The injury of marginal mandibular branch unexpectedly promotes the repair of buccal branch of facial nerve in a rat model

Conclusions: The results indicate that the injury of the marginal mandibular branch improved the recovery of the buccal branch in a rat model.

Objective: The aim of this study was to investigate whether the injury or intactness of the marginal mandibular branch affects the regeneration of the facial nerve buccal branch in a rat model.

Methods: This experiment was conducted on 30 adult rats, which were randomly and equally divided into two groups. The buccal branch of the facial nerve was transected and reconstructed, with the marginal mandibular branch damaged (group A) or intact (group B). The vibrissae movement of rats was assessed since the 4th week after operation. At the 8th and 12th week, compound muscle action potentials (CMAPs) and morphological changes of injured buccal branches were evaluated.

Results: After the operation, vibrissae movement of rats was eliminated in group A, but it was similar to the health side in group B. CMAPs were recorded from regenerated buccal branches in group A since the 8th week, but no CMAPs could be recorded in group B at each time point. Additionally, the diameter of nerve fibers, the thickness of myelin sheath, and the density of regenerated fibers in group A were significantly larger than those in group B (p?相似文献   

不同浓度地西他滨对破骨细胞分化的影响

目的：探讨不同浓度梯度地西他滨对破骨细胞形成、活性及吸收功能的影响。方法不同浓度地西他滨（0、0.1、0.25和0.5μmol/L）处理单核巨噬（RAW264.7）细胞。通过4’,6?联眯?2?苯基吲哚（4,6?Diamidino?2?phenylindole dihydrochloride, DAPI）染色和微丝绿色荧光探针（F?actin?Trakcer Green）染色后观察F?actin环的形成即破骨细胞轮廓;抗酒石酸酸性磷酸酶检测试剂盒检测细胞上清中的抗酒石酸酸性磷酸酶活性,骨板吸收实验检测破骨细胞的骨吸收能力;Q?PCR实验检测破骨细胞标志基因抗酒石酸酸性磷酸酶（tartrate?resistant acid phosphatase, TRAP）、组织蛋白酶K（cathepsin K, CK）和脊髓基质金属蛋白酶?9（matrix metalloproteinase?9, MMP?9）的mRNA表达。结果不同浓度地西他滨抑制核因子NF?κB配体激活因子（receptor activator of NF?κB ligand, RANKL）诱导RAW264.7细胞形成F?actin环,降低了破骨细胞的TRAP酶活性,抑制了破骨细胞的骨吸收能力,同时也下调了破骨细胞标志基因TRAP、CK和MMP?9的mRNA表达。且随着药物浓度的增高,上述的抑制作用越明显。结论地西他滨抑制破骨细胞形成、活性和骨吸收能力,且这种抑制作用随着药物浓度的增加而逐渐增强。     

骨水泥型人工股骨头置换和PFNA治疗高龄股骨转子间骨折的疗效观察

目的：探讨骨水泥型人工股骨头置换和股骨近端防旋转髓内钉（proximal femoral nail anti?rotation, PFNA）治疗高龄股骨转子间骨折的临床疗效。方法回顾性分析三峡大学人民医院骨科自2008年1月至2013年11月采用骨水泥型人工股骨头置换术和PFNA固定术治疗的高龄股骨转子间骨折患者72例的临床资料,其中人工股骨头置换32例（人工股骨头置换组）,PFNA固定术治疗40例（PFNA组）。比较两组患者的手术时间、术中出血量、术后引流量及关节功能评分等。结果72例患者随访时间为12~36个月,平均20.8个月。PFNA组出现骨折延迟愈合3例。人工股骨头置换组手术时间为（49.1±10.6）min,优于PFNA组的（57.6±7.2）min;术后1个月及末次随访关节功能Harris评分分别为（61.20±4.37）分、（87.92±3.02）分,优于PFNA组的（54.60±3.18）分、（78.86±8.13）分,差异均有统计学意义（均P＜0.05）;而术中出血量、术后引流量分别为（204.2±36.1）ml、（110.5±26.3）ml,多于PFNA组的（98.2±32.3）ml、（58.7±15.7）ml,差异均有统计学意义（P＜0.05）。结论两种方法均可有效治疗高龄股骨转子间骨折,术前根据患者情况选择适当治疗方式均恢复关节功能,提高患者生活质量。     

