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

OBJECTIVETo observe changes in plasma thrombomodulin (TM) and D-dimer (DD) levels in neonates with sepsis, and to investigate their significance in evaluating the patients' condition and prognosis.

METHODSFifty-six neonates with sepsis were classified into extremely critical (n=13), critical (n=22) and non-critical groups (n=21) based on neonatal critical illness score (NCIS). Fasting venous blood samples were collected on admission and in the recovery phase. Plasma TM and D-dimer levels were measured using enzyme-linked immunosorbent assay and immune turbidimetry, respectively. Twenty-six healthy neonates were selected as the control group. Plasma TM and D-dimer levels were compared between groups, and the changes after treatment were determined.

RESULTSPlasma TM levels in the extremely critical, critical and non-critical groups were 25.5±6.6, 17.3±4.7 and 13.3±2.8 µg/L respectively, significantly higher than in the control group (9.8±2.7 µg/L) (P<0.01). Plasma D-dimer levels in the extremely critical and critical groups were 744±262 and 436±147 µg/L respectively, also significantly higher than in the control group (205±61 µg/L) (P<0.01). The extremely critical group had significantly higher plasma TM and DD levels than the critical group (P<0.05), and the critical group had significantly higher plasma TM and DD levels than the non-critical group (P<0.05). All patients showed significant decreases in plasma TM and DD levels in the recovery phase after treatment (P<0.01). Plasma TM and DD levels were significantly negatively correlated with NCIS (r=-0.428, P<0.01; r=-0.363, P<0.01).

CONCLUSIONSDetermination of plasma TM and DD levels may be helpful in evaluating severity and prognosis in neonates with sepsis.

Female ; Fibrin Fibrinogen Degradation Products ; analysis ; Humans ; Infant, Newborn ; Male ; Protein Multimerization ; Sepsis ; blood ; Thrombomodulin ; blood

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

The selective removal of low density lipoprotein (LDL) and fibrinogen (Fib) by degraded carrageenan was studied by the present authors. Degraded carrageenan was prepared by acid with carrageenan as the main material. The effects of acid conditions on the molecular weight were investigated, and the proper reaction conditions were ascertained. The results of infrared spectrometry indicated that the degraded carrageenan is a heparin-like polysaccharide. Then the selective removal of LDL/Fibrinogen by degraded carrageenan was studied. When molecular weight was about 10,000, pH was 5.10 and the concentration of degraded carrageenan was 800 mg/L, the average reduction percentages were 60.0% for total cholesterol(TC), 79.4% for LDL and very low-density lipoprotein (VLDL), and 93.8% for fibrinogen. There were no significant changes with relation to the level of high-density lipoprotein (HDL) and total protein (TP). So, degraded carrageenan was shown to be of good selectivity on plasma LDL/Fibrinogen apheresis.
Carrageenan ; chemistry ; Fibrinogen ; analysis ; isolation & purification ; Humans ; Hyperlipidemias ; blood ; Lipoproteins, LDL ; blood ; isolation & purification

OBJECTIVETo detect the plasma activity of von Willebrand factor-cleaving protease (ADAMTS13) in the patients with prothrombotic status, and explore the effect and significance of ADAMTS13 in the prothrombotic status. The correlation of ADAMTS13 with von Willebrand factor (vWF), thrombospondin 1 (TSP1), C-reactive protein etc, and blood pressure was simultaneously analyzed．

METHODSThe activity of ADAMTS13 in patient groups (atherosclerosis, diabetes, acute promyelocytic leukemia, cancer and sepsis, a total of 260 cases) and in control group 50 cases were evaluated by residue collagen binding assay（R-CBA）, the protein levels of TSP1 and vWF were measured by ELISA kits； the correlation of ADAMTS13 activity with CRP, creatinine, and blood pressure was analyzed with statistical soft ware.

RESULTSThe activity of plasma ADAMTS13 in patient group was significantly lower than that in normal control group(P<0.05). And the protein levels of TSP1 and vWF in the patients with prothrombotic status were higher than those in the normal controls(P<0.05). Analysis of the correlation showed that the ADAMTS13 activity correlated negatively with the levels of TSP1 protein, blood sugar, blood pressure, D-dimer, creatinine,and CRP levels (P<0.05), however, the ADAMTS13 activity did not significantly correlate with the levels of serum lipids, blood type, platelet number and hemoglobin level(P>0.05).

CONCLUSIONThe plasma ADAMTS13 activity is decreased in the patients with prothrombotic status, suggesting that the decreased ADAMTS13 activity may participate in the occurrence of prothrombotic status, and the dectection of plasma ADAMTS13 activity may help the diagnosis of pro-thrombotic disease.

ADAMTS13 Protein ; Factor Analysis, Statistical ; Fibrin Fibrinogen Degradation Products ; Humans ; Sepsis ; Thrombosis ; von Willebrand Factor