IL-10基因单核苷酸多态性与急性淋巴细胞白血病易感性的相关性研究

Objective To observe the relationship of IL-10 gene single nucleotide polymorphism and the susceptibility to ALL. Methods The bone marrow and peripheral blood samples from 115 ALL patients and 323 healthy controls were collected in Peking University First Hospital and Beijing Dao-pei Hospital from January 2007 to December 2009. The DNA were extracted from all samples. The primers of -819C/T and -592A/C in the promoter region of IL-10 gene were designed for the PCR. The restrictive fragment length polymorphism of IL-10 gene was analyzed by using restrictive enzyme Msl Ⅰ and HpyCH4 Ⅲ.Sequencing was done in part of these samples to confirm the results of PCR. The differences of genotypes and allele ratio of -819 and -592 sites were analyzed between the ALL patients and healthy controls. Real-time quantitative PCR was performed to detect the EB virus (EBV) infection and the expression of BCR/ABL fusion gene. The differences of genotypes and allele ratio of -819 and -592 sites were analyzed between the positive and negative group. Results The genotype ratios of -819CC, -819TT, - 819CT, -592AA,- 592CC and - 592AC were 14. 8% ( 17/115 ), 45.2% ( 52/115 ), 40. 0% ( 46/115 ), 43.5% ( 50/115 ),16. 5% ( 19/115 ), 40. 0% ( 46/115 ) in ALL patients, and were 9. 9% ( 32/323 ), 16. 4% ( 53/323 ),73.7% ( 238/323 ), 11.8% ( 38/323 ), 15.5% ( 50/323 ), 72. 8% ( 235/323 ) in the healthy controls,respectively. The genotypes of -819 and -592 sites had statistically significant differences between the two groups(x2 values were 46.000 and 54.550, all P ＜ 0. 05 ). The allele ratio of -819T and -592A were (65.2%, 150/230) and (63.5%, 146/230) in ALL patients, while they were 53.5% (344/646) and 48. 1% (311/646)in the healthy controls. There were statistically significant differences between the two groups (x2 values were 9. 877 and 15.986, all P ＜ 0. 05 ). The EBV DNA were detected in 42 ALL patients,among which 22 were positive and 20 were negative. The genotype ratios of -819CC, -819TT, -819CT,-592AA, - 592CC, - 592AC in EBV positive group were 9. 1% ( 2/22 ), 40. 9% ( 9/22 ), 50. 0%(11/22) ,31.8% ( 7/22 ), 13.6% ( 3/22 ), 54. 5% ( 12/22 ), while they were 35.0% ( 7/20 ), 45.0%(9/20) ,20. 0% (4/20) ,35.0% (7/20) ,45.0% (9/20) ,20. 0% (4/20) in the EBV negative group. The genotypes of -819 and -592 sites showed no statistical differences between the two groups( all P ＞ 0. 05 ).The BCR/ABL fusion gene were detected in 36 ALL patients, among which 20 were positive and 16 were negative. The genotype ratios of - 819CC, - 819TT, - 819CT, - 592AA, - 592CC, - 592AC in BCR/ABL positive group were 0% (0/20) ,45.0% (9/20) ,55.0% ( 11/20), 45. 0% (9/20) ,5.0% (1/20) ,50. 0%( 10/20), while they were 18. 8% ( 3/16 ), 50. 0% ( 8/16), 31.3% ( 5/16 ), 50. 0% ( 8/16 ), 18. 8%(3/16), 31.3 % (5/16)in the BCR/ABL negative group. The genotypes of -819 and -592 sites showed no statistical differences between the two groups ( all P ＞ 0. 05 ). Conclusion The population with - 819TT and - 592AA genotype of IL-10 gene shows susceptibility to ALL.     

