The Identification of Phenylalanine Ammonia-Lyase(PAL)Genes from Pinus yunnanensis and an Analysis of Enzyme Activity in vitro

Phenylalanine ammonia lyase(PAL)is the rate-limiting and pivotal enzyme of the general phenylpropanoid path-way,but few reports have been found on PAL genes in Pinus yunnanensis.In the present study,three PAL genes were cloned and identified from P.yunnanensis seedlings for thefirst time,namely,PyPAL-1,PyPAL-2,and PyPAL-3.Our results indicated that the open-reading frames of PyPAL genes were 2184,2157,and 2385 bp.Phylogenetic tree analysis revealed that PyPALs have high homology with other known PAL genes in other plants.In vitro enzymatic analysis showed that all three PyPAL recombinant proteins could catalyze the deamination of L-phenylalanine to form trans-cinnamic acid,but only PAL1 and PAL2 can catalyze the conversion of L-tyrosine toρ-coumaric acid.Three PyPAL genes were expressed in different tissues in 1-year-old P.yunnanensis,and such genes had different expression patterns.This study lays a foundation for further understanding of the biosynthesis of secondary metabolites in P.yunnanensis.

Protein profile of human hepatocarcinoma cell line SMMC-7721:Identification and functional analysis

AIM： TO investigate the protein profile of human hepatocarcinoma cell line SMMC-7721, to analyze the specific functions of abundant expressed proteins in the processes of hepatocarcinoma genesis, growth and metastasis, to identify the hepatocarcinoma-specific biomarkers for the early prediction in diagnosis, and to explore the new drug targets for liver cancer therapy. METHODS： Total proteins from human hepatocarcinoma cell line SMMC-7721 were separated by two-dimensional electrophoresis （2DE）. The silver-stained gel was analyzed by 2DE software Image Master 2D Elite. Interesting protein spots were identified by peptide mass fingerprinting based on matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and database searching. RESULTS： We obtained protein profile of human hepatocarcinoma cell line SMMC-7721. Among the twenty-one successfully identified proteins, mitofilin, endoplasmic reticulum protein ERp29, ubiquinol-cytochrome C reductase complex core protein I, peroxisomal enoyl CoA hydratase, peroxiredoxin-4 and probable 3-oxoacid CoA transferase 1 precursor were the six novel proteins identified in human hepatocarcinoma cells or tissues. Specific functions of the identified heat-shock proteins were analyzed in detail, and the results suggested that these proteins might promote tumorigenesis via inhibiting cell death induced by several cancer-related stresses or via inhibiting apoptosis at multiple points in the apoptotic signal pathway. Other identified chaperones and cancer-related proteins were also analyzed.CONCLUSION： Based on the protein profile of SMMC-7721 cells, functional analysis suggests that the identified chaperones and cancer-related proteins have their own pathways to contribute to the tumorigenesis, tumor growth and metastasis of liver cancer. Furthermore, proteomic analysis is indicated to be feasible in the cancer study.

Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batatas (L.) Lam.

This paper reported firstly successful cloning of lycopene ε-cyclase （lbLCYe） gene from sweetpotato, lpomoea batatas （L.） Lam. Using rapid amplification of cDNA ends （RACE）, lbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence oflbLCYe gene contained a 1236 bp open reading frame （ORF） encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point （pI） of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD （flavinadenine dinucleotide）/NAD（P） （nicotinamide adenine dinucleotide phosphate）-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA oflbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco （cv. Wisconsin 38） expressing [bLCYe gene accumulated significantly more ^-carotene compared to the untransformed control plants. These results showed that lbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.

Analysis of the regional spectral properties in northwestern South China Sea based on an empirical orthogonal function

