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1.
目的 探讨内皮素(ET -1)对活化的肝星状细胞(HSCs)细胞外信号调节激酶 1(ERK1)表达、细胞周期及细胞内Ca2 影响。方法 采用Western印迹观察不同浓度ET -1对HSCsERK1的表达,采用流式细胞术观察ET -1对细胞周期的影响,并借助激光共聚焦显微镜技术观察ET- 1对HSCsCa2 浓度变化和细胞面积的影响。结果 ET -1可剂量依赖性地抑制HSCsERK1的表达,最高浓度ET -1可显著抑制ERK1表达,同对照组相比下降(10 .91±3.36 ) % (P <0 .0 5 ) ,还可抑制HSCsDNA合成,使增殖指数由(32 .6 3±0 .36 ) %下降至(2 4 .72±0 .12 ) % (P <0 .0 1) ,同时ET- 1可引起HSCs [Ca2 ]i的升高及细胞面积的减少(P <0 .0 5 )。结论 ET- 1对HSCs发挥着促进HSCs收缩与抑制其增殖的双重作用,所以应用药物治疗慢性肝病既要注意抑制ET的收缩效应同时又要兼顾对其本身具有的抑制增殖的双重作用。  相似文献   

2.
目的:探讨心力衰竭(心衰)家兔心肌细胞钙释放异常与收缩功能的关系。方法:22只家兔随机分为2组:假手术组(n=11)和心衰组(n=11);通过超容量负荷联合压力负荷制备家兔心衰模型,利用心脏多普勒观察手术前后家兔心功能的变化;采用常规酶解法分离心室肌细胞,经Fluo-3/AM负载后,利用激光共聚焦显微技术,观察咖啡因诱发钙瞬变过程中细胞内钙浓度的动态变化;采用Western blot法测定心肌组织L型钙通道α1亚单位(LTCCα1)、肌浆网钙释放通道(RyR2)的表达水平。结果:与假手术组相比,心衰组家兔射血分数明显降低(0.46±0.03 vs 0.73±0.05,P<0.01);咖啡因诱发的钙瞬变幅度明显下降(16.00±3.54 FI vs 43.50±6.22 FI,P<0.01)、峰值到达时间明显延长(129.8±14.5 s vs 52.2±7.4 s, P<0.01);LTCCα1(RLTCC-α1/actin:0.287±0.029 vs 0.624±0.009,P<0.01)和RyR2表达水平明显减少(RRyR2/actin:0.106±0.001 vs.0.203±0.011,P<0.01),均有极显著性差异。结论:LTCCα1亚单位和RyR2表达减少导致钙释放异常是心衰心肌收缩力降低的机制之一。  相似文献   

3.
槟榔碱促结肠平滑肌细胞收缩及对胞内钙离子浓度的影响   总被引:3,自引:2,他引:3  
目的:观察氢溴酸槟榔碱(Ah)促进培养的结肠平滑肌细胞收缩作用及对胞内游离Ca2 浓度的影响.方法:培养大鼠结肠平滑肌细胞,分为4组:正常对照组、Ah刺激组、乙酰胆碱(Ach)刺激组、阿托品预处理组.应用特异性Ca2 荧光批示剂Fluo-3/AM负载细胞,激光共聚焦显微镜检测游离Ca2 浓度和细胞收缩率.结果:正常对照组细胞未发生自主性收缩,因指示剂的衰减,荧光强度(FI)有递减的趋势; Ah刺激组在加药后短时间内胞内钙离子浓度迅速升高,出现一个波峰,FI平均基础值与峰值有差异(P<0.05),而后胞内钙离子浓度再缓慢攀升,在484.0±47.6 s达到高峰,而后有一个较长时间的平台期,在1 400 s左右恢复至静息水平,FI基础值与峰值有显著差异 (P<0.01),细胞收缩百分数为20.70%±0.07%; Ach刺激组在加药后胞内钙离子浓度升高,形成一个小波峰,FI基础值和峰值差异有显著性(P<0.01).随后胞内钙离子浓度缓慢攀升, 在600 s左右达到高峰,而后有一个较长时间的平台期.FI基础值与峰值差异有显著性 (P<0.001);经阿托品预孵育处理后的细胞,加入Ah后,其收缩效应被完全抑制.结论:Ah可引起结肠平滑肌细胞收缩,升高细胞内游离Ca2 浓度.其收缩效应可以被M受体阻断剂阿托品所抑制.  相似文献   

