脉血康胶囊联合瑞舒伐他汀对冠脉临界病变患者冠脉病变程度的影响

目的研究脉血康胶囊联合瑞舒伐他汀对冠脉临界病变患者钙库操纵性钙通道蛋白(SOCC)基质交联分子1(STIM1)、钙释放激活钙通道蛋白Orai1及瞬时受体电位通道1(TRPC1)蛋白水平和冠脉病变严重程度的影响。方法将2015年1月—2016年12月北京安贞医院经定量冠脉造影检查确诊冠脉临界病变(BCL)住院患者160例设为临界病变组(BCL组),150例无冠脉病变的受试者为对照组(简称CTR组)。BCL组随机分为常规治疗组(简称RTT组,80例)和联合治疗组(简称CBT组,80例)。RTT组给予生活方式改善、阿司匹林、AECI、β受体阻滞剂及瑞舒伐他汀10 mg等药物。CBT组在常规药物治疗基础上,每日加用瑞舒伐他汀15 mg和脉血康胶囊3.0 g,连续治疗12个月。检测血小板STIM1、Orai1、TRPC1、高敏C-反应蛋白(hs-CRP)水平及TC、LDL-C、HDL-C、TG浓度。评价BCL组平均狭窄程度(MPS)。结果BCL组130例受试者复查冠脉造影。与本组治疗前比较,治疗后CBT组和RTT组血小板STIM1、Orai1、TRPC1水平均降低(P<0.05);CBT组冠脉临界病变的MPS明显减低(59.74±9.72vs.38.92±13.84,P<0.05),RTT组MPS有所下降(58.96±8.67vs.55.43±10.03),但差异无统计学意义(P>0.05)。与RTT组比较,治疗后CBT组血小板STIM1、Orai1、TRPC1、TC、LDL-C、TG、和hs-CRP明显降低(P<0.05,P<0.01),且CBT组MPS差值大于RTT组(P<0.01)。结论脉血康胶囊3.0 g和瑞舒伐他汀15 mg每日联合应用可降低SOCC水平,其可能是冠脉临界病变治疗的优化选择。     

真菌性、细菌性和病毒性肺炎的若干凝血指标变化分析

探讨真菌性、细菌性和病毒性肺炎的若干凝血指标差异性研究。方法 回顾性分析2017 年7 月～2019 年11 月本院肺炎患者128 例与同期门诊健康体检者50 例，分析患者血浆纤维蛋白原（FIB）、凝血酶原时间（PT）和活化部分凝血酶时间（APTT）的水平。结果 三类肺炎组与对照组在FIB、PT 和APTT 水平比较，差异均有统计学意义（P＜0.01）；FIB、PT 水平与病原感染呈正相关，APTT 与病原感染呈负相关。结论FIB、PT、APTT 可以间接提示肺炎感染病原体类型，具有临床意义。     

Successful treatment of linezolid-induced severe lactic acidosis with continuous venovenous hemodiafiltration: A case report

Linezolid is an oxazolidinone antibiotic. Linezolid-associated lactic acidosis has been reported in 6.8% of linezolid-treated patients. Lactic acidosis is associated with poor clinical outcomes, with high blood lactate levels resulting in organ dysfunction and mortality. This case report describes the development of lactic acidosis in a 64-year-old Chinese woman who had received 33 days of treatment with antituberculosis drugs and 28 days of treatment with oral linezolid for tuberculous meningitis. Severe lactic acidosis was reversed by withdrawing antituberculosis drugs and using continuous venovenous hemodiafiltration (CVVH). When the patient's condition was stable, she was transferred to the infectious disease department, and antituberculosis drugs, with the exception of linezolid, were reintroduced. This did not result in recurrence of lactic acidosis. The causal relationship between lactic acidosis and linezolid was categorized as ‘probable’ on the Adverse Drug Reaction Probability Scale. This case demonstrates that CVVH has potential as an alternative to discontinuation of linezolid alone for rapid reversal of linezolid-associated severe lactic acidosis.     

