Culture and identification of neonatal rat brain-derived neural stem cells

BACKGROUND Timing of passaging,passage number,passaging approaches and methods for cell identification are critical factors influencing the quality of neural stem cells(NSCs)culture.How to effectively culture and identify NSCs is a continuous interest in NSCs study while these factors are comprehensively considered.AIM To establish a simplified and efficient method for culture and identification of neonatal rat brain-derived NSCs.METHODS First,curved tip operating scissors were used to dissect brain tissues from new born rats(2 to 3 d)and the brain tissues were cut into approximately 1 mm^(3)sections.Filter the single cell suspension through a nylon mesh(200-mesh)and culture the sections in suspensions.Passaging was conducted with TrypLTM Express combined with mechanical tapping and pipetting techniques.Second,identify the 5th generation of passaged NSCs as well as the revived NSCs from cryopreservation.BrdU incorporation method was used to detect self-renew and proliferation capabilities of cells.Different NSCs specific antibodies(anti-nestin,NF200,NSE and GFAP antibodies)were used to identify NSCs specific surface markers and muti-differentiation capabilities by immunofluorescence staining.RESULTS Brain derived cells from newborn rats(2 to 3 d)proliferate and aggregate into spherical-shaped clusters with sustained continuous and stable passaging.When BrdU was incorporated into the 5th generation of passaged cells,positive BrdU cells and nestin cells were observed by immunofluorescence staining.After induction of dissociation using 5%fetal bovine serum,positive NF200,NSE and GFAP cells were observed by immunofluorescence staining.CONCLUSION This is a simplified and efficient method for neonatal rat brain-derived neural stem cell culture and identification.

Culture and Identification of Human Amniotic Mesenchymal Stem Cells

Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.

Molecular identification and culture observation on Acrochaete leptochaete (Chaetophoraceae, Chlorophyta) from China

Acrochaete leptochaete （Huber） Nielsen （Chaetophoraceae, Chlorophyta） was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions were used to ascertain the morphological identification of the isolated material. Based on the unialgal culture, asexual reproduction and growth characteristics of A. leptochaete were investigated over wide ranges of temperature and irradiance. Results revealed that asexual reproduction of A. leptochaete could be realized by biflagellate zoospores. The zoospores germinated directly to give self- replicating generations. Zoospore germination was bipolar. A temperature range from 13-21 ℃ and a lower irradiance of 36 μmol/（m2.s） were most favorable for the growth ofA. leptochaete. Thallus organization, an important taxonomic criterion for the genus Acrochaete, was affected markedly by temperature and irradiance. Our results extend the knowledge about the species＇ general biology and its morphological plasticity. For classification and identification of a simple microphytic algae like A. leptochaete, which are traditionally placed in the class Chaetophoraceae, we propose that molecular tools associated with culture observations are applied.

Culture and Identification of Myoblasts Isolated from Duck Embryos

Using embryonic myoblasts to research the formation and de-velopmental mechanisms of skeletal muscle is becoming a research hotspot. This study aimed to establish a method of isolation, culture and identification of my-oblasts in duck embryos. [Method] Pectoral and leg muscle samples were isolated from the embryos of Gaoyou duck at 13 d of hatching, then disassociated with col-lagenase and trypsin and purified via differential adhesion. The isolated cells were cultured in vitro and detected for the expression of Pax7 protein using immunofluo-rescence technique. [Result] Myoblasts were obtained successful y both from pectoral and leg muscles in duck embryos and these cells proliferated strongly and differen-tiated wel . Immunofluorescence staining showed that more than 95% cells could express Pax7 protein. [Conclusion] In summary, we report the successful establish-ment of a complete system for the isolation, purification, identification and culture of myoblasts from duck embryos.

In vitro Culture of Bonamia ostreae and Its Identification

In recent years,bonamiosis has frequently occurred in European areas,which has caused the death of oyster in a wide range and brought enormous economic losses to the breeding industry of oyster. Nowadays,the study on Bonamia sp. is still in the elementary phase. The technology of pathogen’s culture in vitro is the basis for further study on the pathogenesis of Bonamia sp.,its interaction with hosts and the prevention and control of the related disease. In this study,total tissues of oyster identified by PCR were used as culture media to in vitro culture. After one month,they were identified by the method of in situ hybridization. It was found that the results of in situ hybridization were accordant with PCR results. And Bonamia ostreae were detected by in situ hybridization after B. ostreae were cultured for one month. We successfully established a simple and feasible method for in vitro culturing B. ostreae.

