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1.
Collagen fiber formation in vitro was morphologically investigated on 22 epithelial-like cell lines originating from normal rat livers mainly by using specific histochemical stainings. All the cell lines, eight of which were cloned formed typical “collagenous” and/or “reticulin” fibers in the histological sense when they had been left as a full sheet for more than 3 weeks without subculture. Only one of five fibroblastic cell lines serving as control formed fibers to a lesser degree. Different patterns were recognized in fibers appearing between the liver cell cultures and the fibroblast cultures.  相似文献   

2.
Acid phosphatase, alkaline phosphatase, and lactic dehydrogenase activities have been compared in normal human diploid cell strains and in SV40-transformed heteroploid cell lines derived from them. A higher level of acid phosphatase activity was observed in diploid cultures derived from adult lung than in cultures derived from fetal lung of similar passage levels. The alkaline phosphatase activity of normal diploid fibroblasts was significantly higher than that of SV40-transformed cell lines derived from them. Generally, the lactic dehydrogenase activities of all these cell cultures were similar. Human diploid cells in culture “age,” in the sense that their ability to proliferate decreases with time during serial subcultivation. Evaluation of the activities of these three enzymes during the “aging” process showed that, although alkaline phosphatase and lactic dehydrogenase activities were similar in “young” and “senescent” cells, acid phosphatase showed a small but significant increase in the senescent cells.  相似文献   

3.
We characterized vegetative development in two inbred cell lines of Acetabularia acetabulum (L.) Silva. Cell growth occurred at the apex and by elongation of older interwhorls throughout vegetative development. Although cell length and hairs per whorl increased regularly during development, interwhorl length, hair persistence on the stalk, and complexity of each whorl (degree of branching of whorl hairs) showed sharp discontinuities during development in both cell lines. The first (earliest) discontinuity, formation of a short interwhorl, was the sixth interwhorl made in all cells. Even though cell line Aa1055 was twice the height ofAa4010 when mature, cells in both lines were 0.8–1.0 cm tall after formation of the short interwhorl. The second discontinuity, increases in hair persistence on the stalk and complexity of each whorl of hairs, began shortly before cap initiation. We propose the following nomenclature: 1) that slower growth before formation of the short interwhorl be called “juvenile”; 2) that more rapid growth after formation of the short interwhorl be called “adult”; and 3) that adult growth be separated into “early” and “late” phases by the discontinuities in whorl hair characteristics. The proposed developmental phases (juvenile, early adult, and late adult) are temporally sequential and spatially stacked.  相似文献   

4.
Chick embryo cell cultures release a particle-associated RNA-dependent DNA polymerase into the culture medium. The release shows a characteristic time course with a maximum on the 3rd or 4th day in culture. The release of enzyme decreases when the cells are further cultivated and passaged. The enzyme was characterized as an RNA-dependent DNA polymerase by its ability to transcribe heteropolymeric RNA into DNA. It is different from the polymerase of the avian leukosis/sarcoma virus group and indistinguishable from an RNA-dependent DNA polymerase from normal embryonated chicken eggs described previously [1, 2]. The release of enzyme is independent of the genetic systems regulating the complete or partial expression of the endogenous avian leukosis virus genome. The amount of enzyme released is dependent on the age of the embryo from which the cell cultures are prepared. Cells prepared from 6-day-old embryos release maximal enzyme activity.  相似文献   

5.
Chicken anemia agent (CAA) propagated in an established cell line derived from Marek's disease (MD) lymphoma (MDCC-MSB1). When passaged 19 times in MDCC-MSB1 cell cultures, it produced anemia of the same severity in chicks as it did before passage. Titration of the infectivity of CAA was performed successfully with subcultures of MDCC-MSB1 cell cultures which had been inoculated with serial tenfold dilutions of infected material. In it, no infected cultures could be subcultured. The propagation of CAA was also proved in the MD cell line, MDCC-JP2, and the avian lymphoid leukosis (LL) cell line, LSCC-1104B1, but not in the two MD cell lines, MDCC-RP1 and MDCC-BP1, or in the two LL cell lines, LSCC-1104X5 and LSCC-TLT. No CAA propagated in cell cultures prepared from skin and muscle, liver, or brain of chick embryos, or kidney, thymus, bursa of Fabricius, bone marrow, or white blood cells of chickens.  相似文献   

