急性白血病患者WT1基因定量监测与预后的关系

目的:通过检测WT1基因在成人急性髓系白血病(非APL)中的表达,探讨其在中、高危急性髓系白血病治疗反应和预后的临床应用价值。方法:选取急性髓系白血病患者63例,其中不包含急性早幼粒细胞白血病患者;应用SYBR Green I定量RT-PCR方法检测63例初治急性髓系白血病患者中WT1基因的表达。结果:WT1基因在急性髓系白血病中高表达,在低危组中的表达明显低于中危组和高危组(P 0.05)。WT1基因在中危组与高危组中的表达无明显统计学差异;中、高危组病人经1疗程正规治疗后,缓解组的WT1基因表达量与未缓解组相比明显下降(P 0.05)。在中、高危组病人中,首次治疗未缓解的病人,给予再次诱导治疗达到缓解病人的WT1基因表达量明显低于未缓解组(P 0.05)。在中高危组病人中,WT1低表达的患者治疗后2年总体生存时间(OS)较未缓解病人明显延长(P 0. 05)。结论:检测WT1基因的表达可有助于急性髓系白血病患者的低、中、高危分群,也有助于评价中、高危急性髓系白血病的治疗及预后情况。     

肾母细胞瘤1基因联合多参数流式细胞术检测微小残留病灶评估急性髓系白血病患儿预后的价值

目的 分析肾母细胞瘤1(WT1)基因联合多参数流式细胞术检测微小残留病灶(FCM-MRD)对急性髓系白血病(AML)患儿预后的预测价值。方法 回顾性分析76例AML患儿的临床资料及一般信息。患儿治疗前均采用实时荧光定量聚合酶链式反应(qRT-PCR)检测WT1基因表达,并经FCM检测MRD。所有患儿随访1年,根据预后情况的不同分为预后良好组(n=40)和预后不良组(n=36)。观察2组患儿治疗前及治疗后3、9、12个月的WT1基因及MRD变化情况;比较不同治疗方案患儿治疗前后WT1基因及MRD变化情况;分析AML患儿临床病理特征与WT1基因表达及FCM-MRD阳性率的关系。采用Spearman相关系数分析WT1基因表达、FCM-MRD阳性率与AML患儿预后的关系;绘制Kaplan-Meier生存曲线,分析WT1基因表达、FCM-MRD阳性率对AML患儿复发的影响及相关性;绘制受试者工作特征(ROC)曲线分析WT1基因、FCM-MRD单一及联合检测对AML患儿预后的预测效能;分析WT1基因、MRD与AML患儿FLT3 ITD/TKD突变的关系。结果 预后良好组患儿治疗后9、12个月的WT...     

实时定量RT-PCR检测急性白血病患者骨髓细胞WT1基因表达

目的探讨急性白血病 (AL)患者骨髓细胞WT1基因表达水平。方法建立实时定量RT PCR方法 ,检测了 10 8例AL患者和 2 3例非白血病患者骨髓细胞WT1基因及内参照 β 胆色原脱氢酶 (GAPDH)基因的表达水平。结果初诊与复发AL患者骨髓细胞WT1基因中位表达水平经GAPDH校正后分别为 75 .10和 89.5 6 ,明显高于完全缓解组和对照组 (分别为 2 .0 7和 1.5 1) ,对照组与完全缓解组之间及初诊与复发组之间无统计学差异 ;70例初诊AL中急性淋巴细胞白血病、M1、M2 、M3 亚型WT1基因表达水平明显高于M5,即粒系表达明显高于单核系 ,且WT1基因表达水平与白细胞计数、骨髓原始细胞比例、mdr1基因表达无相关性 ,但与染色体核型有关。对其中 2例患者动态检测WT1基因表达可提示白血病复发情况。结论WT1基因在AL患者骨髓细胞高表达 ,可作为白血病疗效评价及监测残留病灶的指标。     

急性白血病患者WT1基因表达及其临床意义

目的 观察WT1基因在白血病细胞的表达及其临床意义,探讨WT1基因表达与白血病疗效关系。方法 用逆转录-聚合酶链反应（RT—PCR）检测初治及复发急性白血病患者73例治疗前WT1基因的表达,结合临床观察WT1表达阳性与急性白血病治疗疗效关系,并观察缓解后患者WT1基因表达的变化。结果 73例急性白血病患者WT1阳性53例（72．6％）,其中46例急性髓细胞白血病阳性32例（69．6％）,23例急性淋巴细胞白血病阳性17例（73．9％）。3例急性混合细胞白血病及1例急性浆细胞白血病均表达阳性。在急性淋巴细胞白血病和非M3型急性髓细胞白血病中,WT1表达阳性对治疗缓解率无明显影响。对21例患者治疗后动态观察,当完全缓解时,WT1表达转阴性;复发时WT1表达又转阳性。结论 WT1在急性白血病阳性率较高,但阳性与否对临床疗效无明显影响。WT1基因作为白血病标志用于急性白血病微小残留病的检测有重要价值。     

