Carbon blacks as EPR sensors for localized measurements of tissue oxygenation.

New electron paramagnetic resonance (EPR) oximetry probes were identified in the class of carbon black materials. These compounds exhibit very high oxygen sensitivity and favorable EPR characteristics for biological applications. At low pO(2), the linewidth is particularly sensitive to changes in oxygen tension (sensitivity of 750 mG/mmHg). The application of the probes for oximetry was demonstrated in vivo: the pO(2) was measured in muscle in which the blood flow was temporarily restricted as well as in tumor-bearing mice during a carbogen breathing challenge. The responsiveness to pO(2) was stable in muscle for at least 3 months. No toxicity was observed using these materials in cellular experiments and in histological studies performed 2, 7, and 28 days after implantation. In view of their EPR characteristics (high sensitivity) as well as the well-characterized production procedure that make them available on a large scale, these probes can be considered as very promising tools for future developments in EPR oximetry.     

Highly resolved in vivo 1H NMR spectroscopy of the mouse brain at 9.4 T.

An efficient shim system and an optimized localization sequence were used to measure in vivo 1H NMR spectra from cerebral cortex, hippocampus, striatum, and cerebellum of C57BL/6 mice at 9.4 T. The combination of automatic first- and second-order shimming (FASTMAP) with strong custom-designed second-order shim coils (shim strength up to 0.04 mT/cm2) was crucial to achieve high spectral resolution (water line width of 11-14 Hz). Requirements for second-order shim strengths to compensate field inhomogeneities in the mouse brain at 9.4 T were assessed. The achieved spectral quality (resolution, S/N, water suppression, localization performance) allowed reliable quantification of 16 brain metabolites (LCModel analysis) from 5-10-microL brain volumes. Significant regional differences (up to 2-fold, P < 0.05) were found for all quantified metabolites but Asp, Glc, and Gln. In contrast, 1H NMR spectra measured from the striatum of C57BL/6, CBA, and CBA/BL6 mice revealed only small (<13%, P < 0.05) interstrain differences in Gln, Glu, Ins, Lac, NAAG, and PE. It is concluded that 1H NMR spectroscopy at 9.4 T can provide precise biochemical information from distinct regions of the mouse brain noninvasively that can be used for monitoring of disease progression and treatment as well as phenotyping in transgenic mice models.     

c-myc mRNA在成釉细胞瘤中的表达

目的:研究成釉细胞瘤(AB)和牙源性角化囊肿(OKC)中c-mycmRNA的表达,探讨c-myc在AB和OKC中的发生、发展及其生物学意义。方法:使用原位杂交法检测54例AB、16例OKC和7例口腔正常黏膜(NOM)组织中c-mycmRNA的表达,并将AB按原发、复发、恶变分组,结果使用χ2检验进行统计分析。结果:AB、OKC及NOM组织中c-mycmRNA的阳性表达率分别为81.5%(44/54)、75.0%(12/16)和14.3%(1/7),3组比较有显著性差异(χ2=15.488,P<0.05)。原发组AB中c-mycmRNA的阳性表达率为71.0%,复发组为94.7%,恶变组为100.0%,伴随原发、复发、恶变,差异有显著性(χ2=16.912,P﹤0.05)。结论:c-myc表达在AB的发生、发展中有重要作用;c-mycmRNA的表达与AB的临床生物学行为有关,伴随其生物学行为变化,c-mycmRNA表达增强;提示c-myc有可能成为评价预后的有效指标。     

LRIG1下调原代星形细胞瘤细胞增殖的机制

目的观察LRIGl蛋白表达对表皮生长因子（EGF）促肿瘤细胞增殖作用的影响，探讨LRIGl抑制肿瘤细胞增殖的机制。方法原代培养19例星形细胞瘤细胞，用原位杂交检测LRIGl的表达，用噻唑蓝（MTT）法观察EGF对培养细胞的促增殖作用，并分析培养细胞LRIGl表达和EGF干预后的细胞生长率的关系。结果（75．3±11．6）％的培养细胞表达LRIGl蛋白；EGF促进培养细胞增殖（38．0±14．8）％，EGF促细胞增殖的细胞生长率与LRIGl表达程度呈负相关。结论LRIGl蛋白可能通过抑制EGF—EGFR信号抑制肿瘤细胞的增殖。     

活体顺序性荧光标记技术在实验性骨折愈合研究中的应用

目的介绍实验性骨折愈合研究中活体顺序性荧光标记的技术与方法。方法以成年兔胫骨骨折外固定为模型，术后分别以Tetracydin(TC，黄色，25mg／kg)、Calcein(CG，绿色，10mg／kg)、Xylenal(XO，桔色，90mg／kg)、Alizarin—complen(AE，红色，30mg／kg)四种荧光染料于兔颈部皮下定期注射；并于术后第3、6、12及24周活杀动物，解剖出胫骨，梯度乙醇脱水、脱脂，甲基丙烯酸甲脂包埋，切片厚度40～100μm，打磨，荧光显微镜下观察、摄像；同时通过组织学观察、X线摄片了解骨愈合速度与质量。结果顺序性荧光标记显示，加压组中钙化的内、外骨痴均于术后第2周出现于截骨处的远、近断端，但尚未形成骨性连接。亦无骨皮质的改建。外骨痴增生停止于术后12周，且骨皮质的改建至12周已渐减少．至术后24周已基本完成；对照组：内骨痴于术后第3周出现于截骨处远、近断端，无钙化的外骨痴形成，亦无骨皮质的改建；外骨痂增生停止于术后12周，骨皮质改建至24周尚未完成，其结果与组织学观察、X线摄片具有一致性。结论活体顺序性荧光标记技术可以从细胞水平判断新骨的开始时间、骨生长速度及生长方向，并能节约实验动物数量，是实验性骨折愈合研究中较为理想的研究手段。     

