The clinical value of F-18 FDG PET/CT in patients with secondary hemophagocytic syndrome / 中国实验血液学杂志

The aim of this study was to investigate the role of F-18 fluoro-2-deoxyglucose positron emission tomography/computed tomography (F-18 FDG PET/CT) in diagnosis and prognostic evaluation of secondary hemophagocytic syndrome (HPS). A total of 11 secondary HPS patients examined with 18F-FDG-PET/CT were retrospectively analyzed. The diagnostic value of F-18 FDG PET/CT for malignancy detection was assessed. The values of maximum standardized uptake value (SUV(max)) in spleen (SUVS(p)) and in bone marrow (SUVBM) were measured to analyze their relationship with various laboratorial parameters and clinical outcome of secondary HPS patients. The results showed that 4 out of the 11 patients had malignancies, the sensitivity, specificity and diagnostic accuracy of F-18 FDG PET/CT for malignancy detection were 100%, 66.7% and 75% respectively, the SUV(max) of spleen and bone marrow showed no significant correlation with laboratorial parameters, a maximum SUVS(p) of 3.10 and a maximum SUVBM of 3.47 were the optimal cutoffs for predicting patients' outcome, the increased uptake of F-18 FDG in the BM and spleen were significantly associated with shorter survival time according to univariate analysis. It is concluded that 18F-FDG PET/CT may especially play an important role in diagnosis and predicting outcome of secondary HPS for the small sample size.

Expression of c/ebpα gene in MDS and its clinical significance / 中国实验血液学杂志

This study was aimed to investigate the expression of CCAAT/enhancer binding protein alpha gene (c/ebpα) in patients with myelodysplastic syndromes (MDS) and to explore the significance of c/ebpα in pathogenesis and progression of MDS. Real time quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) method was used to detect the expression level of c/ebpα mRNA in bone marrow mononuclear cells (BMMNC) of 33 patients with MDS and 14 normal controls. The results showed that the expression level of c/ebpα mRNA in low-risk and high-risk MDS was significantly lower than that of normal controls (p < 0.01, p < 0.001, respectively), moreover, high-risk MDS showed lower c/ebpα mRNA expression compared with low-risk MDS (p < 0.05). c/ebpα mRNA expression level in MDS was not correlated with sex, age and peripheral blood cell amount, while the ratio of blast cells in bone marrow was in the c/ebpα mRNA low expression group significantly higher than that in the high expression group (p < 0.01). It is concluded that down-regulation of c/ebpα mRNA expression level has closely associated with the pathogenesis of MDS, the c/ebpα may be an important molecular biological marker of MDS; the degree of down-regulated c/ebpα has closely related to the progression of MDS.

Correlative study between JAK2 mutation and thrombosis in patients with myeloproliferative neoplasm / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the frequency and clinical implication of JAK2 mutation in patients with myeloproliferative neoplasm(MPN)and the correlation between the mutation and thrombosis.</p><p><b>METHODS</b>The clinical and laboratory data of 107 MPN patients was retrospectively analyzed. JAK2 mutation were detected with allele-specific polymerase chain reaction (AS-PCR) and sequencing. The significance of the mutation in disease diagnosis and molecular pathogenesis and the correlation between the mutation and thrombosis was analysed.</p><p><b>RESULTS</b>JAK2 mutation was detected in 71 (66.4%) and thrombosis in 34 (31.8%) of the 107 MPN patients. Thrombosis occurred in 34.8% (16/46) of polycythemia vera (PV), 32.6% (14/43) of essential thrombocythemia (ET), and 22.2% (4/18) of primany myelofibrosis (PMF) patients. The difference among the 3 groups was not significant (χ(2) = 0.96, P > 0.05). The frequency of thrombosis in JAK2(+) MPN patients (82.4%, 28/34) was higher than that in JAK2(-) MPN patients (17.6%, 6/34) (χ(2) = 5.71, P < 0.05). The frequency of thrombosis in MPN patients > 60 years was higher (41.5%, 27/65) than that in patients < 60 years (16.7%, 7/42) (χ(2) = 7.28, P < 0.01).</p><p><b>CONCLUSION</b>JAK2 V617F mutation occurs in significant percentage of Chinese patients with MPN. Patients with JAK2 mutation and older age are more succeptible to thrombosis. JAK2 mutation screening in patients with unknown thrombosis is helpful to reveal the underlying latent-MPN.</p>

