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1.
We examined effects on intestinal absorption of cholesterol and triglycerides and intestinal lipoprotein formation by feeding rats diets in which saturated fatty acids (palmitic plus stearic) comprised 78%, 68%, 48%, or 38% of triglyceride fatty acids. Absorption into lymph of radiolabeled cholesterol was proportional to triglyceride absorption. The rates of absorption of these lipids were related inversely to the % saturated fatty acids fed. The distribution of newly absorbed cholesterol and triglyceride into intestinal lipoproteins differed. With increasing cholesterol absorption more was recovered in very low density lipoproteins in contrast to the appearance preferentially in chylomicrons of larger quantities of fatty acid. Lymph lipid content did not reflect a consistent pattern in relation to the experimental diet fed. The fatty acid composition of triglyceride-rich lymph lipoproteins resembled the diet closely. One-quarter of the intestinal lymph particles from rats fed the highly saturated diets was flattened and polygonal as judged by electron microscopy if cooled to room temperature; whereas with the same diets, particles collected and isolated at 37 degrees C were round. Proportions of A-I and C apolipoproteins in triglyceride-rich intestinal particles varied inversely; apoA-I increased as fat/cholesterol absorption was greater. Diet-induced alterations in plasma lipoproteins and increased circulating triglycerides in this study in rats were unrelated to the variations in intestinal absorption or lymph lipoprotein formation.  相似文献   

2.
Hepatic lipid droplets. Isolation, morphology and composition   总被引:4,自引:1,他引:3       下载免费PDF全文
The floating lipid layer isolated centrifugation of rat liver was examined for composition and ultrastructure. It was chiefly composed of triglycerides and cholesterol esters plus much smaller amounts of free cholesterol, diglycerides, phospholipid and protein. No free fatty acids were detected. The triglyceride and cholesterol ester fractions consisted mostly of esters of linoleic acid, oleic acid and palmitic acid. Electron micrographs of the floating lipid layer revealed numerous spherical osmiophilic droplets having a mean diameter of 0.5-2mum with a very-thin dense outer coat. Similar structures were observed as organelles in electron micrographs of the intact liver cell. The amount of triglyceride in the layer decreased in rats starved for 72h, but pellet triglyceride (homogenate minus the floating lipid layer) was unchanged. These results suggest that the floating lipid layer is the representative in vitro of lipid-rich organelles which probably function as a depot form of hepatic-cell neutral lipid.  相似文献   

3.
A reproducible, fairly narrow-sized population of rat lymph chylomicrons, approximately 100 nm, was isolated by centrifugation and combined with low levels of [1-13C]oleic acid for NMR studies. The carboxyl chemical shift was monitored as a function of aqueous pH to characterize the ionization behavior of the fatty acid in these particles. The titration curves were very similar to those for oleic acid in equivalent-sized emulsion particles composed of egg phosphatidylcholine and triolein. A simple partition-ionization model was fitted to the data to derive values for apparent ionization constant, expressed as pKapp, of 7.4-7.5 and the "true" surface to core partition coefficient of approximately 7 for oleic acid in chylomicrons. The fatty acid in chylomicrons thus appeared to be largely associated with the surface regions of these particles. Addition of bovine serum albumin to the samples showed that near physiologic pH much of the fatty acid was bound to the albumin at fatty acid to albumin-binding stoichiometries as high as 5.1 and with mass ratios of greater than 2 in favor of the lipid or lipoprotein particles. Lowering the pH of the medium shifted the distribution of fatty acid away from albumin so that at pH 5 with the emulsion, virtually all the fatty acid was associated with the lipid. The behavior observed under physiologic conditions is consistent with the rapid clearance and redistribution of fatty acid generated in these particles by lipolytic processes. However, under conditions of severe acidosis, hyperlipidemia, and hypoalbuminemia a significant portion of fatty acids might be retained in triglyceride-rich lipoproteins and their remnants and affect subsequent metabolism.  相似文献   

