Effective Management of Polycythemia Vera With Ropeginterferon Alfa-2b Treatment.

Background: Polycythemia vera (PV) is a myeloproliferative neoplasm. Ropeginterferon alfa-2b is a new-generation polyethylene glycol-conjugated proline-interferon. It is approved for the treatment of PV at a starting dose of 100 µg (50 µg for patients receiving hydroxyurea (HU)) and dose titrations up to 500 µg by 50 µg increments. The study was aimed at assessing its efficacy and safety at a higher starting dose and simpler intra-patient dose escalation. Methods: Forty-nine patients with PV having HU intolerance from major hospitals in China were treated biweekly with an initial dose of 250 µg, followed by 350 µg and 500 µg thereafter if tolerated. Complete hematological response (CHR) was assessed every 12 weeks based on the European LeukemiaNet criteria. The primary endpoint was the CHR rate at week 24. The secondary endpoints included CHR rates at weeks 12, 36 and 52, changes of JAK2V617F allelic burden, time to first CHR, and safety assessments. Results: The CHR rates were 61.2%, 69.4% and 71.4% at weeks 24, 36, and 52, respectively. Mean allele burden of the driver mutation JAK2V617F declined from 58.5% at baseline to 30.1% at 52 weeks. Both CHR and JAK2V617F allele burden reduction showed consistent increases over the 52 weeks of the treatment. Twenty-nine patients (63.0%) achieved partial molecular response (PMR) and two achieved complete molecular response (CMR). The time to CHR was rapid and median time was 5.6 months according to central lab results. The CHRs were durable and median CHR duration time was not reached at week 52. Mean spleen index reduced from 55.6 cm2 at baseline to 50.2 cm2 at week 52. Adverse events (AEs) were mostly mild or moderate. Most common AEs were reversible alanine aminotransferase and aspartate aminotransferase increases, which were not associated with significant elevations in bilirubin levels or jaundice. There were no grade 4 or 5 AEs. Grade 3 AEs were reversible and manageable. Only one AE led to discontinuation. No incidence of thromboembolic events was observed. Conclusion: The 250-350-500 µg dosing regimen was well tolerated and effectively induced CHR and MR and managed spleen size increase. Our findings demonstrate that ropeginterferon alfa-2b at this dosing regimen can provide an effective management of PV and support using this dosing regimen as a treatment option.

[Clinical efficacies and safety of HAG regimen for patients with high-risk myelodysplastic syndromes: a multicentre study].

OBJECTIVE: To evaluate the efficacies and toxicity of HAG (HHT + Ara-C + G-CSF) regimen in patients with high-risk myelodysplastic syndromes (MDS). METHODS: A total of 97 patients with high-risk MDS received HAG regimen as the induction therapy. RESULTS: The complete remission (CR) rate of all the patients was 52.3% (45/86). The overall response (OR) rate was 66.3% (57/86). The early mortality rate was 9.3% (9/97). There was no significant difference in CR rate and OR rate between the patients aged ≥ 60 and those < 60. The OR rate was 29/34, 9/12 and 6/13 in patients with favorable karyotype, intermediate karyotype and unfavorable karyotype respectively. The OR rate was higher in patients with favorable karyotype than those with unfavorable karyotype (P = 0.038). The major adverse effect was infection. CONCLUSION: HAG regimen provides higher CR rate and OR rate for patients with high-risk MDS.

Significance of transmission electron microscopy in subtyping of monocytic leukemia.

The objective of this paper is to produce an ultrastructural classification of acute monocytic leukemia (AML-M5) in relation to clinical behaviors. The ultrastructural characteristics of blasts of the monocytic series were analyzed in 72 M5 patients by transmission electron microscopy (TEM), in terms of their content of typical monoblasts, atypical monoblasts, atypical promonocytes, and typical promonocytes in bone-marrow aspirates. Four kinds of monocytic blasts were identified by cell size and shape, nuclear profile, nucleocytoplasmic ratio, heterochromatin content, nucleolus, granules, vesicles, and Golgi apparatus. Their characteristics of remission rate, cytochemistry, immunophenotype, and cytogenetics were also investigated. The data obtained permitted M5 patients to be divided into monoblast and promonocyte types. Monoblast-type patients expressed weaker monocytic enzymograms and specific antigen staining for CD14 and CD64, compared with promonocyte-type patients. Monoblast patients had higher CR than promonocyte patients. Therefore, TEM subclassification of patients differs from and improves upon the light microscopical criteria for distinguishing monoblasts and promonocytes and has clinical significance.

