1.Determination of the trace levels of urinary fibrinopeptides by high-performance capillary electrophoresis.
Ming-Gang CHENG ; Yan-Hua HAO ; Jian-Hua CAO ; Tong LIANG ; Ke-Yuan ZHOU ; Guang-Xin LING
Journal of Southern Medical University 2008;28(6):1005-1007
<b>OBJECTIVEb>To establish a high-performance capillary electrophoresis (HPCE)-based method for detection of trace amount of urinary fibrinopeptide A and B (FPA and FPB, respectively) as the specific molecular markers of thrombus formation in vivo.
<b>METHODSb>The HPCE system consisted of a 25 cm x 50 microm (inner diameter) coated capillary column, 0.1 mol/L phosphoric acid buffer (pH 2.5) and a UV-detector (wavelength at 190 nm). To improve the sensitivity and reproducibility, solid-phase extraction of FPA and FPB in the urine was performed using a Sep-pak C18 column, with a synthetical fibrinopeptide B-Tyr (FPB-Tyr) as the internal standard.
<b>RESULTSb>With this HPCE method, optimal separations of FPA, FPB and FPB-Tyr was achieved within 16 min, with the migration time of 7.28 min, 14.31 min and 15.22 min, respectively. The adjusted peak area ratios of FPA or FPB and the internal standard showed good linearity with the corresponding concentrations of FPA or FPB spiked in the urine(R>0.99). Under the above chromatography conditions, the minimum detection concentration of FPA and FPB in untreated urine was 30 microg/L and 40 microg/L, respectively, and the assay precision and recovery of FPA and FPB were acceptable.
<b>CONCLUSIONb>The method we established is reliable and specific for separation and identification of fibrinopeptides and other bioactive peptides.
Electrophoresis, Capillary ; methods ; Fibrinopeptide A ; urine ; Fibrinopeptide B ; urine ; Humans ; Reproducibility of Results
2.Effect of Snake Venom: Arginine-esterase on the Fibrinolytic Activity.
Jung Min PARK ; Jae Whan LIM ; Hyung Kook PARK ; Ki Bum SUNG ; Moo Young AHN ; Hyun Kil SHIN
Journal of the Korean Neurological Association 1995;13(3):464-472
BACKGROUND AND PURPOSE: Arginine esterase(Ancrod), a thrombin-like enzyme, purified from the venoms of Agkistrodon halys, has known to cleave fibrinopeptide A from the fibrinogen and lead to circulation of soluble noncross-linked "ancrodfibrin', which stimulates endogenous T-PA release.Many authors have suggested clinical applicability of this enzyme,but clinical studies on its fibrinolytic action has been insufficient.Thus we studied the influence of this enzyme on fibrinolytic activity in cerebral infarction. METHOD: We observed the change of euglobulin fibrinolytic activity, t-PA antigen, t-PA activity, fibrinopeptide A, fibrinogen, FDP and D-dimer, during 12 hours after a bolus intravenous administration of 0.25 unit of the arginine esterase to the 9 patients with cerebral infarction. RESULT:There was no change of the euglobulin fibrinolytic activity, fibrinopeptide A and t-PA Ag but there was significant increase in both t-PA activity and FDP, D-dimer and significant decrease in fibrinogen. CONCLUSION: Our result suggest that arginine esterase converts fibrinogen to a fibrin polymer which has a increased susceptibility to lysis by plasmirl This enzyme seems to amplify T-PA activity through the consequent increase in FT)P, because there is no increase in the euglobulin fibrinolytic activity, fibr'mopeptide A and t-PA Ag suggesting direct T-PA release. Arginine esterase, having action of effective defibrinogenation and safe fibrinolysis,could be used for the thrombus related disease.
Administration, Intravenous
;
Agkistrodon
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Arginine
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Cerebral Infarction
;
Fibrin
;
Fibrinogen
;
Fibrinopeptide A
;
Humans
;
Polymers
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Snake Venoms*
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Snakes*
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Thrombosis
;
Venoms
3.Values of fibrinogen degradation products (FDPs) and D-dimmer in healthy people
Journal of Practical Medicine 2002;435(11):42-43
The study involved 32 healthy blood donors (16 men and 6 women). Plasma D-dimer and FDPs levels were determined by semi-quantitative method, using chemical procedure of the Stago Inc., France. The results showed that plasma FDPs level in healthy people was lower than 40 g/ml (93.75% of the participants had lees than 20 g/ml). Plasma D-dimer level was 1-4 g/ml, with 75% of the participants had D-dimer values ranged from 1 to 2 g/ml.
Fibrin Fibrinogen Degradation Products
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Pharmaceutical Preparations
4.The Change of Plasma Concentrations of Fibrinogen Degradation Products and Fibrin Degradation Products During the Open Heart Surgery.