4.
The uptake of lipids by rat liver cells   总被引:6,自引:3,他引:3       下载免费PDF全文
1. Unesterified cholesterol, cholesterol esters and triglycerides of chylomicrons were taken up at the same rate by isolated hepatic parenchymal cells. 2. On incubation of hepatic cells, isolated 2min. after the injection of chylomicrons in vivo, the chylomicron triglyceride associated with the cells underwent hydrolysis. 3. In cells isolated 5min. after the injection of chylomicrons, the chylomicron triglyceride bound to the hepatic cells was accessible to added clearing factor lipase. 4. ;Ghost' hepatic cells had the same binding capacity and lipolytic activity per cell as intact cells. 5. Of all subcellular fractions studied, the ;plasma membrane' fraction showed the greatest capacity per unit weight for non-esterified fatty acid and chylomicron triglyceride binding and for triglyceride hydrolysis. 6. Once non-esterified fatty acids entered the hepatic cell, they were apparently metabolized in the same manner, whether taken up from the circulation as such or derived from chylomicron triglyceride.  相似文献   

5.
The surface coat of chylomicrons: electron microscopy   总被引:4,自引:0,他引:4  
Electron microscope studies were performed on thoracic duct lymph and on washed chylomicrons from dogs fed corn oil. High-resolution electron micrographs showed the presence of a surface coat that differed from the core material and did not resemble a plasma membrane. This was true for both chylomicrons in whole lymph and those that had been subjected to repeated washing. Apparently, the chylomicrons, while passing from the intracellular to the extracellular space, do not acquire their surface coat from pinched off cellular membrane.  相似文献   

6.
The study was undertaken to investigate the metabolic rat of post-lipolysis mesenteric lymph small chylomicrons produced in vitro. Small chylomicrons doubly labeled with [3H]cholesterol (more than 70% in cholesteryl esters) and [14C]palmitate-labeled triglycerides were collected from rat mesenteric lymph during periods of fasting. Lipolysis was performed in vitro with lipoprotein lipase purified from bovine milk. More than 98% of the chylomicron-triglycerides could be hydrolyzed to fatty acids. Post-lipolysis chylomicrons were separated by zonal ultracentrifugation, characterized, and tested for biological behavior in intact rats. Following lipolysis the lipoproteins lost nearly all their triglycerides, apoA-I, and apoC, and were relatively enriched with cholesteryl esters, unesterified cholesterol, phospholipids, and apoB. Three preparations were tested for biological behavior: pooled (total) post-lipolysis chylomicrons (diameter approximately 250 A); particles at the ascending part of the zonal effluent (diameter approximately 300 A), and at the descending part (diameter approximately 200 A). After intravenous injection to intact rats, [3H]cholesteryl ester decay was very rapid with pooled lipoproteins and the 300-A preparation (t1/2 = 5-10 min). The 200-A preparation in contrast stayed in circulation much longer (t1/2 = 60-90 min). The study thus demonstrated metabolic heterogeneity of post-lipolysis small chylomicrons and indicated that some may form an LDL-like subpopulation with a plasma lifetime slower than "remnants" but faster than LDL.  相似文献   

7.
A comparison was made between the effects of chylomicrons and chylomicron remnants on metabolic processes of isolated hepatocytes. Since isolated triacylglycerol-rich lipoproteins are contaminated with nonesterified fatty acids, control incubations were conducted with an amount of fatty acid equivalent to the contaminating fatty acids present in the chylomicrons and the remnant preparations, respectively. Chylomicron remnants, produced in vitro by incubation of chylomicrons in postheparin rat plasma, caused marked inhibition of glycolysis, fatty acid synthesis, and cholesterol synthesis, along with marked stimulation of ketogenesis. These effects were traced to the release of nonesterified fatty acids from these remnant particles as a consequence of contamination with lipoprotein lipase, picked up by the particles during the incubation with rat plasma. Fatty acids inhibit glycolysis, cholesterol, and fatty acid synthesis, but enhance ketone body formation by isolated hepatocytes. Chylomicrons and remnants prepared in vivo by the injection of chylomicrons into functionally hepatectomized rats were not contaminated with lipoprotein lipase and did not inhibit glycolysis and cholesterol synthesis nor increase ketone body formation. These lipoprotein particles did, however, cause significant inhibition of fatty acid synthesis, with the chylomicrons being more effective on a protein basis than the remnants produced in vivo. The mechanism responsible for the inhibition of fatty acid synthesis by chylomicrons and remnants prepared in vivo remains to be resolved.  相似文献   

