Derivation of Putative Porcine Embryonic Germ Cells and Analysis of Their Multi-Lineage Differentiation Potential

Embryonic germ （EG） cells are cultured pluripotent stem cells derived from the primordial germ cells （PGCs） that migrate from the dorsal mesentery of the hindgut to the developing genital ridge. In this study, the morphology of the porcine genital ridge was assessed in embryos harvested on days 22-30 of pregnancy. PGCs from embryos at these stages were cultured to obtain porcine EG cell lines, and EG-like cells were derived from PGCs from embryos harvested on days 24-28 of pregnancy. The EG-like cells expressed Oct4, Sox2, Nanog, SSEA-3, SSEA-4 and alkaline phosphatase （AP）. These cells were able to form embryoid bodies （EBs） in suspension culture and differentiate into cells representative of the three germ layers as verified by a-fetoprotein （AFP）, s-smooth muscle actin （^-SMA）, and Nestin expression. Spontaneous differentiation from the porcine EG-like cells of delayed passage in vitro showed that they could differentiate into epithelial-like cells, mesenchymal-like cells and neuron-like cells. In vitro directed differentiation generated osteocytes, adipocytes and a variety of neural lineage cells, as demonstrated by alizarin red staining, oil red O staining, and immunoftuorescence for neuronal class III [3-tubulin （Tuj 1）, glial fibrillary protein （GFAP） and galactosylceramidase （GALC）, respectively. These results indicate that porcine EG-like cells have the potential for multi-lineage differentiation and are useful for basic porcine stem cell research.

In-Depth Study of Potential-Based Routing and New Exploration of Its Scheduling Integration

Industrial wireless mesh networks(WMNs)have been widely deployed in various industrial sectors,providing services such as manufacturing process monitoring,equipment control,and sensor data collection.A notable characteristic of industrial WMNs is their distinct traffic pattern,where the majority of traffic flows originate from mesh nodes and are directed towards mesh gateways.In this context,this paper adopts and revisits a routing algorithm known as ALFA(autonomous load-balancing field-based anycast routing),tailored specifically for anycast(one-to-one-of-many)networking in WMNs,where traffic flows can be served through any one of multiple gateways.In essence,the scheme is a hybrid-type routing strategy that leverages the advantages of both back-pressure routing and geographic routing.Notably,its novelty lies in being developed by drawing inspiration from another field,specifically from the movement of charges in an electrostatic potential field.Expanding on the previous work,this paper explores further in-depth discussions that were not previously described,including a detailed description of the analogy between an electrostatic system and a WMN system based on precise mapping perspectives derived from intensive analysis,as well as discussions on anycast,numerical methods employed in devising the ALFA scheme,its characteristics,and complexity.It is worth noting that this paper addresses these previously unexplored aspects,representing significant contributions compared to previous works.As a completely new exploration,a new scheduling strategy is proposed that is compatible with the routing approach by utilizing the potential-based metric not only in routing but also in scheduling.This assigns higher medium access priority to links with a larger potential difference.Extensive simulation results demonstrate the superior performance of the proposed potential-based joint routing and scheduling scheme across various aspects within industrial WMN scenarios.

Predicting differentiation potential of human pluripotent stem cells:Possibilities and challenges

The capability of human pluripotent stem cell(hPSC)lines to propagate indefinitely and differentiate into derivatives of three embryonic germ layers makes these cells be powerful tools for basic scientific research and promising agents for translational medicine.However,variations in differentiation tendency and efficiency as well as pluripotency maintenance necessitate the selection of hPSC lines for the intended applications to save time and cost.To screen the qualified cell lines and exclude problematic cell lines,their pluripotency must be confirmed initially by traditional methods such as teratoma formation or by highthroughput gene expression profiling assay.Additionally,their differentiation potential,particularly the lineage-specific differentiation propensities of hPSC lines,should be predicted in an early stage.As a complement to the teratoma assay,RNA sequencing data provide a quantitative estimate of the differentiation ability of hPSCs in vivo.Moreover,multiple scorecards have been developed based on selected gene sets for predicting the differentiation potential into three germ layers or the desired cell type many days before terminal differentiation.For clinical application of hPSCs,the malignant potential of the cells must also be evaluated.A combination of histologic examination of teratoma with quantitation of gene expression data derived from teratoma tissue provides safety-related predictive information by detecting immature teratomas,malignancy marker expression,and other parameters.Although various prediction methods are available,distinct limitations remain such as the discordance of results between different assays and requirement of a long time and high labor and cost,restricting their wide applications in routine studies.Therefore,simpler and more rapid detection assays with high specificity and sensitivity that can be used to monitor the status of hPSCs at any time and fewer targeted markers that are more specific for a given desired cell type are urgently needed.

