Relationship between Acquired Deficiency of Vitamin K-dependent Clotting Factors And Hemorrhage

This study examined the changes of activities of vitamin K-dependent clotting factors(VKDCF) under various pathological conditions and explored the relationship between acquired deficiency of VKDCFs and hemorrhage.Clinical data of 35 patients who were diagnosed as having acquired deficiency of VKDCF were retrospectively analyzed.Coagulation factors involved in the intrinsic and extrinsic pathways were detected in these patients and 41 control subjects.The results showed that the average activities of VKDCFs were decreased in the patients in comparison to the control subjects and significantly increased after treatment of these patients with vitamin K and blood products.Multivariate regression analysis indicated that decreased activity of VKDCF was not an independent risk factor for bleeding disorders owing to deficiency or metabolic disturbance of vitamin K.It was concluded that acquired deficiency of VKDCF occurs under a variety of pathologic conditions and is closely associated with hemorrhagic events.Administration of vitamin K and transfusion of blood products containing high concentrations of VKDCFs helps alleviate the hemorrhagic diseases.

A comprehensive method to explore inhibitory kinetics and mechanisms of an anticoagulant peptide derived from Crassostrea gigas

A comprehensive method was applied to evaluate the anticoagulant activity of a novel anticoagulant peptide(NAESLRK)derived from oyster(Crassostrea gigas).The anticoagulant peptide drastically reduced the extrinsic clotting activity and also impaired the intrinsic clotting activity slightly.Consistent with clotting data,the thrombin peak height was reduced to 84.7 nmol/L from 123.4 nmol/L,and thrombin generation time was delayed to 4.67 min from 4.42 min when the extrinsic trigger was applied.The inhibitory kinetics of FⅪa,FⅨa,FⅩa,FⅡa,and APC in a purified component system rationally explained the reduction of extrinsic clotting activity and impairment of thrombin generation.Besides the inhibition of FⅩa and FⅡa activity,the activation processes of FⅩand FⅡby intrinsic/extrinsic tenase complex and prothrombinase were also damaged.The anticoagulant activity in the plasma system was the result of comprehensive inhibition of various factors.The research provided a novel method for anticoagulant evaluation and inhibitory mechanism of bioactive peptides from food products.

Factors predisposing to thrombosis after major joint arthroplasty

BACKGROUND Total joint arthroplasty is one of the most common options for end stage osteoarthritis of major joints.However,we must take into account that thrombosis after hip/knee arthroplasty may be related to mutations in genes encoding for blood coagulation factors and immune reactions to anticoagulants[heparininduced thrombocytopenia(HIT)/thrombosis].Identifying and characterizing genetic risk should help to develop diagnostic strategies or modify anticoagulant options in the search for etiological mechanisms that cause thrombophilia following major orthopedic surgery.AIM To evaluate the impact of patients’coagulation profiles and to study specific pharmacologic factors in the development of post-arthroplasty thrombosis.METHODS In 212(51 male and 161 female)patients that underwent primary total hip arthroplasty(100)or total knee arthroplasty(112)due to osteoarthritis during a period of 1 year,platelet counts and anti-platelet factor 4(PF4)/heparin antibodies were evaluated pre/postoperatively,and antithrombin III,methylenetetrahydrofolate reductase,factor V and prothrombin gene mutations were evaluated preoperatively.In a minimum follow-up of 3 years,196 patients receiving either low-molecular-weight heparins(173)or fondaparinux(23)were monitored for the development of thrombocytopenia,anti-PF4/heparin antibodies,HIT,and thrombosis.RESULTS Of 196 patients,32 developed thrombocytopenia(nonsignificant correlation between anticoagulant type and thrombocytopenia,P=0134.)and 18 developed anti-PF4/heparin antibodies(12/173 for low-molecular-weight heparins and 6/23 for fondaparinux;significant correlation between anticoagulant type and appearance of antibodies,P=0.005).Odds of antibody emergence:8.2%greater in patients receiving fondaparinux than low-molecular-weight heparins.Gene mutations in factor II or V(two heterozygotes for both factor V and II)were identified in 15 of 196 patients.Abnormal low protein C and/or S levels were found in 3 of 196(1.5%)patients,while all patients had normal levels of von Willebrand factor,lupus anticoagulant,and antithrombin III.Four patients developed HIT(insignificant correlation between thrombocytopenia and antibodies)and five developed thrombosis(two had positive antibodies and two were heterozygotes for both factor II&V mutations).Thrombosis was not significantly correlated to platelet counts or HIT.The correlation of thrombosis to antibodies,factor II,factor V was P=0.076,P=0.043,P=0.013,respectively.CONCLUSION Screening of coagulation profile,instead of platelet monitoring,is probably the safest way to minimize the risk of post-arthroplasty thrombosis.In addition,fondaparinux can lead to the formation of anti-PF4/heparin antibodies or HIT.

