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1.
目的:将猪小肠黏膜下基质与猪髋动脉内皮细胞的复合培养,证实小肠黏膜下基质是否可以成为组织工程化血管的良好载体材料。方法:实验于2005-09/2006-01在同济大学生命科学与技术学院生物医学工程研究所完成了猪小肠黏膜下基质的制备与处理,在复旦大学医学院附属中山医院肿瘤中心完成了猪小肠黏膜下基质的射线照射及消毒,在同济大学生命科学与技术学院实验室完成了猪髋动脉内皮细胞与猪小肠黏膜下基质的复合培养。实验材料:猪小肠黏膜下基质取自屠宰场获取的新鲜家猪空肠,猪髋动脉内皮细胞(PIEC,中国科学院上海细胞生物学研究所细胞库)。实验分组:分为猪髋动脉内皮细胞单独培养组与猪小肠黏膜下基质-猪髋动脉内皮细胞复合培养组。实验方法:①小肠黏膜下基质采用物理和化学方法处理。②猪髋动脉内皮细胞单独培养。③猪小肠黏膜下基质-猪髋动脉内皮细胞复合培养。实验评估:采用倒置相差显微镜观察细胞的黏附和生长,猪小肠黏膜下基质的组织结构;测定单纯培养猪髋动脉内皮细胞和猪小肠黏膜下基质-猪髋动脉内皮细胞复合培养细胞的生长曲线,每日取3孔培养细胞经消化后细胞计数,取其均值,总共8日;以评价猪小肠黏膜下基质与猪髋动脉内皮细胞的细胞相容性。结果:①小肠黏膜下层表面形态:猪小肠黏膜下基质为浅白色薄膜状,厚度约80μm,光镜下可见处理后膜的上下两侧结构不同,一面较粗糙,孔径较大,用于细胞种植,另一面较平坦。单经物理方法处理后的猪小肠黏膜下基质表面有大量残留的细胞碎屑,再行化学处理后基质表面无细胞碎屑残留,胶原纤维未受损。②单独培养与复合培养的猪髋动脉内皮细胞形态对比:倒置显微镜观察,猪小肠黏膜下基质与猪髋动脉内皮细胞复合培养接种细胞24h贴壁,第1~4天时细胞增殖不明显,第5天有较多细胞直接贴附于材料边缘,随时间延长,猪髋动脉内皮细胞在猪小肠黏膜下基质表面黏附生长良好,细胞均匀分布于基质,细胞形态多为梭形及多角形。与猪髋动脉内皮细胞单独培养细胞相比,伪足不明显,细胞形态也相对规则。③细胞生长曲线:复合培养组的细胞计数前4d较单纯培养组少,第5天后时细胞计数高于单纯培养组,复合培养组在接种的后1d数量最少,两组细胞计数均在最后1d达到最大值。结论:猪髋动脉内皮细胞可在猪小肠黏膜下基质上黏附生长,猪小肠黏膜下基质可促进猪髋动脉内皮细胞的增殖,猪小肠黏膜下基质有良好的细胞相容性,其孔径、结构有助于猪髋动脉内皮细胞的黏附和生长,是良好的组织工程生物衍生材料。  相似文献   

