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1.
在相当长的一段时间里,各种自动白细胞识别方法已被广泛的应用。其中有一种方法应用了VCS技术。这种方法即是本文将讨论的内容。一种专用试剂能够防止细胞受染色溶剂或细胞化学试剂的影响而改变。细胞的一些原始特性,诸如白细胞的大小,内部的物理、化学特性,以及各个细胞特殊的表面结构特性可基本保持不变。VCS方法是同时运用体积分析(AnalysisofVolume),传导系数(Conductivity)和激光光散射(Laser-light-Scattering)三种技术,得到每个白细胞几乎处于原始状态的多维记录  相似文献   

2.
重组人粒细胞集落刺激因子 (rhG CSF)是应用基因重组技术生产的糖蛋白 ,能有效调解造血祖细胞的粒系增殖分化 ,可用于治疗恶性肿瘤放疗和化疗引起的白细胞减少症[1] 。我们自 1998年 9月~ 1999年 11月使用国产rhG CSF治疗肿瘤化疗所致白细胞减少症取得了满意效果。1 材料及方法14例患者中 ,男性 9例 ,女性 5例 ,平均年龄 5 3岁 (32~71岁 )。病理均证实为恶性肿瘤 ,其中小细胞肺癌 4例 ,化疗方案CE(CBP ,VP16 ) ;非小细胞肺癌 5例 ,化疗方案NP(NVB ,DDP) ;胃腺癌 3例 ,化疗方案EFP(VP16 ,5FU ,DDP) …  相似文献   

3.
本文对正常孕妇、妊娠高血压综合征(PIH)患者和经青心酮(DHAP)治疗的PIH患者等共24例,应用组织化学分析方法观察胎盘血管内皮细胞(VEC)和平滑肌细胞(VSMC)内一氧化氮合酶(NOS)活性的变化。结果表明:正常孕妇胎盘VEC和VSMC内NOS活性较高;PIH胎盘VEC和VSMC内NOS活性明显减弱,并伴有组织和细胞的形态学损伤;经DHAP治疗后的PIH胎盘VEC和VSMC细胞NOS活性较未经DHAP治疗者明显增加,其组织和细胞损伤也减轻。本研究结果提示胎盘内VEC和VSMC细胞的NOS减少可能与PIH的发生和/或发展有关,青心酮治疗PIH的作用可能与DHAP促进胎盘VEC和VSMC内一氧化氮(NO)合成有关。  相似文献   

4.
利用HSV—TK基因治疗肝癌的离体研究   总被引:2,自引:1,他引:1  
利用DNA重组技术,将HSV-TK基因克隆至逆转录病毒载体(XM-6/TK)。经PA317细胞包装后,转染人肝癌细胞HepG2。结果显示,XM-6/TK转染HepG2细胞与野生型相比,其生长特点和细胞形态未有改变。给予抗病毒药物-环氧鸟苷(Ganciclovir,GCV)后,转染细胞生长受到严重抑制,细胞数量明显降低。细胞学检查证实,GCV处理后,转染细胞的死亡率显著增加。本结果提示,应用HSV-TK基因治疗肝癌可能为一种治疗肿瘤新的方法  相似文献   

5.
血管平滑肌细胞增殖与Cdk抑制蛋白p27的表达   总被引:4,自引:1,他引:4  
Yuan Y  Xu DL  Liu YL  Jia MY 《生理学报》1999,51(3):285-290
p27蛋白是细胞周期素依赖性激酶(Cdk)抑制蛋白家族中的一种,主要对外部促进或抑制细胞增殖的信号起反应。本研究应用流式细胞仪(FCM)双标记的方法观察血管紧张素Ⅱ(AngⅡ)、血管加压素(AVP)和血小板源生长因子(PDGF)对血管平滑肌细胞(VSMCs)细胞周期百分比和p27蛋白表达量的影响。静止状态培养的VSMCs加入AngⅡ,AVP,PDGFBB后,在不同时间收集细胞,用碘化丙啶(PI)标记细胞DNA,以确定细胞所处的周期。用p27蛋白的单抗和标记了FITC的二抗标记细胞,通过流式细胞仪测定被激发出的荧光量来确定细胞p27蛋白表达的相对量。结果显示,AngⅡ刺激VSMCs增生,其蛋白含量增加了436%(P<001),但不抑制p27蛋白的表达;AVP可轻度抑制p27的表达,有轻度促进VSMCs增殖和增生的作用(P<005);PDGF明显抑制p27的表达,引起细胞增殖。本研究结果提示,p27蛋白抑制VSMCs通过G1期进入S期,是抑制VSMCs增殖的重要调节因子。  相似文献   