6.
The present study describes ontogenetic shifts in habitat use for 15 species of coral reef fish at Rangiroa Atoll, French Polynesia. The distribution of fish in different habitats at three ontogenetic stages (new settler, juvenile, and adult) was investigated in coral-dominated and algal-dominated sites at two reefs (fringing reef and inner reef of motu). Three main ontogenetic patterns in habitat use were identified: (1) species that did not change habitats between new settler and juvenile life stages (60% of species) or between juvenile and adult stages (55% of species—no ontogenetic shift); (2) species that changed habitats at different ontogenetic stages (for the transition “new settler to juvenile stage”: 15% of species; for the transition “juvenile to adult stage”: 20% of species); and (3) species that increased the number of habitats they used over ontogeny (for the transition “new settler to juvenile stage”: 25% of species; for the transition “juvenile to adult stage”: 25% of species). Moreover, the majority of studied species (53%) showed a spatial variability in their ontogenetic pattern of habitat use according to alternate reef states (coral reef vs algal reef), suggesting that reef state can influence the dynamics of habitat associations in coral reef fish.  相似文献   

7.
Early passage mouse embryo fibroblasts, mouse 3T3 cell lines, and early passage diploid human fibroblasts grew to higher cell densities in tissue culture medium supplemented with serum than in medium supplemented with defibrinogenated platelet-poor plasma (PPP). Unlike the mouse cells, the human fibroblasts displayed this differential growth response only in the presence of hypophysiologic concentrations of calcium. The addition of heat-treated extracts of human platelets to PPP-supplemented medium stimulated the replication of both the normal mouse cells and early passage human embryo fibroblasts. Human or mouse fibroblasts transformed by either retroviruses or by SV40, including SV40 infected “serum revertants” and “flat transformants,” grew to equal cell densities in medium supplemented with either serum or PPP. Infection of Balb/c-3T3 cells with SV40 rapidly induced them to grow in PPP-supplemented medium demonstrating that the ability of SV40-transformed cell lines to proliferate in PPP-supplemented medium does not arise from the cell culture selection procedures usually employed to obtain stable virus-transformed cell lines. 3T3 cells infected but not transformed by retroviruses do not replicate in PPP-supplemented medium demonstrating that reduction of the growth requirement for the platelet growth factor(s) by retroviruses is a transformation-specific response. Cell cultures that did not proliferate well in PPP-supplemented medium did not form tumors when inoculated into athymic nude mice. Many, although not all, of the lines which grew well in PPP medium were tumorigenic in nude mice. Together, these findings indicate that: (1) normal fibroblast-like cells display a growth requirement for factor(s) present in serum but not found in PPP; (2) this serum specific growth factor is derived from platelets; (3) a primary response to viral transforming genes is a reduction in the growth requirement for these platelet-derived factors; and (4) cells that have a reduced requirement for the platelet-derived growth factor are often tumorigenic.  相似文献   

