Characteristics and Clinical Significance of Gene Mutation in Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

OBJECTIVE@#To investigate the characteristics of gene mutations in patients with myelodysplastic syndromes (MDS) and its prognostic significance.@*METHODS@#High-throughput sequencing was used to detect 34 blood tumor-related genes in 210 patients with MDS, and the relationship with the revised International Prognostic Scoring System (IPSS-R) and the impact on prognosis of the patients were analyzed.@*RESULTS@#Among the 210 MDS patients, 142 cases (67.6%) showed mutations, and the first six genes with the highest mutation detection rate were ASXL1(20.5%), TET2(17.1%), U2AF1(14.3%), DNMT3A (11.9%), TP53(10.5%) and RUNX1(10.0%). The gene mutation rate of the patients in IPSS-R relatively high-risk group was higher than those in relatively low-risk group (P=0.001). Both TP53 and BCOR genes showed higher mutation rates in the higher risk group than in the lower risk group (P<0.05). Survival time of the patients in TP53 mutant group was lower than those in non-mutant group (P<0.001), survival time of patients in SF3B1 mutant group was higher than those in non-mutant group (P=0.018). According to the number of gene mutations, the patients could be divided into groups with 0-1, 2 and ≥3 gene mutations, and the median OS of the three groups were not reached, 43 and 27 months, respectively (P=0.004). The Multivariate analysis showed that the increasing number of gene mutations and TP53 mutation was the independent risk factors affecting prognosis of the patients, while SF3B1 mutation was the independent protective factor for the prognosis of the patients.@*CONCLUSION@#The gene mutation rate was higher in MDS patients. And the increasing numbers of gene mutation, TP53 and SF3B1 were the influence factors of prognosis in the patients.

Effect of Sonic Hedgehog Signal Pathway Inhibitor Jervine on Myelodysplastic Syndromes MUTZ-1 Cells / 中国实验血液学杂志

OBJECTIVE@#To study the effect of SMO inhibitor (Jervine) on proliferation, apoptosis and cell cycle of MDS cell line MUTZ-1, and its mechanism.@*METHODS@#The effect of different concentrations Jervine on proliferation of MUTZ-1 cells was detected by CCK-8 method. Apoptosis and cell cycle of MUTZ-1 cells were detected by flow cytometry. Western blot was used to detect the changes of Shh signaling pathway effecting proteins BCL2 and CyclinD1. The expression levels of Smo and Gli1 gene were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR).@*RESULTS@#Jervine inhibited MUTZ-1 cell proliferation in a concentration dependent manner (24 h, r=-0.977), the apoptosis rate of MUTZ-1 cells increased with the enhancement of concentration of Jervine in MUTZ-1 cells (P＜0.001), the cell proportion of G phase increased and the cell number of S phase decreased with enhancement of concentration (P＜0.001). The result of RT-qPCR and Western blot showed that the expression of Smo, Gli1 mRNA and BCL2, CyclinD1 proteins decreased (P＜0.05).@*CONCLUSION@#SMO inhibitor can effectively inhibit the growth of MDS cell line MUTZ-1 improve the cell apoptosis and induce cell cycle arrest. Its action mechanism may be related with dowm-regulating the expression of BCL2 and CyclinD1.

Expression Level and Clinical Significance of Gli1 in Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

OBJECTIVE@#To study the expression level and clinical significance of Gli1 gene in patients with myelodysplastic syndrome(MDS).@*METHODS@#The positive rate of bone marrow CD34 cells was detected by flow cytometry in 53 patients with MDS.Magnetic beads were used to separate CD34 cells. The expression of Gli1 on CD34 cells was detected by RT-qPCR, 25 patients with iron deficiency anemia were selected as controls. The relationship of Gli1 expression with clinical characteristics were analyzed.@*RESULTS@#The expression of Gli1 in patients with MDS (0.73±1.26) was significantly higher than that in the control group (0.07±0.46) (P＜0.05). The expression of Gli1 significantly correlated with platelet count, chromosome grouping and IPSS risk stratification (P＜0.05). The median overall survival time of patients in high and low expression groups were 7 and 20 months respectively (P＜0.05). Multivariate analysis showed that Gli1 and chromosome grouping were 2 independent poor prognostic factors (P＜0.05).@*CONCLUSION@#The expression of Gli1 is high in MDS. Abnormal expression of Gli1 positively correlates with clinical characteristics and prognosis of patients.Gli1 may be involved in the occurrence and development of MDS.

