The initial experience of antithrombin III in the management of neonates with necrotizing enterocolitis

Background

Necrotizing enterocolitis (NEC), the devastating enteric process of premature neonates, is marked by severe intravascular abnormalities and disseminated intravascular coagulation. Treatment to date remains historical and continues to be merely supportive without attempts to ameliorate progress within the inflammatory or coagulation cascades. Antithrombin III (ATIII) supplementation has been shown to favorably alter the process of disseminated intravascular coagulation and sepsis in adults. However, no reported use of this treatment exists in neonates. Therefore, we analyze the efficacy of our recent experience with ATIII replacement therapy in neonates with NEC.

Methods

Age and diseased-matched controls with NEC were identified before the introduction of ATIII in our institution and compared against neonates with NEC undergoing ATIII replacement for diminished ATIII levels. Data collected included demographics, course of treatment parameters, and outcomes. Course of treatment parameters included hemoglobin, platelet count, prothrombin time, and partial thromboplastin time over the first 10 consecutive days of treatment. Outcome variables included packed red blood cell, platelet, fresh frozen plasma, and cryoprecipitate transfusions, as well as transfusion cost, length of stay, and survival.

Results

Over a 5-year period, 19 neonates with NEC received ATIII and were compared to 17 historical controls. Treatment hematologic profiles were not worsened in the ATIII-treated patients. The control patients received less overall transfusions and had a shorter length of stay.

Conclusion

Antithrombin III appears to be safe in neonates with NEC, and its impact on reversing intravascular pathology in these patients warrants more thorough investigation.     

妊娠晚期孕妇抗凝与纤溶活性检测的临床意义

目的:探讨妊娠晚期孕妇血浆中D-二聚体(DD)含量、抗凝血酶(AT)、蛋白C(PC)、蛋白S(PS)活性的变化。方法:分别测定94例妊娠晚期孕妇和27例正常未妊娠妇女血浆中AT、PC、PS的活性以及DD的含量并进行对比分析。结果:妊娠晚期孕妇血浆中DD含量比对照组明显增高,AT、PC、PS活性显著减低。结论:正常妊娠晚期孕妇血浆中抗凝成分AT、PC、PS活性减低、纤溶活性增强,妊娠期检测AT、PC、PS活性、DD含量的变化,对预防出血、降低DIC的发生率具有指导意义,临床应用中各实验室应建立孕妇AT、PC、PS活性及DD含量的参考范围,而不能套用一般健康人的参考范围。     

三种常用指标体系在评估乙型肝炎病毒相关慢加急性肝衰竭患者短期预后中的应用比较

目的探讨预后营养指数（PNI）、抗凝血酶Ⅲ（AT-Ⅲ）、中国重症乙型肝炎研究学组-慢加急性肝衰竭评分（COSSH-ACLFs）模型在乙型肝炎病毒（HBV）相关慢加急性肝衰竭（ACLF）患者病情严重程度及短期预后中的价值。 方法回顾性分析2016年1月至2021年9月在南通大学附属南通第三医院诊治的277例HBV相关ACLF患者的临床资料,根据90 d预后情况,将患者分为好转组（108例）和恶化组（169例）。根据患者入院后24 h内血常规、肝肾功能、凝血功能、肝性脑病分级、平均动脉压、血氧饱和度、吸入氧流量,结合患者年龄,计算出PNI、慢性肝衰竭-慢加急性肝衰竭评分、COSSH-ACLFs。采用Logistic回归分析及受试者工作特征（ROC）曲线评估PNI、AT-Ⅲ、COSSH-ACLFs预测HBV相关ACLF患者90 d短期预后的价值。 结果恶化组COSSH-ACLFs明显高于好转组（Z = 11.189,P<0.001）,PNI、AT-Ⅲ水平均明显低于好转组（Z = 6.815、6.000,P均<0.001）。多因素回归分析结果提示PNI[比值比（OR）= 0.886,95%置信区间（CI）（0.815,0.963）,P = 0.004]、AT-Ⅲ [OR = 0.925,95%CI（0.893,0.958）,P<0.001]、COSSH-ACLFs[OR = 11.456,95%CI（5.700,23.023）,P<0.001]为HBV相关ACLF患者90 d预后的独立影响因素。PNI、AT-Ⅲ、COSSH-ACLFs预测HBV相关ACLF患者短期预后的曲线下面积（AUC）分别为0.737、0.720、0.893;三者联合预测模型的预测效能最佳,AUC达到0.926。 结论PNI、AT-Ⅲ、COSSH-ACLFs对预测HBV相关ACLF患者的短期预后具有良好的价值,联合应用预测价值更优。     

High dose antithrombin III inhibits HMGB1 and improves endotoxin-induced acute lung injury in rats

Objective High mobility group box 1 (HMGB1) is an important factor in the development of sepsis. Previous work suggests that antithrombin III (ATIII) inhibits inflammation, but the mechanism of action is still poorly understood. Design and setting Prospective controlled animal study in a university laboratory. Materials Rats were randomly divided into a lipopolysaccharide (LPS)-induced sepsis control group and an ATIII-treated experimental group. Animals in the experimental group received a bolus of 250 units/kg of ATIII injected into the tail vein. Measurements and results Animals receiving high-dose ATIII (250 units/kg) had significantly improved lung histopathology and survival compared to the control rats. We measured serum and lung levels of various cytokines and HMGB1 at regular intervals from 0 to 12 h after the induction of sepsis and demonstrated lower HMGB1 levels over time in ATIII-treated animals. In an in vitro experiment, we stimulated the mouse macrophage cell line RAW 264.7 with LPS in the presence or absence of ATIII. Subsequent measurement of HMGB1 concentrations in the supernatant and cell signaling molecules in cell lysates revealed an ATIII dose-dependent decrease in HMGB1 release. Furthermore, inhibition of IkB and p42 phosphorylation was observed with the administration of ATIII, suggestive of downstream signaling pathways. Conclusions High-dose ATIII decreases lung pathology and reduces mortality in a rat sepsis model. This finding may be mediated by the inhibition of HMGB1. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. This article is discussed in the editorial available at: .     

