氨基甾体H42649对K562白血病细胞的抑制增殖和诱导分化作用

目的观察氨基甾体H42649对人慢性粒细胞白血病K562细胞系的抑制增殖和诱导分化作用。方法采用液体培养实验,MTT实验,集落培养实验观察10-8～10-4mol/L浓度的H42649对K562细胞增殖能力的影响;采用Wright-Giemsa染色,联苯胺染色和流式细胞术评价10-6mol/L浓度的H42649对K562细胞的诱导分化作用。结果不同浓度的H42649连续作用K562细胞1～5 d后,细胞计数和集落计数明显减少;第5 d处理组的MTT值显著低于对照组,并呈剂量依赖关系;10-6mol/L浓度的H42649对K562细胞作用5d后,联苯胺染色A值升高(P<0.01),形态学观察其趋向成熟分化,流式细胞术检测药物处理4 d后K562细胞膜上CD71表面标记表达阳性率为84%。结论氨基甾体H42649能显著抑制K562细胞的增殖并诱导其向红系分化,提示该药可作为一种新型慢粒白血病细胞的诱导分化剂。     

Skeletal muscle growth defect in human growth hormone transgenic rat is accompanied by phenotypic changes in progenitor cells

Growth hormone (GH) is known to have a pivotal role in the maintenance of skeletal muscle mass. Sarcopenia, the loss of skeletal muscle mass, is a common phenomenon in aging, and it is widely accepted that sarcopenia is largely attributed to age-related decline in GH secretion. In the present study, we tested if human growth hormone transgenic rats (GH-TG rats) whose plasma GH levels are maintained relatively low could be an appropriate model for sarcopenia. Analyses of GH-TG rats revealed that they exhibit skeletal muscle growth defect as well as atrophy of myofibers. The number of myofibers in tibialis anterior muscle was comparable to that of WT rats, while the proportion of type I slow myofibers in tibialis anterior muscle was increased in GH-TG rats after 5 months. Neither increased expression of ubiquitin ligases, MuRF1 and MAFbx, nor indication of apoptotic cell death was observed. Notably, myogenic differentiation potential of skeletal muscle progenitor cells in GH-TG rats was lower than WT rats, and this was accompanied by increased adipogenic potential. These results indicate that GH-TG rats could be a useful model to elucidate the mechanism of sarcopenia induced by reduced GH action and raised the possibility that decreased GH action may cause an alteration of differentiation potential of skeletal muscle progenitor cells.     

类风湿关节炎中白三烯B4间接分化破骨细胞的实验研究

目的探讨在类风湿关节炎（RA）中,白三烯B4（LTB4）能否通过促进核因子Kappa B受体激活剂配体（RANKL）的表达,起到间接分化破骨细胞的作用.方法利用RA滑膜成纤维细胞（RAFLs）和人外周血单核细胞的共培养体系,对照组2.5 ng/ml巨噬细胞集落刺激因子（M-CSF）刺激、实验a组2.5 ng/ml M-CSF＋10-8mol/L LTB4刺激、实验b组2.5 ng/ml M-CSF＋10-8 mol/L LTB4＋100 ng/ml骨保护素（OPG）刺激,培养3周后行抗酒石酸酸性磷酸酶（TRAP）细胞化学染色,通过计数多核性TRAP酶染色阳性的破骨细胞样细胞,比较各组的分化破骨细胞作用.结果对照组几乎没有破骨细胞样细胞,而实验a组则出现较多的破骨细胞样细胞,实验b组则与对照组相似,几乎没有破骨细胞样细胞.结论在RA中,LTB4能够通过促进RAFLs细胞RANKL的表达来间接分化破骨细胞.     

人脐带间充质干细胞定向诱导分化成类神经元的实验研究

目的:采用不同细胞因子定向诱导人脐带间充质干细胞分化成类神经元,为脐带间充质干细胞移植治疗脊髓损伤提供理论依据。方法:采用Ⅱ型胶原酶消化法分离培养人脐带间充质干细胞,采用流式细胞术鉴定细胞。取第5代细胞随机分成A、B、C、D 4组,在A组基础培养基中加入b FGF,B组中加入b FGF和BDNF,C组中加入b FGF、BDNF和BHA诱导培养,D组为对照组,使用含有10%胎牛血清的DMEM-F12培养基培养。诱导2周后采用实时荧光定量PCR检测细胞的nestin、NEFH和GFAP mRNA表达情况,并通过原子力显微镜观察细胞超微结构的的变化。结果:Ⅱ型胶原酶消化法能成功分离人脐带间充质干细胞。通过流式细胞术检测第3代细胞发现,细胞均强表达CD29、CD44和CD105,而CD34、CD45和HLA-DR均未见表达。经定向诱导分化成类神经元后,原子力显微镜观察细胞表面突起增多,实时荧光定量PCR检测显示nestin在A、B、C组均呈阳性表达,NEFH在A、B组呈阳性表达,而GFAP在A、B、C、D 4组均不表达。A、B组n EFH和nestin表达具有显著差异(P﹤0.05)。结论:本实验成功分离培养人脐带间充质干细胞,所培养的细胞扩增迅速,生物学性状稳定;与b FGF单独处理相比,b FGF联合BDNF更能有效诱导人脐带间充质干细胞分化成类神经元。     