白血病患者WT1基因表达及其临床意义观察

目的：观察WT1基因在白血病细胞中的表达及其临床意义，初步探讨WT1基因表达与白血病疗效关系。方法：采用逆转录～聚合酶链法(RT-PCR)检测治疗前47例各类型和白血病患者WT1基因的表达，并观察缓解后患者WT1基因表达的变化，并对33例WT1表达阳性者进行单链构象多态性分析(SSCP)分析。结果：急性白血病37例中30例(81．08％)WT1表达阳性，其中急性髓细胞白血病16／19例(84．21％)阳性，急性淋巴细胞白血病14／18例(77．78％)阳性；慢性粒细胞白血病(CML)急变期3／5例阳性，CML慢性期5例均阴性。WT1表达阳性对治疗缓解率无明显影响。对WT1表达阳性的21例急性白血病治疗后WT1表达动态观察，当获得完全缓解时，WT1表达转阴性；在白血病复发前及复发时，WT1表达转阳性。SSCP分析33例WT1阳性表达者第7～10外显子区PCR产物，未发现泳动异位条带。结论：WT1在急性白血病中表达阳性率较高；慢性粒细胞白血病慢性期未见WT1表达，急变期阳性率增高；WT1表达阳性对治疗完全缓解无明显影响。WT1基因作为泛白血病标志用于白血病微小残留病灶的检测有重要实用价值。     

荧光定量PCR检测急性白血病患者外周血 WT1基因表达及其临床意义

为研究WT1基因表达与临床疗效和预后的关系,探讨其在白血病微小残留病检测中的作用,用荧光定量RT-PCR方法检测55例初发白血病患者外周血及10例正常人外周血的WT1基因表达,跟踪20例急性白血病患者外周血WT1基因表达。结果显示,白血病初治组(40例ANLL、15例ALL)与正常人外周血WT1基因表达有显著差异(P<0.001)。在急性白血病患者中,WT1≤6.8×10-3组生存期长于WT1>6.8×10-3组(P=0.027);白血病患者初发时WT1呈高度表达,完全缓解后,迅速或缓慢下降至少1个对数级,复发时再次增高。跟踪检测20例急性白血病患者外周血WT1基因表达,结果7例复发,5例在临床复发前2-3月WT1的水平明显增高,至少上升0.8个对数级。结论:荧光定量RT-PCR方法检测白血病外周血WT1表达具有简便易行、准确性高、特异性好的特点。与正常人外周血比较,WT1在各类白血病外周血中呈高度表达,且表达水平与预后负相关;对外周血WT1基因表达进行定量分析,可用于微小残留病的监测。     

WT1基因在骨髓增生异常综合征及急性白血病患者中的表达

为了研究WT1基因在骨髓增生异常综合征 (MDS)和急性白血病 (AL)患者中的表达及其与临床预后的关系 ,采用逆转录 聚合酶链反应 (RT PCR)方法检测 2 2例MDS和 6 9例AL的WT1mRNA的表达。结果表明 ,WT1mRNA在MDS RA及MDS RAS中低表达 ,阳性率为 10 % (1/10 ) ;在MDS RAEB和MDS RAEB t中高表达 ,阳性率为 91.7% (11/12 ) ,两者差异有统计学意义 (P <0 .0 1)。WT1mRNA在各型急性白血病中均有表达 ,初发和复发急性白血病中WT1mRNA的阳性率 (6 9% )高于缓解患者 (12 .5 % ) (P <0 .0 1)。初发与复发AL的WT1mRNA的相对表达量无差异 ,而MDS RAEB和RAEB t的表达量低于初发AL患者。WT1mRNA阳性的AML患者的化疗缓解率 (4 1% )低于阴性患者 (78% ) ;WT1基因相对表达量≥ 1的AML患者的CR率 (18% )低于 <1的患者 (5 5 % )。结论 :WT1基因在MDS RAEB和MDS RAEB t中的表达明显高于MDS RA和MDS RAS ,动态监测WT1基因的表达可能有助于监测MDS的病情变化。WT1基因表达与否及表达高低与初发AL的预后有关 ;WT1基因是急性髓细胞白血病的一个独立的预后因素。     

