Black wings of butterfly Ornithoptera goliath and infrared-band radiative cooling function of Rapala dioetas butterfly wings are associated with black pigment(e.g.,melanin)and unique hierarchical micro/nanostructures,...Black wings of butterfly Ornithoptera goliath and infrared-band radiative cooling function of Rapala dioetas butterfly wings are associated with black pigment(e.g.,melanin)and unique hierarchical micro/nanostructures,greatly stimulating biomimetic fabrication of functional photonic structures but mainly targeted to one prototype.Targeted at two-prototype integrated biomimetic fabrication from fully compositional/structural/functional aspects,femtosecond(fs)laser subtractive/additive-integrated hierarchical micro/nano-manufacturing technique is proposed in this work.This technique can one-step transfer refractory metals(e.g.,W,Mo,Nb,Ta)into black non-stoichiometric oxide nanomaterials with abundant oxygen vacancies and simultaneously enable the realization of in situ quasi-controllable micro/nanoscale hierarchical aggregation and assembly,all displaying black color but with tunable infrared emission.Adjusting the scan interval for biomimetic manufacturing can tailor the structural oxidation degree,the emission in the long-wave infrared(LWIR)band while keeping the blackness of hierarchical aggregates,and the confined height between the covering quartz plate and the ablated sample.The blackening efficiency of this technique can reach∼11.25 cm^(2)·min^(−1),opening opportunities for high-throughput optical/thermal applications.Selectively patterned Chinese characters,Arabic numbers,and English letters are easily fabricable,which are intrinsically invisible-infrared dual-band encrypted but decryptable via static/dynamic environment stimuli(e.g.,sample heating/cooling,introducing external hot/cold sources including human hands).The self-evolution from‘orderless’structuring to‘ordered’functionalization is validated for the proposed fs laser subtractive/additive-integrated biomimetic manufacturing,specifically from the synthesis of diverse black nanomaterials and the seemingly disordered micro/nano-aggregates to the ordered optical/thermal regulation capacities for a delicate modulation of information encryption and decryption,unveiling a new concept for future exploration and extension.展开更多
To investigate the expression profile of maize genes induced by submergence, a subtracted cDNA library of maize seedling roots was constructed using suppression subtractive hybridization (SSH). The cDNA of maize seedl...To investigate the expression profile of maize genes induced by submergence, a subtracted cDNA library of maize seedling roots was constructed using suppression subtractive hybridization (SSH). The cDNA of maize seedling roots treated with submergence (ST) was used as tester and what from untreated roots (UT) as driver. Products of the secondary PCR from the forward subtraction were cloned into T/A vector and transferred into Escherichia coli strain JM10B by electroporation. Four hundred and eight randomly chosen transformants carrying cDNA fragments were screened with PCR-Select Deferential Screening Kit. One hundred and eighty-four cDNA clones were identified as, submergence specifically induced or highly expressed. After sequencing and removing redundant cDNAs, we got 95 submergence-induced cDNA clones. Of the 95 cDNA clones, 68 contain the regions with 60%-90% identity to their homolog in GenBank, 21 are expected to be novel genes, only 6 correspond to the published maize sequences.展开更多
In order to improve the accuracy and reliability of the driving fatigue detection based on a single feature, a new detection algorithm based on multiple features is proposed. Two direct driver＇s facial features refle...In order to improve the accuracy and reliability of the driving fatigue detection based on a single feature, a new detection algorithm based on multiple features is proposed. Two direct driver＇s facial features reflecting fatigue and one indirect vehicle behavior feature indicating fatigue are considered. Meanwhile, T-S fuzzy neural network（TSFNN）is adopted to recognize the driving fatigue of drivers. For the structure identification of the TSFNN, subtractive clustering（SC） is used to confirm the fuzzy rules and their correlative parameters. Moreover, the particle swarm optimization （PSO）algorithm is improved to train the TSFNN. Simulation results and experiments on vehicles show that the proposed algorithm can effectively improve the convergence speed and the recognition accuracy of the TSFNN, as well as enhance the correct rate of driving fatigue detection.展开更多
Phosphorus is one of the three essential macroelements for plant growth. Plants respond to phosphorus starvation through adaptive mechanisms involved in morphological, biochemical and molecular changes. To investigate...Phosphorus is one of the three essential macroelements for plant growth. Plants respond to phosphorus starvation through adaptive mechanisms involved in morphological, biochemical and molecular changes. To investigate the molecular background of the adaptive mechanisms, the suppression subtractive hybridization (SSH) method was used to construct a rice phosphorus-starvation ( Pi-starvation) induced cDNA library. Through screening of the cDNA library and sequencing of the enriched cDNAs, 18 known genes and 47 novel genes were identified. The known genes are involved in different metabolic processes, including phosphate uptake and transport, signal transduction, protein synthesis and degradation, carbon metabolism and stress response. Northern analysis was performed to detect the expression patterns of some known genes and novel genes under different phosphorus levels. Different expression patterns of the selected genes were identified, which suggests that genes involved in different pathways may have different responses to Pi-starvation.展开更多
This paper presents a multi-face detection method for color images. The method is based on the assumption that faces are well separated from the background by skin color detection. These faces can be located by the pr...This paper presents a multi-face detection method for color images. The method is based on the assumption that faces are well separated from the background by skin color detection. These faces can be located by the proposed method which modifies the subtractive clustering. The modified clustering algorithm proposes a new definition of distance for multi-face detection, and its key parameters can be predetermined adaptively by statistical information of face objects in the image. Downsampling is employed to reduce the computation of clustering and speed up the process of the proposed method. The effectiveness of the proposed method is illustrated by three experiments.展开更多