8.
Spleen cultures prepared from mice injected 24 hr earlier with 2 × 106?2 × 108 sRBC and challenged in vitro with sRBC produced 10 times more anti-sRBC IgM PFC than cultures prepared from uninjected mice. The effect was specific for the particular species of foreign RBC injected in vivo. In vitro responses to TNP were also increased in spleen cultures prepared from animals injected 24 or 12 hr earlier with carrier RBC alone, directly implicating carrier-specific T cells in this process. Similar enhancements of PFC formation occurred in cultures prepared from mice which had been injected with sRBC 24 and 48 hr earlier, but which were exposed to lethal irradiation at 1 hr after injection of antigen, if their spleens were shielded extracorporeally during irradiation. This finding indicated that in vivo recruitment of antigen-reactive extrasplenic X-ray-sensitive cells from the circulating lymphocyte pool by the spleen could not account for the observed enhancement.Proliferation in the spleen of antigen-reactive T cells, commencing 12–20 hr after the administration of antigen, was demonstrated by the tritiated thymidine pulse technique. An 8-hr hot-pulse given to spleen cell cultures from normal animals at 20 hr after in vitro challenge with antigen did not affect the rate of generation of IgM-producing cells; however, administration of a similar pulse to cultures which were initiated at 12 or at 20 hr after the in vivo injection of sRBC eliminated the enhanced generation of PFC and delayed the in vitro response to sRBC by 24 hr.Spleen cell cultures were prepared from mice which had been injected in vivo with sRBC at 12, 20, and 70 hr earlier, and 8- to 10-hr hot pulses were given immediately after initiation of the cultures. The cultures were then challenged with sRBC-TNP; antibody responses to TNP were greatly reduced in hot-pulsed cultures prepared from mice injected in vivo with carrier RBC at 12 or 20 hr prior to initiation of the cultures. In contrast, antibody responses to TNP observed in hot-pulsed cultures prepared from mice which had been injected with carrier RBC at 70 hr prior to initiation of the cultures were generally similar to those of nonpulsed 70 hr control cultures. This result suggests that the onset of T helper cell proliferation begins within 12–20 hr after injection of antigen, but subsides in vivo within 70 hr. By that time, the antigen-reactive T cells have already differentiated to perform their helper function.In spite of the triggering of T-cell proliferation during the first 24 hr after injection of antigen, spleen cell cultures prepared from mice which had been injected 24 hr earlier in vivo with 2 × 108 sRBC produced only minimal numbers of anti-sRBC PFC if no antigen was added to the cultures. The presence of unprocessed antigen thus appears to be a requirement for B-cell proliferation in vitro, even after T-cell division has been triggered. This finding is consistent with earlier suggestions that the function of “helper” T cells may not be limited to passive transport of antigenic determinants to B cells. Evidence is also presented to support the contention that the antigen-reactive T cell involved in this process may have to undergo cell division in order to develop “helper” capacity.  相似文献   

9.
A machine was constructed, called a Cell Puller, that allows the steady advance or withdrawal of a microelectrode at very slow speeds—up to 170 μm/hr. Specially prepared microelectrodes held in the Cell Puller were placed in cultures of dissociated chick sensory ganglion neurons in such a way that growth cones attached to their tips. Movements of the microelectrodes, at speeds up to about 100 μm/hr, then resulted in the elongation of the neurites for up to 24 hr and for increases in length up to 960 μm; more rapid towing failed to cause extensive neurite elongation. Estimates of neurite diameter before and after “towing” indicated that a net increase in neurite volume had occurred. Furthermore, long neurites could be produced by towing from previously rounded neuronal cell bodies confined to small adhesive “islands” on a nonadhesive substratum. Neurites produced by microelectrode towing had a normal appearance, showed rapid saltatory movements of internal organelles and were capable of resuming growth on the substratum. Electron microscopy of bundles of neurites produced in this way from explanted dorsal root ganglia showed an ultrastructure typical of cultured neurites, with abundant longitudinally aligned microtubules and neurofilaments. These experiments demonstrate that neurites can grow in response to mechanical tension under tissue culture conditions. It is proposed that they do so also in normal development, the tension in this case being supplied initially by the locomotory activity of the growth cones and subsequently by the morphogenetic movements of the surrounding tissues.  相似文献   

10.
The term “contact inhibition of cell division” was borrowed from “contact inhibition of cell movement.” We prefer the term “postconfluence inhibition of cell division” as being more operational and less mechanistically biased; it is operationally defined as a pronounced depression of the mitotic rate in a postconfluent culture which displays a stationary density despite periodic nutrient renewal, the inhibition being locally reversibly by removal of the adjacent cells. The mechanism of postconfluence inhibition is of considerable interest because of the inverse correlation between postconfluence inhibition and the tumorigenicity of a number of cell lines. Several hypotheses, involving direct cell-to-cell contacts or locally restricted diffusion gradiens, could explain postconfluence inhibition. With the goal of discriminating among these hypotheses, time-lapse films were taken of carefully regulated, perfused cultures of 3T3 mouse cells, in which the transition from rapid growth to the stationary phase was recorded. Measurements of cell-to-cell contact, local cell density, and generation times were made on an individual cell level and analyzed with the aid of a computer. We observed that all-around cell-cell contact or a high local cell density present throughout G1 often did not produce immediate inhibition of cell division. We conclude that either (i) simple visible cell-cell contacts or a high local cell density are not the direct cause of postconfluence inhibition of cell division, or (ii) their effects often do not inhibit cell division until after a delay of about one cell generation time. Such a delay may be partly responsible for the 50% overshoot past the stationary density that we observed in 3T3 cultures.  相似文献   