[Polymorphisms of drug-metabolizing enzymes genes in a Han Chinese population].

OBJECTIVE: To survey the frequencies and characteristics of polymorphism of seven genes coding drug-metabolizing enzymes in a Han Chinese population. METHODS: Genomic DNA was extracted from peripheral blood in 1382 unrelated Han Chinese volunteers. Genotypes of CYP450, NQO1, MPO, MTHFR, NAT2, TPMT*3B(G460A) and TPMT*3C(A719G) were analyzed by PCR-RFLP. GSTM1 and GSTT1 genotypes were detected by multiple PCR and TPMT*2(G238C) genotypes by allele-specific PCR. RESULTS: The allele frequencies of wild-type, heterozygosity and homozygosity were 99.8%, 0.2% and 0 in CYP3A4*1B genotypes, 8.4%, 34.3% and 57.3% in CYP3A5*3 genotypes, 28.7%, 49.7% and 21.6% in NQO1(C609T) genotypes, 75.0%, 23.2% and 1.8% in MPO(G463A) genotypes, 25.9%, 44.9% and 29.2% in MTHFR(C677T) genotypes, 67.3%, 30.4% and 2.3% in MTHFR(A1298C) genotypes, and 96.8% 3.2% and 0 in TPMT*3C(A719G) genotypes, respectively. The frequencies of present and null genotypes were 36.1% and 63.9% in GSTM1, 54.4% and 45.6% in GSTT1 respectively. The frequencies of NAT2*4/*4, *4/*5, *4/*6, *4/*7, *5/*5, *5/*6, *5/*7, *6/*6, *6/*7 and *7/*7 genotypes were 34.5%, 4.3%, 24.3%, 18.2%, 0.1%, 1.8%, 1.5%, 5.0%, 7.6% and 2.6% respectively. The frequencies of specific NAT2 alleles were 57.9%, 3.9%, 21.8% and 16.3% for NAT2*4, *5, *6 and *7 respectively. The frequencies of genophenotypes were 81.4% and 18.6% in NAT2 fast acetylator and low acetylator respectively. The allele frequency of wild type were 100% in both TPMT*2(G238C) genotypes and TPMT*3B(G460A). The variant alleles of TPMT*2(G238C) and TPMT*3B(G460A) were not found in 1382 Han Chinese subjects. CONCLUSION: Significant differences in the distributions and frequencies in genetic polymorphisms of drug-metabolizing enzymes are identified between Han Chinese population and Caucasians, as well between Han Chinese population and Melanoderma while only some heterogeneity has been observed between Han Chinese population and other Xanthoderms in Asia.

[Polymorphisms of RAD51(G135C) and XRCC3(C241T) genes and correlations thereof with prognosis and clinical outcomes of acute myeloid leukemia].