New materials and manufacturing technologies require applicable non-destructive techniques for quality assurance so as to achieve better performance.This study comprehensively investigated the effect of influencing fa...New materials and manufacturing technologies require applicable non-destructive techniques for quality assurance so as to achieve better performance.This study comprehensively investigated the effect of influencing factors includ-ing excitation frequency,lift-off distance,defect depth and size,residual heat,and surface roughness on the defect EC signals of an Inconel 738LC alloy produced by selective laser melting(SLM).The experimental investigations recorded the impedance amplitude and phase angle of EC signals for each defect to explore the feasibility of detecting sub-surface defects by merely analyzing these two key indicators.Overall,this study revealed preliminary qualitative and roughly quantitative relationships between influencing factors and corresponding EC signals,which provided a prac-tical reference on how to quantitively inspect subsurface defects using eddy current testing(ECT)on SLMed parts,and also made solid progress toward on-line ECT in additive/subtractive hybrid manufacturing(ASHM)for fabricating SLMed parts with enhanced quality and better performance.展开更多
In this study, a forward cDNA library was constructed by suppression subtractive hybridization using seedling leaves of CN165, a drought-tolerant maize inbred line. In the suppression subtractive hybridization （SSH）...In this study, a forward cDNA library was constructed by suppression subtractive hybridization using seedling leaves of CN165, a drought-tolerant maize inbred line. In the suppression subtractive hybridization （SSH） library, 672 positive clones were picked up randomly. After polymerase chain reaction （PCR） of each clone, all the single clones were sequenced. Totally 598 available sequences were obtained. After cluster analysis of the EST sequences, 80 uniESTs were obtained, among which 57 uniESTs were contigs and 23 uniESTs were singlets. The results of BLASTN showed that all the uniESTs had homologous sequences in the nr database. The BLASTX results indicated that 68 uniESTs had significant protein homology, 8 uniESTs with homology of unknown proteins and putative proteins, and 4 uniESTs without protein homology. Those drought stress-induced genes were involved in many metabolism pathways to regulate plant growth and development under drought stress.展开更多
AIM: To identify potential diagnostic target genes in early reperfusion periods following warm liver ischemia before irreversible liver damage occurs. METHODS: We used two strategies (SSH suppression subtractive hybri...AIM: To identify potential diagnostic target genes in early reperfusion periods following warm liver ischemia before irreversible liver damage occurs. METHODS: We used two strategies (SSH suppression subtractive hybridization and hybridization of cDNA arrays) to determine early changes in gene expression profiles in a rat model of partial WI/R, comparing postischemic and adjacent nonischemic liver lobes. Differential gene expression was verified (WI/R; 1 h/2 h) and analyzed in more detail after warm ischemia (1 h) in a reperfusion time kinetics (0, 1, 2 and 6 h) and compared to untreated livers by Northern blot hybridizations. Protein expression was examined on Western blots and by immunohistochemistry for four differentially expressed target genes (Hsp70, Hsp27, Gadd45a and IL-1rI). RESULTS: Thirty-two individual WI/R target genes showing altered RNA levels after confirmation by Northern blot analyzes were identified. Among them, six functionally uncharacteristic expressed sequences and 26 known genes (12 induced in postischemic liver lobes, 14 with higher transcriptional expression in adjacent nonischemic liver lobes). Functional categories of the verified marker genes indicate on the one hand cellular stress and tissue damage but otherwise activation of protective cellular reactions (AP-1 transcription factors, apoptosis related genes, heat shock genes). In order to assign the transcriptional status to the biological relevant protein level we demonstrated that Hsp70, Hsp27, Gadd45a and IL-1rI were clearly up-regulated comparing postischemic and untreated rat livers, suggesting their involvement in the WI/R context. CONCLUSION: This study unveils a WI/R response gene set that will help to explore molecular pathways involved in the tissue damage after WI/R. In addition, these genes especially Hsp70 and Gadd45a might represent promising new candidates indicating WI/R liver damage.展开更多
In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After ...In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.展开更多
AIM： To screen for metronidazole （MTZ）-resistance associated gene fragments of H pylori by suppression subtractive hybridization （SSH）. METHODS： Five MTZ-resistant （tester, T） and 1 MTZ- susceptible （driver, ...AIM： To screen for metronidazole （MTZ）-resistance associated gene fragments of H pylori by suppression subtractive hybridization （SSH）. METHODS： Five MTZ-resistant （tester, T） and 1 MTZ- susceptible （driver, D） clinical H pylori isolates were selected. Genomic DNAs were prepared and submitted to Rsa I digestion. Then two different adaptors were ligated respectively to the 5＇-end of two aliquots of the tester DNA fragments and SSH was made between the tester and driver DNAs. The specific inserts of tester strains were screened and MTZ-resistance related gene fragments were identified by dot blotting. RESULTS： Among the randomly selected 120 subtractive colonies, 37 DNA fragments had a different number of DNA copies （≥ 2 times） in resistant and susceptible strains and 17 of them had a significantly different number of DNA copies （≥ 3 times）. Among the sequences obtained from the 17 DNA fragments, new sequences were found in 10 DNA fragments and duplicated sequences in 7 DNA fragments, representing respectively the sequences of depeptide ABC transporter periplasmic dipeptide-binding protein （dppA）, permease protein （dppB）, ribosomal protein S4 （rps4）, ribonuclease Ⅲ （rnc）, protease （pqqE）, diaminopimelate epimerase （dapF）, acetatekinase （ackA）, H pylori plasmid pHP51 and Hpylori gene 1334. CONCLUSION： Gene fragments specific to MTZ-resistant H pylori strains can be screened by SSH and may be associated with MTZ-resistant Hpylori.展开更多
A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examin...A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examined by differential screening. Of these, 185 clones were selected to be sequenced. A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database. Semi-quantitative RT-PCR was used to confirm further the expression profiles for 12 transcripts identified within the subtractive cDNA library. Gene ontology analyses indicated that many of the different genes identified have unknown or hypothetical functions while it is speculated that other genes play different mo- lecular roles. In our study, genes involved in bud dormancy release were growth-related or stress-responsive, while low-temperature-induced ribosomal proteins may also play a role in bud dormancy release. Our results provide interesting information for further understanding of the molecular mechanism of bud dormancy release in tree peonies.展开更多