Intron 1 and 22 inversions in factor VIII gene in patients with haemophilia A / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze intron 1 and 22 inversions in factor VIII (FVIII) gene in hemophilia A (HA) patients and and their families and to investigate the correlation between intron inversion and FVIII antibody.</p><p><b>METHODS</b>All patients were detected FVIII: C and FVIII antibody. In addition, 81 unrelated HA patients were directly detected by multiplex PCR and long-distance PCR for intron 1 and 22 inversions in FVIII gene. Pedigree investigation for some patients were conducted.</p><p><b>RESULTS</b>In 81 unrelated HA patients, 3 severe cases were found intron 1 inversion which accounted for 4.6% of total 65 severe cases. Of the 3 cases, one was FVIII antibody positive. Two female family members of a intron 1 inversion patient were identified as one carrier and one non-carrier. Twenty five of 65 (38.5%) severe cases were found intron 22 inversion. Of the 25 cases 1 was FVIII antibody positive. Nine female members in 5 HA families which had patients with intron 22 inversion were identified as 7 carries and 2 non-carriers.</p><p><b>CONCLUSION</b>Besides intron 22 inversion, intron 1 inversion was another important molecular defect in resulting in severe HA. Intron inversion analysis can also be used for deviation rectification of experiment grouping in HA patients. Intron 1 and 22 inversions may be one of the higher risk factors for resulting in FVIII antibodies.</p>

Clinical significance of anticoagulant proteins detection in patients with thrombotic events / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence and the risk of natural anticoagulants such as plasma protein C (PC), protein S (PS) and antithrombin (AT) deficiency in thromboembolic patients with no evident acquired factors.</p><p><b>METHODS</b>Clotting assays on French STAGO autoanalyzer were used to detect the activity of plasma PC, PS and AT in 85 patients with thrombotic disease and 50 sex and age matched healthy controls.</p><p><b>RESULTS</b>Among the 85 enrolled patients (18 arterial and 67 venous thromboembolism), male to female ratio was 1.4 and the median age was 42 years (17-69). The activity of plasma PC, PS and AT in the pre-therapy thrombotic disease group, the thrombo-recurrence group, and the age < or = 45 years group were significantly lower than that is the healthy control group, the first thrombotic episodes group and the age > 45 years group respectively (P < 0.001, P < 0.01, P < 0.01). The overall deficiency rate of these three natural anticoagulants was 30.6%, PS deficiency was the commonest (10.6%), the second was PC deficiency (8.2%), AT deficiency and combined deficiency each accounted for 5.9%.</p><p><b>CONCLUSION</b>The PC, PS and AT protein deficiencies are frequent in Chinese thromboembolic patients, they are the independent risk factors for the thrombotic events and recurrence.</p>

Identification of a novel mutation of F (13) A gene in a pedigree with factor XIII deficiency / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore F (13) A gene mutation in a pedigree with hereditary coagulation factor XIII (FXIII) deficiency.</p><p><b>METHODS</b>The FXIII deficiency was diagnosed by clot solubility test and other standard laboratory clotting tests. All exons, exon-intron boundary sequences of F(13) A gene were amplified by PCR and the products were sequenced directly. Any mutation identified by direct sequencing was confirmed by reverse sequencing. The mutation identified in the proband was screened in the family members.</p><p><b>RESULTS</b>The assays of PT, Qiulan, fibrinogen leveling, platelet counts, bleeding time were normal and the clot solubility test was positive in the proband. The homozygous deletion of 33 nucleotides (127067de133) in exon 10 of F(13) A gene which resulted in deletion of 11 amino acids in FXIIII A protein with 720aa residues was identified in the proband. Family studies showed that the mutation was inherited from the parents both of whom carried the heterozygous deletion mutation.</p><p><b>CONCLUSION</b>The homozygous 127067de133 mutation of F(13) A gene is responsible for the disorder of the pedigree.</p>

A novel mutation in antithrombin gene results in hereditary antithrombin deficiency / 中华血液学杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Immuno-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.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSION</b>This 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.</p>

