1,25-Dihydroxyvitamin D3 promotes prostaglandin E1-induced differentiation of HL-60 cells

Human promyelocytic HL-60 cells can be induced by biochemical agents to differentiate in vitro towards divergent types of myelomonocytic cells. It has been reported that prostaglandin E₁ (PGE₁) can induce granulocytic differentiation and that 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) can induce monocytic differentiation. We have now examined the effects of these compounds, both alone and in combination, on HL-60 cell differentiation. PGE₁ (1 g/ml) or 1,25(OH)₂D₃ (10 nM) each inhibited cell proliferation over 48–96 hours of treatment, but combined treatment with both agents was necessary to produce a strong inhibition. The percentage of HL-60 cells that can reduce nitroblue tetrazolium (NBT) (a characteristic index of early monocytic or granulocytic differentiation) increased 13-fold within 72 hours of PGE₁ treatment, and 1,25(OH)₂D₃ produced a fivefold stimulation. However, combined treatment (PGE₁ plus 1,25(OH)₂D₃) produced a dramatic 35-fold increase. HL-60 cells did not produce significant levels of nitric oxide (NO) before 48 hours in culture, and treatment with PGE₁ or 1,25(OH)₂D₃ did not significantly increase cellular NO elaboration over control levels. However, combined treatment produced a striking 12-fold increase over control levels. Similarly, combined treatment was necessary to obtain the maximal time-dependent stimulation of cellular lactate dehydrogenase (LDH) activity (a marker of granulocytic differentiation) as well as acid phosphatase (ACP) activity. During this same period of time, PGE₁, but not 1,25(OH)₂D₃, markedly stimulated cellular claboration of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)-, and 1,25(OH)₂D₃ cotreatment strongly augmented these effects. Thus, combined treatment with 1,25(OH)₂D₃ plus PGE₁ generally augmented the apparent conversion of these cells, producing synergistic (multiplicative) or additive effects. Furthermore, PGE₁ induced within 48 hours the more general phenotypic changes classically associated with the differentiation of these cells: increased expression of chloroacetate esterase (ChAE) (a granulocytic marker), decreases in the nuclear/cytoplasmic ratio (characteristic of development beyond the promyelocyte/myelocyte stage), and major alterations in morphology from floating spherical cells to loosely adherent, elliptical polygons. 1,25(OH)₂D₃ had little effect itself on most of these parameters, but augmented the morphological changes induced by PGE₁ treatment. Within 48 hours, the ability of these cells to reduce the tetrazolium salt WST-1, a general measure of cellular metabolic activity, was increased by PGE₁, but not by 1,25(OH)₂D₃; however, the combination of 1,25(OH)₂D₃ and PGE₁ again produced the strongest stimulation. Similarly, only PGE₁ significantly reduced intracellular ATP levels, but combined treatments produced a more pronounced decrease. In summary, our findings suggest that PGE₁, not 1,25(OH)₂D₃, is sufficient to promote rapid in vitro differentiation of HL-60 cells along the granulocytic pathway; however, the PGE₁-induced conversion of these cells is markedly augmented by cotreatment with 1,25(OH)₂D₃. In addition, these converted HL-60 cells preferentially utilize the glycolytic pathway, rather than the citric acid cycle, for production of ATP, a metabolic characteristic that resembles that described for mature granulocytes.     

胎鼠胃上皮细胞增殖和分化的电镜和免疫组化观察

用电镜和免疫组化的方法观察了15～16天胎龄的胚胎大鼠胃上皮组织在体外培养过程中增殖及分化的情况。结果表明未分化的上皮细胞在培养过程中大量增殖．并分化出粘液细胞、壁细胞等功能细胞。并初步表明氢化可的松可以促进这一分化过程的进行。实验为在体外研究胃粘膜未分化的干细胞的增殖和分化这一重要生理活动建立了方法,所得结果初步提示皮质激素可能会影响胃粘膜干细胞的增殖分化。     

<内经>各学术流派概述

《内经》中存在着多种学术流派,其学派划分方法目前有两种,即以《内经》中的医家名称为纲和以各专题中的不同学说为纲划分。这两种方法均存在不足之处。以阴阳、五行为纲来探讨《内经》中的医学流派应是今后研究的方向之一。     

Chronic treatment with epidermal growth factor stimulates growth of the urinary tract in the rat

Twenty-four male Wistar rats, 8 weeks old, were allocated into three groups and treated with human recombinant epidermal growth factor (EGF) administered subcutancously in doses of 0, 30, and 150 g/kg per day for 4 weeks. Blood sampling was done every 2nd week and urine sampling was done for 2 consecutive days every week. The most striking finding was that the ureters were dose dependently enlarged, due to growth of all layers of the ureteric wall. The urothelium of the bladder showed considerable hyperplasticity with a widening of the basal proliferative compartment and a normal differentiation pattern as observed by the expression of carbohydrate epitopes, characterized with lectinohistochemistry. Blood examination revealed a decrease in blood haemoglobin concentration and a slight increase in serum creatinine concentration in the high-dose group. There were no effects of EGF on the urinary excretion of electrolytes, proteins, and endogenous EGF.     

督脉穴为主治疗帕金森病临床分析

目的探讨针刺配合药物治疗帕金森病的可行性.方法针刺配合口服常规西药剂量的一半与常规西药剂量治疗进行对照研究.结果两组治疗前后差异均有非常显著性意义(P＜0.01),而两组治疗后比较,差异无显著性意义.结论针刺督脉穴为主治疗帕金森病有良好的疗效.     