Molecular Identification of <i>Campylobacter</i>Species from Positive Cultural Stool Samples of Diarrhoeic Children in Osun State

Death of infants from diarrhoea is a common occurrence in sub-Saharan Africa. This is attributed to unhygienic practices which aid the proliferation of diarrhoea-causing microorganisms. Among these microorganisms, Cam- pylobacter species have been reported as one of the causal agents, Campylobacter spp. are human intestinal pathogens of global importance and their pathogenicity mechanisms are not well understood. This study was designed to investigate the molecular characterisation of Campylobacter gotten from cultural methods in Osun State. Campylobacters isolated were biochemically characterized and biotyped. Confirmation of Campylobacter was done using flaA gene, hippuricase O for Campylobacter jejuni and aspartokinase gene for Campylobacter coli and single locus sequencing glnA gene were performed by PCR. Twenty five samples were amplified by PCR out of 57 Campylobacter strains that were positive for cultural methods from 815 stool samples with diarrhoea and 100 stool samples without diarrhoea. No Campylobacter was isolated from stools of children in the control group. Twenty-five isolates comprising of 18 Campylobater jejuni and 7 C. coli were identified. The nucleotide sequence of the gln A for all the isolated Campylobacter spp. showed 91.0% similarity with the ones in the GenBank. The C. jejuni was classified into biotypes I (44.4%) and II (55.6%) and all C. coli were of biotype I.

Theme Analysis of Everyday Use by Alice Walker——about Afro-Americans' Cultural Identification

By discussing the cultural and historical context of the story and through detailed reviewing of the content,the article tries to deeply analyze the theme of Alice Walker's Everyday Use,and it shows that the writer wants to tell readers two mistakes in Afro-Americans' cultural identification: escaping from the painful history and blind seeking for African culture.

Morphology and Molecular Identification of Dry Fish Fungus Cunninghamella blakesleeana from Small Indigenous Fish “Kachki” Corica soborna (Hamilton 1822) in Bangladesh

The present experiment was conducted to investigate a dry fish fungus, Cunnighamella blakesleeana, which was identified from the infected part of the Corica soborna, locally named as Kachki fish. Mycelium was hyaline, often with granular content, and conidiophores were erected, with verticillate or solitary branches. Zygospores were globose, tuberculate, suspensors equal, smooth, hyaline and heterothallic. Using ITS4 and ITS5 primers, the 740 bp-long ITS region was amplified and sequenced. The ITS region sequences had reciprocal homologies of 98% to 100%. The findings showed that several species of C. blakesleeana fall into the same cluster. It has been determined by molecular data that the fungus we had studied was C. blakesleeana. The maximum mycelial growth (95.33 mm) was observed in the PDA medium, followed by the PSA medium, and the lowest growth (65.50 mm) was measured in the HPA medium in the study of the impact of culture media on the mycelial growth of C. blakesleeana. The influence of temperature on the radial mycelial growth of C. blakesleeana on PDA medium was investigated through five different temperatures. Although pH is a crucial factor in understanding the ecology of spoilage fungus, the highest mycelial growth of C. blakesleeana (88.25 mm) was seen at pH 7, followed by pH 8 and pH 6, while pH 9 was revealed to have the lowest mycelial growth. The outcome suggested that C. blakesleeana thrived in neutral environments.

Cultural Identification in the Context of the Belt and Road Initiative

Culture,as a kind of social and historical phenomenon,is a way of being,and there are complementary characteristics between different cultures.Cultural identification in the context of"the Belt and Road Initiative"(BRI hereafter)manifests itself in the heterogeneous space during the process of constructing cultural subjectivity.Cultural identification,in such a situation,not only embodies the national character of a culture,but also contains the global hybrid.To be specific,cultural identification in cross-cultural transmission,which means overseas acceptance of Chinese culture in this sense,includes self-identity and other-identification.Through cultural dialogue,it finally points to the common shaping of culture beyond boundaries.

Interspecific Hybridization of Brassica campestris × B.oleracea Through Ovary Culture

Using three varieties of Brassica campestris, Hauarad (708), Maoshan-3 (714) and Youbai (715),as the maternal plants and one variety of Brassica oleracea Jingfeng-1 (6012) as paternalplants, crosses were made to produce interspecific hybrids through ovary culture techniques.The ovaries from the cross between B. campestrisB.oleracea (7086012 and 7146012) werecultured and ovary culture was more effective in terms of obtained seeds when ovaries werecultured in vitro at 9 d after pollination (DAP). While for the cross of 7156012, it wasbetter when ovaries in vitro cultured at 12 DAP. Among three cross combinations, the cross of7146012 showed the best response and 43 seeds per ovary were obtained. Among the mediastudied, the ovaries from the cross of 7086012 cultured on MS media supplemented with 3.0 mgL-1 BA0.1 mg L-1 NAA showed better response, and its rate of seeds per ovary reached 44.0%.While the ovaries from the other two crosses (7146012 and 7156012) showed the best responsewhen cultured on B5 media supplemented with 3.0 mg L-1 BA + 0.2 mg L-1 NAA, and the rates of seedsper ovary reached 72.0 and 60.0%, respectively. All seeds obtained from the three crosscombinations were cultured on the MS media supplemented with 1.0 mg L-1 BA + 0.05 mg L-1 NAA,and the seeds from the cross of 7156012 showed the best germination response and thepercentage of germinations reached 66.7%. The regenerated plantlets were obtained from theseseedlings after cultured on the MS media supplemented with 0.05 mg L-1 NAA. Cytological studyshowed that these regenerated plants were all true hybrids of B.campestrisB.oleracea.

