[The Diagnostic Value of Peripheral Blood Cell Parameters for Early Recognition of Myelodysplastic Syndrome].

OBJECTIVE: To evaluate the diagnostic value of peripheral blood cell parameters for early recognition of myelodysplastic syndrome (MDS) patients. METHODS: The clinical and laboratory data of 86 patients with MDS and 72 patients with non-malignant clonal anemia treated in first diagnosed in the Second Hospital of Hebei Medical University from January 1, 2015 to December 31, 2017 was retrospectively analyzed. The peripheral blood cell parameters of the patients in two groups were analyzed, generated the receiver operator characteristic curve (ROC curve) from the statistically significant parameters, the binary logistic model was build to calculate and compare the area under the ROC curve (AUC) combined with multiple indicators and individual indicators, sensitivity, specificity, positive and negative likelihood ratio, and diagnostic accuracy, the diagnostic efficacy of the patients was analyzed. RESULTS: Compared with patients in the non-malignant clonal anemia group ,white blood cell count (WBC), neutrophil percentage (NE%), eosinophil percentage (E%), eosinophil absolute value (E#), platelet count (PLT), platelet specific volume (PCT%) in the MDS patients were significantly reduced; while percentage of lymphocytes (LY%), basophilic percentage (B%), and the width of platelet distribution (PDW) significantly increased. The several ROC curves with the above indicators were established, which showed that AUCLY%%=0.718 (P=0.040); AUCPDW=0.674 (P=0.044); AUCB%=0.650 (P=0.044) were >0.5. After established a binary logistic regression model, the AUCPRE-4 obtained by combining the three indicators of LY%, PDW and B% was 0.777 (P=0.037), which was significantly higher than the AUC of any indicator alone. When the sensitivity was 77.91% and the specificity was 61.11%, the corresponding threshold value was 0.47, the positive likelihood ratio was 2.00, the negative likelihood ratio was 0.36, and the case ratio of correct classification was 54.40%. CONCLUSION: PDW, B% and LY% in peripheral blood cell parameters have certain diagnostic value for early recognition of MDS.

Arsenic trioxide-induced cell apoptosis and cell cycle arrest are potentiated by 1,25-dihydroxyvitamin D3 in human leukemia K562 cells.

The interaction between 1,25-dihydroxyvitamin [1,25(OH)2D3] and vitamin D receptor (VDR) plays a critical role in regulating cell proliferation and programmed cell death. The present study aimed to investigate the effects of 1,25(OH)2D3 in combination with arsenic trioxide (As2O3) on the proliferation and cell cycle of a K562 leukemia cell line. K562 cells were treated with 100 nM 1,25(OH)2D3, 2.5 µM As2O3, and 100 nM 1,25(OH)2D3 combined with 2.5 µM As2O3. Cell proliferation was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt/phenazine ethosulfate method. Cell cycle progression and apoptosis were detected by flow cytometry. The expression levels of genes associated with the cell cycle and apoptosis were analyzed by reverse transcription-quantitative PCR and western blotting analyses. The present findings indicated that combined treatment of 1,25(OH)2D3 and As2O3 led to a significant increase in cytotoxicity, apoptotic cell death and G1 cell cycle arrest when compared to those treated with 1,25(OH)2D3 or As2O3 alone. The downregulation of the Bax/Bcl-2 ratio and decreased survivin expression may be involved in combined treatment-mediated apoptosis. G0/G1 cell cycle arrest induced by combined treatment was associated with the activation of p21 and p27. In addition, the increased expression of VDR was found to participate in the anticancer effect of combination treatment. The data suggested that the combination of 1, 25-(OH)2D3 and As2O3 had clear synergistic effects on the inhibition of K562 cell proliferation, which could provide a novel therapeutic approach for the treatment of acute myeloid leukemia.

[Effects of As2O3 on the Proliferation, Differentiation and Apoptosis of HL-60 Cells and Its Related Mechanisms].

