三磷酸腺苷生物荧光液闪测定法检测活细胞计数

为建立一种检测活细胞数的方法,作者采用三磷酸腺苷(ATP)生物荧光液闪测定法检测小鼠纤维瘤细胞株L929.ATP标准曲线工作范围为10 -9～10-5mol/ml,相关系数r=0.9963(P＜0.001);当活细胞数在3×1 02～106个/ml时与发光计数值有良好的线性关系,r=0.9922(P＜0.001),变异系数 CV = 1%～3%.用ATP生物荧光液闪测定法检测活细胞数灵敏、简便、准确、重复性好,用液闪计数仪即可完成测定,有较大的实用价值.     

黄参方剂诱导胃癌癌前病变细胞凋亡和影响BCL—2蛋白表达的研究

目的：探讨黄参方剂对胃癌癌前病变细胞凋亡及相关基因表达的影响。方法：将７２例胃癌癌前变患者随机分为２组,治疗组３７例,口服黄参方剂６０ｍｌ／ｄ;对照组３５例,口服维酶素片１２片／天,疗程２月。治疗后胃镜同一部位取材,检测治疗前后细胞凋亡指数和ＢＣＬ－２蛋白的表达。采用原位末端标记法（ＴＵＮＥＬ）检测细胞凋亡,采用免疫组化检测ＢＣＬ－２蛋白的表达。结果：黄参方剂组服药前后细胞凋亡指数分别为（１９．８     

P^27的研究概况

目的：综述Ｐ＾２７的研究进展。方法：光盘检索有关献,归纳综合整理。结果：Ｐ＾２７是细胞周期的相关蛋白,Ｐ＾２７白过度表达可抑制细胞周期的进程,阻断细胞生长于Ｇ１期。结论：Ｐ＾２７是细胞周期中重要的负性调控控因子,是一种新的侯选的肿瘤抑制基因。     

运用原位DNA末端标记法研究细胞凋亡对乳腺癌预后的影响

研究临床乳腺癌标本中细胞凋亡的发生情况,评价其在预测后在的意义。方法搜集９１例浸润性乳腺癌的石蜡组织切片,运用原位ＤＮＡ断裂位点的末端标记法,检测细胞凋亡,计算凋亡细胞占肿瘤细胞的百分比,求得凋亡指数。结果本组病例细胞凋亡发生率为９１．２％,凋亡指数分为两组：０～０．２１和０．２８～０．６２,凋亡指数高低与腋淋巴结转称相关（Ｐ＜０．０１）。生存率单因素分析中,凋亡指数高的病例,无病生存率（Ｐ＝０．００９５）及总生存率（Ｐ＝０．０３４８）均优于凋亡指数低的病例。但Ｃｏｘ模型多因素分析末能指示凋亡指数是一个独立的预后指标。结论在乳腺癌组织中,细胞凋亡是一种自发现象,其发生情况各有不同,研究初步提示了细胞凋亡在乳腺癌预后中的作用,了解了凋亡和腋淋巴结转移的相关性。     

Regulatory volume response of erythrocytes exposed to a gradual and slow decrease in medium osmolality

 A sudden decrease in external medium osmolality (90 mosmol/kg) causes an immediate swelling of trout erythrocytes, followed by a regulatory volume decrease (RVD) due to activation of both a KCl cotransporter and a taurine transport pathway. Here, we determined how trout red cells respond when they are exposed to a gradual and slow decrease in medium osmolality (80 mosmol/kg at a rate of 0.7 mosmol/kg per min). Erythrocytes were unable to regulate their volume efficiently when swollen gradually and it increased continuously throughout the experimental period (120 min). As long as volume was increased slowly by 15–25%, regulatory pathways remained essentially inactivated, erythrocytes losing no significant amount of intracellular osmotically active solutes. Above this swelling threshold, a response was triggered but the quantity of solutes lost via the regulatory pathways was still not sufficient to counterbalance the continuous entry of water due to the slow and gradual decrease in medium tonicity. Received: 18 January 1999 / Received after revision: 10 February 1999 / Accepted: 11 February 1999     

