Stem cells, cancer, and cancer stem cells.

Stem cell biology has come of age. Unequivocal proof that stem cells exist in the haematopoietic system has given way to the prospective isolation of several tissue-specific stem and progenitor cells, the initial delineation of their properties and expressed genetic programmes, and the beginnings of their utility in regenerative medicine. Perhaps the most important and useful property of stem cells is that of self-renewal. Through this property, striking parallels can be found between stem cells and cancer cells: tumours may often originate from the transformation of normal stem cells, similar signalling pathways may regulate self-renewal in stem cells and cancer cells, and cancer cells may include 'cancer stem cells' - rare cells with indefinite potential for self-renewal that drive tumorigenesis.     

Stem cells, ageing and the quest for immortality

Adult stem cells reside in most mammalian tissues, but the extent to which they contribute to normal homeostasis and repair varies widely. There is an overall decline in tissue regenerative potential with age, and the question arises as to whether this is due to the intrinsic ageing of stem cells or, rather, to the impairment of stem-cell function in the aged tissue environment. Unravelling these distinct contributions to the aged phenotype will be critical to the success of any therapeutic application of stem cells in the emerging field of regenerative medicine with respect to tissue injury, degenerative diseases or normal functional declines that accompany ageing.     

Growth regulation of a cellular tumour antigen, p53, in nontransformed cells

Many transformed cells in culture have been found to express elevated levels of a cellular tumour antigen, termed p53. This protein has also been implicated in the regulation of cellular growth. For these reasons experiments were designed to examine the expression of p53 as quiescent cultures of nontransformed 3T3 fibroblasts were stimulated to reenter the cell cycle. Synchronous populations of cells were obtained by releasing a culture from density-dependent inhibition of growth with the addition of fresh serum. Steady-state levels of p53 protein and mRNA were measured as a function of time after addition of serum to quiescent cultures and the rate of synthesis of p53 protein was analysed at a number of time points. The results, reported here, demonstrate an increase in the synthesis and steady-state levels of p53 protein and mRNA prior to DNA synthesis in late G1, and suggest a role for p53 in the progression of cells from a growth-arrested state to an actively dividing state.     

Fabrication, nanostructures and electronic properties of nanodiamond-based solar cells

Nanodiamond-based solar cells were fa bricated and the photovoltaic properties were investigated. Fullerene (C60) and fullerenol (C60(OH)10-12) were used as n-type semiconductors, and diamond nan oparticles and metal phthalocyanine derivative were used as p-type semiconductors. The nanostructures of the solar cel ls were investigated by transmission electron microscopy and X-ray diffracometry, and the electronic property was discussed.     

A microenvironment,rather than chemical,initiates the cardiomyogenic differentiation of marrow stromal cells

To investigate the potential of cardiomyogenic differentiation of rat hone marrow stromal cells (MSCs), they were exposed to 5-azacytidine treatments (single/repeat) at varying concentrations (3, 5, 10μmol/L) and the fates of the cells were analyzed by immunocytochemistry, Western blot and the reporter gene of enhanced cyan fluorescent protein (ECFP) under the control of ventricular myosin light chain 2 (MLC2v) promoter. MSCs were also cocultured with cardiomyocytes for periods up to 16 days, and the expression of cardiac myosin heavy chain(MHC) and troponin Ⅰ (Tn I) proteins was analyzed. After the induction with 5-azacytidine, neither spontaneously beating ceils nor myotubes were found; MHC and Tn I proteins were also undetectable and no ECFP-positive MSCs were detected. But when cocultured with cardiomyocytes, spontaneously contracting MSCs were observed and cardiac specific proteins could be detected. The results proved that the novel effects of 5-azacytldine on the cardiomyogenic differentiation of MSCs should be questioned and a direct intercellular communication with cardiomyocytes is necessary for MSCs to differentiate into cardiomyocytes.     

Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes

Recent studies have suggested that bone marrow cells possess a broad differentiation potential, being able to form new liver cells, cardiomyocytes and neurons. Several groups have attributed this apparent plasticity to 'transdifferentiation'. Others, however, have suggested that cell fusion could explain these results. Using a simple method based on Cre/lox recombination to detect cell fusion events, we demonstrate that bone-marrow-derived cells (BMDCs) fuse spontaneously with neural progenitors in vitro. Furthermore, bone marrow transplantation demonstrates that BMDCs fuse in vivo with hepatocytes in liver, Purkinje neurons in the brain and cardiac muscle in the heart, resulting in the formation of multinucleated cells. No evidence of transdifferentiation without fusion was observed in these tissues. These observations provide the first in vivo evidence for cell fusion of BMDCs with neurons and cardiomyocytes, raising the possibility that cell fusion may contribute to the development or maintenance of these key cell types.     

