成纤维细胞和纤维肉瘤细胞的中等纤维构型特点及其意义

对体外培养的成纤维细胞，高分化纤维肉瘤细胞、低转移性及高转移性纤维肉瘤细胞的中等纤维的构型进行了研究。结果显示：中等纤维在恶变细胞中，其构型经发生紊乱，并随细胞恶变程度的增加，其紊乱的程度也相应增加。说明中等纤维构型变化紊乱是恶变细胞的异型性表现之一。转移率高的瘤细胞，这种异型性更大。     

钙调素拮抗剂TFP对Hela细胞微丝及波形纤维蛋…

利用钙调素拮抗剂三氟拉素（ＴＦＰ）及微管的不同聚合状态对Ｈｅｌａ细胞微细及波形纤维白分布的影响进行了。Ｈｅｌａ细胞微丝明显解聚，而波形纤维蛋白则密集分布在细胞一侧。细胞经ＴＦＰ处理２ｄ后，可见微丝分布的恢复，而波形纤维蛋白分布则变得分散，并向质膜延伸，但对ＴＦＰ处理２ｄ后的细胞用低温（２－４℃）处理，使微管解聚，或以紫杉酚处理２４ｈ，使微管高度聚集以破坏其微客网络系统时，微丝则随之发生明显的解聚现     

超低频磁场对鼠肉瘤细胞核参数的影响

用超低频脉磁场处理鼠Ｓ－１８０肉瘤细胞并进行光镜观察；应用计算机图像分析技术对细胞核参数进行分析。结果表明：磁疗后肉瘤细胞形态变化显著，核平均面积和周长减小，圆形度差异不明显，ＤＮＡ倍性降低，认为该磁场抑制Ｓ－１８０肉瘤细胞的生长，并对其机理做了初步探讨。     

超低频磁场对鼠肉瘤细胞核参数的影响

用超低频脉冲磁场处理鼠Ｓ－１８０肉瘤细胞并进行光镜观察；应用计算机图像分析技术对细胞核参数进行分析．结果表明：磁疗后肉瘤细胞形态变化显著，核平均面积和周长减小，圆形度差异不明显，ＤＮＡ倍性降低．认为该磁场抑制了Ｓ－１８０肉瘤细胞的生长，并对其机理做了初步探讨．     

影响人软组织肉瘤异种移植成功的某些相关因素

在裸小鼠体内移植了１４例肉瘤，成功５例（２例恶性纤维组织细胞瘤，２例横纹肌肉瘤，１例恶性外周细胞血管瘤），失败９例，接种成功率为３６％。分成移植成功群（ｓｕｃｃｅｅｄｅｄｉｎｔｒａｎｓｐｌａｎｔ，Ｓ群）和移植失败群（ｆａｉｌｅｄｔｏｔｒａｎｓｐｌａｎｔ，Ｆ群），对１４例被移植肉瘤的临床状况、组织病理学、免疫特性、ＤＮＡ含量、增殖因子和癌基因蛋白等进行了比较研究。Ｓ群中４例ＡＪＣ分期为Ⅲ或Ⅳ期，５例都是复发病例。Ｓ群的恶性程度明显高于Ｆ群，其中４例为高度恶性肉瘤，表现为细胞密度高，核异形性大，核分裂像显著增多。结果表明与肉瘤异种移植成功有关的某些可能因素为：组织来源，恶性程度高低，间质成份多寡，Ｓ期细胞百分数等。免疫学特性对肉瘤异种移植成功无明显影响。增殖因子Ｋｉ－６７和ＰＣＮＡ在Ｓ群中的表达明显高于Ｆ群，Ｓ群标本通常呈现较强的Ｋｉ－６７和ＰＣＮＡ染色，而Ｆ群通常显示较弱的染色，表明这两者是判定肉瘤生长速率和进展的较理想的指标。癌基因蛋白Ｐ５３在Ｓ群中的阳性表达率（６０％）明显高于下群（１８％）．进一步证明Ｐ５３在肿瘤进展中的重要作用。余癌某因蛋白在两群间的表达率无显著差别。     