MDR1、SOX4、WT1在急性髓系白血病中的作用及表达水平

目的 探讨多药耐药基因(MDR1)、SOX4、Wilms瘤基因1(WT1)与急性髓系白血病(AML)临床特征及预后的关系。方法 选取2020年1月至2021年8月该院收治的149例AML患者作为AML组,72例轻度贫血患者作为对照组,采用实时荧光定量聚合酶链反应检测骨髓组织中MDR1 mRNA、SOX4 mRNA、WT1 mRNA表达水平。分析MDR1 mRNA、SOX4 mRNA、WT1 mRNA表达水平与AML临床特征及预后的关系。结果 AML组骨髓组织中MDR1 mRNA、SOX4 mRNA、WT1 mRNA表达水平均高于对照组,差异均有统计学意义(P<0.05)。白细胞计数≥40×10⁹/L、有器官浸润、治疗后非完全缓解AML患者骨髓组织中MDR1 mRNA、SOX4 mRNA、WT1 mRNA表达水平均高于白细胞计数<40×10⁹/L、无器官浸润及治疗后完全缓解AML患者,差异均有统计学意义(P<0.05)。MDR1 mRNA、SOX4 mRNA及WT1 mRNA高表达AML患者总生存率均低于MDR1 mRNA、S...     

儿童白血病WT1 MRP基因表达及临床意义

目的　探讨WT1基因、多药耐药相关蛋白基因 (MRP)表达与儿童急性白血病的关系及意义。方法　应用逆转录—聚合酶链反应 (RT -PCR)法测定 4 8例不同类型儿童急性白血病及 10例正常儿童WT1、MRP基因的表达。结果　 4 8例儿童急性白血病患者中 15例WT1基因表达阳性。WT1基因表达阳性患者的完全缓解率 (CR)低于表达阴性者 (分别为 4 4 %和81 8% ,P <0 0 5 )。 10例正常儿童WT1基因表达阴性。MRP基因表达阳性者 18例 ,MRP基因表达阳性者CR低于表达阴性者 (分别为 33 3%和 80 % ,P <0 0 5 )。 10例正常对照基因表达阴性。WT1基因表达阳性患者的MRP阳性率与WT1基因表达阴性者无明显差异 (分别为 5 5 %和 4 2 86 % ,P >0 0 5 )。统计学提示 :WT1与MRP之间无相关性。但同时表达时 ,多数发展成为难治性白血病。结论　儿童急性白血病的发生、发展、转归与WT1、MRP过度表达有关。二者均可作为临床指导治疗和判断预后的指标。     

WT1基因在急性髓细胞性白血病微小残留病检测中的应用

目的探讨联合检测肾母细胞瘤基因1(WT1)和融合基因在急性髓细胞性白血病(AML)微小残留病(MRD)中的应用价值。方法收集2018年1月至2020年12月于该院确诊并完成RUNX1-RUNX1T1、早幼粒细胞白血病蛋白(PML)/视黄酸受体(RAR)α和WT1基因检测的伴重现性遗传学异常的AML患者。检测RUNX1-RUNX1T1、PML/RARα和WT1转录本水平,将RUNX1-RUNX1T1、PML/RARα融合基因转录本水平为0%看作融合基因阴性组,将RUNX1-RUNX1T1或PML/RARα融合基因转录本水平均不为0%看作融合基因阳性组。结果融合基因阴性组患者WT1转录本水平为0.096%(0.007%,1.990%),融合基因阳性组患者WT1转录本水平为1.420%(0.100%,60.340%),融合基因阴性组患者WT1转录本水平低于融合基因阳性组患者(P<0.05)。融合基因高表达组(融合基因转录本水平≥1%)患者融合基因转录本水平高于WT1转录本水平(P<0.05),融合基因低表达组(融合基因转录本水平<1%)患者WT1转录本水平高于融合基因转录本水平(P<0.05)。结论可联合检测融合基因和WT1,以此提高MRD的检测灵敏度。     

Genetic polymorphism of cytochrome P450 1A1 (Cyp1A1) and glutathione transferases (M1, T1 and P1) among Africans.