6.
应用原位分子杂交及免疫组分方法,使用地高辛标记HCV5’非编码区探针及抗HCV NS3区C33c单克隆抗体,对35例人原发性肝内胆管细胞癌(PIC)和癌旁肝组织的HCV RNA及其NS3抗原进行检测结果发现HCV RNA在PIC中的阳性率为83%,HCV RNA定位于癌细胞浆中,个别病例并在淋巴细胞、肝窦内皮细胞和枯否氏细胞中发现阳性信号。HCV NS3区C33c抗原在PIC的阳性率为89%,阳性  相似文献   

7.
本研究用培养大鼠主动脉血管平滑肌细胞(VSMCs),结果如下:(1)用生物活性检测法发现VSMCs无血清条件培养液可刺激巨噬细胞集落形成,其作用能被抗巨噬细胞集落刺激因子(MCSF)抗体抑制;(2)用免疫细胞化学技术证明VSMCs存在MCSF受体;(3)用Northern blot技术证明VSMCs有MCSF及其受本的mRNA表达,血清刺激使两者表达明显增强。本研究首次报道了培养大鼠主动脉VSMC  相似文献   

8.
猪瘟病毒在PK细胞和MPK细胞中繁殖过程的研究   总被引:13,自引:2,他引:13  
以猪瘟病毒疫苗Thiverval株(T株)为实验材料,研究该病毒株在PK15细胞中增殖的基本特性与规律。在PK15细胞中,猪瘟病毒T株在感染后12h即可检测到子代病毒粒子。接毒后48h,几乎所有的细胞都被病毒感染;到60h,释放到培养液中有活性的病毒粒子达到最高峰,为107TCID50/mL。培养液中的病毒粒子在37℃半寿期只有3个小时。同时,建立了MPK细胞CSFVT株的感染模式,其CSFV的滴度可达108TCID50/mL。在此基础上,用抗CSFV包膜蛋白E2和非结构蛋白p120的单克隆抗体显示了病毒在细胞中增殖的部位,进而应用电镜技术观察到成熟的病毒粒子及可能处在不同发育阶段的子代病毒粒子  相似文献   

9.
王镇  陆宇  丁明孝   《微生物学通报》2000,27(2):79-84
猪瘟病毒(CSFV)能在多种体外培养细胞中增殖,却不使细胞产生病变(CPE)。使用原代细胞增殖CSFV弱毒疫苗。结果显示:病毒的增殖能够增加原代牛睾丸细胞的传代次数和维持时间。因此,CSFV疫苗一产上能够在接毒后多次收获病毒。此外,CSFV弱毒株还能够刺激体外培养的兔巨噬细胞增殖,使形成致密的单细胞层。使用传代细胞PK15样殖病毒,经流式细胞术检测发现CSFV弱毒的增殖不仅不改变PK15细胞的增殖  相似文献   

10.
应用RNA印迹分析和亚硝酸盐含量测定检查脂多糖(LPS)对大鼠血管平滑肌细胞(VSMC)一氧化氮合酶(NOS)基因表达及NO合成的影响,用3H-TdR参入实验观察LPS对细胞DNA合成的影响.结果表明,LPS在诱导VSMCiNOSmRNA表达和促进NO合成的同时,抑制VSMCDNA合成.证明LPS的作用与其浓度和作用时间有关  相似文献   