人肝癌细胞骨架网络系统的初步研究

利用ＴｒｉｔｏｎＸ－１００及其联合（ＮＨ４）２ＳＯ４的抽提技术与ＣｏｏｍａｓｉｅｂｌｕｅＲ２５０染色、免疫酶标技术相结合，体外实验比较研究人肝癌细胞系（ＨｅｐＧ２）细胞骨架网络的分布构像及其中间纤维蛋白构型。结果：人肝癌细胞系（ＨｅｐＧ２）显示其细胞骨架网络的分布构像及其中间纤维对Ｖｉｍｅｎｔｉｎ、Ｋｅｒａｔｉｎ两种抗体均呈现阳性反应。结论：恶性肿瘤细胞的中间纤维蛋白构型可能具有异质性的蛋白分子共表达，这对仅以ＴｒｉｔｏｎＸ－１００加（ＮＨ４）２ＳＯ４的抽提技术进行恶性肿瘤细胞中间纤维蛋白构型分析，是一个必需考虑的问题。     

钙调素拮抗剂TFP对Hela细胞微丝及波形纤维蛋白分布的影响

利用钙调素拮抗剂三氟拉嗪（ＴＦＰ）及微管的不同聚合状态对Ｈｅｌａ细胞微丝及波形纤维蛋白分布的影响进行了研究。Ｈｅｌａ细胞微丝明显解聚，而波形纤维蛋白则密集分布在细胞核一侧。细胞经ＴＦＰ处理２ｄ后，可见微丝分布的恢复，而波形纤维蛋白分布则变得分散，并向质膜延伸。但对ＴＦＰ处理２ｄ后的细胞用低温（２～４℃）处理，使微管解聚，或以紫杉酚（ｔａｘｏｌ）处理２４ｈ，使微管高度聚集以破坏其微管网络系统时，微丝则随之发生明显的解聚现象，而波形纤维蛋白又恢复为密集分布于细胞核一侧的状态。若在低温处理前加ｔａｘｏｌ预处理１．５ｈ，以稳定微管时，此时在细胞周边仍可见微丝存在，波形纤维蛋白仍保持其分散分布状态。结果表明经ＴＦＰ处理的Ｈｅｌａ细胞微丝及波形纤维蛋白分布的变化可能与微管分布的改变具有一定的联系。     

用酵母双杂合系统筛选与人血小板生成素受体c—Mpl相互作用 …

血小板生成素（ｔｈｒｏｍｂｏｐｏｉｅｔｉｎ,ＴＰＯ）是调节血小板生成最主要的细胞因子,其生物学效应由其受体ｃ－Ｍｐｌ介导,利用酵母双杂合系统（ｔｗｏ－ｈｙｂｒｉｄ ｓｙｓｔｅｍ）筛选与ｃ－Ｍｐｌ相互作用的蛋白质因子,以Ｇａｌ ４ＢＤ融合ｃ－Ｍｐｌ膜内部分ｃＤＮＡ的ｐＡＳＭＭ为靶蛋白质粒,筛选了人胎盘ｃＤＮＡ文库,分离到人波形纤维蛋白部分编码序列,首次检测到波形纤维蛋白与ＴＰＯ受体之间的相互作用,这     

不同S180细胞株蛋白质特性的电泳及免疫组化法比较研究

用ＳＤＳ聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）和免疫组化染色方法，比较研究４个不同单位保种的小鼠肉瘤（Ｓ１８０）细胞蛋白质的表达。电泳结果显示，北京市肿瘤研究所仲种的Ｓ１８０蛋白质条带数最多，武汉大学保种中心的Ｓ１８０最小。免疫组化染色法测定四个单位Ｓ１８０细胞的Ｒａｆ－１、Ｔｒｋ、Ｇａｉ、Ｇａｏ和ＮＦＫｂＰ６５蛋白表达，Ｇａｉ、ＴｒｋＡ和Ｒａｆ－１阳性率均低于０．２％；Ｇａｏ阳性率均低于４．２％     

