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1.
2.
The potential calcium-binding protein p9Ka is related to S-100 protein and the vitamin D-dependent intestinal calcium-binding protein. p9Ka accumulates abundantly in cultured rat mammary myoepithelial-like cells but is very much less abundant in the parental cuboidal epithelial cells. p9Ka mRNA is found in normal rat mammary gland, and preliminary experiments suggest that it is found in the mammary myoepithelial cells. A 17-kilobase pair fragment of cloned normal rat DNA contains the gene for p9Ka, but it also contains the gene for two additional polypeptides of molecular mass 6 kDa that are resolved as two isoelectric focusing variants by two-dimensional gel electrophoresis. These two isoelectric focusing variants correspond to two abundant polypeptides present in the cultured myoepithelial cells and probably arise from postsynthetic modification of the product of a single gene. The mRNA for the product of this gene and the p9Ka mRNA are both found in the normal rat mammary gland, but these two mRNAs are differentially expressed in certain tumor-derived rat cell lines. 相似文献
3.
Control of type IV collagen production in rat mammary epithelial and myoepithelial-like cells 总被引:1,自引:0,他引:1
M J Warburton R Kimbell P S Rudland S A Ferns R Barraclough 《Journal of cellular physiology》1986,128(1):76-84
A rat mammary myoepithelial-like cell line (Rama 401) produces 3.5 times more type IV collagen than a mammary epithelial cell line (Rama 25), as measured by the formation of protein hydroxyproline. However, using quantitative "dot" hybridization techniques, the level of poly (A)-containing mRNA hybridizing to a type IV collagen cDNA probe is only 50% higher in Rama 401 cells than in Rama 25 cells. The total amount of hydroxyproline synthesized per cell by the two cell lines is similar. However, in the Rama 25 cells approximately 70% of the hydroxyproline is found as free hydroxyproline against 13% for Rama 401 cells. When Rama 25 cells are grown on collagen gels, they accumulate 2.5-fold more type IV collagen. However, type IV collagen mRNA levels are only 30% higher in Rama 25 cells grown on collagen. The total amount of hydroxyproline synthesized is the same as cells grown on plastic, whereas the extent of collagen degradation is reduced from 71% to 30% in cells grown on collagen gels. No degradation of type IV collagen can be detected in the culture medium of Rama 25 cells. These results indicate that the increased accumulation of type IV collagen in Rama 401 cells is not due to increased synthesis but to a decreased rate of intracellular degradation, and that for Rama 25 cells, the extracellular matrix modulates type IV collagen production by regulating the rate of intracellular collagen degradation. 相似文献
4.
Different growth factors stimulate cell division of rat mammary epithelial, myoepithelial, and stromal cell lines in culture 总被引:2,自引:0,他引:2
A number of single-cell-cloned cell lines have been used to examine the growth-promoting effects of putative mammotrophic agents on the various cell types in normal and neoplastic rat mammary glands. A partially purified novel pituitary-derived growth factor stimulates only cuboidal epithelial cells to divide whereas fibroblast growth factor (FGF) stimulates the growth of stromal and myoepithelial-like cells. Epidermal growth factor (EGF) has a widespread but variable growth-stimulating action, but prolactin and growth hormone are essentially inactive when added alone at a concentration of 5 micrograms/ml. Phosphoethanolamine stimulates the growth of one epithelial cell line and a derivative myoepithelial-like cell line, but is inactive on the other cell lines tested. The use of defined cloned cell lines provides a direct and reproducible assay for the identification and purification of inducers of mammary growth. 相似文献
5.