Prognostic Value of Multigene Methylation in Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the methylation status of p15, DAPK, SOCS1 and FHIT genes in patients with myelodysplastic syndrome(MDS) and to explore the prognostic significance of gene methylation.</p><p><b>METHODS</b>Methylation-specific PCR (MSP) was used to detect the methylation of the above-mentioned 4 genes in 67 patients with MDS and 18 patients with iron-deficient anemia as controls. The gene methylation status of MDS patients and its effect on prognosis were analyzed.</p><p><b>RESULTS</b>The methylation rates of p15, DAPK, SOCS1 and FHIT in 67 MDS patients were 37.3%, 35.8%, 47.8% and 52.2%, respectively, which were significantly higher than those in the control group (P<0.05). The methylation status of p15, SOCS1 was increased along with the increase of International Prognostic Scoring System(IPSS) scores (P<0.05), and ≥2 genes was more frequent in relatively high risk groups (P<0.05). The median overall survival time of patients with and without methylation were 15 and 21 months, respectively (P<0.05). Patients showing methylation of SOCS1 had a significantly shorter survival time in relatively low risk groups(P<0.05), meanwhile SOCS1, p15 and methylations of ≥2 genes had significantly shorter survival time in relatively high risk groups(P<0.05). In multivariate analysis, SOCS1 and p15 were negative prognostic factors.</p><p><b>CONCLUSION</b>p15, DAPK, SOCS1 and FHIT are higher hypermethylated genes in MDS. The methylations of SOCS1 and p15 are independent prognostic factor for overall survival in MDS.</p>

Clinical Value of p15, DAPK, SOCS1 and FHIT Genes Combined Detection in the Early Diagnosis and Prognosis Evaluation of Myelodysplastic Syndrome / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the value of p15, DAPK, SOCS1 and FHIT genes combined detection in the early diagnosis and prognosis evaluation of patients with myelodysplastic syndrome(MDS).</p><p><b>METHODS</b>The methylation-specific PCR (MSP) was used to detect the methylation of the above-mentioned 4 genes in 67 patients with MDS. The value of 4 gene combined detection in the early diagnosis and prognosis evaluation of patients with MDS was compared and anazlyzed.</p><p><b>RESULTS</b>The methylation rates of p15, DAPK, SOCS1 and FHIT genes in 67 patients with MDS were 37.3%, 35.8%, 47.8% and 52.2%, respectively, which were significantly higher than those in control group (P<0.05). The accordance rates of p15, DAPK, SOCS1 and FHIT single detection for diagnosis of MDS were 37.3%,35.8%,47.8% and 52.2%, respectively, meanswhile the accordance rate of above-mentioned 4 gene combined detection for diagnosis of MDS was 82.1%, which was significantly higher than that of single gene detection(P<0.001). The methylation of ≥2 genes in relatively high risk group was significantly higher than that in relatively low risk group (P<0.05). The median survival time of MDS patients was 18(13.3, 22.7) months; the median survival time in relatively low risk group was significantly longer than that in relatively high risk group [27(20.3,33.7) months vs 9(5.9,12.1) months] (P<0.05). The survival time of MDS patients with different risks displayed the trend of shorting feature along with increasing of methylated genes (P<0.05).</p><p><b>CONCLUSION</b>The combined detection of above menthioned 4 genes can improve the accuracy of early diagnosis and prognosis evaluation for MDS patients.</p>