4.
目的:探讨活血化瘀中药益肝康、丹参小复方、丹参对IL-1β刺激的活化的大鼠肝星状细胞(HSCs)增殖及基质金属蛋白酶抑制因子 mRNA(TIMP mRNA)表达的影响.方法:体外培养活化的大鼠HSC,随机分为8 组:对照组(A组)、IL-1β 10 μg/L(B组)、IL- 1β 10μg/L 益肝康2 g/L干预组(C组)、IL- 1β 10 μg/L 丹参小复方2g/L干预组(D组)、 IL-1β 10μg/L 丹参2g/L干预组(E组)、丹参 2 g/L(F组)、丹参小复方2 g/L(G组)、益肝康 2 g/L(H组).加药后24 h.应用活细胞计数试剂盒-CCK-8检测各组HSC增殖,采用半定量 RT-PCR方法检测各组HSC TIMP-1 mRNA的表达.结果:A组HSC增殖和TIMP-1 mRNA表达强于F、G、H组(1.291±0.09 vs 1.055±0.105, 1±0.07,0.883±0.06,P<0.01:0.591±0.064 vs 0.493±0.088.0.458±0.076.0.356±0.046. P<0.05或P<0.01);H组HSC增殖和TIMP-1 mRNA表达低于F组和G组(P<0.05);B组HSC 增殖和TIMP-1 mRNA表达均明显强于A组 (1.575±0.017 vs 1.291±0,09,P<0.01;1.369± 0.097 vs 0.591±0.064,P<0.01)和C、D、E组 (1.575±0.017 vs 0.906±0.09,1.015±0.081, 1.097±0.038,P<0.01;1.369±0.097 vs 0.694 ±0.078,0.854±0.05,0.898±0.12,P<0.01);C 组HSC增殖和TIMP-1 mRNA表达低于D组和 E组(P<0.05).结论:益肝康等活血化瘀中药能抑制IL-1β刺激的HSCs增殖及TIMP-1 mRNA表达,发挥其抗肝纤维化功效.益肝康抗肝纤维化作用强于丹参小复方和丹参单药.  相似文献   

5.
目的探讨营养性低钙与慢性氟中毒细胞内钙超载的相关性。方法采用饮水加入氟化钠进行大鼠染毒实验,配以低钙饮食,应用Fura-2/AM荧光指示剂标记单细胞,显微荧光标记测定慢性氟中毒大鼠肝、肾、脑细胞内游离钙([Ca2 ]i)的变化。结果氟可刺激细胞内[Ca2 ]i水平增高,正常饮食投氟组肝、肾、脑细胞内[Ca2 ]i水平分别为(124.11±11.35)、(127.20±14.40)、(136.75±15.91)nmol/L,与正常饮食对照组(86.37±27.55)、(95.65±8.75)、(99.20±12.89)nmol/L比较,差异有统计学意义(t=4.001、4.706、4.953,P< 0.05);低钙饮食 氟组肝、肾、脑细胞内[Ca2 ]i水平[(151.57±19.61)、(162.62±31.41)、(158.79±27.65) nmol/L]与低钙饮食对照组[(103.17±14.29)、(103.59±11.40)、(115.26±30.91)nmol/L]比较,差异有统计学意义(t=4.789,P<0.05;t=8.581、6.003,P<0.01);低钙饮食 氟组骨病变明显:细胞内[Ca2 ]i水平与血清离子钙(i[Ca])、总钙(t[Ca])呈负相关(r=-0.59)。结论慢性氟中毒可致机体组织细胞钙超载,低钙可加重氟中毒时的细胞内钙超载,提示细胞内钙超载可能参与氟骨症发病机制并起着重要作用。  相似文献   