IL-10基因单核苷酸多态性与急性淋巴细胞白血病易感性的相关性研究

Objective To observe the relationship of IL-10 gene single nucleotide polymorphism and the susceptibility to ALL. Methods The bone marrow and peripheral blood samples from 115 ALL patients and 323 healthy controls were collected in Peking University First Hospital and Beijing Dao-pei Hospital from January 2007 to December 2009. The DNA were extracted from all samples. The primers of -819C/T and -592A/C in the promoter region of IL-10 gene were designed for the PCR. The restrictive fragment length polymorphism of IL-10 gene was analyzed by using restrictive enzyme Msl Ⅰ and HpyCH4 Ⅲ.Sequencing was done in part of these samples to confirm the results of PCR. The differences of genotypes and allele ratio of -819 and -592 sites were analyzed between the ALL patients and healthy controls. Real-time quantitative PCR was performed to detect the EB virus (EBV) infection and the expression of BCR/ABL fusion gene. The differences of genotypes and allele ratio of -819 and -592 sites were analyzed between the positive and negative group. Results The genotype ratios of -819CC, -819TT, - 819CT, -592AA,- 592CC and - 592AC were 14. 8% ( 17/115 ), 45.2% ( 52/115 ), 40. 0% ( 46/115 ), 43.5% ( 50/115 ),16. 5% ( 19/115 ), 40. 0% ( 46/115 ) in ALL patients, and were 9. 9% ( 32/323 ), 16. 4% ( 53/323 ),73.7% ( 238/323 ), 11.8% ( 38/323 ), 15.5% ( 50/323 ), 72. 8% ( 235/323 ) in the healthy controls,respectively. The genotypes of -819 and -592 sites had statistically significant differences between the two groups(x2 values were 46.000 and 54.550, all P ＜ 0. 05 ). The allele ratio of -819T and -592A were (65.2%, 150/230) and (63.5%, 146/230) in ALL patients, while they were 53.5% (344/646) and 48. 1% (311/646)in the healthy controls. There were statistically significant differences between the two groups (x2 values were 9. 877 and 15.986, all P ＜ 0. 05 ). The EBV DNA were detected in 42 ALL patients,among which 22 were positive and 20 were negative. The genotype ratios of -819CC, -819TT, -819CT,-592AA, - 592CC, - 592AC in EBV positive group were 9. 1% ( 2/22 ), 40. 9% ( 9/22 ), 50. 0%(11/22) ,31.8% ( 7/22 ), 13.6% ( 3/22 ), 54. 5% ( 12/22 ), while they were 35.0% ( 7/20 ), 45.0%(9/20) ,20. 0% (4/20) ,35.0% (7/20) ,45.0% (9/20) ,20. 0% (4/20) in the EBV negative group. The genotypes of -819 and -592 sites showed no statistical differences between the two groups( all P ＞ 0. 05 ).The BCR/ABL fusion gene were detected in 36 ALL patients, among which 20 were positive and 16 were negative. The genotype ratios of - 819CC, - 819TT, - 819CT, - 592AA, - 592CC, - 592AC in BCR/ABL positive group were 0% (0/20) ,45.0% (9/20) ,55.0% ( 11/20), 45. 0% (9/20) ,5.0% (1/20) ,50. 0%( 10/20), while they were 18. 8% ( 3/16 ), 50. 0% ( 8/16), 31.3% ( 5/16 ), 50. 0% ( 8/16 ), 18. 8%(3/16), 31.3 % (5/16)in the BCR/ABL negative group. The genotypes of -819 and -592 sites showed no statistical differences between the two groups ( all P ＞ 0. 05 ). Conclusion The population with - 819TT and - 592AA genotype of IL-10 gene shows susceptibility to ALL.     

IL-10基因位点多态性与ACS的相关性分析

[目的]探讨急性冠脉综合征(ACS)与白介素-10(IL-10)基因位点4259A/G、-1082G/A、-592C/A、-819C/T多态性分布的相关性.[方法]选择2010年5月至2015年9月常州市溧阳人民医院心内科和武进人民医院心内科收治的ACS患者471例(观察组),选择同期两个医院心内科收治的197例非ACS者为对照组,采用基因测序技术检测所有标本IL-10基因型,分析其与ACS发病相关危险因素.[结果]两组在年龄、吸烟、合并糖尿病、血清TC、TG、LDL-C方面比较,差异均具有统计学意义(P<0.05).在两组研究对象中,IL-104259A/G位点仅发现AA一种基因型;但是在两组研究对象中发现IL-10-1082G/A位点上有GG、AG、AA三种基因型、IL-10-582C/A位点上有CC、AC和AA三种基因型、IL-10-819C/T位点上有CC、CT和TT三种基因型.与对照组作比较后,尚未发现各种基因型之间分布频率、等位基因分布频率有统计学差异;对每一种基因型行引起ACS发病传统危险因素多元Logistic回归分析显示IL-10基因-592C/A、-819C/T、-1082G/A基因型与ACS的发病无相关性.[结论]在常州地区心血管疾病住院患者IL-10基因4259A/G仅有AA一种基因型;IL-10基因-1082G/A、-592C/A、-819C/T具有多态性,但其多态性分布与本地区汉族人群ACS的发病无相关性.     