OBJECTIVE: To investigate the impacts of RAD51G(135C) and XRCC3(C241T) genotypes on the response, adverse effects, and prognosis of acute myelocytic leukemia (AML). METHODS: RAD51(G135C), XRCC3(C241T), GSTT1, and GSTM1 genotypes were analyzed in 372 patients with AML, 226 males and 146 females, by PCR-RFLP or PCR. The Complete remission (CR) rate, adverse effects, overall survival (OS), and relapse-free survival (RFS) were compared among the groups with different genotypes. RESULTS: (1) During the induction chemotherapy, XRCC3(C241T) polymorphic allele was significantly associated with the shorter survival of the AML patients with t (15; 17)/PML-RARalpha (OR = 8.750, P = 0.046). Among the non-M3 patients, the complete remission (CR) rate of those with double RAD51(G135C) and GSTT1 wild genotypes was 71.6%, significantly higher than that of those not with double RAD51(G135C) and GSTT1 wild genotypes (54.4%, P = 0.028). (2) The OS of the non-M3 AML patients with double RAD51(G135C) and GSTT1 wild genotypes was (39.1 +/- 7.1) months, significantly longer than those with double variant types [(22.4 +/- 3.2) months, P = 0.042]. The relapse-free survival (RFS) of the M4EO and M2 patients with double XRCC3(C241T) and GSTT1 wild type genotype were 48.3 months and 56.5 months, both significantly longer than those of the patients with double variant genotypes (28.8 months and 10.0 months respectively, both P < 0.05). The OS of the M2 patients with triple RAD51(G135C), GSTT1, and GSTM1 variant genotypes was (22.4 +/- 3.2) months, significantly shorter than those with triple RAD51(G135C), GSTT1, and GSTM1 wild genotypes [(39.1 +/- 7.1) months, P = 0.042]. The RFS of the M2 patients with triple RAD51(G135C), GSTT1, and GSTM1 variant genotypes was 10.0 months, significantly shorter than that of the patients with triple RAD51(G135C), GSTT1, and GSTM1 wild genotypes (64.2 months, P = 0.015). (3) The risk levels of neutropenia, nausea and vomiting, and alopecia of the patients with variant XRCC3(C241T) genotype were all significantly higher than those of the wild type genotype (all P < 0.05). The risk of hematuria of the patients with variant genotype was significantly higher than that of the patients with wild genotype (P = 0.017). CONCLUSION: The polymorphisms of XRCC3(C241T) and RAD51(G135C) genes are significantly related to response to therapy, adverse effects, and prognosis of AML. Detection of the XRCC3(C241T) and RAD51(G135C) genotypes may be useful in selecting individual chemotherapy regimens for patients with AML.

[Analysis of Early-Death and Factors Affecting Prognosis of Patients with Acute Promyelocytic Leukemia].

OBJECTIVE: To investigate the factors affecting the early-death, overall survival (OS) and relapse-free survival (RFS) of acute promyelocytic leukemia (APL) patients. METHODS: The clinical and laboratorial charachteristics of 176 APL patients in our center were analyzed retrospectively during January 2002 to Mar 2016. The risk factors of early death and factors affecting OS and RFS of patients were analyzed. RESULTS: Among total of 176 patients, early death occured in 10 patients. Multivariate analysis showed both age ≥60 years and fibrinogen<1.5 g/L (HR=6.4, 95%CI 1.4-28.2) (P=0.015), (HR=12.2, 95%CI 1.5-102.8) (P=0.021), respectively were the independent risk factors for the early death during the induction therapy. Among 154 patients with full follow-up data (median follow-up time was 101(2-262) months), the estimated 5-year OS and RFS rate were (98± 1)% and (77± 4)%, respectively. Cox regression analysis showed relapse during treatment as well as initial WBC count≥30× 109/L were independent prognostic indicators for OS. Accompanied psoriasis indicated higher relapse rate of APL(HR=4.8, 95%CI 1.8-12.5)(P=0.002), while the low-risk APL indicated lower relapse rate (HR=0.4, 95%CI 0.2-0.99)(P=0.048). CONCLUSION: Importance should be attached to the early-death events in elder and low-fibrinogen APL patients. As for patients with psoriasis or non low-risk group, emphasizing the intensified dynamic supervision during the treatment helps to detect the early-relapse events. For relapsed patients and patients with ≥30× 109/L WBC count, seeking more optimized therapy strategy seems allow this cohorts to get better prognosis.

[Chromosomal structural changes in patients with myelodysplastic syndrome].

