Single-cell manipulation by two-dimensional micropatterning

Cells are highly sensitive to their geometrical and mechanical microenvironment that directly regulate cell shape,cytoskeleton and organelle,as well as the nucleus morphology and genetic expression.The emerging two-dimensional micropatterning techniques offer powerful tools to construct controllable and well-organized microenvironment for single-cell level investigations with qualitative analysis,cellular standardization,and in vivo environment mimicking.Here,we provide an overview of the basic principle and characteristics of the two most widely-used micropatterning techniques,including photolithographic micropatterning and soft lithography micropatterning.Moreover,we summarize the application of micropatterning technique in controlling cytoskeleton,cell migration,nucleus and gene expression,as well as intercellular communication.

ANALYTIC EXPRESSION OF MAGNETIC FIELD DISTRIBUTION OF RECTANGULAR PERMANENT MAGNETS

From the molecular current viewpoint,an analytic expression exactly describing magnetic field distribution of rectangular permanent magnets magnetized sufficiently in one direction was derived from the Biot-Savart's law. This expression is useful not only for the case of one rectangular permanent magnet bulk,but also for that of several rectangular permanent magnet bulks. By using this expression,the relations between magnetic field distribution and the size of rectangular permanent magnets as well as the magnitude of magnetic field and the distance from the point in the space to the top (or bottom) surface of rectangular permanent magnets were discussed in detail. All the calculating results are consistent with experimental ones. For transverse magnetic field which is a main magnetic field of rectangular permanent magnets,in order to describe its distribution,two quantities,one is the uniformity in magnitude and the other is the uniformity in distribution of magnetic field,were defined. Furthermore,the relations between them and the geometric size of the magnet as well as the distance from the surface of permanent magnets were investigated by these formulas. The numerical results show that the geometric size and the distance have a visible influence on the uniformity in magnitude and the uniformity in distribution of the magnetic field.

An expert system for predicting shear stress distribution in circular open channels using gene expression programming

The shear stress distribution in circular channels was modeled in this study using gene expression programming(GEP). 173 sets of reliable data were collected under four flow conditions for use in the training and testing stages. The effect of input variables on GEP modeling was studied and 15 different GEP models with individual, binary, ternary, and quaternary input combinations were investigated. The sensitivity analysis results demonstrate that dimensionless parameter y/P, where y is the transverse coordinate, and P is the wetted perimeter, is the most influential parameter with regard to the shear stress distribution in circular channels. GEP model 10, with the parameter y/P and Reynolds number(Re) as inputs, outperformed the other GEP models, with a coefficient of determination of 0.7814 for the testing data set. An equation was derived from the best GEP model and its results were compared with an artificial neural network(ANN) model and an equation based on the Shannon entropy proposed by other researchers. The GEP model, with an average RMSE of 0.0301, exhibits superior performance over the Shannon entropy-based equation, with an average RMSE of 0.1049, and the ANN model, with an average RMSE of 0.2815 for all flow depths.

Two-dimensional Electrophoresis Analysis of Differential Protein Expression in Squamous Carcinoma of the Cervix

Objective： To establish and optimize the two-dimensional gel electrophoresis （2-DE） maps of squamous carcinoma of the cervix and to study the protein difference between squamous carcinoma of the cervix （SCC） and normal cervical tissue. Methods： Using Two-dimensional gel electrophoresis followed by computer-assisted image analysis, the differential proteins between squamous carcinoma of the cervical tissue and normal cervical tissue were compared. Then using matrix-assisted laser desorption/ionization-time of flight mass spectrometry, the differential proteins were identified. Results： The well-resolved and reproducible two-dimensional gel electrophoresis patterns of squamous carcinoma of the cervix tissue and normal cervical tissue were obtained. After silver staining, the average matching ratio of squamous carcinoma of the cervix was 86.1%. There was a good reproducibility of spot position in 2-DE map, with average deviation in IEF direction of 0.95±0.13 mm, while in SDS-PAGE direction it was 1.20±0.18 mm. Ten protein spots were identified by mass spectrometry, some of which were involved in cell proliferation, cell apoptosis, intracellular enzymes, structural proteins, cycle regulation, and tumor occurrence. Conclusion： The differentially expressed proteins provide a fundamental basis for further study of human squamous carcinoma of the cervix and screening of its specific markers.

