超声内镜引导下钬激光对裸鼠胰腺原位种植瘤消融效果的实验研究

目的：本研究的主要目的是探索超声内镜引导下钬激光对裸鼠胰腺原位种植瘤消融治疗的有效性,为超声内镜引导下钬激光对肿瘤的消融治疗应用于临床提供实验基础。方法：选取4-6周龄的雄性balb/c裸鼠20只,将胰腺癌细胞株SW1990接种至裸鼠胰腺,构建裸鼠原位种植瘤模型,随机分配裸鼠成2组,每组10只,实验组给予超声内镜引导下钬激光的消融治疗,对照组不给予任何治疗。于治疗后第7天,第14天和第28天予超声内镜下测量实验组及对照组裸鼠肿瘤的大小,观察裸鼠活动、食欲等变化。解剖裸鼠,取出肿瘤,行HE染色显微镜下观察胰腺肿瘤组织的病理变化。结果：所有裸鼠均可耐受超声内镜引导下钬激光的消融治疗,实验组裸鼠术后无明显不适反应,精神状态、活动量及食欲较好,对照组裸鼠精神状态逐渐恶化,身材极度消瘦,消融后28天实验组裸鼠肿瘤体积缩小,对照组裸鼠肿瘤增大,两组肿瘤体积的变化差异具有统计学意义,钬激光的治疗疗效大约持续28天左右。HE染色可见消融部位呈凝固性坏死。结论：超声内镜引导下钬激光对裸鼠原位种植瘤的消融治疗是有效的,治疗疗效大约持续28天,主要引起肿瘤组织的凝固性坏死。     

Rho-kinase inhibition improves ischemic perfusion deficit in hyperlipidemic mice

Hyperlipidemia is a major cardiovascular risk factor associated with progressive cerebrovascular dysfunction and diminished collateral perfusion in stroke. Rho-associated kinase (ROCK) may be an important mediator of hyperlipidemic vascular dysfunction. We tested the efficacy of acute or chronic ROCK inhibition on the size of dynamic perfusion defect using laser speckle flowmetry in hyperlipidemic apolipoprotein E knockout mice fed on a high-fat diet for 8 weeks. Mice were studied at an age before the development of flow-limiting atherosclerotic stenoses in aorta and major cervical arteries. Focal ischemia was induced by distal middle cerebral artery occlusion (dMCAO) during optical imaging. The ROCK inhibitor fasudil (10 mg/kg) was administered either as a single dose 1 hour before ischemia onset, or daily for 4 weeks. Fasudil decreased both baseline arterial blood pressure and cerebrovascular resistance (CVR) by ∼15%, and significantly improved tissue perfusion during dMCAO. Interestingly, peri-infarct depolarizations were also reduced. Chronic treatment did not further enhance these benefits compared with acute treatment with a single dose. These data show that ROCK inhibition improves CVR and ischemic tissue perfusion in hyperlipidemic mice.     

Impaired fear extinction in mice lacking protease nexin‐1

The serine protease inhibitor protease‐nexin‐1 (PN‐1) has been shown to modulate N‐methyl‐d ‐aspartate receptor (NMDAR)‐mediated synaptic currents and NMDAR‐dependent long‐term potentiation of synaptic transmission. Here, we analysed the role of PN‐1 in the acquisition and extinction of classical auditory fear conditioning, two distinct forms of learning that both depend on NMDAR activity in the amygdala. Immunostaining revealed that PN‐1 is expressed throughout the amygdala, primarily in γ‐aminobutyric acid containing neurons of the central amygdala and intercalated cell masses (ITCs) and in glia. Fear extinction was severely impaired in mice lacking PN‐1 (PN‐1 KO). Consistent with a role for the basal nucleus of the amygdala in fear extinction, we found that, compared with wild‐type (WT) littermate controls, PN‐1 KO mice exhibited decreased numbers of Fos‐positive neurons in the basal nucleus after extinction. Moreover, immunoblot analysis of laser‐microdissected amygdala sub‐nuclei revealed specific extinction‐induced increases in the level of phosphorylated alpha‐calcium/calmodulin protein kinase II in the medial ITCs and in the lateral subdivision of the central amygdala in WT mice. These responses were altered in PN‐1 KO mice. Together, these data indicate that lack of extinction in PN‐1 KO mice is associated with distinct changes in neuronal activity across the circuitry of the basal and central nuclei and the ITCs, supporting a differential impact on fear extinction of these amygdala substructures. They also suggest a new role for serine protease inhibitors such as PN‐1 in modulating fear conditioning and extinction.     