6.
目的:探讨重症急性胰膜炎(SAP)肺损伤时血清和肺组织内皮素(ET)变化及丹参的保护作用.方法:Wistar大鼠60只,随机分为假手术组(J 组)、模型组(F组)和丹参治疗组(D组).50 g/L 牛磺胆酸钠胰胆管逆行注射方法制作SAP模型.D组大鼠分别在造模前1 d和造模后10 min ip丹参注射液(5 mL/kg).各组在制模后24 h及 48 h测定血清ET-1水平及其在肺组织(光密度) 表达情况,同时观察肺系数变化及肺组织病理学改变.结果:与J组比较,F组24和48 h肺组织损伤明显加重,血清ET-1水平(F组:75.8±4.8,70.4± 4.8 ng/L;J组:32.0±6.9,30.3±4.8 ng/L)和肺系数显著增高(F组:0.62±0.06,0.73±0.07;J 组:0.41±0.08,0.41±0.07)(P<0.01),肺组织24 和48 h ET-1表达增高(F组:0.48±0.09,0.61± 0.10;J组:0.05±0.01,0.05±0.01)(P<0.01).与 F组比较,D组24和48 h肺组织学损伤明显减轻,血清ET-1水平和肺系数明显下降(60.2± 7.3 ng/L,0.52±0.06,P<0.05;57.9±5.43 ng/L, 0.58±0.06,P<0.01),肺组织ET-1表达明显减少(0.23±0.10,0.36±0.09,P<0.01).相关性分析显示,造模后24和48 h肺组织ET-1表达与肺系数密切相关(r=0.736,P<0.01;r=0.828, P<0.01).结论:ET-1在SAP肺损伤中起着重要的作用, 丹参对SAP肺损伤具有保护作用.  相似文献   

7.
目的探讨IL-1β调控大鼠HSCsTIMP-1mRNA表达与JNK、p38 MAPK通路的关系。方法RT-PCR用于检测大鼠HSCs TIMP-1 mRNA表达;Western blot用于测定IL-1刺激HSCs后JNK、p38的活化程度。结果IL-1β(10ng/mL)作用培养的HSCs 24h后,TIMP-1 mRNA表达(1.191±0.079)明显高于对照组(0.545±0.091),有统计学意义(P〈0.01);IL-1β以时间依赖方式激活JNK、p38通路。用不同浓度JNK阻断剂-SP600125预处理HSCs后,IL-1β上调HSCs TIMP-1 mRNA表达作用受到抑制,分别为10μmol/L,1.022±0.113;20μmol/L,0.8694±0.070;40μmol/L,0.666±0.123,与对照组相比(1.163±0.107),各浓度组均有显著性差异(P〈0.05,P〈0.01,P〈0.01)。用不同浓度p38阻断剂-SB203580预处理HSCs后,HSCs表达TIMP-1 mRNA逐渐增多,分别为10μmol/L,1.507±0.099;20μmol/L,1.698±0.107;40μmol/L,1.857±0.054。与对照组相比(1.027±0.061),各浓度组均有显著性差异(P均〈0.01)。结论IL-1β可通过上调HSCs TIMP-1 mRNA的表达来加速肝纤维化的发生发展,JNK和p38信号蛋白参与了此过程,HSCs中两条通路均可被IL-1激活,但所起作用完全不同,它们相互作用相互调节,共同调控TIMP-1 mRNA的表达。  相似文献   

8.
Fura-2/AM荧光法测定日本血吸虫培养细胞内游离钙离子浓度   总被引:6,自引:0,他引:6  
目的 研究用钙荧光探剂 (Fura- 2 / AM)检测静息状态下日本血吸虫培养细胞内的游离钙离子浓度 ([Ca2 + ]i) ,以及β-巯基乙醇对培养细胞 [Ca2 + ]i的影响。方法 将 18d虫龄的日本血吸虫童虫制成细胞悬液 ,贴壁法接种于 30 ml培养瓶中 ,培养液为 RPMI- 16 4 0含 2 0 %小牛血清及常量抗生素。在培养的 0 - 3d,制备日本血吸虫细胞悬液 ,采用 Fura- 2 / AM钙荧光探剂测定正常静息状态下及加入β-巯基乙醇后日本血吸虫培养细胞内的 [Ca2 + ]i。结果 正常静息状态下 ,培养 0 d的日本血吸虫培养细胞内的 [Ca2 + ]i为 188.2 nmol/ L ;培养 1- 3d的 [Ca2 + ]i两两比较差异无显著性(P>0 .0 5 ) ,它们的平均值为 (187.0± 10 .7) nm ol/ L ,与培养 0 d的 [Ca2 + ]i比较 ,差异也无显著性(P>0 .0 5 )。β-巯基乙醇可使日本血吸虫培养细胞内 [Ca2 + ]i明显升高 (P<0 .0 1) ,并随浓度的增加而升高。结论 培养 1- 3d,静息状态下日本血吸虫培养细胞内的 [Ca2 + ]i比较稳定 ,β-巯基乙醇能影响日本血吸虫培养细胞内的 [Ca2 + ]i  相似文献   