Microbial population and enzyme activities are the significant indicators of soil strength.Soil microbial dynamics characterize microbial population and enzyme activities.The present study explores the development of ...Microbial population and enzyme activities are the significant indicators of soil strength.Soil microbial dynamics characterize microbial population and enzyme activities.The present study explores the development of efficient predictive modeling systems for the estimation of specific soil microbial dynamics,like rock phosphate solubilization,bacterial population,and ACC-deaminase activity.More specifically,optimized subtractive clustering(SC)and Wang and Mendel's(WM)fuzzy inference systems(FIS)have been implemented with the objective to achieve the best estimation accuracy of microbial dynamics.Experimental measurements were performed using controlled pot experiment using minimal salt media with rock phosphate as sole carbon source inoculated with phosphate solubilizing microorganism in order to estimate rock phosphate solubilization potential of selected strains.Three experimental parameters,including temperature,pH,and incubation period have been used as inputs SC-FIS and WM-FIS.The better performance of the SC-FIS has been observed as compared to the WM-FIS in the estimation of phosphate solubilization and bacterial population with the maximum value of the coefficient of determination(0.9988)2 R=in the estimation of previous microbial dynamics.展开更多
For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of Rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and f...For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of Rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and fed male ticks were constructed using suppression subtractive hybridization （SSH）. A total of 247 female expression sequence tags （ESTs） and 168 male ESTs were obtained from the two SSH cDNA libraries. It is predicted that 25 female ESTs and 44 female ESTs contain the 5＂ and 3＂ ends, respectively, and that 53 male ESTs and 74 male ESTs contain the 5＂ and 3＂ ends, respectively. To identify the subtraction rate of the two SSH cDNA libraries, the RT-PCR method was used to test 24 female ESTs and 21 male ESTs selected randomly but not repeatedly. The results showed that there were 13 upregulated or differentially expressed genes in the partially fed salivary gland of the female R. haemaphysaloides and that the differentially expressed rate was 54%. In addition, they indicated that there were 9 upregulated or differently expressed genes in the fed salivary gland of the male R. haemaphysaloides and that the differentially expressed rate was 43%. Putative translations of 141 （57%） female ESTs and 125 （74%） male ESTs had similarity to GenBank sequences, and 32 （23%） female ESTs and 29 （23%） male ESTs exhibited similarity to tick proteins, which showed that most of the proteins in the libraries were mainly related to the feeding blood physiology of the ticks.展开更多
In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After ...In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.展开更多
Soybean is planted worldwide and its productivity is significantly hampered by salinity. Development of salt tolerant cultivars is necessary for promoting soybean production. Despite wealth of information generated on...Soybean is planted worldwide and its productivity is significantly hampered by salinity. Development of salt tolerant cultivars is necessary for promoting soybean production. Despite wealth of information generated on salt tolerance mechanism, its basics still remain elusive. A continued effort is needed to understand the salt tolerance mechanism in soybean using suitable molecular tools. To better understand the molecular basis of the responses of soybean to salt stress and to get an enrichment of critical salt stress responsive genes in soybean, suppression subtractive hybridization libraries （SSH） are constructed for the root tissue of two cultivated soybean genotypes, one was tolerant and the other was sensitive to salt stress. To compare the responses of plants in salt treatment and non-treatment, SSH1 was constructed for the salt-tolerant cultivar Wenfeng 7 and SSH2 was constructed for the salt-sensitive cultivar Union. From the two SSH cDNA libraries, a total of 379 high quality ESTs were obtained. These ESTs were then annotated by performing sequence similarity searches against the NCBI nr （National Center for Biotechnology Information protein non-redundant） database using the BLASTX program. Sixty-three genes from SSH1 and 49 genes from SSH2 could be assigned putative function. On the other hand, 25 ESTs of SSH1 which may be not the salt tolerance-related genes were removed by comparing and analyzing the ESTs from the two S SH libraries, which increased the proportion of the genes related to salt tolerance in S SH 1. These results suggested that the novel way could realize low background of SSH and high level enrichment of target cDNAs to some extent.展开更多
Additive manufacturing(AM)processes are reliable techniques to build highly complex metallic parts.Direct energy deposition(DED)is one of the most common technologies to 3D print metal alloys.Despite a wide range of l...Additive manufacturing(AM)processes are reliable techniques to build highly complex metallic parts.Direct energy deposition(DED)is one of the most common technologies to 3D print metal alloys.Despite a wide range of literature that has discussed the ability of DED in metal printing,weak binding,poor accuracy,and rough surface still exist in final products.Thus,limitations in 3D printing of metal powder and wire indicate post-processing techniques required to achieve high quality in both mechanical properties and surface quality.Therefore,hybrid manufacturing(HM),specifically additive/subtractive hybrid manufacturing(ASHM)of DED has been proposed to enhance product quality.ASHM is a capable process that combines two technologies with 3-axis or multi-axis machines.Different methods have been suggested to increase the accuracy of machines to find better quality and microstructure.In contrast,drawbacks in ASHM still exist such as limitations in existing reliable materials and poor accuracy in machine coordination to avoid collision in the multi-axes machine.It should be noted that there is no review work with focuses on both DED and hybridization of DED processes.Thus,in this review work,a unique study of DED in comparison to ASHM as well as novel techniques are discussed with the objective of showing the capabilities of each process and the benefits of using them for different applications.Finally,new gaps are discussed in ASHM to enhance the layer bonding and surface quality with the processes'effects on microstructures and performance.展开更多
