged 50-71(mean 61.4)years old.No history of central nervous system disease wasfound.The visual stimuli were randomly presented to the subject,including three sym-bols:E as target stimulus with 0.15 probability,and H and E as nontarget stimuliwith 0.15 and 0.70 probability respectively.The recording electrodes were placed on Fzand Pz.The duration from the subject seeing the target to touching a button immediatelywas considered as reaction time(RT).It was shown that the P3 latency at Pz was longer than that at Fz and the P3amplitude at Pz was larger than that at Fz,and that the RT was longer than P3 latencywith obvious effect of distribution(P【0.05 at Fz and P】0.05 at Pz)as well .The higherthe PIQ scores,the longer the RT and the P3 latency.It is suggested that the ERPmight reflect the differences of PIQ scores,and the P3 is an objective index.We considerthat the research of ERP is of great interest in the neuropsychological and neurological sci-ences.

Changes in brain activation patterns according to cross-training effect in serial reaction time task An functional MRI study

Cross-training is a phenomenon related to motor learning, where motor performance of the untrained limb shows improvement in strength and skill execution following unilateral training of the homologous contralateral limb. We used functional MRI to investigate whether motor performance of the untrained limb could be improved using a serial reaction time task according to motor sequential learning of the trained limb, and whether these skill acquisitions led to changes in brain activation patterns. We recruited 20 right-handed healthy subjects, who were randomly allocated into training and control groups. The training group was trained in performance of a serial reaction time task using their non-dominant left hand, 40 minutes per day, for 10 days, over a period of 2 weeks. The control group did not receive training. Measurements of response time and percentile of response accuracy were performed twice during pre- and post-training, while brain functional MRI was scanned during performance of the serial reaction time task using the untrained right hand. In the training group, prominent changes in response time and percentile of response accuracy were observed in both the untrained right hand and the trained left hand between pre- and post-training. The control group showed no significant changes in the untrained hand between pre- and post-training. In the training group, the activated volume of the cortical areas related to motor function （i.e., primary motor cortex, premotor area, posterior parietal cortex） showed a gradual decrease, and enhanced cerebellar activation of the vermis and the newly activated ipsilateral dentate nucleus were observed during performance of the serial reaction time task using the untrained right hand, accompanied by the cross-motor learning effect. However, no significant changes were observed in the control group. Our findings indicate that motor skills learned over the 2-week training using the trained limb were transferred to the opposite homologous limb, and motor skill acquisition of the untrained limb led to changes in brain activation patterns in the cerebral cortex and cerebellum.

How the Brain Process Stimulus-Response Conflict? New Insights from Lateralized Readiness Potentials Scalp Topography and Reaction Times

Stimulus-Response Compatibility (SRC) refers to the fact that some tasks are performed easier and better than others because of the way stimuli and responses are paired with each other. To assess the brain responses to stimulus-response conflicts, we investigated the behavioral (accuracy and Reaction Times: RTs) as well as the physiological response (Lateralized Readiness Potentials: LRP) modulations in a positional blocked and a conditional mixed design in twelve university students. Results revealed that the performance was less accurate and the RTs, as well as the LRP onset, were delayed under the mixed conditional design. A greater compatibility effect was also noted on accuracy, RTs and LRP onset latency in the mixed design. Consistent with these findings, smaller peak activation at fronto-central areas suggests that more selective inhibition is needed in a mixed design context. Despite a smaller activation, the topographical distribution is similar in both designs. These results indicate that the translation time between stimulus- and response codes are greater under the mixed instruction, while the similar LRP topography suggests that common neural structures underlie LRPs in response to both type of designs.

Luminescent properties dependence of water-soluble CdTe quantum dots on stabilizing agents and reaction time

The influence of stabilizing agents and reaction time on the luminescent properties of water-soluble CdTe quantum dots(QDs) was discussed.The thioglycolic acid(TGA)-CdTe ODs were characterized by TEM,XRD and FTIR.It is found that larger-size QDs can be synthesized more easily when L-cysteine(Cys) or golutathione(GSH) is chosen as stabilizing agent and TGA is proper to prepare highly luminescent QDs because of the effect between Cd2+ and sulfhydryl group.Furthermore,the absorption wavelength,full width at half maximum(FWHM),stokes shift,photoluminescence(PL) quantum yield and PL stability of TGA-CdTe are strongly dependent on reaction time,in which the absorption wavelength changes against reaction time with an exponential function.The TGA-CdTe QDs prepared at 2 h possess more excellent luminescent properties.

Reaction Time of Children with and without Autistic Spectrum Disorders

This study was conducted to evaluate and compare simple and choice reaction times for the comparison of perceptual-motor development levels found in children with and without autism. The participants of the study consisted of 24 children with autism and 10 typically developing (TD) children. Within the group of children with autism, only 10 were able to offer a full set data and four offered partial data. Data were collected through a computer-based procedure of reaction time test software, where the participant reacted by pressing a key on a laptop upon the appearance of one of the boxes turning yellow. Simple and choice reaction times appeared on the screen and were then recorded. The means and standard deviations were calculated for comparison and the data were evaluated from the results of a t-test. Significant differences were found among all reaction time tasks, whereas the group consisting of children with autism demonstrated slower reaction times and greater standard deviations compared with the typically developing/control group.