Table 1
Primer sequences, restriction enzymes used for genotyping, and length of polymerase chain reaction-restriction fragment length polymorphism products [19-22].
| Substitution | Primer Sequence (5’>3>) | Restriction Enzyme | DNA Fragment (bp) | |
|---|---|---|---|---|
| Wild Type Allele | Polymorphic Allele | |||
| GP Ia/IIa | F: GTG TTT AAC TTG AAG ACA TAT | TaqI | 92; 23 | 115 |
| c.807C>T | R: ACC TTG CAT ATT GAA TTG CTT | |||
| GP Ibα | F: GGC GAG TGT AAG GCA TCA GG | AvaII | 223; 26 | 249 |
| c.-5T>C | R: ACA CTT CAC ATG GAC TGG AT | |||
| GP Ibα | F: ACA CTT CAC ATG GAC TCC AT | – | A-520; B-480; | |
| VNTR | R: GGG TCA TTT CTG GAG CTC TC | C-440; D-400 | ||
| GP Ibα | F: GCC AGC CAC CTA GAA GTG AA | HhaI; BsaHI | 245; 170; | 286; 245; 45 |
| c.5/92C>T (HPA-2) | R: AAA AGC AAA AGG CAG GAG GT | 116; 45 | ||
| GP IIb/IIIa | F: TTC TGA TTG CTG GAC TTC TCT T | MspI | 223; 39; 6 | 173; 50; 39; 6 |
| PIA1/2 | R: TCT CTC CCC ATG GCA AAG AGT | |||
| GP VI | F: ACA TCC ACA ACA GTC CAG TG | HpaI | 120; 112; | 112; 95; 47; 25 |
| c.13254T>C | R: ATC GAG AAG TCT AGG CAG AG | 47 | ||
| vWF | F: AAG CCA GGA TTA GAA CCC GAG TCG | KpnI | 276; 406 | 682 |
| c.24/1282A>G | R: AAC TCC ATG GTT CTG GAT GTG GCG TTC | |||
| FVII | F: TCG CAT GAT TGC TAT GGG AC | EcoRI | 284; 74 | 284; 82 |
| c.-323P0/10 | R: GTT GAC ATT CCC CAT GGG AC | |||
| β-Fibrinogen | F: GAA CAT TTT ACC TTA TGT GAA TTA AGG | HindIII | 290; 194; | 379; 290 |
| c.-148C>T | R: GAA GCT CCA AGA AAC CAT CC | 185 | ||
[i] F: forward primer; R: reverse primer.
Table 2
Clinical characteristics.
| Characteristics | Patients With Thrombosis (n = 57) | Patients Without Thrombosis (n = 51) | p Value |
|---|---|---|---|
| Age: mean (SD) | 66.98 (13.42) | 60.17 (15.58) | 0.016a |
| Males: n (%) | 21 (45.7) | 25 (54.3) | 0.96b |
| Females: n (%) | 34 (54.8) | 28 (45.2) | |
| Disease duration: months: median (min-max) | 28.0 (8.0-184.0) | 30.0 (14.0-180.0) | 0.70c |
| Spleen size, cm: median (min-max) | 14.60 (4.88) | 14.18 (3.37) | 0.87c |
| Hb (g/dL): mean (SD) | 14.68 (2.99) | 15.34 (3.05) | 0.26a |
| RBC count (1012/L): median (min-max) | 5.19 (2.75-7.34) | 5.17 (3.21-8.04) | 0.87c |
| PCV (L/L): median (min-max) | 0.45 (0.22-0.63) | 0.45 (0.31-0.64) | 0.50c |
| MCV (fL): median (min-max) | 83.0 (70.0-100.0) | 87.0 (65.0-101.0) | 0.02c |
| MCH (pg): median (min-max) | 27.0 (17.0-34.0) | 28.0 (23.0-33.0) | 0.048c |
| Platelet count (109/L): median (min-max) | 553.0 (21.0-1554.0) | 581.0 (164.0-1700.0) | 0.65c |
| WBC count (109/L): mean (SD) | 9.34 (1.46) | 9.46 (1.34) | 0.71a |
| Leukocyte count (109/L): median (min-max) | 10.14 (4.58-24.67) | 10.15 (4.00-22.00) | 0.64c |
| Monocyte count (109/L)L median (min-max) | 0.60 (0.10-1.39) | 0.60 (0.20-2.68) | 0.80c |
| Basophils (109/L): median (min-max) | 0.09 (0.00-0.36) | 0.08 (0.00-0.52) | 0.62c |
| BMI: mean (SD) | 26.56 (4.47) | 27.67 (5.04) | 0.30a |
| Diabetes mellitus: n (%) | 7 (70.0) | 3 (30.0) | 0.11b |
| Smoking: n (%) | 4 (50.0) | 4 (50.0) | 0.75b |
| JAK-2: n (%) | 41 (52.6) | 37 (47.4) | 0.002b |
| CALR: n (%) | 1 (11.1) | 8 (88.9) | 0.03b |
SD: standard deviation; Hb: hemoglobin; RBC count: red blood cell count; PCV: packed cell volume; MCV: mean corpuscular volume;
MCH: mean corpuscular Hb; WBC count: white blood cell count; BMI: body mass index.