Severe hereditary coagulation factor V deficiency caused by two novel heterozygous mutations / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify gene mutations of a pedigree with inherited factor V (FV) deficiency.</p><p><b>METHODS</b>The activated partial thromboplastin time (APTT), prothrombin time (PT), FV activity (FV:C) and FV antigen (FV:Ag) tests were performed for phenotypic diagnosis. The genomic DNA was extracted from the peripheral blood of the proband and all the 25 exons and their flanks of FV gene were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations were further confirmed by restriction enzyme digestion.</p><p><b>RESULTS</b>APTT, PT, TT, FV:C, FV:Ag of the proband were 249.2 s, 46.6 s, 17.9 s, 0.1% and 1.5%, respectively. FII, FVII, FVIII, FIX, FX activities, vWF and Fg were within normal ranges. Taking the GenBank Z99572 sequence as the reference, four mutations were identified in FV gene of the proband. They were a heterozygous two bases deletion in exon 13 (2238 approximately 2239delAG) introducing a frameshift and a premature stop at codon 689, and a heterozygous missense mutation in exon 23 (G6410T) resulting in the substitution of Gly for Val at codon 2079, respectively. The proband's father and mother were heterozygous for G6410T and for 2238 approximately 2239delAG, respectively.</p><p><b>CONCLUSION</b>The severe FV deficiency of the proband is caused by a frameshift mutation of 2238 approximately 2239delAG and a missense mutation of G6410T, which haven't been identified before.</p>

Fibrinogen beta chain gene mutation contributes to one congenital afibrinogenemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify the fibrinogen (Fg) gene mutations in a Chinese pedigree of congenital afibrinogenemia.</p><p><b>METHODS</b>The plasma Fg activity and protein of the proband and his family members were detected. Genomic DNA was isolated from the peripheral blood mononuclear cells. All the exons and exon-intron boundaries of fibrinogen gene were amplified by PCR and sequenced thereafter.</p><p><b>RESULTS</b>Two mutations, 7972 del G in FGB and T2543A in FGG, were found in the proband.</p><p><b>CONCLUSIONS</b>FGG2543 is a polymorphism site, which lead to the polymorphism of gamma144 I/K. The G deletion at base 7972 of FGB contributes to the frameshift mutation after amino acid 419, resulting in the truncated beta chain without the terminal 27 amino acids. The latter may contributes to the pathogenetic mechanisms in Chinese congenital afibrinogenemia patients. The G deletion at base 7972 of FGB is identified for the first time.</p>

The Examination Method of Glycoprotein CD62P and CD63 on Platelet Membrane and Their Expression in Adults Patients with Diabetes Mellitus / 中国实验血液学杂志

To study the optimal examination method of CD62P and CD63 and investigate platelet activation in patients with diabetes mellitus (DM), whole blood labeled directly with monoclonal antibodies CD62P and CD63 and flow cytometry were used to evaluate the positive percentages and the mean fluorescence intensity of CD62P and CD63. The specimens of peripheral blood obtained from 10 healthy adults were divided into two groups. In the unfixing group, the positive percentages of CD62P and CD63 at the periods of 30, 60, 90 and 120 minutes after staining were (7.57 +/- 2.33)%, (20.50 +/- 5.70)%, (28.70 +/- 5.67)% and (36.52 +/- 6.13)%, and, (0.89 +/- 0.36)%, (1.11 +/- 0.84)%, (2.35 +/- 2.02)% and (5.43 +/- 3.66)% respectively, their respective MFI were 1.57 +/- 0.13, 1.88 +/- 0.08, 2.00 +/- 0.09 and 2.38 +/- 0.22 and 3.91 +/- 0.11, 4.07 +/- 0.16, 4.38 +/- 0.14 and 4.44 +/- 0.19. However, in fixing group with 1% paraformaldehyde, the results had not any obvious change and almost were same. Besides it, the positive percentages of CD62P and CD63 in 37 adult patients with DM were (14.11 +/- 6.68)% and (2.71 +/- 1.74)%, significantly higher than that in the normal controls. It is concluded that the CD62P and CD63 on platelet membrane were very sensitive and would be easily activated in vitro, all manipulations that includes labeling with antibody, incubation and detection using flow cytometry should be finished within 30 minutes after samples collected. While fixing by using 1% paraformaldehyde can steady the labeling compounds and effectively prevent the artificial activation of platelet, and keep the stable results within two hours after the samples labeled. In adult patients with DM, the relationship between the cardiovascular complication of diabetes and platelet activation might be existed.