Antithrombin III in patients with severe sepsis: a pharmacokinetic study

Objectives: To evaluate the safety, pharmacokinetics, and the practicability of two different antithrombin III (AT III) high-dose regimens in patients with severe sepsis.¶Design: Prospective, open, randomized, 2 parallel groups, multinational clinical trial.¶Setting: Eleven academic medical center intensive care units (ICU) in Austria, Belgium, Denmark, Germany, Norway and Sweden.¶Patients: Thirty-three patients with severe sepsis who received standard supportive care and antimicrobial therapy, in addition to the administration of AT III.¶Interventions: Patients received an intravenous loading dose of 6,000 IU AT III followed by either intermittent bolus infusions of 1,000 IU AT III every 4 h or a continuous infusion of 250 IU AT III/h for 4 days, resulting in a total dose for both dosage regimens of 30,000 IU AT III.¶Measurements: All patients were evaluated for safety and all but one for pharmacokinetics.¶Results and conclusions: The administration of AT III was safe and well tolerated. The overall 28-day all-cause mortality was 30 % (43 % intermittent bolus infusions; 21 % continuous infusion). The mean probability of dying according to the SAPS II was 48 %. The difference in mortality between both groups was within the range of chance. AT III plasma levels were elevated from low baseline levels to above 120 % soon after onset of AT III therapy and remained at these levels for the treatment phase of 4 days. Functional and immunologic levels of AT III corresponded very well. With an overall median volume of distribution of 4.5 l (range: 2.4–6.5 l), AT III only moderately extended beyond plasma. The overall median elimination half-life was 18.6 h (range: 5.1–37.4). Overall, median response was 1.75 % per IU/kg (range: 1.14–2.8).¶The variability of elimination parameters was quite noteworthy (CV = 41–59 %), whereas distribution-related parameters showed a moderate variability (CV = 24 %). In spite of this variability, both high-dose IV regimens reliably provided AT III levels above 120 % for all but one patient. An increased mortality was observed for patients with a distribution volume exceeding 4.5 l (or a response < 1.7 % per IU/kg). AT III distribution volumes above 4.5 l might indicate a capillary leak phenomenon. The continuous infusion regimen was slightly preferred by the investigators with regard to practicability.     

儿童感染性疾病凝血指标检测的临床意义

目的探讨儿童感染性疾病中,凝血功能变化对感染的诊断及转归的意义。方法取患儿感染期标本,分别以凝固法、发色底物法、免疫胶体金法等检测凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、纤维蛋白原含量(FIB)、凝血酶时间(TT)、抗凝血酶活性(AT:A)、D-二聚体(DD)等凝血及抗凝指标。结果感染患儿的凝血指标与正常对照组儿童间差异有统计学意义,特别是AT:A与DD的结果在弥散性血管内凝血(D IC)组与非D IC组中差异有统计学意义。结论各凝血指标特别是AT:A和DD的检测可作为D IC前期诊断和预后评估的指标。     

Impact of an autologous oxygenating matrix culture system on rat islet transplantation outcome

Disruption of the pancreatic islet environment combined with the decrease in oxygen supply that occurs during isolation leads to poor islet survival. The aim of this study was to validate the benefit of using a plasma-based scaffold supplemented with perfluorodecalin to improve islet transplantation outcome.Rat islets were cultured in three conditions: i) control group, ii) plasma based-matrix (P-matrix), and iii) P-matrix supplemented with emulsified perfluorodecalin. After 24 h culture, matrix/cell contacts (Integrinβ1, p-FAK/FAK, p-Akt/Akt), survival (caspase 3, TUNEL, FDA/PI), function, and HIF-1α translocation were assessed. Afterwards, P-matrices were dissolved and the islets were intraportally transplanted. Graft function was monitored for 31 days with glycaemia and C-peptide follow up. Inflammation was assessed by histology (macrophage and granulocyte staining) and thrombin/anti-thrombin complex measurement.Islet survival correlated with an increase in integrin, FAK, and Akt activation in P-matrices and function was maintained. Perfluorodecalin supplementation decreased translocation of HIF-1α in the nucleus and post-transplantation islet structure was better preserved in P-matrices, but a quicker activation of IBMIR resulted in early loss of graft function.“Oxygenating” P-matrices provided a real benefit to islet survival and resistance in vivo. However, intraportal transplantation is not suitable for this kind of culture due to IBMIR; thus, alternative sites must be explored.     

红花黄色素对羟自由基损伤抗凝血酶Ⅲ的保护作用

邻二氮菲-Fe(2+)氧化法及硫代巴比妥酸比色法实验结果表明，红花黄色素清除羟自由基及抑制小鼠肝匀浆脂质过氧化作用的IC50。分别为7.5g／L、5.7×10-2g／L，两作用均呈明显的量效关系。SDS－PAGE电泳结果表明，红花黄色素可抑制羟自由基诱发的兔抗凝血酶Ⅲ解聚。