肿瘤坏死因子-α-863位点基因多态性与痛风相关性的初步研究

目的 探讨肿瘤坏死因子-α(TNF-α)-863位点基因多态性与唐山地区汉族人群痛风的相关性.方法 以聚合酶链反应-连接酶检测反应(PCR-LDR)方法对80例痛风患者(病例组)和80例正常对照者(对照组)进行TNF-α-863位点的单核苷酸多态性分型,比较各基因型及等位基因频率在正常人群和痛风患者中的差异.结果 2组之间TNF-α-863位点各基因型分布频率,病例组与对照组A/A基因型频率相同(均为3.8%),病例组与对照组C/A基因型频率分别为23.8%与38.8%,C/C基因型频率分别为72.5%与57.5%,2组各基因型频率差异无统计学意义(x2=4.2646,P=0.1187);病例组与对照组C等位基因频率分别为84.4%与76.9%,差异无统计学意义(x2=2.8807,P=0.0897).结论 TNF-α-863位点基因多态性可能与唐山汉族人群痛风易感性无关.

Abstract：

Objective To investigate the association between the tumour necrosis factor-α-863(TNF-α-863) polymorphism and gout in Han population from the city of Tangshan.Methods We recruited 80 gout patients and 80 healthy individuals into this study.The polymorphisms of TNF-α-863 site were analyzed by polymerase chain reaction-ligase detection reaction(PCR-LDR).The frequencies of different TNF-α-863 genotypes/alleles were analyzed in the gout group and the control subjects.Results No significant differences were observed in the genotype frequencies(x2=2.8807,P=0.0897) and allelic frequencies(x2=4.2646,P=0.1187) of TNF-α-863 site in the comparison between gout and control groups.Conclusion The result of our study suggests that the polymorphism of TNF-α-863 site may not related to gout in Han population in Tangshan.     

酶切富集联合DHPLC检测肿瘤患者外周血EGFR和KRAS基因突变及其临床应用

Objective To establish a REDE-DHPLC method for detecting the EGFR and KRAS mutations in plasma DNA from tumor patients, and investigate its clinical significance. Methods Restriction endonucleases Mse Ⅰ , Msc Ⅰ , BstN Ⅰ and Bgl Ⅰ were used to digest the wild type fragments of exon 19,exon 21 of EGFR gene and coden 12, 13 of KRAS gene for enriching the mutation fragments, and REDE-DHPLC method was established to detect EGFR and KRAS mutations. The sensitivities of REDE-DHPLC and conventional DHPLC were analyzed by using a series of plasmids containing 50%, 10%, 5%, 1% and 0. 1% mutation genes. Then, Plasma samples and paraffin-embedded tissue samples of 120 NSCLC patients and 120 colorectal cancer patients were detected by REDE-DHPLC. Compared with conventional DHPLC and sequencing, the diagnostic efficiency of REDE-DHPLC method was evaluated by detecting the mutation status of 2 genes in plasma of NSCLC and colorectal cancer patients. Results The sensitivity values of REDE-DHPLC and conventional DHPLC for detecting mutations in 4 loci were 0. 1% and 1%respectively. Plasmid DNA containing 0.1% mutation gene was detected to be positive continually for 2 to 3 times by REDE-DHPLC. EGFR mutation rates of 120 plasma from NSCLC patients detected by REDE-DHPLC, conventional DHPLC and sequencing methods were 27. 5%, 16. 7% and 12.5% respectively, and KRAS mutation rates of 120 plasma from colorectal cancer patients were 38. 3%, 25. 8% and 16. 7%,respectively. The positive rates of EGFR and KRAS mutation detected by REDE-DHPLC were significantly higher than conventional DHPLC(x2 = 4. 092, 4. 301, all P ＜ 0. 05 ) and sequencing method (x2= 8. 438,14. 127,all P ＜ 0. 05 ). In comparison with conventional DHPLC, the sensitivities of REDE-DHPLC for detecting EGFR and KRAS mutation were 100% (20/20,31/31), the specificities were 87. 0% (87/100)and 83. 2% (74/89). In comparison with sequencing method, the sensitivities of REDE-DHPLC were 100%( 15/15,20/20), the specificities were 82.9% (87/105)and 74. 0% (74/100). The coincidence rate of the two methods for detecting EGFR and KRAS mutation were 89. 2% ( 107/120, Kappa = 0. 690, P ＜ 0. 05 ) and 87.5% ( 105/120, Kappa= 0. 718, P ＜ 0. 05 ). The Consistency of EGFR and KRAS mutation status in plasma and tissues detected by REDE-DHPLC were 91.7% (33/36, Kappa =0. 939,P ＜0. 05)and 90. 2 %(46/51, Kappa = 0. 914, P ＜ 0. 05 ), respectively. Conclusions The REDE-DHPLC method is highly sensitive and specific for detecting EGFR and KRAS mutations in plasma DNA from tumor patients. The results are easy to be interpreted without missing homozygous point mutation, which indicate that the detection of EGFR and KRAS mutations in plasma DNA by REDE-DHPLC could therefore extend to be usedin clinical laboratory.     

