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目的:利用RNA干扰技术抑制大鼠成骨细胞核激活因子受体配体(Ligand of receptor activator of NF-κB,RANKL)的表达,观察成骨细胞RANKL/骨保护素(osteoprotegerin,OPG)比值变化,探讨成骨细胞与破骨细胞生成的相关性。方法:实验于2006-05/10在四川大学华西医院移植免疫实验室(国家重点实验室)完成。①实验材料:体外化学合成4条RANKL序列特异性小干扰RNA(small interfering RNA,siRNA),用Lipofectin2000TM转染成骨细胞。②实验方法:采用荧光实时定量聚合酶链反应检测RANKL mRNA的表达,筛选出最有效的干扰序列。③实验分组:分为最有效的siRNA转染组、阴性对照组和空白对照组,每组设5个平行样本。采用免疫组织化学染色检测成骨细胞中RANKL、OPG蛋白的表达,观察RANKL/OPG比值变化。④实验评估:采用成骨细胞、破骨细胞共培养技术观察以上3组转染后的成骨细胞对破骨细胞生成的影响。结果:①4种siRNA敲除RANKL基因后mRNA的表达:4种siRNA转染成骨细胞后,相对空白对照组,siRNA4分子对RANKLmRNA的抑制作用最为明显,达到89%。siRNA1,siRNA3对RANKLmRNA的抑制作用大于50%,而siRNA2对RANKLmRNA的也有相当的抑制作用。②各组成骨细胞中RANKL和OPG蛋白水平:RANKL和OPG的阳性表达均显示为棕黄色颗粒,RANKL主要分布于成骨细胞的胞浆和胞膜,OPG主要分布于成骨细胞的胞浆。siRNA4转染组RANKL表达(平均积分光密度值)低于空白对照组(分别为3.05±2.17,11.31±1.26),差异有显著性意义(P<0.01);阴性对照组与空白对照相比,差异无显著性意义(P>0.05)。siRNA4转染组、阴性对照组OPG表达与空白对照组比较,差异均无显著性意义(P>0.05)siRNA4转染组RANKL/OPG比值低于空白对照组(分别为0.12±0.03,0.45±0.04),差异有显著性意义(P<0.01)。③转染的成骨细胞对破骨细胞生成的影响:各组共培养形成的破骨细胞形态与分离的成熟破骨细胞相似,为不规则形,胞浆红染,并有伪足形成。siRNA4转染组破骨细胞形成低于空白对照组、阴性对照组[分别为(12±2),(31±7),(28±5)个/孔],差异有显著性意义(P<0.01);阴性对照组与空白对照相比,差异无显著性意义(P>0.05)。结论:RNA干扰沉默RANKL表达可下调成骨细胞RANKL/OPG比值,抑制破骨细胞的生成。 相似文献
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OBJECTIVES: Life-threatening toxic side-effects following 5-FU exposure have been related to deficiency of dihydropyrimidine dehydrogenase (DPD), the rate-limiting enzyme in its catabolism. We presently report a new DPYD gene SNP in a Spanish woman who died from multivisceral 5-FU-induced toxicity. DESIGN AND METHODS: We looked for 22 known SNPs by Pyrosequecing. Then, we sequenced the whole 23 exons of DPYD, along with adjacent intronic sequences. PCR was carried out to determine whether or not exons were deleted in the DPYD. To determine whether the predicted stop codon indeed resulted in a truncated protein, a bacterial expression vector was employed to generate the predicted protein. 500 patients were genotyped to determine allele frequency. RESULTS: A novel mutation 464 T>A was identified in DPYD gene exon 5, resulting in the replacement of leucine 155 by a stop codon in the protein. We confirmed this mutation by Pyrosequencing and its involvement by a protein truncation test. We genotyped the patient's family and the allele frequency was 0.2%. CONCLUSION: The involvement of this variant in 5-FU life-threatening toxicity supports its inclusion in pretherapeutic genetic screening. 相似文献
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Outpatient total laparoscopic hysterectomy 总被引:2,自引:0,他引:2
Thiel J Gamelin A 《The Journal of the American Association of Gynecologic Laparoscopists》2003,10(4):481-483
STUDY OBJECTIVE: To assess the safety of and patient satisfaction with total laparoscopic hysterectomy as an outpatient procedure. DESIGN: Retrospective case study (Canadian Task Force classification III). SETTING: Tertiary care hospital. PATIENTS: Sixty-six consecutive women. INTERVENTION: Outpatient total laparoscopic hysterectomy. MEASUREMENTS AND MAIN RESULTS: Of 66 procedures completed, 6 patients (7.5%) required overnight hospitalization, with 4 of them discharged the next day. Seven (11%) minor postoperative complications occurred. One patient required hospitalization to receive intravenous antibiotics and one for drainage of a cuff hematoma. There were three (4.5%) minor intraoperative complications. One woman developed Clostridium difficile diarrhea as well as deep vein thrombosis and pulmonary embolus. Sixty-three women (95%) were satisfied with the procedure and would recommend it to others. CONCLUSION: Outpatient total laparoscopic hysterectomy is well tolerated, safe, and cost effective. 相似文献
