首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 13 毫秒
1.
营养性肥胖对青春期雄性大鼠睾丸生精细胞凋亡的影响   总被引:1,自引:0,他引:1  
目的:探讨营养性肥胖对青春期雄性大鼠睾丸生精细胞凋亡的影响。方法:健康雄性W istar大鼠40只,随机分为两组,每组20只,对照组给予普通饲料喂养,高脂组用高脂、高热量饲料喂养建立营养性肥胖大鼠模型,10周末处死大鼠摘取睾丸。全自动生化分析仪(ACA)检测外周血总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C);光镜观察睾丸组织病理学改变;TUNEL检测睾丸组织凋亡细胞;免疫组化法检测Bc l-2和Bax蛋白的分布和表达;RT-PCR法检测睾丸组织Bc l-2 mRNA和Bax mRNA的表达。结果:高脂组TC、TG、HDL-C和LDL-C(5.17±0.17、1.18±0.09、1.76±0.11、5.08±0.18)较对照组(1.38±0.12、0.39±0.05、0.97±0.07、0.75±0.06)显著升高(P<0.05);高脂组生精细胞凋亡指数(37.17±2.74)较对照组(5.16±0.81)显著升高(P<0.01),凋亡细胞以精原细胞和精母细胞为主;高脂组Bax蛋白和Bax mRNA(153.26±8.74、1.08±0.12)表达较对照组(101.81±6.14、0.37±0.04)明显升高(P<0.01);高脂组Bc l-2蛋白和Bc l-2 mRNA(139.26±7.21、0.46±0.05)表达较对照组(159.37±8.96、1.05±0.11)明显降低(P<0.01)。结论:营养性肥胖诱导大鼠睾丸生精细胞凋亡增多,其机制可能与Bc l-2表达降低和Bax表达升高相关。  相似文献   

2.
The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and its receptors in varicocele-induced testicular dysfunction in an experimental rat model. The rats were divided into three groups: control, sham and varicocele. Varicoceles in rats were induced by partial ligation of the left renal vein and left testes. The rats were analyzed 13 weeks after surgery. The degree of DNA fragmentation within cells in the testis was determined using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) assay. Tubule degeneration was evaluated using the Johnsen score. The expression of TRAIL and its receptors was detected by immunohistochemical and Western blotting techniques. The apoptotic index, Johnsen score and the expression of TRAIL and TRAIL receptors were examined. The data are presented as the mean±s.d. and were analyzed using computer software. The Kruskal–Wallis and Dunn''s multiple comparison tests were used in the statistical analyses. The germ cell apoptotic index was increased in rats with varicoceles when compared with the sham and control groups (P=0.0031). The Johnsen score was significantly decreased in the varicocele group when compared with the sham and control groups (P<0.0001). Immunohistochemical and Western blotting analyses showed that after varicocele induction, the expression of TRAIL-R1 and TRAIL-R4 in germ cells was increased and the expression of TRAIL-R2 was decreased. There are no significant differences among the groups in terms of TRAIL and TRAIL-R3 receptor expression. The results of this study indicate that TRAIL and its receptors may have a potential role in the pathogenesis of varicocele-induced testicular dysfunction.  相似文献   

3.
4.
Objective:   To investigate the effects of strontium fructose 1,6-diphosphate (FDP-Sr) on testicular dysfunction induced by diabetes.
Methods:   Diabetes was induced by a single injection of streptozotocin (65 mg/kg, i.p.). After 28 days, therapy with three doses (50, 100, and 200 mg/kg per day, p.o.) of FDP-Sr was carried out for another 4 weeks.
Results:   The rats exhibited morphological lesions of testes and significant decreases in serum testosterone levels after 2 months of diabetes. Testicular tissues of diabetic rats showed significantly increased malondialdehyde levels and declined glutathione peroxidase activity. Meanwhile, augmented DNA fragmentation was observed, along with downregulated Bcl-2 and upregulated Bax expressions at both mRNA and protein levels. FDP-Sr showed significant antioxidant effects in both in vitro and in vivo experiments, and significantly relieved apoptosis and the decline of serum testosterone caused by diabetes.
Conclusions:   Testicular injury and apoptosis induced by diabetes are partially attributed to the augmented oxidative stress in testicular tissue. FDP-Sr indirectly alleviates these pathologic alterations by suppressing the generation of reactive oxygen species.  相似文献   

