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1.
目的:用MTT法检测树突状细胞(DC)杀伤肿瘤细胞的活性.方法:将人外周血分离出单个核细胞,在细胞因子的刺激下形成DC,观察其细胞形态、增殖情况,采用MTT法检测DC对肿瘤K562细胞的杀伤活性.结果:DC培养至第1l天时,增殖高达120倍,经MTT法检测证实,DC对K562细胞的4h杀伤活性为3l%.结论:MTT法检测进一步证实了DC对肿瘤细胞有较强的细胞毒作用.  相似文献   

2.
目的:比较不同来源盐酸多巴酚丁胺注射液体外细胞毒性的差别,为遴选优质药品、保障用药安全提供科学依据.方法:将不同来源国产盐酸多巴酚丁胺注射液原液以及稀释液,与L929细胞接触培养,通过倒置相差显微镜观察其形态,采用MTT法量化细胞毒性,计算相对增值率和IC50值,进行细胞毒性评价.并将国产盐酸多巴酚丁胺注射液与进口制剂进行比较.结果:8个厂家生产的盐酸多巴酚丁胺注射液原液及进口产品的细胞毒性分级均为4级.不同企业生产的盐酸多巴酚丁胺注射液的IC50值差异较大.盐酸多巴酚丁胺注射液C和F的IC50值较小,说明这两个企业产品的细胞毒性较其他样品大.进口盐酸多巴酚丁胺注射液的IC50值为99.9 mg·L-1,较其他样品高,说明其细胞毒性较其他样品低.进口盐酸多巴酚丁胺注射液中仅添加亚硫酸氢钠一种辅料,国产盐酸多巴酚丁胺注射液中添加的辅料主要为依地酸二钠、亚硫酸氢钠、盐酸半胱氨酸等.按各辅料在国产盐酸多巴酚丁酸注射液中的浓度测其细胞毒性,依地酸二钠、盐酸半胱氨酸和亚硫酸氢钠的细胞毒性分级均为3级.将依地酸二钠的浓度稀释为0.03 mg·ml-时,其细胞毒性分级为1级,无细胞毒性.将盐酸半胱氨酸稀释到0.11 mg·ml-1时,其细胞毒性分级为1级,基本无细胞毒性.结论:不同厂家生产的盐酸多巴酚丁胺注射液由于生产条件不同、处方不同等原因,这些产品的细胞毒性存在较大的差别.与进口制剂比较,国产制剂中添加的辅料浓度较大,可适当降低其浓度,以减少药物制剂的细胞毒性,从而提高药品的安全性.  相似文献   

3.
Dierickx PJ 《Toxicology》2000,150(1-3):159-169
The cytotoxicity of the MEIC reference chemicals was investigated in rat hepatoma-derived Fa32 cells. The total protein content was measured as an endpoint after exposure times of 30 min and 24 h, both in normal and glutathione-depleted cells. The neutral red uptake inhibition and the MTT conversion were also measured after 30 min. On average, the cytotoxicity was higher in glutathione-depleted cells when compared to normal cells, and was lower after 30 min than after 24 h. Evidence was obtained for lysosomal attack (of five chemicals) or mitochondrial dysfunction (of six chemicals) as the primary intoxication mechanism. Malathion and mercuric chloride belong to both series of chemicals. Good to excellent correlations were observed when the 50% inhibitory concentrations of the six different in vitro assays were compared. When the six in vitro assays in Fa32 cells were compared with the human toxicity, the correlation coefficient was almost always identical to that obtained previously in human hepatoma-derived Hep G2 cells. The latter was the best acute in vitro assay for the prediction of human toxicity within the MEIC study. Altogether the results integrate very well with the basal cytotoxicity concept (Ekwall, B., 1995. The basal cytotoxicity concept. In: Goldberg, A.M., Van Zutphen, L.F.M. (Eds.), The World Congress on Alternatives and Animal Use in the Life Sciences: Education, Research, Testing. Mary Ann Liebert Publishers, New York, pp. 721–725).  相似文献   

