TMEM16F mediated phosphatidylserine exposure and microparticle release on erythrocyte contribute to hypercoagulable state in hyperuricemia.

The link between hyperuricemia (HUA) and the risk of venous thromboembolism (VTE) has been well established. However, the mechanisms of thrombus generation and the effect of HUA on procoagulant activity (PCA) of erythrocytes remain unclear no matter in uremia or hyperuricemia. Here, phosphatidylserine (PS) exposure, microparticles (MPs) release, cytosolic Ca2+, TMEM16F expression, reactive oxygen species (ROS) and lipid peroxidation of erythrocyte were detected by flow cytometer. PCA was assessed by coagulation time, purified coagulation complex and fibrin production assays. The fibrin formation was observed by scanning electron microscopy (SEM). We found that PS exposure, MPs generation, TMEM16F expression and consequent PCA of erythrocyte in HUA patients significantly increased compared to those in healthy volunteers. Furthermore, high UA induced PS exposure, and MPs release of erythrocyte in concentration and time-dependent manners in vitro, which enhanced the PCA of erythrocyte and was inhibited by lactadherin, a PS inhibitor. Additionally, using SEM, we also observed compact fibrin clots with highly-branched networks and thin fibers supported by red blood cells (RBCs) and RBC-derived MPs (RMPs). Importantly, we demonstrated UA enhanced the production of ROS and lipid peroxidation and reduced the generation of glutathione (GSH) of erythrocyte, which enhanced TMEM16F activity and followed PS externalization and RMPs formation. Collectively, these results suggest that Ca2+-dependent TMEM16F activation may be responsible for UA-induced PS exposure and MPs release of RBC, which thereby contribute to the prothrombotic risk in HUA.

Hyperuricemia enhances procoagulant activity of vascular endothelial cells through TMEM16F regulated phosphatidylserine exposure and microparticle release.

The link between serum uric acid (SUA) and the risk of venous thromboembolism (VTE) is well established. Recent data suggested a causative role of UA in endothelial cells (ECs) dysfunction. However, the molecular mechanism of high UA on thrombogenesis is unknown. We investigate whether high UA induce phosphatidylserine (PS) externalization and microparticle (MP) shedding in cultured EC, and contribute to UA-induced hypercoagulable state. In the present study, we demonstrate that UA induces PS exposure and EMP release of EC in a concentration- and time-dependent manner, which enhances the procoagulant activity (PCA) of EC and inhibited over 90% by lactadherin in vitro. Furthermore, hyperuricemic rat model was used to evaluate the development of thrombi following by flow stasis in the inferior vena cava (IVC). Hyperuricemia group is more likely to form large and hard thrombi compared with control. Importantly, we found that TMEM16F expression is significantly upregulated in UA-treated EC, which is crucial for UA-induced PS exposure and MP formation. Additionally, UA increases the generation of reactive oxygen species (ROS), lipid peroxidation, and cytosolic Ca2+ concentration in EC, which might contribute to increased TMEM16F expression. Using confocal microscopy, we also observed disruption of the actin cytoskeleton, suggesting that depolymerization of actin filaments might be required for TMEM16F activation and followed by PS exposure and membrane blebbing in UA-treated EC. Our results demonstrate a thrombotic role of EC in hyperuricemia through TMEM16F-mediated PS exposure and MPs release.

Phagocytosis by endothelial cells inhibits procoagulant activity of platelets of essential thrombocythemia in vitro.

BACKGROUND: Essential thrombocythemia (ET) is characterized by thrombocytosis with increased platelet number and persistent activation. The mechanisms of thrombosis and the fate of these platelets are not clear. The aim of the present study is to explore the phagocytosis of platelets of ET patients by endothelial cells (ECs) in vitro and its relevance to the procoagulant activity (PCA). METHODS: Phosphatidylserine (PS) exposure on platelets was detected by flow cytometry. Phagocytosis of the platelets by ECs was performed using flow cytometry, confocal microscopy, and electron microscopy. The PCA of platelets was evaluated by coagulation time and purified coagulation complex assays. RESULTS: The PS exposure on platelets in ET patients is higher than that in healthy controls. The PS-exposed platelets are highly procoagulant and lactadherin reduced 80% of the PCA by blockade of PS. When cocultured, the platelets of ET patients were sequestered by ECs in a time-dependent fashion. Lactadherin enhanced phagocytosis by bridging the PS on activated platelets and the integrin αvß3 on ECs, and P-selectin played at least a partial role in this process. Furthermore, factor Xa and prothrombinase activity of PS-exposed platelets were decreased after incubation with ECs. CONCLUSION: Our results suggest that phagocytic clearance of platelets by ECs occurs in ET patients, thus representing a novel mechanism to remove activated platelets from the circulation; lactadherin and phagocytosis could cooperatively limit the thrombophilia in ET patients.