Spatial distribution patterns of anorectal atresia/stenosis in China:Use of two-dimensional graph-theoretical clustering

AIM:To investigate the spatial distribution patterns of anorectal atresia/stenosis in China.METHODS:Data were collected from the Chinese Birth Defects Monitoring Network(CBDMN),a hospital-based congenital malformations registry system.All fetuses more than 28 wk of gestation and neonates up to 7 d of age in hospitals within the monitoring sites of the CBDMN were monitored from 2001 to 2005.Two-dimensional graph-theoretical clustering was used to divide monitoring sites of the CBDMN into different clusters according to the average incidences of anorectal atresia/stenosis in the different monitoring sites.RESULTS:The overall average incidence of anorectal atresia/stenosis in China was 3.17 per 10000 from 2001 to 2005.The areas with the highest average incidences of anorectal atresia/stenosis were almost always focused in Eastern China.The monitoring sites were grouped into 6 clusters of areas.Cluster 1 comprised the monitoring sites in Heilongjiang Province,Jilin Province,and Liaoning Province;Cluster 2 was composed of those in Fujian Province,Guangdong Province,Hainan Province,Guangxi Zhuang Autonomous Region,south Hunan Province,and south Jiangxi Province;Cluster 3 consisted of those in Beijing Municipal City,Tianjin Municipal City,Hebei Province,Shandong Province,north Jiangsu Province,and north Anhui Province;Cluster 4 was made up of those in Zhejiang Province,Shanghai Municipal City,south Anhui Province,south Jiangsu Province,north Hunan Province,north Jiangxi Province,Hubei Province,Henan Province,Shanxi Province and Inner Mongolia Autonomous Region;Cluster 5 consisted of those in Ningxia Hui Autonomous Region,Gansu Province and Qinghai Province;and Cluster 6 included those in Shaanxi Province,Sichuan Province,Chongqing Municipal City,Yunnan Province,Guizhou Province,Xinjiang Uygur Autonomous Province and Tibet Autonomous Region.CONCLUSION:The fi ndings in this research allow the display of the spatial distribution patterns of anorectal atresia/stenosis in China.These will have important guiding significance for further analysis of relevant environmental factors regarding anorectal atresia/ stenosis and for achieving regional monitoring for anorectal atresia/stenosis.

Protein expression of sensory and motor nerves Two-dimensional gel electrophoresis and mass spectrometry

The present study utilized samples from bilateral motor branches of the femoral nerve, as well as saphenous nerves, ventral roots, and dorsal roots of the spinal cord, to detect differential protein expression using two-dimensional gel electrophoresis and nano ultra-high performance liquid chromatography electrospray ionization mass spectrometry tandem mass spectrometry techniques. A mass spectrum was identified using the Mascot search. Results revealed differential expression of 11 proteins, including transgelin, Ig kappa chain precursor, plasma glutathione peroxidase precursor, an unnamed protein product （gil55628）, gfyceraldehyde-3-phosphate dehydrogenase-like protein, lactoylgfutathione lyase, adenyfate kinase isozyme 1, two unnamed proteins products （gil55628 and gi11334163）, and poly（rC）-binding protein 1 in motor and sensory nerves. Results suggested that these proteins played roles in specific nerve regeneration following peripheral nerve injury and served as specific markers for motor and sensory nerves.

Observation of the Two-Dimensional Distribution of Impurity Line Emissions Using a Space-Resolved EUV Spectrometer in LHD

A space-resolved EUV spectrometer for measuring the one-dimensional distribution of impurity line emissions in Large Helical Device （LHD） has been upgraded to measure two- dimensional distributions of impurity line emissions with an extension of working wavelength range to 30-650% The two-dimensional measurement is performed by scanning the observation chord horizontally. A rectangular plasma region of 520 × 700 mm2 in vertical and horizontal sizes can be observed during a single horizontal scan. The horizontal scan requires a time duration of 5 s at least. The spatial resolution is 10 mm in the vertical direction when a spatial-resolution slit of 0.2 mm in width is adopted. Although a spatial resolution in the toroidal direction is 75 mm, it is a function of CCD exposure time and horizontal scanning speed. Two-dimensional distribution of EUV line emissions from several impurities has been successfully observed for the first time from steady discharges in LHD. In this paper two-dimensional distributions of He II （303.78A）, C V （40.27A）, C VI （33.73A） and Fe XX （132.85A） located at different radial positions are presented with simple analysis on the magnetic field structure of LHD.

