利用内源性干细胞原位再生晶状体治疗婴幼儿白内障

 利用内源性干细胞进行组织器官的修复和再生是再生医学研究的最终目标。婴幼儿白内障是幼儿致盲性眼病的首要病因,目前尚无有效治疗手段。本研究组从哺乳动物内成功分离并获得了晶状体上皮干细胞,证明Pax6和Bmi1是维持其分化和自我更新的关键因子,并以保留内源性干细胞为目标,设计了全新的白内障术式。相较于传统术式,新术式最大程度地保留了内源性干细胞、基底膜和微环境,在新西兰兔、食蟹猴和先天性白内障患儿内实现了功能性晶状体的再生。研究结果为白内障提供了全新的治疗策略并为组织再生及内源性干细胞的应用提供了全新的范例。     

莪术油对人胃腺癌SGC-7901细胞形态的影响

研究莪术油(Zedoary Turmeric Oil,ZTO)对体外培养的人胃腺癌SGC-7901细胞形态变化的影响。不同浓度的ZTO作用人胃腺癌SGC-7901细胞24 h后,通过普通光学显微镜、荧光显微镜、透射电镜观察细胞形态变化情况。光学显微镜和荧光显微镜下可见未加药物组细胞呈梭形,胞浆丰富,核质均匀,核仁呈现黄色荧光,加药组细胞体积变小,胞浆浓缩、胞膜有起泡脱落现象,胞膜崩解,部分细胞核破碎或消失,细胞核呈现绿色或亮黄色荧光。透射电镜下可见未加药组细胞呈圆形,细胞核较大,胞浆较多,细胞器丰富,加药组细胞积明显变小,核固缩,微绒毛消失,部分坏死细胞的胞核溶解,细胞器结构崩解消失。ZTO作用人胃腺癌SGC-7901细胞24 h能致见未加药组细胞胞浆较多,细胞器丰富,加药组细胞体积明显变小,坏死细胞的胞使细胞发生明显的形态改变,且具有剂量依赖性。     

Endonuclease G is an apoptotic DNase when released from mitochondria.

Nucleosomal fragmentation of DNA is a hallmark of apoptosis (programmed cell death), and results from the activation of nucleases in cells undergoing apoptosis. One such nuclease, DNA fragmentation factor (DFF, a caspase-activated deoxyribonuclease (CAD) and its inhibitor (ICAD)), is capable of inducing DNA fragmentation and chromatin condensation after cleavage by caspase-3 (refs 2,3,4). However, although transgenic mice lacking DFF45 or its caspase cleavage site have significantly reduced DNA fragmentation, these mice still show residual DNA fragmentation and are phenotypically normal. Here we report the identification and characterization of another nuclease that is specifically activated by apoptotic stimuli and is able to induce nucleosomal fragmentation of DNA in fibroblast cells from embryonic mice lacking DFF. This nuclease is endonuclease G (endoG), a mitochondrion-specific nuclease that translocates to the nucleus during apoptosis. Once released from mitochondria, endoG cleaves chromatin DNA into nucleosomal fragments independently of caspases. Therefore, endoG represents a caspase-independent apoptotic pathway initiated from the mitochondria.     

酿酒酵母超微结构观察方法的研究

本文用常规电镜技术对不同细胞周期的酿酒酵母ＳａｃｃｈａｒｏｍｙｃｅｓＣｅｒｅｖｉｓｉａｅ菌体细胞和原生质体进行超微结构的对比观察，结果发现，用减数分裂时期的原生质体进行超微结构的观察，效果最为理想。包括细胞核在内的许多细胞器都可观察到。纺锤极体，微管，染色质，染色体，核仁等核内结构也都十分清晰。     

Mechanisms of plant somatic embryogenesis

Based on our experimental results and the related research reports, this review addresses some important issues about plant somatic embryogenesis. Once plant somatic cells develop into embryonic cells, the organelles of the cells increase, and their function is active. In the early stage of embryogenic cells, the reaction products of AT-Pase are mainly deposited on the plasma and vacuole's membranes, then the activity of ATPase can be found in the cytoplasm, vacuole and nucleus in the late stage of embryogenic cells. ATPase-catalyzed reaction occurs actively in the thickened wall of embryogenic cells. Endogenous hormone is a critical factor affecting somatic embryogenesis. Superox-ide dismutase (SOD) activity, which is closely related to embryogenic cell differentiation, is much higher in differentiated embryogenic cells than in undifferentiated cells, indicating that embryogenic cells have stronger function of antioxida-tion. In addition, the lower level H2O2 can promote embryogenic cell differentiation. The programmed cell death (PCD) exists in the process of embryogenic cell differentiation and development, and the active oxygen species plays an important role in inducing of plant PCD.     

