外源性p53对Wnt通路抑制因子Dickkopf-1的表达作用

目的研究外源性p53对W nt通路抑制因子D ickkopf-1(DKK-1)的表达作用。方法将携带p53基因的复制缺陷型腺病毒载体(Adp53)导入到p53完全缺失的人肝癌细胞株Hep3B中,并以p53反以寡核苷酸阻断p53阳性细胞p53的表达,以W nt通路的关键因子-βcaten in的改变为功能指标评价W nt通路的变化。以RT-PCR技术检测W nt通路抑制因子DKK-1表达的调节作用,以流式细胞术检测Adp53的转基因情况,肝癌细胞中p53和β-caten in的表达。结果DKK-1mRNA水平在转染Adp53 20 h后即有明显升高,32 h达最高水平,随后逐渐降低。β-caten in表达水平随着转染时间和转染剂量的增加,阳性细胞百分比强度和平均荧光量强度表达水平逐渐下降。以p53反以寡核苷酸阻断p53阳性细胞p53表达后,抑制剂DKK-1的表达逐渐降低,-βcaten in表达水平逐渐增加。结论外源性p53能明显诱导W nt通路抑制剂DKK-1的表达,进而产生抑制W nt通路的作用。     

滋肾清肝代平方中7种成分血药浓度的测定及其在大鼠体内的药代动力学分析

目的:建立UPLC-MS-MS同时测定大鼠血浆中2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷,1-脱氧野尻霉素和决明子苷、红镰霉素龙胆二糖苷、橙黄决明素、甲基钝叶决明素、白藜芦醇的分析方法,研究大鼠服用滋肾清肝代平方后的药代动力学特征。方法:采用Waters Acquity UPLC BEH C18色谱柱,流动相0.1%甲酸水溶液-乙腈梯度洗脱,采用电喷雾电离源,扫描方式为多反应离子监测。结果:7种成分在大鼠血浆中线性关系良好,提取回收率94.83%~106.58%,日内、日间精密度和准确度良好。7种成分在模型组大鼠中的药时曲线下面积、末端半衰期、达峰时间、清除率、表观分布容积、药峰浓度分别为(326.65±26.66)μg·L-1·h-1,(3.64±1.69)h,0.33 h,(60.56±5.32)L·h-1·kg-1,(325.13±167.18)L·kg-1,(169.25±18.02)μg·L-1。结论:该方法特异、快速、准确、灵敏,可用于滋肾清肝代平方在大鼠体内的药动学研究。     

Selective inhibition of JNK with a peptide inhibitor attenuates pain hypersensitivity and tumor growth in a mouse skin cancer pain model

Cancer pain significantly affects the quality of cancer patients, and current treatments for this pain are limited. C-Jun N-terminal kinase (JNK) has been implicated in tumor growth and neuropathic pain sensitization. We investigated the role of JNK in cancer pain and tumor growth in a skin cancer pain model. Injection of luciferase-transfected B16-Fluc melanoma cells into a hindpaw of mouse induced robust tumor growth, as indicated by increase in paw volume and fluorescence intensity. Pain hypersensitivity in this model developed rapidly (< 5 days) and reached a peak in 2 weeks, and was characterized by mechanical allodynia and heat hyperalgesia. Tumor growth was associated with JNK activation in tumor mass, dorsal root ganglion (DRG), and spinal cord and a peripheral neuropathy, such as loss of nerve fibers in the hindpaw skin and induction of ATF-3 expression in DRG neurons. Repeated systemic injections of D-JNKI-1 (6 mg/kg, i.p.), a selective and cell-permeable peptide inhibitor of JNK, produced an accumulative inhibition of mechanical allodynia and heat hyperalgesia. A bolus spinal injection of D-JNKI-1 also inhibited mechanical allodynia. Further, JNK inhibition suppressed tumor growth in vivo and melanoma cell proliferation in vitro. In contrast, repeated injections of morphine (5 mg/kg), a commonly used analgesic for terminal cancer, produced analgesic tolerance after 1 day and did not inhibit tumor growth. Our data reveal a marked peripheral neuropathy in this skin cancer model and important roles of the JNK pathway in cancer pain development and tumor growth. JNK inhibitors such as D-JNKI-1 may be used to treat cancer pain.     

