肺癌患者血浆纤维蛋白原、D-二聚体及抗凝血酶Ⅲ水平的变化及临床意义

目的观察肺癌患者血浆纤维蛋白原(FIB)、D-二聚体及抗凝血酶Ⅲ(AT-Ⅲ)水平变化并探讨其临床意义。方法肺癌患者73例(观察组),健康志愿者60例(对照组),检测两组血浆FIB、D-二聚体及AT-Ⅲ水平。结果观察组血浆FIB、D-二聚体水平显著高于对照组,而AT-Ⅲ水平显著低于对照组(P均<0.05);Ⅲ、Ⅳ期患者血浆FIB、D-二聚体水平高于Ⅰ、Ⅱ期(P<0.05),而Ⅲ、Ⅳ期患者AT-Ⅲ水平低于Ⅰ、Ⅱ期(P<0.05)。结论检测FIB、D-二聚体以及AT-Ⅲ的水平可作为肺癌进展的辅助诊断指标。

Anticoagulation with direct thrombin inhibitors during extracorporeal membrane oxygenation

Use of extracorporeal membrane oxygenation to support patients with critical cardiorespiratory illness is increasing.Systemic anticoagulation is an essential element in the care of extracorporeal membrane oxygenation patients.While unfractionated heparin is the most commonly used agent,unfractionated heparin is associated with several unique complications that can be catastrophic in critically ill patients,including heparin-induced thrombocytopenia and acquired antithrombin deficiency.These complications can result in thrombotic events and subtherapeutic anticoagulation.Direct thrombin inhibitors(DTIs)are emerging as alternative anticoagulants in patients supported by extracorporeal membrane oxygenation.Increasing evidence supports DTIs use as safe and effective in extracorporeal membrane oxygenation patients with and without heparininduced thrombocytopenia.This review outlines the pharmacology,dosing strategies and available protocols,monitoring parameters,and special use considerations for all available DTIs in extracorporeal membrane oxygenation patients.The advantages and disadvantages of DTIs in extracorporeal membrane oxygenation relative to unfractionated heparin will be described.

高凝状态若干临床方面的进展

本文综述高凝状态的病因、发病率、临床表现及其实验室筛选检查,评价其临床意义和实用价值,并详述防治措施。

急性脑梗死患者OCSP分型与凝血指标变化相关性研究的临床意义

目的：探讨急性脑梗死患者OCSP分型与凝血酶原片段1＋2和抗凝血酶3的相关性。方法：对入住我院神经内二科的64例患急性脑梗死患者进行OCSP分型，并选择门诊健康体检者作对照。应用酶联免疫吸附法（ ELISA）进行检测所有患者的血浆凝血酶原片段1＋2浓度和抗凝血酶3活性；分析两者与急性脑梗死OCSP分型的相关性。结果：急性脑梗死各亚型中，完全前循环性脑梗死（ TACI）、部分前循环性脑梗死（ PACI）和后循环性脑梗死（ POCI）患者血浆凝血酶原片段1＋2浓度明显高于腔隙性脑梗死（ LACI）和健康对照者，抗凝血酶3活性明显低于腔隙性脑梗死和健康对照者。结论：血浆凝血酶原片段1＋2浓度和抗凝血酶3活性的增高与TACI及PACI密切相关。

Antithrombin reduces reperfusion-induced hepatic metastasis of colon cancer cells

AIM： To examine whether antithrombin （AT） could prevent hepatic ischemia/reperfusion （I/R）-induced hepatic metastasis by inhibiting tumor necrosis factor （TNF）-α-induced expression of E-selectin in rats. METHODS： Hepatic I/R was induced in rats and mice by clamping the left branches of the portal vein and the hepatic artery. Cancer cells were injected intrasplenically. The number of metastatic nodules was counted on day 7 after I/R. TNF-α and E-selectin mRNA in hepatic tissue, serum fibrinogen degradation products and hepatic tissue levels of 6-keto-PGF1α, a stable metabolite of PGI2, were measured. RESULTS： AT inhibited increases in hepatic metastasis of tumor cells and hepatic tissue mRNA levels of TNF-α and E-selectin in animals subjected to hepatic I/R. Argatroban, a thrombin inhibitor, did not suppress any of these changes. Both AT and argatroban inhibited I/R-induced coagulation abnormalities. I/R-induced increases of hepatic tissue levels of 6-keto-PGF1α were significantly enhanced by AT. Pretreatment with indomethacin completely reversed the effects of AT. Administration of OP-2507, a stable PGI2 analog, showed effects similar to those of AT in this model. Hepatic metastasis in congenital AT-deficient mice subjected to hepatic I/R was significantly increased compared to that observed in wild-type mice. Administration of AT significantly reduced the number of hepatic metastases in congenital AT-deficient mice. CONCLUSION： AT might reduce I/R-induced hepatic metastasis of colon cancer cells by inhibiting TNF-α-induced expression of E-selectin through an increase in the endothelial production of PGI2. These findings also raise the possibility that AT might prevent hepatic metastasis of tumor cells if administered during the resection of liver tumors.