9.
目的 观察钙激活Cl-通道 (ClCa)阻断剂尼氟灭酸 (NFA)对支气管哮喘 (简称哮喘 )气道高反应性的抑制作用。方法 BALB/c小鼠 4 5只 ,按随机数字表法分为哮喘组 (A组 )、吸入NFA预防组 (B组 )和正常对照组 (C组 ) ,每组 15只。检测各组小鼠气道压力峰值 时间指数 (APTI)的差异 ,并比较各组小鼠支气管肺泡灌洗液 (BALF)中内皮素 1(ET 1)和一氧化氮 (NO)的含量。制备A组、C组小鼠的离体完整上皮气管环 (A1、C1组 )和去上皮气管环 (A2 、C2 组 ) ,观察在梯度浓度乙酰甲胆碱(mACh)的激发下 ,5 0 μmol/LNFA预处理对气管环收缩张力的影响。 结果 A组APTI与B组 (mACh为 2 .0mg/kg时 ,两者分别为 1.6 2± 0 .14和 1.2 1± 0 .0 7)比较 ,差异有显著性 (P <0 .0 1) ;A组BALF中ET 1和NO含量分别为 (10 3± 9)ng/L、(48.5± 3.2 ) μmol/L ,B组分别为 (5 3± 5 )ng/L和 (2 3.7± 2 .5 )μmol/L ,A组与B组比较差异有显著性(P <0 .0 1) ;离体气管环收缩张力实验显示 ,C1组完整上皮气管环的张力峰值比为 1.2 6± 0 .14 ,A1组完整上皮气管环的张力峰值比为 3.79± 0 .4 4 ,C2 组去上皮气管环的张力峰值比为 2 .0 6± 0 .18,A2 组去上皮气管环的张力峰值比为 2 .15± 0 .2 1,A1组与C1组、A1组与A2 组比较差异有显著性 (  相似文献   

10.
肝癌细胞中Twist基因表达变化与WNT信号传导通路的关系   总被引:2,自引:1,他引:1  
[摘要]目的探讨肝癌细胞中Twist基因表达与WNT信号传导通路激活的关系。方法将0.05mol/L氯化锂加入人肝癌SMMC7721细胞中培养2h激活VeNT通路,然后分别培养6、12、24h,分别为A1、A2、A3组;将0.1mol/L氯化锂加入人肝癌SMMC7721细胞中培养2h激活WNT通路,然后分别培养6、12、24h,分别为B1、B2、B3组;另设空白对照组。用RT—PCR及荧光定量PCR法检测各组细胞的TwistmRNA。结果lit.PCR法测得TwistmRNA在A1、A2、A3组分别为0.38±0.05、0.60±0.08、0.69±0.06,B1、B2、B3组分别为0.56±O.05、0.59±0.06、0.69±0.04,以上各组细胞TwistmRNA与对照组的0.18±0.03相比,P均〈0.05。荧光定量法测得TwistmRNA在A1、A2、A3组分别为0.43±0.01、0.56±0.01、0.67±0.01,BI、B2、B3组分别为0.52±0.01、0.58±0.00、0.65±0.00,以上各组细胞TwistmRNA与对照组的0.21±0.00相比,P均〈0.05。结论肝癌细胞中Twist基因的表达程度与WNT通路的激活有关。  相似文献   