润楠雄配子体发育的超微结构研究 Ⅲ.营养细胞和生殖细胞的形成

对润楠(Machilus pingii Cheng ex Yang)花粉生殖细胞和营养细胞的形成进行了观察.结果如下:(1)随着小孢子核向(?)方向迁移,胞质的线粒体和前质体显著增多;由于大液泡的存在,细胞器偏向核的一侧分布.(2)小孢子有丝分裂末期,胞质中出现较多的高尔基体和微管.(3)生殖细胞(?)抵达花粉内壁后,生殖细胞和营养细胞的胞质中出现大量与壁平行排列的粗面内质网.随着生殖细胞壁的加厚, 生殖细胞的局部与营养细胞核相贴近,贴近区域的生殖细胞质中出现交织成网的内质网.(4)游离的生殖细胞具,胞质中仅有少量细胞器;营养细(?)中储存有大量的淀粉粒.从观察结果推测:影响润楠生殖细胞在花粉内进一步分裂的因素可能有三种:一是生殖细胞壁留存的时间;二是营养细胞和生殖细胞本身的代谢状况;三是营养细胞核对生殖细胞的作用.     

Chronic polyarthritis caused by mammalian DNA that escapes from degradation in macrophages

A large amount of chromosomal DNA is degraded during programmed cell death and definitive erythropoiesis. DNase II is an enzyme that digests the chromosomal DNA of apoptotic cells and nuclei expelled from erythroid precursor cells after macrophages have engulfed them. Here we show that DNase II-/-IFN-IR-/- mice and mice with an induced deletion of the DNase II gene develop a chronic polyarthritis resembling human rheumatoid arthritis. A set of cytokine genes was strongly activated in the affected joints of these mice, and their serum contained high levels of anti-cyclic citrullinated peptide antibody, rheumatoid factor and matrix metalloproteinase-3. Early in the pathogenesis, expression of the gene encoding tumour necrosis factor (TNF)-alpha was upregulated in the bone marrow, and administration of anti-TNF-alpha antibody prevented the development of arthritis. These results indicate that if macrophages cannot degrade mammalian DNA from erythroid precursors and apoptotic cells, they produce TNF-alpha, which activates synovial cells to produce various cytokines, leading to the development of chronic polyarthritis.     

Homologies between gap junction proteins in lens, heart and liver

The cells in the mammalian lens are electrically and metabolically coupled with each other by a network of gap junctions. These are clusters of transmembrane channels by which the fibre cells situated deeper in the lens communicate through the epithelium with the aqueous humour, the source of nutrients for the lens. Hence gap junctions are important for lens transparency. The gap junction proteins in the mammalian lens have not yet been identified with certainty. A putative fibre gap junction protein of relative molecular mass 26,000 (26K) is not related to those from other tissues, such as the liver 28K junction component. Another lens membrane protein with Mr 70K (MP70) has also been localized in the lens fibre gap junctions. Here we demonstrate by amino-terminal sequence analysis that MP70 and its in vivo-processed form, MP38 (ref. 8), belong to a wider family of gap junction proteins. With this new data on the lens, homologies between gap junction proteins now extend to organs derived from all three embryonal layers, endoderm (liver), mesoderm (heart) and ectoderm (lens).     

水稻精细胞的超微结构

在稻成熟花粉粒中，精细胞被不连续的电子透明的周质空间所包围，纵切面上显示的精细胞显著的由细胞质延伸形成的尾状结构，主要为长圆形的细胞核所占据的一端为头部，精细胞中具有一般的细胞器，其中线粒体主要位于头部，微丝成束与细胞纵轴平行排列，观察表明，在水稻成熟花粉中具有雄性生殖单位，两个精细胞以尾部紧密连接，其中一个精细胞深入到营养细胞核中。     

IDH1(R132H) mutation increases murine haematopoietic progenitors and alters epigenetics

Mutations in the IDH1 and IDH2 genes encoding isocitrate dehydrogenases are frequently found in human glioblastomas and cytogenetically normal acute myeloid leukaemias (AML). These alterations are gain-of-function mutations in that they drive the synthesis of the ‘oncometabolite’ R-2-hydroxyglutarate (2HG). It remains unclear how IDH1 and IDH2 mutations modify myeloid cell development and promote leukaemogenesis. Here we report the characterization of conditional knock-in (KI) mice in which the most common IDH1 mutation, IDH1(R132H), is inserted into the endogenous murine Idh1 locus and is expressed in all haematopoietic cells (Vav-KI mice) or specifically in cells of the myeloid lineage (LysM-KI mice). These mutants show increased numbers of early haematopoietic progenitors and develop splenomegaly and anaemia with extramedullary haematopoiesis, suggesting a dysfunctional bone marrow niche. Furthermore, LysM-KI cells have hypermethylated histones and changes to DNA methylation similar to those observed in human IDH1- or IDH2-mutant AML. To our knowledge, our study is the first to describe the generation and characterization of conditional IDH1(R132H)-KI mice, and also the first report to demonstrate the induction of a leukaemic DNA methylation signature in a mouse model. Our report thus sheds light on the mechanistic links between IDH1 mutation and human AML.     