Analysis of Therapy and Efficacy after Remission of Acute Myeloid Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the therapy and efficacy after remission of patients with acute myeloid leukemia (AML).</p><p><b>METHODS</b>The clinical data of 110 patients diagnosed as AML treated from 2008 to 2013 were analyzed retrospectively. According to different consolidation therapy regimens, the patients were divided into 4 groups:1 ID-Ara-C group, 2 ID-Ara-C group, 3-4 ID-Ara-C group, allogeneic hematopoietic stem cell transplantation (allo-HSCT) group. The disease-free survival (DFS) and overall survival (OS) were analyzed retrospectovely.</p><p><b>RESULTS</b>110 patients were completely remittend by 1-2 couses. The median follow-up time was 26.8 months, 35 cases relapsed and 58 cases died, the median DFS and OS were 20.5 and 26.8 months. The 3-year DFS of 1, 2, 3-4 ID-Ara-C and allo-HSCT groups were 0%, 36.1%, 37.5% and 67.9% respectively. The 5-year DFS of 1, 2, 3-4 ID-Ara-C and allo-HSCT groups were 0%, 30.1%, 37.5% and 63.0%. The 3-year OS rates of 1, 2 ID-Ara-C groups, 3-4 ID-Ara-C group and allo-HSCT group were 24.0%, 36.0%, 58.3% and 67.8%. The 5-year OS of 1, 2 ID-Ara-C grous, 3-4 ID-Ara-C and allo-HSCT group were 24.0%, 36.0%, 58.3% and 67.8% respectively. The 5-year OS rates of 1 ID-Ara-C group, 2 ID-Ara-C group, 3-4 ID-Ara-C group were 0%,30.0%,35.0% and 62.9% respectively. The multifactor analysis indicated that the courses of ID-Ara-C and allo-HSCT were independent risk factors for DFS and OS.</p><p><b>CONCLUSION</b>≥2 ID-Ara-C regimen may be used as one regimen of consolidation therapy for patients with AML after remission.</p>

Serum Ferritin Level - Risk Factors of Myelodysplastic Syndrome Prognosis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate whether serum ferritin (SF) level may be used as a indicator for predicting mortality of patients with myelodysplastic syndrome (MDS).</p><p><b>METHODS</b>A total of 151 patients with MDS were followed up in our study, their blood routine indicators, bone marrow blasts and SF level were detected. All of the patients were divided into the dead group and survival group.</p><p><b>RESULTS</b>The average survival time of all patients was 30.0 ± 10.86 months. There were statistical differences in age, IPSS score, chromosome grouping and SF level between 2 groups (P < 0.05). COX model analysis showed that age, MDS type, IPSS score, chromosome grouping and SF level all were related with mortality of patients with MDS, which were risk factors of death for patients with MDS (P < 0.05). The receiver-operating characteristic curve (ROC) area for SF was 0.826 with a Cut off value of 622.95, the sensitivity and specificity was 77.5% and 75% respectively. Log-rank test showed that the mortalities of patients with different levels of SF were statistically and very significantly different (P < 0.01).</p><p><b>CONCLUSION</b>The age, IPSS score, chromosome grouping and SF level closely correct with mortality of the patients with MDS, the SF level may be considered as a predictor of death for MDS patients.</p>

Comparison of chromosome karyotype between myelodysplastic syndrome and acute leukemia patients confirmed at the same period / 中国实验血液学杂志

This study was purposed to compare and analyze the relationship between the abnormality of chromosome karyotypes and diagnosis, prognosis of MDS and AML patients, as well as to explore the characteristics of chromosome prognostic stratification in MDS and AML patients of different ages. The cytogenetic karyotype analysis was performed in 134 cases of MDS and 123 cases of AML by using bone marrow short-term culture and R-banding technique. The results indicated that the detected rates of chromosome abnormal karyotypes in MDS and AML patients were 41% and 61% respectively. The abnormal karyotype analysis of MDS and AML group showed that the abnormal karyotype in MDS group displayed number abnormality as the dominate (mainly the +8), while the abnormal karyotype in AML group displayed structure abnormality as the dominant [mainly, t(15;17) and t(8;21)]. The detected abnormal karyotype are mainly for the +8 which has ambiguous correlation with FAB subtype; the detection rates of complex karyotype abnormalities, favourable prognosis karyotype as well as poor prognosis karyotype in the MDS group obviously higher than that of AML group. Among patients with MDS transformed into AML, 12 cases had chromosome abnormal karyotype. There were 3 cases of chromosome abnormal karyotype in AML group which were transformed by MDS. The analysis of age stratification between two groups showed that the detected rate of abnormal karyotype was enhanced with the increase of age in MDS group, and detected rate in ≥ 60 years old group was obviously higher than that in patients with ≤ 30 age group.The detected rate of complex karyotype abnormalities in three age groups of MDS did not show statistical difference; the detected rate of abnormal karyotype in AML group decreased with the increase of age, the detected rate in ≤ 30 years old group was obviously higher than that in ≥ 60 age group,while the detection rate of complex karyotype abnormalities showed that the detected rate in patients ≥ 60 years old group was obviously higher than that in patients with ≤ 30 years old group; Analysis of karyotype prognosis revealed that the detected rate of poor prognosis karyotype increased along with the age growth both in MDS and AML groups, and detected rate in ≥ 60 years old group was obviously higher than that in ≤ 30 years old group; while analysis of favourable prognosis karyotype in MDS and AML group showed that the detected rate in ≤ 30 years old group was obviously higher than that in ≥ 60 years old group. It is concluded that the patients with MDS and AML have higher chromosomal abnormalities,which have important reference value for the diagnosis, treatment and prognosis, meanwhile, the analysis of chromosome karyotype provides an important basis for prognostic stratification.