Flow-mediated cell stress induction in adherent leukocytes is accompanied by modulation of morphology and phagocytic function

Leukocytes adherent to the surfaces of both vascular biomaterials and normal blood vessels experience blood flow induced shear stress. The goal of the reported studies was to investigate the effect of fluid flow on the morphology, phagocytic function and stress response induction in adherent immune cells. Shear approximating arterial, venous and intermediate levels were applied onto glass-adherent IC21 macrophages in a temperature-controlled parallel plate flow system. The results indicate that fluid flow induces a shear-dependent physiological stress response in adherent macrophages and that significant morphological changes accompany macrophage responses to shear stress. In addition, arterial flow conditions induce not only significant cell polarisation, but also enhanced phagocytic ingestion in glass-adherent IC21 macrophages. These findings suggest that blood flow induced shear stress may not only be consequent to adherent leukocyte activation, but may also be integral to the regulation of adherent leukocyte behaviour in vivo.     

雷公藤多甙对肾性蛋白尿患者外周血单个核细胞作用的研究

通过琼脂糖凝胶电泳, Ｈ Ｅ 染色和流式细胞仪检测用雷公藤多甙治疗肾性蛋白尿患者 ２ 周后外周血单个核细胞的凋亡情况。经治疗后患者外周血单个核细胞在形态上出现典型的细胞核固缩、碎裂,琼脂糖凝胶电泳显现特征性的“梯状”带,流式细胞仪上出现亚二倍体峰。提示雷公藤多甙能诱导肾性蛋白尿患者外周血单个核细胞的凋亡。     

Apoptotic cascade of neurons in the subcortical sensory relay nuclei following the neonatal infraorbital nerve transection

A terminal transferase-mediated dUTP nick end labeling (TUNEL) method was utilized for detection of neuronal death in the subcortical relay nuclei of the trigeminosensory system following the infraorbital nerve transection in newborn rats. At 18-24 h after injury, numerous TUNEL-positive profiles were found within the ventroposteromedial thalamic nucleus (VPM) contralateral to the injury, whereas the VPM on the ipsilateral side and of the age-matched normal control contained only a few profiles per section. Electron microscopy revealed that the TUNEL-positive profiles were apoptotic neurons. The ventral part of the ipsilateral brainstem sensory trigeminal nuclear complex (the nucleus principalis, and the subnuclei oralis and interpolaris) exhibited statistically significant 65-70% increase in number of apoptotic neurons compared to the contralateral side. Taken together with our previous study [T. Sugimoto, C. Xiao, H. Ichikawa, Neonatal primary neuronal death induced by capsaicin and axotomy involves an apoptotic mechanism, Brain Res. 807 (1998) 147-154], the present results demonstrated a cascade of apoptosis in the primary, secondary and tertiary order sensory neurons along the neuroaxis.     

Cellular dynamics of corneal wound re-epithelialization in the rat. I. Fate of ocular surface epithelial cells synthesizing DNA prior to wounding

The fate of ocular surface epithelial cells in response to injury of the cornea was examined. Corneal epithelial cells were labeled during DNA synthesis with [3H]thymidine 1 h prior to wounding. A 3-mm diameter epithelial defect was made in the center of the rat cornea, with the basement membrane remaining intact. Within 12 h of abrasion, labeled cells were detected in the regenerating surface. At 18 h, there was a 2.7- and 17-fold increase of labeled basal and suprabasal cells, respectively, in the epithelium adjacent to the wound, and at 24 and 30 h there was an excessive number of cell layers (up to 7) at the margin of the abrasion. Re-epithelialization progressed as a gradient of cell layers that became diminished towards the center of the wound. Completion of layers 1, 2, 3, and 4 were recorded at 24, 30, 36, and 72 h, respectively. No changes in the labeling index of the limbus or conjunctiva were noted. These results suggest that processes of centripetal and vertical migration, as well as events related to cell division, in the uninjured corneal surface are not impeded by wounding of the corneal epithelium. However, wound healing appears to require cells with a basal phenotype, presumably because of this cell type's migratory capability.