酶切富集联合DHPLC检测肿瘤患者外周血EGFR和KRAS基因突变及其临床应用

Objective To establish a REDE-DHPLC method for detecting the EGFR and KRAS mutations in plasma DNA from tumor patients, and investigate its clinical significance. Methods Restriction endonucleases Mse Ⅰ , Msc Ⅰ , BstN Ⅰ and Bgl Ⅰ were used to digest the wild type fragments of exon 19,exon 21 of EGFR gene and coden 12, 13 of KRAS gene for enriching the mutation fragments, and REDE-DHPLC method was established to detect EGFR and KRAS mutations. The sensitivities of REDE-DHPLC and conventional DHPLC were analyzed by using a series of plasmids containing 50%, 10%, 5%, 1% and 0. 1% mutation genes. Then, Plasma samples and paraffin-embedded tissue samples of 120 NSCLC patients and 120 colorectal cancer patients were detected by REDE-DHPLC. Compared with conventional DHPLC and sequencing, the diagnostic efficiency of REDE-DHPLC method was evaluated by detecting the mutation status of 2 genes in plasma of NSCLC and colorectal cancer patients. Results The sensitivity values of REDE-DHPLC and conventional DHPLC for detecting mutations in 4 loci were 0. 1% and 1%respectively. Plasmid DNA containing 0.1% mutation gene was detected to be positive continually for 2 to 3 times by REDE-DHPLC. EGFR mutation rates of 120 plasma from NSCLC patients detected by REDE-DHPLC, conventional DHPLC and sequencing methods were 27. 5%, 16. 7% and 12.5% respectively, and KRAS mutation rates of 120 plasma from colorectal cancer patients were 38. 3%, 25. 8% and 16. 7%,respectively. The positive rates of EGFR and KRAS mutation detected by REDE-DHPLC were significantly higher than conventional DHPLC(x2 = 4. 092, 4. 301, all P ＜ 0. 05 ) and sequencing method (x2= 8. 438,14. 127,all P ＜ 0. 05 ). In comparison with conventional DHPLC, the sensitivities of REDE-DHPLC for detecting EGFR and KRAS mutation were 100% (20/20,31/31), the specificities were 87. 0% (87/100)and 83. 2% (74/89). In comparison with sequencing method, the sensitivities of REDE-DHPLC were 100%( 15/15,20/20), the specificities were 82.9% (87/105)and 74. 0% (74/100). The coincidence rate of the two methods for detecting EGFR and KRAS mutation were 89. 2% ( 107/120, Kappa = 0. 690, P ＜ 0. 05 ) and 87.5% ( 105/120, Kappa= 0. 718, P ＜ 0. 05 ). The Consistency of EGFR and KRAS mutation status in plasma and tissues detected by REDE-DHPLC were 91.7% (33/36, Kappa =0. 939,P ＜0. 05)and 90. 2 %(46/51, Kappa = 0. 914, P ＜ 0. 05 ), respectively. Conclusions The REDE-DHPLC method is highly sensitive and specific for detecting EGFR and KRAS mutations in plasma DNA from tumor patients. The results are easy to be interpreted without missing homozygous point mutation, which indicate that the detection of EGFR and KRAS mutations in plasma DNA by REDE-DHPLC could therefore extend to be usedin clinical laboratory.     