OBJECTIVE: To study the clinical and laboratory features of myelodysplastic syndromes (MDS) with chromosomal structural changes. METHODS: Among 584 MDS cases with cytogenetic data, 50 patients with chromosomal structural changes, 34 males and 16 females, aged 50.5 (9 approximately 77), were reclassified according to the WHO criteria, and their clinical and laboratory features were analyzed retrospectively. RESULTS: The incidence of chromosomal structural changes in the MDS patients was 7.4%. i (17) (q10), t (1; 3) (p36; q21), der (1; 7) (q10; p10), and der (22) occurred frequently. The chromosomal structural changes in 13 cases were reported in the literatures for the first time. The patients with i (17) (q10) were characterized by moderate to severe anemia and a poor prognosis. Predominant dysgranulocytopoiesis and dysmegakaryocytopoiesis, including a nuclear shift to the left, pseudo-Pelger-Hüet anomaly, hypogranularity, and increased micromegakaryocytes. The patients with t (1; 3) (p36; q21) revealed macrocytic anemia, obvious dysmegakaryocytopoiesis, and dysgranulocytopoiesis, accompanied by defective differentiation and monocytosis. The bone marrow cells from the MDS patients with t (1; 3) (p36; q21) mainly or only expressed MEL1. The initial symptom of the patients with der (1; 7) (q10; p10) was infection. These patients showed macrocytic or normocytic anemia. Trilineage dysplasia was found in the bone marrow smears. The patients had short median survival. The patients with der (22) revealed anemia, and normal or elevated platelet counts. Hypogranularity and pseudo-Pelger-Hüet anomaly were present in all cases with der (22). The megakaryocytes were small and generally contained one or two nuclei. A translocation involvement of 22q11 was frequently found in the patients with der (22). CONCLUSION: MDS patients with i (17) (q10), t (1; 3) (p36; q21), and der (1; 7) (q10; p10) may be a new unique clinical-pathologic subsets. Whether the MDS patients with der (22) can be considered as a new unique subject remains to be confirmed by future studies.

[Role of Aberrant Splicing in Pathogenesis of Myelodysplastic Syndromes-Review].

The spectrum of genetic abnormalities in myelodysplastic syndromes(MDS) has been revealed by high-throughput sequencing. However, the functional role of these mutations in occurrence and development of MDS was not delineated. The mutations in splicing factors have been identified as the commonest gene mutations in MDS. Recently, it was reported that the mutated or dysregulated splicing factors, including SF3B1, SRSF2 and U2AF1, attribute to aberrant mRNA splicing, which leads to hematopoietic dysfunction and drives MDS. These findings will be of great help in searching for candidate therapeutic targets in mis-splicing pathway in MDS. In this review the role of aberrant splicing in pathogenesis of MDS is summarized briefly.

[The effects of glutathione S-transferase (GSTT1 and GSTM1) genes polymorphisms on treatment efficacy and prognosis of acute myeloid leukemia].

OBJECTIVE: To investigate the impact of GSTT1 and GSTM1 genotypes on response, drug side effects and prognosis of acute myeloid leukemia (AML). METHODS: GSTT1 and GSTM1 genotypes were analysed in 180 AML patients with PCR. Complete remission (CR) rate, drug side-effects, overall survival, relapse-free survival and relapse rate were compared in groups with or without GSTT1 and GSTM1 genes. RESULTS: (1) The CR rate (96.9%) in GSTT1 and GSTM1 genes double-present patients was significantly higher than that in patients of GSTT1 null or GSTM1 null (CR rate 78.0%) (P = 0.013). The risk of failure to achieve CR in patients with GSTT1 null/GSTM1 null is 8.736 times higher than that in patients with GSTT1 and GSTM1 genes double-present (odds ratio OR was 8.736, 95% CI was 1.146 - 66.574). The CR rate (88.4%) in GSTT1 present patients was also significantly higher than that in patients of GSTT1 null (CR rate 74.7%) (P = 0.021, OR = 2.572, 95% CI 1.136 - 5.826). (2) There was no significant relationship between GSTT1/GSTM1 genotypes and the lasting time of neutrophilic granulocyte (ANC) < 0.5 x 10(9)/L and PLT < 20 x 10(9)/L. The risk of ALT abnormality in patients with GSTM1 null is 2.593 times higher than that in patients with GSTM1 present (P = 0.016, 95% CI 1.176 - 5.717). (3) Overall survival and relapse-free survival of GSTT1 and GSTM1 double-present patients were significantly better than those in patients of GSTT1 null/GSTM1 null (mean overall survival was 68.4 months vs 38.5 months, P = 0.028, and mean relapse-free survival was 73.5 months vs 34.9 months, P = 0.014, respectively). Relapse-free survival in GSTT1 null patients was significantly shorter than that in patients with GSTT1 present (26.7 months vs 64.3 months, P = 0.038), but there was no significant difference of overall survival between the two groups. The relapse rate of double-present patients was significantly lower than that of GSTT1 null/GSTM1 null patients (13.3% vs 35.6%, P = 0.019). CONCLUSION: GSTT1 and GSTM1 genotypes were apparently related with response, drug side effects and prognosis of patients with AML. GSTT1 and GSTM1 genotype might be useful in selecting appropriate chemotherapy regimens for patients with AML.