A novel approach for constructing robust Mamdani fuzzy system was proposed, which consisted of an efficiency robust estimator(partial robust M-regression, PRM) in the parameter learning phase of the initial fuzzy syst...A novel approach for constructing robust Mamdani fuzzy system was proposed, which consisted of an efficiency robust estimator(partial robust M-regression, PRM) in the parameter learning phase of the initial fuzzy system, and an improved subtractive clustering algorithm in the fuzzy-rule-selecting phase. The weights obtained in PRM, which gives protection against noise and outliers, were incorporated into the potential measure of the subtractive cluster algorithm to enhance the robustness of the fuzzy rule cluster process, and a compact Mamdani-type fuzzy system was established after the parameters in the consequent parts of rules were re-estimated by partial least squares(PLS). The main characteristics of the new approach were its simplicity and ability to construct fuzzy system fast and robustly. Simulation and experiment results show that the proposed approach can achieve satisfactory results in various kinds of data domains with noise and outliers. Compared with D-SVD and ARRBFN, the proposed approach yields much fewer rules and less RMSE values.展开更多
In order to isolate genes related to anther development and understand molecular basis of male sterility,cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization（SSH） approach.672 d...In order to isolate genes related to anther development and understand molecular basis of male sterility,cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization（SSH） approach.672 different expressed clones were selected from the fertile disk floret buds.PCR results showed that cDNA inserts were ranged from 100 to 750 bp.303 positive clones screened by dot-blot hybridization were sequenced.273 out of 303 sequenced clones produced readable sequences;these sequences represent 87 non-repetitive sequences.The homology alignment showed that 76 expressed sequence tags（ESTs） had functional annotations in GenBank,the other 11 ESTs without any homology to the known gene.In addition,87 ESTs were divided into 17 groups according to MIPS of Arabidopsis thaliana database.Sequence data from the cDNA library have been deposited with the GenBank under the accession numbers GT067016-GT067085.As an important result in this study,7 genes related to anther development were isolated.Results from semi-quantitative RT-PCR showed 6 genes were expressed only in disk florets of fertile plants compared with that of male sterile plants.These ESTs obtained provide important clues for further isolation and identification of fertility-related genes in Z.elegans.展开更多
Monitoring expression at the transcriptional level is the first essential step for the functional analysis of plant genes. Genes-encoding proteins directly involved in early response to elicitor constitute only a smal...Monitoring expression at the transcriptional level is the first essential step for the functional analysis of plant genes. Genes-encoding proteins directly involved in early response to elicitor constitute only a small fraction of all the genes affected by elicitor. Transcriptional responses to various elicitors have been extensively studied in different plants including Nicotiana and Arabidopsis thaliana; however, corresponding data aren＇t available for non-heading Chinese cabbage. To address this problem, we describe a suppression subtractive library-based approach to isolate the plant＇s ESTs up-regulated in the early induction/execution of the HR induced by elicitor PB90 from Phytophthora boehmeriae. According to their putative identification in BLAST searches against the three genome databases, 70 up-regulated genes were classified into 9 parts： some aspect of primary ＇metabolism＇ or ＇energy＇ production; ＇protein synthesis＇ or ＇protein fate＇; cellular communication/signal transduction mechanism; cell fates including Beclin, SPT1, and SPT2; HLA-B and AGO1 which participate in transcription; cellular transport and hypothetical proteins or proteins for which a function has yet to be determined. Seven selected genes such as Beclin, thioredoxin, HLA-B, MAP3K, SPT1, SPT2, and AGO1 were up-regulated induced by PB90, suggesting that the genes may play an important role in PB90-triggered HR.展开更多
AIM： To clone and identify human genes transactivated by PSITP5 by constructing a cDNA subtractive library with suppression subtractive hybridization （SSH） technique. METHODS： SSH and bioinformatics techniques wer...AIM： To clone and identify human genes transactivated by PSITP5 by constructing a cDNA subtractive library with suppression subtractive hybridization （SSH） technique. METHODS： SSH and bioinformatics techniques were used for screening and cloning of the target genes transactivated by PS1TP5 protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1（-）- myc-his（A）-PS1TP5 and pcDNA3.1（-）-myc-his（A） empty vector, respectively, and SSH technique was employed to analyze the differentially expressed DNA sequence between the two groups. After digestion with restriction enzyme Rsa Ⅰ, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. The tester cDNA was hybridized with driver cDNA twice and subjected to nested PCR for two times, and then subcloned into T/A plasmid vectors to set up the subb-active library. Amplification of the library was carried out with E.. coil strain DH5α. The cDNA was sequenced and analyzed in GenBank with Vector NTI 9.1 and NCBI BLAST software after PCR amplification. RESULTS： The subtractive library of genes transactivated by PS1TP5 was constructed successfully. The amplified library contained 90 positive clones. Colony PCR showed that 70 clones contained 200-1000-bp inserts. Sequence analysis was performed in 30 clones randomly, and the full-length sequences were obtained by bioinformatics technique. Altogether 24 coding sequences were obtained, which consisted of 23 known and 1 unknown.One novel gene with unknown functions was found and named as PSITP5TP1 after being electronically spliced, and deposited in GenBank （accession number： DQ487761）. CONCLUSION： PSITP5 is closely correlated with immunoregulation, carbohydrate metabolism, signal transduction, formation mechanism of hepatic fibrosis, and occurrence and development of tumor. Understanding PSlTP5 transactive proteins may help to bring some new clues for further studying the biological functions of pre-S1 protein.展开更多
基金financially supported by Shanghai Pujiang Program 23PJ1406500.
