阿司匹林、肝素对血小板活化及表达CD40L的影响

目的探讨血小板活化与表达CD40L的关系以及阿司匹林、肝素对此过程的影响.方法体外分离健康人血小板,经不同浓度二磷酸腺苷(ADP)、凝血酶诱导后,应用流式细胞术测定血小板活化指标P选择素和炎性标志CD40L表达水平,观察二者随诱导时间延长的变化过程,并分析阿司匹林、普通肝素、低分子肝素对血小板活化及表达CD40L的影响.结果 ADP、凝血酶均呈浓度依赖方式增加血小板P选择素、CD40L表达,二者表达水平随诱导时间延长而同步增减,呈显著正相关(P<0.05).阿司匹林(2.5 μg/ml)对ADP(4 μmol/L)和凝血酶(1U/ml)诱导的血小板活化及CD40L表达无任何影响(P>0.05);普通肝素(2.5U/ml)和低分子肝素(2.5U/ml)单独或与阿司匹林合用,均能极显著地抑制凝血酶诱导的血小板活化及CD40L表达(P＜0.001),但对ADP的诱导过程无影响(P>0.05).结论炎性介质CD40L可表达在活化血小板表面,在多种诱导剂存在的情况下,阿司匹林及肝素能部分抑制血小板的活化及CD40L表达.     

不稳定性心绞痛患者活化血小板表达细胞分化抗原40配体及其与可溶性黏附分子水平的关系

目的 :探讨不稳定性心绞痛 (UA)患者体内血小板活化及表达细胞分化抗原 40配体 (CD40L)与其血清可溶性黏附分子水平的关系。　　方法 :流式细胞术测定 3 8例UA患者 (UA组 ) ,2 2例稳定性心绞痛 (SA)患者 (SA组 )及 2 1例对照者 (对照组 )血小板P 选择素 (CD62P)及CD40L表达 ,并与体外诱导的健康人血小板活化程度及CD40L表达水平进行比较。同时用ELISA法测定 3组研究对象的血清可溶性细胞间黏附分子 1(sICAM 1)和血管细胞黏附分子 1(sVCAM -1)浓度 ,分析UA患者血小板表达CD40L与其可溶性黏附分子水平的关系。　　结果 :UA组血小板CD62P、CD40L表达显著高于SA组及对照组 (P <0 0 1) ,达到了健康人血小板在体外经中等浓度诱导剂刺激时CD62P、CD40L的表达水平。同时 ,UA患者血清sICAM 1、sVCAM 1浓度也显著高于SA组及对照组 ,并分别与其血小板表达CD40L呈显著正相关 (γ =0 483 ,γ =0 5 16,P <0 0 5 )。　　结论 :UA患者体内血小板过度激活的同时可表达炎性介质CD40L ,并与其血清可溶性黏附分子水平升高有关。     

细胞黏附分子-1在急性髓性白血病细胞与内皮细胞黏附中的作用

目的　检测急性髓性白血病 (AML)细胞与内皮细胞的黏附及细胞黏附分子 1(ICAM 1)及其配体淋巴细胞功能相关抗原 1(LFA 1)在黏附中的作用。方法　观察AML细胞与静止内皮细胞和肿瘤坏死因子α(TNFα)激活的内皮细胞的黏附 ;AML细胞与内皮细胞混合培养 2 4h的黏附 ;正常中性粒细胞与AML细胞培养上清作用 2 4h后的内皮细胞的黏附 ;流式和ELISA方法检测AML细胞培养上清作用后内皮细胞ICAM 1及可溶性ICAM 1的表达 ;并用ICAM 1和LFA 1的抗体进行阻滞黏附试验。结果　AML细胞与静止的内皮细胞黏附较少 (2 4 33± 2 87) % ,AML细胞与TNFα激活的内皮细胞的黏附 ,与内皮细胞混合培养 2 4h后的黏附以及正常中性粒细胞与AML细胞培养上清作用后内皮细胞的黏附明显增加 ,分别为 (81 87± 4 0 8) % ,(82 0 6± 7 0 5 ) % ,(83 99± 3 86 ) % (n =2 1,P <0 0 0 1) ;静止的内皮细胞ICAM 1及可溶性ICAM 1的表达分别为 (5 5 81± 4 11) %和 (0 839± 0 2 36 )μg/L ;AML细胞培养上清作用后内皮细胞ICAM 1及可溶性ICAM 1的表达明显增加 ,分别为 (6 5 36±5 97) %和 (1 4 2 4± 0 4 6 9) μg/L(n =2 1,P <0 0 5 ) ;用ICAM 1及LFA 1的抗体进行黏附阻滞后 ,AML细胞与TNFα激活的内皮细胞的黏附下降为 (2 0 12±     