Study on the Cross-Cultural Identification of A Community of Shared Future for Mankind

As a kind of“global view”beyond the country and ideology,the concept of a community of shared future for mankind is the core of Xi Jinping’s diplomatic thoughts,and also a Chinese strategy for the future of mankind,which is proposed by China to adapt to the interrelation of human societies with a view of world peace and development.The cross-cultural awareness contained in such concept refers actually to the promotion and abandonment of different cultures in the world.Cross-cultural identification means that the national culture constantly absorbs the positive factors of foreign cultures while discards its negative factors,so as to promote the benign combination of the both and realize the localization of the latter.In the longer term,the most direct mission of the proposal of such concept is to answer the goal of the great rejuvenation of the Chinese nation,and to promote the common revitalization of all countries and resurgence of civilizations.This concept interprets the relationship between China and the world and also integrates the Chinese dream with the world dream.Therefore,the study on cross-cultural identification of all countries should expand international understanding and broad consensus,on the basis of the global perspective of the community of shared future for mankind,and under the principle of“symbiosis,negotiation,collaborative construction,sharing and a win-win situation”.

A novel primary culture method for high-purity satellite glial cells derived from rat dorsal root ganglion

Satellite glial cells surround neurons within dorsal root ganglia. Previous studies have focused on single-cell suspensions of cultured neurons derived from rat dorsal root ganglia. At present, the primary culture method for satellite glial cells derived from rat dorsal root ganglia requires no digestion skill. Hence, the aim of the present study was to establish a novel primary culture method for satellite glial cells derived from dorsal root ganglia. Neonatal rat spine was collected and an incision made to expose the transverse protrusion and remove dorsal root ganglia. Dorsal root ganglia were freed from nerve fibers, connective tissue, and capsule membranes, then rinsed and transferred to 6-well plates, and cultured in a humidified 5% CO_2 incubator at 37°C. After 3 days in culture, some cells had migrated from dorsal root ganglia. After subculture, cells were identified by immunofluorescence labeling for three satellite glial cell-specific markers: glutamine synthetase, glial fibrillary acidic protein, and S100β. Cultured cells expressed glutamine synthetase, glial fibrillary acidic protein, and S100β, suggesting they are satellite glial cells with a purity of > 95%. Thus, we have successfully established a novel primary culture method for obtaining high-purity satellite glial cells from rat dorsal root ganglia without digestion.

The chemical constituents of the tissue culture cells of Daphne giraldii cullus

Three compounds were isolated from the tissue culture cells of Daphne giraldii cullus, their structures were identified as daphneolone （1）, S-（＋）-1-（4-hydroxy-3-methoxyphenyl）-3-hydroxy-5-phenyl-l-pentanone （2）, S-（＋）-1-（4-methoxyphenyl）-3- hydroxy-5-phenyl-l-pentanone （3）, and among them, 2 was a new compound, 3 was a novel natural product.

Advances in In-vitro Culture Technology of Human Fetal Hepatic Stellate Cells

Liver is an important organ for human metabolism and biological conversion. Medical research on hepatic disease clinic, drug metabolism and drug effi- cacy evaluation all needs an in-vitro model of liver as a research platform. Hepatic stellate cells are core cells for occurrence and development of liver fibrosis. Stud- ies at home and abroad deemed that human fetal hepatic stellate cells are an ideal material for the construction of an in-vitro research model for liver fibrosis. With clinical and basic research of liver going deeper, the requirements to quantity and quality of in-vitro models of fetal hepatic stellate cells become higher and higher. The advances in isolation, culture and cryopreservation technique of human fetal hepatic stellate cells were reviewed in this paper.