Basal cell carcinoma stem cells exhibit osteogenic and chondrogenic differentiation potential

Specific cell subpopulations identified as cancer stem cells(CSCs)can be found in basal cell carcinoma(BCC).Generally,CSCs have a marked trans-differentiation potential that could potentially be used in differentiation therapies.However,there are no studies regarding BCC CSCs multipotency.The aim of the study was to analyze the characteristic of CSCs of BCC with emphasis on their differentiation potential upon specific induction.Specific staining and cell morphology were used for differentiation confirmation,along with the expression analysis of osteogenic(ALP,BSP,Runx2,OCN,BMP2),chondrogenic(COL1 and COL2A1),adipogenic(PPAR-γ)and neurogenic(Nestin and MAP2)markers.BCC CSCs differentiated into osteogenic and chondrogenic lineages,as judged by staining and high expression of specific markers(from 2-to 92-fold higher upon induction).Concomitantly with differentiation,the levels of cancer stem cell markers decreased in the cultures.Adipo-differentiation and neuro-differentiation were unsuccessful.In conclusion,BCC CSCs exhibit the capacity to trans-differentiate,a characteristic that may potentially be useful in the development of new strategies for the treatment of aggressive BCCs.

Glypican 4 down-regulation in pluripotent stem cells as a potential strategy to improve differentiation and to impair tumorigenicity of cell transplants

Recent advances in stem cell technologies have opened new avenues for the treatment of a number of diseases still lacking effective therapeutic options.Cell transplantation has emerged as among the most promising clinical intervention for disorders such as injuries,diabetes,liver diseases, neurodegeneration and heart failure （Lee et al., 2013; Forbes and Rosenthal, 2014; Tabar and Studer, 2014）.

Bioinformatics screening of breast cancer-related genes and potential drug research

Objective:To search the the differentially expressed genes between breast cell carcinoma tissues and normal tissues by using bioinformatics technology,and the potential therapeutic drugs for breast cancer were identified,which can provide reference for clinical immune targeted therapy and drug therapy of breast cancer in the future.Methods:"Breast cancer"was searched by using Gene Expression Omnibus(GEO),and GSE79586 chip data was downloaded.The differentially expressed genes in the control group and the breast cancer model group were screened by using bio-communication technology and subjected to GO function analysis,KEGG pathway analysis,differential gene characteristic expression analysis and protein-protein interaction network(PPI)analysis,and the analysis results were further visualized.Prognosis analysis,related function prediction and immune infiltration analysis were performed using the GEPIA,GeneMANIA,and Timer2.0 databases,respectively.Finally,the compounds with potential therapeutic effects on breast cancer are identified through Connectivity Map(CMap).Western blotting and real-time PCR(RT-PCR)were used to verify the core genes and potential therapeutic agents with the highest correlation in vitro.Results:A total of 3916 differentially expressed genes including 1786 up-regulated genes and 2130 down-regulated genes were screened.GO analysis showed that the differential genes were mainly involved in the positive regulation of phosphorylation,secretory vesicles,racemase and epimerase activities.KEGG analysis showed that differential genes were involved in systemic lupus erythematosus,alcoholism,sticky spots,amoebic dysentery Ras signal pathways and other disease pathways.The characteristic expression analysis of differential genes showed that MEK inhibitors,HSP90 inhibitors and signal transduction pathway kinase inhibitors were drugs similar to the differential genes.PPI results showed that H2AFJ,TFF1,GATA3,FOXA1,and CDH1 were core genes related to breast cancer.Two core genes of H2AFJ and TFF1 with the highest correlation were further selected for GEPIA analysis.The results of the analysis showed that the mRNA expression levels of H2AFJ and TFF1 in breast cancer cells were significantly higher than those in normal tissues,and there was a significant correlation with the pathological staging,overall survival rate and disease-free survival rate of breast cancer patients.H2AFJ and TFF1 may be potential prognostic biomarkers for survival of breast cancer patients.The functions of differentially expressed H2AFJ and TFF1 are mainly related to hormone receptor binding,epithelial structure maintenance and epigenetic negative regulation of genes,chromatin tissue involved in negative regulation of transcription,etc.The results of immune infiltration showed that the expressions of H2AFJ and TFF1 had a significant correlation with the infiltration of macrophages,neutrophils,monocytes,CD4+T,CD8+T,B lymphocytes and other immune cells.CMap results showed that compounds such as Gefitinib,Alpelisib,Sorafenib,and Sunitinib had potential therapeutic effects on breast cancer.Western blot and RT-PCR results showed that H2AFJ and TFF1 were significantly overexpressed in breast cancer cells.Gefitinib significantly inhibited the expression of H2AFJ and TFF1 in breast cancer cells(P<0.05,P<0.01).Conclusion:In this study,differentially expressed genes between breast cell carcinoma tissues and normal tissues were screened out by bioinformatics means to further identify key genes and compounds with potential therapeutic effects in the onset process of breast cancer and to further verify the effectiveness of the screened drugs on breast cancer through experiments.It will provide reference for clinical research and development of new drugs against breast cancer in the future in order to develop more effective treatment options.