血小板在动脉粥样硬化形成和发展中的作用

近年来的一些研究显示血小板可作为“炎症细胞” ,活化或释放的炎症介质直接参与动脉粥样硬化的形成和发展。急性冠状动脉综合征被称为动脉硬化血栓性疾病 ,因此抗血小板治疗在ACS发病中起十分重要的作用。1　血小板参与动脉粥样硬化形成和发展的机制血小板活化后释放多种炎症介质 ,它们可以 :(1)促进血小板黏附和聚集 ;(2 )促进血小板与白细胞黏附并激活后者促使白细胞与内皮细胞黏附和白细胞向血管内膜迁移 ;(3)诱导血管平滑肌细胞的迁移和增生 ;(4)活化的血小板能够表达CD15 4 ,CD15 4激活后可显著上调血管细胞黏附分子 1和细胞间黏附…     

Pin1对oxLDL诱导内皮细胞炎症反应的作用及机制研究

目的探讨肽基脯氨酰顺反异构酶1(Pin1)对氧化低密度脂蛋白(oxLDL)诱导内皮细胞炎症反应的影响。方法人脐静脉内皮细胞系予以不同浓度oxLDL干预12、24、48h;并预先转染Pin1 siRNA或给予STAT3抑制剂后,采用50μg/ml oxLDL干预内皮细胞15min或24h,检测内皮细胞中血管细胞黏附分子1(VCAM-1)、Pin1蛋白表达水平,STAT3活化水平和单核-内皮细胞黏附情况。结果与对照组相比,oxLDL 50μg/ml组和100μg/ml组细胞内VCAM-1、Pin1蛋白表达明显增加(P0.05); oxLDL50μg/ml干预12、24、48h后,细胞内VCAM-1、Pin1蛋白表达亦明显增加(P0.05)。与对照组相比,oxLDL50μg/ml干预15min后,胞内p-STAT3明显增加(P0.05);oxLDL50μg/ml干预24h后,内皮细胞上黏附的单核细胞数量明显增多(P0.05)。与oxLDL组相比,Pin1 siRNA干扰后显著降低了oxLDL诱导的内皮细胞p-STAT3水平、VCAM-1蛋白表达水平(P0.05),也明显减少了oxLDL诱导的单核细胞黏附水平(P0.05)。此外,STAT3抑制剂显著降低了oxLDL诱导的内皮细胞VCAM-1蛋白表达与单核细胞黏附水平(P0.05)。结论 Pin1通过上调STAT3信号通路活化水平促进oxLDL诱导的内皮细胞炎症反应。     

他汀类药对人脐静脐内皮细胞细胞间黏附分子—1表达的影响

目的　观察氟伐他汀 (Flu)和辛伐他汀 (Sim)对肿瘤坏死因子 (TNF)α诱导的人脐静脉内皮细胞 (HUVEC)细胞间黏附分子 (ICAM) 1表达的影响 ,探讨他汀类药物可能的非调脂抗动脉粥样硬化作用。方法　体外培养HUVEC ,加TNFα (10 0U/ml)及 1× 10 -8～ 1× 10 -5mol/L浓度Flu或Sim ,共孵育 6、12和 2 4h ,采用细胞ELISA、流式细胞技术和逆转录 聚合酶链反应测定ICAM 1蛋白及mRNA水平表达。结果　Flu和Sim皆对TNFα诱导的ICAM 1蛋白表达及ICAM 1mRNA表达有抑制作用 ,呈浓度依赖性和时间依赖性特征。结论　Flu和Sim抑制TNFα诱导的HUVECICAM 1表达 ,可能对阻止血单核细胞向血管内皮细胞募集和黏附、延缓动脉粥样硬化的发生和发展有一定作用。     

HCV刺激人脐静脉内皮细胞发生动脉粥样硬化的机制

目的以HCV体外刺激人脐静脉内皮细胞(HUVECs)为模型,探讨HCV感染致动脉粥样硬化发生的机制。方法采用1. 0 MOI HCV病毒颗粒刺激HUVECs,CCK8检测细胞增殖;流式细胞仪检测细胞凋亡及周期;划痕实验及单核内皮黏附实验评估HCV对HUVECs迁移及黏附能力的影响;荧光定量PCR及Western blot检测HCV刺激HUVECs炎症因子及内皮损伤因子的表达。2组间比较采用两独立样本t检验,多组间比较采用方差分析,进一步两两比较采用LSD-t检验。结果与对照组比较,HCV对HUVECs的生长增殖、细胞凋亡及周期无明显影响(P值均 0. 05)。HCV刺激抑制了HUVECs的迁移能力,而增强其黏附能力。与对照组比较,HCV刺激促进内皮细胞炎症因子IL-6、IL-1β以及趋化因子CXCL10、单核细胞趋化蛋白-1 mRNA水平升高(t值分别为-10. 155、-12. 048、-5. 025、-20. 116,P值均0. 05)及蛋白表达增加(F值分别为2541. 739、4806. 490、477. 608、501. 380,P值均0. 001)。HCV刺激导致HUVECs内皮损伤因子内皮素-1、血管内皮生长因子以及黏附分子—细胞间黏附分子-1、血管细胞黏附因子-1表达上调(t值分别为-4. 530、-4. 497、-7. 692、-7. 449,P值均0. 05)。结论 HCV可以引起内皮细胞炎症改变和功能障碍,影响动脉粥样硬化的发生。     