5.
单侧隐睾大鼠对侧睾丸的损害与Bcl-2和Bax基因表达   总被引:2,自引:2,他引:2  
目的:探讨单侧隐睾大鼠对侧睾丸生精细胞凋亡与Bcl-2/Bax基因表达的关系。方法:20只健康SD雄性大鼠(22日龄)随机分成隐睾组和对照组,每组10只。通过手术建立单侧隐睾动物模型。术后90 d取对侧睾丸,采用原位缺口末端标记(TUNEL)法检测生精细胞凋亡,免疫组化SP法检测Bcl-2/Bax基因表达。结果:与对照组相比,隐睾组对侧睾丸生精细胞凋亡显著增多(P<0.01),重量显著减轻(P<0.01),Bax表达显著升高(P<0.01),Bcl-2表达显著降低(P<0.01)。凋亡细胞主要是初级精母细胞和圆形精子细胞。结论:单侧隐睾大鼠对侧睾丸的生精细胞凋亡增多与Bcl-2基因表达降低、Bax基因表达升高密切相关。细胞内Bc l-2/Bax比值是影响生精细胞凋亡的重要因素之一。  相似文献   

6.
In this study, it was aimed to investigate possible ameliorating effects of thymoquinone on testicular damage in an epilepsy model. Adult male Wistar rats were divided into 4 groups. The animals in sham-operated groups were given saline or thymoquinone (s.c.); and the animals in pentylenetetrazole (PTZ) group were applied PTZ. The animals in PTZ+thymoquinone group were given thymoquinone (i.p) for 6 days after applying PTZ. Hematoxylin-eosin, periodic acid–Schiff and TUNEL staining and PCNA, StAR, inhibin β-B immunohistochemistry and ZO-1 immunofluorescence methods were applied. Staining intensity and cell numbers were determined. Degeneration of seminiferous tubules was observed in PTZ group. Most of the tubules showed normal morphology in the PTZ+thymoquinone group. Apoptotic cell index was found to be increased and proliferative index decreased in PTZ group. Thymoquinone administration decreased apoptotic index and increased proliferation index. In PTZ group, ZO-1, StAR and inhibin β-B immunohistochemical staining intensity was observed to be decreased and after thymoquinone application, ZO-1 was increased. StAR and inhibin β-B-positive cell numbers were decreased in PTZ group and increased in the PTZ +thymoquinone group. In this study, it was observed that PTZ-induced epileptic seizures caused testicular damage in the rat and thymoquinone ameliorated these effects.  相似文献   

7.
The present study aimed to explore the impact of onion (Allium cepa Linnaeus) extract on testicular damage induced by dexamethasone. Forty male Wistar rats were divided into four groups (control, dexamethasone, onion extract and dexamethasone group treated with onion extract). Testosterone levels, antioxidant parameters and the expression of caspase-3 and IL-1β, IL-12, IL-10 genes, as well as histopathological examination and immunohistochemical studies of Bcl2 and caspase-9 expression, were examined. Dexamethasone was found to decrease GSH, total antioxidant activity and testosterone levels, meanwhile treatment with onion extract normalised these levels. MDA was increased in dexamethasone group but appeared normal in the treated group. Dexamethasone was shown to downregulate IL-10 and IL-2 gene expression. Conversely, IL-1β and caspase-3 gene expression were upregulated by dexamethasone and normalised in the treated group. Histopathological analysis found that dexamethasone caused atrophy to the seminiferous tubules and degeneration to spermatocytes, and immunohistochemical analysis showed overexpression of caspase-9 and inhibited the expression of Bcl-2 in dexamethasone group. These effects were normalised in the onion extract treated group. In conclusion, onion extract have a preventative effect against dexamethasone-induced testicular damage in rats; therefore, its use in complementary therapy is recommended.  相似文献   

