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1.
刘海英  陈峰  姚嘉 《实用医学杂志》2023,(22):2878-2884
目的 研究抑制miR-767-5p对乳腺癌细胞增殖、迁移、侵袭和上皮间充质转化(EMT)的影响。方法 体外培养乳腺癌细胞,将miR-767-5p inhibitor和inhibitor-NC转染到乳腺癌细胞中。细胞分为3组:空白对照组(Control组),miR-767-5p inhibitor组和inhibitor-NC组,采用CCK-8法、Transwell法、划痕实验检测miR-767-5p抑制剂对乳腺癌细胞增殖、侵袭和迁移能力的影响。Western blot检测迁移相关蛋白基质金属蛋白酶-2(MMP-2)和MMP-9的表达水平。双荧光素酶报告基因实验验证miR-767-5p和胰岛素样生长因子-1(IGF1)的靶向结合作用。RT-qPCR和Western blot实验检测IGF1 mRNA和蛋白的表达水平。采用Western blot检测EMT相关蛋白的表达水平。结果 CCK-8实验结果显示,与Control组和inhibitorNC组相比,miR-767-5p inhibitor组显著抑制MCF-7细胞的增殖(P <0.05)。Transwell实验结果表明,与Contr...  相似文献   

2.
目的了解p53基因变异和B细胞淋巴瘤甘氨酸脱羧酶(GLDC)表达的关系。 方法基于免疫组化染色结果,BALB/c裸鼠分为p53蛋白阳性组和p53蛋白阴性组。采用实时荧光定量PCR检测B细胞淋巴瘤组织甘氨酸脱羧酶mRNA表达,采用免疫印迹实验检测B细胞淋巴瘤组织甘氨酸脱羧酶蛋白表达。设计GLDC特异性siRNA转染人B淋巴瘤细胞系Raji细胞。采用t检验比较GLDC mRNA和GLDC蛋白表达组间的差异,并采用方差分析比较阴性对照siRNA组、空白对照组和GLDC siRNA转染组增殖能力的差异。 结果相对于p53蛋白阴性组,p53蛋白阳性组GLDC mRNA和蛋白表达都升高,GLDC mRNA表达在p53蛋白阴性组为2.38±0.26,在p53蛋白阳性组升高到11.32±1.65,差异具有统计学意义(t=6.74,P<0.01)。相对于阴性对照siRNA组(0.61±0.06)和空白对照组(0.51±0.04),GLDC siRNA转染组(0.22±0.01)细胞增殖能力明显降低,差异具有统计学意义(t=3.74,P<0.05), 结论p53基因变异和B细胞淋巴瘤GLDC表达有直接的关系。GLDC基因能促进p53基因变异B细胞淋巴瘤增殖。GLDC的研究可能为治疗B细胞淋巴瘤提供新的靶点。  相似文献   

3.
目的探讨结直肠癌Fas相关死亡域样白介素-1β转换酶抑制蛋白(FLIP)(S)mRNA的表达水平及其临床意义。方法建立检测FLIP(S)mRNA荧光定量逆转录聚合酶链反应(FQ-RT-PCR),用该方法检测结直肠癌组织及其癌旁正常组织FLIP(S)mRNA表达水平。结果 FLIP(S)mRNA在结直肠癌组织和癌旁正常组织都有表达,在结直肠癌组织表达水平为0.23±0.19,明显高于癌旁正常组织0.13±0.10(P<0.05)。FLIP(S)mRNA表达水平与结直肠癌组织学分级密切相关(P<0.05),肿瘤分化程度越低,FLIP(S)mRNA的表达水平越高。结论 FLIP(S)在结直肠癌组织中表达升高;在分化越低组织中其表达水平越高;为揭示结直肠肿瘤的发生、发展提供新的理论依据,并为结直肠癌的有效治疗提供新的思路。  相似文献   

