Long-term culture of murine erythropoietic progenitor cells in the absence of an adherent layer

Replication of multipotential stem cells in long-term murine bone marrow cell culture is known to depend on the development of an adherent stromal cell layer. In these conditions, restricted haematopoietic progenitor cells have also been generated for up to several months1-3. However, maturation is observed only in the granulocyte/macrophage and megakaryocyte lineages; erythropoiesis appears to be blocked at the earliest burst-forming unit (BFU-E) stage. Addition of exogenous erythropoietin (Epo) or anaemic mouse serum results in full erythropoietic maturation, but it is transient. We describe here a culture system in which production of erythropoietic progenitor cells can be maintained for over 6 months in the absence of an adherent stromal layer and in the absence of added Epo, but in the presence of pokeweed mitogen-stimulated spleen cell conditioned medium (PWSCM). The data indicate that restricted erythroid progenitor cells exist which are capable of extensive self-renewal.     

Long-term proliferation of mouse primordial germ cells in culture.

Primordial germ cells (PGCs) are first identifiable as a population of about eight alkaline phosphatase-positive cells in the 7.0 days postcoitum mouse embryo. During the next 6 days of development they proliferate to give rise to the 25,000 cells that will establish the meiotic population. Steel factor is required for PGC survival both in vivo and in vitro and together with leukaemia inhibitory factor stimulates PGC proliferation in vitro. In feeder-dependent culture, PGCs will proliferate for up to 7 days, but their numbers eventually decline and their proliferative capacity is only a fraction of that seen in vivo. Here we report a further factor that stimulates PGC proliferation in vitro, basic fibroblast growth factor (bFGF). Furthermore, bFGF, in the presence of steel factor and leukaemia inhibitory factor, stimulates long-term proliferation of PGCs, leading to the derivation of large colonies of cells. These embryonic germ cells resemble embryonic stem cells, pluripotent cells derived from preimplantation embryos, or feeder-dependent embryonal carcinoma cells, pluripotent stem cells of PGC-derived tumours (teratomas and teratocarcinomas). To our knowledge, these results provide the first system for long-term culture of PGCs.     

双氯芬酸对Jurkat细胞的抑制作用

目的:探讨双氯芬酸体外对淋巴瘤细胞的抑制作用及机制。方法:分别以双氯芬酸10、30、100μg·mL-1体外处理T细胞淋巴瘤细胞系(Jurkat细胞),采用流式细胞仪和细胞生长曲线观察双氯芬酸对Jurkat细胞的影响。结果:双氯芬酸在体外显著抑制Jurkat细胞的增殖,其抑制作用随着剂量的加大而增强,和空白对照组比较有显著性差异。经流式细胞仪分析显示,双氯芬酸与Ju rkat细胞共同培养,可使细胞凋亡率显著升高。结论:双氯芬酸体外能显著抑制Jurkat细胞的增殖,此作用可能与诱导淋巴瘤细胞发生凋亡有密切关系。     

Extensive proliferation of mature connective-tissue type mast cells in vitro

There are two phenotypically distinct subpopulations of mast cells in rodents: connective tissue-type mast cells (CTMC) and mucosal mast cells (MMC). These populations differ in their location, cell size, staining characteristics, ultrastructure, mediator content and T-cell dependency. Several investigators recently reported a further subclass of mast cells which arise when normal mouse haematopoietic cells are cultured with interleukin-3 (IL-3); IL-3 is an activity similar or identical to mast-cell growth factor, histamine-producing factor, or P-cell stimulating factor. These cultured mast cells are in many ways similar to MMC; they stain with Alcian blue but not safranin, contain chondroitin sulphate E proteoglycan rather than heparin proteoglycan and have relatively low histamine content, as do MMC. Although proliferation of MMC is known to be T-cell dependent in vivo and thought to be IL-3-dependent in vitro, the factors on which CTMC proliferation depends remain elusive. Here we show that mature CTMC purified from mouse peritoneal cells can proliferate in vitro in methylcellulose culture and maintain the appearance and function of CTMC. We also present evidence that mature CTMC cannot proliferate in the presence of pure IL-3 alone.     