[An analysis of cytogenetic characteristics and prognosis of 189 t (8; 21) acute myeloid leukemia patients].

OBJECTIVE: To investigate the cytogenetic and prognostic significance of acute myeloid leukemia (AML) with t (8; 21). METHODS: 189 patients with t (8; 21) AML were categorized according to their additional karyotypic aberration and their clinical outcomes analysed. RESULTS: Among them, 63 patients (33.3%) were t (8; 21) without other additional aberrations, 126 cases (66.7%) were t (8; 21) with other additional aberrations. -Y was found in 46.7% (63/135) of the male and -X was found in 25.9% (14/54) of female patients. In additional aberrations, loss of the sex chromosome were found in 77 cases (61.1%), Del (9q) was found in 16 cases (12.7%), +4 was found in 5 cases (4.0%); 7q- was found in 6 cases (4.8%); Tetraploidy (4N) was found in 2 cases (1.6%); Variant translocation was found in 7 cases (5.6%). The 189 patients had a high remission rate (87.0%) and a relatively long median survival (21.6 months). +4 and 4N were an unfavorable prognostic factors. Fluorescence in situ hybridization technique is a more sensitive and accurate method to detect t (8; 21), especially in variant translocation, complex variant translocation and masked translocation. CONCLUSION: t (8; 21) AML is also frequently associated with additional chromosome aberrations, these aberration had influence on prognosis.

An effective and simple expansion system for megakaryocyte progenitor cells using a combination of heparin with thrombopoietin and interleukin-11.

Transfusion of ex vivo expanded megakaryocyte (MK) progenitor cells has been suggested to shorten the time of platelet recovery in the thrombocytopenia induced by radiotherapy or chemotherapy. Here, we report an effective and simple expansion system of MK progenitor cells from cord blood (CB) CD34(+) cells using a combination of thrombopoietin (TPO), interleukin (IL)-11, and heparin. When the CB CD34(+) cells were cultured in a liquid expansion system in the presence of TPO + recombination human (rh)IL-11 + heparin for 7 days, the number of CB CD34(+)/CD41a(+) cells was significantly increased compared to control groups (p < 0.05). When the suspension cells collected from 7-day liquid culture were replated in semisolid cultures, increased large MK colonies were observed in the culture with combination of TPO + IL-11 + heparin compared to those of control groups. In vivo, transfusion of CD34(+) cells expanded with TPO + IL-11 + heparin into irradiated nonobese diabetic/severe combined immunodeficient mice significantly accelerated platelet recovery. These data indicate that heparin as effective cofactor for TPO and IL-11 promotes expansion of MK progenitor cells from CB CD34(+) cells. This expansion system is simple and effective and could be used for the treatment of thrombocytopenia after radiotherapy or chemotherapy.

Long-term outcomes of imatinib in patients with FIP1L1/ PDGFRA associated chronic eosinophilic leukemia: experience of a single center in China.

