丫啶橙-碘化丙啶双染色法观察大鼠耳蜗基底膜毛细胞活性

目的探讨大鼠耳蜗基底膜毛细胞活性鉴别的实验观察方法。方法采用丫啶橙-碘化丙啶(acri-dine orange-propidiumiodide,AO/PI)双染色法,荧光显微镜观察,检测经组织培养后新生大鼠耳蜗基底膜毛细胞的活性。结果在激发光下染色后的活细胞核是亮绿色,失活细胞核呈橙红色。当染液pH为6.5,AO浓度为0.1‰时,细胞染色不足,浓度超过1.5‰时,出现“过染”,最适染色浓度为1‰;当1‰AO的pH值低于5.0时毛细胞蒙上红晕,背景模糊,当pH值高于7.4时,不能区分死亡细胞与活细胞,pH值6.5~7.0时,染色效果最好。结论用pH为6.8的1‰AO、0.1‰PI染色,可清晰分辨死/活细胞。     

豚鼠Ⅱ型前庭毛细胞乙酰胆碱敏感性钾电流的钙离子依赖性

目的研究豚鼠Ⅱ型前庭毛细胞乙酰胆碱(acetylcholine,ACh)敏感性钾电流对钙离子的依赖性。方法应用全细胞膜片钳技术研究新鲜单离的豚鼠Ⅱ型前庭毛细胞ACh-敏感性钾电流在细胞内外钙离子浓度改变时其电流幅值的变化。结果①细胞外ACh激活一缓慢持久的外向性钾电流,其反转电位为(-70±10)mV;②ACh-敏感性钾电流电流对细胞外四乙胺(10mmol/L)敏感,而对细胞外4-氨基吡啶(100μmol/L)不敏感;③ACh-敏感性钾电流的幅值大小依赖于细胞外的钙离子浓度,无钙外液中ACh激活一很小的电流,4mmol/L外钙溶液中ACh-敏感性钾电流的幅值达到最大值,而0·5mmol/L外钙溶液中ACh-敏感性钾电流的幅值抑制至(36·5±6·5)%;④细胞内三磷酸肌醇-钙离子释放过程不参与ACh-敏感性钾电流的激活,细胞内透析肝素8mg/ml后30min,ACh-敏感性钾电流的幅值没有明显改变;⑤ACh敏感性钾电流对钙通道阻断剂Cd2+敏感。结论豚鼠Ⅱ型前庭毛细胞ACh-敏感性钾电流的激活依赖于细胞外的钙离子浓度,ACh与豚鼠Ⅱ型前庭毛细胞胆碱能受体结合后,首先激活膜上钙通道引起细胞外钙离子内流,毛细胞内游离钙离子浓度的升高进一步激活钙依赖性钾电流。     

婴幼儿头发矿物质及微量元素含量测定分析

目的了解婴幼儿体内矿物质及微量元素的含量,为合理安排膳食营养和科学喂养提供依据。方法采用原子吸收分光光度法对138名婴幼儿头发进行检测。结果钙、锌、镁、铁等元素缺乏。结论不良饮食和家长缺乏营养知识是影响婴幼儿矿物质及微量元素缺乏的主要原因。     

红花和当归等中药煎剂对体外培养的鼠毛囊毛发生长与毛球部细胞增殖的影响

目的:探讨红花、当归等中药混合煎剂和单味煎剂,对体外培养的小鼠毛囊毛发生长与毛囊球部细胞生长活力的影响。方法:将离体培养的小鼠触须毛囊和乳鼠背部皮肤的毛囊球部细胞,分别设为对照组(W illiam s培养液)和中药组(W illiam s培养液+不同的中药煎剂)即混合组1(1.5×1-0 5g/m l),混合组2(1.5×1-0 6g/m l),混合组3(1.5×1-0 7g/m l),当归组与红花组分别为1.5×1-0 4g/m l、1.5×1-0 5g/m l,生侧柏叶组为5.0×1-0 4g/m l、5.0×1-0 5g/m l。显微镜下观察各组毛囊毛发生长情况,MTT比色试验测定各组吸光度A值。结果:毛囊培养第7天,中药混合组的毛囊毛发生长长度大于对照组和单味煎剂组(P<0.05),1.5×1-0 5g/m l混合组的毛囊毛发生长长度大于1.5×1-0 7g/m l组(P<0.05);单味中药组与对照组之间的毛囊毛发生长长度相近。毛囊球部细胞培养72 h、96 h中药混合组的吸光度A值高于对照组(P<0.05),72 h混合组1.5×1-0 5g/m l与1.5×1-0 7g/m l的吸光度A值高于部分单药组(P<0.05);单味中药组与对照组之间以及3个浓度中药混合组之间的吸光度A值相似。结论:红花、当归等中药混合煎剂对体外培养的小鼠毛囊毛发生长起促进作用,可促进毛囊球部细胞增殖;单味中药煎剂对体外培养的小鼠毛囊毛发生长及毛囊球部细胞增殖无明显作用。     

