目的:探索在原发性膜性肾病(pMN)患者中血清磷脂酶A2受体(PLA2R)抗体水平与血栓栓塞事件的关系,探讨其作为膜性肾病血栓栓塞并发症独立预测因素的可行性.方法:临床数据集为2019年07月—2021年03月初诊于青岛大学附属医院并做过PLA2R抗体水平测定的pMN患者入院时的临床资料122例,以影像学血栓栓塞事件为分组依据分为血栓栓塞组(12例)与未发现血栓组(100例).按着血栓栓塞组与未发现血栓栓塞组1:1的比例,以临床数据为蓝本扩增数据命名为SNOTE扩增数据集(200例).先用临床数据集确定血清磷脂酶A2受体抗体水平的截断值,然后比较两组患者的临床特征.在两组数据集中使用单因素与多因素二元Logistic回归分析血栓栓塞并发症的危险因素.结果:pMN患者经过受试者工作曲线我们确定了PLA2R抗体水平的截断值为231.85 RU/ml.在临床因素的比较中收缩压、高血压史、血清白蛋白、低密度脂蛋白、血清肌酐、D二聚体异常、血清磷脂酶A2受体抗体水平方面差异有统计学意义(P<0.05).最终在多因素二元Logistic回归中高血压病史、D二聚体异常、血清磷脂酶A2受体抗体水平在两组数据集都被证明是pMN血栓栓塞并发症的独立危险因素(P<0.05).结论:血清磷脂酶A2受体抗体水平对原发性膜性肾病血栓栓塞事件有预警价值.

目的 探讨术前血清sST2浓度以及sST2联合左心房前后径(LAD)对心房颤动(房颤)射频消融术后复发的预测价值.方法 选取2020年2月至2020年10月间在西安交通大学第二附属医院心内科住院并且进行射频消融手术治疗的房颤患者84例,其中阵发性房颤57例,持续性房颤27例,男性48人,女性36人,平均年龄(64.24±10.17)岁.收集所有患者术前一般临床资料和血液学指标.采用ELISA法检测患者血清sST2浓度.心脏超声于收缩末期胸骨旁长轴切面测量LAD.所有入选的房颤患者于消融术后3、6、12月定期随访.采用单因素和多因素Cox回归分析sST2联合LAD对房颤术后复发的预测价值.结果 单因素及多因素Cox回归模型分析显示,血清sST2(HR=1.014,95％CI 0.986～1.035,P=0.035)、LAD (HR=1.271,95％CI 0.946～1.378,P=0.036)是影响房颤复发的独立危险因素.sST2联合LAD预测房颤复发的能力进一步提升,ROC曲线下面积为0.907,灵敏度0.846,特异度0.894.结论 术前检测血清sST2浓度以及sST2联合超声测量LAD可对房颤射频消融术后复发提供预测价值.

AIM:To investigate the count of circulating tissue factor-positive (TF+) procoagulant microparticles (MPs) in patients with type 1 diabetes mellitus (T1DM). METHODS:This case-control study included patients with T1DM and age and sex-matched healthy volunteers. The counts of phosphatidylserine-positive (PS+) MPs and TF+PS+MPs and the subgroups derived from different cell types were measured in the peripheral blood sample of the two groups using multicolor flow cytometric assay. We compared the counts of each MP between groups as well as the ratio of the TF+PS+MPs and PS+MPs (TF+PS+MPs/PS+MPs). RESULTS:We recruited 36 patients with T1DM and 36 matched healthy controls. Compared with healthy volunteers, PS+MPs, TF+PS+MPs and TF+PS+MPs/PS+MPs were elevated in patients with T1DM (PS+MPs: 1078.5 ± 158.08 vs 686.84 ± 122.04/μL, P <0.001; TF+PS+MPs: 202.10 ± 47.47 vs 108.33 ± 29.42/μL, P <0.001; and TF+PS+MPs/PS+MPs: 0.16 ± 0.04 vs 0.19 ± 0.05, P = 0.004), mostly derived from platelet, lymphocytes and endothelial cells. In the subgroup analysis, the counts of total and platelet TF+PS+MPs were increased in patients with diabetic retinopathy (DR) and with higher HbA1c, respectively. CONCLUSION:Circulating TF+PS+MPs and those derived from platelet, lymphocytes and endothelial cells were elevated in patients with T1DM.