两类微生物SOD的比较研究

报道了酵母ＳＯＤ工程菌ＺＨ－１／ｐＳＨ１比耐高温细菌Ｂ．Ｓ２１１－１５的抗Ｈ２Ｏ２水平高，其单位体积培养液和酵得率约为细菌的二倍，但单位细胞酶的含量比细菌低，高温细菌的ＳＯＤ热稳定性和存放稳定比酵母工程菌好，二者在ｐＨ４－１０范围内都很稳定，但对蛋白的耐受性差异很大，其中酵母ＳＯＤ工程勤务产好。     

Site-specific phosphorylation induces disassembly of vimentin filaments in vitro

Intermediate filaments are a major component of the cytoskeleton of eukaryotic cells. Although there appear to be at least five distinct classes of these filaments, cells of mesenchymal origin and most cells in culture contain the intermediate filament composed of the subunit protein vimentin. Vimentin exists in a nonphosphorylated as well as in a phosphorylated form, and there is increased phosphorylation of this protein when the filament undergoes marked redistribution in various cells. The role of phosphorylation on assembly-disassembly and organization of the vimentin filament has remained obscure. We report here a stable and purified system allowing biochemical examination of vimentin filament assembly and disassembly. Using this in vitro system, we carried out stoichiometrical phosphorylations, using purified protein kinases. We obtained evidence for site-specific, phosphorylation-dependent disassembly of the vimentin filament.     

Co-expression of vimentin and cytokeratins in parietal endoderm cells of early mouse embryo

Of the five classes of intermediate filaments found in vertebrate tissues, the cytokeratins are considered unique to epithelial tissues, while vimentin is expressed by endothelial and mesenchymal cells. In neither case is the precise function of the filament system known. Epithelial cells in culture often express vimentin as well as cytokeratins, but co-expression in vivo, as reported for pleomorphic adenomas of the parotid gland and metastatic carcinoma cells in ascites or pleural fluid, is still controversial. Here we report the co-expression of cytokeratins and vimentin in situ, in the parietal endoderm of the mouse embryo 8.5-13.5 days old. This population of individual, motile cells seems to be derived from a conventional epithelium by migration and differentiation. Our results support the idea that vimentin expression is specifically related to reduced cell-to-cell contact, and to the independent existence of a cell following detachment from an epithelial sheet.     

Redistribution of intermediate filaments during capping of lymphocyte surface molecules

Intermediate filaments (IF) constitute a major cytoplasmic filamentous network of higher eukaryotic cells that is distinct from actin and myosin microfilaments or microtubules. Although structurally similar, these filaments are formed by chemically and antigenically different proteins. Vimentin is the major IF polypeptide of mesenchymal cells and cultured non-mesenchymal cell lines. Recently, we have characterized a monoclonal IgM antibody from a patient with Waldenstr?m's macroglobulinaemia which is directed against vimentin. Using this monoclonal antibody, we have shown by direct immunofluorescence that intermediate filaments of human B and T lymphocytes consist of vimentin. In cells exposed to colcemid, the intermediate filaments retracted into a juxtanuclear aggregate ('coli') characteristic of vimentin filaments. As most components of the cytoskeleton, especially actin and myosin, have been implicated in the capping phenomenon, we investigated the effect of capping of either beta 2-microglobulin or membrane immunoglobulins on the organization of the intermediate filament network. We report that capping of these surface molecules induced the redistribution of vimentin just beneath the cap. When colcemid-treated cells were allowed to cap, the location of the cap always coincided with the coil, suggesting that the anchorage point of intermediate filaments is situated within the uropod.     

Coalignment of vimentin intermediate filaments with microtubules depends on kinesin.

Intermediate filaments in most types of cultured cells coalign with microtubules. Depolymerization of microtubules results in collapse of vimentin and desmin intermediate filaments to the nucleus where they form a perinuclear cap. Collapse can also be induced by microinjection of antibodies against intermediate filament or microtubule proteins. Thus, two filament systems interact with each other. But the molecules mediating this interaction are unknown. One of the candidates for this role is a microtubule motor kinesin. Recent data showed that kinesin is involved in the plus end-directed movement of the membranous organelles along microtubules such as radial extension of lysosomes in macrophages and centrifugal movement of pigment in melanophores. Here we report that injection of the anti-kinesin antibody into human fibroblasts results in the redistribution of intermediate filaments to a tight perinuclear aggregate but had no effect on the distribution of microtubules. Thus, kinesin is involved not only in organelle movement but also in interaction of the two major cytoskeletal systems, intermediate filaments and microtubules.     

