OBJECTIVE: To analyze the phenotype and genotype of a patient affected with inherited antithrombin deficiency. METHODS: All exons and exon-intron boundaries of the AT genes were subjected to PCR amplification and Sanger sequencing. The influence of variants on the disease was predicted using bioinformatic software (MutationTaster). RESULTS: The results of all coagulation tests were normal, though the antithrombin activity and antigen content of the proband and his father have decreased significantly (34%, 48% and 12.97 mg/dL, 15.60 mg/dL, respectively). His mother was normal. Genetic analysis revealed that the proband and his father both carried a heterozygous g.2736dupT variant of the AT gene. Bioinformatic analysis suggested that the variant may be pathogenic. CONCLUSION The proband and his father both had type I hereditary antithrombin deficiency caused by a g.2736dupT variant of the AT gene. The variant was unreported previously.
Antithrombin III/genetics* ; Antithrombin III Deficiency/genetics* ; DNA Mutational Analysis ; Genetic Testing ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree

OBJECTIVEDynamic changes of quality and growth of Whitmania pigra were investigated to provide theoretical basis for quality control and determination of optimal harvest time.

METHODThe contents of moisture, ethanol-soluble extractive, total ash, and acid-insoluble ash, as well as antithrombin activity were determined according to Chinese pharmacopoeia (2005 edition).

RESULTQualities of W. pigra collected from different growth stages met the standards of Chinese pharmacopoeia (2005 edition). The highest amount of ethanol-soluble extract was found in 9-month-old, W. pigra, followed by 6-month-aged ones. And there was significant difference between 9-month-old and 4-month-old, 7 month-old and 11 month-old (P < 0.05). The highest antithrombin activity was detected in 6 month-old W. pigra, followed by 10 month-old. Significant differences of antithrombin activity were found between 6-month-old and 4 month-old ones, and 5 month-old (P < 0.05), however, there were no significant difference between 6 month-old ones with other samples. Eleven-month-old W. pigra got the most dry weight, and there were significant difference between 11 month-old ones with other samples (P < 0.05).

CONCLUSIONNovember, namely for 6 month old artificial cultivated W. pigra was the best harvest time in Jiangsu and Zhejiang provinces.

Animals ; Antithrombin III ; Antithrombins ; analysis ; China ; Leeches ; chemistry ; growth & development ; Medicine, Chinese Traditional ; Peptide Fragments

OBJECTIVETo investigate the antithrombin (AT) activity (AT: A) and AT antigen (AT: Ag) level in a Chinese family with type I antithrombin (AT) deficiency, and to explore the molecular mechanism of AT deficiency.

METHODSImmuno-nephelometry and chromogenic assay were used to detect the plasma level of AT: A and AT: Ag, respectively. Genomic DNA was isolated from the peripheral blood, and all the seven exons and exon-intron boundaries of AT gene were amplified by PCR and direct sequencing.

RESULTSThe plasma levels of AT: A and AT: Ag of the proband were 45% and 97 mg/L, respectively, which led to a type I AT deficiency. A heterozygous T to A mutation was found at nucleotide 9833 in exon 5 resulting in a Tyr363Stop nonsense mutation. The sequencing results from the pedigree indicated that four other members also had this mutation.

CONCLUSIONThis heterozygous nonsense mutation of T9833A in exon 5 resulting in venous thrombosis is a novel genetic defect of hereditary AT deficiency, which has not been described before.