The identity of vampire bat saliva anticoagulant remained elusive for almost a century. Sequencing the salivary gland genes from the vampire bat Desmodus rotundus identified Desmolaris as a novel 21.5-kDa naturally deleted (Kunitz 1-domainless) form of tissue factor pathway inhibitor. Recombinant Desmolaris was expressed in HEK293 cells and characterized as a slow, tight, and noncompetitive inhibitor of factor (F) XIa by a mechanism modulated by heparin. Desmolaris also inhibits FXa with lower affinity, independently of protein S. In addition, Desmolaris binds kallikrein and reduces bradykinin generation in plasma activated with kaolin. Truncated and mutated forms of Desmolaris determined that Arg32 in the Kunitz-1 domain is critical for protease inhibition. Moreover, Kunitz-2 and the carboxyl-terminus domains mediate interaction of Desmolaris with heparin and are required for optimal inhibition of FXIa and FXa. Notably, Desmolaris (100 μg/kg) inhibited FeCl3-induced carotid artery thrombus without impairing hemostasis. These results imply that FXIa is the primary in vivo target for Desmolaris at antithrombotic concentrations. Desmolaris also reduces the polyphosphate-induced increase in vascular permeability and collagen- and epinephrine-mediated thromboembolism in mice. Desmolaris emerges as a novel anticoagulant targeting FXIa under conditions in which the coagulation activation, particularly the contact pathway, plays a major pathological role.

Vampire bats are notorious for being the sole mammals that strictly feed on fresh blood for their survival. While their saliva has been historically associated with anticoagulants, only one antihemostatic (plasminogen activator) has been molecularly and functionally characterized. Here, RNAs from both principal and accessory submaxillary (submandibular) salivary glands of Desmodus rotundus were extracted, and similar to 200 million reads were sequenced by Illumina. The principal gland was enriched with plasminogen activators with fibrinolytic properties, members of lipocalin and secretoglobin families, which bind prohemostatic prostaglandins, and endonucleases, which cleave neutrophil-derived procoagulant NETs. Anticoagulant (tissue factor pathway inhibitor, TFPI), vasodilators (PACAP and C-natriuretic peptide), and metalloproteases (ADAMTS-1) were also abundantly expressed. Members of the TSG-6 (anti-inflammatory), antigen 5/CRISP, and CCL28-like (antimicrobial) protein families were also sequenced. Apyrases (which remove platelet agonist ADP), phosphatases (which degrade procoagulant polyphosphates), and sphingomyelinase were found at lower transcriptional levels. Accessory glands were enriched with antimicrobials (lysozyme, defensin, lactotransferrin) and protease inhibitors (TIL-domain, cystatin, Kazal). Mucins, heme-oxygenase, and IgG chains were present in both glands. Proteome analysis by nano LC-MS/MS confirmed that several transcripts are expressed in the glands. The database presented herein is accessible online at http://exon.niaid.nih.gov/transcriptome/D_rotundus/Supplemental-web.xlsx. These results reveal that bat saliva emerges as a novel source of modulators of vascular biology.Biological significanceVampire bat saliva emerges as a novel source of antihemostatics which modulate several aspects of vascular biology. Published by Elsevier B.V.

The development of the Cre recombinase-controlled (Cre/LoxP) technique allows the manipulation of specific tumorigenic genes, temporarily and spatially. Our original intention of this study was to investigate the role of Kras and p53 in the development of urinary bladder cancer. First, to validate the effect of intravesical delivery on Cre recombination (Adeno-Cre), we examined activity and expression of β-galactosidase in the bladder of control ROSA transgenic mice. The results confirmed specific recombination as evidenced by β-galactosidase activity in the bladder urothelium of these mice. Then, we administered the same adenovirus into the bladder of double transgenic KrasLSLG12D/+. p53fl/fl mice. The virus solution was held in place by a distal urethral retention suture for 2 hours. To our surprise, there was a rapid development of a spindle-cell tumor with sarcoma characteristics near the suture site, within the pelvic area but outside the urinary track. Since we did not see any detectable β-galactosidase in the area outside of the bladder in the validating (control) experiment, we interpreted that this sarcoma formation was likely due to transduction by Adeno-Cre in the soft tissue of the suture site. To avoid the loss of skin integrity associated with the retention suture, we transitioned to an alternative technique without suture to retain the Adeno-Cre into the bladder cavity. Interestingly, although multiple Adeno-Cre treatments were applied, only urothelial hyperplasia but not carcinogenesis was observed in the subsequent experiments of up to 6 months. In conclusion, we observed that the simultaneous inactivation of p53 and activation of Kras induces quick formation of spindle-cell sarcoma in the soft tissues adjacent to the bladder but slow formation of urothelial hyperplasia inside the bladder. These results strongly suggest that the effect of oncogene regulation to produce either hyperplasia or carcinogenesis greatly depends on the tissue type.