男性吸烟者细胞黏附分子和相关细胞因子水平的观察

目的 :观察男性吸烟者血清可溶性黏附分子浓度和血细胞黏附分子表达的变化。　　方法 :选血压、血脂、体重指数正常的健康青年男性吸烟者 2 8名为吸烟组 ,5分内吸烟 1支 ,观察吸烟前及吸烟后 5分时血清可溶性E 选择素 (sE Selectin)、可溶性细胞间黏附分子 (sICAM ) 1、可溶性血管细胞黏附分子 (sVCAM )浓度的变化 ,血小板P 选择素、白细胞L 选择素、细胞间黏附分子 (ICAM ) 1、ICAM 3的表达 ,同时测定血清白细胞介素 1β、白细胞介素 8、肿瘤坏死因子的含量 ;吸烟指数为每日吸烟支数与吸烟年限的乘积。选择无吸烟史的健康男性 2 2名作为对照组。　　结果 :吸烟组吸烟前血清sE Selectin、sICAM 1高于对照组 ,有显著性差异 (P <0 0 1及P <0 0 2 ) ,吸烟前后无明显变化 ;P 选择素阳性血小板百分率吸烟组吸烟前高于对照组 ,有极显著性差异 (P <0 0 0 1) ,吸烟前后无明显变化 ;吸烟指数与P 选择素阳性血小板百分率及sE Selectin、sICAM 1相关 (γ值分别为 0 5 10、0 43 3及 0 42 9,P <0 0 1、P <0 0 5及P <0 0 5 )。白细胞L 选择素、ICAM 1及ICAM 3阳性细胞百分率及平均荧光强度吸烟组吸烟前与对照组无显著差异 ,吸烟刺激后仍无明显改变。血清细胞因子的含量两组差异不明显 ,吸烟后亦无明显变     

活化血小板诱导内皮细胞表达黏附分子

血管内皮细胞释放黏附分子 ,促使循环单核细胞、淋巴细胞向局部黏附聚集 ,由此引发的炎症反应可造成冠状动脉内不稳定斑块发生破裂 ,继发血小板活化与血栓形成、并导致急性心肌缺血事件的发生[1 ] 。新近研究发现 ,血小板在其活化的同时尚可表达炎性介质ＣＤ1 54分子[2 ] 。提示除了参与血栓形成外 ,活化血小板也有可能通过ＣＤ1 54 ＣＤ4 0 信号途径主动介导炎症反应 ,但这一假设尚未被证实。为此 ,我们对血小板活化与其表达ＣＤ1 54间的确切关系进行了探讨。一、材料与方法1 .血小板的制备及诱导活化 :静脉血取自健康献血者及志愿者 (1 6…     

CD40配体高表达与不稳定型心绞痛之间的关系

为了探讨不稳定型心绞痛患者外周血单核细胞表达CD40配体及血清可溶性CD40配体变化的临床意义。应用间接免疫荧光流式细胞术和双抗夹心酶联免疫测定法分别对正常对照组 16例、稳定型心绞痛 2 0例、不稳定型心绞痛 2 0例患者血单核细胞表达CD40配体及血清可溶性CD40配体水平进行检测。并观察血清可溶性CD40配体与冠状动脉病变程度的关系。结果发现 ,(1)不稳定型心绞痛组血单核细胞表达CD40配体明显较稳定型心绞痛组和对照组高 (P <0 .0 1) ,稳定型心绞痛组与对照组相比无差异 (P >0 .0 5 )。 (2 )不稳定型心绞痛组血清可溶性CD40配体水平明显较稳定型心绞痛组和对照组高 (P <0 .0 1) ,而稳定型心绞痛组与对照组相比亦有明显差异 (P <0 .0 5 )。 (3)经皮冠状动脉腔内成形术后血清可溶性CD40配体明显高于术前 (P <0 .0 1) ,但血单核细胞表达CD40配体无差异 (P >0 .0 5 )。 (4 )血清可溶性CD40配体水平与冠状动脉病变的复杂狭窄数相关 (r=0 .5 4,P <0 .0 1) ,而与狭窄的程度和范围无关。此结果提示 ,血清可溶性CD40配体升高对冠状动脉斑块的不稳定或破裂起重要作用 ,且可能是冠状动脉病变的活动性标志物。     