Antithrombin III Deficiency ; genetics ; Antithrombins ; genetics ; Blood Coagulation Tests ; Female ; Humans ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVE: To investigate the correlation of procalcitonin (PCT), interleukin-6 (IL-6) and antithrombin III (AT III) with the severity of sepsis, and to compare the predictive value of the above indicators alone or in combination. METHODS: A retrospective cohort study was conducted. Eighty-five patients with sepsis admitted to the department of intensive care medicine of Shandong Provincial Hospital Affiliated to Shandong First Medical University from April 2021 to September 2022 were enrolled. General information, sequential organ failure assessment (SOFA) score and acute physiology and chronic health evaluation II (APACHE II) score within 24 hours of admission, inflammatory indicators [PCT, IL-6, serum amyloid A (SAA), neutrophil to lymphocyte ratio (NLR), and C-reactive protein (CRP)] and coagulation indicators (D-dimer and AT III) levels at admission, and 28-day prognosis were collected. The differences of the above indicators were compared among patients with different prognosis at 28 days and different severity of sepsis. The correlation between PCT, IL-6, AT III and the severity of sepsis was analyzed by Spearman rank correlation method. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of PCT, IL-6 and AT III alone or in combination on the 28-day death of patients with sepsis. RESULTS: Eighty-five patients were enrolled finally, 67 cases survived and 18 cases died at 28 days. The mortality was 21.2%. There were no statistical significant differences in gender, age and other general data between the two groups. The patients in the death group were more serious than those in the survival group, and PCT, IL-6, and CRP levels were significantly higher than those in the survival group [PCT (μg/L): 4.34 (1.99, 14.42) vs. 1.17 (0.31, 3.94), IL-6 (ng/L): 332.40 (50.08, 590.18) vs. 61.95 (31.64, 194.20), CRP (mg/L): 149.28 (75.34, 218.60) vs. 83.23 (48.22, 174.96), all P < 0.05], and AT III activity was significantly lower than that in the survival group [(53.67±28.57)% vs. (80.96±24.18)%, P < 0.01]. However, there were no significant differences in D-dimer, NLR and SAA between the two groups. Among the 85 patients, 36 had sepsis with single organ dysfunction, 29 had sepsis with multiple organ dysfunction, and 20 had septic shock with multiple organ dysfunction. With the increase of the severity of sepsis, PCT and IL-6 levels gradually increased [PCT (μg/L): 0.36 (0.19, 1.10), 3.00 (1.22, 9.94), 4.34 (2.18, 8.86); IL-6 (ng/L): 43.99 (20.73, 111.13), 100.00 (45.37, 273.00), 332.40 (124.4, 693.65)], and the activity of AT III decreased gradually [(89.81±21.42)%, (71.97±24.88)%, and (53.50±25.41)%], all with statistically significant differences (all P < 0.01). Spearman rank correlation analysis showed that PCT and IL-6 levels in sepsis patients were significantly positively correlated with the severity of the disease (r values were 0.562 and 0.517, respectively, both P < 0.01), and AT III activity was significantly negatively correlated with the severity of the disease (r = -0.523, P < 0.01). ROC curve analysis showed that PCT, IL-6, and AT III alone or in combination had some predictive value for the death of sepsis patients at 28 days. The area under the ROC curve (AUC) of the above three indicators in combination was higher than that of the individual tests (0.818 vs. 0.722, 0.725, and 0.770), with a sensitivity of 83.3% and a specificity of 73.1%. CONCLUSIONS PCT, IL-6, and AT III were significantly correlated with the severity of sepsis patients. The combined assay of the above three indicators can effectively improve the prediction of the prognosis of sepsis patients.
Humans ; Procalcitonin ; Interleukin-6 ; Antithrombin III ; Retrospective Studies ; Multiple Organ Failure ; ROC Curve ; Sepsis/diagnosis* ; Prognosis ; C-Reactive Protein/analysis* ; Anticoagulants

BACKGROUND: Effective treatment and monitoring of tuberculosis (TB) requires biomarkers that can be easily evaluated in blood samples. The aim of this study was to analyze the serum proteome of patients with TB and to identify protein biomarkers for TB. METHODS: Serum samples from 26 TB patients and 31 controls were analyzed by using nano-flow ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry in data-independent mode, and protein and peptide amounts were calculated by using a label-free quantitative approach. The generated data were analyzed by using principal component analysis and partial least squares discriminant analysis, a multivariate statistical method. RESULTS: Of more than 500 proteins identified, alpha-1-antitrypsin was the most discriminative, which was 4.4 times higher in TB patients than in controls. Peptides from alpha-1-antitrypsin and antithrombin III increased in TB patients and showed a high variable importance in the projection scores and coefficient in partial least square discriminant analysis. CONCLUSIONS: Sera from patients with TB had higher alpha-1-antitrypsin levels than sera from control participants. Alpha-1-antitrypsin levels may aid in the diagnosis of TB.
Adult ; Aged ; Antithrombin III/analysis ; Biological Markers/blood ; Chromatography, High Pressure Liquid ; Discriminant Analysis ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Proteome/*analysis ; *Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tuberculosis/*blood/genetics/metabolism ; alpha 1-Antitrypsin/analysis