11.
In contrast to acute preparations such as the exteriorized mesentery or the cremaster muscle, chronically instrumented chamber models allow one to study the microcirculation under "physiological" conditions, i.e., in the absence of trauma-induced leukocyte rolling along the venular endothelium. To underscore the importance of studying the naive microcirculation, we implanted titanium dorsal skinfold chambers in hamsters and used intravital fluorescence microscopy to study venular leukocyte rolling in response to ischemia-reperfusion injury or extracorporeal blood circulation. The experiments were performed in chambers that fulfilled all well-established criteria for a physiological microcirculation as well as in chambers that showed various extents of leukocyte rolling due to trauma, hemorrhage, or inflammation. In ideal chambers with a physiological microcirculation (<30 rolling leukocytes/mm vessel circumference in 30 s), ischemia-reperfusion injury and extracorporeal blood circulation significantly stimulated leukocyte rolling along the venular endothelium and, subsequently, firm leukocyte adhesion. In contrast, both stimuli failed to elicit leukocyte rolling in borderline chambers (30-100 leukocytes/mm), and in blatantly inflamed chambers with yet higher numbers of rolling leukocytes at baseline (>100 leukocytes/mm), we observed a paradoxical reduction of leukocyte rolling after ischemia-reperfusion injury or extracorporeal blood circulation. A similar effect was observed when we superfused leukotriene B4 (LTB4) onto the chamber tissue. The initial increase in leukocyte rolling in response to an LTB4 challenge was reversed by a second superfusion 90 min later. These observations underscore 1) the benefit of studying leukocyte-endothelial cell interaction in chronically instrumented chamber models and 2) the necessity to strictly adhere to well-established criteria of a physiological microcirculation.  相似文献   

12.
J E Valinsky  T G Easton  E Reich 《Cell》1978,13(3):487-499
We have reported (Easton, Valinsky and Reich, 1978) that merocyanine 540 (MC 540) specifically stains a variety of living excitable cells, but not nonexcitable cells. This paper describes the exceptional permeability to MC 540 of leukemic leukocytes and immature hemopoietic precursor cells. We have used fluorescence microscopy and uptake of radioactive dye to study MC 540 staining of peripheral blood leukocytes from 80 leukemic and 34 normal individuals; leukemic leukocytes stain, whereas normal leukcytes do not. The leukocyte staining reaction differs from that previously described for excitable cells since it is independent of the ionic composition of the staining medium, kinetically complex, enhanced by light, enhanced by oxygen and essentially irreversible. Virtually all circulating nucleated cells from leukemic individuals are stained to approximately the same extent, and there is no qualitative or quantitative distinction between the various forms of leukemia. We have also found that MC 540 interacts with granulopoietic colony-forming cells (CFU-C) and with spleen colony-forming cells derived from mouse bone marrow (CFU-S). We cannot as yet identify a specific property of leukocyte plasma membranes that determines MC 540 permeability; since changes in MC 540 uptake appear to be correlated with cellular maturation during normal hemopoiesis, the retention of staining by leukemic cells, some of which appear morphologically normal, may indicate of failure in membrane maturation during leukemic blood cell development.  相似文献   

13.
Summary A rapid, semiautomated system to quantitate and analyze leukocyte shape and locomotion was developed. Video images of moving leukocytes were obtained using a Vidicon camera mounted on a Nikon phase microscope. The video signal was either inputted directly, or indirectly via a video cassette recorder, to a Datacube video analog-digital, digital-analog converter. A Digital Equipment Corporation LSI 11/23 computer using the RT-11/TSX-Plus operating system and computer programs written in FORTRAN and MARCO assembly language permitted image segmentation, image display, and calculation of position, speed, direction of movement and orientation of each leukocyte at 10 s intervals. These data were stored on a winchester disk for subsequent evaluation of the leukocyte orientation, speed and direction of movement using statistical and graphical methods. The reproducibility of measurements made with the video system was tested by comparison with manual measurements; a correlation coefficient of 0.998 was obtained for the two methods. Rates of chemokinesis were then determined for unstimulated and chemokinetically stimulated polymorphonuclear leukocytes (PMNs) and found to average 12.8 m/min and 18.1 m/min, respectively. The high speed, ease of data analysis, and potential for multiparameter evaluation makes this system useful for directly evaluating leukocyte locomotion.In honour of Prof. P. van Duijn  相似文献   

14.
Digital pathology and microscope image analysis is widely used in comprehensive studies of cell morphology. Identification and analysis of leukocytes in blood smear images, acquired from bright field microscope, are vital for diagnosing many diseases such as hepatitis, leukaemia and acquired immune deficiency syndrome (AIDS). The major challenge for robust and accurate identification and segmentation of leukocyte in blood smear images lays in the large variations of cell appearance such as size, colour and shape of cells, the adhesion between leukocytes (white blood cells, WBCs) and erythrocytes (red blood cells, RBCs), and the emergence of substantial dyeing impurities in blood smear images. In this paper, an end‐to‐end leukocyte localization and segmentation method is proposed, named LeukocyteMask, in which pixel‐level prior information is utilized for supervisor training of a deep convolutional neural network, which is then employed to locate the region of interests (ROI) of leukocyte, and finally segmentation mask of leukocyte is obtained based on the extracted ROI by forward propagation of the network. Experimental results validate the effectiveness of the propose method and both the quantitative and qualitative comparisons with existing methods indicate that LeukocyteMask achieves a state‐of‐the‐art performance for the segmentation of leukocyte in terms of robustness and accuracy .  相似文献   

