5-氟尿嘧啶诱导结肠癌细胞系SW 620凋亡的傅里叶变换红外光谱研究

探索红外光谱监测肿瘤细胞凋亡的可行性及初步分析。采用结肠癌细胞系SW620作为诱导细胞模型,MTT法确定最优5-FU诱导浓度, 无血清饥饿培养协同细胞周期阻滞在G1和S期,傅里叶显微红外光谱仪(FTIR) 联合流式细胞仪(FCM)对SW 620细胞和凋亡的SW 620细胞12 h、早期凋亡(24 h)和晚期凋亡(48 h)的峰位、相对峰强等指标进行比较分析。光谱分析结果显示SW 620对比凋亡细胞有以下特征：(1)与脂类相关谱带1 740 cm-1相对峰强比I_{1 740}/I_{1 460}明显降低(p<0.05),表明凋亡细胞中脂类相对含量增多。(2)与氨基酸残基相关谱带1 410 cm-1,I_{1 460}/I_{1 460}在凋亡早期和晚期明显升高(p<0.05), 与丝、苏氨基酸相关谱带1 120 cm-1 向高波数移动,I_{1 120}/I_{1 460}明显升高(p<0.05),表明凋亡细胞中DNA双螺旋解链,氨基酸残基增多。(3)DNA的反对称伸缩1 240 cm-1向高波数移动,表明凋亡细胞中核酸分子构象发生改变。(4)与核酸多糖1 040 cm-1相关谱带在凋亡的24和48 h出现,向高波数移动,I_{1 040}/I_{1 460}在凋亡晚期降低(p<0.05)。初步研究结果表明傅里叶变换红外光谱分析可能成为实时无创监测肿瘤化疗的有效方法。     

红外光谱用于大肠肿瘤诊断的初步研究

探索红外光谱诊断大肠肿瘤性质的可行性。联合使用衰减全反射探头与傅里叶变换红外光谱仪测定新鲜离体大肠18个癌组织和10个良性腺瘤的红外光谱,将两组光谱的峰位及相对峰强等指标进行比较。与良性腺瘤相比,大肠癌的红外光谱具有如下特征:(1)与脂类相关的谱带2 925和1 740cm-1峰强比I2 925/I1 460(p=0.018)和I1 740/I1 460(p=0.009)明显降低,表明癌组织的脂类相对含量降低;(2)与蛋白质相关峰强比值I3 375/I1 460(p=0.012)和I1 550/I1 460(p=0.041)明显升高,表明蛋白质相对含量明显升高;(3)与核酸相关的1 080cm-1处峰位发生明显的蓝移(p=0.039),峰强比I1 080/I1 460明显升高(p=0.036),说明癌组织中核酸相对脂类含量明显增加;(4)谱带1 305cm-1处峰位发生明显的红移(p=0.041),其指认有待进一步研究。初步研究结果表明,傅里叶变换红外光谱分析可能成为一种快速鉴别大肠肿瘤性质的新方法。     

胆囊癌的傅里叶变换红外光谱研究

运用衰减全反射(attenuated total reflection,ATR)探头与傅里叶变换红外(Fourier transform infra-red,FTIR)光谱仪,测定并分析了新鲜离体的胆囊癌组织18例和良性组织139例的FTIR光谱。结果表明:(1)胆囊癌的1 167和1 123 cm-1谱带的峰位显著地向低波数移动(P0.05),而1 309 cm-1谱带的峰位显著地向高波数移动(P0.05)。(2)胆囊癌组织光谱多个峰的相对强度I2 856/I1 461,I1 167/I1 461,I1 123/I1 461和I1 082/I1 461明显升高(P0.05)。(3)1 167和1 082 cm-1谱带的半高宽显著升高(P0.05),谱带1 461 cm-1的半高宽则显著降低(P0.05)。(4)癌组织1 750 cm-1谱带的出现几率明显增加(P0.05)。与胆囊良性组织相比较,癌组织光谱中与脂类、糖和核酸相关的谱带均发生了明显的变化。     

结节性甲状腺肿的傅里叶变换红外光谱体表测定

利用ATR探头和中红外光导纤维红外光谱法在体表测定34个正常甲状腺和20个结节性甲状腺肿的傅里叶变换红外光谱。并比较了二者之间13条谱带的27个指标的差异。结果表明: 结节性甲状腺肿的体表红外光谱中：(1)～2 925 cm-1 谱带(—CH₂的反对称伸缩振动)的峰位, ～1 250 cm-1(PO的伸缩振动)谱带的峰位均明显地向低波数方向移动(P＜0.05);(2)谱带的相对强度(谱峰高度)比值H_{1 740}/H_{1 460}, H_{1 160}/Ｈ_{1 460}和H_{1 160}/H_{1 120} 较正常甲状腺明显降低(P＜0.05);(3)H_{1 080}/H_{1 460}比值明显升高(P=0.008)。这些差异是利用体表红外光谱诊断结节性甲状腺肿的基础。     