Comparison on main growth characters of test-tube plants and seeding plants in Bupleurum chinense / 中草药

Objective: Analyzing hereditary parameters and comparing the test-tube plants population of Bupleurum chinense and seeding plants in different growing periods to provide the basis for building and screening out the right way to foster superior seeds of B. chinense. Methods: The test-tube plants were transplanted into the croplands and the same type of seeds were sowed at the same time. Until the next year, after the reviving of the plants, the plants were marked and the timing measure on the data of number of tiller, number of branches, number of leaves, length of leaves, width of leaves, number of jointings, length of jointing, sum of stem thickness, plant height, and number of flowers was carried out. Then, the roots were dug up and the medical characters of the roots, for instance, length, degree of thickness, and weight of the roots were measured. Following that, the statistic analysis on the data was conducted by using the software of SPSS 20.0. Results: The relative standard deviation values of test-tube plants population of B. chinense in diverse genetic characters were all relatively small. Moreover, plants were tidy, individual difference was tiny, and genetic characters were stable. In addition, the relevance of the characters of test-tube plants in various growing periods had some differences with the seeding plants. At last, compared with the seeding plants, the test-tube plants populations of B. chinense had more obvious superiority on the yield of roots and output of seeds. Additionally, the medicinal characters and quality were quite even. Conclusion: Compared with the seeding plants, the test-tube plants population of B. chinense has the superior botanical and pharmacognosy characters. Moreover, plant tissue culture technique is an effective way to breed and enlarge the superior seeds of B. chinense.

Comparison of bone marrow and blood cell morphology between refractory anemia and other anemia disease / 中国实验血液学杂志

This study was purposed to investigate the cell morphological features of bone marrow and peripheral blood in patients with myelodysplastic syndrome, mainly with refractory anemia, and to compare them with other anemia diseases including chronic aplastic anemia, hemolytic anemia and megaloblastic anemia. The bone marrow and peripheral blood were taken from patients for preparing the smears with Wright staining. 500 karyocytes in bone marrow and 100 karyocytes in peripheral blood were detected, and the features of morbid cells of erythrocyte, granulocyte and megakaryocytic series were observed. The results showed that differences between refractory anemia, chronic aplastic anemias and hemolytic anemia as well as megaloblastic anemia were statistically significant (P < 0.05) in the granules scarce and absence in the intracytoplasm of segmented neutrocyte in peripheral blood, Pelger dyskaryosis, the numbers and detected rate of immature granulocytes, monocyte detected rate, the granules scarce in all stage of granulocytic series in bone marrow, odd number and prolification of nucleolus in erythrocytic series, little macronucleus and single circle nucleus macronucleus. It is concluded that cell morphology is the foundation of diagnosing the MDS, the abnormality morphology both in peripheral blood and bone marrow play the consequence role in the diagnosis of MDS.

Non-T-cell depleted HLA haploidentical peripheral blood stem cell transplantation for hematological malignancies: report of 36 cases / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the clinical outcome of human leukocyte antigen (HLA) haploidentical peripheral blood stem cell transplantation (PBSCT) from related donors for hematological malignancies.</p><p><b>METHODS</b>Thirty-six patients with hematological malignancies, with a median age of 25 (11-48) years, were transplanted with PBSC from an HLA-haploidentical family donors: 7 were 1 locus mismatched and 29 were 2-3 loci mismatched. The recipients received myeloablative conditioning regimen, in combination with different immunosuppressants according to the degree of HLA disparity followed by non-T-cell depleted PBSCT. The median number of CD34+ cells were 11 (4.16-21.00) x 10(6)/kg.</p><p><b>RESULTS</b>All patients achieved sustained, full donor-type engraftment. Fifteen patients (41.7%) developed grade I-II aGVHD. Among 29 patients followed up more than 18 months, 17 (58.6%) developed cGVHD. There was no statistical difference in decrease and recovery of T, B and NK cell subsets after transplantation between HLA haploidentical group and HLA identical PBSCT group. The median follow-up duration was 15 (4 -69) months. Five patients (13.9% ) relapsed. The 2-year probability of leukemia-free survival (LFS) was 82.2%.</p><p><b>CONCLUSION</b>Non-T-cell depleted HLA-haploidentical PBSCT is safe and feasible for patients with hematological malignancies after myeloablative conditioning regimen combined with intensive immunosuppressants.</p>