Determination of aromatic amines in hair dye and henna samples by ion-pair extraction and gas chromatography-mass spectrometry

A gas chromatography-mass spectrometry (GC-MS) method has been proposed for the determination of carcinogenic and toxic aromatic amines in hair dye, henna and dyed hair samples. The method includes ion-pair extraction of aromatic amines from aqueous samples with bis-2-ethylhexylphosphate (BEHPA) released after solving the samples in acidic solution followed by sonication, derivatisation of compounds with isobutyl chloroformate (IBCF) and their GC-MS analysis in both electron impact (EI) and positive and negative ion chemical ionisation (PNICI) mode as their isobutyloxycarbonyl (isoBOC) derivatives. The obtained recoveries of aromatic amines ranged from 92.2 to 98.4% and the precision of this method, as indicated by the relative standard deviations (RSDs) was within the range of 0.7-4.2%. The detection limits obtained from calculations by using GC-MS results based on signal-to-noise ratio (S/N)=3 were within the range from 0.02 to 0.20 ng/g. In the present study, the commercially available 54 permanent hair dye, 35 modified or natural henna and 15 dyed hair samples were analysed for the aromatic amines by the proposed method and the method was shown to be suitable to determine the aromatic amine ingredients and metabolites of these commercial products.     

Effect of long-term geomagnetic field deprivation on the concentration of some elements in the hair of laboratory rats

The aim of the study was to determine the effect of long-term geomagnetic field (GMF) deprivation on the concentration of selected elements in the hair of laboratory rats. A total of 32 Wistar laboratory rats were divided into four equal groups (males and females) kept under hypomagnetic conditions (GMF vertical component below 20 nT) and two control groups (males and females) kept free of field disturbances (GMF vertical component approx. 38 000 nT). At the beginning and at 7 months of the experiment, hair was taken from the dorsal part of all rats and analysed using atomic emission spectrometry for the concentration of selected magnetic elements (Fe, Ni, Co, Cr, Mn and Cu). Long-term GMF deprivation was found to affect the concentration of Fe, Mn, Cu and Cr, but had no significant effect on the concentration of Co or Ni in the hair of the analysed rats.     