OBJECTIVE: To investigate the effects of arsenic trioxide (As2O3) on the proliferation, differentiation and apoptosis of HL-60 cells in vitro and explore the underlying mechanisms. METHODS: After HL-60 cells were treated with different concentration of As2O3, the cell proliferation was determined by MTS/PES method, the differentiation state was detected by the nitroblue tetrazolium (NBT) reduction test; flow cytometry was used to analyze the apoptosis and expression of CD11b. In addition, SYBR Green real-time RT-PCR was used to measure the mRNA levels of C-FES, BCL-2, BAX, survivin , P21 and P27. RESULTS: As2O3 could obviously inhibit the proliferation of HL-60 cells, and the effect was in dose- and time-dependent manners (r=-0.967; r=-0.954). Low concentration (0.1, 0.5 and 1.0 µmol/L) of As2O3 could significantly promote the differentiation of HL-60 cells, the cells exhibited a higher NBT-reducing ability and expressed far more CD11b antigens. High concentration (2.5 and 5.0 µmol/L) of As2O3 induced HL-60 cell apoptosis, but the ability of promoting differentiation decreased. The expression of C-FES mRNA significantly increased after being treated with As2O3 at the concentrations 1.0 and 5.0 µmol/L, and the former is more obvious, which confirmed that C-FES mRNA level paralleled the cell differentiation degree. Also, the expression of BCL-2 and survivin significantly decreased, while the expression of BAX, P21 and P27 was significantly upregulated in HL-60 cells after being treated with 5.0 µmol/L As2O3. CONCLUSION: As2O3 can significantly suppress cell proliferation, promote the differentiation and induce the apoptosis in HL-60 cells, and the mechanism of As2O3 anti-tumor activity may be involved in the regulation of C-FES, cell cycle and apoptosis-related genes.

Efficacy and Safety of Lenalidomide in the Treatment of Multiple Myeloma: A Systematic Review and Meta-analysis of Randomized Controlled Trials.

BACKGROUND: Lenalidomide has emerged as an important treatment for patients with multiple myeloma (MM). However, its role in the management of MM is still controversial and requires further clarification. The aim of this study was to evaluate efficacy and safety of lenalidomide for MM using a meta-analysis. METHODS: We searched the electronic databases including: PubMed, EMBASE and the Cochrane Center Register of Controlled Trials. Seven randomized clinical trials were identified, which included a total of 2357 patients with MM who received lenalidomide-containing, noncontaining lenalidomide regimens or placebo as induction therapy or maintenance therapy. The outcomes included overall response (OR) rate, complete response (CR) rate, 3-year progression-free survival (PFS) rate, 3-year overall survival (OS) rate, and different types of treatment-related adverse events. We calculated the risk ratios (RRs) as well as their 95% confidence intervals of these outcomes and pooled the results using RevMan 5.2 software. RESULTS: For patients with previously untreated MM, OR rate and CR rate was significantly higher in lenalidomide-containing group than the control group. For relapsed or refractory MM patients, lenalidomide-containing regimens significantly improved the OR rate, CR rate, 3-year PFS rate and 3-year OS rate. With regard to MM patients after autologous stem cell transplantation, lenalidomide maintenance therapy significantly improved 3-year PFS rate but did not result in improved 3-year OS rate. In terms of toxicities, lenalidomide therapy has a higher rate of Grade 3-4 grade cytopenias, infection, deep-vein thrombosis, and diarrhea. Furthermore, the incidence of second primary malignancies was significantly higher in the lenalidomide group. CONCLUSIONS: The lenalidomide-containing regimens as induction therapy clearly increased response rates and improved intervals of survival with acceptable toxicity rates for patients with MM. However, when physicians choose to use the lenalidomide as maintenance therapy, whether the benefits outweigh the risks should be taken into account.

Quantitative detection of the human cervical cancer oncogene for monitoring the minimal residual disease in acute leukemia.

The human cervical cancer oncogene (HCCR) has been shown to be over-expressed in some solid tumors, and its function is involved in negative regulation of p53 tumor suppressor gene. However, the roles of HCCR in leukemia remain unclear. The present study is to investigate whether the expression levels of HCCR mRNA are associated with clinical prognosis in patients with acute leukemia (AL) and to explore the potential use as a biomarker for monitoring minimal residual disease (MRD) in AL. The mRNA levels of HCCR1 and HCCR2 were quantified by real-time reverse transcription polymerase chain reaction in bone marrow samples from 80 adult de novo AL patients and 20 normal healthy donors. The expressions of HCCR1 and HCCR2 were significantly higher in patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) than those in healthy donors (P < 0.01), but there was no significant difference between AML and ALL (P > 0.05). Besides white blood cell count, we did not find any significant correlation between HCCR expression and clinical characteristics, such as age, sex, CD34 antigen expression, and response to chemotherapy. HCCR was monitored in 12 cases during remission and/or relapse. Significant reductions of both HCCR1 and HCCR2 mRNA levels were observed in patients who had achieved complete remission after chemotherapy but not in patients with non-responsive. However, an increased HCCR expression was detected in these patients who relapsed. Our findings suggest that HCCR gene is over-expressed in AL patients and may be as a useful biomarker for monitoring MRD in AL.