Percutaneous cholecystography in children

Percutaneous cholecystography was performed on 13 children who had biliary system abnormalities: two had biliary hypoplasia, five had sclerosing cholangitis, three had cirrhosis, two had distal choledochal obstruction, and one had an obstructed portoenterostomy. In 12 patients transcholecystic cholangiography showed, without significant complications, the intra-and extrahepatic bile ducts. In one patient with primary sclerosing cholangitis, the intrahepatic bile ducts were not opacified satisfactorily; dilatation of the gallbladder required surgical drainage. The transcholecystic technique is indicated when the intrahepatic bile ducts are either mildly dilated or not dilated.     

The effects of manual therapy and exercise directed at the cervical spine on pain and pressure pain sensitivity in patients with myofascial temporomandibular disorders

Summary No studies have investigated the effects of the treatments directed at the cervical spine in patients with temporomandibular disorders (TMD). Our aim was to investigate the effects of joint mobilization and exercise directed at the cervical spine on pain intensity and pressure pain sensitivity in the muscles of mastication in patients with TMD. Nineteen patients (14 females), aged 19–57 years, with myofascial TMD were included. All patients received a total of 10 treatment session over a 5‐week period (twice per week). Treatment included manual therapy techniques and exercise directed at the cervical spine. Outcome measures included bilateral pressure pain threshold (PPT) levels over the masseter and temporalis muscles, active pain‐free mouth opening (mm) and pain (Visual Analogue Scale) and were all assessed pre‐intervention, 48 h after the last treatment (post‐intervention) and at 12‐week follow‐up period. Mixed‐model anovas were used to examine the effects of the intervention on each outcome measure. Within‐group effect sizes were calculated in order to assess clinical effect. The 2 × 3 mixed model anova revealed significant effect for time (F = 77·8; P < 0·001) but not for side (F = 0·2; P = 0·7) for changes in PPT over the masseter muscle and over the temporalis muscle (time: F = 66·8; P < 0·001; side: F = 0·07; P = 0·8). Post hoc revealed significant differences between pre‐intervention and both post‐intervention and follow‐up periods (P < 0·001) but not between post‐intervention and follow‐up period (P = 0·9) for both muscles. Within‐group effect sizes were large (d > 1·0) for both follow‐up periods in both muscles. The anova found a significant effect for time (F = 78·6; P < 0·001) for changes in pain intensity and active pain‐free mouth opening (F = 17·1; P < 0·001). Significant differences were found between pre‐intervention and both post‐intervention and follow‐up periods (P < 0·001) but not between the post‐intervention and follow‐up period (P > 0·7). Within‐group effect sizes were large (d > 0·8) for both post‐intervention and follow‐up periods. The application of treatment directed at the cervical spine may be beneficial in decreasing pain intensity, increasing PPTs over the masticatory muscles and an increasing pain‐free mouth opening in patients with myofascial TMD.     

White-matter lesions in MR imaging of clinically healthy brains of elderly subjects: possible pathologic basis

Patchy white-matter lesions occur in the magnetic resonance (MR) imaging brain studies of 20%-30% of neurologically healthy elderly subjects. To determine the frequency of histologically verifiable white-matter lesions at autopsy in such subjects the authors examined serial, microscopic, whole brain sections from 15 clinically healthy subjects aged 52-72 years. Small white-matter lesions were found in 12. In these 12, zones of atrophic perivascular demyelination were present in eight brains. These are not the familiar thrombotic, embolic, or ischemic vascular lesions that produce acute necrosis. This mild vascular insufficiency produces atrophy, which has been recognized in the pathology literature but whose clinical significance remains unknown. Other lesions seen were small vascular malformations in the centrum ovale in four brains, diverticula of the lateral ventricle extending into the white matter in three, and an isolated central white-matter infarction in one. All of these lesions are probably the basis of the patchy white-matter lesions seen on MR imaging studies in the neurologically healthy elderly population.