The Relationship between Hyperhomocysteinemia,Haemostatic Factors and Acute Coronary Syndrome in Southeastern Turkey:A Prospective,Comparative Study

Aim: We investigated the relationship between hyperhomocysteinemia, coagulation factors and acute coronary syndrome. Materials and method: The study was conducted at cardiology and hematology department of Dicle University Medical School between January 1st 2003 and May 31st 2009. The study included 96 patients with acute coronary syn-drome and 96 controls. Results: Baseline characteristics of patients (63 males, 33 females, mean age 56.4 years) and controls (58 males and 38 females, mean age: 51.1 years) were similar. There was a statistically significant difference between two groups according to homocysteine levels (13.4 ±8.0 micromole/L vs. 12.8 ± 7.1 micromole/L p = 0.042). In this study, we found that hyperhomocysteinemia, smoking, elevated levels of CRP, low levels of HDL, positive family history, presence of hypertension, BMI > 27, low levels of protein C and protein S were associated with high risk for acute coronary syndrome. Fibrinogen level, factor V level, factor VIII level, factor IX level ,factor X level ,and factor V leiden (p = 0.128) are not risk factors for acute coronary syndrome. Conclusion: Hyperhomocysteine is a significant risk factor for acute coronary syndrome There is not relationship between coagulation factors and acute coronary syn-drome except low levels of protein C and protein S.

Short-term outcome of total knee replacement in a patient with hemophilia:A case report and review of literature

BACKGROUND Hemophilia A is a rare inherited bleeding disorder caused by mutations in the factor Ⅷ gene. This clotting factor plays an intrinsic role in the blood coagulation pathway. Patients with hemophilia may develop orthopedic manifestations such as hemarthrosis, but multiple malunion of fractures over the knee is rare and difficult to treat.CASE SUMMARY We report a patient with hemophilia A who developed severe knee osteoarthritis along with fracture malunion and nonunion. Total knee replacement was performed using a custom-made modular hinged knee prosthesis(cemented) equipped with extended distal and proximal stems. At 3 years’ follow-up, the patient exhibited excellent clinical function and remained satisfied with the surgical outcome. Surgical intervention was accompanied by rigorous coagulation factor replacement.CONCLUSION This case highlights various unique scenarios specific to individuals with hemophilia and fracture deformity.

Expression and Purification of Human Coagulation Factor X in Mammalian CHO-DG44 Cells