Compound heterozygous hemophilia A in a female patient and the identification of a novel missense mutation, p.Met1093Ile.

Hemophilia A (HA) in females is rare. Female HA cases are often misdiagnosed as acquired HA (AHA) or as von Willebrand disease type 2N (vWD-2N). Here, we report the case of a 37-year-old female HA patient with a moderate factor VIII (FVIII) deficiency. The patient had no personal or family history of bleeding disorders, but presented with heavy uterine bleeding following surgery to remove an intrauterine device. IgG inhibitory antibodies against FVIII were undetected. A compound heterozygote mutation of the FVIII gene (F8) was found in this patient. The p.Val502Asp mutation, which has been reported previously, affects A2 domain function. A novel missense point mutation, p.Met1093Ile, was identified in the B domain. The compound heterozygote mutations in F8, p.Val502Asp and p.Met1093Ile, caused HA in this female patient, with a moderate phenotype.

[Construct Mxi1 mutation gene expression vector from leukemia cell line KG1].

OBJECTIVE: To construct the eukaryotic expression vector for Max interacting protein 1 (Mxi1). METHODS: The full length cDNA of Mxi1 gene obtained from fetal lymphocyte and KG1 cells were inserted into plasmid pDs-red2-N1 respectively to generate pDs-red2-N1/Mri1 (wild/mutation type). Then the recombinant vector was transfected into Cos-7 cells via liposome. 48 hours post transfection, mRNA of Mri1 gene was detected by RT-PCR and Mxi1 protein expression was detected by flow cytometry and fluorescence microscope in the Cos-7 cells. RESULTS: The eukaryotic expression vector of Mxi1 was constructed and transfected into eukaryotic cells successfully. The expression of red fluorescence protein in the transfected Cos-7 cells was observed under fluorescence microscope which implied the expression of Mxi1. The transfect efficiency of both wild and mutation type were in a high level in 3 days after transfected, which lasted to 6 d. RT-PCR amplified the total RNA extracted from the transfected Cos-7 cells could find increased mRNA level of Mxi1 gene. CONCLUSION: We successfully constructed the eukaryote expression vector for Mri1 gene; Cos-7 cells transfected with the vector via liposome could express Mxi1 protein. These could be useful for the further study of the Myc gene modulation.

[Expression of c-fes gene in leukemia cells and its clinical significance].

This study was purposed to investigate the expression of c-fes gene in leukemia patients and its clinical significance. The expression of c-fes mRNA in bone marrow cells from 121 cases of acute and chronic leukemia patients, and the expression of c-fes mRNA in peripheral blood mononuclear cells of 20 normal persons were detected by real time-quantitative reverse transcription polymerase chain reaction (RQ-PCR). The results showed that the level of c-fes mRNA in AML patients was higher than that in normal controls [(48.017 +/- 57.170) x 10(-3) vs (0.152 +/- 0.398) x 10(-3)] (p < 0.0001); but there was no significant differences of level of c-fes mRNA between samples of ALL and normal controls(0.047 +/- 0.068) x 10(-3) vs(0.152 +/- 0.398) x 10(-3) (p>0.05); the level of c-fes mRNA in CML patients was higher than that in normal persons (21.605 +/- 24.818) x 10(-3) vs (0.152 +/- 0.398) x 10(-3) (p < 0.0001). The positive expression rate of c-fes gene in CML-CP patients (80%) was higher than that in CML-AP patients (66.7%) and CML-BP (28.6%) patients. In AML patients, c-fes gene was expressed higher in M(2) (80.77%) and M(3) (92.86%) patients. The remission rate of AML (except M(3))patients who had expression of c-fes gene was 81.08%, which was higher than that of patients with no expression of c-fes gene (40.00%). It is concluded that c-fes gene expression was found in myeloid leukemias, whereas low or no expression in lymphocytic leukemias. The differentiation of myelocytic cells may be related to c-fes gene. All AML (except M(3))patients with high level of c-fes mRNA may get good prognosis.

