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1.
目的 探讨小干扰RNA(siRNA)载体介导抑制RhoC表达对人结肠癌裸鼠移植瘤的抑制作用.方法 将人结肠癌细胞HT-29接种于裸鼠背部皮下,成功建立裸鼠移植瘤模型;成瘤后将RhoC-siRNA注射入裸鼠移植瘤瘤体,观察RhoC-siRNA对裸鼠移植瘤生长的影响,Western blot检测瘤体中RhoC、基质金属蛋白(MMP-2)和血管内皮生长因子(VEGF)蛋白表达.结果 RhoC-siRNA对结肠癌裸鼠移植瘤的生长有明显抑制作用,瘤体体积重量明显减轻; RhoC-siRNA治疗后瘤体内VEGF和MMP-2蛋白表达明显下降.结论 通过抑制裸鼠结肠癌移植瘤中RhoC的表达,可以抑制肿瘤的生长并下调VEGF和MMP-2的表达水平.  相似文献   

2.
目的 探讨5-氟尿嘧啶(5-FU)和柳氮磺胺吡啶(sulfasalazine, SZ)对裸鼠皮下移植瘤胰腺癌细胞BxPC-3凋亡基因的作用.方法 用RT-PCR和Western blot法分别检测5-FU及SZ单独或联合作用后裸鼠皮下移植瘤凋亡相关基因NF-κB (p65)、bcl-2及cyclinD1 mRNA和蛋白表达水平以及Bax蛋白表达水平的变化.结果 5-FU (7.5和15 mg/kg)和SZ(10和20 mg/kg)单独或联合作用裸鼠皮下移植瘤,其NF-κB (p65) mRNA表达相对含量和蛋白表达水平与各自对照组比较,除SZ组2个亚组外,其余各组均明显降低(P<0.01); bcl-2和cyclinD1 在mRNA和蛋白水平以及Bax蛋白表达量与各自对照组比较差异均有统计学意义(P<0.01); 上述指标经多因素方差分析显示二者间有明显的交互作用.结论 Bax水平上调, bcl-2、cyclinD1及NF-κB (p65)水平下调可能是SZ可协同增强5-FU诱导裸鼠皮下移植瘤胰腺癌细胞BxPC-3凋亡的机理之一.  相似文献   

3.
目的 构建人端粒酶反转录酶(HTERT)启动子调控的NK4基因重组腺病毒表达系统,探讨NK4基因对结肠癌生长、侵袭转移的抑制作用.方法 以Ad HTERTp-NK4重组腺病毒载体感染结肠癌细胞HTC116,分别以RT-PCR方法和Western blot检测细胞内NK4基因mRNA及蛋白表达量:MTT和体外侵袭实验观察NK4对结肠癌细胞增殖及侵袭转移能力的抑制作用.建立结肠癌裸鼠皮下移植瘤模型,观察重组腺病毒Ad HTERTp-NK4对裸鼠结肠癌移植瘤生长的影响.结果 Ad HTERTp-NK4重组腺病毒载体感染HCT116细胞后,HCT116细胞内NK4 mRNA及蛋白表达水平明显增高:Ad HTERTp-NK4重组腺病毒感染后,HCT116细胞生长抑制率和侵袭抑制率分别为47.14%和40.63%,与Ad HTERTp-GFP(空载体)组(2.75%和2.31%)比较,差异均有统计学意义(均P<0.05).局部注射Ad HTERTp-NK4重组腺病毒载体后,结肠癌裸鼠皮下移植瘤模型肿瘤生长受抑,抑瘤率为47.3%,与空载体组(4.6%)比较,差异有统计学意义(P<0.05).结论 HTERT启动子调控NK4基因靶向治疗系统可有效抑制结肠癌的增殖及侵袭转移,在结肠癌的基因治疗具有较好的应用前景.  相似文献   