肺纤维化大鼠血清克拉拉细胞蛋白和表面活性蛋白-D动态变化及早期诊断意义

Objective To detect the dynamic changes of the level of serum Clara cell protein(CC16)and surfactant protein-D(SP-D)in rats with pulmonary fibrosis induced by bleomycin and to evaluate their value in early diagnosis of pulmonary fibrosis. Methods Sixty Wistar rats were randomly divided into two groups, the control group and bleocin-induced pulmonary fibrotic group,with 30 rats in each group. The rats were killed at 1,3,7,14 and 28 days of treatment Pathology changes of lung tissue were observed by HE,Masson stain,alkaline hydrolysis to assess the hydroxyproline concentration of lung tissue, enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of serum CC16 and SP-D. Results The hydroxyproline concentration of lung tissue in the pulmonary fibrotic group ((913. 1 ±69. 3) μg/g) were higher than those of the control group ((790. 5 ± 36. 8) μg/g) from the seventh day(P <0. 05). The levels of serum CC16 of the pulmonary fibrotic group((27. 34 ± 0. 32) μg/L) were lower than those of the control group((27. 85 ±0. 32)μg/L) since the third day(P<0. 05) ,and tended to decrease with the development of the disease. However,the levels of SP-D of the former group were always higher(P <0. 05), and tended to increase with the development of the disease. Conclusions The levels of serum CC16 and SP-D changed considerably in early-stage of pulmonary fibrosis, thus might be used as biomarker for early diagnosis and have some value for pathogenesis of pulmonary fibrosis.     

肺纤维化大鼠血清克拉拉细胞蛋白和表面活性蛋白-D动态变化及早期诊断意义

Objective To detect the dynamic changes of the level of serum Clara cell protein(CC16)and surfactant protein-D(SP-D)in rats with pulmonary fibrosis induced by bleomycin and to evaluate their value in early diagnosis of pulmonary fibrosis. Methods Sixty Wistar rats were randomly divided into two groups, the control group and bleocin-induced pulmonary fibrotic group,with 30 rats in each group. The rats were killed at 1,3,7,14 and 28 days of treatment Pathology changes of lung tissue were observed by HE,Masson stain,alkaline hydrolysis to assess the hydroxyproline concentration of lung tissue, enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of serum CC16 and SP-D. Results The hydroxyproline concentration of lung tissue in the pulmonary fibrotic group ((913. 1 ±69. 3) μg/g) were higher than those of the control group ((790. 5 ± 36. 8) μg/g) from the seventh day(P <0. 05). The levels of serum CC16 of the pulmonary fibrotic group((27. 34 ± 0. 32) μg/L) were lower than those of the control group((27. 85 ±0. 32)μg/L) since the third day(P<0. 05) ,and tended to decrease with the development of the disease. However,the levels of SP-D of the former group were always higher(P <0. 05), and tended to increase with the development of the disease. Conclusions The levels of serum CC16 and SP-D changed considerably in early-stage of pulmonary fibrosis, thus might be used as biomarker for early diagnosis and have some value for pathogenesis of pulmonary fibrosis.     

IKZF3基因单核苷酸多态性与儿童急性淋巴细胞白血病的关系

目的:探讨IKAROS家族锌指蛋白3(IKZF3)基因单核苷酸多态性与儿童急性淋巴细胞白血病(ALL)发病风险的关系。方法:收集286例ALL患儿以及382例健康儿童的外周血样本,分别作为ALL组和对照组。采用Taq Man探针实时定量PCR技术检测IKZF3基因rs62066988 C T和rs12946510 C T两个单核苷酸多态性位点的基因型分布情况,并分析二者与ALL的相关性。结果:rs62066988位点的CC、CT和TT基因型在ALL组的分布频率分别为58.39%、37.06%和4.55%,在对照组的分布频率分别为69.19%、27.68%和3.13%。rs12946510位点的基因型CC、CT和TT在ALL组的分布频率分别为58.16%、34.75%和7.09%,在对照组的分布频率分别为55.76%、37.43%和6.81%。与对照组相比,rs62066988位点的CT/TT基因型在ALL组中的分布频率显著增加(CT/TT vs CC:校正后OR=1.59,95%CI:1.16-2.19,P=0.004),而rs12946510 C T的多态性分布在ALL组和对照组之间并无显著差异。结论:IKZF3基因rs62066988位点的CT/TT基因型与儿童ALL发病风险的增加有关。     

Association between matrix metalloproteinase-9 polymorphism (-1562C ＞ T/R279Q) and acute coronary syndrome in Uygur nationality of Xinjiang Autonomous Region of China