选择性COX-2抑制剂尼美舒利对白血病K562细胞增殖的影响

目的:观察选择性COX-2抑制剂尼美舒利对人类骨髓白血病K562细胞增殖和凋亡的影响。方法:分别以尼美舒利25、50、100μg.mL-1体外处理骨髓白血病K562细胞,采用MTT比色法、流式细胞仪分析技术及DNA片段凝胶电泳等方法,检测尼美舒利对K562细胞增殖和凋亡的影响。结果:尼美舒利剂量依赖性地抑制K562细胞增殖,其中等剂量组(50μg.mL-1)的抑制作用强于阿霉素组(25μg.mL-1);尼美舒利可诱导细胞凋亡,使细胞阻滞于G0/G1期,S期细胞明显减少,其诱导细胞凋亡的作用也呈剂量依赖性(剂量由低至高,凋亡率分别为(3.4±2.8)%,(16.8±1.8)%和(42.7±2.5)%;DNA条带分析也进一步证实了尼美舒利的促凋亡作用。结论:尼美舒利在体外可显著抑制K562细胞增殖,该作用可能与其干扰细胞周期、诱导细胞凋亡有关。     

阿维A对HaCaT细胞增殖的影响

目的:探讨阿维A对角质形成细胞(HaCaT)增殖的影响及其发挥作用的适宜浓度.方法:分别以浓度为1×10-7mol/L、1×10-6mol/L、5×10-6mol/L阿维A作用于HaCaT细胞,用四氮唑盐类化合物(MTS)比色法检测每组12、24、48和72 h HaCaT细胞的增殖情况.结果:与空白对照组相比,大于或等于1×10-6mol/L阿维A可以抑制HaCaT细胞增殖,并且在作用24 h后随着阿维A浓度的增加,其对HaCaT细胞增殖的抑制作用增强.结论:阿维A抑制HaCaT细胞增殖适宜浓度为5×10-6mol/L.     

扰动酉系综的1-级相关函数

在随机矩阵理论中,1-级相关函数 R¹_nβ(x)（β一般称为Dyson指标）可以用来刻画在 x 附近能够发现能级的分布密度。直到现在,它的极限行为仍然受到许多数学家和物理学家的关注。在酉系综情形下（即 β=2）,1-级相关函数 R¹_n2(x) 与经典多项式理论中的权函数 μ(x) 密切相关。文中,为叙述上简单起见,只考虑Gauss酉系综的情形。可以发现,在弱收敛意义下,给权函数 μ(x) 以“好的”而非平凡的乘积因子,1-级相关函数 R¹_n2(x) 的极限行为不受干扰。     

基于小波的扰动流场下细胞形态图像

研制带后向台阶的流动腔实验装置,将内皮细胞成功地种植到流动腔底部,并在扰动流场作用下加以培养;应用基于多分辨率分析的二维小波图像压缩以及图像降噪与压缩方法,对图像进行了有效的降噪与压缩.流动腔内位于第1、2和3位置的细胞在扰动流场剪切应力作用12 h后,摄取它们的图像并进行了灰度直方图分析.分析表明,细胞形态与其所受的剪切应力有密切的关系,即细胞沿剪应力的方向发生拉伸现象,剪应力的幅值越大,拉伸越显著.反映在直方图上,细胞承受剪应力的增加导致其图像的直方图中心向灰度值低端移动.研究结果对流动腔实验装置的集成化、微型化和图像的实时处理与显示具有一定的价值.     

凶险性前置胎盘127例临床分析

目的:探讨凶险性前置胎盘的围手术期的高危因素及应对策略.方法:对127例凶险性前置胎盘的病例产前出血、术中出血、围手术期出血、子宫切除率、及其他母儿严重并发症进行分析.根据前置胎盘类型分为:中央性前置胎盘组与部分行前置胎盘组.根据术中胎盘剥离情况分为:胎盘植入组、胎盘粘连组及正常剥离组.结果:(1)胎盘植入率:中央组为49.5%,明显高于部分组胎盘植入率23.1%(P0.05).(2)围手术期出血:中央组平均为1 800 m L,高于部分组平均1 128 m L(P0.01).胎盘植入组平均2 636 m L,胎盘粘连组平均1 342 m L,胎盘正常剥离组平均822 m L,各组间均有统计学差异(P0.01).(3)输血情况:胎盘植入组有98.2%患者术后接受输血治疗,平均输血量2 255 m L,胎盘粘连组有88.2%患者接受输血,平均1 267 m L,正常剥离组有53.7%患者接受输血,平均1 165 m L,胎盘植入组明显高于粘连组(P0.05)及正常剥离组(P0.01),而粘连组与正常剥离组比较无统计学差异(P0.05).(4)子宫切除情况:中央组为34.7%,高于部分组15.4%(P0.05).胎盘植入组为62.5%,明显高于胎盘粘连与正常联合组5.6%(P0.05).(5)预后:产妇有1例术中出现心脏骤停经抢救预后良好,1例新生儿畸形死亡,其余母儿预后均良好.结论:凶险性前置胎盘胎盘植入率高,合并中央性前置胎盘者,出血是最主要的风险,大量输血准备及果断行子宫切除术是手术成功的关键.     