[The expression and clinical significance of survivin gene in leukemia].

OBJECTIVE: To investigate the expression of survivin in leukemia and the prognostic significance in acute leukemia (AL). METHODS: The expression of survivin mRNA was measured in 105 AL and 21 chronic myelogenous leukemia (CML) patients with semi-quantity reverse transcription (RT)-PCR. 15 adults were tested as normal control (NC) and K562, NB4, Kg-1alpha, HL-60 cell lines were also tested as positive control. The cell cycle distribution in AL was measured with flow cytometry (FCM) to analyze the relationship between the level of survivin and cell proliferation. RESULTS: The expression of survivin in de novo AL (0.525 +/- 0.460) was higher than that in NC (0.101 +/- 0.187), while it decreased in complete remission (CR) patients (0.280 +/- 0.095). In replased patients (0.935 +/- 0.343), the expression of survivin increased again. There was no difference of the expression between chronic phase of CML (0.279 +/- 0.112) and NC, but in acute phase of CML (0.653 +/- 0.236), the expression was higher than that in NC. The cases with higher level of survivin had significantly higher proliferation indices (mean PI 9.682) than those (mean PI 6.899). In AL patients, the CR rate of survivin positive cases was lower than that of survivin negative cases. CONCLUSIONS: The PI in survivin positive patients was significantly higher than that in survivin negative indicating its impact on proliferation and cleavage. Abnormal expression of survivin genes was related to pathogenesis and progression of AL and it can serve as a marker of relapse and poor prognosis in AL. Overexpression of survivin is a risk factor for CML.

[Expression and significance of X-linked inhibitor of apoptosis protein and its antagonized proteins in acute leukemia].

To investigate the expression and significance of X-linked inhibitor of apoptosis protein (XIAP) and XIAP-associated factor 1 (XAF1) in acute leukemia, the expression of XIAP, XAF1, Smac, and HtrA2 mRNA in the bone marrow aspirates from 87 newly diagnosed AL patients, 23 patients in remission, 6 patients in relapse, and 17 normal controls were detected by means of reverse transcriptase polymerase chain reaction (RT-PCR), and their relationship with clinical therapeutic efficiency was analyzed. The results showed that the expression level of XIAP mRNA in newly diagnosed AL patients (0.990 +/- 0.337) was significantly higher than that in normal controls (0.395 +/- 0.148) (P < 0.01); the positive rate and expression level of XAF1 mRNA in newly diagnosed AL patients (56.32%, 0.246 +/- 0.267) were significantly lower than that in normal controls (100%, 0.964 +/- 0.387) (P < 0.01). In 69 out of 87 newly diagnosed AL patients, efficacy remained evaluable. AL patients with high level of XIAP achieved a lower complete remission (CR) rate than patients with low level of XIAP (54.55% and 86.11%, respectively) (P < 0.01). XAF1 positive patients achieved a higher CR rate than XAF1 negative patients (86.84% and 51.61%, respectively) (P < 0.01). It is concluded that the overexpression of XIAP and negativity of XAF1 may be two adverse prognostic factors in AL patients.

Expressions of cyclin E2 and survivin in acute leukemia and their correlation.

Cyclin E2 is present in solid tumors, while its expression and clinical value in acute leukemia is unknown. This study was aimed to investigate the expression of cyclin E2 and survivin gene in bone marrow cells from patients with acute leukemia and their relationship. Reverse transcription polymerase chain reaction was used for detection of the expression of cyclin E2 and survivin mRNA in 84 adult patients with acute leukemia which included 16 cases of relapse, 60 cases of de novo acute leukemia, 8 cases of continuously complete remission, and 20 normal persons as controls. The results showed that (1) positive expression of cyclin E2 (70.24%) in acute leukemia patients was significantly higher than that (0%) in controls, positive expression of survivin (72.62%) in acute leukemia patients was higher than that (30%) in control. (2) the expression of cyclin E2 positively correlated with that of survivin in acute leukemia patients. (3) remission rate in cyclin E2-positive patients (55.81%) was lower than that (88.24%) in cyclin E2-negative patients, the rate of cyclin E2 expression in relapse group was the highest among the three groups; while that in continuously complete remission group was the lowest among the three groups. (4) positive rate of cyclin E2 expression (59.32%) in patients with acute myelocytic leukemia was lower than that (96%) in patients with acute lymphocytic leukemia, no correlation between cyclin E2 expression and white blood cell counts of patients was found. It is concluded that the overexpression of cyclin E2 has been confirmed for the first time to positively correlate with the expression of the survivin in acute leukemia patients, and implicate the poor prognosis. Cyclin E2 may be used as a marker for examination of minimal residual disease.

