Guided growth of neurons and glia using microfabricated patterns of parylene-C on a SiO2 background

This paper describes a simple technique for the patterning of glia and neurons. The integration of neuronal patterning to Multi-Electrode Arrays (MEAs), planar patch clamp and silicon based ‘lab on a chip’ technologies necessitates the development of a microfabrication-compatible method, which will be reliable and easy to implement. In this study a highly consistent, straightforward and cost effective cell patterning scheme has been developed. It is based on two common ingredients: the polymer parylene-C and horse serum. Parylene-C is deposited and photo-lithographically patterned on silicon oxide (SiO₂) surfaces. Subsequently, the patterns are activated via immersion in horse serum. Compared to non-activated controls, cells on the treated samples exhibited a significantly higher conformity to underlying parylene stripes. The immersion time of the patterns was reduced from 24 to 3 h without compromising the technique. X-ray photoelectron spectroscopy (XPS) analysis of parylene and SiO₂ surfaces before and after immersion in horse serum and gel based eluant analysis suggests that the quantity and conformation of proteins on the parylene and SiO₂ substrates might be responsible for inducing glial and neuronal patterning.     

蒲黄总黄酮对3T3-L1脂肪细胞糖脂代谢的影响

目的探讨蒲黄总黄酮对脂肪细胞糖脂代谢的影响。方法药物处理胰岛素抵抗3T3-L1脂肪细胞24h,通过检测培养液中的葡萄糖含量反映葡萄糖消耗量;XTT法观察药物对细胞活性的影响;3H-葡萄糖摄入法观察细胞对葡萄糖的转运率;通过检测上清液中的浓度观察细胞游离脂肪酸(freefattyacid,FFA)的溢出。结果蒲黄总黄酮在浓度为0.025~0.4g/L时有增加葡萄糖消耗的作用,且呈剂量效应。当浓度为0.4g/L时,XTT值明显下降,显示对细胞的毒性作用。在0.2g/L时,蒲黄总黄酮与罗格列酮相似,可明显提高细胞对3H-脱氧葡萄糖的转运率。蒲黄总黄酮组培养上清液中FFA浓度明显低于空白对照组,而罗格列酮组则无明显差异。结论蒲黄总黄酮能显著增加3T3-L1脂肪细胞的葡萄糖摄取和消耗,同时可减少FFA溢出,通过调节糖代谢和脂代谢改善胰岛素抵抗。     

电针对关节炎大鼠脊髓背角环氧合酶-2表达的影响

目的 本工作试图通过观察慢性炎症痛大鼠脊髓背角环氧合酶（COX）-2蛋白表达的变化，探讨脊髓背角COX-2是否与电针镇痛的作用机制有关。方法 在佐剂性关节炎大鼠的模型上，采用行为学和免疫组化技术两种方法，观察电针治疗后不同时相点关节炎大鼠的痛敏评分及脊髓背角COX-2蛋白的表达，并与正常组、模型组和药物组加以对照。结果 电针能明显减小关节炎大鼠痛敏评分的绝对值，多次电针具有显著的累积效应；并且随着电针治疗次数的增加，大鼠脊髓背角COX-2免疫阳性细胞数显著下降。结论 电针阳陵泉和昆仑两穴对佐剂性关节炎大鼠有明显的镇痛作用，并同时抑制其脊髓背角COX-2蛋白的表达。     

重视同病类证和同病类治的研究

病证关系除“同病异证”和“异病同证”外，还表现为“同病类证”。同病类证是指同一疾病患者具有相同的中医主证，个体之间只是兼夹证的差异，因此治疗上可运用中医类治法，即在应用主方治疗的基础上根据兼夹证适当加减。同病类证强调病和证的同质性，使辨病治疗和辨证治疗有更高的融合度。加强对同病类证和同病类治的研究是中西医结合的一项重要内容，有利于用现代科学方法阐明中医证的内涵，也有利于发扬中医辨证施治的优势和个体化治疗的特色。同病类证和同病类治对临床医师掌握方证相应的辨证能力提出了更高的要求。     

Associations between cytokines,endocrine stress response,and gastrointestinal symptoms in autism spectrum disorder

Many children and adolescents with autism spectrum disorder (ASD) have significant gastrointestinal (GI) symptoms, but the etiology is currently unknown. Some individuals with ASD show altered reactivity to stress and altered immune markers relative to typically-developing individuals, particularly stress-responsive cytokines including tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6). Acute and chronic stress is associated with the onset and exacerbation of GI symptoms in those without ASD. The present study examined whether GI symptoms in ASD were associated with increases in cortisol, a stress-associated endocrine marker, and TNF-α and IL-6 in response to stress. As hypothesized, a greater amount of lower GI tract symptoms were significantly associated with post-stress cortisol concentration. The relationship between cortisol response to stress and GI functioning was greater for children who had a history of regressive autism. Exploratory analyses revealed significant correlations between cortisol response, intelligence, and inappropriate speech. In contrast, symptoms of the lower GI tract were not associated with levels of TNF-α or IL-6. Significant correlations were found, however, between TNF-α and IL-6 and irritability, socialization, and intelligence. These findings suggest that individuals with ASD and symptoms of the lower GI tract may have an increased response to stress, but this effect is not associated with concomitant changes in TNF-α and IL-6. The relationship between cortisol stress response and lower GI tract symptoms in children with regressive autism, as well as the relationships between cortisol, IL-6, and intelligence in ASD, warrant further investigation.     

Primary murine microglia are resistant to nitric oxide inhibition of indoleamine 2,3-dioxygenase

Indoleamine 2,3-dioxygenase (IDO) is an intracellular heme-containing enzyme that is activated by proinflammatory cytokines, including interferon-γ (IFNγ), and metabolizes tryptophan along the kynurenine pathway. Activation of murine macrophages induces not only IDO but also nitric oxide synthase (iNOS), and the ensuing production of nitric oxide (NO) inhibits IDO. To determine the sensitivity of primary cultures of murine microglia to NO, microglia were stimulated with recombinant murine IFNγ (1 ng/ml) and lipopolysaccharide (LPS) (10 ng/ml). This combination of IFNγ + LPS synergized to produce maximal amounts of nitrite as early as 16 h. Steady-state mRNAs for both iNOS and IDO were significantly increased by IFNγ + LPS at 4 h post-treatment, followed by an increase in IDO enzymatic activity at 24 h. Murine microglia (>95% CD11b⁺) were pretreated with the iNOS inhibitor, L-NIL hydrochloride, at a dose (30 μM) that completely abrogated production of nitrite. L-NIL had no effect on IDO mRNA at 4 h or IDO enzymatic activity at 24 h following stimulation with IFNγ + LPS. These data establish that IDO regulation in murine microglia is not restrained by NO, thereby permitting the accumulation of kynurenine and its downstream metabolites in the central nervous system.