4.
顺铂和卡铂对胃癌细胞系的毒性作用比较   总被引:1,自引:0,他引:1  
卜书红  李方  王晶  杨传敏 《医药导报》2001,20(10):621-621
目的:体外比较顺铂和卡铂对胃癌细胞系的毒性作用,为临床化疗提供参考。方法:采用二甲基噻唑基四氮唑试验(MTT)在不同的培养时间,用不同浓度的药物,通过其细胞抑制率和半数抑制率(IC50)比较两种药物的细胞毒性。结果:不同培养时间的肿瘤细胞抑制率顺铂明显高于卡铂(P<0.05),顺铂的IC50也远小于卡铂。结论:顺铂对胃癌细胞系的体外细胞毒性作用大于卡铂。  相似文献   

5.
Ginkgolic acids and related alkylphenols (e.g. cardanols and cardols) have been recognized as hazardous compounds with suspected cytotoxic, allergenic, mutagenic and carcinogenic properties. To determine whether the phase I metabolism could contribute to their cytotoxicity, we investigated the cytotoxicity of one model compound, ginkgolic acid (15:1), using in vitro bioassay systems. In the first step, cytochrome P450 enzymes involved in ginkgolic acid metabolism were investigated in rat liver microsomes; then, two in vitro cell-based assay systems, primary rat hepatocytes and HepG2 cells, were used to study and the measurement of MTT reduction was used to assess cell viability. Results indicated that the cytotoxicity of ginkgolic acid in primary rat hepatocytes was lower than in HepG2 cells. Ginkgolic acid was demonstrated less cytotoxicity in four-day-cultured primary rat hepatocytes than in 20-h cultured ones. Co-incubation with selective CYP inhibitors, α-naphthoflavone and ketoconazole, could decrease the cytotoxicity of ginkgolic acid in primary rat hepatocytes. In agreement, pretreatment with selective CYP inducers, β-naphthoflavone and rifampin, could increase the cytotoxicity of ginkgolic acid in HepG2 cells. These findings suggest that HepG2 cells were more sensitive to the cytotoxicity of ginkgolic acid than primary rat hepatocytes, and CYP1A and CYP3A could metabolize ginkgolic acid to more toxic compounds.  相似文献   

6.
N-三甲基壳聚糖包衣脂质体的细胞毒性考察   总被引:1,自引:1,他引:0       下载免费PDF全文
何文  张小杰 《中国药师》2010,13(10):1416-1419
目的:评价不同季铵化程度的N-三甲基壳聚糖(TMC)包衣的空白脂质体对小鼠成纤维细胞L-929的毒性。方法:采用四步合成法合成不同季铵化程度的TMC,并采用IR对其结构进行确证和表征,^1H—NMR确定其季铵化程度。采用MTT比色法评价TMC作为阳离子脂质体包衣材料,在不同浓度和不同季铵化程度时对小鼠成纤维细胞L-929的毒性,计算相对增殖率,并参考《美国药典》的评价标准对其进行毒性分级。结果:IR分析TMC的特征吸收峰已明显发生了改变,表明TMC已经成功合成,^1H-NMR分析所合成的TMC,季铵化程度分别为20.2%、40.1%、64.4%,不同季铵化的TMC在浓度为0.02%,0.05%(w/w),细胞增殖率均大于80%,TMC20在浓度增大到0.2%(w/w)时,细胞增殖率仍高于80%,细胞毒性为1级,即无细胞毒性,但随着浓度和季铵化程度的提高,细胞毒性均相应增大。结论:所合成的TMC在一定的浓度和季铵化范围内,细胞毒性较低,又鉴于其具有良好的生物降解性、亲水性和生物粘附性,可望作为阳离子脂质体理想的包衣材料。  相似文献   

7.
Fumonisin B1 (FB1), the most prevalent member of toxins produced by several species of Fusarium molds, which occur mainly in maize, causes several fatal hepatopathies and nephropathies of animals. The current study was scrutinized to ascertain different cytotoxic and morphological transformations in rat hepatocytes induced by the treatments of diverse concentrations (300, 500, or 1000 μM) of fumonisin B1 in vitro, using both monolayer and spheroid cultures. In each hepatocyte culture, the cytotoxicity of FB1 was augmented in dose- and time-response manners. Morphological transformations among FB1-treated groups integrated accumulation of lipid droplets, cytoplasmic vacuolation in hepatocyte monolayers, and bleb formation in the hepatocyte spheroids. Additionally, electron microscopy revealed the loss of microvilli, mitochondrial swelling, and formation of lamellar membranous whorl in the vacuoles and bile canaliculi-like structures. Appearance of electron dense bodies in the monolayers, and loss of cell-to-cell contact in spheroids were depicted in 1000 μM FB1-treated hepatocytes. These outcomes insinuate different vital events in explaining morphological transformations in the cell membrane and organelles, induced by fumonisins in rat hepatocytes.  相似文献   