Philip S. Rudland Christine M. Hughes Sharon A. Ferns Michael J. Warburton 《In vitro cellular & developmental biology. Plant》1989,25(1):23-36
Summary Parenchymal organoidal structures that were obtained from collagenase digestion of reduction mammoplasty specimens of apparently
normal human breasts have been grown in short-term primary cultures, either on plastic or on floating gels of polymerized
rat-tail collagen. Three morphologically distinct major cell types are readily observed in both systems: cuboidal cells, which
occupy apical positions on collagen gels; larger, epithelioid, or basal cells on gels; and elongated cells which penetrate
into the gel. In addition, a fourth cell type, that of a large, flat cell, is observed less readily by phase contrast microscopy
on the surface of cultures grown on plastic. Immunofluorescent and immunocytochemical staining of cultures on plastic or histologic
sections of cultures on gels have been undertaken with antisera and other histochemical reagents that stain the different
parenchymal cell types in vivo. Thus antisera to epithelial membrane antigen(s), monoclonal antibodies (MABs) to the defatted
mammary milk fat globule membrane, peanut lectin, and keratin MAB LE61, which preferentially stain the epithelial cells of
ducts in vivo, also stain the cuboidal/apical cells in vitro. The large, flat cells are stained intensely by the first three
reagents but not by the last one. Antisera to collagen IV, laminin, fibronectin, actin, keratin MAB LP34, MABs to the common
acute lymphoblastic leukemia antigen, and MAB LICR-LON-23.10, which showed enhanced staining for the ductal myoepithelial
cells in vivo, also stain the epithelioid/elongated cells in vitro. However, the effect of the last four reagents is reduced
considerably in most elongated cells, and MAB LP34 stains the large, flat cells intensely. Heterogeneous cells of intermediate
morphologies and staining patterns between the cuboidal/flat cells and large epithelioid cells have also been identified.
The results suggest that the cuboidal cells and large, flat cells are related to mammary epithelial cells, whereas the large
epithelioid/elongated cells have some characteristics of myoepithelial cells, and that intermediate forms may exist in culture
between the two parenchymal cell types.
This work was supported in part by the Ludwig Institute for Cancer Research and the Cancer and Polio Research Fund. Dr. M.
J. Warburton is supported by the Cancer Research Campaign. 相似文献
6.
Peroxidase-conjugated Griffonia simplicifolia-1 (GS-1) and pokeweed mitogen (PWM) histochemically stain only the myoepithelial cells and not the epithelial or fibroblastic cells of rat mammary glands preserved in methacarn or glutaraldehyde and embedded in paraffin. This pattern of staining occurs in other rat exocrine glands except the pancreas, but is the reverse of that seen in most lining epithelium. The histochemical binding of GS-1 and PWM to myoepithelial cells is inhibited specifically by D-galactose and by polymers of N-acetylglucosamine, respectively. GS-1 and its subcomponent, GS-1-B4, also bind to extracellular structures similar to those stained by anti-laminin serum. At the ultrastructural level, both conjugated GS-1 and PWM bind to the plasma membrane of the myoepithelial cells, as well as to the adjacent basement membrane. Non-metastasizing rat mammary tumors produced by dimethylbenz[a]anthracene, by derivative epithelial stem-cell lines, and by a transplantable tumor all contain more elongated myoepithelium-like cells as well as cuboidal epithelium-like cells; both cell types are neoplastic. The more elongated myoepithelium-like cells are stained by GS-1 and PWM, whereas the cuboidal epithelium-like cells are unstained. Moderately and strongly metastatic rat mammary tumors produced by epithelial cell lines and by transplantable tumors, respectively, contain no such neoplastic cells that bind either lectin. We suggest that the carbohydrate receptors for GS-1 and PWM are consistent markers for the presence of the myoepithelial cell in normal and tumorous rat mammary glands. 相似文献
7.
Joanne Mcandrew Philip S. Rudland Angela M. Platt-Higgins John A. Smith 《The Histochemical journal》1994,26(4):355-366
Summary Immunoreactive alpha-transforming growth factor (-TGF) was shown by immunocytochemistry to be present in the rat mammary gland at various stages of development, the staining being most intense in mature myoepithelial cells. -TGF was also detected in the secretions of the mammary glands of pregnant and lactating rats. -TGF in the extracts of rat mammary glands at each stage of development, and in several rat mammary cell lines and in culture medium in which they had been grown, was shown by Western blotting to consist primarily of a protein of molecular weight 50 kDa. The amount of this protein was greater in the mammary gland of the lactating rat than in resting or involuting glands. -TGF was also found in some, but not all, human breast carcinomas, and in benign hyperplastic breast diseases. 相似文献
8.