Expression and activity of thrombomodulin in human gingival epithelium: in vivo and in vitro studies

Epidermal keratinocytes thrombomodulin (TM) has been shown to regulate thrombin at sites of cutaneous injury in addition to a role for epidermal differentiation. TM, a major anticoagulant proteoglycan of the endothelial cell membrane, is a thrombin receptor that acts as a co‐factor for protein C activation. Thrombin has pro‐inflammatory effects for periodontitis. However, little is known about TM in gingival tissue with periodontitis. We used immunohistochemistry to examine expression of TM in gingival epithelium from patients with periodontitis. In vitro , we observed TM expression at varying Ca²⁺ concentrations by confocal laser scanning microscopy, examined the expression of TM mRNA and tested TM co‐factor activity. Furthermore, we measured TM concentration in gingival crevicular fluid (GCF) from 11 severe adult cases of periodontitis using enzyme‐linked immunosorbent assay. Immunoreactive TM was present in gingival epithelium and junctional epithelium, and was reduced in inflamed gingival epithelium compared to healthy gingival epithelium. Ultrastructurally, TM, including microvilli, was observed on the cell membrane. TM localization in cells cultured in 0.09 m m Ca²⁺ differed from that in cells exposed to 1.2 m m Ca²⁺. Northern analysis demonstrated TM mRNA in gingival keratinocytes more than in human umbilical vein endothelial cells (HUVEC). Gingival keratinocytes also facilitated protein C activation by thrombin, although less strongly than HUVEC. TM in GCF at sites with bleeding on probing in patients was significantly elevated ( p <0.001, Student's t ‐test). TM in gingival epithelium may regulate thrombin activity at sites of coagulation and inflammation with periodontal disease, although inflammation may impair this regulation of thrombin.     

Assessment of the Thrombogenic Effect of Fibrin Sealant Dressing in a Vascular Surgery Model in Rabbits

This study's objective was to investigate the potential thrombogenic effects of thrombin-containing fibrin sealant dressings (FSD) in a vascular repair model. Oval-shaped pieces of the rabbit abdominal aorta and vena cava were excised, the injuries were repaired with FSD, and animals were allowed to recover. Thrombus formation was examined by (1) an infusion of indium-labeled platelets into the rabbits following FSD application and estimation of total number of platelets attached to the wounds at 2, 4, and 6 h later (short-term effect, n = 12); and by (2) morphological and histological examinations of the vessels and dressings on days 1, 3, and 7 after repair operation in another group of rabbits (long-term effect, n = 12). Application of FSD sealed the vascular injures and produced immediate hemostasis that was stable up to 1 week. The highest numbers of platelets (both native and labeled) adhered to the arterial and venous repair sites were 6.5 × 10⁶ and 4.4 × 10⁷, respectively, 6 h after operation. The adhered platelets, however, did not form a visible and clinically significant thrombus. In long-term experiments, no evidence of thrombus was found in the lumens of the repaired vessels or on the dressings, and no microthrombi were detected histologically in other tissues at any time point. Although vena caval injuries showed signs of healing at day 7 postoperatively, the aortic wounds expanded progressively (pseudoaneurysm) and were prone to rupture at later times. Thus, direct exposure of FSD does not cause intravascular thrombosis or thrombotic events in rabbits. The dressing appears to be safe and effective for short-term repair of vascular injuries. It may also allow healing of minor venous defects, but cannot replace conventional surgical techniques (suturing) for permanent repair of arterial damages.     

流式细胞术测定保存血小板再表达CD62p方法的建立及应用

目的　①建立并优化测定血小板再表达膜表面激活标志物CD6 2p能力的流式细胞术 (FCM) ;②测定2 2℃保存液体血小板不同保存期再表达CD6 2p的能力 ,以评价该指标在血小板质量监测以及新的保存方法研究中的应用价值。方法　①采用浓度梯度法优化GPRP浓度条件 ,采用析因设计优化凝血酶浓度和 37℃孵育时间条件 ,寻找最佳阴、阳性对照 ;②将 13份血小板按AABB标准保存 ,并延长保存至 12d ,测定每天血小板CD6 2p再表达率 ,与其相应体内存活期作直线相关性分析。结果　①约 1.5× 10 6个血小板内加入 2 .5mmol/LGPRP可有效防止过量凝血酶诱导的聚集反应 ,孵育条件以 1U/L凝血酶 37℃ 15min然后室温置 15min最佳。采用新鲜富含血小板血浆(FPRP)为阴性对照 ,以凝血酶激活FPRP为阳性对照效果良好。② 2 2℃保存血小板CD6 2p再表达率与其相应体内存活期呈明显正相关 ,单份相关系数 (r) 0 .91～ 0 .99,总体相关系数为 0 .99。结论　①优化建立了一种灵敏简便 ,稳定性和重复性并好的流式细胞术方法用于测定保存血小板CD6 2p再表达率 ;②保存液体血小板CD6 2p再表达率与其相应体内存活期呈明显正相关 ,且相关性良好 ,提示CD6 2p再表达率是监测保存血小板质量的良好指标 ,同时还可能在新的保存方法研究和临床血小板功?     

