应用实时定量PCR技术检测慢性粒细胞白血病患者p210~(bcr/abl)转录本的临床意义

目的 :研究 p2 1 0 bcr/abl融合基因转录本在慢性粒细胞白血病 ( CML)不同病程阶段的表达水平及其在病情进展中的意义。方法 :应用实时定量PCR( RQ- PCR)技术检测 68例CML患者骨髓单个核细胞中的 p2 1 0 bcr/abl转录本 ,并将其含量以看家基因 ABL表达量进行标化。结果 :本组中 p2 1 0 bcr/abl转录本绝对含量为 4.0 1× 1 0 2～ 8.90× 1 0 5拷贝 /5 0 ng(中位 1 .0 7× 1 0 5拷贝 /5 0 ng) ,ABL纠正后的相对含量为 0 .0 1～ 48.67(中位 5 .0 3)。加速期和急变期患者的p2 1 0 bcr/abl转录本水平较慢性期患者明显增高 (中位含量分别为 9.0 4、7.36和 3.5 3) ;干扰素治疗无效者较慢性期患者的 p2 1 0 bcr/abl水平则明显增高。结论 :p2 1 0 bcr/abl转录本过度表达可能与 CML病情进展相关 ,RQ- PCR可用于对 CML的病情监测。     

干扰素α-2b治疗慢性粒细胞性白血病的前瞻性随机对照研究

目的比较高、低剂量干扰素(IFN)α2b治疗慢性粒细胞性白血病(CML)的效果。方法建立检测融合基因bcr abl的荧光实时定量逆转录聚合酶链反应(RQ PCR)方法,观察治疗后融合基因表达水平的变化。选择30例临床初诊的CML患者随机分为两组,先服用羟基脲控制外周血白细胞达20×109/L以下,然后分别给予IFNα2b300万IU隔日皮下注射(3MIU组)和500万IU每周6次(5MIU组)皮下注射,治疗3～6个月,每月抽取骨髓标本,检测融合基因bcr abl的表达情况。结果RQ PCR的灵敏度达50拷贝bcr abl;分别取1×103拷贝/μl和1×107拷贝/μlbcr abl质粒,以及1例CML患者的cDNA同时作8管平行扩增bcr abl融合基因,批内变异系数分别为2.35%、1.48%和1.17%,不同批次之间以K562细胞cDNA作重复性分析,批间变异系数为5.13%;CML初诊患者bcr abl/GAPDH水平介于0.010～5.799,中位值为0.098;治疗3个月后3MIU组和5MIU组患者骨髓bcr abl水平平均下降19%和24%,两组比较差异无统计学意义(P=0.398),但是3MIU组副作用相对较小。结论RQ PCR监测融合基因bcr abl表达可以有效观察CML患者使用IFN的治疗效果;不同CML患者白血病细胞的bcr abl表达水平有较大差异;隔日3MIUIFN皮下注射治疗即可有效抑制CML白血病细胞的增殖,且副作用较小。     

格列卫对慢性粒细胞白血病病人Ph染色体和bcr／abl融合基因表达影响

目的 探讨格列卫治疗Ph染色体阳性慢性粒细胞白血病病人后骨髓Ph染色体及bcr/abl融合基因表达的变化.方法 采用R显带和半定量RT-PCR分别检测11例慢性粒细胞白血病病人服用格列卫前后骨髓标本中Ph 染色体和bcr/abl基因的表达.结果 用药后9例慢性期慢性粒细胞白血病病人,7例分别在用药3～24个月后染色体检查为Ph染色体阴性,1例用药1个月后其Ph染色体仍为阳性,后失访;1例用药2个月后 Ph染色体阳性率为30%;1例急变期慢性粒细胞白血病病人用药6个月后其Ph染色体阳性率为70%;1例加速期慢性粒细胞白血病病人用药12个月后Ph染色体阳性率为60%.服用格列卫的CML病人Ph染色体转阴后仍能检测到bcr/abl基因的表达,但其表达水平较用药前明显降低,差异有显著性(t=3.77、5.13,P<0.01),随着用药时间的延长bcr/abl基因表达逐渐降低.结论 格列卫能有效治疗Ph染色体阳性的慢性粒细胞白血病病人,并且对其预后具有重要的意义.     

bcr/abl融合基因在慢性粒细胞白血病中的表达及临床意义

目的 探讨bcr/abl融合基因的表达在慢性粒细胞白血病(CML)中的诊断、疗效及预后观察中的意义.方法 采用荧光定量逆转录多聚酶链反应(FQ -PCR)技术,对8例复治CML患者和作为对照组的2例急性非淋巴细胞性白血病患者的骨髓或外周血进行bcr/abl融合基因检测;同时比较患者的血象、骨髓象等相关实验室指标.结果 CML中7例慢性期(CP)和1例急变期(BC)患者,均不同程度地表达了bcr/abl融合基因,而对照组该融合基因的表达为阴性.结论 bcr/abl基因是CML发病的分子基础,定量检测bcr/abl融合基因对于CML的诊断、临床分型、疗效观察及预后判断有重要意义.     