8.
ABSTRACT

In this study, branchlets, fruit, or bark of Taxus baccata. L. as well as branchlets or fruits of two other species of Iranian conifers, namely, Platycladus orientalis. France and Cupressus sempervirens. L. var. horizentalis. (Mill) Gunde were collected, identified, and the cytotoxic effects of hydroalcoholic extracts on three human tumor cell lines were determined. Different concentrations of extracts were added to cultured cells and incubated for 72 h. Cell survival was evaluated using the MTT assay. Extracts from bark of female Taxus baccata. showed inhibitory activities against Hela cells. The extracts of the branchlets of male and female T. baccata. as well as obtained extract from fruits of P. orientalis. showed inhibitory activities against MDA-MB-468 cells, whereas the extracts of branchlets of female T. baccata. showed inhibitory activities against KB cells. In conclusion, obtained extract from bark of T. baccata. showed comparable cytotoxic effect to doxorubicin against Hela cells.  相似文献   

9.
Capsaicin is a natural product of Capsicum peppers, excitatory effects of which have been shown to be mediated by the recently cloned vanilloid receptor 1 (VR1). Since previous studies have shown that capsaicin inhibits protein synthesis, experiments were performed to investigate whether this effect is mediated by VR1 receptor on cultured monkey kidney cells (Vero cells). The capsaicin uptake was assessed in cellular homogenate and in medium by high-performance liquid chromatography (HPLC) separation and quantification on C18 reverse-phase column and fluorescence detection. Toxic effects were assessed by incorporation of [3H]L-leucine into cellular proteins in the presence of capsazepine, the VR1 vanilloid receptor antagonist and Ruthenium red or tyrosine or calcium. Capsazepine (1 to 256 μM) did not modify the uptake rate of capsaicin for incubation times up to 24 h and did not antagonize capsaicin-induced protein synthesis inhibition. It rather inhibited protein synthesis per se from 100 to 256 μM. Ruthenium red which blocks mitochondrial calcium uptake, inhibited protein synthesis and did not antagonise or increase synergistically the effects of capsaicin. Interestingly in a medium deprived of calcium and supplemented by calcium chloride (10–50 μM) the protein synthesis inhibition induced by capsaicin is antagonised somehow. There was no prevention of capsaicin diffusion into the cells. Tyrosine, which seems to be the best preventive agent of capsaicin inhibitory effects, prevents its metabolism but not its diffusion. Capsaicin might enter cells by diffusion and interfere with protein synthesis machinery by competition with tyrosine which in turn prevents the metabolism of capsaicin. The results of the present study suggest that cell responses to capsaicin may be transduced through at least two molecular pathways, one involving VR1, since the receptor antagonist capsazepine fails to prevent the inhibitory effect of capsaicin in Vero cells of renal origin. Received: 27 August 1999 / Accepted: 3 November 1999  相似文献   

10.
Natural killer (NK) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. Because bone marrow-derived hematopoietic stem cells (HSCs), lymphoid protenitors, can give rise to NK cells, NK ontogeny has been considered to be exclusively lymphoid. Here, we show that porcine c-kit+ bone marrow cells (c-kit+ BM cells) develop into NK cells in vitro in the presence of various cytokines [interleukin (IL)-2, IL-7, IL-15, IL-21, stem cell factor (SCF), and fms-like tyrosine kinase-3 ligand (FLT3L)]. Adding hydrocortisone (HDC) and stromal cells greatly increases the frequency of c-kit+ BM cells that give rise to CD2+CD8+ NK cells. Also, intracellular levels of perforin, granzyme B, and NKG2D were determined by RT-PCR and western blotting analysis. It was found that of perforin, granzyme B, and NKG2D levels significantly were increased in cytokine-stimulated c-kit+ BM cells than those of controls. And, we compared the ability of the cytotoxicity of CD2+CD8+ NK cells differentiated by cytokines from c-kit+ BM cells against K562 target cells for 28 days. Cytokines-induced NK cells as effector cells were incubated with K562 cells as target in a ratio of 100:1 for 4 h once a week. In results, CD2+CD8+ NK cells induced by cytokines and stromal cells showed a significantly increased cytotoxicity 21 days later. Whereas, our results indicated that c-kit+ BM cells not pretreated with cytokines have lower levels of cytotoxicity. Taken together, this study suggests that cytokines-induced NK cells from porcine c-kit+ BM cells may be used as adoptive transfer therapy if the known obstacles to xenografting (e.g. immune and non-immune problems) were overcome in the future.  相似文献   