4.
目的 探讨Notch3信号通路对三阴性乳腺癌细胞株BT20活性的影响及分子机制。方法 采用小干扰RNA(siRNA)转染技术,下调BT20细胞中Notch3 mRNA表达,MTT法检测转染后5 d的细胞增殖情况,并进行Transwell细胞迁移和侵袭实验;RT-PCR检测BT20细胞中CyclinD1、BCL-2、BCL-xl mRNA表达水平;Western blot检测BT20细胞核内NF-κB蛋白表达水平。结果 培养第2天、第3天、第4天、第5天时,Notch3 siRNA组细胞增殖水平明显低于空白对照组与阴性对照组,差异均有统计学意义(t分别=3.15、4.03、2.94、3.26、4.01、3.50、2.97、3.11,P均<0.05);随着培养时间的延伸,三组细胞增殖水平均逐渐增加,差异均有统计学意义(F分别=5.94、7.65、9.76,P均<0.05)。Notch3 siRNA组迁移细胞数、侵袭细胞数均明显低于空白对照组和阴性对照组,差异均有统计学意义(t分别=5.32、6.02、4.97、5.07,P均<0.05);空白对照组与阴性对照组迁移细胞数、...  相似文献   

5.
摘要:目的:观察中性粒细胞明胶酶相关脂质运载蛋白(NGAL)对缺氧/复氧肾小管上皮细胞HK-2自噬水平的影响,探讨NGAL在肾小管上皮细胞缺氧/复氧过程中发挥的作用。 方法:构建高表达NGAL的载体pcDNA3.1(-)/myc-His B-NGAL,并转染HK-2细胞。将HK-2细胞分为未转染组和转染组,每组再分为缺氧/复氧处理组和未处理组。用western blot法检测转染后NGAL蛋白表达水平和缺氧/复氧后细胞自噬微管相关蛋白轻链3-Ⅱ(LC-3-Ⅱ)表达水平,并用CellTiter Blue细胞活性检测系统检测细胞活力。 结果:成功构建高表达NGAL的载体pcDNA3.1(-)/myc-His B-NGAL,转染HK-2细胞后,细胞NGAL蛋白表达水平高于未转染组(t=8.959,P<0.05)。缺氧1 h/复氧24 h后,转染组细胞LC-3-Ⅱ表达水平高于未转染组(t=21.721,P<0.05),而细胞活力低于未转染组(t=-6.238,P<0.05)。 结论:在HK-2细胞缺氧/复氧状态下,NGAL能增强细胞自噬水平,导致细胞损伤加重甚至死亡。  相似文献   

6.
本研究旨在检测不同亚型急性淋巴细胞性白血病患者血清PTK7 mRNA和蛋白的表达水平,探讨其诊断的临床价值.选择2012年5月-2014年4月在本院诊断为急性淋巴细胞白血病(ALL)患者,按照纳入和排除标准共纳入136例患者,按照细胞形态学将他们分成L1组(n=42)、L2组(n=45)和L3组(n=49),另外选择48例正常儿童为对照组.使用荧光定量PCR检测外周血单个核细胞中PTK7 mRNA转录水平,采用Westem blot检测血清中PTK7蛋白表达量.结果表明,L1组PTK7转录水平显著高于正常组(P=0.000).L2组PTK7转录水平显著高于L1组(P =0.000).L3组与L2组之间无显著性差异(P =0.123).L1组患者PTK7蛋白水平显著高于正常组(仁1.564,P=0.000),L2组PTK7蛋白水平显著高于L1组(t=1.437,P=0.003),L3组PTK7蛋白水平与L2组比较无显著差异(t=0.821,P=0.312).结论:血清PTK7蛋白表达检测具有诊断ALL的潜在临床价值,但对ALL分型不具有型特异性.  相似文献   

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目的:检测细胞凋亡、Fas/FasL mRNA及其蛋白在特发性肺纤维化患者肺泡/支气管上皮细胞的表达,探讨细胞凋亡和促凋亡信号在此疾病中的意义。方法:应用末端原位杂交(TUNEL)、原位杂交、免疫组化技术,对12例特发性肺纤维化患者的肺活检组织(IPF组)及10例正常肺组织(对照组)检测了细胞凋亡、Fas/FasL mRNA及其蛋白表达的变化。结果:特发性肺纤维化组肺泡/支气管上皮细胞凋亡指数上调,明显高于对照组(P〈0.01);IPF组肺泡/支气管上皮细胞中Fas/FasL mRNA及其蛋白表达上调,高于对照组(P〈0.01);IPF组肺泡/支气管上皮细胞凋亡指数与其Fas/FasL mRNA及蛋白表达之间均无明显相关(P〉0.05)。结论:肺纤维化时肺泡/支气管上皮细胞凋亡上调,Fas/FasL mRNA及其蛋白表达增强,在肺纤维化发生、发展中起重要作用。  相似文献   