11.
The aim of this work was to study the possible relationship between alkaloid production and growth measured as: biomass increase and cellular division frequency, in Datura stramonium in vitro root cultures (hairy root and normal cultures). A comparison of growth values on a fresh and dry weight basis showed that there were differences between transformed and non-transformed lines. The differential growth between lines occurred due to a real biomass increase and not because of water accumulation. On the other hand, the rate of cell division showed a similar pattern for all lines studied. Therefore, the differences in growth are not due to different cell division rates, nor to the presence of larger meristems, but to the development and growth of lateral roots and the presence of active intercalary meristematic zones in each line. The maximum alkaloid production occurred when the cultures were not growing. This suggests an inverse relationship. Finally, the data support a specific model of growth at the level of cell division in root cultures which has not been described before in the literature. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

12.
The expression of receptors for the Fc portion of IgG immunoglobin molecules was studied on tumor cell lines with high and low metastatic capacity. Two tumor cell lines from DBA/2 mice that had high metastatic activity, ESb and MDAY-D2, contained a high percentage of Fc receptor positive cells, as detected in a rosette assay with IgG antibody-coated erythrocytes (EA). In contrast, the low metastatic parental line Eb, from which ESb was derived, contained only a low percentage of EA-rosette-forming cells. ESb ascites tumor cells adapted to tissue culture in the presence of 2-mercaptoethanol (2ME) had a high expression of Fc receptors, whereas a cell line adapted to tissue culture in the absence of 2ME had a low expression of Fc receptors. “Soluble” Fc receptors were detectable by their ability to bind to EA and to cause blocking of rosette formation. They were found to be present in fluids from tumor-bearing animals, such as serum and cell-free ascites. Even animals with an ascites tumor of the low-metastatic line Eb contained “soluble” Fc receptors. The results are discussed with regard to their possible significance for tumor metastasis.  相似文献   

13.
The production of immunoglobulin by six cell lines derived from bursal tumors induced by avian leukosis virus follows two general patterns: (i) three cell lines that have been extensively passaged in culture synthesize and secrete light chains only; (ii) three cell lines that are recently isolated produce and secrete monomeric immunoglobulin M in addition to free light chains. All six cell lines synthesize and secrete both glycosylated and unglycosylated forms of light chain. We conclude that the cell lines established from lymphomas induced by avian leukosis virus represent relatively mature, but possibly abnormal, stages in the development of chicken B-lymphocytes. The immunoglobulin M produced by the cell lines failed to form detectable immune complexes with avian leukosis virus. It therefore appears that the immunoglobulin M is not directed against viral antigens and that autogenous antigenic stimulus cannot account for the sustained growth of the neoplastic B-lymphocytes.  相似文献   

14.
A total of 108 individuals from six different cultures found in the Department of Ica, Peru, were studied for the presence of Harris's Lines. Such lines or “bone scars” are formed due to cessation of bone growth due to episodes of starvation or illness and possibly other causes. These individuals covered a span of time of nearly 2,600 years. The individuals from mountain cultures had fewer lines and possibly a healthier childhood than those from coastal cultures. The modern population surveyed in this series still show a pattern of Harris's Lines similar to people from the Inca culture of 450 years ago.  相似文献   

15.
It has long been assumed that serial homologues are ancestrally similar—polysomerism resulting from a “duplication” or “repetition” of forms—and then often diverge—anisomerism, for example, as they become adapted to perform different tasks as is the case with the forelimb and hind limbs of humans. However, such an assumption, with crucial implications for comparative, evolutionary, and developmental biology, and for evolutionary developmental biology, has in general not really been tested by a broad analysis of the available empirical data. Perhaps not surprisingly, more recent anatomical comparisons, as well as molecular knowledge of how, for example, serial appendicular structures are patterned along with different anteroposterior regions of the body axis of bilateral animals, and how “homologous” patterning domains do not necessarily mark “homologous” morphological domains, are putting in question this paradigm. In fact, apart from showing that many so-called “serial homologues” might not be similar at all, recent works have shown that in at least some cases some “serial” structures are indeed more similar to each other in derived taxa than in phylogenetically more ancestral ones, as pointed out by authors such as Owen. In this article, we are taking a step back to question whether such assumptions are actually correct at all, in the first place. In particular, we review other cases of so-called “serial homologues” such as insect wings, arthropod walking appendages, Dipteran thoracic bristles, and the vertebrae, ribs, teeth, myomeres, feathers, and hairs of chordate animals. We show that: (a) there are almost never cases of true ancestral similarity; (b) in evolution, such structures—for example, vertebra—and/or their subparts—for example, “transverse processes”—many times display trends toward less similarity while in many others display trends toward more similarity, that is, one cannot say that there is a clear, overall trend to anisomerism.  相似文献   