11.
目的采用星点设计优化处方,制备三氧化二砷固体脂质纳米粒并探讨其对4种肿瘤细胞的抑制作用。方法采用乳化超声法制备三氧化二砷固体脂质纳米粒。采用MTT法考察三氧化二砷固体脂质纳米粒对4种肿瘤细胞的体外抑制作用。结果筛选的最优处方为PEG-单硬脂酸甘油酯用量0.11 g,大豆卵磷脂的用量0.18 g。平均粒径为131.54 nm,包封率73.46%,载药量为1.07%。不同浓度三氧化二砷固体脂质纳米粒对4种细胞均有抑制增殖作用。结论采用乳化超声法制得的三氧化二砷固体脂质纳米粒具有较好的稳定性,粒子分布均匀,符合制剂学要求。携带阳离子三氧化二砷固体脂质纳米粒组抑制作用最强。  相似文献   

12.
摘 要 目的:考察不同型号的普朗尼克(pluronic)对人乳腺癌细胞(MCF 7)及对多西他赛耐药的人乳腺癌细胞(MCF 7/DTX)毒性及摄取的影响,筛选出最优型号,以供后续制备普朗尼克载多西他赛胶束。方法: 选取L61、P85、P105、P123、F68、F127共6种型号普朗尼克,采用CCK 8法测定其对MCF 7及MCF 7/DTX的细胞毒性,高效液相色谱(HPLC)法测定两种细胞对多西他赛的摄取量。结果: F68、F127对MCF 7及MCF 7/DTX的毒性较弱,F127的肿瘤细胞毒性最弱;而L61、P85、P105、P123对两种细胞的毒性相对较强,且L61的肿瘤细胞毒性最强。在浓度为50~1 000 μg·ml-1的范围内,普朗尼克的肿瘤细胞毒性与其浓度成正相关,浓度越高,毒性越强。在24~72 h内,同一浓度的普朗尼克,作用时间越长,毒性越强,细胞毒性与作用时间也成正相关。6种型号普朗尼克均可促进MCF 7对多西他赛的摄取,仅有P85、P105、P123可促进MCF 7/DTX的摄取,而L61、F68、F127对耐药细胞摄取量基本无促进作用,其中P123促进两种肿瘤细胞摄取多西他赛的能力最强。结论: HLB值小的普朗尼克,肿瘤细胞毒性更强。随着浓度升高,作用时间增长,其毒性增强。P85、P105、P123可促进耐药细胞对多西他赛的摄取。  相似文献   

13.
 The cytotoxic effects of propyl gallate (PG), its related gallates and gallic acid have been studied in freshly isolated rat hepatocytes. Addition of PG (0.5–2.0 mM) to hepatocyte suspension elicited concentration-dependent cell death accompanied by losses of intracellular ATP, adenine nucleotide pools, glutathione (GSH) and protein thiols. The rapid loss of intracellular ATP preceded the onset of cell death caused by PG. In the comparative toxic effects of PG and related gallates at concentration of 1 mM, octyl gallate (OG), dodecyl gallate (DG) and butyl gallate (BG) elicited an abrupt depletion of ATP, followed by an acute cell death. These gallates were more toxic than PG; the toxic effects of PG were similar to those of methyl gallate (MG) and ethyl gallate (EG). In mitochondria isolated from rat liver, PG caused a concentration-dependent increase in the rate of state 4 oxygen consumption, indicating an uncoupling effect. The rate of state 3 oxygen consumption was inhibited by OG and DG. According to the respiratory control index, the order of impairment potency to mitochondria was OG>BG, DG>PG>EG, MG>gallic acid. These results indicate that PG and related gallates are toxic to hepatocytes and that the acute cytotoxicity may be due to mitochondrial dysfunction. Received: 16 May 1994 / Accepted: 15 August 1994  相似文献   

14.
Context: Termitomyces clypeatus (Lyophyllaceae) is a filamentous edible mushroom, having ethnomedicinal uses. However, information about the antioxidant, anticancer and antitumour properties of this mushroom remains to be elucidated.