2.
目的:观察猪小肠黏膜下层修复兔膝关节内侧副韧带缺损的作用效果。方法:实验于2006-01/08在上海交通大学医学院附属第三人民医院动物房及中心实验室完成。选用新西兰大白兔24只,动物左右两膝分别作为猪小肠黏膜下层移植组及自体韧带移植组,分别制作兔膝关节内侧副韧带缺损模型。猪小肠黏膜下层移植组植入猪小肠黏膜下层,猪小肠黏膜下层作为细胞外基质,无免疫原性;自体韧带移植组植入对侧副韧带。分别于术后2,4,8和12周取白兔膝关节内侧副韧带,行大体形态观察及组织学和力学检查,两组间进行对比观察。结果:24只白兔均进入结果分析,无脱失。①内侧副韧带移植段大体形态观察结果:术后4周,猪小肠黏膜下层移植组有大量的肉芽增生,与周围粘连;自体韧带移植组肉芽组织与周围粘连相对少。术后8周,猪小肠黏膜下层移植组组织梭形增生;自体韧带移植组组织平行增生,直径略小于猪小肠黏膜下层移植组。术后12周,猪小肠黏膜下层移植组增生组织平行于自体韧带,大小一致;自体韧带移植组增生组织直径与猪小肠黏膜下层移植组相一致。②内侧副韧带移植段光镜检查结果:术后2周,猪小肠黏膜下层移植组少量炎性细胞浸润,成纤维细胞数目不多,血管增生明显;自体韧带移植组炎性细胞稀少。术后4周,猪小肠黏膜下层移植组有大量的成纤维细胞,胶原增多;自体韧带移植组成纤维细胞和胶原比猪小肠黏膜下层移植组多。术后8周,猪小肠黏膜下层移植组纤维细胞为主,纤维纵向排列;自体韧带移植组也以成纤维细胞为主。术后12周,两组均有大量的胶原,排列整齐,细胞和纤维密度相一致。③内侧副韧带力学检查结果:术后4周猪小肠黏膜下层移植组白兔内侧副韧带的最大拉伸负荷大于自体韧带移植组[(36.8±8.9,25.3±7.3)N(P<0.05)],术后2,8,12周,两组差异无显著性意义(P>0.05)。结论:猪小肠黏膜下层是一种良好的生物材料,可以用来移植修复膝关节内侧副韧带缺损和损伤,移植后期具有与自体韧带相似的替代修复作用。  相似文献   

3.
背景:小肠黏膜下层具有良好的细胞、组织相容性和降解性,是一种理想的组织工程支架材料,将脂肪基质干细胞与小肠黏膜下层复合后进行定向诱导,可构建靶组织,具有一定的临床应用潜能.目的:制备脱细胞猪小肠黏膜下层基质,并检验其与家兔脂肪基质干细胞的生物相容性.方法:使用酶消化-高盐水脱细胞法处理猪小肠黏膜下层,石蜡切片检测脱细胞效果,扫描电镜观察小肠黏膜下层表面结构.体外分离培养家兔脂肪基质干细胞,分别将第3代脂肪基质干细胞单面复合和双面复合至小肠黏膜下层培养1周观察材料上下表面细胞复合情况.结果与结论:小肠黏膜下层材料为白色半透明膜状物,石蜡切片显示细胞去除彻底,扫描电镜显示小肠黏膜下层黏膜结构紧密,浆膜面纤维结构松散.脂肪基质干细胞与材料复合后,通过相差显微镜观察到细胞在小肠黏膜下层上附着生长,扫描电镜检测提示单面复合脂肪基质干细胞的小肠黏膜下层,仅在上表面有大量细胞生长,下表面无细胞或仅有少量细胞生长,双面复合脂肪基质干细胞的小肠黏膜下层上下表面均可见大量细胞融合生长,石蜡切片可见细胞贴附于材料表面生长.提示酶消化-高渗盐水法可彻底去除小肠黏膜下层表面细胞成分,小肠黏膜下层对脂肪基质干细胞的生长具有良好的支持作用.  相似文献   

4.
背景:国内外研究以大鼠许旺细胞与小肠黏膜下层复合修复神经缺损,取得了较好结果。目的:观察兔许旺细胞与猪小肠黏膜下层体外共培养的生物相容性。方法:采用分步酶消化法分离培养新西兰大白兔许旺细胞,取第3代许旺细胞接种在猪小肠黏膜下层上。结果与结论:①苏木精-伊红染色:复合培养24h后细胞在材料上良好黏附。1周时部分细胞在猪小肠黏膜下层基质层表面呈单层生长,细胞扁平,细胞核呈长梭形,细胞间连接紧密。2周后,细胞呈多层生长。②扫描电镜:复合培养2d,细胞黏附于材料表面并伸展,细胞体呈纺锤形,从胞体伸出两根细长突起,相邻细胞的突起首尾相接连成细胞链或融合或交联或在纤维的侧方平行生长;1周后细胞在材料上大量增殖,呈串珠链黏附于支架上,类似于神经中的Bunger带。表明小肠黏膜下层与许旺细胞有良好的相容性。  相似文献   