Weighing of biomolecules, single cells and single nanoparticles in fluid

Nanomechanical resonators enable the measurement of mass with extraordinary sensitivity. Previously, samples as light as 7 zeptograms (1 zg = 10(-21) g) have been weighed in vacuum, and proton-level resolution seems to be within reach. Resolving small mass changes requires the resonator to be light and to ring at a very pure tone-that is, with a high quality factor. In solution, viscosity severely degrades both of these characteristics, thus preventing many applications in nanotechnology and the life sciences where fluid is required. Although the resonant structure can be designed to minimize viscous loss, resolution is still substantially degraded when compared to measurements made in air or vacuum. An entirely different approach eliminates viscous damping by placing the solution inside a hollow resonator that is surrounded by vacuum. Here we demonstrate that suspended microchannel resonators can weigh single nanoparticles, single bacterial cells and sub-monolayers of adsorbed proteins in water with sub-femtogram resolution (1 Hz bandwidth). Central to these results is our observation that viscous loss due to the fluid is negligible compared to the intrinsic damping of our silicon crystal resonator. The combination of the low resonator mass (100 ng) and high quality factor (15,000) enables an improvement in mass resolution of six orders of magnitude over a high-end commercial quartz crystal microbalance. This gives access to intriguing applications, such as mass-based flow cytometry, the direct detection of pathogens, or the non-optical sizing and mass density measurement of colloidal particles.     

3,7-二硝基-二苯基溴盐诱导K562细胞株的凋亡

采用苔盼蓝染色法、MTT比色法研究了环状溴代鎓盐cBr对人慢性粒细胞白血病细胞株K562增殖的影响;并用同位素示踪技术(3H-TdR)研究了cBr对K562细胞DNA的损伤作用;从形态学(荧光显微镜、电镜观察)和生化特性(流式细胞术、3H-TdR掺入法分析)研究了cBr抑制K562细胞增殖作用的机制.结果表明:在体外cBr显著抑制K562细胞增殖,并对DNA造成损伤,提示cBr的作用是通过诱导细胞凋亡而进行的,是一种新型的免疫增强剂.     

间质干细胞来源、鉴定、可塑性和应用前景(综述)

间质干细胞(MSCs)存在于骨髓、肌肉、骨、软骨等部位，可分化为中胚层的细胞，如成骨细胞、成软骨细胞、成脂肪细胞等，亦可分化为内胚层、外胚层的细胞，如神经细胞、肝脏细胞、肾脏细胞等，同时由于来源广、易于分离扩增和基因转染，在临床、科研上有很大的潜在应用价值。     

马钱子苷对神经干细胞的增殖、存活和分化的调节作用

为探讨不同浓度的马钱子苷对神经干细胞的增殖、存活和分化的调节作用及其相关分子机制,本实验从成年小鼠大脑中分离培养了神经干细胞,用不同浓度的马钱子苷进行干预,观察马钱子苷对神经干细胞增殖、存活和分化的影响。结果显示：成年小鼠神经干细胞在含有中、高浓度马钱子苷的增殖培养基中培养5 d和7 d后,神经球数量和直径与对照相比显著增加（P<0.05）;中、高剂量的马钱子苷能够促进神经干细胞的有丝分裂,低剂量马钱子苷处理显著促进神经干细胞发生分化（P<0.01）,并增加神经元和星形胶质细胞的数量及比例（P<0.01）;中、高剂量马钱子苷抑制神经干细胞分化（P<0.05）,高剂量的马钱子苷使得神经元的数量减少（P<0.05）。研究结果表明,高浓度的马钱子苷能够促进神经干细胞存活,并通过促进神经干细胞有丝分裂来提高其增殖能力;低浓度的马钱子苷促进神经干细胞分化,有利于神经再生和少突胶质细胞再生。研究结果为神经干细胞治疗中枢神经系统疾病的研究奠定了理论和实验基础。     

Distribution of CRH-and ACTH-like immunoreactive cells in amphioxus,Branchiostoma belcheri