Anticoagulant modulation of inflammation in severe sepsis

Inflammation and coagulation are so tightly linked that the cytokine storm which accompanies the development of sepsis initiates thrombin activation and the development of an intravascular coagulopathy. This review examines the interaction between the inflammatory and coagulation cascades, as well as the role of endogenous anticoagulants in regulating this interaction and dampening the activity of both pathways. Clinical trials attempting to improve outcomes in patients with severe sepsis by inhibiting thrombin generation with heparin and or endogenous anticoagulants are reviewed. In general, these trials have failed to demonstrate that anticoagulant therapy is associated with improvement in mortality or morbidity. While it is possible that selective patients who are severelyill with a high expected mortality may be shown to benefit from such therapy, at the present time none of these anticoagulants are neither approved nor can they be recommended for the treatment of sepsis.

Antithrombin in the treatment of burn trauma

Antithrombin(AT) is a natural anticoagulant with antiinflammatory properties that has demonstrated value in sepsis, disseminated intravascular coagulation and in burn and inhalation injury. With high doses, AT maydecrease blood loss during eschar excision, reducing blood transfusion requirements. There are no human randomized, placebo-controlled studies, which have tested the true benefit of this agent in these conditions. Two main forms of AT are either plasma-derived AT(ph AT) and recombinant AT(rh AT). Major ovine studies in burn and smoke inhalation injury have utilized rh AT. There have been no studies which have either translated the basic rh AT research in burn trauma, or determined the tolerance and pharmacokinetics of rh AT concentrate infusions in burn patients. Advantages of rh AT infusions are no risk of blood borne diseases and lower cost. However, the majority of human burn patient studies have been conducted utilizing ph AT. Recent Japanese clinical trials have started using ph AT in abdominal sepsis successfully. This review examines the properties of both ph AT and rh AT, and analyzes studies in which they have been utilized. We believe that it is time to embark on a randomized placebo-controlled multi-center trial to establish the role of AT in both civilian and military patients with burn trauma.

Antithrombin Ⅲ injection via the portal vein suppresses liver damage

AIM:To investigate the effects of antithrombin Ⅲ(AT Ⅲ) injection via the portal vein in acute liver failure.METHODS:Thirty rats were intraperitoneally challenged with lipopolysaccharide(LPS) and D-galactosamine(GalN) and divided into three groups:a control group;a group injected with AT Ⅲ via the tail vein;and a group injected with AT Ⅲ via the portal vein.AT Ⅲ(50 U/kg body weight) was administrated 1 h after challenge with LPS and GalN.Serum levels of inflammatory cytokines and fibrin degradation products,hepatic fibrin deposition,and hepatic mRNA expression of hypoxiarelated genes were analyzed.RESULTS:Serum levels of alanine aminotransferase,tumor necrosis factor-α and interleukin-6 decreased significantly following portal vein AT Ⅲ injection compared with tail vein injection,and control rats.Portal vein AT Ⅲ injection reduced liver cell destruction and decreased hepatic fibrin deposition.This treatment also significantly reduced hepatic mRNA expression of lactate dehydrogenase and heme oxygenase-1.CONCLUSION:A clinically acceptable dose of AT Ⅲ injection into the portal vein suppressed liver damage,probably through its enhanced anticoagulant and antiinflammatory activities.

