ABSTRACT
Objective:To investigate the predictive efficacy of the established prognostic nomogram of rituximab in treatment of diffuse large B-cell lymphoma (DLBCL) patients with bone marrow infiltration.Methods:The clinicopathological data of 71 DLBCL patients with bone marrow infiltration who received first-line treatment with rituximab between January 2014 and June 2016 in Shanxi Province Cancer Hospital were retrospectively analyzed. Progression-free survival (PFS) analysis was performed by using Kaplan-Meier method, and influencing factors of PFS were analyzed by using univariate and multivariate Cox proportional hazards models. The nomogram was drawn with R software based on independent influencing factors of PFS from Cox regression analysis. Receiver operating characteristic (ROC) curve was applied to evaluate the effects of nomogram models predicting the PFS of patients; Bootstrap method was used for internal validation of the model. A nomogram calibration curve was plotted to compare the consistency between the nomogram model prediction and the actual PFS.Results:The median follow-up time of all patients was 48 months (12-84 months), and the 3-year and 5-year PFS rates were 39.44% and 26.76%, respectively. Age > 60 years ( HR = 1.593, 95% CI 1.379-1.840, P < 0.001), Ann-Arbor staging Ⅲ-Ⅳ ( HR = 1.444, 95% CI 1.092-1.910, P = 0.010), international prognostic index (IPI) score 3-5 ( HR = 1.648, 95% CI 1.249-2.333, P < 0.001), complicated with type 2 diabetes ( HR = 5.880, 95% CI 1.645-21.023, P = 0.006) were independent influencing factors of PFS in DLBCL patients with bone marrow infiltration. The independent influencing factors of PFS were included to establish the prognostic nomogram model. Bootstrap method internal validation showed that the consistency index of the prediction model was 0.71 (95% CI 0.69-0.78), and the ROC curve showed that the area under the curve (AUC) of 3-year PFS predicted by nomogram model was 0.708, 5-year PFS predicted by nomogram model was 0.716, indicating that nomogram model had a good degree of differentiation; and the calibration curve results showed that the 3-year and 5-year PFS rates predicted by nomogram model had a good consistency with the actual 3-year and 5-year PFS rates. Conclusions:The nomogram model constructed by age, Ann-Arbor staging, IPI score, complicated with or without type 2 diabetes could be used to predict the prognosis of DLBCL patients with bone marrow infiltration treated with rituximab, which is helpful for clinicians to implement treatment strategies.
ABSTRACT
Objective To develop a real-time quantitative PCR method with TaqMan probe to analysis the lineage-specific chimerism based on single nucleotide polymorphisms (SNP).Methods CD3 positive and CD15 positive cells were separated by magnetic cell sorting system from cord blood,and a quantitative method were established using real-time quantitative PCR and SNP.Detect the artificial chimerism and mark the standard curves.The reaction system was optimized,and the sensitivity and specificity were evaluated.Results Separation purity of blood cells by magnetic cell sorting system was up to 94 ％-97 ％.Discrimination between donor and recipient was possible.Dilution experiments of the mock chimerism sample revealed that Ct values correlated linearly with the logarithm of recipient/donor DNA fraction (r ＞ 0.98),and the limit of detection for a minor DNA percentage was under 0.1％,and the specificity was also good.Quantitative analysis of 4 clinical cases in same period were made by real-time fluorescence quantitative SNP-PCR,the fitted rate was 87.50 ％ based on the chimera standard curve calculated for 1 case.Conclusion The method shows high sensitivity and specificity,and will be used to quantify the lineagespecific chimerism after allogeneic hematopoietic stem cell transplantation.