Label distribution expression recognition algorithm based on asymptotic truth value

Ambiguous expression is a common phenomenon in facial expression recognition(FER).Because of the existence of ambiguous expression,the effect of FER is severely limited.The reason maybe that the single label of the data cannot effectively describe complex emotional intentions which are vital in FER.Label distribution learning contains more information and is a possible way to solve this problem.To apply label distribution learning on FER,a label distribution expression recognition algorithm based on asymptotic truth value is proposed.Under the premise of not incorporating extraneous quantitative information,the original information of database is fully used to complete the generation and utilization of label distribution.Firstly,in training part,single label learning is used to collect the mean value of the overall distribution of data.Then,the true value of data label is approached gradually on the granularity of data batch.Finally,the whole network model is retrained using the generated label distribution data.Experimental results show that this method can improve the accuracy of the network model obviously,and has certain competitiveness compared with the advanced algorithms.

A Two-Dimensional Brightness Distribution of the Visible Zodiacal Light

By adopting a recently developed semi empirical methodology for removing the atmospheric diffuse light and an improved technique of individual star subtraction, we have newly reduced the distribution of the visible zodiacal light (ZL) brightness from almucantar scan observations of the night sky brightness at Mt. Haleakala, Hawaii. The resulting two dimensional map of the ZL brightness covers the sky over 40°≤λ-λ ⊙≤320° and -40°≤β≤40° and its angular resolution 2°×2°is fine enough to reveal optical counterparts of the structures in the zodiacal emission first found from the IRAS space observations.

Ionic Liquid Assisted Imprint for Efficient and Stable Quasi-2D Perovskite Solar Cells with Controlled Phase Distribution

Although two-dimensional perovskite devices are highly stable,they also lead to a number of challenges.For instance,the introduction of large organic amines makes crystallization process complicated,causing problems such as generally small grain size and blocked charge transfer.In this work,imprint assisted with methylamine acetate were used to improve the morphology of the film,optimize the internal phase distribution,and enhance the charge transfer of the perovskite film.Specifically,imprint promoted the dispersion of spacer cations in the recrystallization process with the assistance of methylamine acetate,thus inhibited the formation of low-n phase induced by the aggregation of spacer cations and facilitated the formation of 3D-like phase.In this case,the corresponding quasi-2D perovskite solar cells delivered improved efficiency and exhibited superior stability.Our work provides an effective strategy to obtain uniform phase distribution for quasi-2D perovskite.

Effect of Polynomial Expressed Distribution Dust Particles for Unmagnetized Two-Ion-Temperature Dusty Plasma

Linear dispersion relation for linear wave and a Kadomtsev-Petviashvili (KP) equation for nonlinearwave are given for the unmagnetized two-ion-temperature cold dusty plasma with many different dust grain species.The numerical results of variations of linear dispersion with respect to the different dust size distribution are given.Moreover,how the amplitude,width,and propagation velocity of solitary wave vary vs different dust size distribution isalso studied numerically in this paper.

TRANSIENT SOLUTION FOR QUEUE-LENGTH DISTRIBUTION OF Geometry/G/1 QUEUEING MODEL

In this paper, the Geometry/G/1 queueing model with inter-arrival times generated by a geometric（parameter p） distribution according to a late arrival system with delayed access and service times independently distributed with distribution {gj }, j≥ 1 is studied. By a simple method （techniques of probability decomposition, renewal process theory） that is different from the techniques used by Hunter（1983）, the transient property of the queue with initial state i（i ≥ 0） is discussed. The recursion expression for u -transform of transient queue-length distribution at any time point n^＋ is obtained, and the recursion expression of the limiting queue length distribution is also obtained.

Two-Dimensional Self-Consistent Kinetic Model for Solenoidal Inductively Coupled Plasma

A two-dimensional self-consistent kinetic model was developed to study the influence of the various factors on the electron energy distribution function. These factors include gas pressure the driving frequency, the radius and length of the inductively coupled plasma equipment, the amplitude of the radio-frequency coil current, and the number of turns of rf coils. The spatial profiles of the rf electric field and power density have also been calculated under the same parameters. Numerical results show that the electron energy distribution functions are significantly modified and the spatial profiles of the rf electric field and rf power density are also demonstrated.

