Analysis of Amino Acids in a Single Human Red Blood Cell by Capillary Zone Electrophoresis with Intracellular NDA- derivatization and Electrochemical Detection

A novel method for determination of amino acids in individual human red blood cells has been developed. In this method, the derivatization reagents (NDA and CN-) are introduced into living cells by electroporation. After completion of derivatization, the amino acids in a single cell is determined by capillary zone electrophoresis with end-column amperometric detection.

A Novel Real-time Fluorescence Mutant-allele-specific Amplification Method for Rapid Single Nucleotide Polymorphism Analysis

Current methods for single nucleotide polymorphism （SNP） analysis are timeconsuming and complicated. We aimed at development of one-step real-time fluorescence mutant-allele-specific amplification （MASA） method for rapid SNP analysis. The method is a marriage of two technologies： MASA primers for target DNA and a double-stranded DNA-selective fluorescent dye, SYBR Green I. Genotypes are separated according to the different threshold cycles of the wild-type and mutant primers. K-ras oncogene was used as a target to validate the feasibility of the method. The experimental results showed that the different genotypes can be clearly discriminated by the assay. The real-time fluorescence MASA method will have an enormous potential for fast and reliable SNP analysis due to its simplicity and low cost.

An Event Alarm System Based on Single and Group Human Behavior Analysis

Due to the increasing demand for security, the development of intelligent surveillance systems has attracted considerable attention in recent years. This study aims to develop a system that is able to identify whether or not the people need help in a public place. Different from previous work, our work considers not only the behaviors of the target person but also the interaction between him and nearby people. In the paper, we propose an event alarm system which can detect the human behaviors and recognize the happening event through integrating the results generated from the single and group behavior analysis. Several new effective features are proposed in the study. Besides, a mechanism capable of extracting one-to-one and multiple-to-one relations is also developed. Experimental results show that the proposed approach can correctly detect human behaviors and provide the alarm messages when emergency events occur.

Detection and Diagnosis of Gear Fault By the Single Gear Tooth Analysis Technique

This paper presents a procedure of sing le gear tooth analysis for early detection and diagnosis of gear faults. The objec tive of this procedure is to develop a method for more sensitive detection of th e incipient faults and locating the faults in the gear. The main idea of the sin gle gear tooth analysis is that the vibration signals collected with a high samp ling rate are divided into a number of segments with the same time interval. The number of signal segments is equal to that of the gear teeth. The analysis of i ndividual segments reveals more sensitively the changes of the vibration signals in both time and frequency domain caused by gear faults. In addition, the locat ion of a failed tooth can be indicated in terms of the position of the segment t hat deviates from the normal segments. An experimental investigation verified th e advantages of the single gear tooth analysis.

Static and Dynamic Analysis of Lasing Action from Single and Coupled Photonic Crystal Nanocavity Lasers

The single and coupled photonic crystal nanocavity lasers are fabricated in the InGaAsP material system and their static and dynamic features are compared. The coupled-cavity lasers show a larger lasing e^ciency and generate an output power higher than the single-cavity lasers, results that are consistent with the theoretical results obtained by rate equations. In dynamic regime, the single-cavity lasers produce pulses as short as 113 ps, while the coupled-cavity lasers show a significantly longer lasing duration. These results indicate that the photonic crystal laser is a promising candidate for the light source in high-speed photonic integrated circuit.

Main Controlling Factors and Dominant Reservoir Series Analysis of Es_3 in the Qibei Area of Qikou Sag

Based on analysis of geophysical data such as core observation, rock slices identification, physical property, scanning electron microscope, X-ray diffraction, logging data etc., 16 factors of sedimentation, diagenesis, fluid pressure, and their relationships with reservoir physical property were analyzed, and the results indicate sedimentation is the internal factor controlling the reservoir property, diagenesis is the external and final decisive factor and abnormal fluid pressure is an important factor preserving the deep reservoir property. Quantitative characterization of diagenesis indicates that compaction and dissolution are more important than cementation and they respectively cause porosity change of-23.6% and 7.7% and -6.2%. Through optimizing 11 main controlling factors and constructing reservoir evaluation index （REI） according to the hierarchical cluster and principal component analysis, reservoir classification standard was established and reservoirs were divided into four classes. The studies show that Es~ SQ4 consists mainly of class I and II, while Es~ SQ6 is mainly of class III and II; the favorable zone is the north and south slope of Qibei sub-sag and the Liujianfang fault-nose. The successful application of the quantitative and comprehensive evaluation in the Qibei area verifies the advanced, practicable method of less artificial factor is suitable for the low porosity and permeability reservoir.