Differential Proteomics Reveals the Potential Injury Mechanism Induced by Heavy Ion Radiation in Mice Ovaries

In the present study, we used a proteomics approach based on a two-dimensional electrophoresis （2-DE） reference map to investigate protein expression in the ovarian tissues of pubertal Swiss-Webster mice subjected to carbon ion radiation （CIR）. Among the identified proteins, ubiquitin carboxy-terminal hydrolase L1 （UCH-L1） is associated with the cell cycle[1] and that it influences proliferation in ovarian tissues. We analyzed the expression of UCH-L1 and the proliferation marker proliferation cell nuclear antigen （PCNA） following CIR using immunoblotting and immunofluorescence. The proteomics and biochemical results provide insight into the underlying mechanisms of CIR toxicity in ovarian tissues.

A potential integration method for Birkhoffian system

This paper is intended to apply the potential integration method to the differential equations of the Birkhoffian system. The method is that, for a given Birkhoffian system, its differential equations are first rewritten as 2n first-order differential equations. Secondly, the corresponding partial differential equations are obtained by potential integration method and the solution is expressed as a complete integral. Finally, the integral of the system is obtained.

APPROXIMATION OF GENERALIZED BI－AXIALLY SYMMETRIC POTENTIALS WITH FAST RATES OF GROWTH

The paper deals with growth and approximation of solutions (not necessarily entire) of certain elliptic partial differential equations. These solutions are called Generalized Bi-axially Symmetric Potentials (GBSP's). The GBSP's are taken to be regular in a finite hyperball and influence of the growth of their maximum moduli on the rate of decay of their approximation errors in sup norm is studied. The authors obtain the characterizations of the q-type and lower q-type of a GBSP H ∈ HP,0 < R < ∞, in terms of rate of decay of approximation error E.(H,R0), 0 < R0<R <∞.

Fetal cardiac mesenchymal stem cells express embryonal markers and exhibit differentiation into cells of all three germ layers

