OBJECTIVETo analyze clinical manifestation and genetic mutations in 8 Chinese pedigrees featuring hereditary dysfibrinogenemia.

METHODSProthrombin 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.

RESULTSAll 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.

CONCLUSIONp.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

The adsorption of proteins on the surface of glass slides is essential for construction of protein chips. Previously, we prepared a nickel-coated plate by the spin-coating method for immobilization of His-tagged proteins. In order to know whether the structural factor is responsible for the immobilization of His-tagged proteins to the nickel-coated glass slide, we executed a series of experiments. First we purified a His-tagged protein after expressing the vector in E. coli BL21 (DE3). Then we obtained the unfolding curve for the His-tagged protein by using guanidine hydrochloride. Fractions unfolded were monitored by internal fluorescence spectroscopy. The delta G(H20) for unfolding was 2.27 kcalmol +/- 0.52. Then we tested if unfolded His-tagged proteins can be adsorbed to the nickel-coated plate, comparing with Ni2+ -NTA (nitrilotriacetic acid) beads. Whereas unfolded His-tagged proteins were adsorbed to Ni2+ -NTA beads, they did not bind to the nickel-coated plate. In conclusion, a structural factor is likely to be an important factor for constructing the protein chips, when His-tagged proteins will immobilize to the nickel-coated slides.
Adsorption ; Fibrinogen ; Glass* ; Guanidine ; Immobilization ; Protein Array Analysis ; Spectrometry, Fluorescence

OBJECTIVETo investigate the patterns of changes in serum levels of of D-dimer, fibrinogen (FIB) and fibrin degradation product (FDP) during catheter-directed thrombolysis (CDT) in patients with acute lower-extremity deep venous thrombosis (DVT) and explore their clinical significance.

METHODSFrom June, 2014 to June, 2015, 50 patients with acute lower-extremity DVT received CDT. The serum concentrations of D-dimer, FIB and FDP were measured before, during and after CDT in all the subjects, with 50 healthy subjects serving as the control group.

RESULTSCompared with the control group, the patients in DVT group showed significantly increased serum levels of D-dimer (29.17±38.67 vs 0.21 ±0.27 µg/mL), FIB (3.66±0.95 vs 3.32±0.65 g/L) and FDP (76.14±131.48 vs 1.08±0.73 µg/mL) before CDT (P<0.05). Based on the effect of CDT, the patients with DVT were divided into recanalization group (n=34) and failed recanalization group (n=16), and the patients with recanalization had significantly increased serum concentration of D-dimer and FDP (P<0.05) and decreased FIB level (P<0.05) compared with those with failed recanalization at 24 h of CDT. D-dimer, FDP, and FIB showed no significant changes in the patients with failed recanalization after the procedure (P>0.05). Correlation analysis showed that serum D-dimer (r=0.66, P<0.05) and FDP (r=0.50, P<0.05) at 24 h of the procedure were positively correlated with the outcomes of CDT.

CONCLUSIONSerum levels of D-dimer, FIB and FDP are important indicators for evaluating and predicting the effectiveness of CDT in patients with acute DVT.

Acute Disease ; Blood Coagulation ; Case-Control Studies ; Catheters ; Fibrin Fibrinogen Degradation Products ; analysis ; Fibrinogen ; analysis ; Fibrinolysis ; Humans ; Thrombolytic Therapy ; Treatment Outcome ; Venous Thrombosis ; therapy

No abstract available.
Female ; Fibrin Fibrinogen Degradation Products/*analysis ; Humans ; Male ; Venous Thromboembolism/*diagnosis

No abstract available.
Female ; Fibrin Fibrinogen Degradation Products/*analysis ; Humans ; Male ; Venous Thromboembolism/*diagnosis

Background and Purpose: We investigated the relationship between the beta-fibrinogen gene (FGB) -455 G/A polymorphism and plasma fibrinogen levels in Korean ischemic stroke patients. We also determined whether the frequency of the -455 G/A polymorphism differed between two subtypes of noncardioembolic stroke: large-artery atherosclerosis (LAA) and small-vessel occlusion (SVO). Methods: A total of 267 patients with noncardioembolic stroke were enrolled. Plasma fibrinogen and other risk factors for stroke were evaluated. FGB -455 G/A genotypes were determined by polymerase chain reaction with restrictive enzyme Hae III and automatic DNA sequencing. Results: The FGB -455 G/A polymorphism was significantly associated with an elevated plasma fibrinogen level (p<0.001). The frequency of the A allele in Korean stroke patients was 16.7%. However, the frequency of the -455 G/A polymorphism did not differ between LAA and SVO. Conclusions: The plasma fibrinogen level might be affected by the -455 G/A polymorphism in noncardioembolic stroke patients. However, the LAA and SVO subtypes of ischemic stroke were not affected by the -455 G/A polymorphism.
Alleles ; Atherosclerosis ; Fibrinogen ; Genotype ; Humans ; Plasma ; Polymerase Chain Reaction ; Risk Factors ; Sequence Analysis, DNA ; Stroke