BACKGROUND: The safety of plasma derivatives has been reinforced since 1980s by variable pathogen inactivation or elimination techniques. Nucleic acid amplification test (NAT) for the source plasma has also been implemented worldwide. Recently nanofiltration has been used in some country for ensuring safety of plasma derivatives to eliminate non-enveloped viruses such as parvovirus B19 (B19V) and hepatitis A virus (HAV). We evaluated the efficacy of nanofiltration for the elimination of B19V and HAV. METHODS: To verify the efficacy of nanofiltration, we adopted a 20 nm Viresolve NFP (Millipore, USA) in the scaling down (1:1,370) model of the antithrombin III production. As virus stock solutions, we used B19V reactive plasma and porcine parvovirus (PPV) and HAV obtained from cell culture. And 50% tissue culture infectious dose was consumed as infectious dose. The methods used to evaluate the virus-elimination efficacy were reverse-transcriptase polymerase chain reaction for B19V and the cytopathic effect calculation after filtration for PPV and HAV. RESULTS: B19V was not detected by RT-PCR in the filtered antithrombin III solutions with initial viral load of 6.42x10(5) IU/mL and 1.42x10(5) IU/mL before filtration. The virus-elimination efficacy of nanofiltration for PPV and HAV were > or =10(3.32) and > or =10(3.31), respectively. CONCLUSIONS: Nanofiltration would be an effective method for the elimination of B19V and HAV. It may be used as a substitute for NAT screening of these viruses in source plasma to ensure safety of plasma derivatives in Korea.
Antithrombin III/isolation & purification ; DNA, Viral/analysis ; Filtration/*methods ; Hepatitis A virus/genetics/*isolation & purification ; Humans ; Nanotechnology/*methods ; Parvovirus B19, Human/genetics/*isolation & purification ; RNA, Viral/analysis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo identify antithrombin III (AT-III) gene mutation and polymorphisms in pregnant women and parturients with cerebral venous thrombosis (CVT) using denaturing high-performance liquid chromatography (DHPLC).

METHODSThe genomic DNA was extracted from the blood samples of 50 pregnant women and parturients with CVT and 52 matched healthy women for molecular analysis using a PCR/DHPLC assay followed by DNA sequence analysis. Ten primer pairs were designed for amplifying the AT- III promoter region and exons 1-6 including the exon/intron boundaries. A rapid screening assay based on DHPLC was established to screen the mutation and polymorphisms of AT- III gene.

RESULTSSix abnormal peaks were detected in 40 of the patients by DHPLC. Direct DNA sequencing was performed on representative samples detected by DHPLC profiling. One pathogenic heterozygous G13328A missense mutation in exon 6, and a novel silent mutation in exon 4+243 G>A were identified. Six single nucleotide polymorphism (SNP) sites were found, including 4 previously reported ones in the SNP library and two were novel SNP sites. An abnormal peak was detected in the control group by DHPLC.

CONCLUSIONDHPLC allows automated and rapid high-throughput detection of AT- III gene mutation and polymorphisms in the clinical setting and prenatal diagnosis. Our findings suggested that AT- III gene mutation, as well as its polymorphisms, contributes to the occurrence of CVT in pregnant women and parturients.

Adult ; Antithrombin III ; genetics ; Base Sequence ; Case-Control Studies ; Chromatography, High Pressure Liquid ; methods ; DNA Mutational Analysis ; Female ; Genetic Testing ; methods ; Humans ; Intracranial Thrombosis ; genetics ; Molecular Sequence Data ; Mutation ; Polymorphism, Genetic ; genetics ; Pregnancy ; Pregnancy Complications, Hematologic ; genetics ; Young Adult

OBJECTIVETo investigate the pathogenesis of thromboembolism in patients with mitral stenosis in a pre-thrombotic state.