8.
EFFECTS OF LIPOPROTEIN LIPASE ON THE STRUCTURE OF CHYLOMICRONS   总被引:3,自引:0,他引:3       下载免费PDF全文
Chylomicrons isolated from rat lymph were complexed with lipoprotein lipase of post-heparin plasma (chylomicrons-LPL) in order to study the effects of lipolysis on the structure of chylomicrons. Triglyceride in the chylomicron core was readily hydrolyzed to free fatty acids (FFA) and glycerol when chylomicrons-LPL were incubated at pH 8.3 in medium containing albumin. Although most of the FFA were immediately released to the medium, some were retained within chylomicrons when FFA-binding sites on albumin were not available. These observations suggest that albumin may have a specific role in the transfer of FFA across the chylomicron surface film. Chylomicrons-LPL assumed many different shapes as they were depleted of triglyceride by the lipolytic action of the enzyme, and total removal of core triglyceride resulted in empty sacks of surface film. The surface film was visualized in sections of OsO4-fixed chylomicrons-LPL as a thin electron-opaque line, 25–30 Å wide, in areas where the underlying electron-opaque core had been replaced by zones of decreased electron opacity, and in folds of surface film extending outward from chylomicrons partially depleted of core lipid. The findings demonstrate that chylomicrons consist of a core of liquid triglyceride enveloped by a pliable and durable monolayer surface film, and that lipoprotein lipase reduces the triglyceride core without disrupting the surface film.  相似文献   

9.
A technique for freezing ram and bull spermatozoa in pellet form, using the cold surface of cattle fat was compared to other freezing procedures. Three freezing methods were compared to cryopreserve ram spermatozoa: 0.25 ml straws, pellets frozen on the cold surface of paraffin wax and pellets frozen on the cold surface of cattle fat. In addition, two cryoprotectants, glycerol or sucrose, in an egg yolk-Tris diluent were compared. Ram spermatozoa frozen as pellets on cattle fat exhibited higher percentages of motile cells after thawing (54%) than spermatozoa frozen in straws (49%) or as pellets on paraffin wax (42%, S.E.M. = 1; P < 0.05). However, the percentages of acrosome intact cells were similar for spermatozoa frozen as pellets (49%) and spermatozoa frozen in straws (48%; P > 0.05), but higher than for spermatozoa frozen as pellets on paraffin wax (39%, S.E.M. = 1; P > 0.05). Ram spermatozoa exhibited higher percentages of motile cells after thawing when the cryoprotectant was sucrose (51%) compared to glycerol (46%; P < 0.05). Similarly, acrosomal integrity was greater with sucrose (49%) than with glycerol (42%; P < 0.05). Bull spermatozoa exhibited higher percentages of motile cells after thawing, when cells were frozen in straws (47%) than in the pellet form, regardless of the surface on which the pellets were frozen (31-37%, S.E.M. = 3; P < 0.05). However, bull spermatozoa exhibited higher percentages of motile cells when frozen as pellets on the surface of cattle fat (66%) or dry ice (61%), than when frozen on paraffin wax (53%, S.E.M. = 4; P < 0.05). In conclusion, although bull spermatozoa survive cryopreservation more effectively in straws, ram spermatozoa can be cryopreserved as pellets on the cold surface of cattle fat using sucrose as the cryoprotectant. This technique is simple, requires little equipment, is less expensive than using straws and may prove useful for cryopreserving ram and possibly bull spermatozoa in developing countries.  相似文献   

10.
Fatty acids are important intermediate molecules in lipid metabolism. During lipolysis of intracellular lipid droplets or plasma triacylglycerol-rich lipoproteins, fatty acids are generated and may transiently accumulate. We therefore studied the distribution of both fatty acid and free cholesterol between the core and surface of phosphatidylcholine-triolein emulsions at pH 7.4. Nine emulsion systems containing 0.8 to 6.6% cholesterol and 0.16 to 1.02% oleic acid were formed, and core and surface phases were isolated. Phospholipid distributes only to the surface phase. The distribution coefficient of cholesterol surface to core was 23.9 +/- 3.6 S.D., i.e., there was approx. 24-times more cholesterol per unit mass in the surface than in the core phase. This distribution was unchanged by the presence of different quantities of fatty acid in the emulsion particles. The apparent distribution coefficient of fatty acid in surface to core was about 10 at low cholesterol contents and fell to about 7 at high cholesterol contents. However, when the apparent distribution coefficient of fatty acid was related only to the phospholipid component of the surface, the apparent distribution coefficient was constant at about 12.3 +/- 1.1 S.D. Since the fatty acid in the surface phase is about half ionized the true distribution coefficient of unionized fatty acids is about 6.2. The results indicate that fatty acids partition into the phospholipid domains of the surface and not into cholesterol domains and the distribution of fatty acids into surface phospholipid domain is not affected by cholesterol content.  相似文献   