基于CIS理论的医院文化建设研究

医院文化是在长期的医疗活动中,经过沉淀所形成的一致的价值观念和行为规范,对于全院职工具有导向性、约束性、规范性、凝聚性等作用。而医院文化建设是医院建设发展的中心环节,培养和建设优质的医院文化,有助于医院获得经济和社会效益,更是医院管理者的重要任务。本文分析了企业形象识别系统(Corporate Identity System,CIS)理论体系和文化建设中的精神、制度、行为、物质4个方面的关系,提出了在此理论体系指引下促进医院文化建设的有效策略。

拔尖创新人才培养的理论基础和实践思路

拔尖创新人才的培养是一个现实而又紧迫的话题,但关于什么是“拔尖创新人才”,如何选拔和培养这类人才,尚缺乏成熟的理论准备和实践引导。本文旨在对创新潜能、发展、成就的理论以及对应的识别、培养、文化支持等社会实践做一个系统概要的阐述。首先,在拔尖创新人才的界定上,本文提出了创新的五种形态和创新人才的两种不同类型。第二,在创新人才的识别问题上,本文提出了基于人才动态发展观并结合“总体筛选”和“个体考察”的人才识别模式。第三,在发展和培养问题上,本文提出兼顾成长阶段和领域具体性同时突出“探索未知”和“开放性”的培养模式。第四,在创新成就的文化支持上,本文探讨了如何在创新实践中营造兼容认知多样性和鼓励自由探索的生态环境。第五,本文进一步针对现有教育体制提出了五个实践策略上的着眼点。最后,关于教育实践的政策导向,本文提出了四个基本原则和四种认知误区。

桑树NAC基因家族鉴定与生根粉处理下表达模式分析

【目的】探究桑树NAC基因家族的生物信息学特征,并研究1000 mg·L^(-1)生根粉(ABT)处理对该基因产生的影响,为深入研究桑树NAC转录因子的功能提供依据。【方法】利用生物信息学方法对川桑NAC基因家族进行基因鉴定,并分析其理化性质、保守基序、顺式作用元件、系统进化树、蛋白三级结构及互作关系,并选用‘果桑大十’作为试验材料,经ABT处理后,分析其4个时期(10、20、30、40 d)NAC基因家族的表达模式。【结果】桑树NAC基因家族共有92个成员,分为22个亚家族,亚家族成员广泛分布于细胞核,且具有相似的保守结构。顺式作用元件分析表明,桑树NAC基因家族对激素具有较强的响应能力。蛋白三级结构显示同一亚家族的蛋白空间结构相似,且功能相同,这可能与其进化同源性有关。ABT处理后,NAC基因家族的表达水平整体呈上升趋势,而MnNAC91、MnNAC67、MnNAC28在对照组(CK)中呈现较高的表达水平。【结论】NAC基因家族功能相对稳定,在整个进化过程中无明显变化;ABT可促进NAC基因家族表达,尤其是MnNAC75、MnNAC104、MnNAC7、MnNAC30、MnNAC101、MnNAC83、MnNAC102具有较高的同源性,并与拟南芥的结构功能相似,推测对抗干旱胁迫有一定作用。

黄河流域(山东段)村镇聚落文化景观基因识别指标体系构建

黄河流域(山东段)村镇聚落孕育了丰富独特的文化景观,建立识别指标体系对其文化景观基因进行挖掘整理,对在黄河流域生态保护和高质量发展战略背景下赓续传统文化具有重要意义。结合景观基因理论的研究成果和对国家有关政策法规文件的剖析,在历史景观视角下对黄河流域(山东段)的村镇聚落开展深入的文化景观田野调查、典籍研究等,提出具有针对性的文化景观基因识别指标体系构建逻辑,通过“识别对象—识别因子—识别指标”解析出13项识别因子、40项识别指标,为黄河流域村镇聚落文化景观识别全覆盖提供新的思路与方法。

价值整合、主体认同与制度生成——政府诚信文化建设的再思考

政府诚信是立政之本,现代政府将诚信视为基本的伦理价值追求。作为当前社会诚信规范体系建设的核心环节,政府诚信文化建设一直受到各级政府部门的高度重视,也取得了显著的建设成效。随着信息化时代的来临,政府诚信文化建设面临着更多风险和问题,这对政府诚信文化建设提出了新的要求。为此,应不断整合来源于传统与现实的价值要素,遵从文化主体的理念认同与内化过程,并在融合经验理性与建构理性的生成逻辑基础上创设多方博弈的制度生成情境,以加快促进政府诚信文化建设。

基于改进Yolov5s的增强现实文物识别方法

将增强现实技术应用于数字博物馆的文物展览,有助于拉近参观者与文物的距离,使展览更具趣味性。针对增强现实技术在文物展览应用场景中,增强现实设备采集的文物目标图像背景复杂、文物形状纹理丰富而导致的误检、识别准确率低的问题,提出一种基于改进的Yolov5s的文物识别方法。在Yolov5s网络结构中的骨干网络与颈部网络之间引入了CBAM注意力机制,并在骨干网络中的Bottleneck模块中,使用多头注意力机制替换普通卷积,有效捕获局部信息,降低了无用信息的干扰。为了提高识别网络对于目标文物的边界框定位精度,采用DIoU-NMS方法挑选最优的目标识别框作为最终的预测框。实验结果表明,该方法提高了文物的平均识别精度,比原模型更适用于AR应用文物的目标识别。