扰动近地层湍流互谱特征研究

运用国际能量平衡实验(EBEX-2000)的湍流、净辐射和温度梯度资料, 对近地层8.7 m和2.7 m两个高度的湍流互谱结构、温度廓线和湍流通量的特征进行了分析, 重点研究了逐块灌溉所致的扰动近地层内大涡与局地湍流的相互作用以及大涡对地表通量的影响。研究结果显示, 低频湍流互谱的峰值频率在两个高度趋于一致, 基本遵循外层标度律(OLS)模型; 高频湍流符合局地各向同性湍流理论。由于大涡的影响, 湍流互谱低频能量显著增强, 且该增强作用在8.7 m高度比2.7 m高度更为突出。湍流谱的低频部分出现多峰值, 峰值频率对应的涡旋尺度与热力非均匀性的尺度有较好的对应关系, 主要的3个涡旋尺度分别为800, 400和200 m。大尺度涡旋扰动对局地湍流的影响: 上层大于下层, 不稳定时大于稳定时。     

扰动压缩条件的点列收敛性问题

本文研究完备度量空间X中满足ρ（Xnrxw＋1）≤LP（xn＋1,Xn）＋sn的点列{xn}收敛性问题,其中L∈（0,1）为常数,εn非负是无穷小量称为扰动,文中的主要结论是：点列{Xn}的收敛性由扰动εn决定,即当幂级数岛∑n=1^∞ εnxn的收敛半径R〉I/L时,点列{xn}收敛,特别地,当R〉1时,点列收敛;而时,{xn}敛散性不能确定。     

Expression of human proteoglycan in Chinese hamster ovary cells inhibits cell proliferation

In studying the functional role of an extracellular matrix proteoglycan, decorin, we have made observations that suggest a role for this proteoglycan in the control of cell proliferation. Extracellular matrices are made up of different combinations of collagens, elastin, hyaluronic acid, proteoglycans and various glycoproteins such as fibronectin. Most of these components can interact with cells, and much of the control of cell adhesion, migration and differentiation appears to be mediated by these interactions. Earlier studies have also attributed growth-regulatory activities to intact extracellular matrices, but the individual molecules responsible for these effects have not been characterized. We report here that Chinese hamster ovary (CHO) cell lines expressing human decorin from a stably transfected complementary DNA construct form a more orderly monolayer and grow to a lower saturation density than control cells lacking decorin. The extent of the morphological changes correlates with the level of decorin expression, and the saturation density is inversely proportional to it. The reduction in the saturation densities of the cell lines with the highest expression of decorin is more than 50%. These results reveal a novel growth inhibitory mechanism which may be related to contact inhibition of cell proliferation.     

2-ME抑制人子宫内膜癌细胞株增殖的研究

目的　探讨2-甲氧雌二醇(2-ME)对人子宫内膜癌细胞株KLE细胞体外增殖和凋亡的抑制作用.方法选用人子宫内膜癌细胞株KLE进行体外培养,实验组加入不同浓度2-ME的培养液,对照组不含2-ME.用四甲基偶氮唑蓝(MTT)比色法观察2-ME对人子宫内膜癌细胞株KLE增殖的抑制作用;药物作用后的克隆形成实验;电子显微镜(电镜)观察细胞形态变化;流式细胞仪(FCM)观察细胞的凋亡率及细胞周期的变化.结果2-ME浓度为10.0～50.0μM时,明显抑制KLE细胞的增殖(P<0.01),并具有时间依赖性和剂量依赖性.2-ME作用后G0/G1期细胞增加,并伴随G0/G1期细胞的增加,出现细胞凋亡峰和凋亡率的升高(P<0.05).电镜下观察到KLE细胞染色体边集、核固缩、凋亡小体.结论2-ME对人子宫内膜癌KLE细胞株增殖有抑制作用,并能促进其凋亡.     

Stability of Second Order Differential Equation Disturbed with Delays

IntroductionThe stability of the zero solution for the second ordernonlinear differential equations disturbed with delays·x·(t) +p(t) x·(t) +q(t)x(t) +f(t , xt) =0,t≥τ(1)was considered in the paper , wherep( t)andq( t)arecontinuous on[τ,+∞) ,f ( t ,)is continuous on[τ,+∞)×C, C≡C([-r ,0] , R),for||≡sup|(θ)|,∈C, xt∈Cis defined byxt(θ)=x(t+θ) ,θ∈[-r ,0] .Iff≡0,the equation (1) becomes the ordinary differentialequation·x·(t) +p(t) x·(t) +q(t)x(t) =0. (2)The zero so…     

一类受扰时滞系统的滑模观测器设计

针对一类时滞系统,首先考虑时滞系统不包含外部干扰时观测器的设计问题,其次考虑了包含外部干扰的时滞系统的观测器设计问题·所研究的时滞系统中包含的干扰项由两部分组成,一部分是满足全局李普希兹条件的非线性项,另一部分为有界不确定项,并且系统时滞是可变的·基于李亚普诺夫稳定性理论利用滑模控制使得误差系统镇定,并将该稳定性问题转化为一类线性矩阵不等式的求解问题,使得构造的滑模观测器能够观测原系统·最后给出仿真实例,说明所设计的观测器是有效可行的·     