Table 3
Thrombotic complications.
| Thrombosis: n (%) | |||
|---|---|---|---|
| ET/PMF | PV | p Valuea | |
| Arterial: | 36 (73.5) | 13 (26.5) | 0.03 |
| Cardiac (myocardial infarction) | 11 | 4 | NS |
| Neurological (TIA and ischemic stroke) | 20 | 8 | NS |
| Peripheral arterial thrombosis | 5 | 1 | NS |
| Venous: | 7 (50.0) | 7 (50.0) | 0.36 |
| Deep venous thrombosis | 4 | 3 | NS |
| Pulmonary artery thrombosis | 0 | 1 | NS |
| Splanchnic vein thrombosis | 3 | 3 | NS |
ET: essential thrombocythemia; PMF: primary myelofibrosis; PV: polycythemia vera; NS: not significant; TIA: transient ischemic attack.
Table 4
Distribution of polymorphisms in arterial and venous thrombosis.
| Polymorphisms | GP IIa/IIIa PIA1/2 | GP Ia/IIa c.807C>T | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Genotypes | TT | TC | CC | CC | CT | TT | CC | CT | TT |
| Arterial n (%) | 34 (47.2) | 12 (48.0) | 2 (28.6) | 44 (46.3) | 3 (37.5) | 1 (50.0) | 13 (38.2) | 23 (42.6) | 13 (65.0) |
| Venous n (%) | 10 (13.2) | 1 (4.0) | 2 (28.6) | 12 (12.6) | 0 (0.0) | 1 (50.0) | 5 (14.7) | 7 (13.0) | 2 (10.0) |
| Polymorphisms | GP Ibα VNTR | GP Ibα c.4T>C Kozak | GP VI c.13254T>C | ||||||
| Genotypes | CC | CD | DD | TT | TC | CC | TT | TC | CC |
| Arterial n (%) | 41 (46.3) | 4 (57.1) | 4 (66.7) | 38 (48.1) | 11 (40.7) | 0 (0.0) | 43 (46.7) | 6 (37.5) | 0 (0.0) |
| Venous n (%) | 10 (10.6) | 2 (28.6) | 1 (16.7) | 11 (13.9) | 3 (11.1) | 0 (0.0) | 11 (12.0) | 3 (18.8) | 0 (0.0) |
| Polymorphisms | β-Fibrinogen c.-148C>T | FVII c.-323P0/10 | vWF c.24/1282A>G | ||||||
| Genotypes | CC | CT | TT | P0/P0 | P0/10 | P10/10 | AA | AG | GG |
| Arterial n (%) | 18 (40.0) | 30 (57.7)a | 1 (12.5) | 29 (38.2) | 9 (69.2)b | 0 (0.0) | 39 (41.1) | 0 (0.0) | 0 (0.0) |
| Arterial n (%) | 5 (11.1) | 7 (13.5) | 1 (12.5) | 8 (10.5) | 2 (15.4) | 1 (100.0) | 12 (12.6) | 0 (0.0) | 0 (0.0) |
FVII: coagulation factor VII; vWF: von Willebrand factor.
Data are presented as n (number) (%) in arterial and venous thrombosis groups. Comparisons reaching statistical significance are bold.
Table 5
Multivariate logistic regression.
| Variable | OR | p Value | 95% CI-OR |
|---|---|---|---|
| TT of GP Ia/IIa c.807C>T | 3.83 | 0.032 | 1.13-13.03 |
| CT of β-Fibrinogen c.-148C>T | 2.72 | 0.042 | 1.04-7.12 |
| c.-323P0/10 + c.-323P10/10 of FVII | 3.44 | 0.08 | 0.86-13.69 |
| Age | 1.03 | 0.041 | 1.001-1.068 |
| MCV | 0.96 | 0.003 | 0.94-0.97 |
[i] OR: odds ratio; 95% CI: 95% confidence interval; MCV: mean corpuscular volume.