降落式PCR和SSCP检测淋巴细胞白血病单克隆T细胞受体γ基因重排

目的 以降落式PCR和单链构型多态性(SSCP)方法检测淋巴细胞白血病单克隆T细胞受体(TCR)γ基因重排，探讨其在淋巴细胞白血病诊断中的价值。方法 盐析法提取淋巴细胞白血病患者外周血白细胞DNA，TCR-γ基因重排通用引物和降落式PCR扩增基因重排。分别采用琼脂糖凝胶电泳、SSCP和DNA直接测序方法检测PCR扩增产物。阳性细胞系DNA模板与反应增生性淋巴组织DNA按不同比例混合后扩增，检测降落式PCR的灵敏度。结果 琼脂糖电泳中，18例T淋巴细胞白血病中有15例、4例B淋巴细胞白血病中有2例为阳性。阳性标本经SSCP进一步分析，呈不连续条带。DNA直接测序证实扩增产物为TCR-γ基因重排。阳性对照模板占1％以上时可得到阳性扩增结果。结论 TCR-γ基因重排通用引物结合降落式PCR可有效扩增T淋巴细胞白血病基因重排，可用于T淋巴细胞白血病的辅助诊断。     

应用特异性基因标志早期诊断中枢神经系统白血病的研究

目的探讨特异性基因标志对急性淋巴细胞白血病(ALL)患中枢神经系统白血病(CNSL)早期诊断的价值。方法 用聚合酶链反应(PCR)技术扩增ALL患不同期脑脊液免疫球蛋白重链(IgH)基因重排和T细胞受体T(TCR7)基因重排。结果①12例ALL脑脊液标本两种PCR基因扩增6例(9份)获阳性结果,与临床结果相符。②有2例患临床不符CNSL,但PCR阳性,经观察治疗后得到确诊。结论本法特异性强．且有较高的灵敏度,可为ALL患CNSL早期诊断的有效方法之一。     

基因重排检测技术的建立及其在淋巴瘤诊断中的应用

目的:建立基于克隆性分析技术的B和T淋巴细胞产生的抗原受体分子基因重排检测技术,探讨B淋巴细胞免疫球蛋白及T淋巴细胞受体基因重排在恶性淋巴瘤临床诊断中的应用. 方法: 从组织蜡块中切取组织片5～10张,提取基因组DNA,PCR扩增,聚丙烯酰胺凝胶电泳后银染显示基因重排结果. 结果:20例良性淋巴组织反应性增生病例中, 淋巴细胞IgH及TCR基因重排均为阴性;18例T细胞非何杰金氏淋巴瘤中TCRγ基因重排12例, TCRβ基因重排3例,未见有IgH基因重排;32例B细胞性非何杰金氏淋巴瘤标本中有28例IgH基因重排阳性,有2例TCRγ和IgH基因重排均阳性;3例未确诊的疑难病例均为IgH基因重排;IgH及TCR基因重排检出率在淋巴瘤组与良性组之间,差异有显著意义(P<0.05). 结论:应用PCR法检测B淋巴细胞IgH和T淋巴细胞TCR基因重排可作为淋巴细胞来源的良、恶性病变的一种辅助诊断手段,且适用于石蜡标本以进行回顾性研究.     

降落式PCR和SSCP检测淋巴细胞白血病单克隆T细胞受体γ基因重排

目的 以降落式PCR和单链构型多态性(SSCP)方法检测淋巴细胞白血病单克隆T细胞受体(TCR)γ基因重排,探讨其在淋巴细胞白血病诊断中的价值。方法 盐析法提取淋巴细胞白血病患者外周血白细胞DNA,TCR-γ基因重排通用引物和降落式PCR扩增基因重排。分别采用琼脂糖凝胶电泳、SSCP和DNA直接测序方法检测PCR扩增产物。阳性细胞系DNA模板与反应增生性淋巴组织DNA按不同比例混合后扩增,检测降落式PCR的灵敏度。结果 琼脂糖电泳中,18例T淋巴细胞白血病中有15例、4例B淋巴细胞白血病中有2例为阳性,阳性标本经SSCP进一步分析,呈不连续条带。DNA直接测序证实扩增产物为TCR-γ基因重排。阳性对照模板占1%以上时可得到阳性扩增结果。结论 TCR-γ基因重排通用引物结合降落式PCR可有效扩增T淋巴细胞白血病基因重排,可用于T淋巴细胞白血病的辅助诊断。     

克隆演化与中枢神经系统白血病复发的关系

应用聚合酶链反应（ＰＣＲ）技术检测４５份急性淋巴细胞白血病（ＡＬＬ）脑脊液标本ＩｇＨ、ＴＣＲＶγＩ－Ｊγ及ＴＣＲＶδ２－Ｄδ３基因重排，从２２份标本（４８．９％）发现基因重排阳性，而用形态学方法检查，阳性率为２４．４％（１１／４５）；２２例阳性病人中的２例（９．１％）脑脊液白血病细胞基因重排类型同骨髓标本不一致，此两例均为中枢神经系统白血病（ＣＮＳＬ）复发。上述结果表明，应用ＰＣＲ技术检测ＡＬＬ脑脊液标本重排基因有助于隐匿ＣＮＳＬ诊断；克隆演化现象可能是某些单独ＣＮＳＬ复发的主要原因     