Objective To investigate the association between matrix metalloproteinase-9 (MMP-9) gene polymorphism (-1562C ＞ T/R279Q) and acute coronary syndrome (ACS) in Uygur nationality of Xinjiang Autonomous Region of China. Methods A total of 352 patients with ACS including 213 patients with unstable angina pectoris and 139 patients with acute myocardial infarction evidenced by using coronary arteriography and 421 control subjects were recruited in this study. The MMP-9-1562C ＞ T and R279Q genotypes were detemined by using PCR-RFLP method. The relationship between the polymorphism in the MMP-9 gene and the severity of coronary arterial stenosis was analyzed. All polymorphisms were determined for confimation with Hardy-Weinberg expectations in both groups separately. Differences in distributions of genotypes and alleles between two groups were analyzed with x2 test. The association between the MMP-9 polymorphisms and the risk of ACS was estimated by odds ratio(Ors) and their 95% confidence intervals (CIs), and the comprehensive evaluation of the factors associated with ACS was determined by using multifactor logistic regression. P ＜ 0. 05 was considered to be statistically significant. Results The genotype frequencies for CT + TT genotypes and T allele were 25.9 and14.5 percent in ACS subjects and 15.7 and 8.4 percent in control subjects, respectively. The genotype frequencies were different significantly between the two groups (x2 = 12.26,P ＜ 0.01;x2 = 14.15,P ＜ 0.01, respectively). No relationship between R279Q polymorphism and ACS was found in this study ( P ＞ 0.05). The multifactor logistic regression analysis showed that the T allele carrier (CT + TT) significantly increased the risk of ACS compared with the CC genotype ( OR = 1.791,95 % CI: 1. 088 - 2.951, P = 0.022) after adjustment for tradition risk factors. The frequencies for CT + TT and CC genotypes of the -1562C ＞ T polymorphism were not statistically different among ACS patients with one, two and three or more significantly diseased vessels ( x2 = 1.15, P = 0.56). Conclusions The findings suggest that the polymorphism in MMP-9 gene promoter (-1562C ＞ T) is associated with the susceptibility to the ACS. The T allele might be an independent risk factor for the ACS. But the -1562C ＞ T polymorphism may not be useful as a predictor of the severity of coronary arterial stenosis. The R279Q polymorphism of MMP-9 gene was not significantly associated with ACS in this studied population.     

骨髓形态学染色联合荧光原位杂交技术在急性白血病鉴别诊断中的应用

目的 探讨联合骨髓形态学染色和荧光原位杂交(MGG-FISH)技术在Ph染色体阳性急性淋巴细胞白血病(Ph+ALL)和慢性粒细胞白血病急性淋巴细胞白血病变(CML-LBC)鉴别诊断中的临床应用价值.方法 应用BCR(绿色)和ABL(橘红色)双色双融合探针,通过MGG-FISH技术对4例Ph+ALL患者、4例CML-LBC患者、1例CML急性白血病变合并淋巴瘤、1例CML急性混合细胞白血病变患者的骨髓涂片标本进行检测,依据形态学分别检测10份标本红细胞系、粒细胞系和淋巴细胞系的BCR/ABL融合信号阳性细胞百分率.结果 通过MGG-FISH方法分析,4份Ph+ALL骨髓标本红细胞系未累及,BCR/ABL融合信号阳性细胞百分率均为0%;粒细胞系阳性率分别为11%(1/9)、8%(1/12)、0%(0/8)、10%(1/10);淋巴细胞系阳性率分别为97%(76/78)、98%(87/89)、98%(97/99)、97%(75/77).4份CML-LBC骨髓标本红细胞系阳性率分别为100%(8/8)、91%(10/11)、82%(9/11)、88%(7/8);粒细胞系阳性率分别为89%(8/9)、96%(94/98)、100%(47/47)、98%(40/41);淋巴细胞系阳性率分别为96%(78/81)、93%(52/56)、96%(68/71)、95%(58/61).其余2份标本阳性率均超过80%,且通过MGG-FISH分析鉴定了恶性克隆的来源.结论 MGG-FISH技术可以准确、快速地对原发性Ph+ALL和CML-LBC进行鉴别,并可进行克隆源性分析.     