11.
Relying on a certain degree of abstraction, we can propose that no particular distinction exists between animate or living matter and inanimate matter. While focusing attention on some specifics, the dividing line between the two can be drawn. The most apparent distinction is in the level of structural and functional organization with the dissimilar streams of ‘energy flow’ between the observed entity and the surrounding environment. In essence, living matter is created from inanimate matter which is organized to contain internal intense energy processes and maintain lower intensity energy exchange processes with the environment. Taking internal and external energy processes into account, we contend in this paper that living matter can be referred to as matter of dissipative structure, with this structure assumed to be a common quality of all living creatures and living matter in general. Interruption of internal energy conversion processes and terminating the controlled energy exchange with the environment leads to degeneration of dissipative structure and reduction of the same to inanimate matter, (gas, liquid and/or solid inanimate substances), and ultimately what can be called ‘death.’ This concept of what we call dissipative nature can be extended from living organisms to social groups of animals, to mankind. An analogy based on the organization of matter provides a basis for a functional model of living entities. The models relies on the parallels among the three central structures of any cell (nucleus, cytoplasm and outer membrane) and the human body (central organs, body fluids along with the connective tissues, and external skin integument). This three-part structural organization may be observed almost universally in nature. It can be observed from the atomic structure to the planetary and intergalactic organizations. This similarity is corroborated by the membrane theory applied to living organisms. According to the energy nature of living matter and the proposed functional model, the decreased integrity of a human body's external envelope membrane is a first cause of the structural degradation and aging of the entire organism. The aging process than progresses externally to internally, as in single cell organisms, suggesting that much of the efforts towards the restoration and maintenance of the mechanisms responsible for structural development should be focused accordingly, on the membrane, i.e., the skin. Numerous reports indicate that all parts of the human body, like: bones, blood with blood vessels, muscles, skin, and so on, have some ability for restoration. Therefore, actual revival of not only aging tissue of the human body's membrane, but the entire human body enclosed within, with all internal organs, might be expected. We assess several aging theories within the context of our model and provide suggestions on how to activate the body's own anti-aging mechanisms and increase longevity. This paper presents some analogies and some distinctions that exist between the living dissipative structure matter and inanimate matter, discusses the aging process and proposes certain aging reversal solutions.  相似文献   

12.
Abstract: The effect of swimming at night on rat pineal melatonin synthesis was compared with that of light exposure at night. Rats were forced to swim at 0030 hr (lights out at 2000 hr) and sacrificed by decapitation 15 and 30 min later, immediately after swimming. Other groups of animals were exposed to white light (650μW/cm2) for 15 and 30 min at same time. Swimming caused a rapid and highly significant drop in the melatonin content in the pineal gland; however, the activity of N-acetyltransferase (NAT), the supposed rate limiting enzyme in the melatonin production, was not changed. Despite the drop in pineal melatonin levels, serum concentrations of the indole remained elevated in the rats that swam. In contrast, melatonin levels in the pineal and serum of light exposed rats fell precipitously, accompanied by a significant suppression of NAT activity. Since we anticipated that the strenuous exercise associated with swimming may induce release of artrial natriuretic peptide (ANP) from the heart, which in turn could cause the release of pineal melatonin, in a second study we injected physiological saline intravenously to stretch the cardiac muscle and release ANP. Three milliliters of normal saline was injected during the day into the jugular vein of anesthetized rats that were pretreated with isoproterenol to stimulate pineal melatonin production. Animals were killed 15 min after the saline injection, and pineal NAT activity and pineal melatonin levels were measured. The saline injections caused no alteration in the elevated levels of either NAT or melatonin. These data suggest that the disparity in pineal NAT activity (which was high) and pineal melatonin (which was low), in animals swum at night, may not be caused by ANP which is released during strenuous exercise such as swimming.  相似文献   

13.
The immunoneuroendocrine role of melatonin   总被引:19,自引:0,他引:19  
Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.  相似文献   

14.
Abstract: Well-established circadian physiology supports the view that photoperiodic time measurement utilizes the coincidence between the presence of light and a photosensitive phase of a 'biological clock' to alter reproductive status—the so-called external coincidence model of seasonal breeding. In this review, we examine the mechanism whereby photoperiod interacts with presumed suprachiasmatic nuclei activity to allow endogenous melatonin to normally synchronize reproductive activity to the optimal time of year. The Romney Marsh sheep is particularly explored as an experimental model. It is suggested that the on/off activity of seasonal reproduction may be a robust mechanism able to be predictably manipulated by the judicious use of the light/dark cycle and exogenous melatonin, but firmly based on circadian principles.  相似文献   

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17.
Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.  相似文献   

18.
Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.  相似文献   

19.
Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.  相似文献   

20.
《Indian heart journal》2016,68(4):450-463
The knowledge of variety of chronic total occlusion (CTO) hardware and the ability to use them represents the key to success of any CTO interventions. However, the multiplicity of CTO hardware and their physical character and the terminology used by experts create confusion in the mind of an average interventional cardiologist, particularly a beginner in this field. This knowledge is available but is scattered. We aim to classify and compare the currently used devices based on their properties focusing on how physical character of each device can be utilized in a specific situation, thus clarifying and simplifying the technical discourse.  相似文献   

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