BACKGROUND: The FIP1L1/PDGFRA (F/P) fusion gene is the most common clonal genetic abnormality of chronic eosinophilic leukemia (CEL). Tyrosine kinase inhibitors (TKI), such as imatinib, have been demonstrated to be effective therapies for F/P mutated disease. The aim of this study was to analyze the treatment response and long term prognosis in patients with F/P mutated CEL. METHODS: The clinical features and treatment responses of 33 consecutive patients with F/P mutated CEL between August 2006 and October 2014 were analyzed. The 33 cases received imatinib therapy at an initial dose of 100 mg/day (30 patients) or 200 mg/day (3 patients); the maintenance dose depended on the response condition and patient willingness. Through the follow up, the molecular responses were regularly monitored. RESULTS: With a median follow up of 64 months, 94% of the 33 patients with F/P mutated CEL achieved a complete hematologic remission (CHR), and 97% achieved a complete molecular remission (CMR) after a median of 3 (1.5-12) months. Twenty-four cases received maintenance therapy, with a median CMR duration of 43 (5-88) months. Imatinib therapy was discontinued in 8 cases, including 4 cases who experienced relapse, and 4 patients who maintained CHR or CMR after discontinuing therapy with a median time of 47 (2-74) months. One case exhibited primary resistance with a PDGFRA T674I mutation. CONCLUSIONS: F/P mutated CEL has an excellent long-term prognosis following imatinib therapy. A 100 mg daily dose of imatinib is sufficient to induce remission, and a single 100 mg weekly dose maintains a durable remission. A subgroup of patients may maintain a durable remission after discontinuing therapy with a CMR.

[Relationship between GSTT1, GSTM1 and NQO1 gene polymorphism and acute myeloid leukemia and recurrent chromosome translocations].

OBJECTIVE: To investigate the impact of GSTM1, GSTT1 and NQO1 genotypes on susceptibility to acute myeloid leukemia (AML) and recurrent chromosome translocations of AML. METHODS: GSTT1, GSTM1 and NQO1 genotypes were detected in 228 adult patients with de novo AML and 241 controls by PCR or PCR-RFLP. RESULTS: The frequency of GSTM1 null genotype in the AML patients was 62.3%, significantly higher than that in the normal controls (52.7%, P = 0.036), however, no significant difference was found in the incidence of GSTT1 null genotype. The frequencies of NQO1(C609T) C/T and T/T genotypes were 53.1% and 25.0% respectively among the total AML patients (53.1% and 25.0% respectively), 64.3% and 25.0% respectively among the AML patients with t (8; 21) (q22; q22)/AML-ETO fusion gene, and 57.1% and 26.0% respectively among the AML patient with t (15; 17) (q22; q11)/PML-RARalpha fusion gene, all significantly higher than those in the controls (49.4% and 13.7% respectively). The relative risk of t (8; 21) (q22; q22)/AML-ETO (+) AML was 4.487 (95% CI: 1.282-15.705) for the subjects with NQO1(C609T) C/T genotype, and was 6.293 (95% CI: 1.536-25.782) for the subjects with NQO1(C609T) T/T genotype. The relative risk of t (15; 17) (q22; q11)/PML-RARalpha (+) AML was 2.531 (95% CI: 1.286-4.981) for the subjects with NQO1(C609T) C/T genotype, and was 4.149 (95% CI: 1.856-9.275) for the subjects with NQO1(C609T) T/T genotype. CONCLUSION: Determination of the NQO1(C609T) genotypes may be used as a stratification marker to predict high-risk individuals for AML, especially for AML with t (8; 21) (q22; q22)/AML-ETO fusion gene and t (15; 17) (q22; q11)/PML-RARalpha fusion gene.

[In vitro expansion of cord blood megakaryocyte progenitor].