8.
This study was conducted to investigate the effects of aescin treatment in a rodent model treated with an experimentally induced varicocele. Experimental varicocele was induced by partial ligation of the left renal vein of rats. Aescin administration was performed daily for 4 weeks after the varicocele induction. Seven weeks later, a contrast‐enhanced ultrasound was performed of the rats' testis to assess testicular blood flow. The animals were sacrificed, and H&E staining was then used to evaluate testicular pathological changes and polymorphonuclear leucocytes density. Cauda epididymal sperm counts and motility were evaluated. Blood was collected for the measurement of follicle‐stimulating hormone, luteinising hormone and testosterone. Contrast‐enhanced ultrasound showed that there were significant decreases in testicular blood flow in the aescin‐treated groups compared with those in control varicocele group. Testicular oedema was detected in those rats treated with a varicocele but without aescin, while no oedema was found in the experimental group. H&E staining showed dysfunctional spermatogenesis in both cohorts; however, polymorphonuclear leucocytes density was significantly reduced in aescin‐treated groups. There was an increase in sperm counts of the aescin‐treated groups. Our study demonstrated that aescin could exert therapeutical effects on reversal of testicular lesions in varicocele rats.  相似文献   

9.
目的 探讨鼻咽腔降温对大鼠全脑缺血-再灌注损伤的保护作用.方法 健康雄性Wistar大鼠18只,随机均分为三组:全身降温组(A组)、鼻咽腔降温组(B组)、常温组(C组).采用Pulsinelli四血管阻断法建立大鼠伞脑缺血-再灌注损伤模璎.A、B组海马温度降至靶温度33℃.阻断双侧颈总动脉20 min再灌注,低温维持1 h后复温.C组保持大鼠直肠和海马温度均为(37±0.5)℃,阻断双侧颈总动脉20 min再灌注.利用微透析技术收集大鼠海马CA1区微透析液并测其谷氨酸(Glu)含量.再灌注8 h后断头取脑,用免疫组化法观察海马CA1区Bcl-2和Bax的表达.结果 A组海马温度从37℃降到33℃所需时间为B组的4.8倍;缺血10、20 min、再灌注后10、20、30 min时A、B组Glu含量明显低于C组(P<0.05);A、B、C组海马CA1区Glu含量恢复至缺血前水平所需时间分别为(19.92±1.30)、(20.17±1.34)、(39.67±1.49)min.与C组比较,A、B组的Bax免疫阳性细胞数显著减少(P±0.05);A、B组的Bcl-2免疫阳性细胞数显著增多(P<0.05).结论 鼻咽腔降温对大鼠全脑缺血-再灌注损伤具有保护作用,且鼻咽腔降温法降低海马温度的速度明显快于全身降温法.  相似文献   

10.
PURPOSE: We investigated the effect of varicocele repair on testicular volume according to age in children and adolescents and review the long-term results of varicocele surgery. MATERIALS AND METHODS: The study included 39 boys 11 to 19 years old with clinical palpable varicocele who underwent varicocele surgery with at least 1 year of postoperative followup. Preoperative and postoperative testicular volumes were monitored and measured with an ellipsoid Prader orchidometer. Physical examination findings (testicular volumes and testicular consistency) in all boys, and serum hormone values and semen parameters in 16 adolescents were recorded and compared before and after surgery. RESULTS: Left unilateral varicocelectomy was done in 29 boys (74%) and bilateral varicocelectomy in 10 (26%). While no postoperative hematoma, infection or testicular atrophy was observed, 1 boy (2.5%) had varicocele recurrence and 2 boys (5.1%) had minimal hydroceles that required no intervention. Significant increases were observed in postoperative sperm concentration (p = 0.01), total motile sperm count (p = 0.009), testis volume (p = 0.000) and serum testosterone level (p = 0.014). All 15 boys with preoperative soft testis had normal testicular consistency postoperatively. Of the 19 boys with preoperative testicular atrophy 10 (53%) did regain normal testicular growth, while 9 (47%) retained testicular volume loss after surgery. When comparing preoperative to postoperative increase in testicular volume according to age in all boys, the mean was statistically significantly higher in boys younger than 14 years (left testis p = 0.037, right testis p = 0.000). CONCLUSIONS: Testicular consistency achieved normal firmness after varicocelectomy in all boys with preoperative soft testis. While there was catch-up growth in comparison to the contralateral testis, testicular consistency improved but testicular volumes may not increase significantly after varicocele repair at ages older than 14 years. However, in these adolescents postoperative semen parameters and serum hormone values may significantly improve regardless of testicular volume. Therefore, boys with varicocele and their families should be fully informed in light of these findings.  相似文献   

