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.

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.

Abnormal function of EPHA2/p.R957P mutant in congenital cataract

AIM:To identify genetic defects in a Chinese family with congenital posterior polar cataracts and assess the pathogenicity.METHODS:A four-generation Chinese family affected with autosomal dominant congenital cataract was recruited.Nineteen individuals took part in this study including 5 affected and 14 unaffected individuals.Sanger sequencing targeted hot-spot regions of 27 congenital cataract-causing genes for variant discovery.The pathogenicity of the variant was evaluated by the guidelines of American College of Medical Genetics and InterVar software.Confocal microscopy was applied to detect the subcellular localization of fluorescence-labeled ephrin type-A receptor 2(EPHA2).Co-immunoprecipitation assay was implemented to estimate the interaction between EphA2 and other lens membrane proteins.The mRNA and protein expression were analyzed by reverse transcription-polymerase chain reaction(qRT-PCR)and Western blotting assay,respectively.The cell migration was analyzed by wound healing assay.Zebrafish model was generated by ectopic expression of human EPHA2/p.R957P mutant to demonstrate whether the mutant could cause lens opacity in vivo.RESULTS:A novel missense and pathogenic variant c.2870G>C was identified in the sterile alpha motif(SAM)domain of EPHA2.Functional studies demonstrated the variant’s impact:reduced EPHA2 protein expression,altered subcellular localization,and disrupted interactions with other lens membrane proteins.This mutant notably enhanced human lens epithelial cell migration,and induced a central cloudy region and roughness in zebrafish lenses with ectopic expression of human EPHA2/p.R957P mutant under differential interference contrast(DIC)optics.CONCLUSION:Novel pathogenic c.2870G>C variant of EPHA2 in a Chinese congenital cataract family contributes to disease pathogenesis.

Function Analysis of Urban Rail Transit Smart Station

In order to make rail transit smart stations serve passengers better,the potential of smart stations should be optimized to reduce time cost,and relieve traffic congestion.In this paper,the construction of smart stations based on the management experience of a subway station and the benefits of smart stations were analyzed and discussed.The construction of smart stations,as well as the key technologies for the construction of smart stations,will aid in the automation and intelligent management of subway stations.

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.

Outlier Detection of Air Quality for Two Indian Urban Cities Using Functional Data Analysis

Human living would be impossible without air quality. Consistent advancements in practically every aspect of contemporary human life have harmed air quality. Everyday industrial, transportation, and home activities turn up dangerous contaminants in our surroundings. This study investigated two years’ worth of air quality and outlier detection data from two Indian cities. Studies on air pollution have used numerous types of methodologies, with various gases being seen as a vector whose components include gas concentration values for each observation per-formed. We use curves to represent the monthly average of daily gas emissions in our technique. The approach, which is based on functional depth, was used to find outliers in the city of Delhi and Kolkata’s gas emissions, and the outcomes were compared to those from the traditional method. In the evaluation and comparison of these models’ performances, the functional approach model studied well.

Classification of Service Functions in the Life Circle of Beijing Subway Line 7

As a key node for the surrounding area,metro stations are closely connected with the surrounding urban space and develop cooperatively.Different types of metro stations have differences in land use and functional positioning.This paper mainly used the methods of Tyson polygon,kernel density analysis and correlation analysis,based on POI data,to classify the stations of Beijing Metro Line 7.This paper made a detailed analysis of commercial subway stations,and analyzed the distribution characteristics of commercial metro stations on Line 7.

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.

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.

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.

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.

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.

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

A Functional Stylistic Analysis of“Do not Go Gentle into That Good Night”

As two main approaches to analyzing literary discourses,literary criticism and stylistic analysis have been employed for a long period of time.With the application of the theories of Functional Grammar,this paper is written for the purpose of being helpful for the readers of Dylan Thomas’poem“Do not Go Gentle into That Good Night”to understand the poem’s theme better as well as appreciate the artistic adaptation of language.Furthermore,because the poem is analyzed with the application of Functional Grammar,it helps the readers to appreciate“Do not Go Gentle into That Good Night”from a new perspective.

Analysis of the Longitudinal Structure Function F_L from the Non-linear Regge Gluon Density Behavior at Low-x

The longitudinal structure function with shadowing correction according to the nonlinear effects of the gluon density behavior at low x is considered. The solution of the GLR-MQ evolution equation for the gluon density shows that the FL^g（x, Q2） behavior can be tamed by the singularity at low x values. Comparing our results with H1 data at R=4 GeV-1 shows that at very low x this behavior is completely tamed by taking shadowing correction into account.

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.

Three-dimensional Analysis of Functionally Graded Piezoelectric Plate with Arbitrarily Distributed Material Properties

An orthotropic functionally graded piezoelectric rectangular plate with arbitrarily distributed material properties was studied, which is simply supported and grounded（electrically） on its four lateral edges. The state equations of the functionally graded piezoelectric material were obtained using the state-space approach, and a Peano-Baker series solution was obtained for the coupled electroelastic fi elds of the functionally graded piezoelectric plate subjected to mechanical and electric loading on its upper and lower surfaces. The influence of different distributions of material properties on the structural response of the plate was studied using the obtained solutions.

INTEGRATION OF A ZONAL FUNCTION RELATED TO THE LOCAL HARMONIC ANALYSIS ON SPHERES

In studying Local harmonic analysis on the sphere S',R.S.Strichartz introduced certain zonab functionsid(x,y))which satisfy the equation