15.
An optimized procedure is described for isolation and highefficiency radiolabeling of leukocytes using 111In-oxine. The chief advantages over conventional methods include virtually no loss of leukocytes during washing and separation steps; a significant reduction in the time required to prepare leukocytes for radiolabeling compared to non-hemolytic preparations; a 28% increase in the average labeling efficiency obtained using 111In-oxine; >95% cell viability as measured by the trypan blue exclusion test; elimination of contaminating red blood cells from the leukocyte pellet prior to labeling; and 80% survivability at 15 min post injection (measured as per cent of blood activity on leukocyte fraction).  相似文献   

16.
A method has been described for removing leukocytes and thrombocytes from preserved ACD blood and from packed red cells of the same type of blood. A system has been used comprising two connected plastic bags model "Fenval". The leukocyte and thrombocyte removal is performed by two consecutive procedures, each comprising centrifugation with subsequent fast sedimentation of erythrocytes. No filters are needed. 97.3 +/- 3.04% leukocyte removal and 96.90 +/- 5.96% thrombocyte removal as regards the preserved blood and 95.65 +/- 8.70% leukocyte removal and 98.44 +/- 0.70% thrombocyte removal as regards the packed red cells have been achieved.  相似文献   

17.
Petri B  Bixel MG 《The FEBS journal》2006,273(19):4399-4407
The recruitment of leukocytes from the circulation into tissues requires leukocyte migration through the vascular endothelium. The mechanisms by which leukocytes attach and firmly adhere to the endothelial cell surface have been studied in detail. However, much less is known about the last step in this process, the diapedesis of leukocytes through the vascular endothelium. This minireview focuses on the interactions between leukocyte and endothelial cell adhesion molecules that are important during leukocyte extravasation. In the past few years a series of endothelial cell surface and adhesion molecules have been identified that are located at endothelial cell contacts and found to participate in leukocyte diapedesis. These junctional cell adhesion molecules are believed to have an active role in controlling the opening and closure of endothelial cell contacts to allow the passage of leukocytes between adjacent endothelial cells. Alternatively, leukocytes can cross the endothelium at nonjunctional locations, with leukocytes migrating through a single endothelial cell. Further work is clearly needed to understand, in greater detail, the molecular mechanisms that allow leukocytes to cross the endothelium via either the paracellular or the transcellular pathway.  相似文献   

18.
Summary The effect of purified protein derivative (PPD) on human granulopoiesis was studied in an in vitro semisolid culture system of human bone marrow in which PPD was incorporated into the leukocyte feeder layers. We observed that preincubation of the feeder layers with PPD was necessary to induce a significant rise of agar culture colony-forming units (CFU-c) with a maximum of 3 days' preincubation and a dose of 200 g for 10 6 leukocytes. A similar effect was obtained when a conditioned medium from PPD-stimulated leukocytes was used instead of feeder layers. We have found a significant correlation between the skin test response of the leukocyte donors to PPD and the colony-stimulating activity of their leukocytes exposed to PPD: these results suggest that PPD could stimulate human granulopoiesis by an indirect effect on CSF-producing mononuclear cells.  相似文献   

19.
Glycogen synthase from human and bovine polymorphonuclear leukocytes was purified to homogeneity. Rabbit antisera were raised against the two glycogen synthases and used for immunochemical analysis. Western blotting analysis showed that the subunit of glycogen synthase in crude homogenates of human and bovine leukocytes in both cases has an Mr of 85 000. The existence of a cross-reactivity between the two enzymes and the corresponding antisera demonstrates immunological similarities between bovine and human leukocyte glycogen synthase. In addition, both antisera recognize glycogen synthase in crude cellular extracts from rabbit and rat liver and from skeletal muscle. Leukocyte glycogen synthase, therefore, cannot be classified as either muscle (M-type) or liver (L-type) glycogen synthase and our results do not support the proposed immunochemical distinction between M- and L-type glycogen synthase.  相似文献   

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