4.
目的 探讨瘤内注射慢病毒介导的瘦素受体的特异性ObR- siRNA对MCF-7人乳腺癌细胞裸鼠移植瘤生长的影响.方法 建立荷MCF-7人乳腺癌细胞移植瘤裸鼠模型,共30只,随机分为3组:ObR-siRNA慢病毒干预组、NC- siRNA阴性慢病毒对照组和空白对照组,同时在肿瘤部位周围皮下注射重组人瘦素.隔日测量并记录移植瘤的大小,Real-time PCR和Western blotting方法检测ObR的mRNA和蛋白水平的表达,测量肿瘤体积,计算肿瘤抑制率.结果 成功建立了具有高瘦素微环境的敲减ObR基因的荷MCF-7人乳腺癌细胞移植瘤裸鼠模型.ObR- siRNA慢病毒干预组裸鼠移植瘤的体积与NC- siRNA阴性慢病毒对照组和空白对照组差异有统计学意义(P<0.01,P<0.01).ObR- siRNA慢病毒干预组中的ObR的mRNA和蛋白水平的表达明显降低,相对NC- siRNA阴性慢病毒对照组和空白对照组差异有统计学意义(P<0.01,P<0.01),其抑瘤率为88%.结论 利用慢病毒载体成功将ObR-siRNA导入移植瘤内,且能显著抑制MCF-7细胞裸鼠移植瘤的生长,提示瘦素受体有可能成为乳腺癌基因治疗的靶点.  相似文献   

5.
目的 观察组蛋白去乙酰化酶抑制剂-曲古抑菌素(TSA)在体外和体内对胆囊癌细胞和肝外胆管癌细胞HDAC1表达的影响.方法 用TSA作用于胆囊癌细胞系(Mz-ChA-1)和肝外胆管癌细胞系(QBC939、KMBC、OZ),然后用逆转录-聚合酶链反应(RT-PCR)检测HDAC1之mR-NA的表达变化,用Western blot检测其蛋白的表达变化.将这些细胞接种在裸鼠皮下建立胆囊癌和肝外胆管癌裸鼠种植瘤模型,用免疫组织化学方法 观察TSA在体内对裸鼠种植瘤组织中HDAC1蛋白表达的影响.结果 TSA可以减弱胆囊癌细胞系(Mz-ChA-1)和肝外胆管癌细胞系(QBC939、KMBC、OZ)HDAC1 mRNA和蛋白的表达;TSA作用前后的胆囊癌细胞系(Mz-ChA-1)和肝外胆管癌细胞系(KMBC)裸鼠成瘤组织中的各种蛋白的表达均无变化.结论 TSA在体外可以抑制胆囊癌和肝外胆管癌HDAC1的表达;TSA抑制HDAC1表达的作用可能受到体内环境的影响而消失.  相似文献   

6.
目的探讨环氧化酶-2(COX-2)抑制剂塞来昔布对膀胱癌裸鼠皮下移植瘤及移植瘤细胞凋亡的影响.方法6周龄BALB/c裸鼠8只,建立膀胱癌裸鼠皮下移植瘤动物模型,分为2组,实验组每kg食物添加1 g塞来昔布,观察塞来昔布对裸鼠健康状况、体重变化及移植瘤变化的影响,并用TUNEL方法检测其对移植瘤细胞凋亡的影响.结果塞来昔布对裸鼠健康状况及体重变化无明显影响(P>0.05),对移植瘤的生长有抑制作用,肿瘤抑制率为83.7%(P<0.05),对移植瘤细胞的凋亡有促进作用(P<0.05).结论塞来昔布可能通过促进肿瘤细胞凋亡等途径对膀胱肿瘤产生抑制作用,可能成为膀胱肿瘤化学预防及治疗的辅助手段.  相似文献   

7.
腺病毒介导反义c-myc基因抑制人肝癌细胞生长的研究   总被引:3,自引:0,他引:3  
目的观察重组腺病毒介导反义c-myc基因(Ad-ASmyc)对体外培养人肝癌细胞的生长抑制作用和裸鼠体内移植肝肿瘤的治疗效果.方法Ad-ASmyc感染人肝癌细胞系后,观察细胞生长形态变化,通过细胞生长曲线、流式细胞仪分析、逆转录-聚合酶链反应(RT-PCR)、免疫印迹法(Western blot)分析Ad-ASmyc对人肝癌细胞系SMMC-7721和HCC-9204细胞生长、c-myc和bcl-2基因表达的影响;裸鼠皮下移植瘤内注射3种不同剂量Ad-ASmyc(1×109pfu-A组,1×108pfu-B组,1×107pfu-C组)抑制裸鼠肿瘤生长的差异.结果Ad-ASmyc可高效转导人肝癌细胞系SMMC-7721和HCC-9204,抑制细胞生长(抑制率分别为48.72%和56.88%),诱导肝癌细胞G1期阻滞和凋亡,c-myc、bcl-2 RNA及c-myc蛋白水平下降;瘤内注射3种不同剂量Ad-ASmyc均可明显抑制裸鼠皮下移植肿瘤生长,抑制率分别为47.1%、77.3%和82.6%(与对照组比较,P<0.01).结论重组腺病毒介导的反义c-myc基因能够引起人肝癌细胞c-myc、bcl-2 RNA及c-myc蛋白表达下调和细胞生长抑制,瘤内注射Ad-ASmyc治疗裸鼠皮下移植肝癌有效.  相似文献   