佛手水煎剂对RAW264.7癌细胞增殖的影响

为了观察佛手水煎剂对RAW 264.7癌细胞增殖的影响,用生药量0.625,1.250,2.500和5.000 mg/mL的佛手水煎剂处理RAW 264.7癌细胞,采用四甲基偶氮唑蓝(MTT)法测定细胞活力、台盼蓝排斥法测定细胞存活率、吖啶橙/溴化乙锭(AO/EB)荧光染色法检测细胞凋亡与坏死.结果表明:与对照组比较,0.625,1.250和2.500 mg/mL佛手水煎剂处理后,RAW 264.7癌细胞固缩,细胞核染色质凝聚、片断化,有凋亡小体出现,表现出典型的细胞凋亡特征;5.000 mg/mL佛手水煎剂处理后,RAW 264.7癌细胞肿胀,细胞膜破裂,呈现坏死症状,RAW 264.7癌细胞的增殖明显受到抑制(P＜0.01),半数抑制浓度(IC50)为2.073 mg/mL.说明佛手水煎剂能诱导癌细胞凋亡,抑制癌细胞增殖.     

Conditional telomerase induction causes proliferation of hair follicle stem cells

TERT, the protein component of telomerase, serves to maintain telomere function through the de novo addition of telomere repeats to chromosome ends, and is reactivated in 90% of human cancers. In normal tissues, TERT is expressed in stem cells and in progenitor cells, but its role in these compartments is not fully understood. Here we show that conditional transgenic induction of TERT in mouse skin epithelium causes a rapid transition from telogen (the resting phase of the hair follicle cycle) to anagen (the active phase), thereby facilitating robust hair growth. TERT overexpression promotes this developmental transition by causing proliferation of quiescent, multipotent stem cells in the hair follicle bulge region. This new function for TERT does not require the telomerase RNA component, which encodes the template for telomere addition, and therefore operates through a mechanism independent of its activity in synthesizing telomere repeats. These data indicate that, in addition to its established role in extending telomeres, TERT can promote proliferation of resting stem cells through a non-canonical pathway.     

Hedgehog/Wnt feedback supports regenerative proliferation of epithelial stem cells in bladder

Epithelial integrity in metazoan organs is maintained through the regulated proliferation and differentiation of organ-specific stem and progenitor cells. Although the epithelia of organs such as the intestine regenerate constantly and thus remain continuously proliferative, other organs, such as the mammalian urinary bladder, shift from near-quiescence to a highly proliferative state in response to epithelial injury. The cellular and molecular mechanisms underlying this injury-induced mode of regenerative response are poorly defined. Here we show in mice that the proliferative response to bacterial infection or chemical injury within the bladder is regulated by signal feedback between basal cells of the urothelium and the stromal cells that underlie them. We demonstrate that these basal cells include stem cells capable of regenerating all cell types within the urothelium, and are marked by expression of the secreted protein signal Sonic hedgehog (Shh). On injury, Shh expression in these basal cells increases and elicits increased stromal expression of Wnt protein signals, which in turn stimulate the proliferation of both urothelial and stromal cells. The heightened activity of this signal feedback circuit and the associated increase in cell proliferation appear to be required for restoration of urothelial function and, in the case of bacterial injury, may help clear and prevent further spread of infection. Our findings provide a conceptual framework for injury-induced epithelial regeneration in endodermal organs, and may provide a basis for understanding the roles of signalling pathways in cancer growth and metastasis.     

Increased proliferation of B cells and auto-immunity in mice lacking protein kinase Cdelta

Protein kinase C (PKC), which comprises 11 closely related isoforms, has been implicated in a wide variety of cellular processes, such as growth, differentiation, secretion, apoptosis and tumour development. Among the PKC isotypes, PKC-delta is unique in that its overexpression results in inhibition of cell growth. Here we show that mice that lack PKC-delta exhibit expansion of the B-lymphocyte population with the formation of numerous germinal centres in the absence of stimulation. The rate of proliferation in response to stimulation was greater for B cells from PKC-delta-deficient mice than for those from wild-type mice. Adoptive transfer experiments suggested that the hyperproliferation phenotype is B-cell autonomous. Production of interleukin-6 was markedly increased in B cells of PKC-delta-null mice as a result of an increase in the DNA-binding activity of NF-IL6. Furthermore, the PKC-delta-deficient mice contain circulating autoreactive antibodies and display immune-complex-type glomerulonephritis, as well as lymphocyte infiltration in many organs. These results suggest that PKC-delta has an indispensable function in negative regulation of B-cell proliferation, and is particularly important for the establishment of B-cell tolerance.