大鼠骨癌痛-慢性吗啡耐受模型的建立

目的 建立大鼠骨癌痛-慢性吗啡耐受模型.方法 鞘内置管成功的成年雌性SD大鼠36只,体重180～200 g,采用随机数字表法,将大鼠随机分为3组(n=12):假手术组(S组)、慢性吗啡耐受组(M组)和骨癌痛+慢性吗啡耐受组(BM组).BM组右侧胫骨上段骨髓腔注入Walker256癌细胞10 μl(4×102个细胞/μl)制备骨癌痛模型,M组注射热灭活的Walker256癌细胞10μl,接种后10 d开始鞘内注射吗啡20μg/kg,2次/d,连续9 d.S组仅暴露右侧胫骨上段.分别于接种Walker256癌细胞前、接种后1、3、6、9 d、给予吗啡1、3、5、7、9 d时测定机械缩足阈值(MWT)和机械缩足持续时间(MWD),并于接种后9 d时行放射学检查,进行骨质破坏评分.最后1次测定痛周后,对右侧足底进行触摸刺激,停止刺激后3 h时取脊髓L4-6节段,测定脊髓背角Foa表达水平.结果 与S组比较,M组MWT降低,MWD延长,脊髓背角Fos表达上调(P＜0.05或0.01);与M组比较,BM组MWT降低,骨质破坏评分升高,MWD延长,脊髓背角Fos表达上调(P＜0.05或0.01).结论 成功制备了大鼠骨癌痛-慢性吗啡耐受模型.

Abstract：

Objective To establish a rat model of bone cancer pain-chronic morphine tolerance. Methods Thirty-six adult female Sprague-Dawley rats weighing 180-200 g in which intrathecal catheters were successfully placed without complications were randomly divided into 3 groups ( n = 12 each) :group sham operation (group S),group chronic morphine tolerance (group M) and group bone cancer pain + chronic morphine tolerance (group BM). Bone cancer pain was induced by intra-tibia inoculation of Walker256 mammary gland carcinoma cells (4 ×102 cells/μl) in group BM, while in group M heat-inactivated Walker256 mammary gland carcinoma cells were given instead, and then 10 days later, intrathecal morphine 20 μg,/kg was administered twice a day for 9 consecutive days. The mechanical paw withdrawal threshold (MWT) and mechanical paw withdrawal duration (MWD) were measured before inoculation, at day 1, 3, 6 and 9 after inoculation, and at day 1, 3, 5, 7 and 9 of morphine administration. The degree of bone destruction was assessed by radiological analysis at day 9 after inoculation. After the last measurement of pain threshold, the rats were given innoxious touch-stimulus. The rats were sacrificed 3 h after stopping the stimulus, and L4-6 segment of the spinal cord was isolated to determine the expression of Fos protein in the spinal dorsal horn. Results Compared with group S, MWT was significantly decreased, MWD was significantly prolonged and the expression of Fos protein was up-regulated in group M ( P ＜ 0.05 or 0.01 ). MWT was significantly decreased, MWD was significantly prolonged, bone destruction scores were significantly increased,and the expression of Fos protein was up-regulated in group BM compared with group M ( P ＜ 0.05 or 0.01 ). Conclusion A rat model of bone cancer pain-chronic morphine tolerance is successfully established.     

携带DREAM基因小分子干扰RNA重组质粒的构建☆

背景：DREAM是一种多功能蛋白,在细胞中不同位置与不同靶蛋白结合,体外细胞培养和动物实验均证明DREAM参与了许多疾病的发病机制。 目的：构建携带DREAM基因的小分子干扰RNA重组质粒。 方法：设计并合成shRNA对应的两条互补的寡核苷酸链,pDC316-EGFP-U6质粒经BamHⅠ和HindⅢ双酶切与退火后的寡核苷酸连接,转化感受态E.coli DH5α,获得阳性克隆进行PCR和测序鉴定。 结果与结论：经PCR、酶切及测序证实,重组质粒pDC316-EGFP-DREAM-shRNA-U6片段大小为473 bp,其中插入的片断序列和位点与预期完全一致,说明pDC316-EGFP-DREAM-shRNA-U6重组质粒构建成功。     

超声引导臀下和臀横纹下入路坐骨神经阻滞的比较

目的 比较臀下入路和臀横纹下入路超声引导坐骨神经阻滞的效果.方法 选择择期下肢手术患者148例,随机分为臀下入路和臀横纹下入路组,每组74例,在超声联合神经刺激器引导下行坐骨神经阻滞,局麻药为0.5%罗哌卡因20 ml,测定坐骨神经至皮肤距离深度,对比操作时间和调整穿刺方向次数,评价坐骨神经运动阻滞率及主要分支胫神经、腓浅神经、腓肠神经和股后皮神经感觉阻滞效果,记录麻醉相关并发症.根据手术麻醉需要所有病例同时以0.5%罗哌卡因20 ml行腰丛神经阻滞.结果 臀横纹下入路组穿刺时间和调整穿刺针方向次数少于臀下入路组,臀横纹下入路组注药15 min后脚踝运动阻滞率低于臀下入路组,两组间注药30 min后感觉和运动阻滞率无差异,两组手术麻醉效果、镇痛时间和运动阻滞时间无差异.结论 臀下和臀横纹入路坐骨神经阻滞均可用于下肢手术麻醉,臀横纹下入路组操作更方便,可作为高位坐骨阻滞的首选入路.