8.
目的 研究乳香胶联合吉西他滨对体内、外培养的人胰腺癌BxPc-3细胞的作用及相关机制.方法 四唑氮蓝还原法检测BxPc-3细胞增殖,流式细胞仪检测细胞凋亡(PI单染色法).Western blot检测不同浓度的乳香胶联合吉西他滨(两药联合组)用药后BxPc-3细胞内测核转录因子-κB(NF-κB) p65亚基以及NF-κB抑制蛋白IkB、凋亡调节蛋白Bcl 2和Bax的表达水平的变化.建立人胰腺癌裸鼠皮下移植瘤模型,监测移植瘤体积变化.结果 两药联合组选择乳香胶(40 mg/L)和吉西他滨(10 mg/L)处理72 h后,抑制人胰腺癌细胞株BxPc-3细胞的作用显著优于单独使用(P<0.01);凋亡比例(45.13±4.01)明显高于乳香胶组或吉西他滨组(P<0.01)和对照组(5.07±1.37,P<0.01);乳香胶或两药联合可抑制NF-κB的活性;对人胰腺癌BxPc-3细胞作用48 h后,Bcl-2的蛋白表达明显低于吉西他滨组和对照组(P<0.01).而IkB和Bax蛋白的表达显著高于吉西他滨组和对照组(P<0.01).与对照组比较,乳香胶或两药联合组可显著抑制裸鼠胰腺癌皮下移植瘤生长(P<0.05).结论 乳香胶可通过抑制NF-κB的活性,增加IkB和Bax蛋白表达、抑制Bcl-2的蛋白表达,发挥抗肿瘤作用,并增强吉西他滨的药效.  相似文献   

9.
目的:研究肽酰脯氨基顺反异构酶B(peptidyl-prolyl cis-trans isomerase,PPIB)蛋白对结肠癌细胞成瘤的影响,探讨结肠癌低氧发病的机制。方法:应用RNAi技术,构建下调PPIB蛋白表达的稳转结肠癌细胞株,移植到BALB/c裸鼠皮下,检测PPIB蛋白下调组与对照组的成瘤差异。TUNEL法检测两组移植瘤的细胞凋亡水平。低氧培养结肠癌细胞株,Western印迹法检测PPIB的表达水平与低氧诱导因子1α(hypoxia-inducible factor-1α,HIF-1α)的关系。结果:下调PPIB蛋白表达后,在裸鼠皮下成瘤的能力显著减弱。SW480-si PPIB下调组的移植瘤体积显著小于SW480-NC对照组(P<0.01)。TUNEL检测显示下调组的癌细胞凋亡比例显著高于对照组(P<0.001)。在低氧环境下培养后,结肠癌细胞的PPIB蛋白表达水平随着低氧条件下HIF-1α的上调而升高(P<0.05)。结论:PPIB蛋白在结肠癌中因为低氧环境而表达升高,且在升高后通过抑制癌细胞的凋亡而发挥促进结肠癌形成和进展的作用,是一个极具临床应用价值的防治结肠癌的分子靶点。  相似文献   