ABSTRACT
Objective To investigate ATM deletion [del (ATM)] in chronic lymphocytic leukemia (CLL) and study its correlation with the clinical stage. Methods Spectrum Orange~(TM) labeled sequence specific DNA probe for ATM locus on 11q22.3 and fluorescence in Situ hybridization (FISH) was used to examine del (ATM) in 28 newly diagnose patients with CLL. FISH analysis were performed on bone marrow smears. Clinical staging was done according to Binet Method.Fisher exact propability was used to study the relations between del (ATM) and clinical feature. Results 4 out of the 28 cases were found with deletion of ATM. The incidence of del (ATM) in BinetA, BinetB and BinetC was 1/9 (11.1 %), 1/8 (12.5 %), 2/11 (18.2 %), respectively. Fisher exact propability showed that deletion of ATM was not associated with its clinical feature. Conclusion Application of FISH on archival bone marrow smears is a simple, liable method, and can be readly used to retrospective study of clonal blood system diseases. Deletion of ATM was common cytogenetic changes in CLL patients.And the significance of del (ATM) in the prognosis of CLL in China needs to be further investigated.
ABSTRACT
Objective To apply fluo-rescencc-activated cell sorting(FACS) in sorting T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+), which establish the separating method of series of cells from human peripheral blood, so that the scientific research and clinical research could be carried out were specifically. Methods 10 cases of normal peripheral blood were collected and T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) were stained with florescence conjugated antibodies. The positive cells were sorted by FACS. Results Before sorting the peripheral blood, the proportion of the T lymphocyte(CD_3~+) and granulocyte (CD_(15)~+) in the leukocyte is 48.8 % and 30.8 %; after sorting by FACS, the purity of T lymphocytes (CD_3~+) is up to 98 % and the recovery is about 95 %; the purity of granulocyte (CD_(15)~+) is up to 97 % and the recovery is about 96 %. Conclusion The FACS could Mlow us to quickly sort T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) with higher recovery and higher purity from the peripheral blood.
ABSTRACT
Objective To investigate the correlation and significance of cyclooxygenase-2 (COX-2)and the breast cancer resistance protein (BCRP) expressions in non-Hodgkin lymphoma(NHL). Methods Ten patients with reactive lymph nodes (RLN) were considered as a control group. Compared with β-actin as internal control, the BCRP mRNA expressions of 61 NHL samples and 10 lymph tissues in control group were detected by semi-quantitative revere transcription polymerase chain reaction (RT-PCR) assay, while the expressions of COX-2 protein of the above specimens were detected by SP immunohistochemistry. Results The positive rates of COX-2 and BCRP in NHL were 50.8 %(31/61) and 45.8 % (28/61), respectively , and were higher than those in the control group (P ＜0.05). The expression of COX-2 was statistically positive correlated to that of BCRP (X2 =8.795, r=0.355, P＜0.05). The expressions of COX-2 and BCRP were not correlated to clinical and pathological factors, such as age, sex, IPI, LDH, β2-microglobulin level and Ann Arbor stage, however, the expression of BCRP was statistically correlated to chemotherapy efficacy.Conclusion BCRP may be involved in multi-drug resistance (MDR) of NHL, so it may contribute to the assessment of chemotherapy and prediction of NHL. Since there is a strong correlation between COX-2 expression and MDR in NHL, the application of COX-2 inhibitors may enhance sensitivity of chemotherapy.
ABSTRACT
Objective To study the over-expression and clinical implications of the oncogene MDM2 in acute leukemia (AL). Methods The expression of MDM2 gene in 100 patients with newly diagnosed and relapse or refractory AL and 20 healthy as control was measured by relative quantitative reverse transcriptase polymerase chain reaction (RT-PCR),then the results was measured by χ2-test,t-test and one-way ANOVA to compare expession positive rate and intensity of MDM2. Results Among 100 patients,fifty-eight had the high expression of MDM2 gene (58 %). The expression level of MDM2 gene in patients was higher than that of health controls(P ＜0.05). The expression positive rate of MDM2 is higher in poor outcome group (67.9 %,19/28)than that in general outcome group (33.9%,19/56) (P＜0.05). Conclusion Our results suggest that the expression of MDM2 gene plays an important role in the pathogenesis and poor outcome of AL.