Objective: The study examines the in vitro antioxidant, anticancer and in vivo antitumour activity of T. clypeatus.

Materials and methods: Antioxidant activity was evaluated with seven in vitro assays. Cytotoxicity of T. clypeatus was tested against a panel of cancer cells lines including U373MG, MDA-MB-468, HepG2, HL-60, A549, U937, OAW-42 and Y-79 using MTT assay. The antitumour activity of aqueous extract was evaluated against Ehrlich ascites carcinoma (EAC) tumour model in Swiss albino mice.

Results: HPLC analysis of aqueous extract revealed the presence of sugar entities. Termitomyces clypeatus showed excellent in vitro antioxidant activity. Termitomyces clypeatus was found cytotoxic against all cancer cells, among which it showed higher activity against U937 (IC50 25?±?1.02?μg/mL). Treatment of EAC-bearing mice with varied doses of aqueous extract significantly (p?<?0.01) reduced tumour volume, viable tumour cell count and improved haemoglobin content, RBC count, mean survival time, tumour inhibition and % increase life span. The enhanced antioxidant status in treated animals was evident from the decline in the levels of lipid peroxidation, increased levels of glutathione, catalase and superoxide dismutase.

Discussion: The analyzed data indicate that the aqueous extract of T. clypeatus exhibits significant antitumour activity, which might be due to the antioxidant effects on EAC bearing hosts.

Conclusion: Termitomyces clypeatus possesses anticancer activity, valuable for application in food and drug products.  相似文献   

15.
A number of cytotoxicity assays are currently available, each of them using specific approach to detect different aspects of cell viability, such as cell integrity, proliferation and metabolic functions. In this study we compared the potential of five commonly employed cytotoxicity assays (WST-1, XTT, MTT, Brilliant blue and Neutral red assay) to detect antiproliferative effects of three selenium compounds, sodium selenite, seleno-l-methionine (SeMet) and Se-(Methyl)selenocysteine (SeMCys) on three colorectal cancer cell lines in vitro. Cells were exposed to the selected selenium compounds in the concentration range of 0–256 μM during 48 h. WST-1 and XTT failed to detect cytotoxic effect, with the exception of the highest concentration of selenium compounds tested. Conversely, the metabolic activity of selenium treated cells measured by WST-1 and XTT significantly increased in comparison to untreated controls. MTT, Neutral red and Brilliant blue assays were more sensitive and yielded mutually comparable results, with significant decrease of measured parameters in a concentration-dependent manner. To a smaller extent, the results were affected by the different chemical nature of the selenium compounds tested as well as by the biological properties of individual cell lines.  相似文献   

16.
Primary cultures or established cell lines of vertebrates are commonly used to analyse the mutagenic, embryotoxic or teratogenic potential of environmental factors, drugs and xenobiotics in vitro. However, these cellular systems do not include developmental processes from early embryonic stages up to terminally differentiated cell types. An alternative approach has been offered by permanent lines of pluripotent stem cells of embryonic origin, such as embryonic carcinoma (EC), embryonic stem (ES) and embryonic germ (EG) cells. The undifferentiated stem cell lines are characterized by nearly unlimited self-renewal capacity and have been shown to differentiate in vitro into cells of all three primary germ layers. Pluripotent embryonic stem cell lines recapitulate cellular developmental processes and gene expression patterns of early embryogenesis during in vitro differentiation, data which are summarized in this review. In addition, recent studies are presented which investigated mutagenic, cytotoxic and embryotoxic effects of chemical substances using in vitro systems of pluripotent embryonic stem cells. Furthermore, an outlook is given on future molecular technologies using embryonic stem cells in developmental toxicology and embryotoxicology.  相似文献   