肿瘤转移抑制基因KAI1在喉鳞状细胞癌中表达的研究

目的　探讨肿瘤转移抑制基因KAI1在喉鳞状细胞癌 (简称鳞癌 )中的表达及其与之发生、发展的关系。方法　采用原位杂交方法检测 84例原发性喉鳞癌 (primarylaryngealsquamouscellcarcinoma ,PLSCC)、2 7例喉癌前病变不典型增生 (laryngealprecancerouslesion ,LPL)、10例声带息肉(vocalcordpolyp ,VCP)和 10例正常喉黏膜 (normallaryngealtissues ,NLT)石蜡标本组织细胞中KAI1mRNA的表达。结果　NLT、VCP、LPL和PLSCC 4种组织中KAI1阳性表达的积分吸光度值 ( x±s)分别为 (136 2 0 6 8± 36 6 75 5 )、(1336 74 5± 4 2 85 8 5 )、(90 36 8 8± 2 5 70 1 9)和 (6 7880 6± 2 8189 5 ) ,其中NLT组和VCP组之间差异无显著性 (t=0 14 2 ,P >0 0 5 ) ,NLT组和LPL组之间差异有显著性 (t =4 2 81,P <0 0 1) ;PLSCC组中KAI1表达普遍下调 ,且病理分化G1 2组阳性表达水平高于G3组 ;T1 2病变组高于T3 4组 ;颈淋巴结NO组高于N1及N1以上组 ;临床Ⅰ Ⅱ期组高于临床Ⅲ Ⅳ期组 (P值均 <0 0 1)。KAI1表达与患者性别无关 (P >0 0 5 )。结论　KAI1低表达在喉鳞癌的发生、发展中可能起着重要作用 ,可望作为喉鳞癌早期诊断、评估肿瘤细胞侵袭转移潜能及患者病程发展阶段的指标之一。     

Mid-infrared spectroscopy on skin using a silver halide fibre probe in vivo

Background/aim: Mid-infrared spectroscopy is a versatile method for in vivo investigation of skin after topical treatment with skin care products.
Methods: FTIR-spectrometer (Bruker Optics) with a flexible silver halide fibre probe (Infrared Fiber Sensors).
Results: Absorbance spectra from 700 to 3000 cm⁻¹ have been recorded to gain information about proteins (amide-I and amide-II vibrations at 1650 and 1550 cm⁻¹), esters (1740 cm⁻¹), carboxylic acid (1710 cm⁻¹), polyalcohols (1050 cm⁻¹) and hydrocarbons (CH _n vibrations at 2800–3000 cm⁻¹).
Conclusions: Using the particular light guide, we were able to measure for the first time the effects of lip care products on lips directly. Furthermore, water binding and glycerol content of the skin could be determined simultaneously, as well as the replenishment of lipids by lipid-enriched bath oil.     

应用激光显微分离技术在骨肉瘤组织原位筛选噬菌体肽库

目的将激光显微分离(laser capture microdissection, LCM) 技术应用于噬菌体表面呈现肽库的筛选过程,建立一种可以直接在天然组织中筛选肽库的方法。方法将新鲜人骨肉瘤组织块在噬菌体肽库溶液振荡孵育后制成组织冰冻切片,免疫组化染色检测噬菌体在组织中的浸润扩散。改进常规LCM切片处理方法,以冻干法替代酒精/二甲苯法脱水,以期在LCM操作过程中提高切片上噬菌体的存活率。LCM法分离摄取骨肉瘤切片上的肿瘤靶细胞,转染回收这些细胞上特异结合的噬菌体。滴定法检测所筛选的噬菌体对特异性细胞的亲和力。结果利用LCM技术,可以由肿瘤冰冻切片上收集到足够的与特异性噬菌体短肽,应用于噬菌体表面呈现肽库筛选。经过3轮筛选后所获得的噬菌体与人骨肉瘤组织的特异性亲和力提高16倍。结论本研究首次将LCM技术应用于噬菌体表面呈现肽库的筛选,可以使我们直接在新鲜人肿瘤组织中筛选与特定细胞群甚至单个细胞亲合的短肽;同时又避免了天然组织中其他杂质细胞的污染,为研制细胞特异性导向载体提供了一个新的途径。     

Fertility in patients with multiple sclerosis: current knowledge and future perspectives

Abstract The issue of fertility in patients with multiple sclerosis (MS) has not been exhaustively studied. Epidemiological data have suggested that spontaneous fecundity might be reduced; several endocrine and sexual disturbances potentially interfering with reproduction have been evidenced in MS patients of both sexes. Moreover, some medical treatments used in MS (e. g., mitoxantrone, cyclophosphamide) may exert detrimental effects on spermatozoa as well as on oocytes, leading to early impairment of fertility. This review illustrates the factors potentially interfering with fertility in MS and discusses the therapeutic tools that may be used to promote fertility in these patients. The safety of hormonal therapies in MS is also examined. The current applications of assisted reproductive technology (ART) are discussed, including in vitro fertilisation (IVF) techniques. Currently available methods to preserve fertility in patients that undergo cytotoxic treatments by means of sperm/oocyte cryostorage or by ovarian fragment cryopreservation and autografting are considered.