11.
The effects of filler used in the pellet cores (ie, waxy cornstarch or lactose) and the enteric film coat thickness on the diffusion and dissolution of a freely soluble drug were studied. Two kinds of pellet cores containing riboflavin sodium phosphate as a model drug, microcrystalline cellulose (MCC) as a basic filler, and waxy cornstarch or lactose as a cofiller were film coated (theoretically weight increase 20% or 30%) with an aqueous dispersion of cellulose acetate phthalate (CAP). The diffusion of riboflavin sodium phosphate in aqueous enteric-coated pellets was investigated using noninvasive confocal laser scanning microscopy (CLSM). The in vitro release tests were performed using a USP apparatus I (basket method). Diffusion of drug from the core to the film coat was found to be greater with lactose-containing pellets than with waxy cornstarch-containing pellets. The dissolution test showed that 30% enteric-coated waxy cornstarch pellets had a good acidic resistance in 0.1 N HCl solution for at least 1 hour, while the other enteric pellet formulations failed the test. The waxy cornstarch-containing enteric pellets dissolved at SIF in less than 10 minutes. Confocal images of film-coated pellets showed that waxy cornstarch-containing pellets had less drug dissolved than respective lactose-containing pellets. The observations were further confirmed by measurement of fluorescence intensity of riboflavin sodium phosphate in the film coat. The dissolution test was consistent with the confocal microscopy results. In conclusion, waxy cornstarch as a cofiller in the pellet cores minimizes premature drug diffusion from the core into the film coat layer.  相似文献   

12.
1. The hepatic metabolism of chylomicrons and chylomicron remnants was compared after adding approximately equal numbers of each lipoprotein particle to the perfusate of isolated livers. 2. At least 40% of the added remnants were metabolized by the liver compared with less than 3% for chylomicrons. 3. There was significantly more net removal of labelled remnants than of chylomicrons by the liver. 4. A greater proportion of labelled cholesterol than of labelled triacylglycerol fatty acids was transferred to the liver from each lipoprotein. 5. Cholesteryl esters of remnants were hydrolysed to triacylglycerol fatty lipoprotein. 5. Cholesteryl esters of remnants were hydrolysed to triacylglycerol fatty acids of remnants were oxidized to CO2 more extensively than those of chylomicrons. 6. There was greater oxidation of remnant glycerolipic [(1(-14)C]oleate than of glycerolipid [1(-14)C]palmitate. 7. A large fraction of the fatty acids of remnants, but not of chylomicrons, was transferred to phospholipids, which were released by the liver in a lipoprotein of relative density less than 1.006. 8. Label from remnants, but not from chylomicrons, was found in lipoproteins of relative density greater than 1.006, which were not released during perfusion but could be flushed out from the liver at the end of perfusion.  相似文献   

13.
The presence of noncovalently associated lipids and covalently bound fatty acids was investigated in preparations of mucus glycoproteins obtained by using density-gradient centrifugation in CsCl/guanidinium chloride. No phospholipids, glycolipids, cholesterol, or triglycerides could be detected. However, small amounts of extractable fatty acids were consistently found, the sum of which ranged from 0.3 to 0.9 micrograms/mg of glycoprotein. The amount of fatty acid released after subsequent treatment with KOH ranged from 0 to 27 ng/mg of glycoprotein. We conclude that density-gradient centrifugation in CsCl/guanidinium chloride is very efficient in removing noncovalently associated lipids from mucus glycoproteins and that covalently bound fatty acids are probably not present in the macromolecules.  相似文献   