Predicting early outcomes of liver transplantation in young children: The EARLY study

AIM To determine potentially modifiable predictors of early outcomes after liver transplantation in children of age < 3 years.METHODS This study was a retrospective chart review including all consecutive children of age less than 3-years-old having had a liver transplant done at the Western Canadian referral center from June 2005 to June 2015.Pre-specified potential predictor variables and primary and secondary outcomes were recorded using standard definitions and a case report form. Associations between potential predictor variables and outcomes were determined using univariate and multiple logistic [odds ratio(OR); 95%CI] or linear(effect size, ES; 95%CI) regressions. RESULTS There were 65 children, of mean age 11.9(SD 7.1) mo and weight 8.5(2.1) kg, with biliary-atresia in 40(62%), who had a living related donor [LRD; 29(45%)], split/reduced [21(32%)] or whole liver graft [15(23%)]. Outcomes after liver transplant included: ventilator-days of 12.5(14.1); pediatric intensive care unit mortality of 5(8%); re-operation in 33(51%), hepatic artery thrombosis(HAT) in 12(19%), portal vein thrombosis(PVT) in 11(17%), and any severe complication(HAT, PVT, bile leak, bowel perforation, intraabdominal infection, retransplant, or death) in 32(49%) patients. Predictors of the prespecified primary outcomes on multiple regression were:(1) HAT: split/reduced(OR 0.06; 0.01, 0.76; P = 0.030) or LRD(OR 0.16; 0.03, 0.95; P = 0.044) vs whole liver graft; and(2) ventilator-days: surgeon(P < 0.05), lowest antithrombin(AT) postoperative day 2-5(ES-0.24;-0.47,-0.02; P = 0.034), and split/reduced(ES-12.5;-21.8,-3.2; P = 0.009) vs whole-liver graft. Predictors of the pre-specified secondary outcomes on multiple regression were:(1) any thrombosis: LRD(OR 0.10; 0.01, 0.71; P = 0.021) or split/reduced(OR 0.10; 0.01, 0.85; P = 0.034) vs whole liver graft, and lowest AT postoperative day 2-5(OR 0.93; 0.87, 0.99; P = 0.038); and(2) any severe complication: surgeon(P < 0.05), lowest AT postoperative day 2-5(OR 0.92; 0.86-0.98; P = 0.016), and split/reduced(OR 0.06; 0.01, 0.78; P = 0.032) vs whole-liver graft. CONCLUSION In young children, whole liver graft and surgeon was associated with more complications, and higher AT postoperative day 2-5 was associated with fewer complications early after liver transplantation.

Impact of antithrombin Ⅲ on hepatic and intestinal microcirculation in experimental liver cirrhosis and bowel inflammation: An in vivo analysis

AIM： To analyze the hepatic and intestinal microcirculation in an animal model of liver cirrhosis and inflammatory bowel disease （IBD） and to characterize bhe anti-inflammatory action of antithrombin Ⅲ （ATⅢ） on leukocyte kinetics and liver damage. METHODS： Hepatic and intestinal microcirculation was investigated by intravital videomicroscopy. Standardized models of experimental chronic liver cirrhosis and bowel inflammation were employed. Animals were divided into four groups （n = 6/group）： controls, animals with cirrhosis, animals with cirrhosis and IBD, animals with cirrhosis and IBD treated with ATIII. RESULTS： Cirrhosis facilitated leukocyte rolling and sticking in hepatic sinusoids （1.91±0.28 sticker/μm vs 0.5±0.5 sticker/μm in controls, P〈0.05）. The effect enhanced in animals with cirrhosis and IBD （5.4±1.65 sticker/μm）, but reversed after ATIII application （3.97±1.04 sticker/μm, P〈0.05）. Mucosal blood flow showed no differences in cirrhotic animals and controls （5.3±0.31 nL/min vs5.4±0.25 nL/min） and was attenuated in animals wibh cirrhosis and IBD significantly （3.49±0.6 nL/min）. This effect was normalized in the treatment group （5.13±0.4 nL/min, P〈0.05）. Enzyme values rose during development of cirrhosis and bowel inflammation, and reduced after ATIII application （P〈0.05）. CONCLUSION： Liver cirrhosis in the presence of IBD leads to a significant reduction in mucosal blood flow and an increase in hepatic leukocyte adherence with consecutive liver injury, which can be prevented by administration of ATⅢ.

Antithrombin gene Arg197Stop mutation-associated venous sinus thrombosis in a Chinese family

This study sought to elucidate the genetic correlation of cerebral venous sinus thrombosis caused by a hereditary antithrombin deficiency in a Chinese family, at the genetic and protein levels. A nonsense mutation from C to T on locus 6431 in exon 3B of the antithrombin gene was observed, leading to an arginine （CGA） to stop codon （TGA） change in the protein. This is the first report of this mutation in China. Ineffective heparin therapy in the propositus patient is associated with a lack of heparin binding sites after antithrombin gene mutation. Characteristic low intracranial pressure in the acute phase might be specific to this patient with cerebral venous sinus thrombosis.

