优秀女子赛艇运动员比赛前后血乳酸、血尿素及血清肌酸激酶跟踪测试

目的:跟踪测试优秀女子赛艇运动员比赛前后血乳酸、血尿素及血清肌酸激酶的变化,探讨女子赛艇运动员机体能量物质代谢和赛后疲劳恢复的特点。方法:选取2005-07/09湖北省水上运动中心优秀女子运动员14名,其中国家健将级运动员7名,国家一级运动员7名;平均年龄(22±4)岁,训练年限(4.1±1.4)年,身高(174.2±3.1)cm,体质量(68.6±4.7)kg。14名运动员分别于2000m划艇比赛当日晨空腹、赛前15min、赛后5min,1h及24h抽取指尖血60μL,进行血乳酸、血尿素、血清肌酸激酶检测。结果:14名运动员全部进入结果分析。①比赛前后运动员血乳酸水平的变化:运动员空腹血乳酸水平为(1.22±0.45)mmol/L,赛前15min为(2.89±0.49)mmol/L。赛后5min血乳酸浓度高达(11.51±1.72)mmol/L,与空腹水平比较差异有显著性意义(t=3.077,P<0.01)。赛后24h血乳酸浓度显著下降至(1.76±0.24)mmol/L,与空腹水平基本接近(t=0.027,P>0.05)。②比赛前后运动员血尿素水平的变化:与空腹血尿素水平比较,运动员赛后5min血尿素浓度明显升高[(5.45±0.47),(6.13±1.00)mmol/L;t=2.416,P<0.05]。赛后24h血尿素浓度下降至(5.94±0.85)mmol/L,仍高于空腹水平(t=2.682,P<0.05)。③比赛前后运动员血清肌酸激酶活性的变化:与赛前比较,运动员赛后5min血清肌酸激酶活性明显升高[(3.38±1.58),(6.13±3.25)nkat/L;t=4.968,P<0.01]。赛后1h血清肌酸激酶活性开始下降,至赛后24h与赛前基本相似(t=1.537,P>0.05)。结论:①赛后5min血乳酸、血尿素、血清肌酸激酶活性显著高于赛前,赛后1h血乳酸消除迅速,但仍未恢复到正常水平。提示赛艇是一种以糖酵解系统为主、无氧 有氧代谢混合型供能的运动项目。②赛艇比赛使酸性产物生成增多,血乳酸、血尿素、血清肌酸激酶可作为运动强度和机能恢复的指标。③赛艇比赛后至少24h内,机体处于蛋白质降解增强状态,建议恢复期增加饮食糖和蛋白质摄入量,以促进合成代谢,加快功能恢复过程。     

The genome of Aiptasia,a sea anemone model for coral symbiosis

The most diverse marine ecosystems, coral reefs, depend upon a functional symbiosis between a cnidarian animal host (the coral) and intracellular photosynthetic dinoflagellate algae. The molecular and cellular mechanisms underlying this endosymbiosis are not well understood, in part because of the difficulties of experimental work with corals. The small sea anemone Aiptasia provides a tractable laboratory model for investigating these mechanisms. Here we report on the assembly and analysis of the Aiptasia genome, which will provide a foundation for future studies and has revealed several features that may be key to understanding the evolution and function of the endosymbiosis. These features include genomic rearrangements and taxonomically restricted genes that may be functionally related to the symbiosis, aspects of host dependence on alga-derived nutrients, a novel and expanded cnidarian-specific family of putative pattern-recognition receptors that might be involved in the animal–algal interactions, and extensive lineage-specific horizontal gene transfer. Extensive integration of genes of prokaryotic origin, including genes for antimicrobial peptides, presumably reflects an intimate association of the animal–algal pair also with its prokaryotic microbiome.Coral reefs form marine-biodiversity hotspots that are of enormous ecological, economic, and aesthetic importance. Coral growth and reef deposition are based energetically on the endosymbiosis between the cnidarian animal hosts and photosynthetic dinoflagellate algae of the genus Symbiodinium, which live in vesicles within the gastrodermal (gut) cells of the animal and typically supply ≥90% of its total energy, while the host provides the algae with a sheltered environment and the inorganic nutrients needed for photosynthesis and growth (1). This tight metabolic coupling allows the holobiont (i.e., the animal host and its microbial symbionts) to thrive in nutrient-poor waters. Although the ecology of coral reefs has been studied intensively, the molecular and cellular mechanisms underlying the critical endosymbiosis remain poorly understood (2). As coral reefs face an ongoing and increasing threat from anthropogenic environmental change (3), new insights into these mechanisms are of critical importance to understanding the resilience and adaptability of coral reefs and thus to the planning of conservation strategies (4).Aiptasia is a globally distributed sea anemone that harbors endosymbiotic Symbiodinium like its Class Anthozoa relatives the stony corals (Fig. 1 and SI Appendix, Fig. S1A) (4, 5). Aiptasia has a range of polyp sizes convenient for experimentation and is easily grown in laboratory culture, where it reproduces both asexually (so that large clonal populations can be obtained) and sexually (allowing experiments on larvae and potentially genetic studies), and it can be maintained indefinitely in an aposymbiotic (dinoflagellate-free) state and reinfected by a variety of Symbiodinium strains (6, 7). These characteristics make Aiptasia a highly attractive model system for studies of the molecular and cellular basis of the cnidarian–dinoflagellate endosymbiosis (2, 4). To provide a solid platform for research on Aiptasia, we have sequenced and analyzed its genome. The results have already provided important insights into several aspects of the evolution and function of the symbiotic system.Open in a separate windowFig. 1.Phylogenetic position and different symbiotic states of Aiptasia. (A) Partial phylogenetic tree (see SI Appendix, SI Materials and Methods and Fig. S1A for details) shows Aiptasia grouped with other anthozoans among the cnidarians. Numbers on nodes denote bootstrap values. (B–D) An aposymbiotic Aiptasia polyp (B) and symbiotic polyps viewed under white light (C) or by fluorescence microscopy to visualize the red chlorophyll autofluorescence of the endosymbiotic Symbiodinium algae (D).     