OBJECTIVE: To expand cord blood megakaryocyte progenitor cells in vitro. METHODS: Cord blood CD34+ cells were selected by magnetic cell sorting (MACS), and thrombopoietin (TPO), interleukin-11 (IL-11), and heparin were used in the expansion system of megakaryocyte progenitor. The expansion efficiency was measured by fluorescence-activated cell sorting (FACS) using the megakaryocytic specific monoclonal antibodies (CD34+, CD41a+, CD61+, CD34+CD41a+, CD41a+CD61+) and colony-forming units-megakaryocyte (CFU-MK) analysis. The expanded megakaryocyte progenitor were determined by histochemistry staining using CD41a and the observation of the ultrastructure of megakaryocyte (MK) by electron microscopy. The megakaryocyte function were examined by the platelet activation in vitro and nonobese diabetic/severe combined immunodifficiency (NOD/SCID) mice transplantation in vivo. RESULTS: CD34+CD41a+ cells was expanded (4.0 +/- 1.7) folds on day 7 in TPO (50 ng/ml) group and (10.5 +/- 4.8) fold in TPO combined with IL-11 group; after heparin was joined in on day 0, a more significantly elevated expansion was found in the heparin, TPO, and IL-11 group [(29.9 +/- 6.4) folds than the above two groups; P < 0.05]. Meanwhile, the large CFU-MK colony (> 50 cells/colony) was (106.8 +/- 26.9) folds on day 7 (P < 0.05). The megakaryocyte expanding with TPO, IL-11 and heparin for 7 days in vitro transplanted the NOD/SCID mice fasten the recovery of platelet and white blood cell account and improved the survival. Megakaryocyte under culture displayed certain development of territories membrane. Platelet activation test comfirmed that the expanding megakaryocyte progenitor had the normal function. CONCLUSION: TPO, IL-11, and heparin combination system for ex vivo expansion is an effective expansion system of megakaryocyte progenitor.

Autologous transplantation of peripheral blood stem cells as an effective therapeutic approach for severe arteriosclerosis obliterans of lower extremities.

Treatment of severe arteriosclerosis obliterans of lower extremities (ASOLE) remains a clinical challenge. To develop a more effective approach, we evaluated the clinical efficacy of autologous transplantation of mobilized peripheral blood stem cells (PBSCs) in 5 patients with ASOLE. The patients received recombinant human granulocyte colony-stimulating factor (rhG-CSF, 600 micro g/day) for 5 consecutive days. On day 5, PBSCs were collected, sorted from blood circulation of patients, and then intramuscularly injected into their ischemic lower limbs. A significant improvement of clinical manifestations including severe pain, skin temperature and ulcer, was observed, without obvious adverse effect. The patient's limb was successfully saved. Satisfactory remission was obtained 3 months after transplantation as shown by significant improvement in ankle-brachial pressure index (ABI), blood flow in personal vascular laboratory (PVL), laser Doppler blood perfusion, and the angio-graphic scores. Our data suggest for the first time that autologous transplantation of mobilized PBSCs provides a practical, safe, and effective method of treatment for lower limb ischemia.

[The relevance between quantitative and type of chromosomal abnormality and leukemia transformation in myelodysplastic syndrome].

OBJECTIVE: To investigate leukemia transformation rate in myelodysplastic syndrome (MDS) and the relationship with quantitative and type of chromosomal abnormality. METHODS: This study retrospectively analyzed and rediagnosed 138 MDS patients with complete data, investigated the rate and time of leukemia transformation, and analyzed characteristics of chromosome karyotype of de novo patients. RESULTS: 29 (21.01%) of 138 patients transformed into leukemia, the rate and the median time of leukemia transformation were 21.01% and 8 (3-24) months, respectively, among which, the rate of leukemia transformation in normal karyotype, abnormal karyotype analysis of ≤5 mitotic cells, and >5 mitotic cells in split phase groups were 6.2%, 23.8% and 38.5%, respectively, and median time of which were 17(13-22), 13(5-23), and 7(3-10) months, respectively. Increased trend of leukemia conversion rate along with increased quantity of chromosomal abnormality was observed (χ²=14.185, P＜0.01). Leukemia transformation time negatively correlated with quantity grade of abnormal karyotype (r=-0.631, P<0.01), The leukemia transformation rates in monosomy 7/del 7q, trisomy 8, trisomy 11, complex karyotype and normal karyotype groups were 65.0%, 50.0%, 30.8% and 28.6%, being significantly different (χ²=21.555, P<0.01). Leukemia transformation rate of complex karyotype and monosomy 7/del 7 q was slightly higher than of trisomy 8 and trisomy 11, but both of them were significantly higher than of normal karyotype (χ²=8.054, P=0.005). There were no leukemia transformation cases in del 5q, del 20q, monosomy Y, and trisomy 21 group. CONCLUSION: With or without abnormal chromosome karyotype, quantity and types of abnormal karyotype had important clinical value to predict leukemia transformation in patients with MDS.

