ABO blood types: influence on infarct size, procedural characteristics and prognosis

Introduction

Patients with non-O blood groups have higher plasma von Willebrand factor (vWF) levels than those with type O. vWF mediates platelet adhesion, aggregation and thrombosis. These considerations likely explain the prior observations that non-O patients have higher rates of arterial and venous thromboembolic events. However, the effect of blood group status on size of MI, procedural findings and outcomes after PCI for MI have not been reported.

Methods

We analyzed 1198 patients who underwent percutaneous coronary intervention for acute myocardial infarction between 10/03 and 8/06, and who had ABO blood group status and clinical follow-up.

Results and conclusions

Patients with O blood type were slightly older (62 ± 13 vs. 60 ± 13years; p = 0.017) had a higher prevalence of hypercholesterolemia (67% vs. 58%; p = 0.002), and had a higher burden of atherosclerosis with more vascular disease (17% vs. 13%; p = 0.017) and higher prevalence of previous PCI (22% vs. 17%; p = 0.025). Non-O blood group patients had larger infarcts as measured by median peak troponin (33 vs. 24; p = 0.037), total CK (721 vs. 532; p = 0.012) and CK-MB (101 vs. 68; p = 0.010). At PCI, non-O patients had increased visible thrombus and reduced TIMI flow pre-procedure. However, there were no differences in procedural success, in-hospital blood transfusion or occurrence of MACE at 1year follow-up. Our data demonstrate that non-O compared to O blood groups patients have higher thrombus burden despite less extensive atherosclerosis. Nevertheless, outcomes at 1year were similar.     

高效液相色谱法测定血清中双氯芬酸浓度及在人体的药代动力学

建立了人血清中双氯芬酸的反相高效液相色谱测定方法。该法操作简便、精密度好、方法回收率为９８．２～１０２．５％，日内、日间ＲＳＤ为１．２９～４。５２％，血药浓度在０．２～１０μｇ／ｍｌ范围内呈线性关系，相关系数０；９９９９，当信噪比为２时，最低检测限为１０ｎｇ／ｍｌ。健康志愿者口服１００ｍｇ双氯芬酸肠溶衣片，药代动力学过程符合一室开放模型，消除半衰期为２．１５ｈ。     

3－硝基－1，2，4－三唑类乏氧细胞放射增敏剂的构效关系

为了寻找毒性低、增敏作用强的乏氧细胞放射增敏剂，设计并合成了一系列５－溴－，５－甲基－，和５－未取代的３－硝基－１，２，４－三唑－１－乙酰胺类化合物，用ＨｅＬａＳ３细胞进行了体外试验。结果表明５－溴取代衍生物的增敏作用强于相应的５－甲基－或５－未取代的硝基三唑衍生物，但是它们的毒性亦增大。修饰１位乙酰胺侧链也可以改变化合物的增敏作用和亲脂性。在所测定的化合物中ＴＡ－１０１［２－（３－硝基－１－三唑基）乙酰胺］由于有高的增敏作用和低亲脂性，可能是一个有希望的放射增敏剂。     

促红细胞生成素对实验性肾性贫血的作用

促红细胞生成素(erythropoietin,EPO)是由肾细胞分泌的一种糖蛋白激素。从人胚肾细胞中诱导,经生物化学方法分离、提纯得到此品。本试验用5/6肾切除的方法造成大鼠慢性肾衰性(CRF)贫血,研究不同剂量EPO对CRF贫血的作用。结果表明EPO有显著的促进红细胞生成,改善CRF贫血状态,使其接近或达到正常水平,最佳剂量为1000 U/kg,并可预防实验性贫血,对正常鼠未见明显作用。     