[Objectives]This study was conducted to obtain a Chinese hamster ovary cell line that stably expresses recombinant human coagulation factor X(rhFX),and to induce efficient expression of the target gene with different concentrations of methotrexate(MTX).[Methods]PCR was performed to obtain the rhFX gene,and a recombinant expression plasmid pOptiVEC-rhFX was constructed and subjected to double restriction endonuclease digestion and sequencing identification.CHO-DG44(DHFR-)cells were transfected by the liposome method,and the target protein was purified by affinity chromatography and detected by SDS-PAGE electrophoresis and Western blot.A cell line with efficient and stable expression of the target gene was obtained by increasing the concentration of MTX to select positive clones.[Results]PCR yielded a 1509 bp rhFX sequence,and the results of double digestion and sequencing showed that the constructed pOptiVEC-rhFX plasmid was correct.After transfection of cells,MTX significantly increased protein expression.When MTX reached 1.0μmol/L,the expression efficiency of the target protein was(9±0.27)μg/ml.The purity of the target protein purified by affinity chromatography was 93%,which could be used for subsequent experiments.The expression efficiency of rhFX in eukaryotic mammalian cells was improved by increasing MTX concentration,and an affinity chromatography purification process for the target protein was preliminarily established.[Conclusions]The results of this study provide data support for the expression and purification of rhFX,and will lay a solid foundation for the development of drugs related to rhFX.

Removal selectivity of Prometheus:A new extracorporeal liver support device

AIM： To evaluate whether treatment with the Prometheus system significantly affects cytokines, coagulation factors and other plasma proteins. METHODS： We studied nine patients with acute-onchronic liver failure and accompanying renal failure. Prometheus therapy was performed on 2 consecutive days for up to 6 h in all patients. Several biochemical parameters and blood counts were assessed at regular time points during Prometheus treatment. RESULTS： We observed a significant decrease of both protein-bound （e.g. bile acids） and water-soluble （e.g. ammonia） substances after Prometheus therapy. Even though leukocytes increased during treatment （P〈 0.01）, we found no significant changes of C-reactive protein, interleukin-6, and tumor necrosis factor-o plasma levels （all P 〉 0.5）. Further, antithrombin 3, factor II and factor V plasma levels did not decrease during Prometheus therapy （all P 〉0.5）, and the INR remained unchanged （P = 0.4）. Plasma levels of total protein, albumin, and fibrinogen were also not altered during Prometheus treatment （all P 〉 0.5）. Finally, platelet count did not change significantly during therapy （P= 0.6）. CONCLUSION： Despite significant removal of protein- bound and water-soluble substances, Prometheus therapy did not affect the level of cytokines, coagulation factors or other plasma proteins. Thus, the filters and adsorbers used in the system are highly effective and specific for water-soluble substances and toxins bound to the albumin fraction.

Effects on coagulation factor production following primary hepatomitogen-induced direct hyperplasia

AIM： To investigate the molecular mechanisms involved in coagulation factor expression and/or function during direct hyperplasia （DH）-mediated liver regeneration. METHODS： Direct hyperplasia-mediated liver regeneration was induced in female C57BL/6 mice by administering 1,4-bisr2-（3,5-dichloropyridyloxy）] benzene （TCPOBOP）, a representative hepatomitogen. Mice were weighed and sacrificed at various time points [Day 0 （D0： prior to injection）, 3 h, D1, D2, D3, and D10] after TCPOBOP administration to obtain liver and blood samples. Using the RNA samples extracted from the liver, a comprehensive analysis was performed on the hepatic gene expression profiling of coagulation-related factors by real-time RT-PCR （fibrinogen, prothrombin, factors Ⅴ, Ⅶ, Ⅷ, Ⅸ, Ⅹ, Ⅺ, Ⅻ, ⅩⅢβ , plasminogen, antithrombin, protein C, protein S, ADAMTS13, and VWF）. The corresponding plasma levels of coagulation factors （fibrinogen, prothrombin, factors Ⅴ, Ⅶ, Ⅷ, Ⅸ, Ⅹ, Ⅺ, Ⅻ, ⅩⅢ, and VWF） were also analyzed and compared with their mRNA levels. RESULTS： Gavage administration of TCPOBOP （3 mg/kg body weight） resulted in a marked and gradual increase in the weight of the mouse livers relative to the total body weight to 220% by D10 relative to the DO （control） ratios. At the peak of liver regeneration （D1 and D2）, the gene expression levels for most of the coagulationrelated factors （fibrinogen, prothrombin, factors Ⅴ, Ⅶ, Ⅷ, Ⅸ,Ⅺ, Ⅻ, ⅩⅢβ, plasminogen, antithrombin, protein C, ADAMTS13, VWF） were found to be downregulated in a time-dependent manner, and gradually recovered by D10 to the basal levels. Only mRNA levels of factor X and protein S failed to show any decrease during the regenerative phase. As for the plasma levels, 5 clotting factors （prothrombin, factors Ⅷ, Ⅸ, Ⅺ, and Ⅻ） demonstrated a significant decrease （P 〈 0.05） during the regeneration phase compared with DO. Among these 5 factors, factor Ⅸ and factor Ⅺ showed the most dramatic decline in their activities by about 50% at D2 compared to the basal levels, and these reductions in plasma activity for both factors were consistent with our RT-PCR findings. In contrast, the plasma activities of the other coagulation factors （fibrinogen, factors Ⅴ, Ⅶ, ⅩⅢ, and VWF） were not significantly reduced, despite the reduction in the liver mRNA levels. Unlike the other factors, FX showed a temporal increase in its plasma activity, with significant increases （P 〈 0.05） detected at DI. CONCLUSION： Investigating the coagulation cascade protein profiles during liver regeneration by DH may help to better understand the basic biology of the liver under normal and pathological conditions.