This study presents an analysis of the spectral characteristics of remote sensing reflectance(Rrs)in northwestern South China Sea based on the in situ optical and water quality data for August 2018.Rrswas initially divided into four classes,classes A to D,using the max-classification algorithm,and the spectral properties of whole Rrs were characterized using the empirical orthogonal function(EOF)analysis.Subsequently,the dominant factors in each EOF mode were determined.The results indicated that more than 95%of the variances of Rrs are partly driven by the back-scattering characteristics of the suspended matter.The initial two EOF modes were well correlated with the total suspended matter and back-scattering coefficient.Furthermore,the first EOF modes of the four classes of Rrs(A-D Rrs-EOF1)significantly contributed to the total variances of each Rrs class.In addition,the correlation coefficients between the amplitude factors of class A-D Rrs-EOF1 and the variances of the relevant water quality and optical parameters were better than those of the unclassified ones.The spectral shape of class ARrs-EOF1 was governed by the absorption characteristic of chlorophyll a and colored dissolved organic matter(CDOM).The spectral shape of class B Rrs-EOF1 was governed by the absorption characteristic of CDOM since it exhibited a high correlation with the absorption coefficient of CDOM(ag(λ)),whereas the spectral shape of class C Rrs-EOF1 was governed by the back-scattering characteristics but not affected by the suspended matter.The spectral shape of class D Rrs-EOF1 exhibited a relatively good correlation with all the water quality parameters,which played a significant role in deciding its spectral shape.

The effect of water spreading system on the functionality of rangeland ecosystems

In recent decades, the control of floods is an efficient management practice for the rehabilitation of rangelands in most arid and semiarid areas. To evaluate the benefits, we used the Landscape Function Analysis （LFA） method to assess the function of patches and qualitative capability of a rangeland ecosystem in Gareh By- gone region, Fars province, southwestern Iran. Landscape functionality depends on soil, water and nutrient （col- lectively called ＂resources＂） conservation and use within a given ecosystem. Many landscapes are naturally het- erogeneous in terms of resource control and possess patches, where resources tend to accumulate, and in- ter-patches. Assessing rangeland health and function of landscape patches in response to environment and man- agement can give rise to correct management decisions for qualitative improvement of the ecosystem. Therefore, our study area was divided into two parts, i.e. water spreading and control parts, and sampling was done using LFA method in each part separately. Structural parameters, including the number, length and width of patches, and the mean length of inter-patches, were determined by the method to characterize the functional status of the monitoring sites. For each patch/inter-patch type identified in the transect organization log, we recorded its soil surface proper- ties classified according to the Soil Surface Assessment Method. The density, canopy cover and composition of plants were then assessed. The results showed that the number of ecological patches and their dimensions were significantly increased in the water spreading site. Soil stability and the values of nutrient cycling indices were in- creased but the infiltration values were decreased in the water spreading site. It could be related to the effect of suspended materials transported by floods to the soils in the study area. The improvement of ecological patches and rangeland ecosystem was achieved where water spreading systems were practiced. Therefore it can be con- cluded that water spreading as a management plan plays an important role in arid land ecosystem functionality.

Semen analysis and sperm function testing

Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization （WHO） manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external qualitY control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm-zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.

Bioinformatics Analysis of Genes and Pathways of CD11b^(+)/Ly6C^(intermediate)Macrophages after Renal Ischemia-Reperfusion Injury

Renal ischemia-reperfusion injury(IRI)is a major cause of acute kidney injury(AKI),which could induce the poor prognosis.The purpose of this study was to characterize the molecular mechanism of the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.The gene expression profiles of CD11 b^(+)/Ly6 C^(intermediate)macrophages of the sham surgery mice,and the mice 4 h,24 h and 9 days after renal IRI were downloaded from the Gene Expression Omnibus database.Analysis of m RNA expression profiles was conducted to identify differentially expressed genes(DEGs),biological processes and pathways by the series test of cluster.Protein-protein interaction network was constructed and analysed to discover the key genes.A total of 6738 DEGs were identified and assigned to 20 model profiles.DEGs in profile 13 were one of the predominant expression profiles,which are involved in immune cell chemotaxis and proliferation.Signet analysis showed that Atp5 a1,Atp5 o,Cox4 i,Cdc42,Rac2 and Nhp2 were the key genes involved in oxidation-reduction,apoptosis,migration,M1-M2 differentiation,and proliferation of macrophages.RPS18 may be an appreciate reference gene as it was stable in macrophages.The identified DEGs and their enriched pathways investigate factors that may participate in the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.Moreover,the vital gene Nhp2 may involve the polarization of macrophages,which may be a new target to affect the process of AKI.