低强度激光联合照射口咽部和犊鼻穴对实验性膝骨关节炎兔氧自由基代谢的影响

背景：目前激光照射治疗膝骨关节炎以局部照射为主、输出功率和信号较为单一,多次照射易产生生物适应而降低疗效。 目的：探讨携带有针灸补泻信息的低能量激光联合多部位照射对实验性膝骨关节炎兔血清和滑膜组织中氧自由基代谢的影响。 方法：向兔膝关节腔内注入木瓜蛋白酶制备兔膝骨关节炎模型。穴位照射组和联合照射组分别以低能量激光单独照射犊鼻穴或联合照射口咽部及犊鼻穴,10 min /d,5 d为1个疗程,连续3个疗程。参照试剂盒方法测定血清一氧化氮、丙二醛水平和超氧化物歧化酶活性及膝关节滑膜组织丙二醛水平和超氧化物歧化酶活性;苏木精-伊红染色观察膝关节软骨组织形态学变化。 结果与结论：兔膝骨关节炎后,软骨出现明显的病理损伤,血清一氧化氮、丙二醛水平均显著升高(P < 0.01或P < 0.05),超氧化物歧化酶活性显著降低(P < 0.05)。经激光穴位照射和激光联合照射后病理损伤有所减轻,血清一氧化氮、丙二醛水平均下降,超氧化物歧化酶活性升高,以联合照射组效果显著(P < 0.05)。兔膝关节滑膜组织中丙二醛水平和超氧化物歧化酶活性变化与血清中相一致。说明低能量激光联合照射口咽部和犊鼻穴能够纠正膝骨关节炎兔氧自由基代谢紊乱,减轻关节软骨细胞结构损伤,其效果优于激光单独照射犊鼻穴。     

Convergence of entorhinal and CA3 inputs onto pyramidal neurons and interneurons in hippocampal area CA1--an anatomical study in the rat

The entorhinal cortex (EC) conveys information to hippocampal field CA1 either directly by way of projections from principal neurons in layer III, or indirectly by axons from layer II via the dentate gyrus, CA3, and Schaffer collaterals. These two pathways differentially influence activity in CA1, yet conclusive evidence is lacking whether and to what extent they converge onto single CA1 neurons. Presently we studied such convergence. Different neuroanatomical tracers injected into layer III of EC and into CA3, respectively, tagged simultaneously the direct entorhino-hippocampal fibers and the indirect innervation of CA1 neurons by Schaffer collaterals. In slices of fixed brains we intracellularly filled CA1 pyramidal cells and interneurons in stratum lacunosum-moleculare (LM) and stratum radiatum (SR). Sections of these slices were scanned in a confocal laser scanning microscope. 3D-reconstruction was used to determine whether boutons of the labeled input fibers were in contact with the intracellularly filled neurons. We analyzed 12 pyramidal neurons and 21 interneurons. Perforant path innervation to pyramidal neurons in our material was observed to be denser than that from CA3. All pyramidal neurons and 17 of the interneurons received contacts of both perforant pathway and Schaffer input on their dendrites and cell bodies. Four interneurons, which were completely embedded in LM, received only labeled perforant pathway input. Thus, we found convergence of both projection systems on single CA1 pyramidal and interneurons with dendrites that access the layers where perforant pathway fibers and Schaffer collaterals end.     

高精度快速成型模型的制作精度及其体外细胞毒性

背景：快速成型技术在手术模拟与组织工程研究的应用日益广泛,而现场手术模拟和术前指导对模型的精度要求非常高,且在术中使用模型应要求其对人体无毒性。 目的：评估不同后处理方法快速成型模型的成型精度及其细胞毒性。 方法：采用选择性激光烧结技术制作标准圆柱体样品模型,并经浸蜡或树脂处理后,测量模型的高度和底面直径,评估其成型精度。然后采用噻唑蓝法研究不同后处理的模型件对小鼠成纤维细胞L929和小鼠前成骨细胞MC3T3-E1的毒性作用。实验分为石蜡处理组、树脂处理组、阴性对照组(未处理模型)、空白对照组(新鲜培养基)及阳性对照组(体积分数5%的DMSO),培养2 d后,490 nm处测量吸光度,并计算其相对增殖率和评价细胞毒性等级。 结果与结论：浸蜡、树脂处理后的模型以及模型原件的精度为95%~97%,误差为0.5~1 mm。模型浸提液培养L929和MC3T3-E1细胞2 d后,各实验组的细胞相对增殖率均大于80%,石蜡处理后的快速成型模型对L929和MC3T3-E1有较低的毒性作用,而树脂处理和未处理的模型对这两种细胞均无毒性作用,细胞毒性均为0~1级。     