Thrombotic Role of Blood and Endothelial Cells in Uremia through Phosphatidylserine Exposure and Microparticle Release.

The mechanisms contributing to an increased risk of thrombosis in uremia are complex and require clarification. There is scant morphological evidence of membrane-dependent binding of factor Xa (FXa) and factor Va (FVa) on endothelial cells (EC) in vitro. Our objectives were to confirm that exposed phosphatidylserine (PS) on microparticle (MP), EC, and peripheral blood cell (PBC) has a prothrombotic role in uremic patients and to provide visible and morphological evidence of PS-dependent prothrombinase assembly in vitro. We found that uremic patients had more circulating MP (derived from PBC and EC) than controls. Additionally, patients had more exposed PS on their MPs and PBCs, especially in the hemodialysis group. In vitro, EC exposed more PS in uremic toxins or serum. Moreover, reconstitution experiments showed that at the early stages, PS exposure was partially reversible. Using confocal microscopy, we observed that PS-rich membranes of EC and MP provided binding sites for FVa and FXa. Further, exposure of PS in uremia resulted in increased generation of FXa, thrombin, and fibrin and significantly shortened coagulation time. Lactadherin, a protein that blocks PS, reduced 80% of procoagulant activity on PBC, EC, and MP. Our results suggest that PBC and EC in uremic milieu are easily injured or activated, which exposes PS and causes a release of MP, providing abundant procoagulant membrane surfaces and thus facilitating thrombus formation. Blocking PS binding sites could become a new therapeutic target for preventing thrombosis.

Indolic uremic solutes enhance procoagulant activity of red blood cells through phosphatidylserine exposure and microparticle release.

Increased accumulation of indolic uremic solutes in the blood of uremic patients contributes to the risk of thrombotic events. Red blood cells (RBCs), the most abundant blood cells in circulation, may be a privileged target of these solutes. However, the effect of uremic solutes indoxyl sulfate (IS) and indole-3-acetic acid (IAA) on procoagulant activity (PCA) of erythrocyte is unclear. Here, RBCs from healthy adults were treated with IS and IAA (mean and maximal concentrations reported in uremic patients). Phosphatidylserine (PS) exposure of RBCs and their microparticles (MPs) release were labeled with Alexa Fluor 488-lactadherin and detected by flow cytometer. Cytosolic Ca(2+) ([Ca(2+)]) with Fluo 3/AM was analyzed by flow cytometer. PCA was assessed by clotting time and purified coagulation complex assays. We found that PS exposure, MPs generation, and consequent PCA of RBCs at mean concentrations of IS and IAA enhanced and peaked in maximal uremic concentrations. Moreover, 128 nM lactadherin, a PS inhibitor, inhibited over 90% PCA of RBCs and RMPs. Eryptosis or damage, by indolic uremic solutes was due to, at least partially, the increase of cytosolic [Ca(2+)]. Our results suggest that RBC eryptosis in uremic solutes IS and IAA plays an important role in thrombus formation through releasing RMPs and exposing PS. Lactadherin acts as an efficient anticoagulant in this process.

[Study on distribution characteristics and potential ecological risk of soil heavy metals in the Yellow River beach region in Kaifeng City].

The distributions, soil environment status and potential ecological risk of heavy metals were studied in beach soil of returning the cropland into Yellow River beach region in Kaifeng by the Nemerows and Håkansons methods. The results showed that (1) as Among the average contents of the five heavy metals Pb, Cr, Hg, As and Cd, the highest was the average content of Cr, and the lowest was the average content of Pb and Hg. In addition to Hg, the coefficients of variation of other heavy metals were relatively small, indicating that the content of heavy metals was quite different at different sites, and to some extent, relecting that Hg, As and Pb were the major elements polluting the soil, among which, Pb pollution was the pollution with universality. There was little difference in the contents of Cr and Cd from village to village the coefficient of variation was small, and the contents were below the national standard level. (2) There was significant difference in the spatial distribution of soil heavy metal elements in the upper, the middle and lower sections of the study area. The upper section was clean, the middle section was slightly polluted, and the lower section was enriched with pollutants. (3) The distribution of heavy metals in the beach region inside and outside the levees of Yellow River was closely related to the distribution of the residential regions. In the upper section of the beach region (southwest), the population was large and the contents of heavy metals were high. The contents of heavy metals were lower in the near river zone than outside the levees of Yellow River. And the heavy metal contents in the middle and lower section were higher than those outside the levees of Yellow River, while the lower section (northwest) showed a tendency of pollution enrichment. (4) In the view of the average individual potential ecological risk index of heavy metals (E(r)i), the potential ecological risk of Hg reached intense levels, and the potential ecological risk of Pb's contribution to the integrated risk was 50.5%, which was the heavy metal with highest ecological risks. Cd and Pb had a moderate ecological risk, while As and Cr had minor ecological risk. Ecological hazards of heavy metals ranked in the ascending order of Hg > Pb > As > Cd > Cr. (5) The ecological hazard of the heavy metals was ranked in the order Hg > Cd > As > Pb > Cr. Based on the potential ecological risk level corresponding to the RI values, it was shown that there was moderate potential ecological risks of heavy metals in the Yellow River beach region in Kaifeng.