Binding of EGF1 Domain Peptide in Coagulation Factor Ⅶ with Tissue Factor and Its Implications for the Triggering of Coagulation

The binding function of EGF1 domain peptide with tissue factor(TF)and its ability of triggering coagulation were explored.The TF expression model in vitro was established by lipopolysaccha-ride induction.The affinity of EGFP-EGF1 and TF expressing cells was analyzed by fluorescence microscopy and flow cytometry(FCM).The affinity of EGFP-EGF1 and rat soluble TF was quantitated by surface plasmon resonance(SPR).The ability of EGFP-EGF1 in triggering coagulation was tested by prothrombin time assay.The FCM res...

Polymorphisms in the genes for coagulation factor II,V,VII in patients undergoing coronary angiography

Objective: To determine whether polymorphisms in the genes for coagulation factor II,V, VII could predispose an individual to increase risk for coronary artery disease (CAD) and/or myocardial infarction (MI) in Chinese. Methods: We screened coagulation factor II(G20210A),V(G1691A),VII (R353Q and HVR4) genotype in 374 patients undergoing coronary angiography by polymerase chain reaction and restriction fragment length polymorphism (PCR RFLP) assay. Results: The R353Q and HVR4 genotype of the factor VII distribution was in accordance with Hardy Weinberg equilibrium. The frequencies of FVII genotype or allele did not show statistically significant differences between CAD group and controls or between male and female. The frequencies of the Q allele and (RQ+QQ) genotype were significantly higher among the CAD patients without myocardial infarction (MI) history than among those with MI history ( P <0.05). However, HVR4 polymorphism was not significantly different within groups. We only find one normal control of factorII(G20210A) mutation. No coagulation factor V(G1691A) mutation was found in the CAD patients and controls. Conclusion: The factor II(G20210A),V(G1691A) mutation is absent and may not be a major genetic factor for CAD and/or MI; the Q allele of the R353Q polymorphism of the factor VII gene may be a protective genetic factor against myocardial infarction in Chinese.