Preparation of factor IX deficient human plasma by immunoaffinity chromatography using a monoclonal antibody

A murine hybridoma clone is described that grows continuously in culture and produces a monoclonal antibody we have called Royal Free Monoclonal Antibody to factor IX No. 1 (RFF-IX/1). This has high affinity for a coagulation site on factor IX. RFF-IX/1 immobilised on sepharose can be used to deplete factor IX from normal human plasma. This immunoaffinity depleted plasma is indistinguishable from severe Christmas disease plasma and can be used as the substrate in a one stage coagulation assay for factor IX. The affinity column has high capacity and can be regenerated so that large scale production from normal plasma of factor IX deficient plasma as a diagnostic reagent is now feasible.     

High-molecular-weight kininogen is exclusively membrane bound on endothelial cells to influence activation of vascular endothelium

An important biologic function of high-molecular-weight kininogen (HK) is to deliver bradykinin (BK) to its cellular receptors. Internalization and degradation of HK may provide a mechanism by which endothelial cells modulate the production of BK and control its activities. Therefore, we investigated the binding and subsequent distribution of biotinylated-HK (biotin-HK) associated with human umbilical vein endothelial cells (HUVEC). HUVEC bound 3 to 4 times more HK and with greater avidity at 1 to 3 hours at 37 degrees C than at 4 degrees C (Bmax = 1.0 +/- 0.02 x 10(7) molecules/cell, kd = 7 +/- 3 nmol/L v Bmax = 2.6 +/- 0.2 x 10(6) molecules/cell, kd = 46 +/- 8 nmol/L). However, there was no evidence that the difference was caused by internalization of HK at the higher temperature. First, the same amount of biotin-HK was associated with nonpermeabilized and permeabilized HUVEC using buffers containing 20 to 50 mumol/L zinc ion in the absence or presence of 2 mmol/L calcium ion. Second, binding of biotin-HK to HUVEC was approximately 92% reversible at 1 hour when the cells were maintained at both 37 degrees C and 4 degrees C. Third, neither chloroquine nor primaquine altered the amount of biotin-HK bound to HUVEC. Fourth, biotin-HK bound to HUVEC was almost completely removed by pronase. Fifth, the nonpermeable dye, crystal violet, almost completely quenched the fluorescence signal emitted by HUVEC-associated fluorescein isothiocyanate (FITC) HK. Finally, the localization of HUVEC-bound FITC-HK was restricted to the membrane as shown by laser scanning confocal microscopy. The expression of HK binding sites had an absolute requirement for metabolic energy, but was not dependent on new protein synthesis. Membrane-bound HK contributed to the anticoagulant nature of endothelial cells by blocking human alpha-thrombin binding and its resultant induction of prostacyclin formation. These studies indicate that HK is not internalized by HUVEC, but remains primarily on cell surfaces to be accessible for BK liberation and to modulate the binding and actions of alpha-thrombin.     