辐射损伤小鼠胸腺组织的FTIR光谱变化与辐射剂量的关系

应用傅里叶变换红外(FTIR)光谱分析辐射损伤小鼠胸腺组织随辐射剂量增加的生化变化过程,为临床评估患者辐射剂量提供新的研究思路。结果表明,随着辐射剂量的增加,辐射损伤小鼠胸腺组织FTIR光谱的主要吸收峰位变化表现在照射2,3,5Gy时,在1 550,1 460,1 400及1 640 cm-1处峰位发生移位(P<0.05),峰高差异表现在：(1)与核酸有关的谱带1 085和1 236 cm-1的峰强比呈下降趋势,(2)与蛋白质有关的谱带1 640与1 550 cm-1的峰强比随剂量增加明显下降,(3)与脂类有关的谱带2 958,2 925,1 460及1 400 cm-1未发生明显的变化。研究结果表明傅里叶变换红外光谱有望成为临床估算受照剂量的新方法。     

傅里叶变换红外光谱用于食管癌及癌旁正常组织结构的相关研究

应用傅里叶变换红外光谱法(FTIR)对43例食管癌及癌旁正常组织进行了分析,结果表明,食管癌组织和癌旁正常组织的红外光谱具有较大的差异:正常组织的红外光谱在1550 cm-1附近的酰胺Ⅱ带吸收峰相对较强,峰型高尖,与酰胺Ⅰ带的相对峰高比(I1647/I1550)在癌组织中比较高;正常组织中酰胺Ⅰ带的吸收峰峰位比癌组织的高1-6 cm-1;正常组织中1453 cm-1处的峰强大于1402 cm-1处,而在癌组织中则相反,1402 cm-1附近的吸收峰峰高大于1453cm-1处;癌组织中1080 cm-1附近的峰位和正常组织相比显得高而强,癌组织的相对峰高比(I1080/I1550)高于正常组织。说明傅里叶变换红外光谱法可以用于对良、恶性组织进行鉴别诊断,有希望发展成为肿瘤快速诊断的一种新方法。     

人胸膜间皮瘤与结核型胸膜炎的傅里叶红外光谱分析（英文）

研究以上皮型胸膜间皮瘤,纤维型胸膜间皮瘤,结核型胸膜炎与正常胸膜组织为材料,通过傅里叶变换红外光谱分析组织中生物大分子的结构及含量的改变。研究发现,四种胸膜组织的傅里叶红外光谱较为相似,但存在明显区别,四种胸膜组织红外光谱数据方差差异极显著(sig.0.001),说明胸膜间皮瘤中生物大分子的含量及结构发生了明显变化,主要包括:(1)胸膜间皮瘤蛋白质酰胺Ⅰ带及酰胺Ⅱ带,核酸1 232cm-1峰强,脂类物质2 922cm-1峰强均显著高于正常人胸膜组织,与正常胸膜组织存在显著区别;纤维型胸膜间皮瘤中,蛋白质酰胺Ⅰ带及酰胺Ⅱ带峰强,与核酸密切相关的1 078cm-1峰强,以及与脂类物质相关的2 922和2 854cm-1峰强均显著高于上皮型胸膜间皮瘤(p0.05);结核型胸膜炎蛋白质酰胺Ⅰ带及酰胺Ⅱ、核酸1 232和1 078cm-1峰强略有增加,但与正常胸膜组织差异不显著(p0.05),与脂类物质含量有关2 922cm-1峰强、2 854cm-1峰强,极显著地高于正常胸膜组织(p0.01),显著高于胸膜间皮瘤(p0.05)。(2)蛋白质、核酸、脂类物质的相对峰强I1 641/I2 922,I1 641/I1 232,I1 232/I1 078,I1 078/I1 546,I1 078/I2 854,I2 922/I1 232,I1 458/I1 400能有效放大四类胸膜组织间的差异,其效果优于峰强效果,可作为胸膜间皮瘤诊断的优化指标。(3)上皮型胸膜间皮瘤中指示核酸分子中磷酸二酯键的C—C/C—O的1 078cm-1峰强以及指示脂类物质的2 854cm-1峰强显著低于纤维型胸膜间皮瘤和正常胸膜组织(p0.05),表明上皮型胸膜间皮瘤中磷酸二酯键断裂程度较高,DNA受损严重,膜脂过氧化降解明显。说明上皮型胸膜间皮瘤恶化程度高于纤维型胸膜间皮瘤。(4)胸膜间皮瘤蛋白质酰胺Ⅰ带、Ⅱ带谱带、核酸1 232cm-1峰、脂类物质1 458cm-1处CH2振动及1 400cm-1处CH3振动红移,说明蛋白质分子间的氢键受到破坏,核酸分子的氢键结合力减弱,核酸分子的双链结构受到一定程度的破坏,肿瘤组织中膜脂的亚甲基链趋向无序。(5)傅里叶红外光谱能有效区分纤维型胸膜间皮瘤、上皮型胸膜间皮瘤、结核型胸膜炎、正常胸膜组织,为胸膜间皮瘤与结核型胸膜炎的早期、快速诊断提供了可靠的数据。     