11.
目的 观察糖尿病(diabetes mellitus,DM)大鼠胰岛素治疗后阴茎勃起功能的情况及阴茎海绵体细胞凋亡、Bcl-2和Bax的基因表达.方法 成年雄性Wistar大鼠50只.腹腔内注射链尿佐菌素(STZ)建立糖尿病动物模型,成模后8周根据阴茎勃起功能的情况,随机分成未行胰岛素治疗组(DM组)和胰岛素治疗组.胰岛素治疗组大鼠治疗16周后,处死并取阴茎海绵体,测血糖及糖化血红蛋白,用原位末端标记法(TUNEL)检测细胞凋亡,western blot方法检测阴茎组织Bcl-2、Bax蛋白的表达.结果 DM组大鼠阴茎勃起功能明显低于胰岛素组.胰岛素组血糖及糖化血红蛋白下降(P<0.001),DM组大鼠较胰岛素治疗组大鼠阴茎海绵体凋亡细胞数明显增多,Bax表达增强,Bcl-2表达减弱.结论 DM大鼠胰岛素治疗后,阴茎海绵体细胞凋亡率减少,阴茎勃起功能改善,Bcl-2和Bax可能参与了DM大鼠阴茎海绵体细胞凋亡的基因调控.  相似文献   

12.
We evaluated the development of embryos generated from the fertilisation of oocytes with spermatozoa isolated from animals with primary testicular damage (PTD). Embryos derived in vivo or in vitro from oocytes fertilised with spermatozoa produced by PTD rats that had undergone surgical treatment for the PTD (group A1), or PTD rats (group A2), or control rats (group B) were cultured and transferred to recipients. At the end of the experimental period, the fertilisation potential of each rat was assessed in vitro (IVF trials). Sperm 8-oxodG/dG ratio (a marker of DNA oxidative status) was significantly larger in group A2 than in groups A1 and B. Blastocysts of the group A2 transferred to recipients demonstrated a significantly larger loss before implantation than transferred blastocysts of groups A1 or B. In addition, the proportion of implanted blastocysts that could not complete the intrauterine development was significantly larger in group A2 than in groups A1 and B. This study reveals a post-fertilisation detrimental effect in animals with PTD on the capacity of oocytes (fertilised either in vitro or in vivo ) to develop in vitro and implant after transferring them to recipients probably attributable to sperm DNA oxidative damage.  相似文献   

13.
This study was conducted to survey the protective effect of pre‐treatment with Persian honey during post‐ischaemia reperfusion on ischaemia–reperfusion (IR)‐induced testis injury. Animals were divided into four groups of IR, honey + ischaemia‐ reperfusion (HIR), vitamin C + ischaemia‐ reperfusion (VIR) and carbohydrates + ischaemia‐ reperfusion (CIR). The testes were examined for spermatogenesis index. Detection of single‐ and double‐stranded DNA breaks at the early stages of apoptosis was performed. Total serum concentration of FSH, LH and testosterone was measured using ELISA. All data were expressed as mean ± SD in each group, and significance was set at p ≤ .05. Spermatogenesis index was significant in the HIR group (p < .001). Serum levels of FSH and LH were significantly higher in the CIR and HIR groups. Serum levels of testosterone were significantly higher in VIR and HIR groups. Apoptotic cells in IR and CIR groups increased significantly statistically (p < .001), while in HIR and VIR groups, the number of apoptotic cells decreased and the positive cells of TUNEL staining were detected in spermatocytes and spermatid. The present study indicates that honey decreases the cellular damage and apoptosis during testicular I/R injury, with significant protective effects on reproductive hormone production.  相似文献   