2型糖尿病与凝血功能障碍

目的　探讨2型糖尿病(2DM)患者凝血功能指标的变化,及其与血管并发症的关系。方法　分别检测糖尿病组(n=85 )和对照组(n =6 0 )血浆凝血酶原时间(PT)、活化部分凝血酶原时间(APTT)、凝血酶时间(TT)、纤维蛋白原(FIB)、甘油三酯(TG)、胆固醇(CH)。结果　(1) 2DM组与对照组比较:无论有无血管并发症2DM患者PT、APTT显著下降(P <0 .0 5或P <0 0 1) ,FIB显著上升(P <0 .0 5或P <0 .0 1) ,无血管病变组TT的变化没有统计意义;伴血管病变组患者TT显著下降(P <0 0 1)。(2 ) 2DM组组内比较,伴血管病变组凝血指标PT、APTT、TT均较无血管病变组显著下降(P <0 0 1) ,且FIB水平显著上升(P <0 .0 5 )。(3)DM患者血脂异常的分型比较,DM患者以高甘油三酯血症为主,有血管并发症组的高甘油三酯血症阳性率可达2 8.9% ,但两组差异不明显(P >0 .0 5 ) ;并且TG与四项凝血指标均无明显相关性。结论　2型糖尿病患者存在凝血功能的障碍,且随着血管并发症的出现,患者凝血活性增强。     

Initiating and potentiating role of platelets in tissue factor-induced thrombin generation in the presence of plasma: subject-dependent variation in thrombogram characteristics

Summary.  The hemostatic activity of plasma is determined by platelet activation and coagulation, which processes are mutually stimulatory. We studied this interaction by measuring the cleavage of fluorescent thrombin substrate in platelet-rich plasma (PRP), using the calibrated thrombogram method. In freshly isolated human plasma, thrombin formation triggered by tissue factor was fully dependent on the presence of platelets. It was abolished by annexin A5, indicating dependence on phosphatidylserine (PS) exposure at activated platelets. Comparison of plasmas from various subjects showed considerable interindividual variation in total amount of thrombin generation, regardless of whether platelets or PS-containing phospholipids were present. Integrin α_IIbβ₃ antagonists and ADP receptor blockage, but not aspirin, decreased the rate of thrombin generation (thrombin peak level) and extended the time of onset. Platelet inhibition with cAMP-elevating agents decreased the thrombin-forming rate, but surprisingly shortened the onset time. Stimulation of platelets with agonists of Gi/q-coupled receptors and, to a larger extent, with collagen or Ca²⁺-ionophore increased the rate of thrombin generation and shortened its onset. In PRP from donors with low and high generation, platelet inhibitors and activators were similarly effective. Taken together, these results indicate that, in tissue factor-triggered PRP, PS exposure on activated platelets regulates both onset and rate of thrombin generation. However, coagulant activity rather than platelet activation determines the total amount of thrombin formed, i.e. the endogenous thrombin potential. Thus, kinetics of thrombin generation in PRP are controlled by platelet inhibitors and agonists, but the process is restricted in amount by the subject-dependent variation in coagulation.     

Recombinant factor VIIa enhances platelet adhesion and activation under flow conditions at normal and reduced platelet count.

BACKGROUND: Recombinant factor VIIa (rFVIIa), which was developed for treatment of inhibitor-complicated hemophilia, appears suitable as prohemostatic agent in other clinical disorders including patients with thrombocytopenia. It is generally accepted that rFVIIa functions by enhancement of thrombin generation at the site of injury. It is, however, unknown if and how this affects platelet adhesion and aggregation. OBJECTIVES: To determine the effect of rFVIIa-mediated thrombin generation on platelet adhesion and aggregation under flow conditions at normal and reduced platelet counts. METHODS: Washed platelets and red cells were combined to obtain plasma-free blood with different platelet counts. The reconstituted blood was perfused over a collagen- or fibrinogen-coated surface in the absence or presence of a thrombin generating system consisting of purified coagulation factors rFVIIa, factor (F)X and prothrombin. RESULTS: Addition of coagulation factors rFVIIa, FX and prothrombin to washed platelets and red cells enhanced platelet adhesion and aggregation to collagen and adhesion and spreading to fibrinogen at normal platelet count and at platelet numbers as low as 10 000 microL(-1). rFVIIa-mediated thrombin generation enhanced the activation state of platelets as measured by intracellular calcium fluxes, and enhanced the exposure of procoagulant phospholipids as measured by annexin A5 binding. CONCLUSIONS: Taken together, increased platelet adhesion and aggregation by rFVIIa-mediated thrombin formation may explain the therapeutic effects of rFVIIa in thrombocytopenic conditions and in patients with a normal platelet count by (i) enhancement of primary hemostasis and (ii) enhancement of procoagulant surface leading to elevated fibrin formation.