文摘Black wings of butterfly Ornithoptera goliath and infrared-band radiative cooling function of Rapala dioetas butterfly wings are associated with black pigment(e.g.,melanin)and unique hierarchical micro/nanostructures,greatly stimulating biomimetic fabrication of functional photonic structures but mainly targeted to one prototype.Targeted at two-prototype integrated biomimetic fabrication from fully compositional/structural/functional aspects,femtosecond(fs)laser subtractive/additive-integrated hierarchical micro/nano-manufacturing technique is proposed in this work.This technique can one-step transfer refractory metals(e.g.,W,Mo,Nb,Ta)into black non-stoichiometric oxide nanomaterials with abundant oxygen vacancies and simultaneously enable the realization of in situ quasi-controllable micro/nanoscale hierarchical aggregation and assembly,all displaying black color but with tunable infrared emission.Adjusting the scan interval for biomimetic manufacturing can tailor the structural oxidation degree,the emission in the long-wave infrared(LWIR)band while keeping the blackness of hierarchical aggregates,and the confined height between the covering quartz plate and the ablated sample.The blackening efficiency of this technique can reach∼11.25 cm^(2)·min^(−1),opening opportunities for high-throughput optical/thermal applications.Selectively patterned Chinese characters,Arabic numbers,and English letters are easily fabricable,which are intrinsically invisible-infrared dual-band encrypted but decryptable via static/dynamic environment stimuli(e.g.,sample heating/cooling,introducing external hot/cold sources including human hands).The self-evolution from‘orderless’structuring to‘ordered’functionalization is validated for the proposed fs laser subtractive/additive-integrated biomimetic manufacturing,specifically from the synthesis of diverse black nanomaterials and the seemingly disordered micro/nano-aggregates to the ordered optical/thermal regulation capacities for a delicate modulation of information encryption and decryption,unveiling a new concept for future exploration and extension.
文摘To investigate the expression profile of maize genes induced by submergence, a subtracted cDNA library of maize seedling roots was constructed using suppression subtractive hybridization (SSH). The cDNA of maize seedling roots treated with submergence (ST) was used as tester and what from untreated roots (UT) as driver. Products of the secondary PCR from the forward subtraction were cloned into T/A vector and transferred into Escherichia coli strain JM10B by electroporation. Four hundred and eight randomly chosen transformants carrying cDNA fragments were screened with PCR-Select Deferential Screening Kit. One hundred and eighty-four cDNA clones were identified as, submergence specifically induced or highly expressed. After sequencing and removing redundant cDNAs, we got 95 submergence-induced cDNA clones. Of the 95 cDNA clones, 68 contain the regions with 60%-90% identity to their homolog in GenBank, 21 are expected to be novel genes, only 6 correspond to the published maize sequences.
基金The National Key Technologies R & D Program during the 11th Five-Year Plan Period(No.2009BAG13A04)the Ph.D.Programs Foundation of Ministry of Education of China(No.200802861061)the Transportation Science Research Project of Jiangsu Province(No.08X09)
文摘In order to improve the accuracy and reliability of the driving fatigue detection based on a single feature, a new detection algorithm based on multiple features is proposed. Two direct driver＇s facial features reflecting fatigue and one indirect vehicle behavior feature indicating fatigue are considered. Meanwhile, T-S fuzzy neural network（TSFNN）is adopted to recognize the driving fatigue of drivers. For the structure identification of the TSFNN, subtractive clustering（SC） is used to confirm the fuzzy rules and their correlative parameters. Moreover, the particle swarm optimization （PSO）algorithm is improved to train the TSFNN. Simulation results and experiments on vehicles show that the proposed algorithm can effectively improve the convergence speed and the recognition accuracy of the TSFNN, as well as enhance the correct rate of driving fatigue detection.
文摘Phosphorus is one of the three essential macroelements for plant growth. Plants respond to phosphorus starvation through adaptive mechanisms involved in morphological, biochemical and molecular changes. To investigate the molecular background of the adaptive mechanisms, the suppression subtractive hybridization (SSH) method was used to construct a rice phosphorus-starvation ( Pi-starvation) induced cDNA library. Through screening of the cDNA library and sequencing of the enriched cDNAs, 18 known genes and 47 novel genes were identified. The known genes are involved in different metabolic processes, including phosphate uptake and transport, signal transduction, protein synthesis and degradation, carbon metabolism and stress response. Northern analysis was performed to detect the expression patterns of some known genes and novel genes under different phosphorus levels. Different expression patterns of the selected genes were identified, which suggests that genes involved in different pathways may have different responses to Pi-starvation.