Path analysis and estimation of additive and epistatic gene effects of barley SSD lines

In the paper presented, 99 single seed descent barley lines as well as their parental forms Roland and Apex were studied. The grain weight components and their interrelations were analyzed using simple coefficients of correlation. The direct and indirect effects of such components on grain weight per plant and 1 000-grain weight were estimated using path analysis. In the 2006 and 2007, the spike length and number of spikelets per spike were the determinants of grain weight. Genetic parameters as additive and epistasis effects were estimated for all studied traits. The results indicate the importance of both additive and epistasis gene effects of number of spikes per plant, grain weight per spike, grain number per plant and grain weight per plant in this study.

Crystal and Molecular Structure Analysis in Knoevenagel Condensation Product of Substituted Napthofuran-2-Carbaldehydes

In the current study, two novel Knoevenagel condensation products of substituted napthofuran-2-carbaldehydes were designed, synthesized and characterized. In order to study the intermolecular interactions of title compounds, single crystals were grown by slow evaporation solution growth technique at room temperature and crystal structure has been determined by single crystal X-ray diffraction technique. Both the molecules crystallize in the monoclinic centrosymmetric space group P21/c with one molecule in the asymmetric unit. In compound [4] the molecules are connected via bifurcated C-H···O=C and C-H···N=C H-bonds and van der Waals interactions forming a layered structure, whereas in compound [5a] the molecular conformation is stabilized via intramolecular C-H···O H-bond and molecule interacts with other molecule generated via 21-screw via bifurcated C-H···O=C along with C-H···N=C H-bonds, which are interacting with nitro- of other molecule generated via same symmetry operation, forming bifurcated C-H···O-N H-bonds, which helps in formation of molecular sheet-like structure.Further, in order to understand the various types and nature of intermolecular interactions in the supramolecular structure Hirshfeld surface analysis and fingerprint plot analysis was carried out.

Source apportionment of PM_(2.5)using PMF combined online bulk and single-particle measurements:Contribution of fireworks and biomass burning

Fireworks(FW)could significantly worsen air quality in short term during celebrations.Due to similar tracers with biomass burning(BB),the fast and precise qualification of FW and BB is still challenging.In this study,online bulk and single-particle measurements were combined to investigate the contributions of FW and BB to the overall mass concentrations of PM_(2.5)and specific chemical species by positive matrix factorization(PMF)during the Chinese New Year in Hong Kong in February 2013.With combined information,fresh/aged FW(abundant ^(140)K_(2)NO_(3)^(+)and ^(213)K_(3)SO_(4)^(+)formed from ^(113)K_(2)Cl^(+)discharged by fresh FW)can be extracted from the fresh/aged BB sources,in addition to the Second Aerosol,Vehicles+Road Dust,and Sea Salt factors.The contributions of FW and BB were investigated during three high particle matter episodes influenced by the pollution transported from the Pearl River Delta region.The fresh BB/FW contributed 39.2%and 19.6%to PM_(2.5)during the Lunar Chinese New Year case.However,the contributions of aged FW/BB enhanced in the last two episodes due to the aging process,evidenced by high contributions from secondary aerosols.Generally,the fresh BB/FW showed more significant contributions to nitrate(35.1%and15.0%,respectively)compared with sulfate(25.1%and 5.9%,respectively)and OC(14.8%and11.1%,respectively)on average.In comparison,the aged FW contributed more to sulfate(13.4%).Overall,combining online bulk and single-particle measurement data can combine both instruments’advantages and provide a new perspective for applying source apportionment of aerosols using PMF.