The co-ordinate expression and regulation of the drug metabolising enzymes, cytochrome P4501A1 (CYPlAl) and glutathione transferases (GSTM1, GSTT1 and GSTP1), and their metabolic balance in the cells of target organs may determine whether exposure to carcinogens results in cancer. Besides showing variability in activity due to induction and inhibition, these enzymes also exhibit genetic polymorphism that alter enzyme levels and activity. We determined frequencies of common allelic variants of CYP1A1 and glutathione (M1, T1 and P1) among Tanzanians, South African Venda and Zimbabweans using PCR/restriction fragment length polymorphism techniques. The CYP1A1 Val462 mutant variant was found at a frequency of 1.3% among 114 subjects. The GSTM1*0 genotype was found at a frequency of 29% and 33% among Tanzanian psychiatric patients and healthy volunteers, respectively. Similarly, the GSTT1*0 polymorphism was present with a frequency of 25% in both the psychiatric patients and healthy controls. The frequency of GSTP1 Val105 variant was 16%, 12% and 21% among Tanzanians, South African Venda and Zimbabweans, respectively. We conclude here that CYP1A1 Val462 polymorphism is very rare among Africans. This is the first report of the GSTP1 Val105 variant frequency in African populations. We show here that there are no differences in frequencies of the variant alleles for CYP1A1, GSTM1, GSTT1 and GSTP1 in the three African populations.     

子宫内膜癌中CYP1A1，CYP1B1及COMT mRNA的表达及临床意义

[目的]探讨细胞色素P450酶（CYP）1A1, 1B1和儿茶酚-O-甲基转移酶（COMT） mRNA的表达与子宫内膜癌发生发展的关系.[方法]采用RT-PCR方法检测48例子宫内膜癌患者和53例健康对照个体的外周血淋巴细胞中CYP1A1,CYP1B1和COMT mRNA的表达.[结果]子宫内膜癌患者外周血淋巴细胞中CYP1A1,CYP1B1 mRNA的表达明显增高,经Logistic回归分析后, CYP1B1 mRNA的高表达能增加子宫内膜癌的发病风险,进一步分析显示Ⅱ期子宫内膜癌CYP1B1 mRNA的表达高于Ⅰ期患者,差异具有显著性意义.[结论]外周血淋巴细胞中CYP1B1 mRNA的高表达与子宫内膜癌的发生发展密切相关.     

Vectorial transport of enalapril by Oatp1a1/Mrp2 and OATP1B1 and OATP1B3/MRP2 in rat and human livers

Although Oatp1a1 (rat organic anion-transporting polypeptide 1a1) was the transporter found responsible for the hepatocellular entry of enalapril (EN) into the rat liver, the canalicular transporter involved for excretion of EN and the metabolite, enalaprilat (ENA), was unknown. The Eisai hyperbilirubinemic rat (EHBR) that lacks Mrp2 (multidrug resistance-associated protein 2) was used to appraise the role of Mrp2 in the excretion of [3H]EN and its metabolite [3H]ENA in single-pass rat liver preparations. Although the total and metabolic clearances and hepatic extraction ratios at steady-state were virtually unaltered for EN in EHBR compared with published values of Sprague-Dawley rats, the biliary clearances of EN and ENA were significantly reduced almost to zero (P<0.05). Involvement of human OATP1B1, OATP1B3, and MRP2 in EN transport was further assessed in single- or double-transfected mammalian cells. Human embryonic kidney 293 cells that expressed OATP1B1 or OATP1B3 showed that OATP1B3 transport of EN (20-500 microM) was of low affinity, whereas transport of EN by OATP1B1 was associated with the Km of 262+/-35 microM, a value similar to that for Oatp1a1 (214 microM). The transcellular transport of EN via human OATP1B1 and MRP2, investigated with the double-transfected Madin-Darby canine kidney (MDCK) II cells in the Transwell system, showed that the sinusoidal to canalicular flux of EN in the OATP1B1/MRP2/MDCK cells was significantly higher (P<0.05) than that of mock/MDCK and OATP1B1/MDCK cells. EN was transported by Oatp1a1 and Mrp2 in rats and OATP1B1/OATP1B3 and MRP2 in humans.     

Baicalein inhibits DMBA-DNA adduct formation by modulating CYP1A1 and CYP1B1 activities.

