利用AFM和SNOM对淋巴细胞膜表面超微结构及其光学性质的初步研究

利用原子力显微镜(Atomic Force Microscopy，AFM)对淋巴细胞表面形貌进行了形态学的初步研究，观察到了其膜表面其他显微技术所不能发现的超微结构．同时也运用扫描近场光学显微镜(Scanning Near field Optical Microscopy，SNOM)对淋巴细胞进行成像，观察了其对光的透射、吸收等光学性质，并对两种成像方法进行了比较．研究发现：淋巴细胞膜表面凹凸不平，分布着大量直径几十到几百纳米不等的小颗粒；淋巴细胞中央部位有自发荧光现象．结果表明，AFM和SNOM可作为进一步探讨淋巴细胞的结构与功能关系的有力工具．     

不同刺激条件下人外周血淋巴细胞超微结构与粘滞力的AFM分析

应用原子力显微镜（atomic force microscopy,AFM）探测了静息、脂多糖（LPS）或伴刀豆蛋白（ConA）活化的人外周血淋巴细胞的形态结构、超微结构及粘滞力特性。从AFM图像可知,经LPS或ConA刺激活化后的淋巴细胞比静息状态的淋巴细胞有所增大,并且表面分布着大小不一的颗粒状聚合物。利用AFM高空间分辨的力位移曲线测量系统,发现经LPS或ConA刺激活化后淋巴细胞的粘滞力是静息状态淋巴细胞的2~3倍。通过AFM探测淋巴细胞活化状态的可视化数据,可以更好地理解淋巴细胞的行为。     

Multiplex chemiluminescence microscope imaging of P16INK4A and HPV DNA as biomarker of cervical neoplasia

Classification of cervical intraepithelial neoplasia (CIN) lesions in low-grade (CIN1) or high-grade (CIN2-3) ones is crucial for optimal patient management, but current histological diagnosis on bioptic samples is often hampered by inter-observer variability. To allow objective classification, we have exploited the peculiar characteristics of chemiluminescence detection, such as high sensitivity and easy quantification of the luminescence signal, to perform sequentially in the same tissue section both an immunohistochemical quantitative detection of p16^INK4A (a protein marker of high-grade CIN lesions) and an in situ hybridization for human papillomavirus (generally accepted as a necessary but insufficient cause of cervical carcinoma). Different label enzymes (alkaline phosphatase and horseradish peroxidase) were employed in order to avoid any interference between the two assays, and quantitative chemiluminescence image analysis was used to obtain objective evaluation of sample positivity. The multiplexed method allowed detection of two complementary biomarkers and provided discrimination between different lesions (non-neoplastic, low-grade and high-grade CIN). This assay might thus represent an accurate and objective diagnostic test providing important information for counseling, selection of therapy and follow up after surgical treatment.     

Potential of Ciplukan (Physalis Angulata L.) as Source of Functional Ingredient

The aims of the study was to obtain a comprehensive data of the ciplukan as functional ingredients, whose efficacy acts as immunomodulator and antioxidant. The extraction of ciplukan was performed using ethanol and water as solvent. The total phenol content and antioxidant activity of ethanol ciplukan extract was higher than that of using water. The ciplukan leave extracts at all concentrations observed induced an increased lymphocyte cell proliferation, however, all concentrations tested showed lower stimulation index compared to the mitogen.     

基于微流控芯片的CD4+T淋巴细胞计数检测

CD4 +T淋巴细胞是人体免疫缺陷病毒(HIV)的主要感染细胞,慢性HIV感染者逐渐耗尽CD4 +T淋巴细胞,使免疫系统变弱,导致获得性免疫缺陷综合症(AIDS),因此,CD4 +T淋巴细胞的数量对HIV/AIDS的诊断和治疗至关重要。全球范围内HIV/AIDS正处于快速增长期,现有的CD4 +T淋巴细胞计数检测方法由于仪器昂贵、操作复杂、检测成本高,不利于疾病诊疗的普及与推广。为实现低成本、方便、快捷的临床检测,基于微流控芯片的CD4 +T淋巴细胞计数检测方法与技术的研究正日益受到人们的重视。本文在回顾传统CD4 +T淋巴细胞检测方法的基础上,综述、归纳了基于微流控芯片的CD4 +T淋巴细胞计数方法,在全面分析其技术特点的基础上,进一步评述了其综合性能、适用范围、及典型优缺点。最后,本文针对基于微流控芯片的CD4 +T淋巴细胞计数检测技术的发展趋势及商业化应用前景进行了讨论和展望。     