Seong Cheal RYU ; Sung WOO ; Kang Hee CHO ; Yeong Gwon GOO ; Sung Sub PARK
Korean Journal of Anesthesiology 1993;26(3):499-505
There were many researches, which qualitative or quantitative assays were performed about fibrinolysis and the degree of activation of coagulation system. Authors measured fibrinogen degradation products(FbDP) and fibrin degradation products(FbDP) by monoclonal enzymeimmunoassay, instead of polyclonal method in 12 cases of cardiopulmonary bypass(CPB). 1) The increase of FgDP after sternotomy is verifying the significant fibrino(geno) lysis occured by stimulation of sternotomy. 2) By the result that FgDP was significantly increased compared with FbDP, primary fibrinogenolysis is more important phenomenon than secondary fibrinolysis during CPB. 3) FbDP and FgDP were most significantly increased immediately before the end of CPB and after CPB. 4) Increased FgDP was decreased after CPB but FbDP was still elevated 5 hours after CPB. According to the above results, CPB induced primary fibrinogenolysis and secondary fibrinolysis in open heart surgery.
Cardiopulmonary Bypass
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Fibrin Fibrinogen Degradation Products*
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Fibrin*
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Fibrinogen*
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Fibrinolysis
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Heart*
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Plasma*
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Sternotomy
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Thoracic Surgery*
5.Evaluation of an Automated Coagulation Analyzer Coapresta 2000.
Jonghyeon CHOI ; Sungwook SONG ; Yongjung PARK ; Jong Rak CHOI ; Jaewoo SONG
Laboratory Medicine Online 2011;1(2):94-99
BACKGROUND: The Coapresta 2000 (Sekisui Medical CO., LTD, Japan) is a fully automated random-access multiparameter hemostasis coagulation analyzer, which is equipped with a photo-optical clot detection unit and a cap-piercing system. It is able to perform clotting time assays as well as colorimetric assays (synthetic substrate method and latex turbidimetric method). In this study, we evaluated the analytical performance of the Coapresta 2000 for coagulation test items and compared with that of the ACL-TOP (Instrumentation Laboratory, Lexingtion, MA, USA) analyzer, which is currently used for routine coagulation test items in our hospital. METHODS: The Coapresta 2000 was evaluated with respect to its technical characteristics in the determination of 8 routine coagulation test items: prothrombin time, activated partial thromboplastin time, fibrinogen, fibrin-degradation product (FDP) antithrombin III, D-dimer, factors VIII and IX. Analyse-it (Analyse-it Software Ltd, UK) and SigmaStat (Systat Software, Inc., USA) were used for statistical analysis between items on the Coapresta 2000 and the ACL-TOP analyzer. RESULTS: The intra-assay and inter-assay coefficients of variation (CV) were below 5% for both groups of samples having values within the reference interval and outside the reference interval. Significant interference was observed with hemolytic and icteric samples. Carryover was not detected. The results obtained by Coapresta 2000 were well correlated with those obtained by the ACL-TOP analyzer (r2 in the range from 0.781 to 0.969). CONCLUSIONS: We concluded that Coapresta 2000 analyzer was well correlated with ACL-TOP analyzer for the routine coagulation test items tested.
Antithrombin III
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Fibrin Fibrinogen Degradation Products
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Fibrinogen
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Hemostasis
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Latex
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Partial Thromboplastin Time
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Prothrombin Time
6.Clinical and genetic analysis of 8 Chinese pedigrees with inherited dysfibrinogenemia.
Minghua JIANG ; Xiaoou WANG ; Kuangyi SHU ; Weiyan JIANG ; Ying HUANG ; Ying LIN ; Shanshan LI ; Yunliang HU
Chinese Journal of Medical Genetics 2014;31(2):134-139
<b>OBJECTIVEb>To analyze clinical manifestation and genetic mutations in 8 Chinese pedigrees featuring hereditary dysfibrinogenemia.
<b>METHODSb>Prothrombin time(PT), activated partial thromboplastin time(APTT), thrombin time(TT), calibration of plasma protamine sulfate against TT, fibrinogen (Fg) activity, coagulation factors II, V, VII, VIII, IX, X, XI and XII of all probands and their family members were detected with an automatic blood coagulation analyzer; D-dimer(D-D) and fibrin(ogen) degradation products(FDPs) were also dtected by automatic blood coagulation analyzer, Fg antigen were detected with an immunoturbidimetry method. Exons of fibrinogen genes FGA, FGB and FGG and flanking sequences were amplified by polymerase chain reaction(PCR) and sequenced.
