Mast cells express connexins on their cytoplasmic membrane

BACKGROUND: Because of the close association between mast cells and fibroblasts in the microenvironment and the importance of connexins (Cxs) in fibroblast communication with other cells, we hypothesized that mast cells also express Cxs, allowing them to similarly communicate with other cells through gap junctions. OBJECTIVES: We sought to identify the expression of Cxs (particularly Cx43, Cx32, and Cx26) by murine mast cells. METHODS: The expression of Cxs was studied by RT-PCR, Northern blot analysis, Western blot analysis, flow cytometry, and confocal laser scanning microscopy. RESULTS: In this report we demonstrate that murine bone marrow cultured mast cells and the growth factor-independent murine mast cell line C57, express Cx43 and Cx32 as assessed by RT-PCR, Northern blot analysis, Western blot analysis, and flow cytometry, but do not express Cx26. We also show, by confocal laser scanning microscopy, that Cx43 localizes to the cytoplasmic membrane of mast cells in a pattern similar to that seen in fibroblasts. CONCLUSIONS: Mast cells express Cx43 and Cx32, and Cx43 is associated with the cytoplasmic membrane, suggesting that mast cells have the potential to communicate with other cells in their microenvironment in part through gap junctions.     

Fibrinogen polymorphisms TaqI, HaeIII and BclI are not associated with a higher risk of deep vein thrombosis

High fibrinogen is recognised as a risk factor for atherosclerosis. It seems that high fibrinogen is also a risk factor for deep vein thrombosis (DVT). It has been shown that certain polymorphisms in fibrinogen genes can influence the fibrinogen level. In this study, fibrinogen levels and the frequency of the polymorphisms TaqI, HaeIII and BclI were studied in 114 patients with DVT and 244 healthy subjects. In non-smokers, fibrinogen levels above 5 g/l were associated with an increased risk of DVT (odds ratio 3.3, 95% confidence interval 1.6-7.0). The frequencies of common alleles were similar in patients and healthy subjects for all polymorphisms. An association between fibrinogen levels and the polymorphisms TaqI, HaeIII and BclI was found in healthy subjects, but not in the patients. It was concluded from these data that the polymorphisms TaqI, HaeIII and BclI are not major risk factors for DVT.     

New aspects in thrombotic research: complement induced switch in mast cells from a profibrinolytic to a prothrombotic phenotype

Mast cells, strategically located in the vicinity of blood vessels, are multifunctional effector cells participating in the modulation of various inflammatory and cardiovascular disease processes by actively releasing a wide variety of vasoactive mediators. These cells have also been implicated in the regulation of thrombosis and the development and progression of atherosclerosis. By expressing enzymatically active tissue type-plasminogen activator (t-PA), human mast cells (MC) might play a role in endogenous fibrinolysis and extracellular matrix remodelling--both processes that are essential in the pathogenesis of cardiovascular disorders. However, when treated with the anaphylotoxin C5a, mast cells express the PA inhibitor 1 (PAI-1) in excess over t-PA. In context with studies suggesting a role for mast cells and components of the complement system in the development of cardiovascular disease our results lead to the hypothesis that mast cells by producing t-PA in a resting state and by expressingPAI-1 when activated by C5a might participate in the modulation of the balance between proteases and protease inhibitors regulating tissue injury and repair in these disease processes. In addition, C5a might upregulate PAI-1 in mast cells to prevent its own in activation by plasmin in an autocrine or paracrine fashion.     

Chorionic gonadotropin induces dendritic cells to express a tolerogenic phenotype

The pregnancy hormone human chorionic gonadotropin (hCG) has been suggested to play an immunoregulatory role in addition to its endocrine function, thus contributing to the prevention of fetal rejection. We hypothesized that hCG is involved in the maternal-fetal immune tolerance by the regulation of dendritic cell (DC) function. Therefore, we studied the effect of hCG on DC maturation. Upon hCG treatment in combination with LPS, mouse bone marrow-derived DC (BMDC) increased the ratio of IL-10:IL-12p70, down-regulated TNF-alpha, and decreased antigen-specific T cell proliferation. Addition of hCG together with LPS and IFN-gamma blocked MHC class II up-regulation, increased IL-10 production, and decreased the antigen-specific T cell proliferation by DC. Splenic DC showed similar results. Upon hCG treatment, IDO mRNA expression and its metabolite kynurenine were increased by LPS- and IFN-gamma-stimulated DC, suggesting its involvement in the decreased T cell proliferation. To study the effect of hCG on DC differentiation from precursors, BMDC were generated in the continuous presence of hCG. Under this condition, hCG decreased cytokine production and the induction of T cell proliferation. These data are suggestive for a contribution of hCG to the maternal-fetal tolerance during pregnancy by modifying DC toward a tolerogenic phenotype.     