[Relationship between resistance to chemotherapy and expression of breast cancer resistance protein (BCRP) gene in patients with acute leukemia].

In order to investigate the relationship between the expression of breast cancer resistance protein (BCRP) gene and drug resistance in patients with acute leukemia (AL), semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the expression of BCRP mRNA in AL patients and 15 normal subjects. Beta(2)-microglobin (beta(2)-MG) was used as positive reference. BCRP/beta(2)-MG ratio >or= 0.5 was defined as BCRP mRNA positive. The results showed that the positive percentage of BCRP gene expression in newly diagnosed group was 37.6%. The first complete remission rate was 79.3% and 31.6% in BCRP mRNA negative and BCRP mRNA positive patients respectively. The difference was significant (P = 0.001). The expression levels of BCRP mRNA in drug resistance group and drug sensitive group were 0.962 +/- 0.426 and 0.315 +/- 0.296 respectively (P = 0.0001). The expression level of BCRP mRNA in relapsed/refractory group was significantly higher than that in newly diagnosed group (P = 0.0025). The expression level of BCRP gene in normal individuals and long-term survival group was very low and correlated with FAB subtypes. It is concluded that high expression of BCRP mRNA leads to clinical drug resistance and is an unfavorable factor for prognosis of AL patients except acute promyelocytic leukemia.

[Blastic natural killer cell leukemia--one case report and review of literature].

OBJECTIVE: To report a case of blastic natural killer cell leukemia with an aggressive clinical course. METHODS: The characteristics of blastic NK cell leukemia and its treatment were discussed with review of literatures. RESULTS: After combination chemotherapy and spinal cord segmental radiotherapy, the patient entered hematological remission, but the extramedullary lesion remained unchanged. CONCLUSION: Blastic NK cell leukemia has an aggressive clinical course with poor response to treatment and unfavorable prognosis.

[Expression of survivin, Fas, bcl-2 and bax in bone marrow cells from acute myeloid leukemia patients and its clinical significance].

UNLABELLED: To study the clinical significance of the expression of antiapoptosis gene, survivin and bcl-2, and proapoptosis gene, Fas and bax, in acute myeloid leukemia (AML), RT-PCR was used to examine the expression of survivin and flow cytometry (FCM) to detect the expression of Fas, bcl-2, bax and bcl-2/bax ratio in 68 cases of AML. The results demonstrated that: (1) The positivity of survivin mRNA expression was significantly higher in AML compared to control (70.6% vs 30%, P < 0.05). (2) The expression of Fas and bcl-2 in AML before treatment was significantly higher than that in control (P < 0.001), but the bax expression did not (P > 0.05). (3) The survivin-positive AML cases showed a significantly lower Fas and higher bcl-2 expression in comparison with survivin-negative ones (P < 0.01 and P < 0.001, respectively), but the bax did not (P > 0.01). (4) Survivin-positive AML cases had a lower CR rate as compared with survivin-negative cases (64.6% vs 90%, P < 0.05). (5) The survivin-positive CR cases showed a decreased expression of Fas and bcl-2 after treatment in comparison with pretreatment expression (P < 0.001), but the bax expression remained unchanged before and after therapy. The survivin-positive NR cases showed a significantly decreased Fas and increased bcl-2 expression as compared with pretreatment expression (P < 0.001). bcl-2/bax ratio was also significantly higher in NR cases. IN CONCLUSION: 70.6% AML cases showed positive for survivin expression with a lower CR rate, the survivin-positive AML showed a low Fas with high bcl-2 expression and bcl-2/bax ratio as compared to the survivin-negative cases.