Many screening tests have been developed for the detection of FDP for the last decade. Of these, Thrombo-Wellcotest was chosen in our laboratory as a screening procedure. For the last one year, there were 121 determinations performed on 82 patients. Of the patients suspected to have DIC, 27 patients with clinical, and laboratory evidence of DIC showed the Thrombo-Wellcotest to be positive with titers ranging from 1:5 to 1:1280. Those patients without clinical or laboratory evidence of DIC gave all negative results except for 7 positives with low titers. It is our opinion that the Thrombo-Wellcotest is a simple procedure to be performed by ordinary laboratory personnel and an inexpensive test which can be afforded by most of the patients. As a whole, the Thrombo-Wellcotest is considered to be a useful screening test for the detection of FDP in serum.
Disseminated Intravascular Coagulation/diagnosis* ; Fibrin Fibrinogen Degradation Products/analysis* ; Human ; Latex Fixation Tests

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 ; genetics ; DNA Mutational Analysis ; Female ; Fibrinogen ; genetics ; Fibrinogens, Abnormal ; genetics ; Humans ; Male ; Mutation ; Pedigree

Humans ; COVID-19 ; Fibrin Fibrinogen Degradation Products ; Blood Coagulation Factors ; Survival Analysis

OBJECTIVE: To evaluate the value of conventional and age-adjusted D-dimer cut-off value combined with 2-level Wells score for diagnosis of suspected pulmonary embolism. METHODS: In the study, 335 patients with suspected pulmonary embolism who visited Peking University People's Hospital were enrolled retrospectively, then 274 patients with age over fifty years were chosen. The 2-level Wells score was applied to evaluate the clinical probability of pulmonary embolism, the diagnostic value of traditional D-dimer cut-off value (500 μg/L) and age adjusted D-dimer cut-off value (age×10 μg/L above 50 years) combined with Wells score no greater than 4 were compared. Computed tomography pulmonary arteriography (CTPA) was considered as the gold standard for diagnosis of pulmonary embolism. RESULTS: (1) The area under a receiver operating characteristic (ROC) curve (AUC) in analysis of the combination of Wells score no greater than 4 and traditional D-dimer cut-off value was 0.764 (95%CI: 0.703-0.818). On the other hand, the AUC in a ROC analysis of the combination of Wells Score no greater than 4 and age-adjusted D-dimer cut-off value was 0.814 (95%CI:0.756-0.863). These two results did not differ statistically (Z=0.05, P=0.121). (2) The sensitivity, specificity, positive predictive value, negative predictive value and Youden index of the diagnosis of pulmonary embolism of the combination of traditional D-dimer cut-off value and 2-level Wells Score were 100%, 48.9%, 28.8%, 100%, and 0.49, respectively. Meanwhile, the sensitivity, specificity, positive predictive value, negative predictive value and Youden index of the diagnosis of pulmonary embolism of the combination of age-adjusted D-dimer cut-off value and 2-level Wells Score were 97.4%, 62.3%, 35.5%, 99.1%, and 0.60, respectively. Compared with using traditional D-dimer cut-off value, using age-adjusted D-dimer cut-off value could improve the diagnosis specificity (traditional D-dimer cut-off value group: 48.9%, age-adjusted D-dimer cut-off value group: 62.3%) of pulmonary embolism without reducing the sensitivity (traditional D-dimer cut-off value group: 100%, age-adjusted D-dimer cut-off value group: 99.1%). (3) Among the 222 patients with Wells Score no greater than 4, 90 patients were with D-dimer less than traditional cut-off value (500 μg/L), and 25 patients (account for 11.3% of all 222 patients) were with D-dimer between traditional cut-off value and age-adjusted cut-off value. CONCLUSION The application of age-adjusted D-dimer cut-off value can improve the diagnostic specificity of pulmonary embolism in patients over 50 years, without reducing the sensitivity. It can be used for ruling out suspected pulmonary embolism safely.
Fibrin Fibrinogen Degradation Products/analysis* ; Humans ; Predictive Value of Tests ; Pulmonary Embolism/diagnosis* ; Retrospective Studies ; Sensitivity and Specificity