CMIA与ELISA两种方法在梅毒血清学诊断中的应用

目的:分析化学发光微粒子免疫分析法(CMIA)与酶联免疫吸附法(ELISA)在梅毒血清学诊断中的应用价值。方法:选取104例梅毒患者血清(用TPPA法进行确证)此为试验组,另外选取104例健康人血清,此为对照组,同时用两种方法检测并对结果进行分析。结果:CMIA法检出试验组104例全部为阳性,对照组104例全部为阴性,敏感性及特异性都达到了100%,而ELISA法检出试验组102例阳性2例阴性,对照组103例阴性1例阳性,敏感性98.08%,特异性99.04%。虽然ELISA法敏感性略有差距,但是差异无统计学意义(P>0.05)。结论:两种方法都是理想的梅毒确证试验,敏感性和特异性都较高。CMIA简单方便,可随时检测,单个标本也可操作,需要配套相应的仪器;ELISA需手工操作,繁琐,重复性差,需成批检测,但仪器要求较低,各实验室可根据各自的情况选择。     

Metformin reduces morphine tolerance by inhibiting microglial-mediated neuroinflammation

Background

Tolerance seriously impedes the application of morphine in clinical medicine. Thus, it is necessary to investigate the exact mechanisms and efficient treatment. Microglial activation and neuroinflammation in the spinal cord are thought to play pivotal roles on the genesis and maintaining of morphine tolerance. Activation of adenosine monophosphate-activated kinase (AMPK) has been associated with the inhibition of inflammatory nociception. Metformin, a biguanide class of antidiabetic drugs and activator of AMPK, has a potential anti-inflammatory effect. The present study evaluated the effects and potential mechanisms of metformin in inhibiting microglial activation and alleviating the antinociceptive tolerance of morphine.

Methods

The microglial cell line BV-2 cells and mouse brain-derived endothelial cell line bEnd3 cells were used. Cytokine expression was measured using quantitative polymerase chain reaction. Cell signaling was assayed by western blot and immunohistochemistry. The antinociception and morphine tolerance were assessed in CD-1 mice using tail-flick tests.

Results

We found that morphine-activated BV-2 cells, including the upregulation of p38 mitogen-activated protein kinase (p38 MAPK) phosphorylation, pro-inflammatory cytokines, and Toll-like receptor-4 (TLR-4) mRNA expression, which was inhibited by metformin. Metformin suppressed morphine-induced BV-2 cells activation through increasing AMPK phosphorylation, which was reversed by the AMPK inhibitor compound C. Additionally, in BV-2 cells, morphine did not affect the cell viability and the mRNA expression of anti-inflammatory cytokines. In bEnd3 cells, morphine did not affect the mRNA expression of interleukin-1β (IL-1β), but increased IL-6 and tumor necrosis factor-α (TNF-α) mRNA expression; the effect was inhibited by metformin. Morphine also did not affect the mRNA expression of TLR-4 and chemokine ligand 2 (CCL2). Furthermore, systemic administration of metformin significantly blocked morphine-induced microglial activation in the spinal cord and then attenuated the development of chronic morphine tolerance in mice.

Conclusions

Metformin significantly attenuated morphine antinociceptive tolerance by suppressing morphine-induced microglial activation through increasing AMPK phosphorylation.

     

Influence of far upstream element binding protein 1 gene on chemotherapy sensitivity in human U251 glioblastoma cells

Introduction

The aim of this study was to determine the influence of the far upstream element binding protein 1 gene (FUBP1) on chemotherapy sensitivity in human U251 glioblastoma cells.