Association study of the CNS patterning genes and autism in Han Chinese in Taiwan

Autism is a complex neurodevelopmental disorder with high heritability. Despite different approaches worldwide to identify susceptibility loci or genes for autism spectrum disorders (ASDs), no consistent result has been reported. CNS patterning genes have been recognized as candidate genes for autism based on neuroimage and neuropathology evidence. This study investigated four candidate genes (WNT2, EN2, SHANK3, and FOXP2) by a tag SNP approach in a family-based association study. The trio samples include 1164 subjects from 393 families, including 393 probands (aged 9.1 ± 4.0 years; male, 88.6%) diagnosed with autistic disorder (n = 373) or Asperger's disorder (n = 20) according to the DSM-IV diagnostic criteria and confirmed by the Chinese ADI-R interview. Three tag SNPs of EN2 (7q36), 6 SNPs of WNT2 (7q31-33), 5 SNPs of SHANK3 (22q13.3), 3 SNPs of FOXP2 (7q31) were genotyped. TDT analysis was done to test the association of each tag SNP and haplotype. There was no association with autism for 17 tag SNPs of WNT2, EN2, SHANK3, and FOXP2 based on SNP analyses. Haplotype analyses did not reveal significant association except for the 6 tag SNPs of WNT2 gene showing a significant association on one haplotype composed of rs2896218 and rs6950765 (G-G) (p = 0.0095). Other haplotypes composed of rs2896218 and rs6950765 (G-G) were also significantly associated with autism. The present study indicates that SHANK3 may not be a critical gene for the etiology of ASDs in Han Chinese population. Inconsistent findings in EN2 and FOXP2 in the Han Chinese population need further clarification. A haplotype of WNT2 (rs2896218-rs6950765: G-G) is significantly associated with ASDs in our trios samples, this finding warrants further validation by different sample and confirmation by functional study.     

Gene expression analyses of neurons,astrocytes, and oligodendrocytes isolated by laser capture microdissection from human brain: Detrimental effects of laboratory humidity

Laser capture microdissection (LCM) is a versatile computer‐assisted dissection method that permits collection of tissue samples with a remarkable level of anatomical resolution. LCM's application to the study of human brain pathology is growing, although it is still relatively underutilized, compared with other areas of research. The present study examined factors that affect the utility of LCM, as performed with an Arcturus Veritas, in the study of gene expression in the human brain using frozen tissue sections. LCM performance was ascertained by determining cell capture efficiency and the quality of RNA extracted from human brain tissue under varying conditions. Among these, the relative humidity of the laboratory where tissue sections are stained, handled, and submitted to LCM had a profound effect on the performance of the instrument and on the quality of RNA extracted from tissue sections. Low relative humidity in the laboratory, i.e., 6–23%, was conducive to little or no degradation of RNA extracted from tissue following staining and fixation and to high capture efficiency by the LCM instrument. LCM settings were optimized as described herein to permit the selective capture of astrocytes, oligodendrocytes, and noradrenergic neurons from tissue sections containing the human locus coeruleus, as determined by the gene expression of cell‐specific markers. With due regard for specific limitations, LCM can be used to evaluate the molecular pathology of individual cell types in post‐mortem human brain. © 2009 Wiley‐Liss, Inc.     

丝裂霉素C在LASEK术矫治中高度近视中的应用

目的:探讨在LASEK治疗中、高度近视术中使用0.2g/L丝裂霉素C预防haze的安全性和有效性。方法:将近视患者120例240眼按左、右眼分为两组,右眼为实验组,左眼为对照组,实验组在完成激光切削后,一次性使用0.2g/L丝裂霉素C液,对照组不用任何药物,术后定期随访角膜刺激症状、角膜上皮生长情况、术后视力、haze等情况。结果:术后1,3,6mo两组术后的刺激症状及角膜上皮愈合时间的差异无统计学意义(P>0.05)。在0级haze中,术后1mo实验组有105眼(87.5%),对照组有62眼(51.7%);术后3mo,实验组有80眼(66.7%),对照组有39眼(32.5%);术后6mo,实验组有96眼(80.0%),对照组有68眼(56.7%),两组差异均有统计学意义(P<0.05)。术后6mo,实验组有91眼(75.8%)的视力大于或等于1.0,对照组有51眼(42.5%)的视力大于或等于1.0,两组的差异有统计学意义(P<0.05)。结论:在LASEK术中使用0.2g/L丝裂霉素C可减轻haze的发生,且安全有效,无明显的毒副作用。