14.
Lymph fistula rats were continuously infused with emulsions containing diacylglycerol consisting of 1,3-species (65.6%), 1(or 3),2-species (32.6%), and triacylglycerol (rapeseed oil) at the rate of 3ml/h for 1 h through a cannula inserted into the stomach. The lymph fluids were collected every hour for 5 h after starting the infusion of the lipid emulsions, and the lymph chylomicrons were isolated, purified and analyzed. Test emulsions were prepared to provide the same amount of fatty acids (144mg/h) as that in these acylglycerols. The rates for triacylglycerol transport at 2–3 h and for cholesterol transport by chylomicrons at 2–3 h and 3–4 h of the experimental period in rats infused with the diacylglycerol emulsion were significantly lower than the corresponding values in the rats infused with the triacylglycerol emulsion. As a consequence, the cumulative value for triacylglycerol transport at the end of the experimental period in rats infused with the diacylglycerol emulsion was significantly lower than the corresponding value in the rats infused with the triacylglycerol emulsion. In addition, cumulative values for cholesterol transport from 3h to the end of the experimental period were significantly lower in the former than in the latter. There was no difference in the total fatty acid compositon of chylomicron-triacylglycerol between the rats receiving the triacylglycerol and diacylglycerol emulsions. However, a considerable difference existed in the fatty acid composition at the 2-position of triacylglycerol between the two groups of rats. Thus, intragastric infusion of diacylglycerol mainly consisting of the 1,3-species compared to triacylglycerol not only altered the rate of lipid transport by lymph chylomicrons but also altered the structure of the triacylglycerol moiety in the rats.  相似文献   

15.
Chylomicron retention disease is characterized by fat malabsorption, hypocholesterolemia, normal fasting triglycerides, and marked intestinal steatosis despite the presence of both plasma and intestinal apoprotein B. The defect remains unknown but presumably involves the synthesis or secretion of chylomicrons. The present investigation examines this hypothesis by studying the biosynthesis of chylomicrons in cultured jejunal explants and by defining the quantitative and qualitative abnormalities of plasma lipids and of circulating lipoproteins. Following 2-3 years of a low fat diet supplemented with medium chain triglycerides, six patients with chylomicron retention disease had significantly higher triglyceride (TG) levels coupled with a decrease in both free (FC) and esterified cholesterol (EC) as well as in essential fatty acids and phospholipids (PL) when compared to healthy controls. The low total plasma cholesterol was largely accounted for by low levels of both low density (LDL) and high density lipoprotein (HDL) cholesterol. VLDL and LDL were characterized by a diminished percentage of CE with an increase of TG while HDL contained relatively more FC as well as PL and less CE. The diameter of VLDL was larger whereas those of LDL and HDL were smaller than in normal controls. Jejunal explants, when incubated with [14C]palmitate, were capable of normal biosynthesis of TG, diglycerides, PL, and CE. These lipids, however, except for PL, were retained in the tissue and could not be secreted into the culture medium. Incubation of intestinal biopsies with [3H]leucine and [14C]mannose resulted in normal protein synthesis and reduced glycosylation. The presence of intestinal apoB-48 was confirmed by immunoblot using 2D8 antibodies. These data suggest that the intestinal defect in this disease results from a disorder of the final assembly of chylomicrons or in the mechanism of their exocytosis.  相似文献   

16.
Peroxisomes were isolated from liver tissue of control and clofibrate-treated adult male NMRI mice and Sprague-Dawley rats. Phospholipids, cholesterol, triglycerides and free fatty acids were measured in the peroxisomes. The fatty acid profiles of the phosphatidylethanolamine, the phosphatidylcholine, the triglyceride and the free fatty acid fractions were also analyzed. Phosphatidylethanolamine was the dominating phospholipid in peroxisomes from untreated animals. The fatty acid profiles of phosphatidylethanolamine, free fatty acids and triglycerides were similar for untreated mice and rats but differences between the species were observed in the pattern derived from phosphatidylcholine. Phosphatidylcholine was the most abundant phospholipid after clofibrate treatment. Clofibrate treatment caused an increase in the concentrations of phospholipids and unsaturated long-chain fatty acids and a decrease in the concentrations of triglycerides, free fatty acids, cholesterol and shorter saturated fatty acids.  相似文献   