原代培养鼠胚脊髓运动神经元活力状态与肌萎灵注射液的干预作用

目的:从细胞水平,观察扶元起萎,养荣生肌的中药制剂肌萎灵注射液对原代培养鼠胚脊髓运动神经元的保护作用。方法:实验于2004-03/2005-03在解放军第三军医大学完成。实验分组:清洁级SD雌性孕鼠,孕期10～14d。应用密度梯度离心法分离鼠胚脊髓运动神经元进行原代培养,运动神经元培养72h后,按培养板及孔分为4.5μg/L肌萎灵组(肌萎灵注射液,含生药0.9g/mL)、9μg/L肌萎灵组、45μg/L肌萎灵组、力如太组(力如太R利鲁唑片,应用时经0.9%NaCl-0.01NHCl溶解)和对照组。实验处理:各组分别加入用新鲜培养基稀释为0.5%(终浓度4.5μg/L)、1%(终浓度9μg/L)、5%(终浓度45μg/L)的肌萎灵注射液,力如太组加入利鲁唑(终浓度10μmo/L),对照组加入等量新鲜培养基。实验评估:①共同培养3d用四甲基偶氮唑盐法观察其对细胞活力的影响,以吸光度表示。②采用NF-200免疫组织化学染色并进行图像分析,测定神经突起主干长度。结果:①培养的运动神经元活力状态比较:4.5,9,45μg/L肌萎灵组培养运动神经元活力显著增强,与对照组相比,差异有显著性意义(0.317±0.054,0.396±0.087,0.329±0.097,0.230±0.130,P<0.05)。力如太组细胞生长与对照组相比无显著差异(0.266±0.141,0.230±0.130,P>0.05)。②脊髓运动神经元突起生长情况结果:4.5μg/L肌萎灵组和9μg/L肌萎灵组可促进其生长,与对照组相比,差异有显著性意义[(315.96±32.32),(373.46±80.24),(159.71±48.95)μm,P<0.05]。结论:肌萎灵注射液可增强运动神经元活力,促进脊髓运动神经元突起的生长。     

Estrogen mediates Aurora-A overexpression, centrosome amplification, chromosomal instability, and breast cancer in female ACI rats

Estrogens play a crucial role in the causation and development of sporadic human breast cancer (BC). Chromosomal instability (CIN) is a defining trait of early human ductal carcinoma in situ (DCIS) and is believed to precipitate breast oncogenesis. We reported earlier that 100% of female ACI (August/Copenhagen/Irish) rats treated with essentially physiological serum levels of 17beta-estradiol lead to mammary gland tumors with histopathologic, cellular, molecular, and ploidy changes remarkably similar to those seen in human DCIS and invasive sporadic ductal BC. Aurora-A (Aur-A), a centrosome kinase, and centrosome amplification have been implicated in the origin of aneuploidy via CIN. After 4 mo of estradiol treatment, levels of Aur-A and centrosomal proteins, gamma-tubulin and centrin, rose significantly in female ACI rat mammary glands and remained elevated in mammary tumors at 5-6 mo of estrogen treatment. Centrosome amplification was initially detected at 3 mo of treatment in focal dysplasias, before DCIS. At 5-6 mo, 90% of the mammary tumor centrosomes were amplified. Comparative genomic hybridization revealed nonrandom amplified chromosome regions in seven chromosomes with a frequency of 55-82% in 11 primary tumors each from individual rats. Thus, we report that estrogen is causally linked via estrogen receptor alpha to Aur-A overexpression, centrosome amplification, CIN, and aneuploidy leading to BC in susceptible mammary gland cells.     

SLO3 auxiliary subunit LRRC52 controls gating of sperm KSPER currents and is critical for normal fertility