<b>RESULTSb>All of the probands showed normal levels of FDPs, D-dimer(D-D) and activity of coagulation factor II,V,VII, VIII, IX,X,XI, XII. Plasma PT and APTT were normal or slightly prolonged. Prolonged TT was found in all of the probands, whilst TT was not significantly shortened by protamine sulfate. Fg antigen was within the normal range, but Fg activity was significantly decreased. The Fg antigen/activity ratio was greater than 2. One proband has carried a heterozygous variant of the FGA gene g.1233G>A(p.A α Arg35His). Four have carried a heterozygous mutation of the FGB gene g.9692A>G(p.Bβ Asn190Ser). The remaining 3 had heterozygous substitution of FGG gene g.10819G>A(p.γ Arg301His). In addition, 2 polymorphisms (p.A α Thr331Ala) and p.B β Arg478Lys) were identified in FGA and FGB genes.
<b>CONCLUSIONb>p.A α Arg35His, p.B β Asn190Ser and p. γ Arg301His are responsible for the inherited dysfibrinogenemia in the 8 Chinese pedigrees. p.B β Asn190Ser is firstly reported in China. p.B β Asn190Ser and p. γ Arg301His may be mutation hot spot in the Chinese population.
Afibrinogenemia ; blood ; genetics ; Fibrin Fibrinogen Degradation Products ; analysis ; Fibrinogen ; analysis ; genetics ; Humans ; Pedigree
7.Hypercoagulable state in patients with multiple myeloma.
Zeng-Yan LIU ; Guo-Qiang ZHANG ; Wen-Zheng YU ; Xue-Xia WANG ; Jian-Rong SUN ; Na GAO ; Hua LU
Journal of Experimental Hematology 2015;23(1):142-145
<b>OBJECTIVEb>This study was to detect the plasma thrombomodulin (TM), D-dimer and fibrinogen in patients with multiple myeloma (MM) and to analyze their relationship with morbid state, and also to investigate the relationship of the expression of coagulation factor with the ratio of myeloma cells.
<b>METHODSb>ELISA was used to detect the TM level in 45 cases of MM at different stages. The plasma level of D-dimer and fibrinogen was detected by STA automatic coagulation analyser.
<b>RESULTSb>The level of plasma TM in newly diagnosed patients was higer than that in normal control group and in platform stage group (P < 0.01; P < 0.05). There were significant differences between relapsed or refractory group and normal control group or those reached platform stage group (P < 0.05). Meanwhile, the level of plasma TM in the group of thalidomide combined with chemotherapy was higer than that in newly diagnosed patients (P < 0.05). The level of plasma D-dimer and fibrinogen of MM patients was higher than that in normal controls (P < 0.01;P < 0.05). The expression of D-Dimer in relapsed or refractory group reached the maximum. Also, the level of plasma D-Dimer in group of thalidomide combined chemotherapy was higer than in newly diagnosed patients (P < 0.05). The expression of coagulation factor did not correlate with the ratio of myeloma cells.
<b>CONCLUSIONSb>Level of plasma TM, D-Dimer and fibrinogen of MM patients is higher than that in control group. The level of plasma TM and D-Dimer can be elevated when thalidomide used, which indirectly suggested the tendency for thrombosis in MM patients.
Blood Coagulation Tests ; Fibrin Fibrinogen Degradation Products ; Fibrinogen ; Humans ; Multiple Myeloma ; Thalidomide ; Thrombomodulin ; Thrombosis
8.Levels of D-dimer, fibrinogen, and fibrinogen degradation product in patients with acute chest pain.
Fangjie ZHANG ; Xiangmin LI ; Juan ZHANG ; Aimin WANG ; Liping ZHOU ; Xiaogang LI
Journal of Central South University(Medical Sciences) 2019;44(11):1209-1213
To determine the relationship among the levels of D-dimer, fibrinogen (FIB), and fibrin degradation products (FDP) in acute fatal chest pain patients.
Methods: We retrospectively analyzed the patients with aortic dissection (AD), pulmonary embolism (PE) or acute myocardial infarction (AMI) from May 1, 2017 to April 30, 2018. All the patients had a chest and/or back pain. Levels of D-dimer, FIB, and FDP were examined at the time of admission, and the patients were further diagnosed by computed tomography angiography (CTA) or percutaneous transluminal coronary intervention (PCI). The levels and negative rates of D-dimer, FIB, and FDP in patients with AD, PE, and AMI were compared.
Results: A total of 234 patients were enrolled, including 95 AD, 98 AMI, and 41 PE. In the AD group, the AMI group and the PE group, the negative ratios of D-dimer were 13.68%, 70.41% and 4.88%, respectively; the negative ratios of FDP were 24.21%, 81.63% and 24.39%, respectively. There was no significant difference in negative rates of D-dimer and FDP between the AD group and the PE group (all P>0.05), but negative rates of D-dimer and FDP were significantly higher in the AMI group than those in the AD group and the PE group (all P<0.001). The level of D-dimer in the AMI group was significantly lower than that in the AD group or in the PE group (both P<0.001), while there was no statistically significant difference between the AD group and the PE group (P>0.05). However, there were no significant difference in the FIB levels among 3 groups (all P>0.05). The FDP level in the AMI group was significantly lower than that in the AD group or in the PE group (both P<0.001), while there was no statistically significant difference between the AD group and the PE group (P>0.05).