METHODSThe biochemical markers' levels in plasma for platelet activity [soluble P-selectin (GMP-140)], states of thrombin generation [antithrombin III (AT III) and protein C (PC)], fibrinolysis [D-dimer (DD), plasminogen activator inhibitor 1 (PAI-1), tissue plasminogen activator (t-PA) and FDP] and von Willebrand factor (vWF) were determined from blood specimens obtained from the femoral veins and arteries and the right and left atria of 43 consecutive patients (20 with atrial fibrillation and 23 with sinus rhythm) with mitral stenosis (MS), undergoing percutaneous mitral valvuloplasty. The same parameters were compared with those of 15 control subjects, who had no detectable heart disease, but with paroxysmal supraventricular tachycardia undergoing radiofrequency catheter ablation of the left accessory pathway through a transseptal passage.

RESULTSBlood from the left atrium contained an excessive amount of platelet activity, thrombin generation and fibrinolysis compared with the blood from the right atrium, and the femoral veins and arteries. However blood from the right atrium was much lower in these activities when compared with those from the left atrium, and the femoral veins and arteries in both groups. Compared with those in the control subjects, GMP-140 in the left atrium was significantly higher (P < 0.05) and AT III was significantly lower (P < 0.05) in patients with MS. Compared with the patients with MS and spontaneous left atrial echocontrast (LASEC) /= 2 had significantly higher levels of GMP-140 in plasma (P < 0.05), and significantly lower levels of AT III (P < 0.05) and PC (P < 0.01) levels in the left atrium. However, there were no significant differences between patients with atrial fibrillation and those with sinus rhythm regarding amounts of plasma coagulation markers in the left atrium. Univariate regression analysis revealed that LASEC was negatively correlated with plasma levels of blood from the left atria in the patients with MS.

CONCLUSIONCoagulability is increased in the left atria of patients with MS and is positively correlated with LASEC.

Adult ; Antithrombin III ; analysis ; Female ; Fibrin Fibrinogen Degradation Products ; analysis ; Heart Atria ; chemistry ; Humans ; Male ; Mitral Valve Stenosis ; complications ; P-Selectin ; blood ; Plasminogen Activator Inhibitor 1 ; blood ; Protein C ; analysis ; Regression Analysis ; Thromboembolism ; etiology ; Thrombophilia ; blood ; complications ; von Willebrand Factor ; analysis

OBJECTIVETo explore the effect and mechanism of Tongguan capsule (TGC) on coagulant and fibrinolysis system in patients with coronary heart disease (CHD) after percutaneous coronary intervention (PCI).

METHODSAdopting the prospective, randomized controlled method to observe the coagulant-fibrinolysis related indexes, including tissue plasminogen activator (t-PA), plasminogen activator inhibitor 1 (PAI-1), von Willebrand factor (vWF), antithrombin II (AT-III) and fibrinogen (FIB), in CHD patients after PCI, there were 26 patients in the treated group treated by TGC with Western medicine and 26 in the control group treated by Western medicine alone.

RESULTSAfter treatment, all the indexes were improved in both groups (P < 0.05). As comparison between the two groups, AT-III and t-PA were higher in the treated group than those in the control group; PAI-1 levels showed insignificant difference at 1 month after treatment, but at 3 months after treatment, it was significantly lower in the treated group (P < 0.05); vWF showed no significant difference either at 1 or at 3 months after treatment; and FIB was lower in the treated group both at 1 and 3 months after treatment (P < 0.01).

CONCLUSIONTGC could improve the hypercoagulant status and adjust the balance between coagulant-fibrinolysis system of CHD patients after PCI by increasing AT-III and t-PA levels, and lowering FIB and PAI-1 levels in the body.

Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Antithrombin III ; analysis ; Capsules ; Coronary Disease ; blood ; drug therapy ; therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fibrinolysis ; Humans ; Male ; Middle Aged ; Phytotherapy ; Plasminogen Activator Inhibitor 1 ; blood ; Prospective Studies ; Stents ; Tissue Plasminogen Activator ; blood ; von Willebrand Factor ; analysis