Following entry into the female reproductive tract, mammalian sperm undergo a maturation process termed capacitation that results in competence to fertilize ova. Associated with capacitation is an increase in membrane conductance to both Ca²⁺ and K⁺, leading to an elevation in cytosolic Ca²⁺ critical for activation of hyperactivated swimming motility. In mice, the Ca²⁺ conductance (alkalization-activated Ca²⁺-permeable sperm channel, CATSPER) arises from an ensemble of CATSPER subunits, whereas the K⁺ conductance (sperm pH-regulated K⁺ current, KSPER) arises from a pore-forming ion channel subunit encoded by the slo3 gene (SLO3) subunit. In the mouse, both CATSPER and KSPER are activated by cytosolic alkalization and a concerted activation of CATSPER and KSPER is likely a common facet of capacitation-associated increases in Ca²⁺ and K⁺ conductance among various mammalian species. The properties of heterologously expressed mouse SLO3 channels differ from native mouse KSPER current. Recently, a potential KSPER auxiliary subunit, leucine-rich-repeat-containing protein 52 (LRRC52), was identified in mouse sperm and shown to shift gating of SLO3 to be more equivalent to native KSPER. Here, we show that genetic KO of LRRC52 results in mice with severely impaired fertility. Activation of KSPER current in sperm lacking LRRC52 requires more positive voltages and higher pH than for WT KSPER. These results establish a critical role of LRRC52 in KSPER channels and demonstrate that loss of a non-pore-forming auxiliary subunit results in severe fertility impairment. Furthermore, through analysis of several genotypes that influence KSPER current properties we show that in vitro fertilization competence correlates with the net KSPER conductance available for activation under physiological conditions.Upon entry into the female reproductive tract, mammalian sperm undergo a sequence of maturational steps, collectively termed capacitation, to become competent to fertilize an egg (1, 2). Two important components of this process, thought to be shared among mammalian species, are cytosolic alkalization (3–5) and then an associated increase in cytosolic Ca²⁺ (6, 7). These events are coupled with changes in ionic fluxes in sperm membrane. Over the past 10 y, the application of patch-clamp recording to individual sperm (8) has allowed identification of ionic currents that respond to alkalization and/or Ca²⁺ (9–12). In mouse sperm, alkalization leads to activation of two sperm-specific channels, the Ca²⁺-permeable CATSPER channel (8–10, 13) and the K⁺-permeable KSPER K⁺ channel (14, 15). KSPER and CATSPER currents are also present in human sperm (16–18), although intriguingly there seem to be differences in regulation of each channel type between mice and humans (11, 12, 17).Despite the species-specific differences in the details of their regulation, KSPER and CATSPER are of central importance in sperm function and fertility in both humans and mice. In mouse sperm, KSPER and CATSPER together account for all patch-clamp measurable cation current activated by voltage and alkalization (19) and are thought to act in concert to mediate the changes in membrane cation conductance and Ca²⁺ influx that occur during the onset of capacitation (14, 20). The critical role of both KSPER and CATSPER in the mouse has been established by demonstration that genetic KO of the pore-forming subunits of each channel [KSPER (15, 21) and CATSPER (22–26)] results in infertility and the CATSPER auxiliary δ subunit is also required for fertility (27).For mouse KSPER, the pore-forming subunit is termed SLO3, encoded by the kcnu1 (slo3) gene (28). SLO3, which is exclusively expressed in testis (15, 28), is a homolog of the Ca²⁺- and voltage-activated, large-conductance (BK)-type K⁺ channel (29) and heterologously expressed SLO3 results in voltage- and alkalization-activated K⁺ currents (28). However, heterologously expressed mouse SLO3 channels differ from mouse KSPER current in important ways. Specifically, whereas pH 7 substantially activates KSPER in mouse sperm at membrane potentials (V_m) between −10 and −50 mV, activation of SLO3 channels at pH 7 is scarcely observed even at an activation potential of +100 mV (30, 31). This discrepancy raised the possibility that an additional regulatory partner of SLO3 may be present in mouse sperm. Guided by the discovery of a new type of auxiliary subunit of the BK channel (32), we recently showed that a related subunit, LRRC52 (leucine-rich-repeat-containing protein 52) is selectively expressed in sperm (33). When LRRC52 is coexpressed with SLO3, the gating range of the resulting channels at a given pH is more like that of KSPER in mouse sperm. LRRC52 protein has also been shown to be present in human sperm and, when coexpressed with human SLO3, LRRC52 results in currents with properties similar to those of human KSPER (12). To evaluate the importance of the LRRC52 subunit, we have now generated lrrc52^−/− (LRRC52 KO) mice. LRRC52 KO mice have a severe fertility deficit that is associated with a shift to more positive potentials in the KSPER current activation range in the LRRC52 KO sperm. Furthermore, measurement of the net KSPER current in sperm from two other genotypes, slo3^+/− and a slo3-eGFP, suggests that sperm reproductive capacity strongly depends on the KSPER conductance available over physiological potentials. These results confirm that LRRC52 is a critical component of the KSPER channel complex and is essential for male fertility.     

Centrosome hypertrophy in human breast tumors: Implications for genomic stability and cell polarity

The centrosome plays an important role in maintenance of cell polarity and in progression through the cell cycle by determining the number, polarity, and organization of interphase and mitotic microtubules. By examining a set of 35 high grade human breast tumors, we show that centrosomes of adenocarcinoma cells generally display abnormal structure, aberrant protein phosphorylation, and increased microtubule nucleating capacity in comparison to centrosomes of normal breast epithelial and stromal tissues. These structural and functional centrosome defects have important implications for understanding the mechanisms by which genomic instability and loss of cell polarity develop in solid tumors.