文摘This paper presents a multi-face detection method for color images. The method is based on the assumption that faces are well separated from the background by skin color detection. These faces can be located by the proposed method which modifies the subtractive clustering. The modified clustering algorithm proposes a new definition of distance for multi-face detection, and its key parameters can be predetermined adaptively by statistical information of face objects in the image. Downsampling is employed to reduce the computation of clustering and speed up the process of the proposed method. The effectiveness of the proposed method is illustrated by three experiments.
基金Supported by Basic Research Project of Science and Technology Plan of Shenzhen(Grant No.JCYJ20170817111811303).
文摘New materials and manufacturing technologies require applicable non-destructive techniques for quality assurance so as to achieve better performance.This study comprehensively investigated the effect of influencing factors includ-ing excitation frequency,lift-off distance,defect depth and size,residual heat,and surface roughness on the defect EC signals of an Inconel 738LC alloy produced by selective laser melting(SLM).The experimental investigations recorded the impedance amplitude and phase angle of EC signals for each defect to explore the feasibility of detecting sub-surface defects by merely analyzing these two key indicators.Overall,this study revealed preliminary qualitative and roughly quantitative relationships between influencing factors and corresponding EC signals,which provided a prac-tical reference on how to quantitively inspect subsurface defects using eddy current testing(ECT)on SLMed parts,and also made solid progress toward on-line ECT in additive/subtractive hybrid manufacturing(ASHM)for fabricating SLMed parts with enhanced quality and better performance.
文摘In this study, a forward cDNA library was constructed by suppression subtractive hybridization using seedling leaves of CN165, a drought-tolerant maize inbred line. In the suppression subtractive hybridization （SSH） library, 672 positive clones were picked up randomly. After polymerase chain reaction （PCR） of each clone, all the single clones were sequenced. Totally 598 available sequences were obtained. After cluster analysis of the EST sequences, 80 uniESTs were obtained, among which 57 uniESTs were contigs and 23 uniESTs were singlets. The results of BLASTN showed that all the uniESTs had homologous sequences in the nr database. The BLASTX results indicated that 68 uniESTs had significant protein homology, 8 uniESTs with homology of unknown proteins and putative proteins, and 4 uniESTs without protein homology. Those drought stress-induced genes were involved in many metabolism pathways to regulate plant growth and development under drought stress.
文摘AIM: To identify potential diagnostic target genes in early reperfusion periods following warm liver ischemia before irreversible liver damage occurs. METHODS: We used two strategies (SSH suppression subtractive hybridization and hybridization of cDNA arrays) to determine early changes in gene expression profiles in a rat model of partial WI/R, comparing postischemic and adjacent nonischemic liver lobes. Differential gene expression was verified (WI/R; 1 h/2 h) and analyzed in more detail after warm ischemia (1 h) in a reperfusion time kinetics (0, 1, 2 and 6 h) and compared to untreated livers by Northern blot hybridizations. Protein expression was examined on Western blots and by immunohistochemistry for four differentially expressed target genes (Hsp70, Hsp27, Gadd45a and IL-1rI). RESULTS: Thirty-two individual WI/R target genes showing altered RNA levels after confirmation by Northern blot analyzes were identified. Among them, six functionally uncharacteristic expressed sequences and 26 known genes (12 induced in postischemic liver lobes, 14 with higher transcriptional expression in adjacent nonischemic liver lobes). Functional categories of the verified marker genes indicate on the one hand cellular stress and tissue damage but otherwise activation of protective cellular reactions (AP-1 transcription factors, apoptosis related genes, heat shock genes). In order to assign the transcriptional status to the biological relevant protein level we demonstrated that Hsp70, Hsp27, Gadd45a and IL-1rI were clearly up-regulated comparing postischemic and untreated rat livers, suggesting their involvement in the WI/R context. CONCLUSION: This study unveils a WI/R response gene set that will help to explore molecular pathways involved in the tissue damage after WI/R. In addition, these genes especially Hsp70 and Gadd45a might represent promising new candidates indicating WI/R liver damage.
基金This work was supported by Nationa1 NaturalScience Fundation of China No.39700148 and LifeScience Special fund of CAS supported by ChineseMinisery of Finance.
文摘In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.
基金Supported by the Natural Science Foundation of ZhejiangProvince, No. 29801
文摘AIM： To screen for metronidazole （MTZ）-resistance associated gene fragments of H pylori by suppression subtractive hybridization （SSH）. METHODS： Five MTZ-resistant （tester, T） and 1 MTZ- susceptible （driver, D） clinical H pylori isolates were selected. Genomic DNAs were prepared and submitted to Rsa I digestion. Then two different adaptors were ligated respectively to the 5＇-end of two aliquots of the tester DNA fragments and SSH was made between the tester and driver DNAs. The specific inserts of tester strains were screened and MTZ-resistance related gene fragments were identified by dot blotting. RESULTS： Among the randomly selected 120 subtractive colonies, 37 DNA fragments had a different number of DNA copies （≥ 2 times） in resistant and susceptible strains and 17 of them had a significantly different number of DNA copies （≥ 3 times）. Among the sequences obtained from the 17 DNA fragments, new sequences were found in 10 DNA fragments and duplicated sequences in 7 DNA fragments, representing respectively the sequences of depeptide ABC transporter periplasmic dipeptide-binding protein （dppA）, permease protein （dppB）, ribosomal protein S4 （rps4）, ribonuclease Ⅲ （rnc）, protease （pqqE）, diaminopimelate epimerase （dapF）, acetatekinase （ackA）, H pylori plasmid pHP51 and Hpylori gene 1334. CONCLUSION： Gene fragments specific to MTZ-resistant H pylori strains can be screened by SSH and may be associated with MTZ-resistant Hpylori.