17.
Isolated livers perfused with washed corn oil chylomicrons labeled in vivo with palmitic acid-1-(14)C removed a large proportion of the chylomicrons. Slices from these livers oxidized chylomicron fatty acid esters to both carbon dioxide and acetoacetate. The liver slices also generated free fatty acids from chylomicron lipids and converted chylomicron triglycerides to phospholipids. Similar activities were observed in rat liver slices prepared shortly after the intravenous administration of chylomicrons to intact rats. The observed chylomicron uptake and lipid conversions were similar in livers from both fed and fasted rats. Fasting increased the oxidation of chylomicron fatty acid esters by livers labeled in vivo and by perfusion. In livers removed from intact rats given labeled chylomicrons, the triglyceride-(14)C to phospholipid-(14)C ratio was high, a finding unexpected if the liver had acquired this (14)C by removal of circulating fatty acids formed by extrahepatic lipolysis. These results demonstrate the ability of the liver to remove and utilize chylomicrons directly and suggest that direct removal accounts for a significant portion of the chylomicron fatty acids utilized by the liver of intact rats.  相似文献   

18.
Changes in the plasma membrane lipid composition have been related to a decrease in sperm quality during cryopreservation. Antifreeze proteins (AFPs) have been tested in different species because of their ability to depress the freezing point and their potential interaction with membranes, but controversial effects were reported. In the present study we analyzed separately the lipid composition of two sperm membrane domains, head plasma membrane (HM) and flagellar membrane (FM), after cryopreservation with an extender containing 5% dimethyl sulfoxide (DMSO) either alone or with AFPI or AFPIII (1 μg/ml). We used sperm from a teleost, Sparus aurata, because the lack of acrosome avoids changes of lipid profiles due to capacitation process or acrosomal losses during freezing/thawing. Comparing with the control (cryopreservation with 5% DMSO alone), the addition of AFPIII increased the velocity, linearity of movement, and percentage of viable cells. In addition, freezing with DMSO alone increased the phosphatidyl-serine content as well as the saturated fatty acids and decreased the unsaturated ones (mainly polyunsaturated) both in HM and FM. These changes in the lipid components were highly avoided with the addition of AFPIII. HM had a higher amount of saturated fatty acids than FM and was more affected by cryopreservation without AFPs. The percentage of viable cells was positively correlated with the amount of unsaturated fatty acids in the HM, whereas the motility parameters were positively correlated with both FM and HM amount of unsaturated fatty acids. AFPs, especially AFPIII, seem to have interacted with unsaturated fatty acids, stabilizing the plasma membrane organization during cryopreservation and contributing to improve sperm quality after thawing.  相似文献   

19.
AIMS: Isolation and characterization of unsaturated fatty acids during bacteriorhodopsin preparation from Halobacterium halobium. METHODS AND RESULTS: Halobacterium halobium was cultivated in a composite medium. Cells were collected by centrifugation followed by ultrasonic disruption, and the resulting suspension was subject to centrifugation for preparation of both pellet and supernatant. The pellet was saved in order to prepare bacteriorhodopsin, while the supernatant was used for the isolation of crude fatty acids by saponification and extraction. Crystallization then took place in acetone at -16 degrees C to remove fatty acids in which the carbon chain length was shorter than 13. The sample was obtained after purification and analysed by gas chromatography. The results demonstrated that Halobacterium halobium could synthesize multiple unsaturated fatty acids, particularly the three important polyunsaturated fatty acids arachidonic acid (1.12%), eicosapentaenoic acid (16.76%) and docosahexaenoic acid (9.38%). CONCLUSION: Important unsaturated fatty acids were isolated and characterized from the waste, which was produced during the preparation of bacteriorhodopsin from Halobacterium halobium. SIGNIFICANCE AND IMPACT OF THE STUDY: Halobacterium halobium has already been used for decades to prepare bacteriorhodopsin. We found that several important unsaturated fatty acids could be extracted from the bacterial waste, which extends its application scope and might bring additional benefits to humanity.  相似文献   

20.
在适宜浓度(0.1~0.4m mol/L)的胆固醇存在下,经冰冻处理的小麦叶肉原生质体在化冻后存活率均高于对照,其中以0.4m mol/L胆固醇对原生质体的保护效果最佳。胆固醇对卷心菜叶肉原生质体的抗冰冻能力具有类似效果。从而表明外源胆固醇与质膜结合后,对稳定质膜结构和保持正常功能具有一定的作用。  相似文献   

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