10.
目的:探讨蟾毒灵治疗裸鼠结肠癌原位移植瘤的作用及机制。方法 :应用不同剂量蟾毒灵(0.5、1.0、1.5 mg/kg)处理人结肠癌细胞株HCT-116建立的裸鼠原位移植瘤模型,分别用生理盐水(NS)(0.2 mL/只)、5-FU(25 mg/kg)做阴性(NS组)和阳性对照(5-FU组),测量移植瘤的体积,观察蟾毒灵对移植瘤的肿瘤抑制率、裸鼠生存期的影响;HE染色观察移植瘤的坏死程度;TUNEL标记法检测肿瘤细胞的凋亡指数;荧光定量PCR方法检测基因Caspase-3 mRNA的表达,免疫组化法和Western印迹法检测Caspase-3蛋白的表达。结果:蟾毒灵各剂量组和5-FU组裸鼠原位移植瘤体积明显小于NS组(P<0.05);蟾毒灵各组移植瘤的凋亡指数与NS组相比明显增加(P<0.05)。结论:蟾毒灵能够抑制裸鼠人结肠癌原位移植瘤的生长,促进肿瘤细胞凋亡,其机制可能与上调基因Caspase-3表达有关。  相似文献   

11.
Background : We investigated the vasopressor hormone response following mesenteric traction (MT) with hypotension due to prostacyclin (PGI2) release in patients undergoing abdominal surgery with a combined general and epidural anesthesia. Methods : In a prospective, randomized, placebo-controlled study we administered 400 mg ibuprofen (i.v.) in 42 patients scheduled for abdominal surgery. General anesthesia was combined with epidural anesthesia (T4-L1). Before as well as 5, 15, 30, 45, and 90 min after MT we recorded plasma osmolality, hemodynamics and measured 6-keto-PGFlα (stabile metabolite of PGI2), TXB2 (stabile metabolite of thromboxane A2) active renin, and arginine vasopressin (AVP) plasma concentrations by radioimmunoassay. Catecholamine levels were assessed by high-pressure liquid chromatography (HPLC) with electrochemical detection. Results : Following MT, arterial hypotension occurred along with a substantial PGI2 release. This was completely abolished by ibuprofen administration. Although plasma levels of 6-keto-PGF (1133 (708) vs. 60 (3) ng/L, median (median absolute deviation), P=0.0001, placebo vs. ibuprofen) remained significantly elevated, blood pressure was restored within 30 min after MT in the placebo group. At the same point in time plasma concentrations of TXB2 (164 (87) vs. 58 (1) ng/L, P=0.0001), epinephrine (46 (33) vs. 14 (6) ng/L, P=0.001), AVP (41 ± (18) vs. 12 (7) ng/L, P=0.0004), and active renin (27 (12) vs. 12 (4) ng/L, P = 0.001) were significantly higher in placebo-treated patients. Conclusion : Under combined general and epidural anesthesia arterial hypotension following MT due to endogenous PGI2 release is associated with enhanced release of AVP, active renin, epinephrine and thromboxane A2, presumably contributing to hemodynamic stability within 30 min after MT.  相似文献   

12.
Don Dame 《Artificial organs》1996,20(5):613-617
Abstract: Virtually all blood pumps contain some kind of rubbing, sliding, closely moving machinery surfaces that are exposed to the blood being pumped. These valves, internal bearings, magnetic bearing position sensors, and shaft seals cause most of the problems with blood pumps. The original teaspoon pump design prevented the rubbing, sliding machinery surfaces from contacting the blood. However, the hydraulic efficiency was low because the blood was able to "slip around" the rotating impeller so that the blood itself never rotated fast enough to develop adequate pressure. An improved teaspoon blood pump has been designed and tested and has shown acceptable hydraulic performance and low hemolysis potential. The new pump uses a nonrotating "swinging" hose as the pump impeller. The fluid enters the pump through the center of the swinging hose; therefore, there can be no fluid slip between the revolving blood and the revolving impeller. The new pump uses an impeller that is comparable to a flexible garden hose. If the free end of the hose were swung around in a circle like half of a jump rope, the fluid inside the hose would rotate and develop pressure even though the hose impeller itself did not "rotate"; therefore, no rotating shaft seal or internal bearings are required.  相似文献   