Atrial fibrillation (AF) and venous thromboembolism (VTE) are the two most common medical conditions managed with anti-coagulation therapy. Not all the patients with decreased mobility or AF have a similar risk for thromboembolism. The risk factors for venous thromboembolism and thromboembolism associated with AF are described in various studies. Considering that the two conditions have similar pathophysiologic basis of clot formation, one could imply that the risk factors for the occurrence of thrombosis could be similar. The present review focuses on the similarities and differences in the clinical risk factors of VTE and AF related thromboembolism. We will also be discussing the role of CHADS2-VASc scoring system in the risk assessment of VTE.

SummarySalivary glands from haematophagous animals express a notable diversity of negative modulators of platelet function. Triplatin is an inhibitor of collagen-induced platelet aggregation which has been described as an antagonist of glycoprotein VI (GPVI). Because triplatin displays sequence homology to members of the lipocalin family of proteins, we investigated whether triplatin mechanism of action could be explained by interaction with pro-haemostatic prostaglandins. Our results demonstrate that triplatin inhibits platelet aggregation induced by low doses of collagen, thromboxane A2 (TXA2) mimetic (U46619), and arachidonic acid (AA). On the other hand, it does not inhibit platelet aggregation by convulxin, PMA, or low-dose ADP. Isothermal titration calorimetry (ITC) revealed that triplatin binds AA, cTXA2, TXB2, U46619 or prostaglandin (PG)H2 mimetic (U51605). Consistent with its ligand specificity, triplatin induces relaxation of rat aorta contracted with U46619. Triplatin also interacts with PGF2a and PGJ2, but not with leukotrienes, AA or biogenic amines. Surface plasmon resonance experiments failed to demonstrate interaction of triplatin with GPVI; it also did to inhibit platelet adhesion to fibrillar or soluble collagen. Because triplatin displays sequence similarity to apolipoprotein D (ApoD) – a lipocalin associated with high-density lipoprotein, ApoD was tested as a putative TXA2-binding molecule. ITC failed to demonstrate binding of ApoD to all prostanoids described above, or to AA. Furthermore, ApoD was devoid of inhibitory properties towards platelets activation by AA, collagen, or U46619. In conclusion, triplatin mechanism of action has been elucidated without ambiguity as a novel TXA2- and PGF2α- binding protein. It conceivably blocks platelet aggregation and vasoconstriction, thus contributing to successful blood feeding at the vector-host interface.

Objective It is to study the electrophysiological characteristics of cardiocyte from pulmonary vein cardiac muscle(PVC) of rabbit applying intracellular recording technique of routine glass microelectrode,to observe the electrical conduction between PVC and left atrium cardiocyte(LAC) and to approach the mechanism of paroxysmal atrial fibrillation originated from pulmonary vein.Methods Multiple cardiocytes preparations from pulmonary vein and left atrium of 50 healthy adult rabbits were obtained.The routine intracellular microelectrode technique was applied to record action potential(AP) of PVC and LAC,and a comparison of action potential duration(APD20,APD50,APD90) between them was made.Different frequency of pulse stimulation was given at PVC terminal and LAC terminal respectively and the generated information of AP at the other terminal was observed.Two mmol/L Caesium chloride(CsCl) was added into the superfusate,and the occurred probability of early after depolarization(EAD) in the two parts was observed.Results APD20,APD50,APD90 of PVC were all longer than those of LAC(APD20 23.09±5.44 vs 19.50±1.66,APD50 52.40±7.93 vs 28.90±4.36,APD90 123.35±8.26 vs 69.65±9.44) and the difference was significant(all P<0.05).At determinate extent of stimulation frequency,the AP signals from PVC could be conducted freely to LAC.On the contrary,the conduction signals from LAC could seldom reach PVC(100% vs 10%,P<0.05).EAD was more easily induced in PVC after drug perfusion added(90% vs 15%,P<0.05).Conclusion The conduction of AP between PVC and LAC was unequal,which may be the basis of reentry formation.CsCl,as a K+ channel blockers,can lengthen repolarization process and create the condition of the occurrence of after depolarization.The long APD characteristics of PVC have the proneness of early after depolarization occurred and the excitation is easy conduct into left atrium,which maybe promote the occurred basis of the atrial arrhythmia.