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A phase 1 study of AMG 900, an orally administered pan‐aurora kinase inhibitor,in adult patients with acute myeloid leukemia 下载免费PDF全文
Hagop M. Kantarjian Michael W. Schuster Nitin Jain Anjali Advani Elias Jabbour Erick Gamelin Erik Rasmussen Gloria Juan Abraham Anderson Vincent F. Chow Gregory Friberg Florian D. Vogl Mikkael A. Sekeres 《American journal of hematology》2017,92(7):660-667
Aurora kinases are involved in the pathophysiology of several cancers including acute myeloid leukemia (AML). In this phase 1 study, we investigated the safety and efficacy of AMG 900, an orally administered, highly potent, selective, small‐molecule inhibitor of both Aurora kinase A and B, in patients with AML . Patients with pathologically documented AML who either declined standard treatments or had relapsed from or were refractory to previous therapies were enrolled. Two every‐2‐week dose‐escalation schedules using a modified 3 + 3 + 3 design were evaluated AMG 900 given daily for 4 days with 10 days off (4/10 schedule), and AMG 900 given daily for 7 days with 7 days off (7/7 schedule). Thirty‐five patients were enrolled at 9 different dose levels: 22 patients on the 4/10 schedule (doses from 15 to 100 mg daily), and 13 patients on the 7/7 schedule (doses from 30 to 50 mg daily). Both schedules were tolerated; nausea (31%), diarrhea (29%), febrile neutropenia (29%), and fatigue (23%) were the most common treatment‐related adverse events. Three patients (9%) achieved complete response with incomplete count recovery. Patients with higher baseline expression of a set of specific pathway‐related genes (BIRC5, AURKA, TTK, CDC2, and CCNB1) were more likely to respond in an exploratory biomarker analysis. AMG 900 was tolerated in a general AML population, and pathway‐specific biomarkers identified a potential target population. Future research efforts will be directed toward further exploration of biomarkers of response and combination of AMG 900 with other anticancer agents. 相似文献
68.
Gamelin FX Baquet G Berthoin S Bosquet L 《International journal of sports medicine》2008,29(2):134-138
Intervals between two consecutive cardiac beats (R-R intervals) and the subsequent analysis of heart rate variability (HRV) obtained simultaneously from the Polar S810 heart rate monitor (HRM) and an electrocardiogram (ECG) in a supine position were compared in twelve children (age 9.6 +/- 0.9 years) before and after protocol correction. R-R intervals were significantly different between the ECG and the HRM uncorrected and corrected signal (p < 0.001, effect size [ES] = 0.005, and 0.005, respectively). However, the bias (95 % confidence interval) was 0.80 (- 124.76 - 123.16) ms and 0.80 (- 12.76 - 11.16) ms, respectively. HRV parameters derived from both signals were not different (p > 0.05) and well correlated (r > 0.99, p < 0.05), except SD2 (p < 0.05, ES = 0.000; r = 0.99). These data support the validity of the Polar S810 HRM to measure R-R intervals and make the subsequent HRV analysis in a supine position in children. 相似文献
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Rückert SE 《中国介入心脏病学杂志》2003,11(5):237
一、背景分叉病变血管成形术一直是个困扰医学界的难题。没有现成的严格治疗标准可以依据。大多数技术不是复杂耗时就是成功率低 ,且伴有早期和后期并发症。二、目标我们对因血管分叉病变 (侧支血管直径 >2mm)进行支架置入术的情况进行了注册研究。所用支架类型具有最佳的柔软性 ,较小外径、较小的横梁宽度和较大的侧孔 ,适于直接扩张 ,在不弯曲远端支架末端的情况下容易进入侧支血管 ,大多数情形下无需双导丝技术和对吻球囊。三、方法以调查问卷的形式收集了自 2 0 0 1年 6月 1日起在所有侧支血管直径 >2mm的分叉病变中使用Coroflex 支架… 相似文献