In Situ-RT PCR定量检测16例慢性粒细胞白血病bcr/abl融合基因

目的：研究原位逆转录多聚酶链反应技术（In Situ-RT PCR)在检测慢性粒细胞白血病（CML）bcr/abl融合基因中的应用价值及其意义。方法：用In Situ-RT PCR方法测定CML病人bcr/abl融合基因表达量,结果：在不破坏完整性条件下细胞系K552和CML病人检测到bcr/abl mRNA的扩增产物,表现为细胞浆内出现蓝紫色颗粒,计算这种阳 细胞的百分率来相对定量地,CML病人bcl/abl mRNA的表达水平,这种检测技术的敏感性达10^-4水平以上,结论：In Situ-RT PCR可以相对定量地检测CML病人bcl/abl mRNA的表达,直观地评价α－干扰素（α－IFN）,骨髓移植（BMT）等治疗CML的疗效,本检测方法敏感性高,又可用做微小残留病（MRD）的检测。αα     

酪氨酸蛋白磷酸酶2高表达对p210bcr/abl诱导的慢性粒细胞白血病细胞增殖与凋亡抵抗的影响

目的研究Src癌基因同源的酪氨酸蛋白磷酸酶2(Shp2)在慢性粒细胞白血病(CML)细胞中的表达,及其在p210bcr/abl诱导的白血病细胞恶性增殖和凋亡抵抗中的作用。方法收集25例p210bcr/abl阳性CML患者白血病细胞样本,8例非肿瘤病人骨髓和10例正常人外周血细胞样本作阴性对照,K562和KU812白血病细胞系作为p210bcr/abl阳性对照,KG1白血病细胞作为Shp2阳性对照。用Western印迹技术定量分析与比较Shp2在白血病细胞和正常骨髓造血细胞中的表达情况,通过特异性抑制剂分别下调Shp2和白血病融合基因p210bcr/abl后,观察Shp2表达对p210bcr/abl阳性白血病细胞增殖与凋亡的作用。细胞增殖与凋亡检测应用流式细胞仪。结果(1)磷酸化Shp2蛋白在92%患者CML白血病细胞样本中呈高表达状态,而在8例非肿瘤患者骨髓和10例正常人外周血细胞中低表达或不表达。磷酸化Shp2/β肌动蛋白比值分别为0.91±0.62、0.16±0.09和0.03±0.05(P均<0.01)。(2)下调Shp2蛋白表达后,白血病细胞凋亡率从4.89%上升到38.69%(P<0.01),而S期细胞从33.6%下降到10.8%(P<0.01)。(3)特异性抑制p210bcr/abl融合基因表达蛋白后,白血病细胞中磷酸化Shp2蛋白明显下降,同时出现明显细胞凋亡与生长抑制现象。结论(1)磷酸化Shp2蛋白在慢性粒细胞白血病细胞患者中呈过度表达状态,并且与白血病细胞恶性增殖与凋亡抵抗密切相关。(2)bcr/abl融合基因阳性患者的白血病细胞中Shp2的过度表达可能由p210bcr/abl蛋白激活引起。     

血小板显著增多为首发表现的慢性髓细胞白血病2例并文献复习

目的探讨以血小板显著增多为首发表现的慢性粒细胞白血病（CML）的临床及分子生物学特点。方法应用细胞涂片观察形态学;RT-PCR检测bcr/abl融合基因;常规染色体检测细胞遗传学变化。结果血小板显著增多为首发表现的CML是一组具有独特临床和生物学特点的疾病,表达bcr/abl融合基因,可进一步向CML加速期、急变期演变,应早期进行积极干预。结论对血小板显著增多的患者应及时进行Ph染色体及bcr/abl融合基因表达水平的检测,及早鉴别原发性血小板增多症（ET）及慢性粒细胞性白血病（CML）,避免误诊、误治。     

初发慢性髓系白血病bcr/abl融合基因转录子与临床关系的观察

目的　研究初发慢性髓系白血病bcr/abl融合基因转录子类型与临床关系。方法　使用RT PCR对 5 1例初发CML患者进行bcr/abl融合基因转录子检测。结果　b3a2型患者血小板数高于b2a2型患者 ,但b3a2型患者WBC数低于b2a2型患者 (P <0 .0 5 ) ;血红蛋白浓度与b2a2型患者比较无差异 (P >0 .0 5 ) ;b3a2型和b2a2型患者比较AKP和Ph1染色体比例无差异 (P >0 .0 5 ) ;M bcr/abl阳性患者伴m bcr/abl阳性率为 47 0 % ,其中b3a2型患者有m bcr/abl融合基因转录子 14例 ,b2a2型患者有 10例。结论　b3a2型CML患者初发时更易有血小板升高表现 ,但b2a2型患者更易有WBC数升高的表现 ,较多的初发CML患者同时伴有m bcr/abl阳性。     

ES探针荧光原位杂交法检测bcr/abl融合基因

目的探讨应用ES型bcr/abl探针(双色单融合)荧光原位杂交技术(FISH)在检测bcr/abl融合基因中的价值。方法慢性髓细胞性白血病(CML)初诊或复诊患者30例(骨髓标本43份),行普通染色体核型分析和ES型bcr/abl探针荧光原位杂交分析。结果 43份骨髓标本,6份未做骨髓培养,6份培养失败,在其余31份培养成功的标本中,找到Ph1染色体9份,另有1份标本多次检查都发现存在21p+染色体异常;43份骨髓都经FISH检测,13份标本发现bcr/abl融合基因。结论 ES型bcr/abl探针对CML初诊病例的bcr/abl融合基因有较高的检出率;对复发病例,不仅能辅助诊断CML,而且在监测白血病微小残留病灶、判断疾病的进展上均有较高的灵敏度。     

慢性粒细胞性白血病bcr/abl融合基因的表达

目的：研究慢性粒细胞性白血病患者bcr／abl融合基因阳性率及其临床意义。方法：取慢性粒细胞性白血病(CML)患者骨髓或外周血，提取单核细胞的总RNA，用逆转录多聚酶链反应(RT-PCR)，观察其ber／abl基因的表达。结果：9例CML患者中，8例ber／abl基因阳性，占88.9％。结论：bcr／abl基因检测可以用来辅助CML的临床诊断。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.