16.
Goat serum: an alternative to fetal bovine serum in biomedical research   总被引:3,自引:0,他引:3  
Serum is frequently added to the defined basal medium as a source of certain nutritional and macromolecular growth factors essential for cell growth. Although a number of synthetic media have been prepared serum continues to be used in cell culture by many investigators. The best supplementation to a basal medium is fetal bovine serum (FBS) that is most frequently used for all types of cell cultures. During last four decades National Institute of Virology, Pune, has been working on isolation and identification of viruses from clinical specimens, employing tissue culture. Initially FBS was used for this purpose. However, due to its prohibitive cost and uncertain supply an alternative was sought. Commercially available sera from newborn calf, sheep, horse, human and serum obtained from goat blood (available from local abattoir) were tried. Goat serum (GS) was found to be suitable for most of the cell lines and primary cultures. Primary cultures from guinea pig embryo, monkey kidney, chick embryo, mouse peritoneal macrophages, and established cell lines were prepared and grown in growth media supplemented with GS. These cultures were studied for their morphology and growth in comparison with cultures grown in FBS containing media, and were used for mass cultivation of cells, quantitation and susceptibility of various virus strains, studies on effects of different nutrients and natural substances on cellular metabolism and virus replication, epitope analysis of various strains of Japanese encephalitis (JE) virus, strain differentiation studies, studies on antibody dependent plaque enhancement, assay of murine migration inhibition factor. Monoclonal antibodies against JE virus adapted to GS were characterised for their retention of functionalities. The results were comparable to those of cell cultures grown in FBS containing media. Similar results on chromosome studies were obtained from patient's whole blood cultures prepared in GS and FBS containing growth media. Organ cultures from mammalian, reptile and avian hosts; successfully grown in GS supplemented growth media, were used for different virological studies. Growth media supplemented with GS were used for in vitro cultivation of malarial parasites. Thus since the last three decades many scientists are using GS in place of FBS, in various fields of biomedical research. The present article reviews an account of the same.  相似文献   

17.
Listeriosis in sheep. Listeria monocytogenes excretion and immunological state in healthy sheep. Acta vet. scand. 1979, 20, 168–179. — The excretion of Listeria monocytogenes (Lm) in the faeces and milk, and humoral and cell mediated immunity against Lm, were examined in a sheep flock where no cases of listeriosis had occurred during the last 3 years. The investigation was carried out during the indoor season. During the first part of the season 2 of the 10 pregnant, 8 months old lambs excreted Lm in the faeces, but none of the 106 ewes, 2–10 years old. At lambing the organism was isolated from the faeces of 6 of the 10 1 year old lambs and from 64% of the ewes, and from the milk of 1 of the lambs and 41% of the ewes. Nearly all the isolates (98.5%) belonged to serotype 1. Antibody titres against Lm were found in sera and whey by an indirect haemagglutination method. The titres were higher for the ewes than for the hoggs and seemed to be influenced by the number of foetuses the animals carried. Cell mediated immunity was determined by a skin test where delayed hypersensitivity against an antigen prepared from Lm, was measured. Animals fed grass silage had a stronger reaction than animals fed hay, and a stronger reaction was found in animals with ≥ 3 foetuses than in the remainder. The investigation indicates that even in a healthy sheep flock all the animals may be exposed to Lm, and the majority may be latent carriers and excrete this organism in the faeces and milk during periods of stress.  相似文献   