基金supported by the Natural Science Foundation of Shangdong Province,China(Z2005D04).
文摘A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examined by differential screening. Of these, 185 clones were selected to be sequenced. A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database. Semi-quantitative RT-PCR was used to confirm further the expression profiles for 12 transcripts identified within the subtractive cDNA library. Gene ontology analyses indicated that many of the different genes identified have unknown or hypothetical functions while it is speculated that other genes play different mo- lecular roles. In our study, genes involved in bud dormancy release were growth-related or stress-responsive, while low-temperature-induced ribosomal proteins may also play a role in bud dormancy release. Our results provide interesting information for further understanding of the molecular mechanism of bud dormancy release in tree peonies.
文摘Microbial population and enzyme activities are the significant indicators of soil strength.Soil microbial dynamics characterize microbial population and enzyme activities.The present study explores the development of efficient predictive modeling systems for the estimation of specific soil microbial dynamics,like rock phosphate solubilization,bacterial population,and ACC-deaminase activity.More specifically,optimized subtractive clustering(SC)and Wang and Mendel's(WM)fuzzy inference systems(FIS)have been implemented with the objective to achieve the best estimation accuracy of microbial dynamics.Experimental measurements were performed using controlled pot experiment using minimal salt media with rock phosphate as sole carbon source inoculated with phosphate solubilizing microorganism in order to estimate rock phosphate solubilization potential of selected strains.Three experimental parameters,including temperature,pH,and incubation period have been used as inputs SC-FIS and WM-FIS.The better performance of the SC-FIS has been observed as compared to the WM-FIS in the estimation of phosphate solubilization and bacterial population with the maximum value of the coefficient of determination(0.9988)2 R=in the estimation of previous microbial dynamics.
基金the National Natural Science Foundation of China (31172095)
文摘For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of Rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and fed male ticks were constructed using suppression subtractive hybridization （SSH）. A total of 247 female expression sequence tags （ESTs） and 168 male ESTs were obtained from the two SSH cDNA libraries. It is predicted that 25 female ESTs and 44 female ESTs contain the 5＂ and 3＂ ends, respectively, and that 53 male ESTs and 74 male ESTs contain the 5＂ and 3＂ ends, respectively. To identify the subtraction rate of the two SSH cDNA libraries, the RT-PCR method was used to test 24 female ESTs and 21 male ESTs selected randomly but not repeatedly. The results showed that there were 13 upregulated or differentially expressed genes in the partially fed salivary gland of the female R. haemaphysaloides and that the differentially expressed rate was 54%. In addition, they indicated that there were 9 upregulated or differently expressed genes in the fed salivary gland of the male R. haemaphysaloides and that the differentially expressed rate was 43%. Putative translations of 141 （57%） female ESTs and 125 （74%） male ESTs had similarity to GenBank sequences, and 32 （23%） female ESTs and 29 （23%） male ESTs exhibited similarity to tick proteins, which showed that most of the proteins in the libraries were mainly related to the feeding blood physiology of the ticks.
文摘In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.
基金the National Natural Science Foundation of China (30771358)
文摘Soybean is planted worldwide and its productivity is significantly hampered by salinity. Development of salt tolerant cultivars is necessary for promoting soybean production. Despite wealth of information generated on salt tolerance mechanism, its basics still remain elusive. A continued effort is needed to understand the salt tolerance mechanism in soybean using suitable molecular tools. To better understand the molecular basis of the responses of soybean to salt stress and to get an enrichment of critical salt stress responsive genes in soybean, suppression subtractive hybridization libraries （SSH） are constructed for the root tissue of two cultivated soybean genotypes, one was tolerant and the other was sensitive to salt stress. To compare the responses of plants in salt treatment and non-treatment, SSH1 was constructed for the salt-tolerant cultivar Wenfeng 7 and SSH2 was constructed for the salt-sensitive cultivar Union. From the two SSH cDNA libraries, a total of 379 high quality ESTs were obtained. These ESTs were then annotated by performing sequence similarity searches against the NCBI nr （National Center for Biotechnology Information protein non-redundant） database using the BLASTX program. Sixty-three genes from SSH1 and 49 genes from SSH2 could be assigned putative function. On the other hand, 25 ESTs of SSH1 which may be not the salt tolerance-related genes were removed by comparing and analyzing the ESTs from the two S SH libraries, which increased the proportion of the genes related to salt tolerance in S SH 1. These results suggested that the novel way could realize low background of SSH and high level enrichment of target cDNAs to some extent.
文摘Additive manufacturing(AM)processes are reliable techniques to build highly complex metallic parts.Direct energy deposition(DED)is one of the most common technologies to 3D print metal alloys.Despite a wide range of literature that has discussed the ability of DED in metal printing,weak binding,poor accuracy,and rough surface still exist in final products.Thus,limitations in 3D printing of metal powder and wire indicate post-processing techniques required to achieve high quality in both mechanical properties and surface quality.Therefore,hybrid manufacturing(HM),specifically additive/subtractive hybrid manufacturing(ASHM)of DED has been proposed to enhance product quality.ASHM is a capable process that combines two technologies with 3-axis or multi-axis machines.Different methods have been suggested to increase the accuracy of machines to find better quality and microstructure.In contrast,drawbacks in ASHM still exist such as limitations in existing reliable materials and poor accuracy in machine coordination to avoid collision in the multi-axes machine.It should be noted that there is no review work with focuses on both DED and hybridization of DED processes.Thus,in this review work,a unique study of DED in comparison to ASHM as well as novel techniques are discussed with the objective of showing the capabilities of each process and the benefits of using them for different applications.Finally,new gaps are discussed in ASHM to enhance the layer bonding and surface quality with the processes'effects on microstructures and performance.
