基于磷脂酰肌醇 3-激酶（ PI3K）/丝苏氨酸蛋白激酶（ AKT）信号通路探究安罗替尼对 A549细胞增殖、凋亡的影响。将 A549细胞分为对照组、各剂量试验组、安罗替尼组、阳性药物组、抑制剂组和激活剂组。干预 24 h后检测 A549细胞活力、细胞形态、增殖率、凋亡情况及相关蛋白的表达水平。结果显示： 20 μmol/L安罗替尼处理后， A549细胞活力降低；安罗替尼组和阳性药物组较对照组 A549细胞生长受到抑制，细胞增殖率、细胞周期素 D1（Cyclin D1）及 p-PI3K、p-AKT的表达水平降低，细胞凋亡数量和半胱氨酸天冬氨酸蛋白酶 -3（Caspase-3）的表达水平升高。抑制剂增强了上述各指标的变化，而激活剂则具有与抑制剂相反的趋势。该研究结果说明，安罗替尼可显著抑制人肺癌 A549细胞的增殖并促进其凋亡，其作用机制可能与抑制 PI3K/AKT通路的信号转导相关。该研究结果进一步说明了安罗替尼作为抗肺癌药物的潜在价值，为抗肺癌药物的研发提供了新的理论基础。

The automatic and accurate identification of apoptosis facilitates large-scale cell analysis. Most identification approaches using nucleus fluorescence imaging are based on specific morphological parameters. However, these parameters cannot completely describe nuclear morphology, thus limiting the identification accuracy of models. This paper proposes a new feature extraction method to improve the performance of the model for apoptosis identification. The proposed method uses a histogram of oriented gradient (HOG) of high-frequency wavelet coefficients to extract internal and edge texture information. The HOG vectors are classified using support vector machine. The experimental results demonstrate that the proposed feature extraction method well performs apoptosis identification, attaining 95.7% accuracy with low cost in terms of time. We confirmed that our method has potential applications to cell biology research.The automatic and accurate identification of apoptosis facilitates large-scale cell analysis. Most identification approaches using nucleus fluorescence imaging are based on specific morphological parameters. However, these parameters cannot completely describe nuclear morphology, thus limiting the identification accuracy of models. This paper proposes a new feature extraction method to improve the performance of the model for apoptosis identification. The proposed method uses a histogram of oriented gradient (HOG) of high-frequency wavelet coefficients to extract internal and edge texture information. The HOG vectors are classified using support vector machine. The experimental results demonstrate that the proposed feature extraction method well performs apoptosis identification, attaining 95.7% accuracy with low cost in terms of time. We confirmed that our method has potential applications to cell biology research.

Apoptosis is very important for the maintenance of cellular homeostasis and is closely related to the occurrence and treatment of many diseases. Mitochondria in cells play a crucial role in programmed cell death and redox processes. Nicotinamide adenine dinucleotide (NAD(P)H) is the primary producer of energy in mitochondria, changing NAD(P)H can directly reflect the physiological state of mitochondria. Therefore, NAD(P)H can be used to evaluate metabolic response. In this paper, we propose a noninvasive detection method that uses two-photon fluorescence lifetime imaging microscopy (TP-FLIM) to characterize apoptosis by observing the binding kinetics of cellular endogenous NAD(P)H. The result shows that the average fluorescence lifetime of NAD(P)H and the fluorescence lifetime of protein-bound NAD(P)H will be affected by the changing pH, serum content, and oxygen concentration in the cell culture environment, and by the treatment with reagents such as H2O2 and paclitaxel. Taxol (PTX). This noninvasive detection method realized the dynamic detection of cellular endogenous substances and the assessment of apoptosis.

类风湿关节炎(RA)是一类以滑膜增生、滑膜关节损害、活动功能不全为主要特点的慢性全身性自我免疫炎症性病变。目前, 对于RA的治疗包括药物治疗、外科手术治疗和物理疗法。物理疗法中光疗法由于其无侵袭力、无痛等优势, 被用作辅助治疗手段普遍应用于临床。为此, 本研究通过630 nm LED红光探讨了光生物调节法通过促进成纤维样滑膜细胞凋亡, 从而治愈类风湿性关节炎的机理。JC-1染色试验表明, 630 nm LED光照后线粒体膜电位去极化增强, 膜电位降低; 使用CASP3活性检测试剂盒和CASP9活性检测试剂盒进行试验, 证明630?nm LED可以促进MH7A细胞中CASP3和CASP9酶的活性; 实时荧光定量PCR(RT-qPCR)试验显示, 630?nm LED可以促进瞬时受体电位香草素1(TRPV1)的表达; 蛋白质免疫印迹试验(Western blot)证明, 经过630 nm LED红光照射可以促进MH7A细胞TRPV1蛋白的表达; CCK-8试验表明, 630 nm LED可以显著性抑制MH7A细胞活力, 但不能降低TNF-α刺激下TRPV1拮抗剂处理的MH7A细胞活性; TUNEL试验证实, 630 nm LED照射诱导MH7A细胞凋亡, 但不能诱导TNF-α刺激下TRPV1拮抗剂处理的MH7A细胞凋亡。综上所述, 630 nm LED红光在类风湿性关节炎中通过调控TRPV1促进TNF-α诱导的MH7A细胞凋亡, 这对辅助缓解RA滑膜炎症有一定作用。

