Clinical significances of CD4/CD8 ratio and neutrophil-to-lymphocyte ratio in patients with multiple myeloma / 白血病·淋巴瘤

Objective:To investigate the clinical significances of CD4/CD8 ratio and neutrophil-to-lymphocyte ratio (NLR) in patients with multiple myeloma (MM).Methods:The clinical data of 124 MM patients in the Third Affiliated Hospital of Soochow University from December 2002 to April 2017 were retrospectively analyzed, and 31 healthy people were chosen as the controls. Peripheral blood T lymphocyte subsets were detected by using flow cytometry, and the correlations between CD4/CD8 ration and related clinical indicators were also investigated. All MM patients were divided into the high NLR group and the low NLR group according to the media of NLR, and the correlation of them with related clinical indicators, chromosome karyotype, overall survival (OS) and progression-free survival (PFS) was also compared.Results:Compared with the healthy control group, the proportion of CD4 + T cells [(35.28±6.58)% vs. (31.85±6.76)%, t = -2.067, P = 0.043], absolute value of NK cells [0.22×10 9/L (0.13×10 9/L-0.59×10 9/L) vs. 0.17×10 9/L (0.00×10 9/L-0.42×10 9/L), Z = -2.614, P = 0.009] and CD4/CD8 ratio [0.97 (0.50-2.69) vs. 0.81 (0.30-1.28), Z = -2.253, P = 0.024] was decreased, respectively. The proportion of CD8 + cells was increased [(36.93±7.38)% vs. (40.50±6.50)%, t = 2.074, P = 0.042] in MM group. The hemoglobin level of CD4/CD8 ratio ≥0.94 group was higher than that of CD4/CD8 ratio <0.94 [(98.89±21.35) g/L vs.(80.60±23.23) g/L, t = -2.066, P = 0.047]. Compared with the healthy control group, NLR was increased in MM group [1.54 (1.10-3.23) vs. 1.95 (0.29-12.70), Z = -2.384, P = 0.017]. Compared with the low NLR group (<1.95), serum β 2-microglobulin [4.56 mg/L (1.63-12.60 mg/L) vs. 6.17 mg/L (1.58-67.50 mg/L), Z = -2.586, P = 0.010] and serum creatinine [84.5 μmol/L (43.0-376.5 μmol/L) vs. 113.0 μmol/L (46.5-754.0 μmol/L), Z = -3.866, P < 0.001] was increased in the high NLR group for MM patients. The proportion of the male patients, β 2-microglobulin > 5.5 mg/L, serum creatinine > 177 μmol/L, stage Ⅲ of international staging system (ISS) in the high NLR group was higher than that in the low NLR group (all P < 0.05), and there was no statistically significant difference in the composition of chromosome karyotype (all P > 0.05). The median OS time in the low NLR group was longer than that in the high NLR group [30 months (20-40 months) vs. 17 months (7-27 months), χ 2 = 4.519, P = 0.034], and there was no statistically significant difference in the PFS of both groups ( P > 0.05). Multivariate Cox analysis demonstrated that the age, corrected serum calcium, serum creatinine, lactic dehydrogenase were the independent influencing factors of OS in MM (all P < 0.05), while NLR wasn′t an independent influencing factor of OS in MM ( P = 0.513). Conclusions:CD4/CD8 ratio is decreased and immune dysfunction occurs in MM patients. MM patients with high NLR have a shorter OS time.