Incidence of deep vein thrombosis in bedridden non-surgical patients

In order to detect deep vein thrombosis (DVT), 101 patients with acute medical or infectious disorders were examined with the 125I-fibrinogen uptake test. All patients were bedridden on admission and were scanned daily from the second to the eighth day. Thirteen patients developed a positive fibrinogen uptake test. Thus, if a positive test is interpreted as DVT, the incidence of DVT was 13% in our bedridden patients. Of the patients admitted because of heart disease or pneumonia 20% had DVT, but only 4% of those admitted with other diagnoses. Other clinical "risk factors" studied, could not identify patients who developed DVT.     

缺氧诱导因子1在创伤性深静脉兔模型血栓形成及消退过程中的表达

背景：深静脉血栓形成及消退过程中的分子机制极其复杂,目前已有研究表明,缺氧与损伤因素参与了深静脉血栓形成及消退过程。 目的：观察静脉壁中缺氧诱导因子1在创伤性深静脉血栓模型兔血栓形成及消退过程中的表达变化。 方法：将日本大耳白兔采用钳夹双侧股静脉+石膏固定双下肢建立兔创伤性深静脉血栓模型。PCR及ELISA检测股静脉组织中缺氧诱导因子1 mRNA、蛋白在兔创伤性深静脉血栓建模后表达的变化。 结果与结论：模型兔创伤后24 h,经B超监测,血栓形成率为58%;血栓栓子随造模后时间延长逐渐缩小机化,创伤后第5天开始经B超监测模型组血栓形成的血管腔有断续血流信号。模型兔股静脉组织中缺氧诱导因子1在造模后1,3,5 d表达逐渐升高(P < 0.05或P < 0.01),此后7-14 d表达逐渐下降(P < 0.05或P < 0.01)。结果证实,在创伤性深静脉血栓形成过程,缺氧诱导因子1表达上调,在血栓形成后的溶解及机化再通过程中,缺氧诱导因子1表达下调。     

The ultrasound examination of the deep vein thrombosis

Ultrasonography is very useful for detection of deep vein thrombosis. The purpose of this paper is to show a method for detecting them efficiently by high resolution transducer and color Doppler system. We examined patients in the supine and prone positions. To detect the venous flow easily and differentiate thrombi from simple venous dilatation, some maneuvers are useful; one is pushing the vein area using the transducer on examination, the second is breathing overload, and the last is so-called milking. We can find throombi in the external iliac or femoral veins of patients who have symptoms of lower leg swelling, however, we need to better detect venous thrombi in the lower leg in patients with a history of pulmonary embolism. Because deep venous thrombi are increasing, the role of ultrasound will expand in the future.     

Diagnosis of deep vein thrombosis with 99Tcm-plasmin

The diagnostic efficiency of 99Tcm-plasmin test was evaluated by X-ray contrast phlebography in 110 consecutive patients with suspected deep vein thrombosis (DVT). The test was positive in 50 of 55 patients with DVT (sensitivity 91%) and negative in 18 of 55 without DVT (specificity 33%). The positive plasmin test in patients without DVT was in most cases due to another inflammatory process or a post-thrombotic state. The 99Tcm-plasmin test is a rapid and sensitive screening method for th diagnosis of DVT, but as it is based on comparison between the legs, it may be unreliable in cases of bilateral thrombosis. The low specificity makes it less valuable in patients with concomitant inflammatory disease or previous thrombosis in either leg. When the plasmin test is positive, the diagnosis of DVT should in most cases be confirmed by X-ray contrast phlebography.     

Plasma D-dimer levels in patients with deep vein thrombosis]

Hemostatic abnormalities were examined in 40 patients with deep vein thrombosis (DVT), 40 patients treated with warfarin, and 30 healthy volunteers. Plasma D-dimer levels were measured by an enzyme linked fluorescent assay (ELFA; Vidas-D-dimer) and an enzyme immunosorbent assay (ELISA; D-dimer-ELISA). Plasma levels of D-dimer, thrombin-antithrombin complex (TAT), soluble fibrin monomer (SFM) were significantly higher in patients with DVT than in the other groups. Both the sensitivity and specificity of D-dimer for diagnosis of DVT were the highest among hemostatic molecular markers examined. The most adequate cut off value of Vidas-D-dimer was 0.6 microgram/ml. Plasma levels of Vidas-D-dimer were well correlated with D-dimer ELISA, SFM and TAT. As the time of measurement for Vidas-D-dimer is less than one hour, the measurement of D-dimer using a EFLA might be useful for the diagnosis of DVT in bed side.     

