FGF4在肺癌细胞体内和体外中表达差异的研究

FGF4已经被证明是癌基因,它涉及肿瘤的生长和转移,为了解FGF4的表达与肿瘤微环境的关系。我们利用FGF4抗体通过免疫组化对一名肺癌患者癌旁,癌组织,癌组织小鼠移植瘤,二次移植瘤以及原代培养的细胞爬片进行FGF4检测,探究其表达差异。通过对比癌组织与对照组（癌旁组织）,FGF4在癌巢中高表达;同样,移植瘤与二次移植瘤的癌巢中与癌旁组织比较,FGF4表达相对较高;但是将癌组织进行原代培养后,免疫组化检测细胞爬片FGF4,发现仅有5%±0.21%的肿瘤细胞表达FGF4,对照蛋白Cytokine作为肿瘤标记物,则在100%的肿瘤细胞中表达。研究提示免疫组化检测到FGF4在体内和体外表达不同,提示肿瘤细胞FGF4的表达与肿瘤微环境调控密切相关,肿瘤微环境对肿瘤细胞的FGF4的调控有着重要作用。     

大白菜游离小孢子培养技术高效体系的研究

以大白菜吉红82、#534DH 群体及#438 群体为试材，研究了利用细胞破碎仪机械提取小孢子与人工挤压提取小孢子对大白菜游离小孢子活力及提取量的影响。结果表明：机械提取替代人工挤压可在短时间内同时提取多份样品，并可快速调整小孢子的培养浓度。收集40 个适期花蕾于5 mL 离心管内，3 000 r·min-1 破碎10 s，将提取的小孢子定容于25 mL NLN-13 培养基，可确保小孢子浓度在适宜培养范围内，并成功诱导胚胎发生。     

Vitamin C enhanced myocardial differentiation of dedifferentiated fat cells

AIM: In order to observe the myocardial differentiation capacity of the dedifferentiated fat (DFAT) cells treated with vitamin C in vitro. METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture. The DFAT cells of passage 3 were used in the study. Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks. The cell morphology was observed under microscope. The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot. RESULTS: Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture. The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence. After treated with neonatal rat heart cell lysate, the DFAT cells became cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure. The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells. The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C. No spontaneous beating cell was observed. CONCLUSION: Vitamin C enhances the differentiation of DFAT cells into cardiomyocyte-like cells.     

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.     

Inlfuence of FSH Treatment on Expression of CDC25A,TSSK3 and P53in Vitro Cultured Sertoli Cells of Calf

CDC25A, TSSK3 and P53 expressionsin vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of...     

生殖细胞早期发生与减数分裂启动调节机制的研究进展

卵巢性生殖疾病发生的主要原因之一是卵子发生过程中减数分裂发生或染色体分离异常。为了更好地进行这些生殖疾病的诊断和治疗,需要充分地了解个体发育的生物学过程中及关键时间点。鉴于此,本文主要阐述了哺乳动物生殖细胞早期发生和减数分裂启动的生理过程,介绍生殖细胞的出现、迁移、性别分化和减数分裂的整个过程。深入了解卵母细胞发生过程将为利用体外发育来源的卵母细胞治疗卵巢性生殖疾病,克服女性不孕、卵巢早衰等重大疾病提供一定的理论支持。     

牛羊胎盘免疫调节因子的提取及对PANC-Ⅰ增殖和迁移影响的比较

胎盘免疫调节因子(placenta immunoregulating factor,PF)是从胎盘提取的小分子多肽,对病毒性疾病、免疫缺陷疾病及恶性肿瘤等具有潜在的临床应用价值.为检测牛(Bos taurus)和绵羊(Ovis aries)胎盘免疫调节因对人(Homo sapiens)胰腺癌细胞(pancreatic cancer cells-Ⅰ,PANC-Ⅰ)增殖与迁移的影响,本研究对从屠宰场获得的牛和绵羊胎盘进行分离,利用双酶水解法制备PF,将质量浓度为100～2 000 ng/mL的牛、羊PF作用于体外培养的PANC-Ⅰ细胞,以3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲酯基)-2-(4-磺苯基)-2H-四唑(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,MTS)法观察细胞增殖抑制率,应用流式细胞技术检测其对PANC-Ⅰ细胞周期的影响.并利用Transwell迁移实验检测添加PF对PANC-Ⅰ细胞迁移能力的影响,同时通过实时荧光定量PCR法检测肿瘤抑制基因(tumor suppressor gene,PS3)和细胞周期依赖性蛋白激酶抑制因子1A基因(cyclin-dependent kinaseinhibitor 1A,P21)的表达情况.结果显示,17.11 g绵羊胎盘制得0.588 mg PF,6.61 g牛胎盘制得0.312 mgPF,胎盘质量与所得PF的质量比约为20 000∶1.在体外培养的PANC-Ⅰ细胞培养液中添加PF,从形态上观察,牛羊PF对PANC-Ⅰ细胞没有明显的影响.绵羊PF处理后PANC-Ⅰ细胞增殖抑制率为9.75％～22.89％,牛PF处理后PANC-Ⅰ细胞增殖抑制率为-3.81％～23.56％,增殖抑制率最显著的绵羊PF浓度为500 ng/mL,牛PF浓度为1 000 ng/mL.绵羊PF在浓度为500 ng/mL时,PANC-Ⅰ细胞的PI和SPF最低,而牛PF各浓度下的胰腺癌细胞增殖指数(proliferation index,PI)和增殖活性(S-phase fraction,SPF)没有明显的变化.同时,牛羊PF对PANC-Ⅰ细胞的迁移能力无显著影响.牛羊PF使PANC-Ⅰ细胞P53表达上调而P21表达下调.上述结果表明,在一定浓度下,PF对体外培养的PANC-Ⅰ细胞增殖有负调控作用,且牛羊胎盘免疫调节因子对胰腺癌细胞的影响不同,可能因为其存在的活性成分有区别.本研究为胰腺癌的治疗提供了新的思路.