Objective To explore the clinical significance of expression of the CD20 in 96 adults B-lineage acute lyrnphoblastic leukemia (B-ALL). Methods The CD20 expression of 96 acute lymphoblastic leukemia patients were determined by flow cytometry. The characteristics ,examination results and outcome were analyzed retrospectively. Results Out of the 96 patients, there were 29 (30.20 %) patients with CD20positive and 67 (69.79 %) patients with CD20 negative. The distribution of age, infiltration of liver, spleen, and lymphnodes, the expression of myeloid lineage marker, the incidence of Ph chromosome and bcr-abl fusion gene and the complete remission rate within 4 weeks between CD20 positive and negative groups showed no significant differences (P ＞ 0.05). The relapse rate and 3 year over survival rate of adults B-ALL in CD20 positive and negative groups were 54.55 % and 14.80 %, 29.63 % and 37.30 % respectively with a significant differences (x2 = 0.42, 5.31, P＜ 0.05). Conclusion The expression of CD20 in adult B-ALL appears to be not associated with clinical features and CD20 expression in adult B-ALL cells appears to be associated with poor prognosis.

To determining the postmortem interval (PMI) through quantitative analysis of the DNA degradation of cell nucleus in human brain and spleen by using image analysis technique (IAT). The brain and spleen tissues from 32 cadavers with known PMI were collected, subjected to cell smear every 1 h within the first 5-36 h after death, stained by Feulgen-Van's staining, Three indices reflecting DNA in brain cells (astrocytes) and splenic lymphocytes, including integral optical density (IOD), average optical density (AOD), average gray (AG) were measured by employing the mage analysis instrument. The results showed that IOD and AOD declined and AG increased with the prolongation of dead time within 5-36 h. A correlation between the PMI and gray parameters (IOD, AOD and AG) was identified and the corresponding regression equation was obtained. The parameters (IOD, AOD and AG) were proved to be effective quantitative indicators for accurate estimation of PMI within 5-36 h after death.
Cell Nucleus/*pathology ; DNA Degradation, Necrotic ; Forensic Pathology ; Liver/*pathology ; Postmortem Changes ; Spleen/*pathology ; Time Factors

Objective To explore the relationship between T-cell acute lymphoblastic leukemia and the Notch signaling pathway.Methods Human T-cell acute lymphoblastic leukemia SupT1 cells were infected with the lentiviral vector made up specific Notch1-shRNA gene and nonspecific Notch1-shRNA gene.The inhibitive rate of SupT1 cells was detected by CCK-8.The rates of early apoptotic cells (Annexin V+/ 7-AAD-) and late apoptotic cells (Annexin V+/7-AAD+) were analyzed by flow cytometry and the expression levels of Notch1 receptor gene and downstream target genes were assessed by quantitative reverse transcription and polymerase chain reaction (QT-PCR).Results The cell inhibition rates of Notch1 interference group,control group and empty vector group at 96 h were 0.902±0.013,0,and 0.486±0.084,respectively,and it was increased obviously in Notch1 interference group (both P ＜ 0.05).The cell early apoptosis rates of the three groups were (15.27±0.31) ％,(5.57±0.25) ％,(5.80±0.20) ％,respectively.The cell early apoptosis rate of Notch1 interference group was increased obviously compared with the control group and empty vector group (both P ＜ 0.05).While the cell late apoptosis rates had no significant difference among the three groups (P ＞ 0.05).The mRNA expression levels of Notch1 receptor gene and its target genes (Hesl,c-myc,NF-κB) at 48 h,72 h and 96 h were higher than those in the control group and empty vector group (all P ＜ 0.05).Conclusions The specificity of Notch1-shRNA can effectively decrease the Notch1 mRNA expression,and reduce the expression level of downstream target genes.Notch1 cut can inhibit the proliferation of SupT1 cells,and promote the early apoptosis.