鸭胚B淋巴细胞中间纤维的研究

从26天北京鸭胚胎的腔上囊中分离B淋巴细胞。应用细胞选择性系列抽提方法与DGD包埋——去包埋剂的电镜制样技术相结合,显示出了B细胞的中间纤维的超微结构及分布特点:纤维致密交织呈网络状,其特点是均匀地遍布于整个胞质空间,纤维单丝直径在9—11nm。间接免疫荧光实验结果表明,B细胞的胞质对波形蛋白单抗呈阳性反应,且也显示出了波形纤维呈网络状的结构形式。本文采用单向电泳、双向电泳和蛋白质免疫印迹技术,证实了B细胞中间纤维波形蛋白的分子量约为55KD。     

天然家蚕抗菌肽对人白细胞及巨噬细胞淋巴瘤细胞骨架系统作用的比较

报道了天然家蚕抗菌肽ＣＭ４对离体Ｕ９３７癌细胞骨架及核骨架损伤作用的扫描电镜观察。随着时间的延长，经天然家蚕抗菌肽ＣＭ４作用后的癌细胞骨架断裂，固缩成团状；癌细胞核骨架断裂，部分凝聚成团，结构不完整。相同剂量的天然家蚕抗菌肽ＣＭ４与正常人白细胞作用后细胞骨架及核骨架未见损伤现象。说明天然抗菌肽与重组抗菌肽的抗癌作用相同。     

A keratin cytoskeletal protein regulates protein synthesis and epithelial cell growth

Cell growth, an increase in mass and size, is a highly regulated cellular event. The Akt/mTOR (mammalian target of rapamycin) signalling pathway has a central role in the control of protein synthesis and thus the growth of cells, tissues and organisms. A striking example of a physiological context requiring rapid cell growth is tissue repair in response to injury. Here we show that keratin 17, an intermediate filament protein rapidly induced in wounded stratified epithelia, regulates cell growth through binding to the adaptor protein 14-3-3sigma. Mouse skin keratinocytes lacking keratin 17 (ref. 4) show depressed protein translation and are of smaller size, correlating with decreased Akt/mTOR signalling activity. Other signalling kinases have normal activity, pointing to the specificity of this defect. Two amino acid residues located in the amino-terminal head domain of keratin 17 are required for the serum-dependent relocalization of 14-3-3sigma from the nucleus to the cytoplasm, and for the concomitant stimulation of mTOR activity and cell growth. These findings reveal a new and unexpected role for the intermediate filament cytoskeleton in influencing cell growth and size by regulating protein synthesis.     

Control of germ cell nuclear behaviour at fertilization by Tetrahymena intermediate filament protein

Intermediate filament protein [relative molecular mass (Mr) 49,000 (49K)] from the ciliated protozoan Tetrahymena has been shown to resemble intermediate filament proteins from mammalian cells in several respects, and to have a possible role in the oral morphogenesis preceding binary fission in Tetrahymena. Here, based on immunofluorescence localization of the 49K protein in Tetrahymena during the early stages of conjugation, we suggest that the protein is involved in some nuclear events, such as the production of four haploid nuclei by prezygotic divisions (meiosis), selection of one of the four meiotic products, formation of the gametic pronucleus by the mitotic division of the selected meiotic product, transfer of the gametic pronucleus across a cell-cell junction, and zygote formation by pronuclear fusion.     

Abrogation of metastatic properties of tumour cells by de novo expression of H-2K antigens following H-2 gene transfection

H-2 gene transfection was used to restore expression of H-2K antigens in metastatic and non-metastatic subclones of a murine fibrosarcoma that lack their major histocompatibility complex-encoded H-2K antigens. De novo expression of H-2K reduced tumorigenicity and abolished the formation of metastasis in syngeneic mice. Expression of H-2K may lead to effective recognition of the disseminating tumour cells by the host immune system.