大学生自我分化、成人依恋与嫉妒的关系

目的:探讨大学生自我分化、成人依恋与其嫉妒的关系。方法:采用方便取样从湖南某高校选取443名大学生进行大学生自我分化量表、亲密关系体验量表和布伦格尔自我报告嫉妒量表的测试。结果:1男生在情绪反应得分上显著低于女生,而在情感断绝上显著高于女生(t=2.519,-3.493;P0.01)。男生在依恋回避和依恋焦虑得分上均显著高于女生(t=4.269,3.622;P0.01);2自我分化总分与嫉妒呈显著负相关。依恋焦虑与嫉妒呈显著负相关。自我分化总分与依恋回避,依恋焦虑均呈显著负相关(r=-0.285,0.220,-0.346,-0.537;P0.01);3自我分化总分与依恋焦虑均对嫉妒有显著的预测作用(Beta=-0.239,P0.001;Beta=0.113,P0.05);4自我分化总分在依恋焦虑对嫉妒的影响作用中起到完全中介作用,中介效应量为1.199,占总效应的比例为84.49%。结论:提高大学生的自我分化水平,建立安全型依恋以减少嫉妒的消极心理影响,改善大学生人际关系。     

Phenotypic and Functional Analysis of HL-60 Cells Used in Opsonophagocytic-Killing Assay for Streptococcus pneumoniae

Differentiated HL-60 is an effector cell widely used for the opsonophagocytic-killing assay (OPKA) to measure efficacy of pneumococcal vaccines. We investigated the correlation between phenotypic expression of immunoreceptors and phagocytic ability of HL-60 cells differentiated with N,N-dimethylformamide (DMF), all-trans retinoic acid (ATRA), or 1α, 25-dihydroxyvitamin D₃ (VitD₃) for 5 days. Phenotypic change was examined by flow cytometry with specific antibodies to CD11c, CD14, CD18, CD32, and CD64. Apoptosis was determined by flow cytometry using 7-aminoactinomycin D. Function was evaluated by a standard OPKA against serotype 19F and chemiluminescence-based respiratory burst assay. The expression of CD11c and CD14 gradually increased upon exposure to all three agents, while CD14 expression increased abruptly after VitD₃. The expression of CD18, CD32, and CD64 increased during differentiation with all three agents. Apoptosis remained less than 10% until day 3 but increased after differentiation by DMF or ATRA. Differentiation with ATRA or VitD₃ increased the respiratory burst after day 4. DMF differentiation showed a high OPKA titer at day 1 which sustained thereafter while ATRA or VitD₃-differentiated cells gradually increased. Pearson analysis between the phenotypic changes and OPKA titers suggests that CD11c might be a useful differentiation marker for HL-60 cells for use in pneumococcal OPKA.

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甘草抽出物在体外实验中促进精母细胞的分化

目的:明确甘草对精子发生过程中精母细胞分化的影响.方法:采用小鼠睾丸器官培养方式,添加甘草抽出液,进行抗-Rec 8-抗体免疫染色,检测精母细胞的Rec 8的表达,判断其对精母细胞的分化的影响,同时设对照组.结果:发现甘草组和对照组相比有明显的促进精母细胞的分化,且显示浓度依赖性.结论:甘草能促进在小鼠精子发生过程中精母细胞的分化.     

Effect of RAGE and its ligands on CD4+ T cells北大核心CSCD

RAGE (receptor for advanced glycation end products) is a multiligand receptor on the cell surface. Ligand-RAGE interactions activate several signal transduction pathways that propagate cellular oxidative stress and inflammatory response. RAGE expressed on the CD4+ T cells has been identified as a central transduction receptor which affects the activation, proliferation, migration and differentiation of the cells. In addition, blockade of RAGE suppressed the development of multiple immune-related disorders mediated by CD4+ T cells. These studies highlight the importance of RAGE and its ligands for CD4+ T cells. This article briefly reviews the role of RAGE and its ligands on the proliferation, migration and differentiation of CD4+ T cells and summarizes the related research progress.     

巨噬细胞对小鼠诱导多能干细胞向肝祖细胞分化的影响

目的探讨巨噬细胞(Mc)对小鼠诱导多能干细胞(iPSCs)向肝祖细胞(HPCs)分化的影响。方法C57BL/6N小鼠24只,采取腹腔冲洗法获得巨噬细胞,收集上清获得巨噬细胞条件培养基(Mc-CDM)。通过激活素A、骨形态发生蛋白4和成纤维细胞生长因子等诱导小鼠iPSCs向HPCs分化。将HPCs的诱导分为两组,一组使用正常培养基,为对照组(Ctrl组);另一组在诱导D5使用Mc-CDM培养基,为实验组(Mc组)。通过形态学、免疫荧光、Western Blot检测方法,比较正常Ctrl组与Mc组HPCs的形态及相关蛋白表达的差异。计量资料两组间比较采用t检验。结果在体外建立了iPSCs来源的HPCs;HPCs具有向肝细胞分化的潜能。免疫荧光结果显示:与第12天的Ctrl组相比,第12天的Mc组的肝祖细胞特异性蛋白CK19的表达显著增高(0.901±0.072 vs 0.686±0.097,t=-3.093,P<0.05);Western Blot结果显示:与第12天Ctrl组相比,第12天Mc组肝祖细胞相关蛋白CK19的表达显著增高(1.922±0.103 vs 1.448±0.012,t=-7.881,P<0.05);同时,第12天Mc组自噬相关蛋白LC3的表达亦显著增高(1.392±0.042 vs 1.101±0.048,t=-5.978,P<0.05)。结论巨噬细胞可促进小鼠iPSCs向HPCs分化,其机制可能与HPCs细胞自噬水平增加有关。     

Lung growth and maturation in experimental oligohydramnios in the rat

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