CD25在急性B淋巴细胞白血病中的表达及其临床意义探讨

本研究旨在通过流式细胞术标记CD25,探讨其与急性B淋巴细胞白血病（B—ALL）的关系及其临床意义。选择88例初发B—ALL患者骨髓,应用流式细胞术检测免疫表型标记,包括白介素2受体α链（CD25）、B链（CD122）、Y链（CD132）,CD19、CD20、CD10、CD34、CDIgM、CD79a、CD22、CDTDT;用定性PCR方法检测BCR／ABL融合基因表达,实时定量PCR检测IL2RA（CD25基因）的表达。结果表明,CD25＋和CD25-的B—ALL患者白细胞计数（WBC）、血红蛋白（Hb）、血小板（Plt）、骨髓原始细胞比例（marrowblast％）、外周血原始细胞比例（peripheralblast％）、肝脾和淋巴结肿大、cD10、cD20、cD22、cD34、CD79a、CDTDT、CDIgM表达水平差别均无统计学意义（P〉0．05）,而CD19在CD25＋组中表达水平高于CD25-组（P〈0．05）。本组BCR／ABL＋患者为21例（23．9％,21／88）,CD25＋表达率为66．7％（14／21）;BCR／ABL-患者为67例,CD25＋表达率为4．5％（3／67）,两组间差别有统计学意义（P〈0．05）。IL2RA基因mRNA表达水平在CD25＋和CD25-两组间无统计学差异（P〉0．05）。21例BCR／ABL＋B—ALL患者中,CD25＋与CD25-两组间在缓解率和复发率方面差异无统计学意义（P〉0．05）。BCR／ABL＋B—ALL患者中有8例复发,复发率为38．1％（8／21）,BCR／ABL。组有9例复发,复发率为13．4％（9／67）,两组间差异有统计学意义（P〈0．05）。BCR／ABL＋组无复发生存时间（relapse—freesurvival,RFS）为21个月,BCR／ABL＋CD25＋患者RFS时间为15个月,而BCR／ABL＋CD25-患者RFS时间为21个月,BCR／ABL—CD25-患者RFS时间为24个月。结论：CD25在BCR／ABL＋B-ALL上患者中呈高水平表达,可以作为B—ALL的BCR／ABL融合基因表达的预测指标,与疾病复发有关,CD25＋可以作为判断B—ALL不良预后的辅助标志。     

BCR/ABL探针在骨髓增殖性疾病中的临床运用

本研究通过回顾分析荧光原位杂交(FISH)检测BCR/ABL融合基因结果,探讨其在慢性骨髓增殖性疾病(CMPD)和Ph+急性淋巴细胞白血病(Ph+ALL)的鉴别诊断及治疗后微小残留病(MRD)动态监测中的运用价值。初诊和治疗后病例分别使用BCR/ABL(ES)和BCR/ABL(DF)探针检测BCR/ABL融合基因。结果表明:初诊CMPD 49例,形态学符合CML骨髓象28例,确诊CML 23例,形态符合率23/28(82.1%),BCR/ABL阳性23/23,敏感度、特异度均为100%。确诊病例BCR/ABL阳性细胞率为81.3%±17.7%;allo-HSCT 13例,9例长期无病生存,4例复发,经供者淋巴细胞输注(DLI)、伊马替尼或allo-HSCT治疗后多次监测BCR/ABL阴性;伊马替尼治疗16例,其中11例于1年后多次监测BCR/ABL阴性,5例分别于6、7、10年后监测BCR/ABL阳性,其中1例经allo-HSCT成功,BCR/ABL转阴。结论:FISH技术是一项敏感、特异的诊断技术,针对初诊和治疗后病例分别运用两种不同的探针检测BCR/ABL融合基因,有助于准确、快速鉴别诊断CML、Ph+ALL,动态监测酪氨酸激酶抑制剂治疗和allo-HSCT后MRD。     

Chronic myeloid leukemia: current perspectives

Chronic myeloid leukemia (CML), characterized by the t(9;22) and BCR/ABL1 fusion, is a disease model for studying the mechanisms of genetic abnormalities in leukemogenesis. The detection of the t(9;22), characterization of the BCR/ABL fusion, and the discovery of imatinib have elegantly reflected the success of our research efforts in CML. However, genomic instabilities that lead to the formation of the BCR/ ABL1 fusion are not fully understood. It is important to understand how various genes that are involved in regulating the signaling pathway and epigenetic deregulation cooperate with the BCR/ABL1 fusion in the initiation and progression of CML.     