亚急性创伤性深静脉血栓形成模型大鼠基质金属蛋白酶3的表达

背景：创伤性深静脉血栓发生率和死亡率呈逐年递增趋势,但其发生发展机制尚未清楚。因此有必要采用先进的基因芯片技术对其进行研究,探索其发生发展的内在规律。 目的：观察创伤性深静脉血栓形成过程中基质金属蛋白酶3的表达变化规律,探讨其在创伤性深静脉血栓形成中的作用机制。 方法：从110只SD大鼠中随机取10只大鼠作为正常组,其余100只大鼠作为实验组,采用定量击打双侧大腿+髋人字石膏固定方式,建立大鼠亚急性创伤性深静脉血栓动物模型。在伤后即刻、24,72,120和168 h时相点切取股静脉血管组织,随后抽取总RNA,采用Affymetrix 230 2.0芯片对股静脉血管组织进行基因表达检测。 结果与结论：在亚急性创伤性深静脉血栓形成演化过程中,基质金属蛋白酶3在实验组的表达随伤后时间的延长而增加,说明其参与血栓形成过程,可能是影响深静脉血栓生物学状态的主要因素之一。     

创伤性深静脉血栓形成大鼠模型Wnt信号通路的作用

背景：创伤后深静脉血栓形成具有潜在的临床危害性,可并发肺栓塞、脑栓塞。Wnt信号途径调节控制多种疾病过程,可能影响深静脉血栓的形成。 目的：探讨Wnt信号通路在创伤性深静脉血栓形成中的作用。 方法：将30只SD大鼠随机分为正常对照组和模型组。模型组根据造模后的不同生物学状态再分为高峰期血栓形成组和高峰期血栓不形成组,造模后5d无创切取股静脉血管组织,随后抽取各组大鼠总RNA,用Genechip Rat Genome 230 2.0芯片测定股静脉RNA表达,并分析Wnt信号通路基因表达变化情况。 结果与结论：与正常对照组比较,血栓形成组发现1 906个基因出现表达差异,无血栓形成组差异表达基因数目合计1 568个;与无血栓形成组比较,血栓形成组有437个出现表达差异,其中Wnt信号通路卷曲相关蛋基因、膜联结合蛋白基因、酪蛋白激酶Ⅱ基因、p53基因、蛋白磷酶2A 基因、环腺苷酸依赖性激酶同功酶基因、连接素β基因、原癌基因、fos相关抗原基因、Rac基因、钙调素依赖型蛋白激酶Ⅱ基因、钙调神经磷酸酶基因等均上调;卷曲蛋白基因、磷脂酶C基因等下调。提示Wnt信号通路可能是调控血栓的生物学状态的重要信号通路之一。     

Mallory body forming cells express the preneoplastic hepatocyte phenotype

The livers of mice fed diethyl 1,4-dihydro-2,4,6,-trimethyl-3,5-pyridinedicarboxylate (DDC) for 10 weeks formed Mallory bodies (MBs) in clusters of hepatocytes. Mice withdrawn from DDC for 9 months developed liver tumors. In the present study, the phenotype of the hepatocytes that formed MBs and tumors was characterized. Immunoperoxidase and immunofluorescent stains were done on the DDC-treated mouse livers, as well as mouse liver tumors and a human hepatocellular carcinoma that formed MBs. Antibodies to markers of hepatocellular neoplasms such as alpha-fetoprotein (AFP), ubiquitin B (UbB) fatty acid synthase (FAS) and alpha2 macroglobulin (A2m) stained the MB forming cells positive. Quantitative real-time RT-PCR assay was used to measure AFP, UbB, FAS and GCP-3 A2m mRNA levels in the livers of DDC fed mice and the DDC-induced mouse liver tumors. The FAS, UbB, GPC-3 and AFP mRNA levels were significantly increased in the MB forming liver cells. The in vitro model of MB formation was used to correlate MB formation with gene and protein expression. Primary cultures of DDC-primed hepatocytes were compared with the controls. A2m and UbB expression increased in the primary cultures of DDC-primed hepatocytes when MBs formed. Thus, the tumor markers used to identify hepatocellular carcinoma were upregulated in cells forming MBs in vivo and in vitro, suggesting that MB forming cells express preneoplastic phenotypic features.