Acquired coagulation dysfunction resulting from vitamin Kdependent coagulation factor deficiency associated with rheumatoid arthritis: A case report

BACKGROUND Rheumatoid arthritis(RA)is a common chronic inflammatory autoimmune disease with the main clinical feature of progressive joint synovial inflammation,which can lead to joint deformities as well as disability.RA often causes damage to multiple organs and systems within the body,including the blood hemostasis system.Few reports have focused on acquired coagulation dysfunction resulting from vitamin K-dependent coagulation factor deficiency associated with RA.CASE SUMMARY A 64-year-old woman with a history of RA presented to our hospital,complaining of painless gross hematuria for 2 wk.Blood coagulation function tests showed increased prothrombin time,international normalized ratio,and activated partial thromboplastin time.Abnormal blood coagulation factor(F)activity was detected(FII,7.0%;FV,122.0%;and FX,6.0%),indicating vitamin K-dependent coagulation factor deficiency.Thromboelastography and an activated partial thromboplastin time mixed correction experiment also suggested decreased coagulation factor activity.Clinically,the patient was initially diagnosed with hematuria,RA,and vitamin K-dependent coagulation factor deficiency.The patient received daily intravenous administration of vitamin K120 mg,etamsylate 3 g,and vitamin C 3000 mg for 10 d.Concurrently,oral leflunomide tablets and prednisone were administered for treatment of RA.After the treatment,the patient's symptoms improved markedly and she was discharged on day 12.There were no hemorrhagic events during 18 mo of follow-up.CONCLUSION RA can result in vitamin K-dependent coagulation factor deficiency,which leads to acquired coagulation dysfunction.Vitamin K1 supplementation has an obvious effect on coagulation dysfunction under these circumstances.

Treatment of vitamin K-dependent coagulation factor deficiency and subarachnoid hemorrhage

BACKGROUND： In adults, vitamin K-dependent coagulation factor deficiency （VKCFD） increases in the recent years. We treated a VKCFD patient with subarachnoid hemorrhage, with favorable outcomes.METHODS： A 19-year-old male student with VKCFD was treated at our hospital. The initial treatment was injection of a large dose of vitamin K and fresh plasma, and then with oral high dose of vitamin K4.RESULTS： At 4 weeks after admission, the focus of hemorrhage subsided, neurological examination was normal, and the patient was discharged.CONCLUSIONS： VKCFD is rare and its diagnosis should be based on the history of the patient and the results of laboratory examinations. A large dose of vitamin K is the fi rst choice of treatment.

Preparation and Structure of a New Coagulation Factor XI Catalytic Domain for Drug Discovery

Human blood coagulation factor XI （FXI） is a key enzyme in the amplification phase of blood coagulation cascade, and is recognized as an important target for anti-coagulant development in recent years. We designed a new mutant form of FXIa catalytic domain rhFXI370-607 （N73Q-N113Q-C123S）, and report here the facile preparation, protein crystallization, and crystal structure of this protein. We highlight a few unique structural features of FXIa after comparison with the trypsin family serine proteases at sequence and structural levels. This work provides a foundation to develop new small molecular FXIa inhibitors with increased potency and specificity.

Acquired hemophilia A in solid cancer: Two case reports and review of the literature

Acquired hemophilia A （AHA） is a rare, hemorrhagic autoimmune disease, whose pathogenesis involves reduced coagulation factor Ⅷ （FⅧ） activity related to the appearance of inhibitors against FⅧ. Common etiological factors include autoimmune diseases, ma-lignancy, and pregnancy. We report two cases of AHA in solid cancer. The first case is a 63-year-old man who developed peritoneal and intestinal bleeding after gastrectomy for gastric cancer. He was diagnosed with AHA, and was treated with prednisone, followed by cyclophosphamide. In the second case, a 68-year-old man developed a subcutaneous hemorrhage. He was diagnosed with AHA in hepatocellular carcinoma on CT imaging, and treated with rituximab alone. Hemostasis was achieved for both patients without bypassing agents as the amount of inhibitors was reduced and eradicated. However, both patients died within 1 yeardue to cancer progression. Successful treatment for AHA in solid cancer can be diffcult because treatment of the underlying malignancy is also required.

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.