[The effect of transmission electron microscopy on diagnosis of acute myeloid leukemia].

OBJECTIVE: To analyze coincidence rate of acute myeloid leukemia (AML) sub-typing between transmission electron microscopy (TEM) and clinical discharge diagnosis. METHODS: Reviewing sub-typing results of TEM, light microscopy, flow cytometric analyzing, molecular biological detection and karyotype in 793 AML cases, comparing their coincidence rates with discharge diagnosis to reveal advantages of AML sub-typing by TEM. RESULTS: General coincidence rates of TEM, light microscopy, flow cytometric analyzing, molecular biological detection and karyotype on AML sub-typing were 63%, 59%, 52%, 47%, 26% and 23% respectively, and clinical coincidence rates of TEM on M1, M2a, M4 and M5, M6, M7, t (8; 21) and t (15; 17) were 39%, 34%, 17%, 74%, 50%, 73%, 87% and 89% respectively. CONCLUSION: TEM has a higher coincidence rate in general AML sub-typing, especially strong screenings on t (15; 17), t (8; 21), M7, M5 and M6, but lower coincidence rates on M1, M2a and M4 sub-typing than other methods.

Distinct clinical and experimental characteristics in the patients younger than 60 years old with myelodysplastic syndromes.

Myelodysplastic syndromes (MDS) mainly occur in elderly individuals in Western countries. However, MDS is commonly found in young individuals (<60 years) in Asia. The reason for the high incidence in younger individuals is still unclear, and the differences in disease features between young and elderly patients with MDS have been not well recognized. To explore these issues, in this study, we analyzed the clinical and experimental characteristics of MDS in the patients younger and older than 60 years old and characterized the potential age-associated differences. The results showed that over half of the patients with MDS (61.9%) were younger than 60 years old upon the first diagnosis. The younger patients were more likely to be female, who have lower risk and less advanced MDS. The occurrence of trisomy 8 and bone marrow failure were more frequent in the younger patients than the older ones. The marrow CD34+ cells in the younger patients showed lower proliferation and higher apoptosis in comparison with that in the older ones. Obvious amplification of T cells and low CFU formation could be found in the younger patients. CFU formation was significantly increased in the younger patients after the removal of activated T cells. In addition, the younger patients had a lower frequency of p15(INK4B) methylation, longer survival expectancy and less AML transformation. In summary, the younger patients with MDS in China may show more benign disease features than the older ones. Enhanced immunological response may be involved in the pathogenesis of MDS in the patients younger than 60 years.

[Relationship between polymorphisms of tumor necrosis factor alpha gene and primary myelodysplastic syndromes].

OBJECTIVE: To investigate the association of single nucleus polymorphisms(SNP)of tumor necrosis factor alpha (TNF-α) gene (-308 G>A and -238 G>A genotypes) with susceptibility to primary myelodysplastic syndromes (MDS). METHODS: Two SNPs (TNF-α-308 G>A,TNF-α-238 G>A) of TNF-α gene were detected by Taqman probes in 341 MDS patients and 365 unrelated-healthy controls. RESULTS: Compared to healthy controls, the frequency of TNF-α-308 AA+AG genotype and A allele increased (18% vs 10%, P=0.015, 9% vs 5%, P=0.021, respectively) in refractory cytopenia with multilineage dysplasia (RCMD) patients. There was no correlation of TNF-α-308 G>A genotype and allele frequency between MDS and controls. No difference in the genotype and allele frequency of TNF-α-238 G>A were found between controls and MDS or the subtypes of MDS (P>0.05). We did not find any linkage between plasma level of TNF-α and TNF-α-308 G>A or TNF-α-238 G>A genotype. Statistic differences were observed between platelet count[58(1-611)×109/L vs 90(7-352)×109/L]and bone marrow blasts in MDS patients carrying TNF-α-308 G>A GG and AA+AG genotype (P=0.024, 0.019, respectively). CONCLUSION: TNF-α-308 G>A polymorphism was correlated with susceptibility to MDS-RCMD.