Genome-wide Analysis of Ovate Family Proteins in Arabidopsis

Arabidopsis thaliana ovate family proteins （AtOFPs） is a newly found plant-specific protein family interacting with TALE （3-aa loop extension homeodomain proteins） homeodomain proteins in Arabidopsis. Here, based on bioinformatic analysis, we found that Arabidopsis genome actually encoded 17 OVATE domain-containing proteins. One of them, AtOFP19, has not been previously identified. Based on their amino acid sequence similarity, AtOFPs proteins can be divided into two groups. Most of the AtOFPs were located in nuclear, four of them were presented in chloroplast and the remaining two members appeared in cytoplasmic. A genome- wide microarray based gene expression analysis involving 47 stages of vegetative and reproductive development revealed that AtOFPs have diverse expression pattems. Investigation of proteins interaction showed that nine AtOFPs only interacted with TALE homeodomain proteins, which are fundamental regulators of plant meristem function and leaf development. Our work could provide important leads toward functional genomics studies of ovate family proteins, which may be involved in a previously unrecognized control mechanism in plant development

Comprehensive Reliability Allocation Method for CNC Lathes Based on Cubic Transformed Functions of Failure Mode and Effects Analysis

Reliability allocation of computerized numerical controlled（CNC）lathes is very important in industry.Traditional allocation methods only focus on high-failure rate components rather than moderate failure rate components,which is not applicable in some conditions.Aiming at solving the problem of CNC lathes reliability allocating,a comprehensive reliability allocation method based on cubic transformed functions of failure modes and effects analysis（FMEA）is presented.Firstly,conventional reliability allocation methods are introduced.Then the limitations of direct combination of comprehensive allocation method with the exponential transformed FMEA method are investigated.Subsequently,a cubic transformed function is established in order to overcome these limitations.Properties of the new transformed functions are discussed by considering the failure severity and the failure occurrence.Designers can choose appropriate transform amplitudes according to their requirements.Finally,a CNC lathe and a spindle system are used as an example to verify the new allocation method.Seven criteria are considered to compare the results of the new method with traditional methods.The allocation results indicate that the new method is more flexible than traditional methods.By employing the new cubic transformed function,the method covers a wider range of problems in CNC reliability allocation without losing the advantages of traditional methods.

Wave Energy Estimation by Using A Statistical Analysis and Wave Buoy Data near the Southern Caspian Sea

Statistical analysis was done on simultaneous wave and wind using data recorded by discus-shape wave buoy. The area is located in the southern Caspian Sea near the Anzali Port. Recorded wave data were obtained through directional spectrum wave analysis. Recorded wind direction and wind speed were obtained through the related time series as well. For 12-month measurements（May 25 2007-2008）, statistical calculations were done to specify the value of nonlinear auto-correlation of wave and wind using the probability distribution function of wave characteristics and statistical analysis in various time periods. The paper also presents and analyzes the amount of wave energy for the area mentioned on the basis of available database. Analyses showed a suitable comparison between the amounts of wave energy in different seasons. As a result, the best period for the largest amount of wave energy was known. Results showed that in the research period, the mean wave and wind auto correlation were about three hours. Among the probability distribution functions, i.e Weibull, Normal, Lognormal and Rayleigh, ＂Weibull＂ had the best consistency with experimental distribution function shown in different diagrams for each season. Results also showed that the mean wave energy in the research period was about 49.88 k W/m and the maximum density of wave energy was found in February and March, 2010.

Mapping and Functional Analysis of LE Gene in a Lethal Etiolated Rice Mutant at Seedling Stage

An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorophyll b,total chlorophyll,and carotenoids.Additionally,the mutant displayed a significantly decreased number of chloroplast grana,along with irregular and less-stacked grana lamellae.The le mutant showed markedly diminished root length,root surface area,and root volume compared with the wild type.It also exhibited significantly lower catalase activity and total protein content,while peroxidase activity was significantly higher.Using the map-based cloning method,we successfully mapped the LE gene to a 48-kb interval between markers RM16107 and RM16110 on rice chromosome 3.A mutation(from T to C)was identified at nucleotide position 692 bp of LOC_Os03g59640(ChlD),resulting in a change from leucine to proline.By crossing HM133(a pale green mutant with a single-base substitution of A for G in exon 10 of ChlD subunit)with a heterozygous line of le(LEle),we obtained two plant lines heterozygous at both the LE and HM133 loci.Among 15 transgenic plants,3 complementation lines displayed normal leaf color with significantly higher total chlorophyll,chlorophyll a,and chlorophyll b contents.The mutation in le led to a lethal etiolated phenotype,which has not been observed in other ChlD mutants.The mutation in the AAA+domain of ChlD disrupted the interaction of ChlDle with ChlI as demonstrated by a yeast two-hybrid assay,leading to the loss of ChlD function and hindering chlorophyll synthesis and chloroplast development.Consequently,this disruption is responsible for the lethal etiolated phenotype in the mutant.