通过化学气相沉积法制备了g-C3N4@C-TiO2纳米颗粒, 利用X射线衍射(XRD)、荧光光谱(FS)、透射电子显微镜(TEM)、紫外-可见吸收光谱(UV-Vis)、能谱(EDS)等对纳米颗粒进行表征。分别研究了暗室条件及光照条件下g-C3N4@C-TiO2纳米颗粒对HL60细胞的作用效果。采用CCK-8法探究了一系列质量浓度梯度的纳米颗粒处理HL60细胞后的存活率, 并且通过荧光探针标记技术检测细胞内活性氧水平。试验结果表明, 在C-TiO2表面包裹g-C3N4, 可将TiO2可吸收的光波长范围拓展至可见光波段。制备的g-C3N4@C-TiO2粒径在10～20 nm, 满足其进入细胞的尺寸要求。暗毒性试验表明g-C3N4@C-TiO2纳米颗粒在暗室条件下对细胞的毒性较小, 证明了其具有良好的生物相容性。同时, 与TiO2和C-TiO2纳米颗粒处理组相比, g-C3N4@C-TiO2处理组的光动力灭活效率高达(76.5±1.9)%, 表明其可能成为治疗白血病的潜在光敏剂。

本试验以酿酒酵母(Saccharomyces cerevisiae)为材料, 研究玫瑰花提取物(RE)对砷胁迫下酵母细胞生长和凋亡的影响。试验以抗坏血酸(Vc)溶液为标准抗氧化对照物, 1.0 mmol/L亚砷酸钠(As)为酵母胁迫浓度, 分别设置了酵母对照组、酵母+RE(1.5 g/L)培养组、酵母+Vc(0.1 g/L)培养组、酵母+As(1.0 mmol/L)培养组、酵母+As+RE培养组和酵母+As+Vc培养组。酵母细胞培养24 h后, 利用噻唑兰(MTT)细胞增殖法、平板点样法、荧光素二乙酸/碘化丙锭(FDA/PI)染色法、4', 6-二脒基-2-苯基吲哚(DAPI)细胞核染色法和Annexin V-FITC/PI细胞凋亡流式检测法测定了酵母细胞的生长增殖与凋亡。试验结果显示, RE有效地缓解了砷对酵母细胞的损伤, 极显著提高了砷胁迫下酵母细胞的生长率和存活率(P<0.01), 极显著抑制了砷胁迫下酵母细胞的凋亡(P<0.01)。结果证实, RE对砷胁迫酵母细胞的凋亡有显著抑制作用, 在50%的1,1-二苯基-2-三硝基苯肼(DPPH)清除率下, 其对砷胁迫下酵母细胞的凋亡抑制率显著高于Vc(P<0.05)。因此, RE作为天然植物提取物, 因其绿色、无毒的作用机制, 将在对抗重金属污染、提高机体抗氧化力、缓解氧化损伤等方面有着更广阔的应用前景。

根据细胞的生物吞噬特性,通过研究人肝癌细胞(HepG2)内吞Si纳米粒子的拉曼成像特性,提出了一种新的细胞凋亡检测方法。首先,在Si纳米粒子与细胞共培养过程中,对HepG2细胞施加不同浓度的凋亡诱导剂——黄连素,然后对细胞内吞的Si纳米粒子在520 cm-1处的特征拉曼峰检测成像,通过分析细胞区域拉曼像图像素的平均信号强度,表征细胞内吞Si纳米粒子的能力,再对照流式细胞术的检测结果从而获得在不同浓度细胞凋亡诱导剂作用下的细胞凋亡率。结果表明,随着黄连素浓度的增加,细胞区域拉曼像图的平均信号强度逐渐减弱,说明在HepG2细胞凋亡过程中线粒体的活性不断减弱,能量代谢受阻,造成细胞对Si纳米粒子的内吞能力减弱。特别地,在高浓度(150 μM)黄连素作用下,细胞区域内520 cm-1的特征拉曼峰十分微弱,说明此时HepG2细胞基本丧失内吞Si纳米粒子的能力。因此,基于细胞内吞Si纳米粒子拉曼成像方法,可以对不同环境下的体外细胞进行凋亡检测,将在细胞毒理研究和药效评价方面具有潜在应用。