甲状腺癌的傅里叶变换红外光谱研究

应用傅里叶变换红外光谱仪,测定了17例甲状腺癌和23例良性甲状腺疾病术中新鲜离体组织的红外光谱。通过用统计学方法对比分析,发现甲状腺良恶性组织的傅里叶红外光谱之间存在明显差异,其中恶性肿瘤光谱有以下特征：(1)甲状腺癌组织的酰胺Ⅰ带明显红移(P<0.01),酰胺Ⅱ带却出现蓝移(P<0.05),癌组织光谱中I_{1 640}/I_{1 460}和I_{1 640}/I_{1 550}较良性组织明显升高(P<0.01),说明恶性肿瘤不仅蛋白质结构上发生了变化,蛋白质的量化上也有明显变化;(2)与脂类相关的2 955, 2 920, 2 870, 2 850和1 740 cm-1谱带出现概率明显变低,表明癌组织的脂类相对含量降低;(3)与核酸相关的1 241 cm-1谱带明显蓝移至(1 238.29±2.87)cm-1 ,I_{1 080}/I_{1 460}较良性肿物组织有升高(P<0.05),表明癌组织中磷酸二酯基团中PO₂增加,因为在癌细胞分裂增生加快,细胞核内DNA含量增加。研究结果表明,红外光谱有望成为术中快速诊断甲状腺恶性肿瘤的主要方法。     

甲状腺癌转移性淋巴结的红外光谱研究

联合运用衰减全反射(attenuated total reflectance,ATR)探头与FTIR光谱仪,测定了新鲜离体的甲状腺癌转移性颈部淋巴结20枚和非转移性淋巴结69枚的FTIR光谱,比较两组光谱13个谱带的峰位及相对峰强等28个指标,找出甲状腺癌转移性淋巴结光谱的特征。与非转移性淋巴结相比,转移性淋巴结光谱中与蛋白、脂质、糖类、核酸相关的谱带均发生了明显的变化,具体如下：(1)与蛋白质相关的谱带的相对峰强I_{3 280}/I1 460(P=0.029),I_{1 640}/I_{1 460}(P=0.019),I_{1 546}/I_{1 460}(P=0.004)均显著升高;(2)脂类相关的相对峰强I_{1 743}/I_{1 460}显著降低(P=0.026),而I_{1 400}/I_{14 60}则明显增高(P=0.001);(3)与糖类相关的1 165与1 120 cm-1处峰位P_{1 165}(P=0.019),P_{1 120}(P=0.000)发生明显蓝移(向高波数移动),相对峰强I1 165/I_{1 460}显著降低(P=0.006);(4)与核酸相关的1 085 cm-1处峰位P_{1 085}发生明显地红移(向低波数移动)(P=0.029);(5)其他指认未明的谱带的相对峰强I_{1 303}/I_{1 460}(P=0.008),I_{1 303}/I1 240(P=0.039)均明显升高。与非转移性淋巴结光谱相比较,甲癌转移的淋巴结光谱在多个谱带的峰位或相对峰强方面存在显著的差异。FTIR可应用于术中甲状腺癌转移性淋巴结的探测。     