Retinoic acid in treating acute promyelocytic leukemia with hyperleukocytosis and its therapeutic strategy / 中国实验血液学杂志

In order to investigate the occurrence of hyperleukocytosis in treating acute promyelocytic leukemia (APL) patients with all trans retinoic acid (ATRA) and to explore the influence of the level of leucocyte on curative effect of ATRA, the APL patients were divided into three different groups according to the count of leucocyte in peripheral blood. Patients with WBC count less than 30x10(9)/L were administered with ATRA alone (the first group), patients with WBC count more than 30x10(9)/L were administered with ATRA alone (the second group) and patients with WBC count more than 30x10(9)/L were treated with ATRA+cytotoxic drugs (the third group). The results showed that hyperleukocytosis were found in 23 out of 39 patients (58.97%). Total remission rates in the second group and in the third group were 91.3%. The remission rates in the first, second and third groups were 100%, 87.5% and 90.9%, respectively. It is concluded that the ATRA in combination with cytotoxic drugs can efficiently control the occurrence of hyperleukocytosis during ATRA-treating APL and reduce the early mortality.

Determination of content of saikoside in Bupleurum Chinense from different sources / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the contents of saikosaponin a, d and saikosaponins in Radix Bupleuri from different and assess their quality.</p><p><b>METHOD</b>Saikosaponin a and d were used as the chemical reference substance to establish RP-HPLC and visible-spectrophotometry methods for determination of contents of saikosaponin a, d and saikosaponins.</p><p><b>RESULT</b>The quality of B. chinense was better than that of B. scozonerifolium and B smishii; The contents of a, saikosaponin d and total saikosaponin in B. chinense were 0.100%-0.330%, 0.200%-0.400% and 1.00%-2.00% , respectively.</p><p><b>CONCLUSION</b>It is to suggest that the minimal contents requirements of saikosaponin a, d and saikosaponins in B. chinense should be set at 0. 100% , 0.200% and 1.00% , respectively; the best suitable planting areas are Taigu in the middle of Shanxi province, Ruicheng, Pinglu and Wanrong in the south of Shanxi province, and Longxi, Lixian in Gansu province.</p>

CD34+ antigen expression relating to prognosis in acute myeloid leukemia / 中国实验血液学杂志

To explore CD34(+) antigen expression in new diagnosed acute myeloid leukemia (AML) and analyze the prognosis for CD34(+) AML patients, the expression of antigen CD34 in 238 AML patients was detected by indirect immunofluorescence assay. The results showed that CD34 in 92 out of the 238 patients (38.7%) were positive, there was relationship between the CD34(+) expression and FAB subtypes (M(0), M(1)), and no CD34(+) expression was observed in M(3) subtypes. The complete remission rate of CD34(+) AML patients was 32%, which was lower than that of CD34(-) AML (61%). The lymphoid-associated antigen (CD7) was significantly increased in CD34(+) AML patients, compared with CD34(-) patients (P < 0.05). It is concluded that CD34(+) AML patients show poor prognosis and lower CR rate. The detection of CD34 expression is of some value in predicting prognosis in AML.

Effects of genistein and 17?-estradiol on microstructure of cancellous bone in ovariectomized rats / 中华内分泌代谢杂志

Objective To observe the effects of genistein and 17?-estradiol on microstructure of cancellous bone in ovariectomized (OVX) rats.Methods Ninty 7-month-old SD rats were randomly divided into baseline group,ovariectomized (OVX),sham-operated (SHAM),17?-estradiol treated (10?g?kg~(-1).day~(-1),EST) and genistein treated (5 mg?kg~(-1)?day~(-1),GEN) groups,and were killed at the beginning of the experiment,the 3rd and 15th week after operation.MicroCT scanning was performed on the left tibia in vitro.The regions involving 0.5 mm slice thickness and 1.6 mm distal to the tibial growth plate were selected as the regions of interest.Results At the 3rd week after operation,the tissue bone mineral density (tBMD) and trabecular thickness (sTh.Th) in group GEN were significantly higher than those in OVX and EST groups (all P