Immunocytochemical studies on the nervous system,Hatschek's pit,digestive tract and gonads tissues of an amphioxus(Branchiostoma belcheri)were performed using polyclonal antibodies against human corticotrophin-releasing hormone(CRH)and human adrenocorticotropin(ACTH).The results showed that many CRH-like immunoreactive neurons were distributed specifically on the dorsal side and ventral side of brain vesicle,while a few CRH-like neurons and their fibers in spinal cord.At the same time,the epithelial cells in the basic region of Hatschek's pit were shown immunopositive to CRH antibody.In gonads(ovary and testis),CRH-immunopositive substance was localized in the cytoplasm near oocyte nucleus and in early spermatogenic cells.ACTH-like immunoreactivities were observed specially in the neurons and their protrusions localized on the ventral side of the brain vesicle and in spinal cord,and also in epithelial cells of Hatschek's pit,enteric neurons of digestive tract,oocytes in ovary and in early spermatogenic cells as well.It was found for the first time that CRH-like neurons existed in the middle region of brain vesicle(corresponding to the hypothalamus of vertebrates)and ACTH-like immunopositive cells existed in Hatschek's pit,implying that a control mechanism between brain vesicle and Hatschek's pit maybe had been already built in amphioxus as that in vertebrates.The present study will provide new morphological evidence for the origin and evolution of ACTH.In addition,the immunoreactivities of CRH and ACTH in the digestive tract and gonads suggested other physiological function of CRH and ACTH in amphioxus.     

循环肿瘤细胞分离富集与分析检测技术的研究进展

癌症死亡患者中有90%由癌症转移引起,研究表明:患者的外周血、胸腔液等体液中的循环肿瘤细胞(CTC)与癌症转移及肿瘤结节转移(TNM)分期密切相关.因此,CTC检测在实体肿瘤前期诊断、预后及疗效评估等方面扮演着举足轻重的角色.该综述基于CTC区别于正常细胞的物理学、电学、生物学特征,总结了目前CTC分离富集及分析检测技术的研究进展,并就国内外CTC检测面临的挑战进行了讨论,对其未来发展趋势进行了展望.     

Distribution of CRH- and ACTH-like immunoreactive cells in amphioxus, Branchiostoma belcheri

Immunocytochemical studies on the nervous system, Hatschek’s pit，digestive tract and gonads tissues of an amphioxus (Branchiostoma belcheri) were performed using polyclonal antibodies against human corticotrophin-releasing hormone (CRH) and human adrenocorticotropin (ACTH). The results showed that many CRH-like immunoreactive neurons were distributed specifically on the dorsal side and ventral side of brain vesicle, while a few CRH-like neurons and their fibers in spinal cord. At the same time, the epithelial cells in the basic region of Hatschek’s pit were shown immunopositive to CRH antibody. In gonads (ovary and testis), CRH-immunopositive substance was localized in the cytoplasm near oocyte nucleus and in early spermatogenic cells. ACTH-like immunoreactivities were observed specially in the neurons and their protrusions localized on the ventral side of the brain vesicle and in spinal cord, and also in epithelial cells of Hatschek’s pit, enteric neurons of digestive tract, oocytes in ovary and in early spermatogenic cells as well. It was found for the first time that CRH-like neurons existed in the middle region of brain vesicle (corresponding to the hypothalamus of vertebrates) and ACTH-like immunopositive cells existed in Hatschek’s pit, implying that a control mechanism between brain vesicle and Hatschek’s pit maybe had been already built in amphioxus as that in vertebrates. The present study will provide new morphological evidence for the origin and evolution of ACTH. In addition, the immunoreactivities of CRH and ACTH in the digestive tract and gonads suggested other physiological function of CRH and ACTH in amphioxus.     

JWA,a novel microtubule-associated protein,regulates homeostasis of intracellular amino acids in PC12 cells

Microtubules are long, stiff polymers that extendthroughout the cytoplasm and govern the location of membrane-bounded organelles and other cell components.The building block of a microtubule is the tubulin subunit, a heterodimer of a- and b-tubulin. Both of these 50 kDmonomers are found in all eukaryotes, and their se-quences are highly conserved. The bulk of cellular tubulin is cytoplasmic, but a significant fraction is embedded in, or firmly associated with the plasma membrane and other memb…     

JWA, a novel microtubule－associated protein, regulates homeostasis of intracellular amino acids in PC12 cells

Our previous studies demonstrated that JWA,a novel retinoic acids responsive and cytoskeleton related gene, is associated with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a novel kind of microtubnle-associated proteins (MAPs) and functiona llylink to microtubule, we first successfully identified JWA from the physically purified MAPs complex of rat braint issues. The results of co-immunoprecipitation, gene transfection and immunofluorescence microscopy assays from HBE and NIH3T3 cells provide strong evidence for a linkage between JWA and β-tubulin. In general, JWA is stably binding to β-tubulin whenever microtubule is polymerized or not,and it may be critical to the mitosis process. In addition, by use of the antisense oligonucleotides technique, we also showed that JWA is a negative modulator on intracellular amino acids in PC12 cells. Further analysis indicated that JWA selectively regulates both taurine, an inhibitory aminoacid, and glutamate, an excitatory amino acid. In conclusion,JWA is not only structurally associated, but also a novel functional MAP.