胆管癌新鲜离体组织的傅里叶变换红外光谱研究

该研究探讨了运用傅里叶变换红外(FTIR)光谱技术对胆管癌进行术中原位、无创和快速诊断的可行性。联合应用傅里叶变换红外光谱仪和衰减全反射(ATR)探头,测定了26例胆管癌和43例良性胆管疾病新鲜离体组织的红外吸收光谱,发现每条光谱在3 800～1 000 cm-1之间循序出现了12个谱带。测量各个谱带的峰位、峰强和峰面积值,然后计算出各谱带的相对峰强和相对峰面积比值,最后进行标准统计学分析。比较胆管癌与良性胆管组织的光谱发现有以下特征：(1)2 925 cm-1谱带明显地向低波数移位(P=0.033);(2)峰强比I_{1 083}/I_{1 460}(P=0.005)和峰面积比S1 083/S_{1 460}(P=0.001),S_{1 240}/S_{1 460}(P=0.025)明显升高,说明癌组织中核酸相对脂类含量明显增加;(3)峰强比I_{1 550}/I_{1 083}(P=0.000)和峰面积比S_{1 550}/S_{1 083}(P=0.000)明显下降,提示蛋白质相对核酸的含量下降。研究结果表明,FTIR有望成为一种术中原位、在体和快速诊断胆管癌的新方法。     

Cell patterning without chemical surface modification: Cell-cell interactions between printed bovine aortic endothelial cells (BAEC) on a homogeneous cell-adherent hydrogel

Cell printing offers the unique ability to directly deposit one or multiple cell types directly onto a surface without the need to chemically pre-treat the surface with lithographic methods. We utilize biological laser printing (BioLP^™) to form patterns of bovine aortic endothelial cells (BAECs) onto a homogeneous cell adherent hydrogel surface. These normal cells are shown to retain near-100% viability post-printing. In order to determine whether BAECs encountered shear and/or heat stress during printing, immunocytochemical staining experiments were performed to detect potential expression of heat shock proteins (HSP) by the deposited cells. Printed BAECs expressed HSP at levels similar to negative control cells, indicating that the BioLP process does not expose cells to damaging levels of stress. However, HSP expression was slightly higher at the highest laser energy studied, suggesting more stress was present under these extreme conditions. Printed BAECs also showed preferential asymmetric growth and migration towards each other and away from the originally printed pattern, demonstrating a retained ability for the cells to communicate post-printing.     

荧光偏振光谱法探测光动力过程中癌细胞膜的流动性

以DPH作荧光探剂,用荧光偏振光谱法研究了金属酞菁配合物苯硫基钛菁锌(C₅₆H₃₂N₈S₄Zn)在光动力过程中,不同光照时间对乳腺癌细胞膜流动性的影响。实验结果表明,经光照激发光敏剂金属酞菁后,荧光标记团的偏振度增大,癌细胞膜流动性降低,微黏度升高。说明这是光动力治疗的疗效所在。用酶联免疫检测(MTT法)不同光照时间对细胞存活的影响,结果显示癌细胞最低存活率与膜流动性降低成正相关。这表明光动力过程在一个相近的程度影响癌细胞膜的流动性和增殖,提示光动力过程可能通过影响细胞膜流动性引起细胞增殖变化,即细胞膜是光动力作用的一个位点。     

发酵过程中菌液浓度的检测

采用分光光度测量发酵菌液浓度,选择波长215nm,首先测定样品中菌液的吸光度,再根据校准曲线获得菌体浓度。该法设备简单,操作方便、快速,测量相对误差小于3%,相对标准偏差小于2%,精密度高,准确度好,适合于生产监控。     

The study of low temperature hydrothermal growth of ZnO nanorods on stents and its applications of cell adhesion and viability

The hydrothermal growths of the ZnO nanorods with the densities ranging from 157 to 73 nanorods/μm² were achieved by diluting the ZnO seed solution. However, the ZnO seed nanocrystals started to agglomerate for the seed solution diluted below 1% of the original nano-crystalline solutions and resulted in the formation of clustered nanorods. With the assistance of a surfactant, Triton X-100, the nanorod density can be further reduced to 4 nanorods/μm². The diameters of the nanorods depended on the concentration of the seed solution and agitation speed of the nanorod growth solution. More diluted seed solution used and less agitation of the growth solution, the larger diameter of the nanorods was obtained. This indicated that the nanorod growth mechanism was controlled by the diffusion of reactants. With sufficient agitation of the growth solution, the nanorod can be uniformly grown with subjects on any arbitrary geometry. We have demonstrated ZnO nanorods growth on both inside and outside of biliary stents as well as on nitinol wires used as metal stents. The effect of nanorod density on the NIH 3T3 and HUVEC cells growth was also investigated in this study and the results suggested nanorod-coating to be a suitable method for controlling cell adhesion and viability on implantable devices.     