Three layer functional model and energy exchange concept of aging process

Relying on a certain degree of abstraction, we can propose that no particular distinction exists between animate or living matter and inanimate matter. While focusing attention on some specifics, the dividing line between the two can be drawn. The most apparent distinction is in the level of structural and functional organization with the dissimilar streams of ‘energy flow’ between the observed entity and the surrounding environment. In essence, living matter is created from inanimate matter which is organized to contain internal intense energy processes and maintain lower intensity energy exchange processes with the environment. Taking internal and external energy processes into account, we contend in this paper that living matter can be referred to as matter of dissipative structure, with this structure assumed to be a common quality of all living creatures and living matter in general. Interruption of internal energy conversion processes and terminating the controlled energy exchange with the environment leads to degeneration of dissipative structure and reduction of the same to inanimate matter, (gas, liquid and/or solid inanimate substances), and ultimately what can be called ‘death.’ This concept of what we call dissipative nature can be extended from living organisms to social groups of animals, to mankind. An analogy based on the organization of matter provides a basis for a functional model of living entities. The models relies on the parallels among the three central structures of any cell (nucleus, cytoplasm and outer membrane) and the human body (central organs, body fluids along with the connective tissues, and external skin integument). This three-part structural organization may be observed almost universally in nature. It can be observed from the atomic structure to the planetary and intergalactic organizations. This similarity is corroborated by the membrane theory applied to living organisms. According to the energy nature of living matter and the proposed functional model, the decreased integrity of a human body's external envelope membrane is a first cause of the structural degradation and aging of the entire organism. The aging process than progresses externally to internally, as in single cell organisms, suggesting that much of the efforts towards the restoration and maintenance of the mechanisms responsible for structural development should be focused accordingly, on the membrane, i.e., the skin. Numerous reports indicate that all parts of the human body, like: bones, blood with blood vessels, muscles, skin, and so on, have some ability for restoration. Therefore, actual revival of not only aging tissue of the human body's membrane, but the entire human body enclosed within, with all internal organs, might be expected. We assess several aging theories within the context of our model and provide suggestions on how to activate the body's own anti-aging mechanisms and increase longevity. This paper presents some analogies and some distinctions that exist between the living dissipative structure matter and inanimate matter, discusses the aging process and proposes certain aging reversal solutions.     

Unusual responses of nocturnal pineal melatonin synthesis and secretion to swimming: Attempts to define mechanisms

Abstract: The effect of swimming at night on rat pineal melatonin synthesis was compared with that of light exposure at night. Rats were forced to swim at 0030 hr (lights out at 2000 hr) and sacrificed by decapitation 15 and 30 min later, immediately after swimming. Other groups of animals were exposed to white light (650μW/cm²) for 15 and 30 min at same time. Swimming caused a rapid and highly significant drop in the melatonin content in the pineal gland; however, the activity of N-acetyltransferase (NAT), the supposed rate limiting enzyme in the melatonin production, was not changed. Despite the drop in pineal melatonin levels, serum concentrations of the indole remained elevated in the rats that swam. In contrast, melatonin levels in the pineal and serum of light exposed rats fell precipitously, accompanied by a significant suppression of NAT activity. Since we anticipated that the strenuous exercise associated with swimming may induce release of artrial natriuretic peptide (ANP) from the heart, which in turn could cause the release of pineal melatonin, in a second study we injected physiological saline intravenously to stretch the cardiac muscle and release ANP. Three milliliters of normal saline was injected during the day into the jugular vein of anesthetized rats that were pretreated with isoproterenol to stimulate pineal melatonin production. Animals were killed 15 min after the saline injection, and pineal NAT activity and pineal melatonin levels were measured. The saline injections caused no alteration in the elevated levels of either NAT or melatonin. These data suggest that the disparity in pineal NAT activity (which was high) and pineal melatonin (which was low), in animals swum at night, may not be caused by ANP which is released during strenuous exercise such as swimming.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Melatonin and photoperiodic time measurement: Seasonal breeding in the sheep

Abstract: Well-established circadian physiology supports the view that photoperiodic time measurement utilizes the coincidence between the presence of light and a photosensitive phase of a 'biological clock' to alter reproductive status—the so-called external coincidence model of seasonal breeding. In this review, we examine the mechanism whereby photoperiod interacts with presumed suprachiasmatic nuclei activity to allow endogenous melatonin to normally synchronize reproductive activity to the optimal time of year. The Romney Marsh sheep is particularly explored as an experimental model. It is suggested that the on/off activity of seasonal reproduction may be a robust mechanism able to be predictably manipulated by the judicious use of the light/dark cycle and exogenous melatonin, but firmly based on circadian principles.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.