13.
Abstract: A variety of protein-bound or hydrophobic substances, accumulating as a result of pathologic conditions such as exogenous or endogenous intoxications, are removed poorly by conventional detoxification methods because of low accessibility (hemodialysis), insufficient adsorption capabilities (hemosorption), low efficiency (peritoneal dialysis), or economic limitations (high-volume plasmapheresis). Combining advantages of existing methods with microspheric technology, a module-based system was designed. Major operating parameters of the latter can be modified to allow for adjustment to individual clinical situations. An extracorporeal blood circuit including a plasmafilter is combined with a secondary high-velocity plasma circuit driven by a centrifugal pump. Different microspheric adsorbers can be combined in one circuit or applied in sequence. Thus, a prolonged treatment can be tailored using specially designed selective adsorber materials. Comparing this system with existing methods (high-flux hemodialysis, molecular adsorbent recycling system), results from our in vitro studies and animal experiments demonstrate the superior efficiency of substance removal.  相似文献   

14.
Background : Our objective was to determine whether administration of propranolol or verapamil modifies the hemodynamic adaptation to continuous positive-pressure ventilation (CPPV), in particular the regional distribution of cardiac output (CO).
Methods : General hemodynamics and regional blood flows assessed by microsphere technique (15 (μm) were recorded in 16 anesthetized pigs during spontaneous breathing (SB) and CPPV with 8 cm H2O end-expiratory pressure (CPPV8) before and after intravenous administration of propranolol (0.3 mg · kg−1 followed by 0.15 mg · kg−1 · h−1, n=8) or verapamil (0.1 mg · kg−1 followed by 0.3 mg · kg−1 · h−1, n=8).
Results : CPPV8 depressed CO by 25% without shifts in its relative distribution with the exception of a noteworthy increase in adrenal perfusion. Propranolol increased arterial blood pressure, and due to a fall in heart rate, CO dropped by 25%. The kidneys and, to a lesser extent, the splanchic region and central nervous system received increased fractions of the remaining CO at the expense of skeletal muscle flow. Similar patterns were seen during SB and CPPV8 such that the combination of propranolol and CPPV8 depressed CO by 50%. The circulatory effects of verapamil were less evident but myocardial perfusion tended to increase.
Conclusions : The combination of propranolol or verapamil with CPPV does not result in any specific hemodynamic interaction in anesthetized pigs, except that the combined effect of propranolol and CPPV may severely reduce CO.  相似文献   

15.
Background : Inhibitory effects of volatile anaesthetics on platelet aggregation have been demonstrated in several studies. However, the influence of volatile anaesthetics on intracoronary platelet adhesion has not been elucidated so far.
Methods : Isolated hearts of guinea pigs were perfused with buffer in the absence or presence of volatile anaesthetics (0.5 and 1 MAC) at constant coronary flow rates of 5 ml/min for 25 min, then 1 ml/min for 30 min and again 5 ml/min for 10 min. Before, during and after low-flow perfusion, a bolus of human platelets was applied into the coronary system. To simulate thrombogenic conditions, 0.3 U/ml human thrombin was infused during low-flow perfusion and reperfusion. The number of platelets sequestered to the endothelium was calculated from the difference between coronary in- and output of platelets. The myocardial production of lactate and consumption of pyruvate and coronary perfusion pressure were also determined.
Results : At a flow rate of 5 ml/min only about 3% of the applied platelets did not emerge from the coronary system, in any group. In contrast, 13.1±1.2% (mean±SEM) of infused platelets became adherent in low-flow perfusion in the control group without anaesthetic. The adherence was reduced with each 1 MAC isoflurane (to 6.2±1.2%), sevoflurane (to 4.4±0.9%) or halothane (to 3.2±1.5%) (each P <0.05 vs. control). Volatile anaesthetic, 0.5 MAC, did not inhibit platelet adhesion to a statistically significant extent in any case. Perfusion pressure and metabolic parameters were not statistically different between the control and the hearts exposed to anaesthetics.
Conclusion : Volatile anaesthetics in a concentration of 1 MAC can reduce the adhesion of platelets in the coronary system under reduced flow conditions. This action does not arise from vasodilation or inhibition of ischaemic stress.  相似文献   

16.
Background: Obesity is increasing globallly, including in the formerly "Eastern Bloc" countries. Methods: A survey was made of obesity and bariatric surgery. Results: In the 8 East and Central European countries studied, with total population 300 million, roughly 43% of the population was overweight (BMI 25-30), 23% obese (BMI > 30), with about 15 million people morbidly obese (BMI > 40). From 0-10 morbidly obese individuals/100,000/year undergo bariatric surgery. Conclusion: Most countries were found to provide inadequate treatment for obesity.The majority of the morbidly obese are not treated effectively. However, health-care awareness of obesity and bariatric surgeons are slowly increasing.  相似文献   