Altered expression of nuclear matrix proteins in etoposide induced apoptosis in HL-60 cells

The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a characteristic morphology change, and the amount of apoptotic cells increased early and reached maximun about 39% after treatment with etoposide for 2 h. Nucleosomal DNA fragmentation was observed after treatment for 4 h. The morphological change of HL-60 cells, thus, occurred earlier than the appearance of DNA ladder. Total nuclear matrix proteins were analyzed by 2-dimensional gel electrophoresis. Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells. Western blotting was then performed on three nuclear matrix acssociated proteins, PML, HSC70 and NuMA. The expression of the suppressor PML protein and heat shock protein HSC70 were significantly upregulated after etoposide treatment, while NuMA, a nuclear mitotic apparatus protein, was down regulated. These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process.

Multicellular tumor spheroids bridge the gap between two-dimensional cancer cells and solid tumors: The role of lipid metabolism and distribution

Previous studies demonstrated that three-dimensional(3D) multicellular tumor spheroids(MCTS) could more closely mimic solid tumors than two-dimensional(2D) cancer cells in terms of the spatial structure, extracellular matrix-cell interaction, and gene expression pattern. However, no study has been reported on the differences in lipid metabolism and distribution among 2D cancer cells, MCTS, and solid tumors. Here, we used Hep G2 liver cancer cell lines to establish these three cancer models. The variations of lipid profiles and spatial distribution among them were explored by using mass spectrometry-based lipidomics and matrix-assisted laser desorption/ionization mass spectrometry imaging(MSI). The results revealed that MCTS, relative to 2D cells, had more shared lipid species with solid tumors. Furthermore,MCTS contained more comparable characteristics than 2D cells to solid tumors with respect to the relative abundance of most lipid classes and mass spectra patterns. MSI data showed that 46 of 71 lipids had similar spatial distribution between solid tumors and MCTS, while lipids in 2D cells had no specific spatial distribution. Interestingly, most of detected lipid species in sphingolipids and glycerolipids preferred locating in the necrotic region to the proliferative region of solid tumors and MCTS. Taken together, our study provides the evidence of lipid metabolism and distribution demonstrating that MCTS are a more suitable in vitro model to mimic solid tumors, which may offer insights into tumor metabolism and microenvironment.

IDENTIFICATION OF DIFFERENTIAL PROTEINS IN UTERINE LEIOMYOMA BY TWO-DIMENSIONAL ELECTROPHORESIS

Objective： To establish and optimize the two-demensional electrophoresis maps of uterine leiomyoma and to study the difference of global protein patterns between uterine leiomyoma and normal myometrium. Methods： Using Two-dimensional electrophoresis followed by computer-assisted image analysis, the differential proteins between uterine leiomyoma and normal myometrium were compared. Results： The well-resolved and reproducible two-dimensional gel electrophoresis patterns of uterine leiomyoma and normal myometrium were established. Totally 1085±108 and 1103±151 protein spots were obtained by using the pH 4-7 IPG strips in uterine leiomyoma and normal myometrium map, respectively, of which 7 spots increased and 15 spots decreased in quantity in uterine leiomyoma compared with normal myometrium. Conclusion： The differentially expressed proteins are useful for studying the mechanism of the cause of uterine leiomyoma.

FEATURE-EXTRACT ANALYSIS OF SERIAL ANALYSIS OF GENE EXPRESSION DATA

Serial Analysis of Gene Expression (SAGE) is a powerful tool to analyze whole-genome expression profiles. SAGE data, characterized by large quantity and high dimensions, need reducing their dimensions and extract feature to improve the accuracy and efficiency when they are used for pattern recognition and clustering analysis. A Poisson Model-based Kernel (PMK) was proposed based on the Poisson distribution of the SAGE data. Kernel Principle Component Analysis (KPCA) with PMK was proposed and used in feature-extract analysis of mouse retinal SAGE data. The computa-tional results show that this algorithm can extract feature effectively and reduce dimensions of SAGE data.