5.
背景:国内外研究以大鼠许旺细胞与小肠黏膜下层复合修复神经缺损,取得了较好结果. 目的:观察兔许旺细胞与猪小肠黏膜下层体外共培养的生物相容性. 方法:采用分步酶消化法分离培养新西兰大白兔许旺细胞,取第3代许旺细胞接种在猪小肠黏膜下层上. 结果与结论:①苏木精-伊红染色:复合培养24 h后细胞在材料上良好黏附.1周时部分细胞在猪小肠黏膜下层基质层表面呈单层生长,细胞扁平,细胞核呈长梭形,细胞间连接紧密.2周后,细胞呈多层生长.②扫描电镜:复合培养2 d,细胞黏附于材料表面并伸展,细胞体呈纺锤形,从胞体伸出两根细长突起,相邻细胞的突起首尾相接连成细胞链或融合或交联或在纤维的侧方平行生长;1周后细胞在材料上大量增殖,呈串珠链黏附于支架上,类似于神经中的Bunger带.表明小肠黏膜下层与许旺细胞有良好的相容性.  相似文献   

6.
背景:关节软骨损伤后无论是否施加干预,都难以达到满意的修复效果.目的:观察以猪自体软骨细胞为种子细胞复合脱细胞猪小肠黏膜下层构建组织工程软骨的可行性.方法:将培养至第3代的猪膝关节软骨细胞接种于小肠黏膜下层膜上,复合培养48 h,构建细胞-载体复合物,光学显微镜、扫描电镜观察软骨细胞在小肠黏膜下层膜上的生长情况.结果与结论:苏木精-伊红染色见细胞在小肠黏膜下层基质层表面呈单层或复层生长;免疫组织化学染色结果显示软骨细胞与小肠黏膜下层表面之间形成一条连续阳性表达条带;扫描电镜见软骨细胞在支架孔隙内贴壁良好生长.  相似文献   

7.
背景:关节软骨损伤后无论是否施加干预,都难以达到满意的修复效果。目的:观察以猪自体软骨细胞为种子细胞复合脱细胞猪小肠黏膜下层构建组织工程软骨的可行性。方法:将培养至第3代的猪膝关节软骨细胞接种于小肠黏膜下层膜上,复合培养48h,构建细胞-载体复合物,光学显微镜、扫描电镜观察软骨细胞在小肠黏膜下层膜上的生长情况。结果与结论:苏木精-伊红染色见细胞在小肠黏膜下层基质层表面呈单层或复层生长;免疫组织化学染色结果显示软骨细胞与小肠黏膜下层表面之间形成一条连续阳性表达条带;扫描电镜见软骨细胞在支架孔隙内贴壁良好生长。  相似文献   

8.
目的:观察小肠黏膜下层与半月板纤维软骨细胞的组织相容性,评价其作为组织工程半月板支架材料的可行性及应用价值。方法:实验于2005-06/12在华中科技大学同济医学院附属协和医院骨科实验室完成。①物理和化学方法处理猪小肠黏膜下层支架材料。②体外复合培养兔半月板纤维软骨细胞与小肠黏膜下层。③通过苏木精-伊红染色观察小肠黏膜下层的组织学结构表现;倒置相差显微镜观察细胞生长及与生物材料附着的情况;扫描电镜观察半月板纤维软骨细胞与小肠黏膜下层复合情况。结果:①小肠黏膜下层组织学检查:经物理方法处理后的小肠黏膜下层表面有大量残留的细胞碎屑;化学方法处理后的小肠黏膜下层表面没有残留的细胞碎屑,胶原纤维未受损。②细胞生长及附着情况:相差显微镜观察,复合培养第1~4天时细胞增殖不明显,第5天有较多细胞直接贴附于材料边缘,第7天有大量细胞向材料边缘聚集,细胞形态多为梭形或多角形,连成一片。③细胞与支架复合情况:扫描电镜观察,单纯小肠黏膜下层支架材料可见黏膜表面较粗糙,孔隙较多,大小不一。半月板纤维软骨细胞与小肠黏膜下层复合培养7d可见细胞间通过突起相互连接附着在材料表面,细胞在小肠黏膜下层支架材料上生长、黏附、增殖良好,并分泌大量的细胞外基质成分。结论:小肠黏膜下层具有良好的组织相容性,可以用作组织工程半月板的支架材料。  相似文献   