17.
Background: It has been shown that the depressive effects of both propofol and midazolam on consciousness are synergistic with opioids, but the nature of their interactions on other physiological systems, e. g. respiration, has not been fully investigated. The present study examined the effect of propofol and midazolam alone and in combination with fentanyl on phrenic nerve activity (PNA) and whether such interactions are additive or synergistic. Methods: PNA was recorded in 27 anaesthetised and artificially ventilated rabbits. In three groups, propofol, fentanyl and midazolam were administered intravenously in incremental doses to construct dose-response curves for the depressant effects of each one on PNA. In another two groups, the effect of pretreatment with either fentanyl 1 μg · kg?1 i. v. or midazolam 0.05 mg · kg?1 i. v. on the effects of propofol and fentanyl respectively on PNA were studied. Results: Propofol and fentanyl caused a dose-dependent depression of PNA with complete abolition at the highest total doses of 16 mg · kg?1 i. v. and 32 μg · kg?1 i. v., respectively. In contrast, midazolam in incremental doses to a total of 0.8 mg · kg?1 reduced mean PNA by 63%, but approximately 12% of PNA remained at a total dose as high as 6.4 mg · kg?1. The mean ED50s, calculated from dose-response curves, were 5.4 mg · kg?1, 3.9 μg · kg?1 and 0.4 mg · kg?1 for propofol, fentanyl and midazolam, respectively. Initial doses of either fentanyl 1 μg · kg?1 i. v. or midazolam 0.05 mg · kg?1 i. v. acted synergistically with subsequent doses of either propofol or fentanyl to abolish PNA at total doses of 8 mg · kg?1 and 8 μg · kg?1, respectively. Conclusion: Fentanyl has a synergistic interaction with both propofol and midazolam on PNA and hence potentially on respiration.  相似文献   

18.
Background: Catecholaminergic support is often used to improve haemodynamics in patients undergoing major abdominal surgery. Dopexamine is a synthetic vasoactive catecholamine with beneficial microcirculatory properties. Methods: The influence of perioperative administration of dopexamine on cardiorespiratory data and important regulators of macro- and microcirculation were studied in 30 patients undergoing Whipple pancreaticduodenectomy. The patients received randomized and blinded either 2 μg · kg?1 · min?1 of dopexamine (n=15) or placebo (n=15, control group). The infusion was started after induction of anaesthesia and continued until the morning of the first postoperative day. Endothelin-1 (ET-1), vasopressin, atrial natriuretic peptide (ANP), and catecholamine plasma levels were measured from arterial blood samples. Measurements were carried out after induction of anaesthesia, 2 h after onset of surgery, at the end of surgery, 2 h after surgery, and on the morning of the first postoperative day. Results: Cardiac index (CI) increased significantly in the dopexamine group (from 2.61±0.41 to 4.57±0.78 1 · min?1 · m?2) and remained elevated until the morning of the first postoperative day. Oxygen delivery index (DO2I) and oxygen consumption index (VO2I) were also significantly increased in the dopexamine group (DO2I: from 416±91 to 717±110 ml/m2 · m2; VO2I: from 98±25 to 157±22 ml/m2 · m2), being significantly higher than in the control group. pHi remained stable only in the dopexamine patients, indicating adequate splanchnic perfusion. Vasopressive regulators of circulation increased significantly only in the untreated control patients (vasopressin: from 4.37±1.1 to 35.9±12.1 pg/ml; ET-1: from 2.88±0.91 to 6.91±1.20 pg/ml). Conclusion: Patients undergoing major abdominal surgery may profit from prophylactic perioperative administration of dopexamine hydrochloride in the form of improved haemodynamics and oxygenation as well as beneficial influence on important regulators of organ blood flow.  相似文献   

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Abstract: Photopheresis is a technique in which peripheral blood mononuclear cells, in the presence of a photoacti-vatable compound, are exposed extracorporeally to ultraviolet A light and reinfused, inducing a host autoregula-tory immune response. Experimental work and ongoing clinical studies are helping to define the role of this novel, safe, and non-toxic immunomodulating technology in the field of transplantation.  相似文献   

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