8.
吕艳  郑旭  韩燕燕  高珊  李翀  耿强 《实用医学杂志》2023,(22):2898-2902
目的 探讨CCN5基因敲降对人乳腺癌细胞增殖的影响;探索CCN5在人乳腺癌细胞增殖中的作用机制;探寻CCN5与乳腺癌发生、发展中的相关性。方法 设置CCN5siRNA转染组,慢病毒空载体转染组,空白对照组,分别采用不同处理方法。CCN5siRNA转染组(以下简称“转染组”):慢病毒载体介导的CCN5siRNA转染人乳腺癌细胞MCF-7;慢病毒空载体转染组(以下简称“空载组”):慢病毒空载体转染人乳腺癌细胞MCF-7;空白对照组不进行任何转染操作。采用MTT法检测各组细胞的增殖能力变化,采用RT-PCR、Western blot检测各组细胞CCN5、Skp2和P27Kip1mRNA及蛋白表达。结果 MTT实验表明:转染组细胞增殖力高于空载组和空白对照组;RT-PCR和Western blot结果表明:转染组CCN5mRNA和蛋白水平低于空载组和空白对照组,差异有统计学意义(P <0.05);转染组Skp2、P27Kip1mRNA和蛋白水平高于空载组和空白对照组,差异有统计学意义(P <0.05)。结论 CCN5siRNA抑制CCN5mRNA和蛋白表达,促进乳腺癌细胞增殖,并且...  相似文献   

9.
目的探讨miR-9表达对SGC-7901胃癌细胞株侵袭力的影响及其可能的作用机制。 方法培养SGC-7901胃癌细胞株,分组转染miR-9阴性对照(miR-9 NC)、miR-9模拟物(miR-9 mimics)和miR-9抑制剂(miR-9 inhibitor),上调或下调细胞中miR-9表达水平。采用Transwell实验评价胃癌细胞侵袭能力,应用SYBR Green荧光实时定量聚合酶链反应和Western blot实验检测SGC-7901细胞中MMP-2 mRNA和蛋白表达水平。对miR-9空白对照组、miR-9 mimics组和miR-9 inhibtor组miR-9的相对表达量、MMP-2 mRNA及蛋白相对表达量,Transwell侵袭实验SGC-7901细胞中侵袭细胞数量,多组间均数比较采用单因素方差分析。 结果与空白对照组SGC-7901细胞miR-9的相对表达量比较:转染miR-9 mimics组SGC-7901细胞miR-9的相对表达量明显上升,差异具有统计学意义[(1.002±0.083)vs(15.375±3.097),P=0.012];转染miR-9 inhibitor组SGC-7901细胞miR-9的相对表达量明显下降,差异具有统计学意义[(1.002±0.083)vs(0.279±0.039),P=0.022]。与空白对照组SGC-7901细胞中侵袭细胞数量比较:转染miR-9 mimics组SGC-7901细胞中侵袭细胞数量明显减少,差异具有统计学意义[(72.0±2.2)vs(23.6±4.2),P=0.026];转染miR-9 inhibitor组SGC-7901细胞中侵袭细胞数量明显增加,差异具有统计学意义[(72.0±2.2)vs(102.4±7.6),P=0.037]。与空白对照组SGC-7901细胞中MMP-2的mRNA及蛋白的相对表达量比较:miR-9 mimics组SGC-7901细胞中MMP-2的mRNA及蛋白相对表达量明显下降,差异具有统计学意义[(1.006±0.133)vs(0.371±0.132),P=0.011;(0.573±0.063)vs(0.261±0.159),P=0.024];miR-9 inhibitor组SGC-7901细胞中MMP-2的mRNA及蛋白相对表达量明显上升,差异具有统计学意义[(1.006±0.133)vs(2.995±0.701),P=0.015;(0.573±0.063)vs (0.708±0.079),P=0.016]。 结论在SGC-7901胃癌细胞株中,miR-9可下调MMP-2的表达,抑制细胞的侵袭能力。  相似文献   