基金Project(61473298)supported by the National Natural Science Foundation of ChinaProject(2015QNA65)supported by Fundamental Research Funds for the Central Universities,China
文摘A novel approach for constructing robust Mamdani fuzzy system was proposed, which consisted of an efficiency robust estimator(partial robust M-regression, PRM) in the parameter learning phase of the initial fuzzy system, and an improved subtractive clustering algorithm in the fuzzy-rule-selecting phase. The weights obtained in PRM, which gives protection against noise and outliers, were incorporated into the potential measure of the subtractive cluster algorithm to enhance the robustness of the fuzzy rule cluster process, and a compact Mamdani-type fuzzy system was established after the parameters in the consequent parts of rules were re-estimated by partial least squares(PLS). The main characteristics of the new approach were its simplicity and ability to construct fuzzy system fast and robustly. Simulation and experiment results show that the proposed approach can achieve satisfactory results in various kinds of data domains with noise and outliers. Compared with D-SVD and ARRBFN, the proposed approach yields much fewer rules and less RMSE values.
基金funded by the National Natural Science Foundation of China(30771518)
文摘In order to isolate genes related to anther development and understand molecular basis of male sterility,cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization（SSH） approach.672 different expressed clones were selected from the fertile disk floret buds.PCR results showed that cDNA inserts were ranged from 100 to 750 bp.303 positive clones screened by dot-blot hybridization were sequenced.273 out of 303 sequenced clones produced readable sequences;these sequences represent 87 non-repetitive sequences.The homology alignment showed that 76 expressed sequence tags（ESTs） had functional annotations in GenBank,the other 11 ESTs without any homology to the known gene.In addition,87 ESTs were divided into 17 groups according to MIPS of Arabidopsis thaliana database.Sequence data from the cDNA library have been deposited with the GenBank under the accession numbers GT067016-GT067085.As an important result in this study,7 genes related to anther development were isolated.Results from semi-quantitative RT-PCR showed 6 genes were expressed only in disk florets of fertile plants compared with that of male sterile plants.These ESTs obtained provide important clues for further isolation and identification of fertility-related genes in Z.elegans.
基金supported by the National Natural Science Foundation of China(30471123,30571206)Natural Science Foundation of Jiangsu Province,China(BK2005421)New Century Excellent Scholar Project of Ministry of Education of China(NCET-07-0042).
文摘Monitoring expression at the transcriptional level is the first essential step for the functional analysis of plant genes. Genes-encoding proteins directly involved in early response to elicitor constitute only a small fraction of all the genes affected by elicitor. Transcriptional responses to various elicitors have been extensively studied in different plants including Nicotiana and Arabidopsis thaliana; however, corresponding data aren＇t available for non-heading Chinese cabbage. To address this problem, we describe a suppression subtractive library-based approach to isolate the plant＇s ESTs up-regulated in the early induction/execution of the HR induced by elicitor PB90 from Phytophthora boehmeriae. According to their putative identification in BLAST searches against the three genome databases, 70 up-regulated genes were classified into 9 parts： some aspect of primary ＇metabolism＇ or ＇energy＇ production; ＇protein synthesis＇ or ＇protein fate＇; cellular communication/signal transduction mechanism; cell fates including Beclin, SPT1, and SPT2; HLA-B and AGO1 which participate in transcription; cellular transport and hypothetical proteins or proteins for which a function has yet to be determined. Seven selected genes such as Beclin, thioredoxin, HLA-B, MAP3K, SPT1, SPT2, and AGO1 were up-regulated induced by PB90, suggesting that the genes may play an important role in PB90-triggered HR.
基金Supported by the National Natural Science Foundation,No. 30371288Beijing Natural Science Foundation,No. 5042024
文摘AIM： To clone and identify human genes transactivated by PSITP5 by constructing a cDNA subtractive library with suppression subtractive hybridization （SSH） technique. METHODS： SSH and bioinformatics techniques were used for screening and cloning of the target genes transactivated by PS1TP5 protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1（-）- myc-his（A）-PS1TP5 and pcDNA3.1（-）-myc-his（A） empty vector, respectively, and SSH technique was employed to analyze the differentially expressed DNA sequence between the two groups. After digestion with restriction enzyme Rsa Ⅰ, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. The tester cDNA was hybridized with driver cDNA twice and subjected to nested PCR for two times, and then subcloned into T/A plasmid vectors to set up the subb-active library. Amplification of the library was carried out with E.. coil strain DH5α. The cDNA was sequenced and analyzed in GenBank with Vector NTI 9.1 and NCBI BLAST software after PCR amplification. RESULTS： The subtractive library of genes transactivated by PS1TP5 was constructed successfully. The amplified library contained 90 positive clones. Colony PCR showed that 70 clones contained 200-1000-bp inserts. Sequence analysis was performed in 30 clones randomly, and the full-length sequences were obtained by bioinformatics technique. Altogether 24 coding sequences were obtained, which consisted of 23 known and 1 unknown.One novel gene with unknown functions was found and named as PSITP5TP1 after being electronically spliced, and deposited in GenBank （accession number： DQ487761）. CONCLUSION： PSITP5 is closely correlated with immunoregulation, carbohydrate metabolism, signal transduction, formation mechanism of hepatic fibrosis, and occurrence and development of tumor. Understanding PSlTP5 transactive proteins may help to bring some new clues for further studying the biological functions of pre-S1 protein.
