多药耐药基因TopoⅡ仪mRNA在胃癌及其相邻正常组织中表达

目的 :研究拓朴异构酶TopoⅡα在胃癌内在耐药性中的作用。 方法 :采用RT PCR方法 ,检测 36例胃癌及其相邻正常组织中TopoⅡαmRNA的表达状态。结果 :TopoⅡα在胃癌中表达 (47.2 % ) ,显著高于其相邻正常组织 (13.9% ,P <0 .0 5 )。结论 :胃癌存在内在性耐药 ,TopoⅡα表达是胃癌内在性耐药的影响因素之一     

HL60/VCR白血病细胞多药耐药机制的初步探讨

目的探讨白血病细胞产生耐药的机制,以及VEGF在白血病细胞产生耐药过程中的作用。方法采用反复暴露并逐渐提高诱导药物浓度的方法建立HL60/VCR多药耐药细胞系,MTT法检测细胞的耐药性及交叉耐药性;RT-PCR检测诱导前后mdr1、MRP、TopoⅡα、GSTπ和VEGF mRNA的表达情况。结果诱导后的HL60/VCR耐药细胞不仅对VCR高度耐药,对其他类化疗药物亦具有交叉耐药性;与HL60相比,HL60/VCR的mdr1、TopoⅡα和VEGF mRNA的表达明显增强,GSTπmRNA的表达无明显变化;HL60和HL60/VCR均不表达MRP mRNA。结论耐药基因mdr1和TopoⅡα以及VEGF共同参与HL60/VCR多药耐药的形成。     

多耐药基因在乳腺癌组织中表达及临床意义

目的全面了解乳腺癌内源性耐药情况,实现肿瘤病人个体化化疗。方法应用荧光定量PCR方法检测51例术前未经化疗的乳腺癌和癌旁组织中mdr1、MRP、LRP、GST-π、TOPOⅡα、BCRP基因表达。结果这6个多药耐药基因在乳腺癌组织和癌旁组织中的阳性表达率均有显著统计学意义;mdr1、LRP、GST-π、TOPOⅡα在乳腺癌组织和癌旁组织之间的表达水平无显著统计学意义（P〉0.05）;MRP和BCRP在乳腺癌组织中的表达水平显著高于其在癌旁组织中的表达水平,差异有统计学意义（P〈0.05）。结论 MRP和BCRP可能与乳腺癌的内源性耐药有关,mdr1、LRP、GST-π、TOPOⅡα的表达可能与乳腺癌的内源性耐药无关,乳腺癌患者进行术前的新辅助化疗是可行的。     

32例肝癌组织中多药耐药基因及其产物P-糖蛋白的实验研究

探讨肝癌组织中mdr－1（多药耐药基因）mRNA及P－170（P－糖蛋白）的表达水平，采用RT－PCR及免疫组织化学SP法检测32例肝癌组织中mdr－1mRNA和P－糖蛋白的表达．发现不仅肝癌组织中mdr－1和P－糖蛋白的表达显著高于对照组，而且转移组中P－糖蛋白的表达高于非转移组．提示检测mdr－1／P－gp对肝癌制订治疗方案，判断疗效及MDR逆转剂的应用具有重要的临床意义；mdr－1mRNA／P－gp的过表达可能与肝肿瘤的转移预后有关．     

脊索瘤细胞系CM-319中放化疗抵抗相关基因的表达及临床意义

[目的]探讨多药耐药相关基因如缺氧诱导因子1d（HIF-1α）、多药耐药基因（MDR1）及多药耐药相关蛋白（MRP1）在脊索瘤细胞系CM－319中的表达及临床意义。[方法]用RT—PCR检测HIF－1α、MDR1、MRP1基因表达,间接免疫荧光及免疫细胞化学Envision法检测HIF－1α、MRPl蛋白在CM－319中的表达情况,并分析其与放化疗抵抗的关系。[结果]常氧条件下,在脊索瘤细胞系中能检测到HIF～1α及MRP1 mRNA的表达,但未检测到MDR1 mRNA的表达（P〈0．01）,间接免疫荧光及免疫细胞化学可以检测到HIF-1α表达在细胞浆及核中,而MRP1主要表达在细胞浆中。[结论]HIF-1α与MRP1的过度表达在脊索瘤的放化疗抵抗中可能具有重要作用,而MDR1表达未被检测到,提示脊索瘤的放化疗抵抗主要与HIF－1α和MRP1相关。     

三种肿瘤耐药基因在非小细胞肺癌中的表达及临床意义

目的探讨肺耐药蛋白（Lung resistance protein，LRP），多药耐药蛋白（Multldrug resistance—associated protein，MRP）和多药耐药基因（Multidrug resistance，MDR1）的mRNA在非小细胞肺癌（Non-small cell cancer，NSCLC）中共表达及临床意义。方法RT—PCR检测NSCLc冰冻组织中上述耐药基因的mRNA水平。结果LRP、MRP、MDR1mRNA阳性率分别为71．2％、81．8％和37．9％；MRP与、MDR1、MRP与LRP、LRPYUMDR1及MRP、LRPYUMDR1共表达者分别为16．7％（11／66）、53．0％（35／66）、19．7％（13／66）及12．1％（8／66）。LRP与MRP阳性相关（t=0．47，P=0．001），LRP、MRP与MDR1无相关性（r=0．98，P=0．0r77；r=0．25，P=0．053）．3种耐药基因表达在NSCLC各期、各分化组中差异无显著性（P≥0．05）。肺腺癌换了有效组LRP、MBPmRNA（0．53±0．08；0．41±0．01）及鳞癌化疗有效组LRPmRNA表达（0．42±0．03）明显低于换了无效组（1．14±0．3，0．82±0．04，0．97±0．01，P〈0．05）；LRP与MRP、LRP、MRP与MDR1共表达者，中位生存期明显缩短（P〈0．05）。多变量Cox回归分析，仅LRP＋MRP、LRP＋MRP＋MDR1共表达有预后意义。结论非小细胞肺癌原发耐药与LRP、MRP、MDR1共同作用有关。     