Nanokit coupled electrospray ionization mass spectrometry for analysis of angiotensin converting enzyme 2 activity in single living cell

Angiotensin-converting enzyme 2(ACE2) is not only an enzyme but also a functional receptor on cell membrane for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2). Here, the activity of ACE2 in single living cell is firstly determined using a nanokit coupled electrospray ionization mass spectrometry(nanokit-ESI-MS). Upon the insertion of a micro-capillary into the living h ACE2-CHO cell and the electrochemical sorting of the cytosol, the target ACE2 enzyme hydrolyses angiotensin II inside the capillary to generate angiotensin 1-7. After the electrospray of the mixture at the tip of the capillary, the product is differentiated from the substrate in molecular weight to achieve the detection of ACE2 activity in single cells. The further measurement illustrates that the inflammatory state of cells does not lead to the significant change of ACE2 catalytic activity, which elucidates the relationship between intracellular ACE2 activity and inflammation at single cell level. The established strategy will provide a specific analytical method for further studying the role of ACE2 in the process of virus infection, and extend the application of nanokit based single cell analysis.

Simultaneous Determination of Five Components in Operculina turpethum

[Objectives]This study was conducted to establish a method for the simultaneous determination of caffeic acid, rutin, ononin, luteolin, and apigenin in Operculina turpethum(L.) S. Manso. [Methods]With ononin from O. turpethum as the internal reference, the five components were separated by HPLC, and the contents of various components were calculated according to the relative correction factors of ononin with caffeic acid, rutin, luteolin, and apigenin. Meanwhile, the calculated results of quantitative analysis of multi-components by single marker(QAMS) were compared with the determined values of the external standard method. [Results] The linear relationship of the five components in their respective ranges was good(r=0.999 9). The average recovery was in the range of 97.48%-101.05%, and the RSD values were in the range of 1.04%-2.71%. The results obtained by QAMS were close to those obtained by the external standard method. [Conclusions] The method is accurate, stable and adaptable, and can be used for the determination of five flavonoids in O. turpethum.

Explore potential biomarkers to provide a theoretical basis for the treatment of liver fibrosis with pirfenidone

Background:Hepatic fibrosis is a common chronic liver disease in clinic,the purpose of our study is to explore potential biomarkers to provide a theoretical basis for the treatment of liver fibrosis with pirfenidone.Methods:We downloaded a gene-sequencing dataset and a single-cell dataset from the GEO database and pirfenidone target genes from three different databases.First,we performed GO,KEGG,and DO analysis on pirfenidone target genes.Then,we grouped the liver tissue sequencing data(GSE162694)in the sequencing data set(N-F0 group and F1-F4 group)and performed gene expression differential analysis on these two groups,weighted gene co-expression network analysis and gene Enrichment analysis.Finally,we intersected the significantly upregulated genes in the F1-F4 group with the pirfenidone target genes and performed PPI network analysis.In order to further explore the expression of both pirfenidone drug target genes and liver fibrosis disease genes(PDLFG)in different immune cells of liver tissue,we used the CD45+cell data in the GSE136103 data set for further analysis.Results:A subnetwork consisting of CDC42,HNF4A,BHLHE40,CCDC71L,NR1H3,TNF,MGLL,GPT,SCD and PLIN1 was screened out,and by analysis,we finally identified the SCD as PDLFG.In single-cell sequencing analysis,we found that SCD was highly expressed in M2-polarized macrophages.Conclusion:SCD may be an important target protein to inhibit the progression of liver fibrosis.

Microfluidic adhesion analysis of single glioma cells for evaluating the effect of drugs

Cancer metastasis is one of the most serious problems for tumor therapy,which is closely related to cell adhesion and deadhesion process.Better comprehension of cell adhesion ability will benefit drug research.Here,a biomimetic microfluidic enzyme digestion method was proposed to gently measure the influence of drugs on cell-matrix adhesion ability at the single cell level.The method can selectively digest the extracellular matrix(ECM)that linked to a single cell,and the trypsin concentration around the cell is relatively uniform and constant,thus the measured cell adhesion strength should be precise.Commercially available anti-cancer agents including 5-fluorouracil(5-FU),actinomycin D(Act D),temozolomide(TMZ)and allicin were evaluated,and the data showed only TMZ and allicin can inhibit cell adhesion significantly under our experiment conditions.The influence of TMZ became more and more obvious as the increase of duration and the effect became prominent only after 6 h adhesion process,which could provide a quick evaluation of whether the drugs are effective to cancer cell(compared with Calcein-AM/PI cell viability test).The adhesion strength of U87 cells decreased when the concentration of TMZ increased,and the effect of TMZ can be effectively inhibited by adding lactic acid to culture medium,which indicated acidic tumor microenvironment could promote drug resistance of tumor cells.Different from conventional evaluation methods which focus on the drugs’influence on cellular viability or metabolism,this work provides a new perspective to study the effect of drugs,which is helpful to enrich the drug evaluation system.