AIM:To study the expression of embryonal markers by fetal cardiac mesenchymal stem cells(fC-MSC)and their differentiation into cells of all the germ layers. METHODS:Ten independent cultures of rat fCMSC were set up from cells derived from individual or pooled fetal hearts and studies given below were carried out at passages 3,6,15 and 21.The phenotypic markers CD29,CD31,CD34,CD45,CD73,CD90, CD105,CD166 and HLA-DR were analyzed by flow cytometry.The expression of embryonal markers Oct-4, Nanog,Sox-2,SSEA-1,SSEA-3,SSEA-4,TRA-1-60 and TRA 1-81 were studied by immunocytochemistry.The fC-MSC treated with specific induction medium were evaluated for their differentiation into(1)adipocytes and osteocytes(mesodermal cells)by Oil Red O and Alizarin Red staining,respectively,as well as by expression of lipoprotein lipase,PPARγ2 genes in adipocytes and osteopontin and RUNX2 genes in osteocytes by reverse-transcription polymerase chain reaction(RT- PCR);(2)neuronal(ectodermal)cells by expression of neuronal Filament-160 and Glial Fibrillar Acidic Protein by RT-PCR and immunocytochemistry;and(3)hepa- tocytic(endodermal)cells by expression of albumin by RT-PCR and immunocytochemistry,glycogen deposits by Periodic Acid Schiff staining and excretion of urea into the culture supernatant. RESULTS:The fC-MSC expressed CD29,CD73,CD90, CD105,CD166 but lacked expression of CD31,CD34, CD45 and HLA-DR.They expressed embryonal markers,viz.Oct-4,Nanog,Sox-2,SSEA-1,SSEA-3,SSEA-4, TRA-1-81 but not TRA-1-60.On treatment with specific induction media,they differentiated into adipocytes and osteocytes,neuronal cells and hepatocytic cells. CONCLUSION:Our results together suggest that fCMSC are primitive stem cell types with a high degree of plasticity and,in addition to their suitability for cardiovascular regenerative therapy,they may have a wide spectrum of therapeutic applications in regenerative medicine.

Stem cell properties and neural differentiation of sheep amniotic epithelial cells

This study was designed to verify the stem cell properties of sheep amniotic epithelial cells and their capacity for neural differentiation. Immunofluorescence microscopy and reverse transcription-PCR revealed that the sheep amniotic epithelial cells were positive for the embryonic stem cell marker proteins SSEA-1, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81, and the totipotency-associated genes Oct-4, Sox-2 and Rex-1, but negative for Nanog. Amniotic epithelial cells expressed β-Ⅲ-tubulin, glial fibrillary acidic protein, nestin and microtubule-associated protein-2 at 28 days after induction with serum-free neurobasal-A medium containing B-27. Thus, sheep amniotic epithelial cells could differentiate into neurons expressing β-Ⅲ-tubulin and microtubule-associated protein-2, and glial-like cells expressing glial fibrillary acidic protein, under specific conditions.

Effect of Interaction Potential on Laser-assisted e-Ar Scattering

The interaction potentials between electron and atom play an important role in electron- atom scattering. Using three potential models, the absolute differential cross section has been calculated by the second Born approximation theory. Results show that these model potentials are successful in the laser-assisted e-Ar scattering system. The influence of static potential, exchange potential and polarization potential on the absolute differential cross section is also analyzed and discussed.

New models of adipogenic differentiation highlight a cell-autonomous response to temperature

Temperature is a key regulator of brown adipose tissue(BAT)function,acting through central sensory inputs to influence metabolism and energy storage.Although animal models have produced a wealth of information on the pathways,effectors and responses mediating the physiological response of adipose tissue to temperature in vivo,the use of cell culture models now offers evidence of an additional cell-autonomous response to temperature changes,in the absence of neural input.In particular,stem cell models provide new insight into the regulation of adipogenic differentiation and the induction of browning features in vitro.Here the basis for adipogenic responsiveness to low temperature is discussed,together with different human cell models available to outline the benefits of cell-based approaches for future BAT research.

K^- Nucleus Elastic Scattering and Momentum-Dependent Optical Potentials

The K? nucleus differential elastic scattering cross section for 12C and 40Ca at is calculated with three momentum-dependent optical potential models, which are density-dependent, relativistic mean field, and hybrid model, respectively. It is found that the forms of momentum-dependent optical potential models proposed by us are reasonable and gain success in the calculations and the momentum-dependent hybrid model is the best model for the K? nucleus elastic scattering.