9.
侯楠  朱力  文科 《中国临床康复》2013,(16):2950-2955
背景:对于解剖层次复杂的细胞外基质,单纯浸泡法则很难达到理想的去细胞效果.目的:观察灌注法去细胞技术制备小肠黏膜全层去细胞外基质的效果. 方法:采用灌注法去细胞技术,经成年雄性新西兰大白兔小肠黏膜上动脉灌注去离子剂后处理获得小肠黏膜全层去细胞外基质.采用MTT法检测小肠黏膜全层去细胞外基质浸提液(实验组)或含体积分数20%小牛血清的DMEM(对照组)与1月龄新西兰大白兔骨髓间充质干细胞共培养后的细胞相对增殖率. 结果与结论:①大体观察:小肠黏膜上动脉经灌注30 min后变得苍白透明,经2 h灌注后肠段变得苍白剔透,清楚可见脉管纹理.②组织学与扫描电镜观察:灌注法构建的小肠黏膜全层去细胞外基质的去细胞程度均匀,胶原纤维未受损,孔隙多,孔隙率为(86.72±2.98)%.③M TT 检测实验:培养2,4,7 d,实验组细胞A值明显高于对照组细胞A值(P〈0.05),且实验组细胞相对增殖率均〉1.表明采用灌注法构建小肠黏膜全层去细胞外基质可操作性强,安全性好,且小肠黏膜全层去细胞外基质对骨髓间充质干细胞无毒性,并有一定的促生长作用.  相似文献   

10.
背景:小肠黏膜下层是一种能够适合于细胞迁移、生长和增殖的良好的支架材料,已被应用于组织工程研究。目的:分析猪小肠黏膜下层作为组织工程肌腱支架的可行性。方法:将24只左腿屈趾肌腱缺损罗曼鸡随机分组,实验组于缺损处植入猪小肠黏膜下层与鸡胚趾深屈肌肌腱细胞复合物,对照组于缺损处植入单纯猪小肠黏膜下层,植入后3,6,9周检测两组植入材料肌腱最大拉伸强度。结果与结论:植入后第3周时,实验组和对照组植入材料肌腱最大拉伸强度差异无显著性意义。植入后第6,9周时,实验组植入材料肌腱最大拉伸强度高于对照组(P<0.05)。表明肌腱细胞-小肠黏膜下层材料复合构建的组织工程肌腱力学强度强于单纯小肠黏膜下层材料修复肌腱,可作为组织工程肌腱支架材料。  相似文献   

11.
Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.  相似文献   

12.
ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD and monthly online. The January 2011 issue (first quarterly DVD for 2011) contains 4515 complete reviews, 1985 protocols for reviews in production, and 13,521 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 641,000 randomized controlled trials, and 14,018 cited papers in the Cochrane methodology register. The health technology assessment database contains over 9300 citations. One hundred and seven new reviews have been published in the last 3 months, of which five have potential relevance for practitioners in pain and palliative medicine.  相似文献   

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Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.  相似文献   

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Ankle sprains are the most common injury of the musculoskeletal system and are associated with significant societal and economic impacts. It has been proven that classical therapeutic strategies may not be effective in preventing recurrent injuries: the recurrence rates reported in the literature can reach 73%. In order to provide an effective rehabilitation solution, a destabilizing orthosis was developed. This device is equipped with a mechanical articulator reproducing the subtalar mechanics and placed under the heel. In this paper, we present the main results of a preliminary clinical study conducted between 2004 and 2007. All subjects included in this study were treated with the abovementioned orthosis during 10 rehabilitation sessions of 30 minutes each. Data show a relatively low recurrence rate of 12% for the overall population. Moreover, it's of primary importance to note that this satisfactory ratio is largely reduced (3% of recurrence rate) for the 29 patients who performed one training session per month after the 10th initial rehabilitation sessions. Hence, the destabilizing orthosis appears to be an effective solution to prevent recurrent ankle sprains. However, joint protection requires long-term and regular training sessions. This result has motivated the development of a similar device allowing patients to perform training sessions at home. Finally, data obtained in this study are promising awaiting the final results of the comparative, multicentric and independent clinical trials currently managed by the Hospices Civils de Lyon.  相似文献   

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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   

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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   

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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   

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