Objective To explore the frequencies of MPL exon 10 mutations in JAK2 V617F-negative myeloproliferative neoplasms patients. Methods MPLW515K/L was processed through allele-specific PGR combined with direct sequence analysis. The mutations of others MPL exon 10 were detected by single strand conformation polymorphism PGR (PCR-SSCP) combined with direct sequencing. Results Of the 103 patients with JAK2 V617F-negtive myeloproliferative neoplasms patients, 1 carried MPLW515K mutation (TGG→AAG)in PMF; 1 was found to have new mutation (CTGGTGATCGCT insert) in ET and have homozygous mutation. Conclusion JAK2 V617F-negtive myeloproliferative neoplasms patients carried additional mutations in addition to W515K/L mutations in MPL exon 10, but its frequency of mutation is low.

Objective To evaluate the value of immunophenotype in diagnosis of myelodysplastic syndrome (MDS).Methods The immunophenotype of bone marrow cells in 27 patients with MDS were detected by monoclonal antibody by flow cytometry.Results As the progression of the disease,CD34 positive cells gradually increased:refractory anemia/ring sideroblasts refractory anemia (RA/ AS) 7.43 %,refractory anemia with excess of blasts (RAEB) 36.81%,refractory anemia with excess of blasts transformed (RAEB-T)56.45 %,and the differences were statistically significant (P ＜0.05); the expressions of CD33+,CD13 and HLA-DR increased gradually,the expressions of CD14 and CD15 antigens gradually decreased,the difference of three groups was statistically significant (P ＜0.05),the differences between RA/RAS and RAEB-T,RAEB and RAEB-T were statistically significant (P ＜0.05); the expression of CD19 and CD10 decreased and the expression of CD7 increased (RA/RAS 2.63 %,RAEB 10.79 % and RAEB-T 11.00 %) with the progression of the disease,the difference of three groups was statistically significant (F =10.439,P ＜0.05),the differences between RA/RAS and RAEB,RA/RAS and RAEB-T were statistically significant (P ＜0.05).Conclusion The detection of immunophenotype of bone marrow cella in patients with MDS contributes to the diagnosis,classification and prognosis of MDS.

Objective To study the role of hematology analyzer in the preliminary diagnosis of hematological malignancies. Methods The blood routine results of 121 patients with initial hematological malignancies in the Second Hospital of Shanxi Medical University from October 2016 to February 2017 were analyzed by differential blood count (DIFF) scatter chart and alarm information of hematology analyzer SYSMEX XE-2100. Results Among 121 patients with hematological malignancies, 96 cases (79.3 %) had abnormality of DIFF scatter chart. The DIFF scatter chart of acute lymphoblastic leukemia, chronic lymphocytic leukemia, acute promyelocytic leukemia, and chronic myeloid leukemia had their own distinct characteristics, but the features of other types were not obvious. According to blood smear, blood cells analysis alarmed that the coincidence rates of primitive cells, immature granulocytes, nucleated erythroblasts and atypical lymphocytes were 87.7 % (50/57), 76.3 % (42/55), 33.3 % (19/57), 43.1 % (31/72) respectively. Conclusion The DIFF scatter chart of hematology analyzer and alarm information play very important roles in the diagnosis of hematological malignancies.

To determining the postmortem interval (PMI) through quantitative analysis of the DNA degradation of cell nucleus in human brain and spleen by using image analysis technique (IAT). The brain and spleen tissues from 32 cadavers with known PMI were collected, subjected to cell smear every 1 h within the first 5-36 h after death, stained by Feulgen-Van's staining, Three indices reflecting DNA in brain cells (astrocytes) and splenic lymphocytes, including integral optical density (IOD), average optical density (AOD), average gray (AG) were measured by employing the mage analysis instrument. The results showed that IOD and AOD declined and AG increased with the prolongation of dead time within 5-36 h. A correlation between the PMI and gray parameters (IOD,AOD and AG) was identified and the corresponding regression equation was obtained. The parameters (IOD,AOD and AG) were proved to be effective quantitative indicators for accurate estimation of PMI within 5-36 h after death.