Maltose-binding Protein Improving the Crystallizability of C2 Domain of Human Coagulation Factor V

Human coagulation Factor V（FV）, together with Factor Xa, assembles to prothrombinase complex on activated cell surface, which converts prothrombin into thrombin, leading to fibrin deposition. The C2 domain of FV is believed to be a primary anchor for the assembly of pro- thrombinase on the cell surface, and was proposed as a target to intervene with pathological thrombotic events. We report here the crystal structure of the C2 domain of FV fused to maltose-binding protein（MBP）. The fusion tag of MBP is critical to generate the crystal for this study. There is no strong interaction between MBP and FVC2. The overall structure of FVC2 is similar to the previous FVC2 structures, suggesting the MBP fusion does not perturb the molecular structure of FVC2. This crystal form of FVC2 can be used for future study of molecular interaction between FVC2 and its inhibitors.

Guidelines for Patients with Bleeding Disorders Undergoing Dentalveolar Surgeries

Dental practitioners must be well informed about the pathology, complications and treatment options associated with bleeding disorders patients. Prolongation of bleeding time can seriously complicate the patient’s condition during and after surgery, especially if there is iron deficiency anemia or any other condition accompanied by a decrease in hematopoiesis. For this reason, the dentist surgeon must be aware about the presence of such diseases in advance, in order to prevent the development of bleeding and its undesirable consequences promptly. Blood loss becomes apparent when blood exits through a natural opening in the body, for instance the nose & mouth. In this article, common medical bleeding situations with the potential to compromise the successful outcome of dental surgical procedures have been presented. Bleeding disorders is a disease group, which can be classified as deficiencies of coagulation factors, platelet disorders, vascular disorders, fibrinolytic defects and so on. Fragile blood vessels can cause bleeding, petechiae, bruising, etc. In most cases, vascular disease does not cause serious blood loss, with the exception of hereditary hemorrhagic telangiectasia. In this paper, we discuss the most common hereditary diseases associated with a deficiency of plasma coagulation factors VIII and IX, the disease of platelet deficiency (Glansman disease), the prophylaxis of bleeding in this kind of patients, and the effect of drugs on coagulation processes as well.

ABO Blood Groups and Their Relationship with Coagulation Factor VIII in Healthy Adults

Background: ABO blood group distribution defers with racial and geographic variations. They are related to diseases like cardiovascular diseases, cerebral thromboembolism. ABO blood group system may influence coagulation factor VIII which may increase the future risk of thrombosis. Aim: To assess the relation of ABO blood group with coagulation factor VIII in healthy adults. Material and Methods: A prospective type of analytical cross-sectional study was conducted in the Department of Physiology, Dhaka Medical College, Dhaka from July 2019 to June 2020. After obtaining ethical clearance, a total of 190 healthy adults were selected from different areas of Dhaka city based on inclusion and exclusion criteria, with ages ranging from 18 - 45 years. The subjects were interviewed and detailed history regarding personal, family, medical and drug were taken. Prior to sample collection, informed written consent was taken from the participants. Individuals of blood group A were selected as group A, blood group B as group B, blood group AB as group AB and blood group O as group O. Coagulation factor VIII was measured in the Department of Hematology and BMT Unit, Dhaka Medical College Hospital, Dhaka. Blood grouping was done in the Department of Physiology, Dhaka Medical College, Dhaka. Statistical Analysis: For statistical analysis, ONE way ANOVA followed by Bonferroni test were considered using SPSS 25.0 version. Results: In this study, blood group B was most common (33.2%). Coagulation factor VIII was significantly higher (p < 0.001) in blood group A (105.76% ± 11.82%), B (112.00% ± 15.02%), AB (109.80% ± 11.93%) than blood group O (82.00% ± 12.86%). No significant difference was observed among A, B and AB blood groups regarding coagulation factor VIII. Conclusions: It can be concluded that blood group A, B, AB individuals may have more chance of thrombosis due to significantly higher coagulation factor VIII than blood group O individuals.