花岗伟晶岩分异演化与含矿性评价——从造岩矿物组构视角

高演化稀有金属伟晶岩的成矿过程是复杂和苛刻的,涉及到岩浆-热液过程热历史、岩浆迁移路径与成矿空间、侵位环境的过冷程度以及岩浆组分中稀有金属元素的初始富集等条件。而伟晶岩的分带特征是其形成过程的一种综合体现,不论是伟晶岩区域分带还是伟晶岩内部结构分带,都反映了伟晶岩岩浆结晶分异过程中岩浆的性质与形成环境,而这同时也赋予了造岩矿物在组构上的特殊性,形成遗传代码,对其进行解密,从而能够用于示踪与评估花岗伟晶岩的演化程度与成矿潜力。对于区域伟晶岩群的成矿潜力评估,由于含矿熔体在组分上的跨度较大,随着远离母体花岗岩体距离的增大,伟晶岩群演化程度增加,熔体中铁镁质组分以及Ca、Ba、Sr等碱土金属含量迅速降低,而挥发分、助熔剂组分、碱金属含量在残余熔体中逐渐增加,长石、云母与石英在类别和组构上产生显著变化;其中长石向钾-钠端元演化,且钠长石相对钾长石占据主导地位,云母由黑云母向白云母以及锂白云母转变,石英阴极发光特征与晶体结构及微量元素成分也表现出规律性的变化。受矿物晶格的限制,长石的K/Rb、K/Cs以及云母的K/Rb、K/Cs、Nb/Ta等比值特征能有效区分不同矿化潜力的伟晶岩群。石英的阴极发光特征能够揭示其生长环境与历史,石英的Li、Al、Ti、Ge等微量元素组成可以用来区分不同类型的伟晶岩矿床,并提供关于岩浆演化和成矿过程的重要线索。伟晶岩内部分带的形成主要受成矿熔体规模、过冷度以及成矿空间的封闭性等因素的控制,但在初始熔体成分上可以具有较大的变化。其中强分带型稀有金属伟晶岩记录了完整的岩浆-热液演化过程,不同阶段石英、长石和云母在晶体形态、微观结构以及微量元素组成上都具有显著变化。弱分带型稀有金属伟晶岩(钠长石-锂辉石伟晶岩)的造岩矿物组分在伟晶岩内部变化不大,其受构造控制明显,在空间上与贫矿伟晶岩之间可具有较大的矿物组成差异,表明二者之间存在流动分异过程。而长距离(以更厚的地壳或大型拆离断层为标志)的熔体迁移正是形成超大型伟晶岩稀有金属矿床的有利要素。

沿黄九省区旅游经济发展潜力测度与影响因素

为有效识别沿黄九省区旅游经济发展趋向高质量发展的动力来源和新增长点,运用熵值法、探索性空间数据分析和地理探测器分析法,对沿黄九省区旅游经济发展潜力进行测度,并对其空间分异特征和影响因素进行分析。结果表明:(1)沿黄九省区旅游经济发展综合潜力水平存在显著差距,而相邻省区差距较小,质量效益潜力、动力保障潜力水平除个别省区出现较低和较高值外,其余省区差距较小,各省区经济结构潜力水平均较低且差距较小;(2)沿黄九省区旅游经济发展潜力空间上呈现显著差异和不均衡的分异特征,只有综合潜力和经济结构潜力呈现显著的空间集聚特征,但局部集聚水平较低;(3)上游和中下游发展潜力影响因素差异较大,上游的动力来源主要为质量效益,中下游发展潜力影响因素较全面且只有环境保障因素不显著。该研究可为挖掘沿黄九省区旅游经济发展潜力提供参考。