Proliferative and apoptotic effects of simvastatin in combination with cytosine arabinoside on K562 cells / 白血病·淋巴瘤

Objective To investigate the effect of simvastatin (SV) in combination with cytosine arabinoside (ARA-C) on the proliferation and apoptosis of K562 cells. Methods Human K562 cells were incubated with SV and cytosine arabinoside alone or in combination and K562 cells without any treatment were taken as normal control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detections. Morphological changes by Wright stain were performed. MTT method was used to assay the growth inhibition rate and cytoflowmetry was used to detect the early stage apoptosis ratio and cell necrosis ratio. Results Compared with Ara-C group and SV group, cells in the group treated with SV combined with Ara-C showed obvious karyopyknosis,apoptosis bodies formation and significant cell growth inhibition, which were positively correlated with culture time. Combination of 15 μmol/L SV and Ara-C showed the most significant cell growth inhibition with a inhibition rate of (72±1) % at 72 h of culture, as was significantly higher than that of 15 μmol/L SV group (45±2) % and 20 μmol/L Ara-C group (44±0) % (P ＜0.01),furthermore, combination of 15 μmol/L simvastatin and Ara-C showed synergistic inhibition with Q value of 1.24 and 1.19 at 24 h and 48 h in each. The apoptosis rates at early stage (AnnexinV) detected by flow cytometry in 20 μmol/L, 15 μmol/L and 10 μmol/L SV treated K562 cells were significantly higher than that in normal K562 cells (P ＜0.01), as were positively correlated with culture time and SV dose (P ＜0.05). There were no significant difference of early apoptosis rate between the 20 μmol/L SV and 15 μmol/L SV groups (P ＞0.05), yet the very two were both higher than that of 10 μmol/L SV group (P ＜0.05). There were no statistic differences of late apoptosis rate (PI) amongdifferent treated groups (P ＞0.05). Conclusion SV inhibited K562 cell proliferation and induced cell apoptosis in vitro, and combination of SV and Ara-C exhibited obvious synergistic inhibition and apoptosis, which may increase the sensitivity of K562 cell to chemotherapy. SV at 15 μmol/L may be the best concentration for K562 cells in vitro.

Correlation between translocation of 14q32 and deletions of 13q14 in multiple myeloma / 白血病·淋巴瘤

Objective To investigate the common chromosome abnormalities in the patients with multiple myeloma and the relationships of cytogenetic abnormalities and clinical features. Methods The interphase fluorescence in situ hybridization (I-FISH) analysis method was designed to detect RB1-/13q14-and 14q32 rearrangements in 49 MM patients. The statistic value of its effect on clinical features were determined. Results FISH disclosed 14q32 translocations in 26 of the 40 (53.1%) patients. 25 out of the 49 (51.02 %) cases were found with deletion of chromosome 13q14 included del(RB1) in 9 (18.4 %) and del(13q14.3) in 18 (36.7 %). 13q14 deletion and 14q32 translocation were simultaneously observed in 18 (36.7 %) cases. Spearman correlation analysis were found associated of 14q32 rearrangement with the percentage of plasma cells in bone marrow (r=0.316, P=0.27). Conclusion The frequency of 13q14 deletion and 14q32 gene translocation in multiple myeloma are high. There is a significant correlation between the presence of 14q32 translocations and chromosome 13 abnormalities in MM patients. The percentage of 14q32 translocation in plasma cells was increased significantly. The 14q32 translocation is an independent prognostic factor.

Effects of simvastatin on differentiation and apoptosis of human promyelocytic leukemia cell line NB4 / 白血病·淋巴瘤