18.
Evidence is presented that, contrary to common scientific “belief”, larva digeneans have profound effects on various components at various levels of the littoral marine ecosystem. Their ecological capacity includes: —Reduction of the breeding potential of host populations by “parasitic castration”; —Structural modification of host populations by generation of erratic growth patterns, size-class differential mortality and “negative growth”; —Induction of host-population mortality and control by increased susceptibility to environmental stress; —Induction of changes in community structure by removal of hosts from their normal trophic levels; —Interference with major energy-flow pathways by precocious removal of hosts from their normal food-web position; —Interference with host-biomass, production and turnover-rate estimations by by-passing of hostassimilated energy; —Interference with predator-prey systems by affecting either component(s) of such systems.The notorious neglect of these factors by marine ecologists and, even more, their total unawareness of the effects these factors produce, raise serious doubts about the validity of marine ecological data and concepts. For the parasitologist, on the other hand, the study of ecological aspects of marine parasite (digenean) biology may open new avenues of research. With a true synthesis of both scientific disciplines, we may eventually arrive at a point where “more complete knowledge of life cycles will permit more intelligent and more effective regulatory methods, the reduction of morbidity, the advancement of health, and the conservation of natural resources.”The latter statement has not been cited from a recent issue of a scientific journal; it has been written down as long as 46 (!) years ago by one of the most outstanding investigators of marine digenean life cycles—Horace W. Stunkard (1940, p 15), but has lost nothing of its actuality. It is hoped that Stunkard's far-sighted words might encourage parasitologists to devote some of their scientific power and skill to the study of marine ecoparasitological problems—for the sake of a better understanding of ecological processes and to the benefit of the endangered marine life.  相似文献   

19.
唐明  汪超  谭韵雅  李群 《广西植物》2016,36(5):582-588
为提高灰毡毛忍冬"渝蕾1号"悬浮培养体系中绿原酸的含量,该研究探讨了B_5培养基中不同浓度的无机盐对灰毡毛忍冬"渝蕾1号"悬浮培养细胞生物量及绿原酸含量的影响,通过在悬浮培养体系中添加不同浓度的无机盐,采用重量法测定灰毡毛忍冬"渝蕾1号"悬浮培养细胞的生物量及采用高效液相色谱法测定绿原酸的含量。结果表明:当硝态氮和铵态氮配比与B_5培养基中硝态氮和铵态氮配比一致时,即NO_3~-/NH_4~+摩尔比值为13∶1时,培养体系有利于细胞的生长和绿原酸的积累。当KNO_3浓度为3.5 g·L~(-1)时,细胞生物量达到最大,为19.26 g·L~(-1);当KNO_3在较低浓度(0.5 g·L~(-1)和1.5 g·L~(-1))时,积累较多的绿原酸。NO_3~-的两项研究结果均与对照浓度(2.5g·L~(-1))有一定的差异。另外,对(NH_4)_2SO_4来说,在高于对照浓度0.134 g·L~(-1),即浓度为0.268 g·L~(-1)时,生物量和绿原酸含量都达到了最大。P、Ca、Mg三种矿质元素的研究结果表明,当Na H_2PO_4·2H_2O浓度为0.10 g·L~(-1)、Ca Cl_2的浓度为0.20 g·L~(-1)时,细胞的生长和绿原酸的积累均可达到最大值;而对Mg~(2+)来说,低浓度促进细胞的生长,高浓度促进绿原酸的积累。兼顾细胞生物量和绿原酸含量两个指标,需选择适中的浓度。这些结果均与对照浓度有一定的差异。这说明灰毡毛忍冬"渝蕾1号"悬浮细胞所需无机盐的浓度与B_5培养基无机盐的浓度有一定的差异,选择适宜的浓度可促进其悬浮细胞的生长及次生代谢产物绿原酸的积累。该研究结果为绿原酸的工业化生产打下了基础。  相似文献   

20.
Cultured primary cells exhibit a finite proliferative lifespan, termed the Hayflick limit. Cloning by nuclear transfer can reverse this cellular ageing process and can be accomplished with cultured cells nearing senescence. Here we describe nuclear transfer experiments in which donor cell lines at different ages and with different proliferative capacities were used to clone foetuses and animals from which new primary cell lines were generated. The rederived lines had the same proliferative capacity and rate of telomere shortening as the donor cell lines, suggesting that these are innate, genetically determined, properties that are conserved by nuclear transfer.  相似文献   

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