人体血液白细胞的显微光谱

采用显微光谱技术，得到了正常人及白血病患者的白细胞在可见光范围的光谱。结果显示，淋巴细胞和粒细胞的显微光谱各具特征，不同类型的白血病患者的淋巴细胞和粒细胞的显微光谱与正常人的相比也不同。结果不仅实现了白细胞的显微光谱学分类，而且可能为临床医学提供参考。     

Determination of Amino Acids in Single Human Lymphocytes after On-capillary Derivatization by Capillary Zone Electrophoresis with Electrochemical Detection

Sensitive and selective methods for the detection of amino acids (AAs) in single cells are of increasing importance. Capillary zone electrophoresis (CZE) with electrochemical detection (ED) is suitable for analysis of electroactive contents in single cells. However, most native AAs have no electroactivity. Derivatization with an electroactive tag is an attractive method. An on-capillary derivatization scheme was reported1. Naphtha- lene-2, 3-dicarboxaldehyde (NDA) can react with pri…     

放疗前后食管癌患者红细胞免疫功能和T淋巴细胞亚群的变化

目的 研究放疗前后食管癌患者红细胞免疫功能及T 淋巴细胞亚群的变化。方法 以68 例食管癌患者为研究对象（病例组）,另外选取健康献血者40 例作为对照组。以流式细胞仪检测病例组患者放疗前、接受6 周放疗后及对照组血清中T细胞亚群水平,使用受体黏附法检测红细胞免疫功能。结果 病例组患者治疗前红细胞免疫功能、血清T 细胞亚群水平均显著低于对照组（均P＜0.05）,红细胞免疫复合物花环（RBC-ICR）高于对照组（P＜0.01）;放疗后1 周,病例组的CD4＋T 细胞比例、肿瘤红细胞花环（DTER）、红细胞C3b 受体花环（RBC-C3bRR）进一步下降,而CD8＋T 细胞比例明显升高,CD4＋/CD8＋比值降低;同放疗前比较均有统计学差异（均P＜0.05）;放疗后3 个月病例组CD4＋T 细胞比例、DTER、RBC-C3bRR 逐步升高,而CD8＋T 细胞比例及RBC-ICR逐渐下降,CD4＋/CD8＋比值升高,同放疗前及放疗后1 周比较均有统计学差异（均P＜0.05）。结论 食管癌患者红细胞免疫功能及T 淋巴细胞亚群水平低下,放疗刚结束时,细胞免疫抑制作用为主导,放疗结束3 个月后,红细胞免疫功能逐渐改善、T淋巴细胞亚群逐渐恢复。放疗可有效减少肿瘤负荷,改善患者红细胞免疫及T 淋巴细胞功能。     

ICAM-1/LFA-1 interaction contributes to the induction of endothelial cell-cell separation: implication for enhanced leukocyte diapedesis

The basic route and mechanism for diapedesis has not yet to be fully defined. Here we present evidence that "cell-cell separation" between endothelial cells (ECs) may provide a route for leukocyte diapedesis. We unexpectedly found that extensive interaction between peripheral blood leukocytes and ECs that were activated by TNF-α induced the opening of EC contacts and, surprisingly, resulted in cell-cell separation. This event was specific to the intercellular adhesion molecules-1 (ICAM-1)/leukocyte function-associated antigen-1 interaction, as demonstrated by the following: (1) ICAM-1 expression correlated with increased EC contraction; and (2) the blocking of ICAM-1 selectively inhibited EC separation. Thus, we suggest that "cell-cell separation" could be a mechanism for diapedesis in situations that may require massive leukocyte infiltration.