Conclusion: The levels of D-dimer and FDP are increased in AD and PE patients and may be as the useful biomarkers for the high-risk chest pain patients but not for AMI.
Chest Pain
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Fibrin Fibrinogen Degradation Products
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Fibrinogen
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Humans
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Percutaneous Coronary Intervention
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Pulmonary Embolism
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Retrospective Studies
9.Mutation analysis of a FGG gene causing hereditary abnormal fibrinogen.
Liya JIANG ; Qiaohong ZHANG ; Wanping XU ; Yongjun ZHANG
Chinese Journal of Medical Genetics 2018;35(6):812-814
OBJECTIVE:
To study the clinical phenotype and gene mutation analysis of a hereditary abnormal fibrinogenemia family and explore its molecular pathogenesis.
METHODS:
The STA-R automatic hemagglutination analyzer to detect the proband and its family members (3 generations of 5 people) of prothrombin time(PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen activity (Fg: C), D-dimer (D-D), fibrinogen and fibrin degradation products (FDPs), plasminogen activity (PLG: A); The plasma levels of Fg: C and fibrinogen (Fg: Ag) were measured by Clauss method and immunoturbidimetry respectively. All exons and flanking sequences of FGA, FGB and FGG genes of fibrinogen were amplified by PCR, and the PCR products were purified and sequenced for gene analysis. The model was analyzed by Swiss software.
RESULTS:
The PT and APTT of the proband, her mother and sister were slightly prolonged, TT was significantly extend, Fg: C decreased significantly, Fg: Ag, PLG: A, D-D and FDPs are within the normal range; Her brother and daughter of the results are normal. Genetic analysis showed that g.7476 G>A heterozygous missense mutation in exon 8 of FGG gene resulted in mutations in arginine at position 275 of fibrinogen gamma D domain to histidine (Arg275His). Her mother and sister have the same Arg275His heterozygous mutation, brother and daughter for the normal wild type.
CONCLUSION
The heterozygous missense mutation of FGG gene Arg275His in patients with hereditary dysfibrinogenemia is associated with a decrease in plasma fibrinogen activity.
Afibrinogenemia
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genetics
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DNA Mutational Analysis
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Female
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Fibrinogen
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genetics
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Fibrinogens, Abnormal
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genetics
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Humans
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Male
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Mutation
;
Pedigree
10.D-dimer as a Prognostic Tool in Patients with Normotensive Pulmonary Embolism.
Jae Chol YOON ; Won Young KIM ; Sang Sik CHOI ; Sang Ku JUNG ; Chang Hwan SOHN ; Won KIM ; Kyoung Soo LIM ; Tae O JEONG ; Young Ho JIN ; Jae Baek LEE
Tuberculosis and Respiratory Diseases 2010;68(2):87-92
BACKGROUND: D-dimer testing is widely applied as a first step in the diagnostic work-up of pulmonary embolism (PE). Although this is the most sensitive assay for ruling out PE, the prognostic implications of D-dimer testing in patients with normotensive PE are not well known. The aim of this study was to determine if D-dimer testing on admission predicts major adverse cardiac events (MACE) in patients with normotensive PE. METHODS: A total of 180 consecutive patients with normotensive PE admitted between January 2003 and June 2009 were included. The group was divided into quartiles on the basis of their D-dimer levels. We compared the frequency of MACE by quartile of D-dimer level and estimated sensitivity, specificity, and predictive values for MACE in the first and fourth quartile. RESULTS: In the 37 (20.6%) patients with MACEs, the median D-dimer level (7.94 [IQR: 4.03~18.17] microgram/mL) was higher than in patients with a benign course (5.29 [IQR: 2.60~11.52] microgram/mL, p<0.01). The occurrence of MACEs was increased with increasing D-dimer level (p=0.017). In the first quartile (D-Dimer <2.76 microgram/mL) sensitivity, specificity, and positive and negative predictive values for predicting MACEs were, respectively, 91.9%, 29.4%, 25.2%, and 93.3%. CONCLUSION: Patients with D-dimer levels below 2.76 microgram/mL have a low risk of MACEs. Our study suggest that D-dimer level may be used to identify low risk patients with normotensive PE.
Fibrin Fibrinogen Degradation Products
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Humans
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Prognosis
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Pulmonary Embolism
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Resin Cements