14.
With increasing evidence for faulty paternal contribution to reproduction, there has been a steady increase in studies highlighting an association between sperm DNA damage, failed/delayed fertilisation and aberrant embryo development. Owing to prevailing ambiguity, the aims of the study were to analyse the genetic integrity of the male gamete and then to understand its association with age, standard semen parameters, lifestyle and occupational factors. The study included 504 subjects, attending university infertility clinic for fertility evaluation and treatment. Semen characteristics were analysed by standard criteria; terminal deoxynucelotidyl transferase-mediated nick end-labelling assay was employed for DNA damage assessment. The average incidence of sperm DNA damage in patients with normozoospermic semen parameters was <10%. Patients with oligozoospermia, severe oligozoospermia, oligoasthenoteratospermia, asthenoteratozoospermia and necrozoospermia had significantly higher level of sperm DNA damage (P < 0.001). Patients above 40 years of age had significantly high levels of DNA damage (P < 0.001) compared with their counterparts. Patients with varicocele and a history of alcohol consumption had higher incidence of spermatozoa with DNA damage (P < 0.01). Poor sperm characteristics in the ejaculate are associated with increased sperm DNA damage. Age-related increase in sperm DNA damage and association of the same with varicocele and alcohol consumption are also demonstrated.  相似文献   

15.
Varicocele is a rather common andrological condition in adolescents, which can adversely affect testicular growth and seminal parameters, leading to infertility in about 20% of adults. The aim of this study was to investigate if treating varicocele before the age of 18 is a beneficial option to improve testicular hypotrophy and seminal parameters and if minimally invasive techniques could be an appropriate treatment choice for adolescent varicocele associated with spermatic vein reflux. A percutaneous scleroembolization was performed in sixty‐four consecutive young patients (13–19 years old) with left varicocele, preceded by a fluoroscopy. In thirty‐four of them, semen samples were also collected. We examined semen samples and testicles dimensions before and after percutaneous varicocelectomy, compared to a nonoperated control group, with a six‐month follow‐up. Total sperm count and sperm morphology were significantly increased in the intervention group. Left testicular volume significantly increased in both groups, while only correction of varicocele improved spermatozoa release per unit of testis volume. We conclude that early varicocelectomy by percutaneous scleroembolization significantly ameliorates seminiferous tubules activity in the critical adolescent phase of testicular growth. It is suggested that adolescents should be; offered varicocele repair as soon as possible in order to improve reproductive potential.  相似文献   

16.
Lead (Pb) is a nonthreshold multi‐targeted toxicant that causes alterations in different organs of the body, especially the gonads. This study was aimed to investigate the possible protective effect of thymoquinone (TQ), the major active ingredient of volatile oil of Nigella sativa seeds, against Pb‐induced testicular histological damage. Adult male rats were randomised into four groups as follows: control group received no treatment, Pb group was exposed to 2000 ppm of Pb acetate in drinking water, Pb‐TQ group was cotreated with Pb plus TQ (5 mg/kg/day, per os) and TQ group receiving only TQ. All treatments were applied for five weeks. Results showed that Pb exposure produced morphological changes in the testis, especially degeneration of germinal epithelium, sloughing of germ cells into the lumen of seminiferous tubules and reduction in the number of luminal spermatozoa. Interestingly, coadministration of TQ to the metal‐treated animals prevented the testicular adverse effects. In conclusion, our data indicate for the first time a remarkable protective effect of TQ against Pb‐induced testicular histopathological lesions in rat. On this basis, TQ deserves more consideration and further examination as a potential therapeutic option.  相似文献   

17.
目的 评价不育男性双侧I度精索静脉曲张对睾丸体积和生殖激素水平的影响.方法 185例不育男性双侧I度精索静脉曲张(A组)和149例正常生育男性(B组),比较其睾丸体积、卵泡刺激素(FSH)、黄体生成素(LH)和睾酮(T)水平.结果 A组患者两侧睾丸体积均小于B组,但睾丸体积绝对差异和睾丸体积相对差异与B组比较,无统计学意义.A组患者血清FSH水平高于B组,而LH、T与B组相比,差异无统计学意义.结论 不育男性双侧I度精索静脉曲张可导致患者双侧睾丸体积减小,血清FSH水平升高.  相似文献   