High molecular weight kininogen: localization in the unstimulated and activated platelet and activation by a platelet calpain(s)

High mol wt kininogen (HMWK), the major cofactor-substrate of the contact phase of coagulation, is contained within and secreted by platelets. Studies have been performed to localize platelet HMWK in both the unstimulated and activated platelet and to ascertain the effect of platelet enzymes on HMWK itself. On platelet subcellular fractionation, platelet HMWK was localized to alpha-granules, and platelets from a patient with a deficiency of these granules (gray platelet syndrome) had 28% normal platelet HMWK. Platelet HMWK, in addition to being secreted from the platelet, was also localized to the surface of the platelet when activated. Using a competitive enzyme- linked immunosorbent assay for HMWK as an indirect antibody consumption assay, the external membrane of thrombin-activated platelets as well as the releasate from these stimulated platelets had 17 ng HMWK antigen/10(8) platelets available, whereas unstimulated platelets and their supernatant had only 4.9 and 4.2 ng HMWK/10(8) platelets present, respectively. The anti-HMWK antibody consumption by activated normal platelets was specific for membrane-expressed platelet HMWK, since activated platelets from a patient with total kininogen deficiency did not adsorb the anti-HMWK antibody. Enzymes in the cytosolic fraction of platelets cleaved 125I-HMWK (mol wt 120,000) into a mol wt 100,000 polypeptide as well as smaller products at mol wt 74,000, mol wt 62,000, mol wt 47,000, and a few components below mol wt 45,000. No cleavage products were observed when DFP and leupeptin were present. The cleavage of HMWK was specifically prevented by inhibitors of calcium-activated cysteine proteases (leupeptin, N-ethylmaleimide, iodoacetamide, and EDTA) but not by inhibitors of serine proteases (DFP, benzamidine, soybean trypsin inhibitor, or aprotinin). Platelet cytosol increased the coagulant activity of exogenous purified HMWK with maximum HMWK coagulant activity (35-fold) occurring within ten minutes of exposure to platelet cytosol. Treatment of platelet cytosol with leupeptin prevented the increase in the coagulant activity of exogenous HMWK. These studies indicate that activated platelets express platelet HMWK on their external membrane and platelet enzymes can cleave and increase the coagulant activity of exogenous HMWK.     

Sulfide silver amplification of ferritin iron cores in blood and bone marrow cells

Summary A short exposure of cell suspensions to gaseous hydrogen sulfide, appropriate fixations, and subsequent physical development of silver shells around sulfidated insoluble metals were used to amplify ferritin iron cores in blood and bone marrow cells. The methods described are suitable for both light microscopy and transmission electron microscopy. These techniques made it possible to visualize Prussian Blue stainable ferritin and haemosiderin, as well as a large variety of isoferritin iron and other smaller particles beyond the sensitivity of Prussian Blue staining. Admixtures of sulfidatible zinc and traces of other heavy metals had to be taken into consideration. For further research, adaptations of sulfide silver staining to microphysical analyses were developed. However, conventional energy dispersive X-ray analysis was not sensitive enough to signalize the presence of Fe in sulfide silver amplified iron cores of a single or a few ferritin molecule(s). Proton-induced X-ray emission was used to measure Fe and Zn down to 1 fg/single cell in unstained or sulfide silver stained smears on thin foils. However, multielement analysis of homogeneous cell concentrates was much easier to perform and far more sensitive. In advanced iron overload, highly increased sulfide silver staining was found in peripheral blood cells including lymphocytes, monocytes, eosinophils, basophils, and — in extreme cases — also in neutrophils and platelets.     

The effect of pharmacological agents on the rate of lymph outflow and on blood and lymph toxicity in acute myocardial infarct]

In experiments on rabbits it was found that acute myocardial infarction (AMI) was associated with a disorder of the drainage function the lymphatic system and a drastic increase of the lymph toxicity. The administration of propranolol (obsidan), amiodaron (cordaron), lidocaine (xycaine), trimecaine (mesocaine), nitroglycerin, panangin and heparin exerted the pronounced stimulating action on the lymph outflow rate. Strophanthin-K, corglycon and digo in possessed the moderate lymphogenic effect. Novocainamide (procainamide), verapamil (isoptine) and panangin exerted no influence on the lymph outflow rate. The administration of the above mentioned drugs in the initial period of AMI sharply increased the toxic properties of the lymph; subsequently the lymph toxicity gradually decreased and was less than in control (in AMI treated with drugs).