原发性肾癌组织中MDR1和MRP基因的表达

①目的　研究肾癌组织中多药耐药 (MDR1)基因和多药耐药相关蛋白 (MRP)基因的mRNA表达及其临床意义。②方法　应用定量逆转录聚合酶链反应 (RT PCR) ,检测 2 0例原发性肾癌组织、10例正常人肾脏组织新鲜标本MDR1和MRP基因的mRNA表达水平。③结果　正常肾脏组织和肾癌组织均有MDR1和MRP基因表达 ,肾癌组织MDR1基因表达水平低于正常肾脏组织 ,MRP基因表达水平高于正常肾脏组织 ,差异有显著性 (t=2 .2 93,2 .96 8,P <0 .0 5 ,0 .0 1)。MDR1和MRP基因的表达与肾癌的病理类型、病理分期和组织学分级无关 ,与肾癌病人的无进展生存期无关 ,正常肾脏组织的MDR1基因和MRP基因的表达呈高度正相关关系 (r =0 .932 ,P<0 .0 1) ;肾癌组织MDR1基因和MRP基因的表达无明显相关关系。④结论　MDR1基因和MRP基因复合表达与肾癌固有的多药耐药现象有关     

MDR_1mRNA检测在胃癌化疗方案选择中的意义(附20例分析)

本文对20例胃癌细胞原发性MDR1mRNA水平进行检测，并且对其影响肿瘤细胞mdr－1基因表达的因素采用多因素电子计算机Logistic回归模型进行分析。结果显示：胃癌细胞MDR1mRNA表达与年龄、性别、肿瘤的大小、分期、分化程度等因素之间无统计学关系（P＞0．1）；与肿瘤的淋巴结转移（P＝0．0107）、局部组织浸润（P＝0．0009）关系密切。提示：胃癌病人术后化疗方案的设计应根据肿瘤细胞mdr－1基因检测结果而拟定。对术中肿瘤已有淋巴结转移和（或）已有局部组织浸润的胃癌病人，应尤为重视其MDR1mRNA表达的检测，以选择最适宜的化疗方案，达到满意的化疗目的。     

拓扑异构酶ⅡαRNA小分子干扰片段表达载体的构建与鉴定

目的：构建拓扑异构酶ⅡαRNA小分子干扰片段（TopoⅡαsiRNA）的真核表达载体并进行表达鉴定。方法：（1）根据GeneBank表达序列标签（Esr）数据库设计4对TopoⅡa基因siRNA的小分子片段。（2）采用RT—PCR方法检测细胞中的To—poⅡα表达并确定受试细胞。（3）将最佳沉默siRNA片段克隆到psilencer4．1-CMV-neo载体上,质粒DNA直接测序确定克隆成功。（4）采用脂质体法将TopoⅡa—siRNA DNA转染至受试细胞。并采用有限稀释法和G418加压筛选并培养稳转及对照细胞。（5）分别采用RT—PCR和Western blot方法测定稳转及对照细胞系TopoⅡαmRNA和蛋白表达。结果：（1）SKOV3／DDP细胞中TopoⅡαmRNA表达最高,与其亲本细胞SKOV3相比较,差异有统计学意义（P〈0．05）。（2）以TopoⅡα-4302siRNA片段沉默TopoⅡαmRNA表达最强（P〈0．01）。（3）重组阳性克隆经测序鉴定证实TopoⅡa-4302siRNA克隆成功。（4）RT—PCR和Western blot检测结果显示转染TopoIIa-4302siRNA质粒DNA后能抑制SKOV3／DDP细胞中的TopoⅡα基因表达。结论：成功构建psilencer4．1-CMV-neo—TopⅡα—siRNA重组表达载体并稳转至SKOV3／DDP细胞系。为下一步探讨TopoⅡα基因在卵巢癌多药耐药中的作用提供了实验基础。     

多药耐药相关蛋白和肺耐药蛋白在胃癌中的表达

目的：检测多药耐药相关蛋白（MRP）和肺耐药蛋白（LRP）在胃癌组织中的表达，并探讨其与临床病理特征的关系。方法：采用免疫组织化学法，检测52例患者胃癌组织和30例慢性胃炎组织中MRP及LRP的表达。结果：在52例胃癌组织和30例胃炎组织中，MRP的阳性表达率分别为32.69%（17例）、10％（3例），LRP的阳性表达率分别为63.46%（33例）、33.33%（10例），差异均有显著性意义（P＜0.05）。MRP的表达与胃癌病理分期和分化程度相关（P＜0.05）；LRP的表达与淋巴结转移相关（P＜0.05）。两蛋白在胃癌中的表达呈正相关（P＜0.05）。结论：MRP和LRP在胃癌组织中均有较高的表达，两者均可能是胃癌原发性多药耐药（MDR）的一种重要机制，且两者可能具有协同作用；临床上检测该两蛋白的表达，有利于为胃癌患者制定更加合理的综合治疗方案。     

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.