Flavonoids are phenolic compounds isolated from plants, and several of them like genistein and quercetin, have been documented to be effective in preventing cancer. Baicalein, a flavonoid extracted from the root of Scutellaria species, is widely used as a health supplement and herbal medicine in Asian countries. In this study, the chemopreventive effect of baicalein on 7,12-dimethylbenz[a]anthracene (DMBA)-induced DNA damage was evaluated in an established cell culture model. In a preliminary screening, baicalein was identified to be a strong inhibitor to EROD activities induced by DMBA in MCF-7 cells. Subsequent enzyme kinetic analysis revealed that baicalein was a competitive inhibitor to EROD, and CYP1A1 and CYP1B1 gene expressions were also determined. Baicalein could reduce the CYP1A1/1B1 mRNA expressions induced by DMBA, and the mRNA abundance of CYP1A1 appeared to be more responsive than that of CYP1B1. A XRE-luciferase gene reporter assay indicated that AhR transactivation was suppressed. Since CYP1A1/1B1 were responsible for the biotransformation of polycyclic aromatic hydrocarbons, baicalein also demonstrated its ability to reduce DMBA-DNA adduct formation in MCF-7 cells. This study suggested that the natural occurring baicalein could be an agent preventing carcinogen-DNA adduct formation.     

Urinary transforming growth factor-beta1 and alpha1-microglobulin in children and adolescents with type 1 diabetes

OBJECTIVE: Transforming growth factor (TGF)-beta1 is an important mediator in the pathogenesis of diabetic nephropathy. Urinary TGF-beta1 reflects TGF-beta1 production in the kidney, and alpha1-microglobulin tubular dysfunction. These 2 markers were studied in the early phases of type 1 diabetes. RESEARCH DESIGN AND METHODS: There were 113 type 1 diabetic children and adolescents (mean +/- SD: age 14.1 +/- 2.9 years, and diabetes duration 7.4 +/- 2.9 years, HbA1c 9.3 +/- 1.5%) and 39 healthy subjects (age 13.8 +/- 2.8 years) who participated in the study. Of the diabetic patients, 105 were normoalbuminuric (2-3 consecutive overnight urinary albumin excretion rates [AERs] <20 microg/min) and 8 had microalbuminuria (at least 2 AERs 20-200 microg/min). Overnight urinary TGF-beta1 and alpha1-microglobulin levels were measured and the results expressed as the ratio to urinary creatinine concentration. RESULTS: Data are medians (range). Diabetic patients had higher urinary TGF-beta1 levels than those of control subjects: 0.9 ng/mg (0.05-122.3) vs. 0.3 ng/mg (0.05-2.2) creatinine, respectively (P = 0.003). Urinary TGF-beta1 levels correlated with urinary glucose (r = 0.2, P = 0.03) and alpha1-microglobulin (r = 0.2, P = 0.02) levels, but not with HbA1c, AER, age, or duration of diabetes. In 43 patients with urinary TGF-beta1 above the control levels, urinary TGF-beta1 levels correlated with urinary glucose (r = 0.6, P < 0.001) and alpha1-microglobulin (r = 0.6, P < 0.001) levels. Diabetic patients had higher urinary alpha1-microglobulin levels than those of control subjects: 4.8 microg/mg (0.6-48.8) vs. 2.7 microg/mg (0.8-11.6) creatinine, respectively (P < 0.001). Alpha1-microglobulin levels correlated with AER (r = 0.2, P = 0.02), HbA1c (r = 0.3, P = 0.001), urinary glucose (r = 0.5, P < 0.001), and urinary TGF-beta1 levels. CONCLUSIONS: An early rise in urinary TGF-beta1 levels was observed in young type 1 diabetic patients. Urinary TGF-beta1 is associated with 2 interrelated tubular markers, alpha1-microglobulin and urinary glucose.     

周期蛋白D_1,B_1在子宫颈癌中的表达及其与临床病理间的关系

目的探讨CyclinD1,CyclinB1在子宫颈癌中的表达及其与临床病理间的关系。方法收集哈尔滨医科大学附属四院收治的38例子宫颈癌组织。采用S-P免疫组织化学法,选用CyclinD1,CyclinB1多克隆抗体,分别检测其在38例子宫颈癌中的表达率。结果 CyclinD1在38例子宫颈癌中的阳性表达率为76.32%,CyclinB1的阳性表达率为52.63%,CyclinD1的表达率高于CyclinB1,并且其表达差异有统计学意义(经Fisher确切概率法,P<0.05)。CyclinD1,CyclinB1两者的阳性表达率与淋巴结有无转移之间的关系:Cy-clinD1高于CyclinB1,但差异无统计学意义(经Fisher确切概率法,P>0.05)。结论 CyclinD1驱动细胞由G1期进入S期,促进细胞增殖,决定细胞周期进程的速度和方向,在子宫颈癌的发展过程中起着主要的作用。CyclinB1是在有丝分裂过程中起调控作用的周期蛋白,受着cyclinD1作用的影响。