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摘要:目的:检测Slug基因在宫颈癌和慢性宫颈炎症组织中的表达,探讨其与宫颈癌发生、发展的相关性及在宫颈癌诊疗中的意义。 方法:收集31例宫颈癌组织和17例慢性宫颈炎症组织标本,用RT-PCR、实时定量PCR检测Slug mRNA的表达;用western blot分析Slug蛋白的表达水平,并分析Slug mRNA在宫颈癌中的表达水平与临床病理因素及高危型HPV病毒载量的相关性。 结果:宫颈癌组织中SlugmRNA表达阳性率及相对表达水平均显著高于慢性宫颈炎症组织(χ2=9.872;U=140,P均<0.01);western blot也证实Slug蛋白表达水平高于慢性宫颈炎症组织(U=98,P<0.01)。在宫颈癌组织中,Slug的阳性表达与国际妇产科学联盟(International Federation of Gynecology and Obstetrics, FIGO)肿瘤分期相关,Ⅱ期阳性率高于Ⅰ期(χ2=4.037,P<0.05),与年龄、肿瘤大小、分化程度、淋巴细胞转移和病理类型无关;而Slug基因相对表达水平与高危型HPV病毒载量呈正相关(r=0.554,P<0.01)。 结论:宫颈癌组织中Slug的表达上调,且与宫颈癌的FIGO分期相关,在宫颈癌的发生机制中,可能存在Slug和高危型HPV的协同作用。  相似文献   

11.
This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.  相似文献   

12.
目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高<20%视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P<0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P<0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P<0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.  相似文献   

13.
The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.  相似文献   

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Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.  相似文献   

16.
17.
Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.  相似文献   

18.
目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4%(55/72)]明显高于成功组[31.7%(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3%(42/72)]高于成功组[38.1% (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9% (28/72)]高于成功组[23.8%(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大<6 cm、胎头位于坐骨棘上及宫口开大8~10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1%(44/72)]高于成功组[38.9%(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2%(52/72)]明显高于成功组[34.1 %(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.  相似文献   

19.
ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly. Issue 4 for 2009 contains 4027 complete reviews, 1906 protocols for reviews in production, and 11447 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 600,000 randomized controlled trials, and 12,200 cited papers in the Cochrane methodology register. The health technology assessment database contains over 7500 citations. This edition of the Library contains 90 new reviews, of which 19 have potential relevance for practitioners in pain and palliative medicine.  相似文献   

20.
ZusammenfassungFragestellung Es wurde geprüft, wie sich der Differenziertheitsgrad zweier Schmerzmessmethoden auf Angaben zur Ausgedehntheit klinischer Schmerzen auswirkt. Zugleich wurde der Referenzzeitraum variiert, über den die Patienten berichten sollten.Methode Erfasst wurde der Einfluss zu Lasten der Befragungsdifferenziertheit durch den Vergleich zweier Körperschema-Bildvorlagen. Drei Referenzzeiträume (Schmerz aktuell, letzte Woche, letztes halbes Jahr) wurden vorgegeben.Ergebnisse Patienten mit ausgedehnten Schmerzen gaben bei differenzierter Befragung um so mehr Schmerzen an, je weiter die Schmerzen zurück lagen und je größer der Berichtszeitraum war. Patienten mit gelenknahen Schmerzen gaben bei hoch differenzierter Befragung weniger ausgedehnte Schmerzen in der Vergangenheit an als bei globaler Einschätzung. Patienten mit Rückenschmerzen berichteten bei differenzierter Befragung zum aktuellen Schmerz über weniger ausgedehnte Schmerzen als bei globaler Befragung.Schlussfolgerung Die Angaben zur Schmerzausdehnung variieren vor allem bei Patienten mit ausgedehnten Schmerzen in Abhängigkeit von der Differenziertheit der Befragung. In diesen Fällen ist die Wahrscheinlichkeit erhöht, dass sich die Beschwerdesymptomatik zumindest teilweise erst in der Reaktion auf die situativen Befragungsbedingungen konstituiert und daher nicht auf andere Befragungsbedingungen generalisiert werden kann.  相似文献   

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