17.
1. The present study investigated the use of drugs that affect calcium (Ca2 +) levels and thus reduction of triamcinolone (TA)‐induced cytotoxicity on human retinal epithelial (ARPE19) cells. 2. Four groups were compared: ARPE19 cells alone, cells exposed to TA (0.1 mg/mL), cells that have been pretreated with one of the testing agents for 30 min before the addition of TA, and cells that have only been treated with one of the testing agents. Pinacidil (PIN) and its analogue, P1060, were used to test the effect of potassium (K+) channel opening on TA‐induced toxicity. Verapamil (VP) and diltiazem (DZ) were used to test their Ca2 + channel blocking effect. The cell viability under different settings was assessed using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay. Ca2 +‐imaging was used to determine the changes in intracellular Ca2 + levels [(Ca2 +)i] upon different treatments. 3. Both PIN and P1060 reduced TA‐induced toxicity. Verapamil and DZ increased the viability of cells treated with TA significantly, suggesting that the excessive influx of Ca2 + was one of the main contributory factors to the TA‐induced toxicity. 4. The results suggest that the prevention of Ca2 + entry may be effective in the reduction of cell necrosis in the presence of TA.  相似文献   

18.
人工蛹虫草子实体提取物对肾小球系膜细胞增殖的影响   总被引:1,自引:1,他引:1  
李维亮  裘军  杨楠  阮金兰 《中国药师》2008,11(5):501-503
目的:探讨人工蛹虫草不同溶剂提取部位对大鼠肾小球系膜细胞增殖的影响.方法:系膜细胞与不同溶剂提取物共同培养或加入LPS刺激细胞增殖,培养68 h后,采用四甲基偶氮唑蓝(Mar)法观察人工蛹虫草不同溶剂提取物对培养的肾小球系膜细胞增殖的影响.结果:正丁醇提取物在2-250μg·ml-1浓度范围内呈浓度依赖性抑制系膜细胞增殖;乙酸乙酯提取物的抑制浓度为0.51~100μg·ml-1,甲醇提取物的抑制浓度为62.5~250.0μg·ml-1,而水提醇沉提取物在0.5~12.5μg·ml-1范围内具有促进系膜细胞增殖作用;乙醇提取物作用不明显.结论:人工蛹虫草子实体存在对肾小球系膜细胞增殖有抑制和促进作用的成分.  相似文献   

19.
The main objective of this study was to assess an in-house 3T3 NRU cytotoxicity assay for compatibility with a prediction model for acute rodent oral toxicity endorsed by an NIEHS–ICCVAM workshop. The aim is to use the NRU assay as one test component of HTS strategies for both acute oral toxicity and acute skin irritation, enabling the rejection of the most toxic materials and prioritisation of other materials for further testing. Groups of model cytotoxins and irritants were tested using the NRU assay and their EC50 values obtained from dose–response curves. These values were compared with those estimated from a limited (three)-dose protocol, deemed more suitable for HTS. A good correlation was observed between the EC50 values from both dose–response curves (R2=0.94). The relationships between EC50 values and acute rodent oral toxicity were compared by application of the prediction model to the model cytotoxins. The results from both full and limited dose–responses fitted within the acceptance limits of the prediction model, with regression lines similar to that of the model. Results indicated that the performance of the currently used 3T3 NRU cytotoxicity assay was similar to that of the assays used to generate the data employed in developing the prediction model. This prediction model can be applied with both the standard and HT assays to estimate acute rodent oral toxicity.  相似文献   

20.
目的:对不同来源一次性使用无菌注射器用活塞体外细胞毒性进行评价,为科学监管提供技术支持。方法:将一次性使用无菌注射器用活塞浸提液分别与L929细胞接触培养,采用四唑盐比色法(MTT法)量化细胞毒性,计算相对增殖率,并进行细胞毒性评价。结果:不同厂家相同浸提比例,产品之间细胞毒性均存在差异,同一厂家不同浸提比例的条件下,产品细胞毒性未发现明显变化。结论:不同厂家产品配方及生产工艺不尽相同可能是导致不同厂家的产品的细胞毒性存在差异的原因,建议在产品标准中生物性能技术要求中增加细胞毒性评价项。  相似文献   

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