多聚酶链反应检测淋巴细胞白血病免疫球蛋白重链基因重排及其临床意义

用多聚酶链反应技术扩增42例急、慢性白血病患骨髓白血病球蛋白重链（IgH)基因重排产生的第三互补决定区（CDR-Ⅲ）DNA序列。结果显示：35例淋巴细胞白血病中25例出现IgH基因单克隆性重排（71.4%)，其中急、慢性B淋巴细胞白血病各为80%（16/20）和87.5%(7/8)及2例T、B淋巴细胞双表型白血病，另外有1例急性未分化白敌国病也出现这种重排，而5例，急性T淋巴细胞白血病和6例急性髓细胞白血病均无重排，对3例CDR-Ⅲ阳性克隆的B-ALL病例在完全缓解期进行微小残留病变检测，其中2例PCR阳性病人先后复发，1例PCR阴性病人缓解持续时间明显长于阳性病人。     

老年人急性白血病T细胞受体VγI-Jγ基因重排分析

目的为研究老年人急性白血病分子生物学特点及临床意义。方法提取34例老年人急性非淋巴细胞白血病(ANLL)、9例老年人急性淋巴细胞白血病(ALL)及10名正常献血员DNA标本,应用PCR技术扩增TCRVγI-Jγ基因重排,对阳性的PCR产物进行SSCP分析。结果34例老年人ANLL中有16例(47.1%)发现TCRVγI-Jγ基因重排,老年人ANLL TCRVγI-Jγ基因重排检测阳性率显著高于非老年人ANLL组(P<0.025)。9例老年人ALL中有7例(77.8%)发现TCRVγI-Jγ基因重排,其中3例(33.3%)存在寡/亚克隆重排,老年人ALL寡/亚克隆重排阳性率显著高于非老年人ALL组(P<0.05)。10例正常献血员PCR技术扩增TCRVγI-Jγ基因重排均为阴性。TCRVγI-Jγ基因重排阳性老年人ANLL完全缓解率为25.0%(4/16),显著低于重排阴性老年人ANLL组(61.1%)(P<0.05)。结论应用PCR/SSCP方法分析老年人急性白血病TCRγ重排可作为疗效和预后判断的新指标。老年人急性白血病寡/亚克隆重排检测阳性率显著高于非老年人组可能是导致其疗效差的一个重要因素。     

儿童急性淋巴细胞性白血病IgH基因和TCR基因重排的研究

采用PCR或巢式PCR方法对71例未治疗的急性淋巴细胞性白血病（ALL）患儿骨髓样本的免疫球蛋白重链基因（IgH）和T细胞受体基因（TCR）进行了克隆性重排检测，结果显示在52例B系ALL及19例T系ALL中分别存在82．7％（43例）和15．8％（3例）的IgHCDRⅢ区域重排，而TCRVδ2Dδ3基因重排见于32．7％的B系ALL（17例）和63．2％的T系ALL（12例），二种基因重排在儿童ALL的覆盖面达到88．7％（63例），此结果提示IgHCDRⅢ和TCRVδ2Dδ3基因可作为监测儿童ALL的标志基因；采有相同方法对完全缓解期患儿的67份脑脊液标本进行了检测，结果显示9例存在IgHCDRⅢ基因重排，4例存在TCRVδ2Dδ3基因重排（与骨髓标本重排类型一致），提示采用PCR技术对ALL患儿CSF标本进行IgH和Vδ2Dδ3基因重排的检测对早期诊断中枢神经系统白血病（CNSL）具有较大价值；采用有限稀释法对完全缓解期的儿童ALL病例骨髓样本进行上述二种基因重排的定量分析，结果提示对儿童ALL进行微小残留病（MRD）的持续追踪监测可指导化疗方法的选择，观察治疗效果并预测预后。     

多聚酶链反应检测急淋白血病微量残留病灶及其临床应用

应用多聚酶链反应 (PCR)扩增IgH重排产生的CDR -Ⅲ序列和TcRr基因的重排序列 ,检测 2 2例急淋白血病 2 3份骨髓标本 ,其中 1例初治急淋白血病标本检出IgH重排和TcRr重排 ,2 1例获缓解的患儿骨髓标本有 4例检出残留白血病细胞。阳性病例在 1周至半年不等出现白血病复发或死亡。本方法MRLC检测敏感性为 10 -4 。     

多重聚合酶链反应技术与流式细胞术在B细胞非霍奇金淋巴瘤骨髓侵犯的鉴别诊断比较分析

目的探讨多重聚合酶链反应(PCR)技术检测基因重排及流式细胞术检测淋巴细胞表面标志物在非霍奇金淋巴瘤诊断中的应用。方法采用37例B细胞非霍奇金淋巴瘤(B-NHL)骨髓侵犯患者骨髓标本作为分析对象,20例反应性淋巴组织增生患者骨髓标本为对照,同时采用流式细胞术和多重PCR技术检测,对结果进行分析,并与病理诊断及免疫组织化学结果进行比较,探讨两者的应用价值。结果在37例病理确诊为B-NHL患者骨髓的骨髓标本,PCR检测结果 36例阳性,1例阴性;流式细胞术检测结果 34例阳性,3例阴性。结论多重PCR技术与流式细胞术检测骨髓标本在B-NHL鉴别诊断中具有一定的应用价值,多重PCR技术应用价值高于流式细胞术。     

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.