Differential gene network analysis from single cell RNA-seq

Study of gene expression has been arguably the most active research field in functional genomics.Over the last two decades,various high-throughput technologies,from gene expression microarray to RNA-seq,have been widely applied to the wholegenome profiling of gene expression.The commonality of these experiments is that they measure the gene expression levels of"bulk"sample,which pools a large number(often in the scale of millions)of cells,and thus the measurements reflect the average expression

Non-contact mechanical and chemical analysis of single living cells by microspectroscopic techniques

Innovative label-free microspectroscopy,which can simultaneously collect Brillouin and Raman signals,is used to characterize the viscoelastic properties and chemical composition of living cells with sub-micrometric resolution.The unprecedented statistical accuracy of the data combined with the high-frequency resolution and the high contrast of the recently built experimental setup permits the study of single living cells immersed in their buffer solution by contactless measurements.The Brillouin signal is deconvoluted in the buffer and the cell components,thereby revealing the mechanical heterogeneity inside the cell.In particular,a 20%increase is observed in the elastic modulus passing from the plasmatic membrane to the nucleus as distinguished by comparison with the Raman spectroscopic marker.Brillouin line shape analysis is even more relevant for the comparison of cells under physiological and pathological conditions.Following oncogene expression,cells show an overall reduction in the elastic modulus(15%)and apparent viscosity(50%).In a proof-of-principle experiment,the ability of this spectroscopic technique to characterize subcellular compartments and distinguish cell status was successfully tested.The results strongly support the future application of this technique for fundamental issues in the biomedical field.

失能失智老人医疗服务需求满足及其影响因素

目的 调查失能失智老人医疗服务需求满足情况,分析其影响因素,为进一步完善失能失智老人医疗服务提供决策依据。方法 按分层整群抽样方法,在上海市杨浦和奉贤两区抽取60岁及以上失能失智老人402名,采用自制问卷调查。运用单因素分析比较不同状况下失能失智老人医疗服务需求满足情况,运用多因素分析失能失智老人医疗服务需求满足的影响因素。结果 失能失智老人医疗服务需求满足程度偏低,为41.0%。老年人日常生活是否能自理、是否有高血压病、是否需要慢性疾病日常配药和治疗、是否需要老年综合征治疗服务、是否有长期便秘、家人没空送老年人上医院、日常配药便利程度、选择在何处接受药物注射、选择在何处接受食物或尿液插管服务以及社区医生治疗水平不高等是影响失能失智老人医疗服务需求满足的主要因素。结论 失能失智老人医疗服务需求满足度不高,受多因素影响。应进一步加强失能失智老人慢性疾病、老年综合征治疗服务,积极推动多病共治的整合式医疗服务。加强社区和养老机构医疗服务设施建设,方便失能失智老人慢性疾病日常管理和医疗护理服务,不断提高失能失智老人医疗服务需求满足程度。

Sequence Information on Simple Sequence Repeats and Single Nucleotide Polymorphisms through Transcriptome Analysis of Mungbean

Mungbean （Vigna radiata （L.） Wilczek） is a unique species in its ability to fix atmospheric nitrogen, with early maturity, and relatively good drought resistance. We used 454 sequencing technology for transcriptome sequencing. A total of 150 159 and 142 993 reads produced 5 254 and 6 374 large contigs （〉_500 bp） with an average length of 833 and 853 for Sunhwa and Jangan, respectively. Functional annotation to known sequences yielded 41.34% and 41.74% unigenes for Jangan and Sunhwa. A higher number of simple sequence repeat （SSR） motifs was identified in Jangan （1 630） compared with that of Sunhwa （1 334）. A similar SSR distribution pattern was observed in both varieties. A total of 8 249 single nucleotide polymorphisms （SNPs） and indels with 2 098 high-confidence candidates were identified in the two mungbean varieties. The average distance between individual SNPs was -860 bp. Our report demonstrates the utility of transcriptomic data for implementing a functional annotation and development of genetic markers. We also provide large resource sequence data for mungbean improvement programs.