ANXA2 enhances the progression of hepatocellular carcinoma via remodeling the cell motility associated structures

Hepatocellular carcinoma (HCC) ranks as the fifth most common malignancy worldwide. The detailed mechanism of signal regulation for HCC progression is still not known, and the high motility of cancer cells is known as a core property for cancer progression maintenance. Annexin A2 (ANXA2), a calcium-dependent phospholipids binding protein is highly expressed in HCC. To study the roles the excessively expressed ANXA2 during the progression of HCC, we inhibited the ANXA2 expression in SMMC-7721 cells using RNAi, followed by the analysis of cell growth, apoptosis and cell motility. To explore the relationship between the cell behaviors and its structures, the microstructure changes were observed under fluorescence microscopy, laser scanning confocal microscopy and electron microscopy. Our findings demonstrated that down-regulation of ANXA2 results in decreased the cell proliferation and motility, enhanced apoptosis, suppressed cell pseudopodia/filopodia, inhibited expression of F-actin and β-tubulin, and inhibited or depolymerized Lamin B. The cell contact inhibition was also analyzed in the paper. Take together, our results indicate that ANXA2 plays an important role to enhance the malignant behaviors of HCC cells, and the enhancement is closely based on its remodeling to cell structures.     

基于紫外吸收光谱的癌细胞模型研究

细胞周期是生命起源最基本也最重要的过程,是细胞全部生理过程的综合体现,文章从紫外吸收光谱随其细胞周期的变化入手,以一种简单且能准确反映癌细胞的生长变化过程的方式建模。通过人工诱导细胞同步化方法对子宫颈癌细胞进行处理,获取了处于G1期、S期、G2期和M期的同步化细胞样品,通过测量这些样品的紫外-可见光光谱,对不同周期细胞紫外-可见光光谱的吸光度随细胞周期中芳香族氨基酸、蛋白质及核酸等物质的变化规律进行了分析。根据细胞周期中不同阶段样品对应紫外-可见光光谱在204和260 nm处的吸光度和时间点的关系,利用分段线性回归法建立了子宫颈癌细胞紫外吸收光谱模型。可对细胞周期进行判断,为对细胞周期进行分析研究提供了新的依据,也为细胞建模提供了新思路。     

Models of collective cell motion for cell populations with different aspect ratio: Diffusion,proliferation and travelling waves

Continuum, partial differential equation models are often used to describe the collective motion of cell populations, with various types of motility represented by the choice of diffusion coefficient, and cell proliferation captured by the source terms. Previously, the choice of diffusion coefficient has been largely arbitrary, with the decision to choose a particular linear or nonlinear form generally based on calibration arguments rather than making any physical connection with the underlying individual-level properties of the cell motility mechanism. In this work we provide a new link between individual-level models, which account for important cell properties such as varying cell shape and volume exclusion, and population-level partial differential equation models. We work in an exclusion process framework, considering aligned, elongated cells that may occupy more than one lattice site, in order to represent populations of agents with different sizes. Three different idealisations of the individual-level mechanism are proposed, and these are connected to three different partial differential equations, each with a different diffusion coefficient; one linear, one nonlinear and degenerate and one nonlinear and nondegenerate. We test the ability of these three models to predict the population-level response of a cell spreading problem for both proliferative and nonproliferative cases. We also explore the potential of our models to predict long time travelling wave invasion rates and extend our results to two-dimensional spreading and invasion. Our results show that each model can accurately predict density data for nonproliferative systems, but that only one does so for proliferative systems. Hence great care must be taken to predict density data with varying cell shape.     

Quantifying cell adhesion through forces generated by acoustic streaming

The strength of cell adhesion is important in understanding the cell’s health and in culturing them. Quantitative measurement of cell adhesion strength is a significant challenge in bioengineering research. For this, the present study describes a system that can measure cell adhesion strength using acoustic streaming induced by Lamb waves. Cells are cultured on an ultrasound transducer using a range of preculture and incubation times with phosphate-buffered saline (PBS) just before the measurement. Acoustic streaming is then induced using several Lamb wave intensities, exposing the cells to shear flows and eventually detaching them. By relying upon a median detachment rate of 50 %, the corresponding detachment force, or force of cell adhesion, was determined to be on the order of several nN, consistent with previous reports. The stronger the induced shear flow, the more cells were detached. Further, we employed a preculture time of 8 to 24 h and a PBS incubation time of 0 to 60 min, producing cell adhesion forces that varied from 1.2 to 13 nN. Hence, the developed system can quantify cell adhesion strength over a wide range, possibly offering a fundamental tool for cell-based bioengineering.     

超声对细胞膜通透性的影响及应用

适当参数的超声，使细胞膜发生可以修复的损伤，使细胞膜的通透性提高，从而使细胞内的物流释放到细胞外，细胞外的物质进入细胞内，这一现象可以用于释放代谢产物或中间产物，基因转导，强化化学方法治疗肿瘤等方面。