多种中药成分诱导大鼠骨髓间质干细胞转变为神经元样细胞

目的　体外定向诱导大鼠骨髓间质干细胞 (MSCs)分化为神经元样细胞。方法　通过贴壁法分离大鼠MSCs,体外扩增培养 ,流式细胞仪检测其表面抗原表达 ,中药成分定向诱导MSCs分化为神经元样细胞。光镜下观察细胞形态 ,免疫细胞化学法检测神经细胞特异性抗原标志。结果　大鼠MSCs可通过贴壁法成功分离并可在体外大量扩增。流式细胞仪检测结果显示CD1 4、CD1 1α、CD34、CD38、CD45、CD80、CD86为阴性 ,CD2 9、CD44、CD90、CD1 0 5、CD1 66呈阳性。黄芪、天麻、人参、当归、脑新舒、人参蜂王浆等多种中药诱导 1～ 3h后大部分MSCs转变为神经元样细胞 ,出现胞体和突起 ,免疫细胞化学染色神经元特异性烯醇酶 (NSE)、巢蛋白 (nestin)呈阳性 ,胶质纤维酸性蛋白 (GFAP)阴性。结论　多种传统中药成分及中药制剂体外能诱导大鼠MSCs分化为神经元样细胞     

Down-Regulation of Osteoblastic Cell Differentiation by Epidermal Growth Factor Receptor

The role of epidermal growth factor receptors (EGF-R) in osteogenic cell differentiation was investigated using preosteoblastic MC3T3-E1 (MC3T3) cells and osteoblast-like ROS 17/2.8 (ROS) cells. When cultured in the presence of β-glycerophosphate (GP) and ascorbic acid (AA), MC3T3 cells underwent spontaneous differentiation into osteoblasts which was confirmed as they expressed osteoblast markers such as alkaline phosphatase (ALP), bone sialoprotein (BSP) and osteocalcin (OC). Interestingly, the number of EGF-binding sites decreased during their differentiation into osteoblasts, and the osteogenic protein-1 (OP-1) treatment, which accelerated their differentiation, lowered the number of EGF-binding sites even further. On the other hand, ROS cells with high expression levels of osteoblast markers and no EGF-R, after being transfected with human EGF-R cDNA (EROS cells), expressed numerous EGF-binding sites as well as EGF-R mRNA and protein; in the process, they ceased to express osteoblast markers, indicating their dedifferentiation into osteoprogenitor cells. Both MC3T3 and EROS cells showed increased cell growth in response to EGF, whereas ROS cells did not. These results imply that the EGF/EGF-R system in osteogenic cells has a crucial function in osteoblast phenotype suppression and osteogenic cell proliferation.     

慢性髓性白血病来源的树突状细胞诱导的CTL体外作用研究

 目的　研究慢性髓性白血病来源的树突状细胞的免疫功能。方法　分离慢性期CML患者外周血单个核细胞 (PBMNC) ,用细胞因子GM CSF及IL 4在体外诱导培养DC ,检测细胞表型 ,并观察其诱导的CTL体外抗肿瘤效应。用酶联免疫 (ELISA)方法测定CML DC混合淋巴细胞反应 (MLR)上清液中IL 12及IFN γ的量 ,并与正常DC进行比较。结果　联合GM CSF及IL 4可诱导CML细胞分化为CML DC ,CML DC的CD1a、CD80、CD83表达率均低于正常DC ;CML DC混合淋巴细胞反应上清IL 12及IFN γ含量均低于正常DC ;CML DC能诱导出对自身CML细胞有特异性杀伤作用的CTL。结论　CML DC具有抗原提呈细胞的免疫功能 ,能诱导抗白血病CTL反应。     

金龙胶囊(JLC)对肿瘤细胞诱导分化作用的研究

目的:观察鲜动物药金龙胶囊(JLC)对肿瘤细胞的诱导分化作用.方法:利用体外细胞培养、MTT、NBT染色及吞噬功能测定等方法,观察在不同浓度金龙胶囊作用下人早幼粒白血病细胞系(HL-60)的细胞形态和功能的变化.以维甲酸(RA)作阳性对照.结果:在1×10-6mg/ml浓度以下金龙胶囊在细胞培养中对细胞的存活没有影响.HL60细胞在1×10.和10-9mg/ml作用3或5天后,早幼粒细胞比例减少,中、晚幼粒细胞比例显著增高,与维甲酸的作用相似.金龙胶囊作用后HL-60细胞的NBT还原能力和吞噬功能也有明显升高,与维甲酸作用相似.结论:金龙胶囊可明显促进HL-60细胞形态和功能上分化、成熟,说明金龙胶囊对HL-60细胞具有明显诱导分化作用.     

三氧化二砷诱导人鼻咽低分化鳞癌裸鼠移植瘤分化及相关基因表达的初步研究

 目的　观察三氧化二砷引起鼻咽癌细胞株CSNE 1裸鼠移植瘤分化的形态学特征 ,并初步探讨p5 3,bc1 2和bax在瘤细胞分化和凋亡中所起的作用。 方法　构建移植瘤模型后腹腔注入三氧化二砷(5mg/kg)。光镜及电镜观察移植瘤的形态学变化 ,TUNEL染色计算凋亡率 ,免疫组化法检测 p5 3,bc1 2和bax基因的表达。结果　腹腔注射As2 O3 后 ,裸鼠移植瘤生长抑制 ,抑瘤率为 75 .4 %。瘤组织向成熟分化伴随凋亡细胞增多 ;野生型p5 3及bax高表达 ,bc1 2无变化。结论　As2 O3 能诱导人低分化鼻咽裸鼠移植瘤分化 ,可能与 p5 3及bax高表达相关 ,与bc1 2无关。