A dCAPS marker developed from a stress associated protein gene TaSAP7-B governing grain size and plant height in wheat

Stress associated proteins（SAPs） are the A20/AN1 zinc-finger proteins which confer to abiotic stresses in plants. In this study, TaSAP7-B, including two AN1 domains, was isolated from B genome of wheat（Triticum aestivum L.）. Sequencing analysis on TaSAP7-B illustrated one In Del（insertion-deletion） and one SNP（single nucleotide polymorphism） in the promoter region while no diversity was observed in the coding region. On the basis of SNP in the promoter region（–260 bp）, a dCAPS（derived cleaved amplified polymorphic sequences） marker SNP-260 was developed for TaSAP7-B. Using a natural population consisting of 262 wheat accessions, significant associations were detected between the marker SNP-260 and agronomic traits, such as plant height（PH）, peduncle length（PL）, length of penultimate internode（LPI）, number of spike per plant（NSP）, and 1 000-grain weight（TGW）. Two genotypes were identified using marker SNP-260 in the natural population. Among them, the genotypes possessing C allele exhibited a higher TGW and shorter PH than the T genotypes. Hence, base C was considered as the superior allele. The dCAPS marker of TaSAP7-B can be instrumental for marker-assisted selection for high grain size and short plant height.

Multiscale analysis of single- and multiple-pulse laser-induced damages in HfO_2/SiO_2 multilayer dielectric films at 532 nm

Nanosecond single- and multiple-pulse laser damage studies on HfOffSiO2 high-reflection （HR） coatings are performed at 532 nm. For single-pulse irradiation, the damage is attributed to the defects and the electric intensity distribution in the multilayer thin films. When the defect density in the irradiated area is high, delami- nation is observed. Other than the 1064 nm laser damage, the plasma scalding of the 532 nm laser damage is not pits-centered for normal incidence, and the size of the plasma scalding has no relation to the defect density and position, but increases with the laser fluence. For multiple-pulse irradiations, some damage sites show deeper precursors than those from the single-shot irradiation due to the accumulation effects. The cumulative laser- induced damages behave as pits without the presence of plasma scalding, which is unaffected by the laser fluence and shot numbers. The damage morphologies and depth information both confirm the fatigue effect of a HfO2/SiO2 HR coating under 532 nm laser irradiation.

Mass spectrometry profiling of single bacterial cells reveals metabolic regulation during antibiotics induced bacterial filamentation

Bacterial antimicrobial resistance(AMR)is a severe threat to global health and development.Under the stimulation of antibiotics,bacterial cells can undergo filamentation and generate daughter cells with stronger AMR.The current research on bacterial AMR mechanism is mainly conducted with a population of cells.However,bacterial cells exhibit heteroresistance,making the study at population level not reliable.Herein,we developed single bacterial cell metabolic profiling by mass spectrometry(MS)to study bacterial AMR at single-cell level.By utilizing a microprobe controlled by a microoperation platform,single filamentous extended spectrum beta-lactamase(ESBL)producing Escherichia coli(ESBL-E.coli)cells generated by ceftriaxone sodium stimulation can be extracted and spray-ionized for MS analysis.Heterogeneous among ESBL-E.coli cells under the same antibiotic stimulus condition was observed from mass spectra as well as cell morphology.The metabolic profiles by MS of different individual cells can be clustered into subgroups well in accordance with bacterial cell length.Metabolic pathways including arginine and proline metabolism,as well as cysteine and methionine metabolism were disclosed to play an important role in the bacterial SOS-associated filamentation against antibiotics.The microprobe electrospray ionization-MS-based single bacterial cell analysis method is promising in the study of various bacterial AMR mechanism and can reveal the heterogeneity of bacterial AMR from-cell-to-cell.