近年来,我国的经济高速发展,而人才培养是实现快速发展必不可少的技术手段.因此,高等教育事业也在飞速发展,其中高职护理专业就秉承培养高水平、高素质的护理型专业人才的理念,意在为护理队伍增添新的人才与血液.随着社会不断发展,人们越来越重视培养学生的工匠精神,越来越重视护理专业学生的素质.本文就工匠精神在培养过程中存在的问题,如何培养工匠精神,以及如何将立德树人理念运用在培养工匠精神中等方面进行探析,使其专业知识得以有效发挥,为祖国的护理专业提供一些可用的建议与意见.

案例教学法是新时代下全新的教学方法,与以往的教学方法不同案例教学法是通过典型的案例推动学生在掌握课堂知识点的基础上提升自己解决问题的能力,案例教学法的应用对于提升教师的教学效果是很有帮助的.因此研究案例教学法在教学中的应用非常必要.本文通过分析案例教学法在儿科护理教学中的应用现状,为推动案例教学法在儿科护理教学中的合理应用提出了一些建议.

为了探讨手机APP在高专儿童护理学教学中的应用价值,对我校采取手机APP教学前后的两届学生进行调查和分析,随机在上述两届学生重各抽取了150名学生,将2017届学生(未采用手机APP教学)记为对照组,2018届学生(应用手机APP教学)记为观察组,收集相关数据进行研究和分析,对比开展手机APP辅助教学后学生的学习状况,探讨手机APP在高专儿童护理教学中的应用价值,希望可以找出促进高专学生自主学习,提高高专学校教学质量的有效措施.

SHP2 is encoded by the protein tyrosine phosphatase 11 (Ptpn11) gene. Several gain‐of‐function (GOF) mutations in Ptpn11 have been identified in human hematopoietic malignancies and solid tumors. In addition, the mutation rate for SHP2 is the highest for colorectal cancer (CRC) among solid tumors. The E76K GOF mutation is the most common and active SHP2 mutation; however, the pathogenic effects and function of this mutation in CRC tumor progression have not been well characterized. The Wnt/β‐catenin (CTNNB1) signaling pathway is crucial for CRC, and excessive activation of this pathway has been observed in several tumors. We used Ptpn11E76K conditional knock‐in mice to study this GOF mutation in colitis‐associated CRC (CAC) and used the CRC cell lines HT29 and HCT116 to determine the relationship between SHP2 and Wnt/β‐catenin signaling. Ptpn11E76K conditional knock‐in mice exhibited aggravated inflammation and increased CAC tumorigenesis. In vitro, SHP2E76K and SHP2WT promoted malignant biological behaviors of CRC cells and induced epithelial‐mesenchymal transition (EMT) via the Wnt/β‐catenin signaling pathway. Together, our results showed that SHP2E76K acts as an oncogene that promotes the tumorigenesis and metastasis of CRC.

Objective To construct plasmid for knockout of Kruppel-like factor 4 (KLF4) gene by using CRISPR/ Cas9 gene editing method and examine its impact on biological behavior of human GES-1 cells.Methods Synthesized KLF4 gene targeting sgRNA oligos were inserted into CRISPR/Cas9 plasmid vector of pX459.After transformation,clone isolation,and amplification,DNA sequencing confirmed plasmid DNA,termed pX459-KLF4-sgRNA,was used to transfect human gastric epithelial GES-1 cells in vitro.After transfection,GES-1 cells were first selected in cell-cultured medium containing puromycin,and then the selected GES-1 cells were replated into 100 mm dishes for clonal cell culture.Protein samples extracted from individual cell clones were used for Western blot analysis of KLF4 expression.Biological assays of plate clone formation,MTT cell proliferation,and transwell cell migration were used to compare the cellular behavior changes among GES-1 cells of parental,control clone or clone with KLF4 gene knockdown.Results DNA sequence analysis confirmed that KLF4 targeting sgRNA sequences were successfully constructed into of pX459 vector.Transfection of the resultant vector of pX459-KLF4-sgRNA into GES1 cells led to KLF4 gene knockdown indicated by drastic reduction of KLF4 protein expression,significant biological behavior changes including increased cell proliferation,clone formation,and migration abilities.Conclusion We successfully constructed human KLF4 gene editing plasmid vector based on CRISPR/Cas9 technique,and the biological behavior changes of the established GES-1 cell line with drastic KLF4 knockdown confirmed the tumor suppressive function of KLF4.

Objective To construct a lentivirus vector of microRNA-205(miR-205), and transfect into MB231 cell line, to establish a stable cell line expressing miR-205.Methods Design and synthesis miR-205 primers, the target gene was amplified by PCR, and connect to the lentivirus expression plasmid.Packaging and testing virus drops Lv-miR-205, after the double enzyme digestion of the recombinant plasmid.Infecting MB231 cells with the Lv-miR-205,then detect the expression of miR-205 by Real Time qPCR and access the cell proliferation and migration by MTT assay and wound-healing assay.Results The DNA sequencing prove that the lentivirus vector was constructed successfully.Lv-miR-205 stablized infect MB231 cells, the expression of miR-205 elevated apparently, cell proliferation and migration was inhibited sharply.Conclusion The miR-205 lentiviral expression vector is successfully constructed and the cell line MB231 stably expressing miR-205 is established, which indicate that it could negatively regulate the malignant biological behavior of breast cancer cells, and laid the foundation for the further study of the function and mechanism of miR-205.