OBJECTIVE--To define usual colour and site of storage of visiting bags in general practitioners'' cars and to investigate effect of these variables on temperature inside bag. DESIGN--Questionnaire to general practitioners; serial temperature measurements from paired black visiting bags at different storage sites and from bags of different colour. SETTING--South Devon coastal town during May and June. SUBJECTS--200 general practitioners, of whom 145 returned legible questionnaires. MAIN OUTCOME MEASURES--Bag colour, duration and site of storage, temperature inside black bags at defined storage sites, and effects of bag colour on internal temperature. RESULTS--111 (77%) of the general practitioners carried a black visiting bag, and 76 kept their bag in their car all day. The bag was coolest in the car boot, but irrespective of storage site, maximum internal temperature of the bag was always over 25 degrees C and reached up to 80 degrees C. Spraying a black bag silver significantly reduced the bag''s internal temperature (mean difference 8.37 degrees C (95% confidence interval 6.68 to 9.86 degrees C) df = 59, t = 10.29, P < 0.001). CONCLUSIONS--General practitioners should use a silver coloured visiting bag; when visiting, they should store it in their car boot; at other times they should remove it to a cooler site. 相似文献
9.
Distribution of entactin in the basement membrane of the rat mammary gland. Evidence for a non-epithelial origin 总被引:4,自引:0,他引:4
M J Warburton P Monaghan S A Ferns P S Rudland N Perusinghe A E Chung 《Experimental cell research》1984,152(1):240-254
Entactin, a sulfated glycoprotein with a molecular weight (MW) of about 150 kD, is present in vascular basement membranes and in the interstitial connective tissue of the mammary glands of virgin rats. It does not appear to be present in the basement membrane surrounding the mammary ductal system. However, in lactating mammary glands entactin is also present in the basement membrane region surrounding the secretory alveoli. Ultrastructural localisation of entactin reveals that it is present on the basal surface of epithelial cells, with patchy staining in the lamina lucida and lamina densa. Entactin also appears to be associated with interstitial collagen fibres. Mammary fibroblastic cells in culture are able to produce entactin, whereas mammary epithelial and myoepithelial cells, which synthesise the basement membrane proteins laminin and type IV collagen, fail to synthesise entactin. 相似文献
10.
Minnie K. O''Farrell Dorothie Clingan Philip S. Rudland Luis Jimenez De Asua 《Experimental cell research》1979,118(2):311-321
The ability of prostaglandin F2α (PGF2α) and other prostaglandins to stimulate the initiation of DNA synthesis in quiescent cultures of various mouse fibroblastic cell types has been investigated. PGF2α was found to be more effective than the other prostaglandins. Most cell types, with the exception of BALB/c 3T3, responded to PGF2α. Addition of PGF2α in combination with insulin resulted in a synergistic increase in the proportion of cells synthesizing DNA. The effect of nutrients on the stimulation of the initiation of DNA synthesis has been examined in detail; it was found that Swiss 3T3 cells showed a requirement for hypoxanthine and vitamin B12 whereas Swiss 3T6 cells demonstrated a stringent requirement for vitamin B12 only. The effect of prostaglandin precursors, synthetic analogues of the prostaglandin endoperoxides and inhibitors of prostaglandin synthesis was also examined in two cell types. The effect of PGF2α was compared with that of two polypeptide growth factors, epidermal growth factor (EGF) and fibroblast growth factor (FGF) in Swiss 3T6 cells grown in 0.0025% (v/v) serum. In combination with insulin each of these three growth factors stimulated the initiation of DNA synthesis in approximately the same number of cells. 相似文献