Cloning of PsMYB62 and analysis of cadmium resistant in Potentilla sericea

Potentilla sericea is a heavy metal hyperaccumulator landscaping plant.MYB transcription factors play an important role in regulating plant stress response to adversity.However,there are few studies on MYB transcription factors in stress tolerance in Potentilla sericea.In this study,the PsMYB62 gene was successfully cloned from Potentilla sericea.Methods:Bioinformatic analysis and real-time quantitative PCR(qPCR)methods were used to evaluate this gene.The transgenic A.thaliana were obtained by flower dipping and the gene function was identified by determining physiological indicators under cadmium stress.Results:The open reading frame of PsMYB62 is 942 bp,which encodes 313 amino acids(aa)and belongs to the R2R3 MYB transcription factor.The plant overexpression vector PBI121-PsMYB62-GFP was constructed and successfully transferred into A.thaliana.The relative expression level of PsMYB62 was significantly increased by CdCl_(2),NaCl,ABA,and mannitol treatments.The germination rate of transgenic seeds was higher than those of wild type(WT)and empty vector(EV)under different concentrations of cadmium treatment.Upon treatment with 100μmol·L^(−1)of CdCl_(2)·2.5H_(2)O,the activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT)in the transgenic plants were significantly higher than those in the WT and EV.The contents of H_(2)O_(2),O_(2)·−and malondialdehyde(MDA)in transgenic lines were increased,but lower than those in WT and EV.The expression levels of AtGSH,AtPCS,and AtNAS4 that were related to the regulation of cadmium were increased,but the expression levels of transgenic lines were higher than those of WT and EV.Conclusion:The above results showed that PsMYB62 could be induced by cadmium and could improve the cadmium resistance of plants.

Cloning and Analysis of Genes SAMS From Glycine soja

S-adenosylmethionine （SAM） plays important role in trans-methyl reactions. Under the condition of drought （30% PEG）, salinity （200 mmol· L^-1 NaCl） and low temperature （4℃）, total RNA was extracted from the leaf and the first strand of cDNA was synthesized with reverse transcription. S-adenosylmethionine synthetase gene （SAMS gene） was amplified by PCR with the first strand cDNA as template and a pair of primers which was based on constructed ESTs sequence. Full-length SAMS gene sequence was obtained by BLAST comparison. According to the analysis, completed sequence of SAMS gene was integrality. The sequence of the SAMS gene was 1 185 bp in length with an opening reading frame （ORF） encoding 394 amino acids. The cDNA sequence showed a significant homology to the SAM genes from Phaseolus lunatus （89%）, Medicago sativa （85%）. A prokaryotic expression vectors based on pET-32b had been constructed and prokaryotic expression was analyzed in order to lay a strong foundation for resist adversity function analysis through situation of genic expression analysis.

Industrial Food Quality Analysis Using New k-Nearest-Neighbour methods

The problem of predicting continuous scalar outcomes from functional predictors has received high levels of interest in recent years in many fields,especially in the food industry.The k-nearest neighbor(k-NN)method of Near-Infrared Reflectance(NIR)analysis is practical,relatively easy to implement,and becoming one of the most popular methods for conducting food quality based on NIR data.The k-NN is often named k nearest neighbor classifier when it is used for classifying categorical variables,while it is called k-nearest neighbor regression when it is applied for predicting noncategorical variables.The objective of this paper is to use the functional Near-Infrared Reflectance(NIR)spectroscopy approach to predict some chemical components with some modern statistical models based on the kernel and k-Nearest Neighbour procedures.In this paper,three NIR spectroscopy datasets are used as examples,namely Cookie dough,sugar,and tecator data.Specifically,we propose three models for this kind of data which are Functional Nonparametric Regression,Functional Robust Regression,and Functional Relative Error Regression,with both kernel and k-NN approaches to compare between them.The experimental result shows the higher efficiency of k-NN predictor over the kernel predictor.The predictive power of the k-NN method was compared with that of the kernel method,and several real data sets were used to determine the predictive power of both methods.