The effects of vestibular stimuli on brain cognitive processing: an ERP study.

Cognition impairment caused by space motion sickness often is a risk of spaceflight. So the dynamic changes of pattern of brain cognitive processing under varied vestibular stimuli was explored using event-related potentials. Vestibular stimulation was produced by rotary chair and varied linearly, i.e. control, constant 10 degrees/s rotation and constant 0.6 degrees/s, 0.8 degrees/s, 1.0 degrees/s, 1.2 degrees/s acceleration. Thirty-three subjects participated in the study and performed same auditory selective attention task in which the odd numbers in a randomly mixed series of odd and even numbers (go/no-go) presented acoustically in Chinese pronunciation were assigned as target signal during the rotation stimuli. The results showed that the P3 latency induced by target signal and the N1P2, P2N2 and N2P3 peak-to-peak amplitudes decreased significantly during constant 10 degrees/s rotation in contrast to control and constant angular acceleration. The P3 latency induced by target signal was shorter during constant 0.6 degrees/s and 1.2 degrees/s acceleration rotations than that during constant 0.8 degrees/s and 1.0 degrees/s acceleration rotations. It was suggested that constant angular velocity rotation had a promotion effect on brain cognitive processing. The constant angular acceleration inhibited the cognitive processing and its action had at least two different levels.

An event-related potentials study on selective attention modulated by vestibular stimulation.

OBJECTIVE: To explore the dynamic change of the late attentional selection process under linearly varied vestibular stimuli using event-related potentials (ERPs) technique. METHOD: Thirty-three subjects participated in the experiment. They were exposed to vestibular stimulation of constant angular velocity rotation (10 degrees/s) and four levels of constant angular acceleration rotation, the acceleration was 0.6 degrees/s2, 0.8 degrees/s2, 1.0 degrees/s2, 1.2 degrees/s2 respectively. The same auditory go/no-go cognitive task was done during the stimulation. The task involved verbally given Chinese digit number from two to nine with 1000 ms random interval. Subjects were asked to push the button for the odd numbers (target) and withhold to the even numbers (non-target). RESULT: Compared with control and different level of constant angular acceleration rotation, the N2 amplitude of non-target ERPs (NT-ERPs) decreased significantly over anterior-central scalp during 10 degrees/s constant rotation, but the N2 amplitude of target ERPs (T-ERPs) reduced significantly only at F4, F(Z) and T4 sites. The P3 latency of T-ERPs decreased significantly in 10 degrees/s constant rotation in contrast with control. Under four different acceleration level, the P3 latency of T-ERPs was relatively longer in 0.8 degrees/s2 and 1.0 degrees/s2, but shorter in 0.6 degrees/s2 and 1.2 degrees/s2. CONCLUSION: Constant angular velocity rotation had an activating effect on late attentional selection process. In contrast to the activation effect of constant angular velocity rotation, constant angular acceleration had an inhibition effect on the cognitive processes and this inhibition effect may have several levels.

[Study on space motion sickness before, during and after spaceflight].

Space motion sickness has long been a difficult medical problem in human space flight. Because it is a group of syndromes of the astronauts in adaptation to the new space environment and is related to a wide range of factors, the research of space motion sickness is different from that of cardiovascular and pulmonary system. Subjective methods and qualitative investigation are frequently used, and quantitative analysis is limited since objective stimuli that can be used in specific space environment are few and the samples are small. So precise localization of the problem is very difficult. In this paper space vestibular experiments and works on perceptions of space motion sickness completed by Russian and American scientists are reviewed, in the hope that it may provide some references for future space medical researches.