Material and methods

Real-time polymerase chain reaction (PCR) was used to determine the expression of the FUBP1 gene in 43 cases of human brain gliomas. Western blot analysis was used to determine the inhibitory effect of RNA interference on FUBP1 gene expression. Methyl thiazolyl tetrazolium assay (MTT) and flow cytometry methods were used to determine the growth inhibitory rate and apoptosis rate of the U251 cells with FUBP1 silencing. The growth inhibitory rate and apoptosis rate were further determined after treatment of those U251 cells with cisplatin (DDP).

Results

The expression of FUBP1 mRNA was up-regulated significantly in gliomas, 177.65% as much as in peri-cancerous tissues (p < 0.05). The expression of FUBP1 protein was inhibited significantly with siRNA-FUBP1 (p < 0.05). In FUBP1-silenced cells, the growth inhibitory rate increased from 1.4% to 29.5%, and the apoptosis rate increased from 2.68% to 5.84% (p < 0.05 for both). After treating with DDP at various concentrations (1, 3, 5 µg/ml), the growth inhibitory rate of FUBP1-silenced cells increased from 14.42%, 17.46% and 23.55% to 21.69%, 27.51% and 37.57%; the apoptosis rate increased from 8.85%, 14.37% and 18.21% to 13.25%, 18.46% and 26.52%.

Conclusions

The up-regulation of FUBP1 relates to the carcinogenesis of gliomas. FUBP1 silencing increases the growth inhibitory rate and apoptosis rate of the U251 cells, and enhances the chemotherapy sensitivity of U251 cells to DDP.     

不同时间点应用甲氨蝶呤对大鼠脊髓损伤后神经细胞凋亡的影响

目的探讨不同时间点应用低剂量甲氨蝶呤(methotrexate,MTX)对大鼠脊髓损伤(spinal cord injury,SCI)后神经细胞凋亡的作用,探讨其潜在的神经保护机制与合适的给药时机。方法取成年雄性SD大鼠120只,体质量247~286 g,随机分为4组(n=30),分别为假手术组(A组)、对照组(B组)、MTX治疗组(C组)和MTX预防组(D组)。A组仅行椎板切除,B、C、D组采用改良Allen法制备SCI模型;C组于术后1、6、12、18、24 h,D组于术前30 min及术后6、12、18、24 h,经尾静脉注射MTX(0.5 mg/kg);A、B组于术后1、6、12、18、24 h注射等量生理盐水。术后观察大鼠一般情况,于1、3、7、14、21 d采用BBB评分进行神经功能评估;取材行组织学观察脊髓形态学变化,免疫组织化学染色观察半胱氨酸天冬氨酸蛋白酶3(Caspase-3)表达,TUNEL标记观察细胞凋亡水平。结果 B、C、D组实验过程中共死亡10只大鼠,最终各组纳入25只大鼠进行观察。各时间点A组BBB评分均高于B、C、D组(P0.05),从3 d开始C、D组评分明显高于B组(P0.05);D组从3 d开始均高于C组,除21 d(P0.05)外,其余各时间点比较差异均有统计学意义(P0.05)。组织学观察示,A组术后各时间点脊髓组织为正常结构;D组SCI程度轻于B、C组,C组轻于B组;从14 d开始,B、C、D组病变范围基本固定。各时间点B、C、D组Caspase-3及TUNEL阳性细胞数均显著多于A组,B组多于C、D组,比较差异有统计学意义(P0.05);C组Caspase-3阳性细胞数均多于D组,其中3、7、14 d比较差异有统计学意义(P0.05),而TUNEL阳性细胞数仅3、7 d显著多于D组(P0.05)。术后1、3、7、14、21 d,B、C、D组组内Caspase-3、TUNEL阳性细胞数均成正相关(P0.05)。结论低剂量MTX能够通过抑制神经细胞凋亡有效阻止SCI的继发性损伤;预防性应用MTX效果优于单纯治疗性应用。