Association between chromosomal aberration of COX8C and tethered spinal cord syndrome:array-based comparative genomic hybridization analysis

Copy number variations have been found in patients with neural tube abnormalities.In this study,we performed genome-wide screening using high-resolution array-based comparative genomic hybridization in three children with tethered spinal cord syndrome and two healthy parents.Of eight copy number variations,four were non-polymorphic.These non-polymorphic copy number variations were associated with Angelman and Prader-Willi syndromes,and microcephaly.Gene function enrichment analysis revealed that COX8 C,a gene associated with metabolic disorders of the nervous system,was located in the copy number variation region of Patient 1.Our results indicate that array-based comparative genomic hybridization can be used to diagnose tethered spinal cord syndrome.Our results may help determine the pathogenesis of tethered spinal cord syndrome and prevent occurrence of this disease.

Developing Reliable Digital Healthcare Service Using Semi-Quantitative Functional Resonance Analysis

Since entering the era of Industry 4.0,the concept of Healthcare 4.0 has also been put forward and explored by researchers.How to use Information Technology(IT)to better serve people’s healthcare is one of the most featured emerging directions in the academic circle.An important field of Healthcare 4.0 research is the reliability engineering of healthcare service.Because healthcare systems often affect the health and even life of their users,developers must be very cautious in the design,development,and operation of these healthcare systems and services.The problems to be solved include the reliability of business process,system functions,and personal healthcare data.The Functional Resonance Analysis Method(FRAM)has been applied in reliability engineering for safety-critical systems in available studies,using both qualitative and quantitative approaches.However,the method has not been applied in the field of digital healthcare services development.Therefore,to narrow the gap,we present in this paper a semi-quantitative functional resonance analysis method to develop reliable healthcare services for diabetics.Moreover,this paper has tried to improve the reliability design of the service-oriented architecture(SOA)of traditional insulin pump therapy by system thinking.

Function Words Analysis——A Reading Comprehension Aid for Chinese Engineers

Background Generally speaking. Chinese college graduates in the fifties and sixties took Russian as their second language, and those who graduated in the seventies had no second language to speak of. Now, in the years of our Open Door Policy, they find they have to learn some English and learn it quickly. They try to learn from radio and TV and many take English courses of 4 to 6 months, with varying degree of success. Their chief stumbling blocks

Functional Discourse Analysis of Andersen＇s ＂The Little Match-Seller＂

Within a framework of Halliday＇s SFL （Systemic Functional Linguistics）, this paper takes a functional approach to the analysis of the fairy tale--＂The Little Match-Seller＂ （Andersen, 1846）. The analysis is carried out in three respects： genre, register and language. Genre analysis indicates that the schematic structure of the fairy tale is very much in line with the one mentioned by Hasan. Register analysis shows that the material process is the most dominant one; the Finite in the clauses is predominantly in past tenses, and most of the Themes in the text are unmarked ones. Language analysis reveals that the lexical density in the fairy tale is relatively low. The findings in this paper can help deepen people＇s understanding of the textual features of English fairy tales.

Cloning and expression analysis of HXK1 gene from Jatropha curcas L.

HXK(Hexokinase)gene family and the role of JcHXK1 in Jatropha curcas L.were explored.Totally 4 HXK genes JcHXK1,JcHXK2,JcHXK3 and JcHKL1 were identified and complete ORF of JcHXK1 was cloned.Functional domain,phylogenetic evolution and low-temperature expression characteristics were analyzed.Results showed that full-length JcHXK1 cDNA was 1497 bp,encoding 498 amino acids with molecular weight of 53.81 kDa and pI of 5.03.Further phylogenetic evolutionary analysis demonstrated HXK1 protein was clustered into 6 small branches and 2 large branches.Sequence alignment showed that HXK1 contained several conserved glycine residues and hydrophobic channels.Prokaryotic expression vector of JcHXK1 was constructed and 12%SDS-PAGE detection showed that it was highly expressed in E.coli.These research was expected to lay a foundation for further gene functional verification and cold signal transduction mechanism for HXK1 in Jatropha curcas L.

Transcriptome and Functional Analysis of Fiber-related Gene Expression in Cotton

Fiber cell initiation is a complex process involving many pathways,including phytohormones and components for transcriptional and posttranscriptional regulation.Here we report expression