Monoclonal mice generated by nuclear transfer from mature B and T donor cells

Cloning from somatic cells is inefficient, with most clones dying during gestation. Cloning from embryonic stem (ES) cells is much more effective, suggesting that the nucleus of an embryonic cell is easier to reprogram. It is thus possible that most surviving clones are, in fact, derived from the nuclei of rare somatic stem cells present in adult tissues, rather than from the nuclei of differentiated cells, as has been assumed. Here we report the generation of monoclonal mice by nuclear transfer from mature lymphocytes. In a modified two-step cloning procedure, we established ES cells from cloned blastocysts and injected them into tetraploid blastocysts to generate mice. In this approach, the embryo is derived from the ES cells and the extra-embryonic tissues from the tetraploid host. Animals cloned from a B-cell nucleus were viable and carried fully rearranged immunoglobulin alleles in all tissues. Similarly, a mouse cloned from a T-cell nucleus carried rearranged T-cell-receptor genes in all tissues. This is an unequivocal demonstration that a terminally differentiated cell can be reprogrammed to produce an adult cloned animal.     

Degree of methylation of transgenes is dependent on gamete of origin

Data derived from both pronuclear transplantation experiments and classical genetic experiments indicate that the maternal and paternal genetic contributions to the mammalian zygote nucleus do not function equivalently during subsequent development. These observations have been interpreted as resulting from differential 'genome imprinting' during male and female gametogenesis. The molecular mechanism responsible for genome imprinting is unknown, but data gathered to date require that the mechanism fulfill at least four criteria: (1) the imprint must be physically linked to the pronucleus; (2) the imprint must persist through DNA replication and cell division; (3) the mechanism must be capable of affecting gene expression; and (4) the mechanism must be capable of switching the identity of the imprint from one sex to the other in successive generations. One molecular mechanism which could satisfy the first three criteria is differential DNA methylation during gametogenesis itself, or before formation of the zygote nucleus during embryogenesis. We present data indicating that the methylation patterns of exogenous DNA sequences in transgenic mice can be changed by switching their gamete of origin in successive generations. These data suggest that DNA methylation can also satisfy the fourth criterion for an imprinting mechanism.     

Myxococcus xanthus spore coat protein S may have a similar structure to vertebrate lens beta gamma-crystallins

The Gram-negative bacterium Myxococcus xanthus has a complex life cycle during which large amounts of a protein of relative molecular mass (Mr) 19,000, known as protein S, are assembled into a spore surface coat by a process that specifically requires calcium ions. The gene for protein S has been cloned and the DNA sequence shows that the gene product is composed of four internally repeated homologous sequences, each 40 amino acids long. Although protein S resembles calmodulin both in its internally duplicated structure and its ability to bind calcium, it apparently has a beta-sheet secondary structure rather than the helix-loop-helix motifs that characterize the calmodulin family. We now show that protein S has a striking homology with the beta- and gamma-crystallins of the vertebrate eye lens which are beta-sheet proteins with internally duplicated structures. This implies that the beta- and gamma-crystallins evolved from already existing proteins, whose ancestors occurred in the prokaryotes. The biological function of protein S, as a closely packed, stable protein in a relatively dehydrated environment, has implications for the functions of crystallins, which are found closely packed in the lens fibre cells, where their stability is essential for maintenance of transparency.     

花背蟾蜍变态前后眼各部位Con A受体的荧光显微镜观察

本文以花背蟾蜍变态前后的眼为材料,运用 Con A-FITC 在荧光显微镜下观察了眼变态前后 Con A 受体的变化.结果表明:变态前內处角膜愈合处的 Con A 受体较多;晶体上皮的外表面比内表面 Con A 受体多;视网膜各层扬构的 Con A 受体分布差异较大.变态后角膜上皮的细胞膜,角膜基质和角膜内皮中均有 Con A 受体分布;晶状体仅晶体上皮有 Con A受体分布;视网膜中内界膜,内网层,外网层,外界膜,视杆视锥细胞层和色素上皮层的内外表面 Con A 受体较多:节细胞层,内核层和外核层仅细胞表面有少量的 Con A 受体分布.推测眼在变态前后 Con A 受体数量和分布上的变化与眼各部位细胞的分化,结构的组建和视觉功能的完善有关.     