Heparin and other anticoagulants in amniotic fluid embolism(AFE):Literature review and concept of the therapy

Aim: The objective of this study is to review all the reported outcomes of heparin application in amniotic fluid embolism (AFE) so far and to find out why, when and how heparin or other anticoagulants should be used in AFE. Material and methods: We searched Medline (from 1969 to 2011), using two key words: 1) amniotic fluid embolism;2) amniotic fluid embolism and heparin. The search for the former produced 1127 replies, of which 208 were case reports of AFE. In response to the other key word, there were 94 articles. We looked through all the articles, selecting those relevant for our study. Results: In the years 1969-2011, 208 AFE cases were reported. Heparin (unfractionated heparin) or low molecular weight heparin (LMWH) was applied in 20 cases (9.6%), being the main drug in 11 cases (5.3%) and in 6 cases as a component of spectacular treatment of AFE (surgical treatment and extracorporal membrane oxygenation). In one of these cases anithrombin (AT) with LMWH was used. In one patient heparin therapy was considered to be unsuccessful and hence recombinant plasminogen activator (rt-PA) was instituted. All the patients survived. Conclusions: 1) The attempts to use heparin in AFE could be defined as promising, although the number of treated patients is too small for conclusion;2) The postulate to use heparin at the very onset of AFE (a bolus of 10,000 U followed by monitored intravenous infusion) has serious justification: one of the pathways of AFE is the target for heparin (coagulation pathway).

Effects of fused hirudin on activity of thrombin and function of platelets

Objective: To investigate whether fused hirudin peptide has both antithrombin and antiplatelet functions. Methods: The core region of fused hirudin was the C-terminal tail of hirudin(hirudin_ 53-64),which could bind to the anion binding exosite (ABE) of thrombin.Arg-Pro-Pro-Gly-Phe(RPPGF) amino acid sequence,a metabolite of bradykinin,was added to the N-terminus of hirudin_ 53-64.It bound to the active site of thrombin.Additionally,Arg-Gly-Asp(RGD)amino acid sequence,an inibitor of glycoprotein Ⅱb/Ⅲa( GP Ⅱb/Ⅲa) receptor,was linked to C-terminus of hirudin_ 53-64.This 26-animo acid-fused hirudin peptide was artificially synthesized,purified and analysed. Results: Fused hirudin peptide significantly lengthened the activated partial thromboplastin time(APTT),thrombin time(TT)and prothrombin time(PT) and inhibited the amidolytic activity of thrombin.The ADP-induced platelet aggregation was markedly inhibited by fused hirudin peptide. Conclusion: Fused hirudin peptide has activity of antithrombin as well as antiplatelet.Therefore bifunctional anticoagulation peptide has capacity to target various components of haemostatic process and may become more powerful antithrombosis agent.

High Performance Affinity Chromatography of Antithrombin III Based on Monodisperse Poly (glycidyl methacrylate) Beads

A new approach for the separation of antithrombin III with high performance affinity chromatography (HPAC) was described. A novel monodisperse, non-porous, cross-linked poly (glycidyl methacrylate) beads (PGMA) were used as the affinity support. With the water-soluble carbodiimide, heparin was linked covalently to amino-PGMA-beads, which was prepared by amination of PGMA. The adsorbent obtained exhibits high binding activity to antithrombin III (ATIII), good resolution and excellent mechanical properties and can be used under high flow rate.

Pharmacokinetic and Pharmacodynamic Properties of Batifiban Coadministered with Antithrombin Agents in Chinese Healthy Volunteers

The combined use of batifiban, a synthetic platelet GP II b/IIIa receptor antagonist, and an- tithrombin agents is an attractive option for the treatment of patients with non-ST-segment elevation （NSTE） acute coronary syndrome （ACS） and those scheduled for percutaneous coronary intervention. To observe whether antithrombin agents affect the pharmacokinetic and pharmacodynamic properties of batifiban in combination therapy and optimize clinical administration dosage of batifiban, an open-label and parallel study was conducted. Thirty healthy subjects were randomly divided into three groups, which were sequentially treated with batifiban alone, or oral coadministration of clopidogrel, aspirin and UFH, or batifiban coadministered with these antithrombin agents. Blood samples were collected at pre-specified time points. The evaluation index included the inhibition of platelet aggregation and pharmacokinetic parameters. The pharmacokinetic parameters of batifiban and batifiban coadministered with antithrombin agents showed no significant differences. The mean inhibition rate of platelet aggre- gation （%） suggested that neither batifiban alone nor antithrombin agents alone could provide such po- tent inhibition rate （〉80%） to obtain the best clinical efficacy, but they had a synergistic effect on plate- let inhibition. No serious adverse effects were observed. The results in these healthy subjects suggest that batifiban coadministrated with antithrombin agents could achieve optimum clinical treatment effect for patients with NSTE ACS, and also those scheduled for percutaneous coronary intervention.