猪脱细胞真皮基质对小鼠创面毛囊再生中基质细胞衍生因子-1及Wnt3a/β-catenin信号通路表达的影响

目的探讨猪脱细胞真皮基质(ADM)对小鼠创面毛囊再生中基质细胞衍生因子-1(SDF-1)及Wnt3a/β-catenin信号通路表达的影响。 方法取新鲜猪皮,经脱细胞处理后制成微粒状,灭菌,密封,常温保存,以备后面实验使用。制作18只C57BL/6小鼠背部全层皮肤缺损模型,以脊柱为中线,在左右各制作直径为6 mm的缺损,左右侧分别以纱布和猪ADM覆盖,隔天纱布侧换药,猪ADM侧不做处理,于模型建立第7天,按照所用材料不同分为纱布组与猪ADM组,再按所取部位不同分为纱布窗口组、猪ADM窗口组、纱布创面组、猪ADM创面组、纱布创缘组、猪ADM创缘组,其中9只小鼠组织用于蛋白质印迹法检测,另外9只小鼠组织用于免疫组织化学检测。通过蛋白质印迹法检测肿瘤坏死因子-α(TNF-α)、β-catenin、转化生长因子-β(TGF-β)、Wnt3a、血管内皮生长因子(VEGF)、SDF-1、成纤维细胞生长因子(FGF)2、FGF9、AKT的蛋白表达,免疫组织化学检测Wnt3a、SDF-1、FGF2、FGF9的表达。数据比较采用独立样本t检验。 结果在小鼠模型建立第7天创面中,蛋白质印迹法检测β-catenin、Wnt3a、SDF-1的蛋白表达量均是猪ADM窗口组(0.533±0.058、0.446±0.039、0.972±0.048)高于纱布窗口组(0.401±0.005、0.132±0.022、0.175±0.036),差异均有统计学意义(t＝3.996、12.230、23.130,P值均小于0.05)。β-catenin、Wnt3a、SDF-1的蛋白表达量均是猪ADM创面组(0.557±0.009、0.626±0.066、0.868±0.102)高于纱布创面组(0.302±0.010、0.109±0.019、0.036±0.009),差异均有统计学意义(t＝32.830、13.020、14.130,P值均小于0.05)。Wnt3a、SDF-1的蛋白表达量均是猪ADM创缘组(0.419±0.014、0.370±0.069)高于纱布创缘组(0.115±0.020、0.056±0.007),差异均有统计学意义(t＝21.460、7.825,P值均小于0.05)。免疫组织化学检测结果显示Wnt3a、SDF-1、FGF2、FGF9在猪ADM组中的表达高于纱布组,且阳性细胞主要分布于毛囊细胞周围。 结论猪ADM在小鼠创面中可能通过上调SDF-1及Wnt3a/β-catenin信号通路的表达而促进毛囊的再生。     

Expression of bFGF and NGF and their receptors in chick's auditory organ following overexposure to noise

Growth factors are known to activate signaling cascades for DNA replication; they participate in the regulation of cell differentiation and are required as positive signals for cell survival. Thus, many of them may be regarded as potential candidates stimulating regeneration processes in the inner ear. We analyzed the expression of basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) and their receptor (bFGFR and NGFR)-like immunoreactivity in chick basilar papillae, along with bFGF and NGF mRNA expression. The evaluation was made 1 and 5 days after exposure to wide-band noise with two increasing levels of acoustic energy.

For both factors, the immunoreactivity was shown predominantly in the middle part of basilar papilla, in noise-exposed, but not control birds. It was localized in the cytoplasm of hair cells, nuclei of supporting cells and cytoplasm of ganglion cells. Strong immunoreactivity of bFGFR and NGFR was found both in control and noise-exposed animals, with the cell localization similar to that of growth factors. The increase in mRNA expression for bFGF and NGF was found in noise-exposed animals only after lower exposure to noise, on day 5 after exposure (p < 0.01). A lack of increased expression after higher exposure could be excused by larger damage of hair cells followed by the increase of mRNA for β-actin to which the results were referred.

The results suggest bFGF and NGF involvement in postinjury regeneration of the basilar papilla.     

A place for fiber diffraction in the detection of breast cancer?

AIMS: To review the potential relevance and the place that a particular change in the fiber diffraction of hair might have in the future management of breast cancer and other pathologies. METHOD: A comprehensive overview was obtained using a complete search of the Australian National University Library (data range) and Medline (data range) using the search terms "review breast cancer screening/diagnosis/detection" and "X-ray diffraction of hair". Publications in the past 5 years were selected but older reports that were commonly referenced were not excluded. RESULTS: To date, our results have demonstrated that a specific change occurs in the diffraction pattern of hair for persons with breast cancer. Further research has shown that this change is present in the hair at an earlier stage of the cancer growth than is detectable by mammography. In addition, the change has been found to disappear when the cancer has been successfully removed. DISCUSSION/CONCLUSION: This technology uses only a few hairs, is totally user-friendly, non-invasive and does not require the patient to be exposed to any radiation. As a low-risk procedure, it could potentially provide a much needed, cost-effective early screening test for the presence of breast cancer in women of any age. As the patients are not required to be present during the test, it could also provide a testing service for women in remote areas. In this review, the origin of the diffraction pattern and the diagnostic information that can be gleaned from it are outlined.