论海相页岩气富集机理——以四川盆地五峰组—龙马溪组为例

深入探究页岩气富集机理是保障勘探开发高效推进的基础。本研究通过对四川盆地五峰组—龙马溪组页岩气勘探开发实践的系统分析,梳理总结前人研究成果,从生成机理、运移机理、赋存机理和保存机理四个方面对海相页岩气富集机理进行了深入分析,并讨论了深层和常压页岩气的勘探开发潜力。结果表明:在生成机理方面,埋藏史和热演化史控制了页岩生排烃史、生排烃量和现今含气量;页岩气运移机理涉及运移动力、运移相态、运移方式和运移通道四方面内容,页岩气运移主要是烃源岩内的初次运移,同时讨论了初次运移的影响因素;在赋存机理方面,甲烷—页岩间表现出单/多分子层吸附和微孔充填等多种赋存机制,组分润湿性和孔隙有效性是决定甲烷吸附赋存和解吸运移的关键;在保存机理方面,盖层和物性自封闭是主要的保存机理,构造运动引起的裂缝—流体活动是页岩气保存条件遭到破坏的主要原因,流体活动时间和期次研究是页岩气保存条件和含气量定量评价的重要内容。页岩气富集机理的系统分析和创新认识为页岩气勘探开发评价提供了重要依据,建议加强页岩气演化历史全过程的动态评价。结合深层和常压页岩气勘探实践,分析了深层和常压页岩气的成因机制及主要特征,指出了下一步攻关内容及勘探方向。

叠合盆地深层海相油气差异富集历史的动态模拟——以塔河油田下古生界含油气系统为例

以塔里木盆地塔河油田下古生界含油气系统为例,针对叠合盆地深层海相油气复杂的差异富集历史,采用成藏动力学模拟方法进行深层油气的生排烃、运聚和调整改造的历史恢复。研究表明:①塔河油田下寒武统烃源岩的热演化史反映出不同构造带的生排烃过程及其强度具有明显不同,是导致深层油气相态差异的主要原因。②走滑断裂和不整合面等构成的复合输导体系控制深层油气早期运移聚集和后期调整,中寒武统台内膏盐岩阻止深层油气的垂向运聚,致使深层油气呈现明显的“断控”特征,其中北东向走滑断裂带和深层油气运移低势区叠加有利于汇聚成藏,且主要沿北东向走滑断裂带呈串珠状分布。③成藏动态模拟揭示“源-断-缝-膏-保”时空配置控制塔河深层油气的差异富集,奥陶系经历多期充注、垂向运聚、侧向调整改造的成藏历史,深层油气一直处于运移聚集和逸散的动态平衡中。④油气残留量统计显示塔河油田深层奥陶系鹰山组和蓬莱坝组仍具有较好的勘探开发潜力,超深层中上寒武统具有一定的油气资源前景。研究为塔里木盆地深层油气的动态定量评价提供了参考依据,也可以为古老克拉通盆地碳酸盐岩相关油气成藏演化的研究提供借鉴。

电极/碱性聚电解质界面的微分电容曲线和零电荷电位测定

碱性聚合物电解质作为现代碱性氢氧燃料电池的核心组成部分,其单离子导体的特性使得“电极/碱性聚电解质”界面的性质与“电极/溶液”界面有所不同。本文使用微电极,运用循环伏安、电化学交流阻抗以及浸入法等方法,测定了电极/碱性聚电解质界面的微分电容曲线和零电荷电位。该界面的微分电容曲线呈“U”状,且存在局域极小值,该极小值所对应的电位与浸入法测得的零电荷电位数值一致。单离子导体的特性使得“电极/碱性聚电解质”界面在零电荷电位两侧表现出不同的电化学极化行为。

旱柳失水枝条中ABA介导的代谢组响应分析

【目的】ABA是植物干旱胁迫的关键激素,通过代谢组学分析揭示旱柳离体枝条ABA介导的失水响应差异代谢物及其相关通路,为旱柳干旱胁迫响应提供研究基础和理论依据。【方法】以旱柳离体枝条为材料,探究ABA合成抑制剂钨酸钠溶液浸泡后旱柳失水至不同水势枝条木质部代谢组的差异响应。【结果】失水至P50(-1.9 MPa)旱柳枝条代谢组与未失水枝条相比,内源ABA含量显著增加了425.032%,D-果糖等可溶性糖含量与L-谷氨酸等氨基酸含量显著增加。ABA抑制剂处理后内源ABA含量显著降低了79.253%,D-1,5-脱水果糖等可溶性糖含量与苯丙氨酸等氨基酸含量显著降低。植物激素信号转导、类胡萝卜素生物合成和淀粉与蔗糖代谢途径等是旱柳枝条失水响应的关键代谢通路。【结论】旱柳枝条失水后ABA含量显著增加,介导多糖水解过程等降低了细胞渗透势以应对干旱胁迫。