Influence of the channel electric field distribution on the polarization Coulomb field scattering in In_(0.18) Al_(0.82) N/AlN/GaN heterostructure field-effect transistors

By making use of the quasi-two-dimensional （quasi-2D） model, the current-voltage （l-V） characteristics of In0AsA10.82N/A1N/GaN heterostructure field-effect transistors （HFETs） with different gate lengths are simulated based on the measured capacitance-voltage （C-V） characteristics and I-V characteristics. By analyzing the variation of the electron mobility for the two-dimensional electron gas （2DEG） with electric field, it is found that the different polarization charge distributions generated by the different channel electric field distributions can result in different polarization Coulomb field scatterings. The difference between the electron mobilities primarily caused by the polarization Coulomb field scatterings can reach up to 1522.9 cm2/V.s for the prepared In0.38AI0.82N/A1N/GaN HFETs. In addition, when the 2DEG sheet density is modulated by the drain-source bias, the electron mobility presents a peak with the variation of the 2DEG sheet density, the gate length is smaller, and the 2DEG sheet density corresponding to the peak point is higher.

南粳系列超级稻品种灌浆期光合产物的分配特性

【目的】研究南粳系列超级稻灌浆期光合产物转运和分配及相关基因表达量的变化特性及其与对照品种的差异,总结南粳系列超级稻高产的生理优势,为优质高产粳稻的培育提供理论依据。【方法】采用南粳5718、南粳晶谷、南粳3908和南粳5055为研究对象,以淮稻5号为对照,在孕穗期、开花期以及开花后每隔7 d至成熟期测定剑叶光合速率、地上部干物质分配和转运,以及剑叶和籽粒发育不同时期物质转运相关基因的表达量,统计产量差异。【结果】南粳系列超级稻产量和千粒重高于淮稻5号,其剑叶净光合速率在孕穗期和花后28 d显著高于淮稻5号。在物质转运方面,南粳系列超级稻开花后茎叶干重、叶片输出量、输出率和转运率显著高于淮稻5号,其中南粳5718叶片输出量和输出率最高。南粳5718剑叶中与淀粉降解和糖类代谢相关基因(OsSPS1,OsSUT2,OsGWD1)的表达比其他品种启动早,最高表达量也高于其他品种。籽粒中SWEET基因在灌浆早期的蔗糖转运过程中扮演重要角色,中后期OsPK3、OsSUT1、OsSUT2基因在糖类的运输卸载中发挥重要作用,OsAGPL2和OsDPE1基因在中后期淀粉合成中发挥重要作用,南粳5718、南粳晶谷、南粳3908籽粒中与淀粉合成和糖类运输的相关基因在不同时期的表达量显著高于淮稻5号。【结论】南粳系列超级稻较高的产量在物质转运方面主要有以下特征:茎秆和叶片物质积累量多,穗积累量大,叶片和茎秆物质转运率高;叶片中蔗糖代谢和转运相关基因表达量高,有利于蔗糖在源端的合成、装载和转运;籽粒中蔗糖转运和淀粉合成相关基因表达量高,有利于蔗糖在库端的卸载及籽粒中淀粉的合成。

神经介素B及其受体在兔圆小囊中的表达和分布

为检测神经介素B(neuromedin B,NMB)及其受体(neuromedin B receptor,NMBR)基因和蛋白在圆小囊(sacculus rotundus,SR)中的表达模式和分布情况,制备圆小囊组织石蜡切片,通过HE染色,观察圆小囊组织形态;提取圆小囊组织mRNA,采用实时荧光定量PCR(qPCR)检测NMB和NMBR基因在圆小囊中的表达情况;提取圆小囊组织蛋白,采用免疫印迹(Western blot)检测NMB和NMBR蛋白在圆小囊中的表达情况;并采用免疫组织化学法(IHC)检测NMB和NMBR蛋白在圆小囊中的分布情况。结果表明:圆小囊结构与普通动物肠道类似,但具有发达的淋巴组织,且淋巴组织中散在分布着弥散神经内分泌系统细胞;NMB和NMBR mRNA在不同性别兔的圆小囊中均有表达,但NMBR mRNA在公兔圆小囊中的表达显著高于母兔;NMB和NMBR蛋白在不同性别兔的圆小囊中均有表达,但两者在公兔圆小囊中的表达均显著高于母兔;NMB和NMBR蛋白主要分布于圆小囊的淋巴组织及绒毛内,分布情况略有不同。综上,这一研究从形态学和分子生物学方面明确了NMB及其受体在圆小囊中的表达和分布情况,可为后续深入研究NMB及其受体在圆小囊内的功能奠定基础。