Objective To explore the value of the plasma miR-193a-5p level on diagnosis and monitoring the response to treatment in acute myeloid leukemia (AML). Methods Peripheral blood samples were obtained from AML patients enrolled in hematology department of the Second Hospital of Shanxi Medical University from July 2015 to December 2015, including 30 de novo AML patients, 9 patients in complete remission (CR) and 6 patients in relapse. Peripheral blood samples from 15 healthy people were randomly choosed as the health control group. Plasma miR-193a-5p expression levels were detected by using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Results The plasma miR-193a-5p relative expression level of AML patients group [0.465 6 (0.103 1-5.000 2)] was obviously lower than that of health control group [0.766 1 (0.052 1-3.134 4)] (U= 122, P= 0.018 7). The plasma miR-193a-5p relative expression levels of de novo group and relapse AML group were significantly lower than those of CR group and health control group (P<0.05), and there was no significant difference between the CR group and health control group (U= 56, P= 0.511 9). No significant correlation was found between the plasma miR-193a-5p level and age, gender, blast percentage of the bone marrow, peripheral blood leukocyte count, platelet count, CD34+cells'percentage and so on. Conclusion The decreased plasma miR-193a-5p expression level can be served as a new and noninvasive biomarker for the evaluation of diagnosis and treatment in AML.

Objective To investigate the relevance between CD25 and common markers of immune phenotype in acute myeloid leukemia (AML), and the relationship between CD25 and FLT3-ITD gene. Methods The expression of mono-clonal antibodies, including, CD7, CD117, CD33, CD34, CD38, cyMPO, CD13, CD36, CD56, CD11b, CD19, CD16, CD14, CD64 and interleukin-2 three receptor chain CD25 (αchain), CD122 (β chain), CD132 (γ chain) were detected by immunofluorescence flow cytometry in 36 cases of AML bone marrow blast. The expression of every group of pa-tients with FLT3-ITD gene were detected by PCR. Results In 36 cases of AML patients, the rate of CD25+was 13.89%(5/36), 12.5% (1/8) in AML-M2 group, 18.18% (2/11) in AML-M4 group, 20% (2/10) in AML-M5 group, respectively, and expression rate of CD25+had been no statistically differences between those subgroups (P> 0.05). The percentage of bone marrow blasts which express CD7, CD117, CD33, CD34, CD38, cyMPO, CD13, CD36, CD56, CD11b, CD19, CD16, CD14, CD64, CD122, CD132 monoclonal antibodies has no significant difference between CD25+AML-M2 group and CD25-AML-M2 group, CD25+AML-M4 group and CD25-AML-M4 group, CD25+AML-M5 group and CD25-AML-M5 group, respectively (P> 0.05). Seven cases of AML patients with FLT3-ITD mutation, in five cases of CD25+AML pa-tients three case accompany FLT3-ITD+ mutation, the rates of FLT3-ITD+in CD25+AML was 60%, and higher than CD25-AML group (P <0.05). Conclusion (1)The expression rate of CD25 in AML patients is lower, and no significant correlation in patients in different AML FAB subtypes and common myeloid phenotype markers. (2)Since the FLT3-ITD mutation rates are higher in CD25+AML patients, there is important significance to clear the correlation between CD25 and FLT3-ITD gene.

Objective: To investigate the effect of biology and mTOR pathway activity of down-regulated TSC2 gene expression on U937 leukemia cells. Methods: Gene expression was down-regulated by lentivirus induced RNA interference on TSC2 high expressed U937 cell line; the proliferation, apoptosis and differentiation were detected by CCK-8 assay, colony formation assay and flow cytometry; the gene expression level and protein kinase activity were detected by qRT-PCR and Western blot. Results: Down-regulated expression of TSC2 gene promoted U937 cell proliferation and colony formation ability (P<0.05) . The proportion in G₀/G₁ phase of TSC2 down-regulated U937 cell was much lower than that of the control cells [ (52.53±3.75) % vs (75.10±4.33) %, t=6.829, P=0.002], the S phase [ (22.43±1.00) % vs (15.47±1.20) %, t=-5.581, P=0.019] and G₂/M phase [ (25.03±4.34) % vs (14.33±0.91) %, t=-5.413, P=0.013] was remarkably higher than that of the control cells (P<0.05) . There were no statistically significant differences in cell apoptosis and differentiation (P>0.05) . Down-regulation of TSC2 led to the increased activity of mTOR, 4EBP1 and S6K1, but did not influence the activity of AKT. The expressions of proliferation related cyclinD1, c-myc and PTEN were also up-regulated after TSC2 silenced, but the expressions of P27KIP and BCL-XL were not changed. Conclusion Downregulation of TSC2 could promote the proliferation of U937 cells through up-regulation of mTOR activity.