FTY720, a new alternative for treating blast crisis chronic myelogenous leukemia and Philadelphia chromosome-positive acute lymphocytic leukemia

Blast crisis chronic myelogenous leukemia (CML-BC) and Philadelphia chromosome-positive (Ph1-positive) acute lymphocytic leukemia (ALL) are 2 fatal BCR/ABL-driven leukemias against which Abl kinase inhibitors fail to induce a long-term response. We recently reported that functional loss of protein phosphatase 2A (PP2A) activity is important for CML blastic transformation. We assessed the therapeutic potential of the PP2A activator FTY720 (2-amino-2-[2-(4-octylphenyl)ethyl]-1,3-propanediol hydrochloride), an immunomodulator in Phase III trials for patients with multiple sclerosis or undergoing organ transplantation, in CML-BC and Ph1 ALL patient cells and in in vitro and in vivo models of these BCR/ABL+ leukemias. Our data indicate that FTY720 induces apoptosis and impairs clonogenicity of imatinib/dasatinib-sensitive and -resistant p210/p190(BCR/ABL) myeloid and lymphoid cell lines and CML-BC(CD34+) and Ph1 ALL(CD34+/CD19+) progenitors but not of normal CD34+ and CD34+/CD19+ bone marrow cells. Furthermore, pharmacologic doses of FTY720 remarkably suppress in vivo p210/p190(BCR/ABL)-driven [including p210/p190(BCR/ABL)(T315I)] leukemogenesis without exerting any toxicity. Altogether, these results highlight the therapeutic relevance of rescuing PP2A tumor suppressor activity in Ph1 leukemias and strongly support the introduction of the PP2A activator FTY720 in the treatment of CML-BC and Ph1 ALL patients.     

Differential diagnosis of chronic myeloid leukemia and the related disorders]

Chronic myeloid leukemia(CML) is a generic term that includes five subtypes; i.e. chronic granulocytic leukemia(CGL) (95% of all CML, 90% are Ph+, 5% are Ph-, BCR/ABL+), atypical CML(survival is worse than that of CGL), chronic myelomonocytic leukemia(a subtype of myelodysplastic syndrome), chronic neutrophilic leukemia (Ph-, BCR/ABL-) and juvenile CML(Ph-, BCR/ABL-). It is not so easy to make a diagnosis of Ph-negative CML. Also, about 25% of adult acute lymphoid leukemia(ALL) patients and some essential thrombocythemia patients have Ph chromosome. In addition, about a half of cases with Ph-positive ALL have the same size of BCR/ABL fusion protein as that in Ph-positive CML. It is necessary to distinguish them by the distinctive morphological, cytogenetical and immunological characteristics of these diseases.     

达沙替尼治疗伊马替尼耐药的BCR/ABL阳性白血病的临床研究

本研究评价达沙替尼治疗原发或继发伊马替尼耐药的BCR/ABL阳性白血病的疗效和安全性。对27例原发或继发伊马替尼耐药的慢性髓系白血病(CML)或Ph阳性急性淋巴细胞白血病(Ph+ALL)患者,给予达沙替尼100-140 mg/d口服治疗,评估疗效、总体生存和耐受情况。结果表明:27例伊马替尼耐药的BCR/ABL阳性白血病中位达沙替尼治疗时间8(1-66)个月,中位随访时间54(3-75)个月。27例接受达沙替尼治疗的患者中,88.8%获得完全血液学反应(CHR),44.4%获得主要细胞遗传学反应(mCyR),37%获得完全遗传学反应(CCyR),18.5%获得主要分子学反应(MMR)。伊马替尼耐药的进展期(CML-AP、CML-BC、骨髓复发Ph+ALL)患者接受达沙替尼治疗获CCyR率低于疾病稳定期(CML-CP、骨髓缓解Ph+ALL)患者(P=0.0377),且3-4级不良反应发生率明显增高。达沙替尼治疗后获得CCyR的患者生存期(OS)较未达CCyR者明显延长(63个月vs 9个月,P=0.0126)。达沙替尼治疗后最常见的3-4级不良反应包括血液学反应如血小板减少(51.8%)、中性粒细胞减少(48.1%)、贫血(33.3%)和非血液学反应如胸腔积液(18.5%)、肺部感染(18.5%)、心包积液(11.1%)。3-4级不良反应主要发生在疾病进展期时改服达沙替尼者,均发生于服药12个月内。结论:达沙替尼治疗伊马替尼耐药的BCR/ABL阳性白血病有效,且在疾病稳定期改服达沙替尼疗效和耐受性更好。