Mitochondrial endonuclease G is important for apoptosis in C. elegans.

Programmed cell death (apoptosis) is a tightly regulated process of cell disassembly in which dying cells and their nuclei shrink and fragment and the chromosomal DNA is degraded into internucleosomal repeats. Here we report the characterization of the cps-6 gene, which appears to function downstream of, or in parallel to, the cell-death protease CED-3 of Caenorhabditis elegans in the DNA degradation process during apoptosis. cps-6 encodes a homologue of human mitochondrial endonuclease G, and its protein product similarly localizes to mitochondria in C. elegans. Reduction of cps-6 activity caused by a genetic mutation or RNA-mediated interference (RNAi) affects normal DNA degradation, as revealed by increased staining in a TUNEL assay, and results in delayed appearance of cell corpses during development in C. elegans. This observation provides in vivo evidence that the DNA degradation process is important for proper progression of apoptosis. CPS-6 is the first mitochondrial protein identified to be involved in programmed cell death in C. elegans, underscoring the conserved and important role of mitochondria in the execution of apoptosis.     

Maintenance of optical quality during crystalline lens growth

Vertebrate lenses grow throughout life by the division of cells at the lens surface. The fibre cells thus produced are gradually covered by newer tissue, giving a layered structure. During growth, the lens must remain transparent and retain its refractile properties. The severity of these constraints is perhaps most evident in teleost fish which have a spherical lens that may increase in volume by a thousandfold during the first year of life. The dioptric power of the teleost fish eye is vested entirely in this spherical lens, as water, the cornea and the intraocular vitreous humour have almost identical refractive indices. Spherical lenses of uniform refractive index produce poor images because rays entering at different distances from the optic axis are focused at different distances from the lens. Teleost fish do not suffer from this imperfection and it has long been presumed that this is because there exists a refractive index gradient having a high value in the centre and decreasing continuously and symmetrically with radius in all directions. Here we demonstrate in the African cichlid fish, Haplochromis burtoni, that a refractive index gradient does exist, although its form is significantly different from that previously postulated.     

Apoptosis in suspension culture of tobacco cells induced by heat shock

The induction of apoptosis in suspension culture of tobacco cells by heat shock is reported for the first time. Heat treatment (48℃ for 4 h) of tobacco cells led to the appearance of typical hallmarks of apoptosis. It was demonstrated by DNA laddering analysis that the cells treated with heat shock at 48℃ for 4 h had a serious degradation of nuclear DNA into multi-nu-cleosomal sizes, suggesting that heat shock activated endogenous nuclease which led to DNA cleavage at the linkage sites between the nucleosomes, but ladders were very faint for DNA from 2 and 9 h heat-treated cells. The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) detection also showed that most of these treated cells (48℃ for 4 h) displayed positive reactions, indicating a serious DNA 3'-OH cleavage in their nuclei. Moreover, some other cytological changes in apoptotic cells, such as cell shrinkage, chromatin aggregation, nucleus collapse, have also been observed by 4', 6'-diamidino-2-phenyl-indole (DAPI) staining.     

Regulated degradation of a class V myosin receptor directs movement of the yeast vacuole

Normal cellular function requires that organelles be positioned in specific locations. The direction in which molecular motors move organelles is based in part on the polarity of microtubules and actin filaments. However, this alone does not determine the intracellular destination of organelles. For example, the yeast class V myosin, Myo2p, moves several organelles to distinct locations during the cell cycle. Thus the movement of each type of Myo2p cargo must be regulated uniquely. Here we report a regulatory mechanism that specifically provides directionality to vacuole movement. The vacuole-specific Myo2p receptor, Vac17p, has a key function in this process. Vac17p binds simultaneously to Myo2p and to Vac8p, a vacuolar membrane protein. The transport complex, Myo2p-Vac17p-Vac8p, moves the vacuole to the bud, and is then disrupted through the degradation of Vac17p. The vacuole is ultimately deposited near the centre of the bud. Removal of a PEST sequence (a potential signal for rapid protein degradation) within Vac17p causes its stabilization and the subsequent 'backward' movement of vacuoles, which mis-targets them to the neck between the mother cell and the bud. Thus the regulated disruption of this transport complex places the vacuole in its proper location. This may be a general mechanism whereby organelles are deposited at their terminal destination.