Change of Coagulation Factor Ⅷ and Antithrombin Ⅲ Activity in Bank-Stored Blood

Coagulation factor Ⅷ and antithrombin Ⅲ activity were detected in 15 health donors. It was found that antithrombin Ⅲ activity decreased obviously 12 h after blood drawing. It lost 56 % of the activity at the 3rd day, and 70 % of the activity at the 7th day. FⅧ:c showed no obvious change after 24 h, until the 3rd day. It lost 40 %-60 % of the activity after 36 h and was reduced to the 30 % of the original activity at the 5th day. Our results suggested that at the 3rd day coagulation factor Ⅷ of bank stored blood can be used to replenish antithrombin Ⅲ, while bank stored blood in one day can be used to replenish FⅧ.

DETERMINATION OF PROTEIN C, PROTEIN S ANTIGENS AND THEIR CLINICAL SIGNIFICANCE

PC and PS antigens were measured in 370 cases of 12 different diseases and 14 women at third trimester of pregnancy. The PC level was found significantly raised in coronary heart disease, diabetes, nephrotic syndrome, stroke, aplastic anemia and the third trimester of pregnancy, decreased in chronic hepatitis, acute fulminant hepatitis, cirrhosis and deep vein thrombosis (DVT) and no change in acute hepatitis, primary liver carcinoma, Buerger’s disease and leukemia. Total PS was increased in type Ⅱdiabetes, acute lymphocytic leukemia and DVT, and decreased in Berger’s disease, liver diseases, aplastic anemia and the third trimester of pregnancy. In majority of the cases, the total PS and free PS changed parallelly except in Buerger’s disease, DVT and preeclampsia. No correlation was found between PC and PS, PC and AT-Ⅲ.

Antithrombin Ⅲ deficiency in a patient with recurrent venous thromboembolism:A case report

BACKGROUND Antithrombin Ⅲ(AT3)deficiency,an autosomal dominant disease,increases the likelihood of an individual developing venous thromboembolism(VTE).Longterm anticoagulation treatment is required for those suffering from AT3 deficiency.CASE SUMMARY A man aged 23,who had a history of deep venous thrombosis(DVT),experienced recurrent pain and swelling in his right lower extremity for three days following withdrawal of Rivaroxaban.He was diagnosed with DVT and antithrombin Ⅲ deficiency as genetic testing revealed a single nucleotide variant in SERPINC1(c.667T>C,p.S223P).The patient was advised to accept long-term anticoagulant therapy.CONCLUSION Inherited AT3 deficiency due to SERPINC1 mutations results in recurrent VTE.Patients may benefit from long-term anticoagulant therapy.

Studies on Extraction and Activity of Peptides from Haemadipsa hainana

[Objectives]Peptides was extracted from Haemadipsa hainana and its activity was studied.[Methods]Electric stimulation,water extraction and ultrasonic extraction were used to extract the peptides from H.hainana.Then the protein content and molecular weight distribution of H.hainana peptides were detected by the BCA method and SDS-PAGE method,respectively.The antithrombin activity and analgesic activity of the three peptide extracts of H.hainana were detected by Markwardt thrombin titration method and mouse hot plate experiment,respectively.[Results]There extraction methods of electric stimulation,water extraction and ultrasonic extraction were used to extract the peptide extract of H.hainana,and the yields were as follows:water extraction>electrical stimulation>ultrasonic extraction.The three peptide extracts from H.hainana had antithrombin activity,and the antithrombin activity was as follows:water extraction>ultrasonic extraction>electrical stimulation.Through the hot plate experiment in mice,it was verified that the three peptide extracts of H.hainana had analgesic activity,and the analgesic activity was water extraction>electric stimulation>ultrasonic extraction.The analgesic activity of high-dose(100 mg/kg)group of H.hainana obtained by water extraction was slightly weaker than that of tramadol.[Conclusions]This study confirmed that the peptide extract of H.hainana had certain antithrombin and analgesic activity,laying a foundation for the subsequent development and application of H.hainana.