Objective To investigate the effects of simvastatin (SV) on the proliferation,differentiation and apoptosis of human promyelocytic leukemia cell line NB4.Methods NB4 cells were incubated with SV at different concentration with or without all-trans retinoic acid (ATRA),and NB4 cells without any treatment were taken as normal control.Cells of different groups were collected at 24 h,48 h and 72 h after incubation for further detection.Morphological changes by Wright stain were performed.MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the surface CD11b expression levels,the early stage apoptosis ratio and cell necrosis ratio.Results Treated with 15 μ mol/L SV,10 μ mol/L SV and 5 μ mol/L SV respectively,with the NB4 cells growth,the cell inhibition rates gradually increased (F =7.15,P =0.000),as well as CD11b expression levels (F =3.41,P =0.014) and AnnexinVexpression levels (F =43.38,P =0.000).Furthermore the NB4 cells treated with 15 μ mol/L SV exhibited the most significant changes with cell inhibition rate of 0.96±0.02,CD11b expression level increased to (62.41±6.37) ％ and AnnexinV expression level increased to (87.38±2.94) ％ after 72 h incubation.Combination of 15 μmol/L SV with 0.5 μmol/L ATRA displayed obvious interaction for increasing CD11b expression levels (F =4.093,P =0.025),while no significant interaction for cell inhibition rates and Annexin V expression levels were observed.After 72 h incubation,the CD11b expression levels (89.46±9.13) ％ in NB4 cells treated with 15 μ mol/L SV in combination with 0.5 μ mol/L ATRA were significantly higher than those treated with ATRA (71.27±7.27) ％ and SV (62.41±6.37) ％ (t =2.71,P =0.054; t =4.37,P =0.017)' solely.Conclusion Simvastatin in vitro inhibits NB4 cell proliferation,promotes cell apoptosis,and synergistically induces cell differentiation with ATRA dose-dependently in vitro,which indicates that SV may have the effect of synergistic anti-promyelocytic potency with ATRA.

Comparison of morphology, immunology and cytogenetics in adult acute leukemia of different ages / 白血病·淋巴瘤

Objective To investigate the characters of morphology,immunology and cytogenetics of adult acute leukemia (AL) in different ages. Methods 172 cases of newly diagnosed adult AL were divided into two groups:the non-aged group (age＜60 years) and the aged group ( age≥60 years). Morphology,immunology and cytogenetics between the two groups were compared. Results The incidence of M3 in aged AL was significantly lower than that in non-aged AL[6.0 %(3/50) vs 18.9 %(23/122),P ＜0.05]. The incidence of hypo-or extremely hypo-cellular AL in aged AL was significantly higher than that in non-aged AL[14.0 %(7/50) vs 4.1 %(5/122),P ＜0.05],but the incidence of hyper-or extremely hyper-cellular was significantly lower than that in non-aged AL[52.0 %(26/50) vs 73.8 %(90/122),P ＜0.05]. Among aged acute myeloid leukemia (AML),the incidence of lymphoid antigen positive AML (Ly+AML) was significantly higher than that in non-aged AML[63.4 %(26/41) vs 41.3 %(38/92),P ＜0.05]. The incidence of adverse karyotypes in aged AML was significantly higher than that in non-aged AML[33.3 %(11/33) vs 14.1 %(10/71),P ＜0.05].Conclusion Age is an important prognostic factor in AL. Generally,aged AL has poorer prognosis than nonaged AL.

Molecular cytogenetic abnormalities in chronic lymphocytic leukemia detected by interphase fluorescence in situ hybridization / 白血病·淋巴瘤

Objective To investigate molecular cytogenetic abnormalities in chronic lymphocytic leukemia and clinic prognostic significance. Methods Conventional cytogenetics (CC) examination was performed in 17 cases with CLL by I-FISH with five probes [DI3S25(13q14.3), ATM(11q22.3), RB1(13q14), p53(17p13.1) and CSP12(12p11.1-12q11.1)]to detect molecular cytogenetic abnormalities in CLL. Results Among 17 cases of CLL, by CC examination, only 18.75 % patient were found to have chromosomal abnormalities;whereas on I-FISH, 70.6 % patient were found to have molecular cytogenetic abnormalities including 13q-(47.1%) del(RB1) (23.5 %), del(13q13.4)(29.4 %), trisomy 12 (29.4%), del(17p13.1)(11.8 %), del (ATM)(5.6 %), the frequency of complex abnormalities were 11.8 %. No correlation of molecular cytogenetic abnormalities with sex, age, Binet stage, LDH and β_2-MG were found. Conclusion I-FISH is a more rapid, accurate and sensitive technique for detection of molecular cytogenetic abnormalities in CLL than CC, There was no statistically significant difference between molecular cytogenetic abnormalities and clinic characteristics, but its prognostic significance in CLL needs to be further investigated.