18.
We evaluated the efficiency of microdissection testicular sperm extraction (MicroTESE) in patients with nonobstructive azoospermia (NOA) and their pregnancy outcomes in a programme based on in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI). Fifty-six MicroTESE procedures were performed in 53 patients with NOA. Pre-operative levels of luteinising hormone, follicle-stimulating hormone (FSH), testosterone and prolactin were obtained and a Doppler sonography examination was conducted. Sperm retrieval rate, mean age of female partner, mean ICSI and fertilisation rate, number and quality of embryos transferred, implantation, pregnancy and miscarriage rates were calculated. Samples for testicular histological analysis were taken trans-operatively in every case. Sperm retrieval rate, mean ICSI per case and fertilisation rate were 57.1%, 7.4% and 58.4% respectively. A significant difference in pre-operative testicular volume ( P  = 0.001), serum FSH ( P  = 0.008) and total testosterone levels ( P  = 0.021) was found in patients from whom sperm could be retrieved. Mean 1.9 type A embryos were transferred per cycle. Implantation, clinical pregnancy and miscarriage rates were 20%, 40% and 18.7% respectively. It is concluded that MicroTESE is a viable option for men with NOA, offering excellent results in couples undergoing IVF-ICSI. Pre-operative serum FSH, testicular volume and total testosterone levels may have a prognostic value, although more data are needed to determine their significance and whether or not patients should be excluded from an initial sperm retrieval attempt.  相似文献   

19.
The aim of this study was to investigate the effects of Achillea millefolium extract in paclitaxel-induced testicular toxicity in rats. The groups were designed as (1) control, (2) paclitaxel (8 mg/kg, intraperitoneally), (3) paclitaxel (8 mg/kg, intraperitoneally) + Achillea millefolium (200 mg/kg, orally for 14 consecutive days) and (4) paclitaxel (8 mg/kg, intraperitoneally) + Achillea millefolium (400 mg/kg, orally for 14 consecutive days). Serum levels of testosterone, luteinising hormone and follicle-stimulating hormone, as well as total antioxidant capacity and total oxidant status were measured one day after receiving the last dose of Achillea millefolium extract. Testicular superoxide dismutase activity, malondialdehyde, tumour necrosis factor alpha and interleukin-1β levels, the expressions of nuclear factor kappa B and caspase-3 were evaluated. In addition, testicular sections were evaluated histopathologically and 8-hydroxy-2′-deoxyguanosine was detected immunohistochemically. Achillea millefolium improved the levels of luteinising hormone, follicle-stimulating hormone and testosterone, upregulated testicular antioxidant enzymes and downregulated inflammation. Furthermore, we observed that Achillea millefolium restored testicular histopathological structure and significantly suppressed oxidative DNA damage and apoptosis by reducing the expression of caspase-3. Taken together, our results suggest that Achillea millefolium has protective effects against paclitaxel-induced testicular toxicity and is a promising natural product with the potential to improve male fertility.  相似文献   

20.
糖尿病大鼠阴茎海绵体细胞凋亡与Bcl-2和Bax基因表达   总被引:1,自引:0,他引:1  
目的 :观察糖尿病 (DM)大鼠阴茎海绵体细胞凋亡、Bcl 2和Bax的基因表达及两者的相关性。 方法 :成年雄性Wistar大鼠 5 0只 ,随机分为正常对照组 (NDM组 ) 10只和造模组 4 0只。造模组以 5 0mg/kg尾静脉注射 2 %四氧嘧啶 (AXN) ,未发生DM者 ,为AXN组 ;成为DM模型者为DM组。大鼠饲养 8周后处死并取阴茎海绵体 ,用原位末端标记法 (TUNEL)检测细胞凋亡 ,用免疫组化方法检测Bcl 2、Bax的基因表达。 结果 :DM组大鼠较NDM组大鼠阴茎海绵体凋亡细胞数明显增多 (P <0 .0 1) ,Bax表达亦增强 (P <0 .0 1) ,Bcl 2表达减弱 (P <0 .0 1) ,Bcl 2 /Bax比值降低。 结论 :DM大鼠阴茎海绵体细胞凋亡率增加 ,细胞凋亡可能是糖尿病性勃起功能障碍的发病机制之一 ,Bcl 2和Bax可能参与了DM大鼠阴茎海绵体细胞凋亡的基因调控。  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号