Abstract：

Objective To compare the effects of subgluteal(SG) and sub-subgluteal-fold(SSGF)approach for ultrasound-guided siatic nerve block. Methods One hundred forty-eight patients undergoing lower limb surgery were randomly divided into two groups to receive SG approaches and SSGF approaches to sciatic nerve block under real time ultrasound guidance. A combined posterior lumbar plexus block under ultrasound guidance was performed for sufficient surgery anesthesia. 20 ml of 0. 5% ropivacaine was used for sciatic nerve and lumbar plexus block separately. Measurements included skin-to-nerve distance,reorientation of the needle during block and execution time,rates of sensory and motor blockade after 15 min and 30 min of injection, quality of surgery blockade, duration of the sensory and motor block, and postoperative complications related to sciatic nerve block. Results In SSGF group, execution time and reorientation of needle for sciatic nerve block was significantly less than those of the SG group( P ＜0.01).But motor blockade in the SG group was quicker when compared with SSGF group ( P ＜0.01). There were no significant differences in the quality and duration of blockade between the two groups. Conclusions Both SG and SSGF approach can be used for sciatic nerve block with equal sensory and motor block rate,whereas sciatic nerve block via SSGF approach was faster and easy to perform than the SG one.     

Inhibition of glial activation in rostral ventromedial medulla attenuates mechanical allodynia in a rat model of cancer-induced bone pain

Descending nociceptive modulation from the supraspinal structures plays an important role in cancer-induced bone pain (CIBP). Rostral ventromedial medulla (RVM) is a critical compo-nent of descending nociceptive facilitation circuitry, but so far the mechanisms are poorly known. In this study, we investigated the role of RVM glial activation in the descending nociceptive facilitation circuitry in a CIBP rat model. CIBP rats showed significant activation of microglia and astrocytes, and also up-regulation of phosphorylated p38 mitogen-activated protein kinase (p38 MAPK) and pro-inflammatory mediators released by glial cells (IL-1β, IL-6, TNF-α and brain-derived neurotro-phic factor) in the RVM. Stereotaxic microinjection of the glial inhibitors (minocycline and ?uorocitrate) into CIBP rats’ RVM could reverse the glial activation and significantly attenuate me-chanical allodynia in a time-dependent manner. RVM microinjection of p38 MAPK inhibitor (SB203580) abolished the activation of microglia, reversed the associated up-regulation of pro-inflammatory mediators and significantly attenuated mechanical allodynia. Taken together, these results suggest that RVM glial activation is involved in the pathogenesis of CIBP. RVM microglial p38 MAPK signaling pathway is activated and leads to the release of downstream pro-inflammatory mediators, which contribute to the descending facilitation of CIBP.     

不同药物治疗全麻患者苏醒期躁动的疗效比较

目的 比较芬太尼,曲马多、布托啡诺和帕瑞昔布对全麻患者苏醒期躁动的治疗作用,探讨患者术后躁动治疗的合理化用药方案.方法 全麻后出现苏醒期躁动的ASA Ⅰ或Ⅱ级成年患者120例,随机均分为四组:芬太尼组(F组)、曲马多组(T组)、布托啡诺组(B组)和帕瑞昔布组(P组),分别给予静脉注射芬太尼1 μg/kg、曲马多1 mg/kg、布托啡诺20 μg/kg和帕瑞昔布40 mg进行治疗.各组患者分别在用药前后进行VAS评分、Prince-Henry疼痛评分、Ramsay镇静评分和RSS躁动评分以评价药物治疗效果,记录患者可能的术后躁动诱发因素和麻醉后恢复室(PACU)停留时间.结果 疼痛与导尿管刺激是引起苏醒期躁动的主要原因,T组躁动缓解率低于其他三组(P＜0.05).与用药前比较,四组患者用药后VAS评分和Prince-Henry疼痛评分均降低(P＜0.05),而用药后T组VAS评分高于其他三组(P＜0.05),用药后四组患者的Ramsay镇静评分较用药前均明显升高,其中B组患者高于其他三组(P＜0.05).B组患者在PACU停留时间长于其他三组(P＜0.05).结论 帕瑞昔布是治疗苏醒期躁动较为安全有效的药物,芬太尼有可能会导致患者发生一过性呼吸抑制,布托啡诺町延长患者在PACU的停留时间,而曲马多对苏醒期躁动的治疗效果欠佳.