Runx2, a member of the Runt domain family, is a well-known master transcription factor for osteoblast differentiation. Runx2 has also been shown to play essential roles during chondrocyte hypertrophy, an important late stage of endochondral ossification linking both bone and cartilage development. To identify the co-factors that may interact with Runx2 together to regulate this critical process, we have performed yeast two-hybrid (Y2H) screening using Runx2 as a bait to screen a cDNA library of hypertrophic chondrocytes. The bait expressing cassette was constructed by fusing Runx2 with the pGBKT7 vector containing the Gal4 DNA binding domain (BD). The Mate & Plate libraries were constructed using pGADT7-Rec and cDNAs derived from hypertrophic chondrocytes enriched limb tissues or hypertrophic MCT cells. After co-transformation of pGBKT7-Runx2 and the cDNA libraries, colonies that grew in nutrition deficient medium were selected and subjected to PCR and sequencing analysis. We successfully identified more than 30 candidate genes, including Lectin-1 (Lgals1), Col1a2, Edf1 and Timp-2. We have performed literature review and bioinformatics analysis of these genes using GenePaint. Most of them show ubiquitous expression with Lgals1 show enhanced expression in hypertrophic chondrocytes. We further performed preliminary expression analysis by quantitative PCR and detected differential expression of these candidate genes in proliferative and hypertrophic MCT cells, with Timp-2 significantly (around 3-fold) and Lgals1 moderately (around 1.5 fold) upregulated in hypertrophic MCT cells. Our results suggest that, candidate gene Timp-2 is very likely to interact with Runx2 and together to play essential function during cartilage development, and possibly its homeostasis.

目的 探讨乳腺癌和癌旁组织中核心结合因子(RunX2)基因的表达差异及其与乳腺癌患者的临床特征和预后的关系.方法 选择75例乳腺癌患者癌组织标本,所有患者术前未经放疗和化疗,采用免疫组化法检测乳腺癌组织和癌旁组织RunX2的表达情况.分析乳腺癌患者RunX2基因的表达与年龄、病理分型、术后分期、淋巴转移、肿瘤大小、雌激素受体(ER)表型、孕激素受体(PR)表型、人类表皮生长因子2(HER2)表型、ki67表达水平以及生存率的关系.结果 75例患者的癌组织中有44例乳腺癌组织高表达RunX2,31例组织低表达RunX2;而癌旁组织仅23例高表达,52例均表达较低.癌组织和癌旁组织中RunX2的表达水平差异有统计学意义(P＜0.05).RunX2基因的表达高低与患者年龄、术后分期、淋巴结转移、病理类型、PR表型、HER2表型无明显相关性,但是与患者的ER表型和ki67表达相关(P＜0.05),并且高表达RunX2患者术后3年的总体生存率较低表达患者差.结论 乳腺癌组织中RunX2基因高表达与患者的ER表型和临床预后呈正相关性,进一步证实了RunX2在乳腺癌发生发展中起到重要作用.

Objective To investigate the effect of the apple peel extract and its active ingredients cyanidin-3-gluco-side(C3G) on malignant breast tumour growth in vivo. Methods C57BL/6 mouse breast cancer model was built to observe the effect of drinking apple peel juice on mouse breast tumour growth. IHC was used to analyze the differ-ential expression of CD31 in the apple peel extract treated group and the control group. HPLC was used to analyze the chemical composition of apple peels extract. The effect of apple peel component C3G on blood vessels formation of mouse breast cancer cell line E0771 was investigated through 3D Matrigel. The effect of C3G on proliferation and migration of SVEC were determined via MTT and Transwell. Results 1% and 2% concentration of apple peel juice inhibited the tumour growth in mouse breast cancer model. The expression of CD31 in apple peel extract trea-ted group was lower than that of control group indicated by IHC staining. C3G was one of the effective components of the apple peel extract through HPLC analysis. C3G inhibited the blood vessels formation of mouse breast cancer cell line E0771 indicated by 3D Matrigel. The migration of SVEC cells was inhibited by C3G. Conclusion The apple peel extract and its effective component C3G inhibit the tumour growth of mouse breast cancer, which is pos-sibly achieved by inhibiting its angiogenesis.

Objective To investigate the biology effect of RunX2 gene after constructing the eukaryotic expression vector of human RunX2 in breast cancer cells.Methods By the recombinant techniques such as PCR amplifica-tion,digestion,ligation,the human RunX2 gene was inserted into the eukaryotic expression vector of pcDNA3.1, and then it was identified by restriction enzyme digestion,RT-PCR,sequencing.Eukaryotic expression vector of human RunX2 gene was transiently transfected into MCF-7 cells.Western blot analysis was applied to detect the expression of RunX2 protein in breast cancer cells MCF-7;MTT assay and wound healing assay were used to detect the cells proliferation and invasion of MCF-7.Results Our result showed that the eukaryotic expression vector of RunX2 was highly expressed RunX2 protein in MCF-7 cells,the cell proliferation and migration abilities were en-hanced in MCF-7 cells with high expression of RunX2 protein.Conclusion RunX2 constructs eukaryotic expres-sion vector and promotes malignant behavior of breast cancer cells.