葡萄籽多酚对多药耐药的逆转作用及其机制

目的：探讨葡萄籽多酚(GSPs)逆转多药耐药的作用及其对多药耐药相关蛋白GSTπ、TopoⅡ α表达的影响。方法：以人乳腺癌MCF-7细胞及其耐药株MCF-7／ADR细胞作为实验对象，用MTT法观察GSPs对多药耐药的逆转作用，免疫细胞化学技术观察GSPs对GSTπ、TopoⅡα等多药耐药相关蛋白表达的影响。结果：GSPs在1．2、2．4mg／L浓度时对细胞无毒性作用，并且均能逆转MCF-7／ADR细胞的多药耐药性，逆转倍数分别为5．77和9．79倍；GSPs可使MCF-7／ADR细胞GSh表达下降，但对TopoⅡ α表达无影响。结论：体外试验证实GSPs具有多药耐药逆转作用，其机制可能与降低肿瘤细胞内GSTπ的表达有关，提示GSPs是一种潜在的肿瘤化疗增敏剂。     

Alopecurone B逆转人乳腺癌细胞MCF-7阿霉素耐药株活性研究

目的 研究黄酮化合物Alopecurone B（APB）对人乳腺癌细胞阿霉素耐药株MCF-7/ADR多药耐药的逆转活性及逆转机制。方法 人乳腺癌细胞阿霉素耐药株MCF-7/ADR维持在含有250 ng/mL阿霉素的培养基中,使用MTT法、qPCR、Western blot和流式细胞术研究APB对耐药株P-糖蛋白(P-gp)功能和表达的影响。结果 APB能逆转MCF-7/ADR细胞多药耐药,抑制P-gp的功能,下调P-gp基因和蛋白的表达。结论 APB具有强效逆转人乳腺癌细胞MCF-7/ADR多药耐药的效果,其机制涉及对P-gp的调控。      

TopoⅡ、GST-π、P-gp在乳腺癌的表达及意义

目的探讨几种多药耐药机制与乳腺浸润性导管癌的耐药性及临床病理特征的关系。方法应用免疫组化S-P法检测DNA拓扑异构酶Ⅱ、谷胱甘肽-S-转移酶-π和P-糖蛋白在220例乳腺浸润性导管癌组织中的表达。结果乳腺浸润性导管癌病例GST-π、P-gp均有较高的阳性率,TopoⅡ有近一半的病例不表达。TopoⅡ、GST-π和P-gp的表达与乳腺浸润性导管癌的分级、淋巴结转移无明显相关性(P>0.05)。结论乳腺浸润性导管癌的多药耐药与TopoⅡ、GST-π、P-gp的耐药机制有关。乳腺浸润性导管癌组织TopoⅡ、GST-π、P-gp的表达可作为指导乳腺浸润性导管癌化疗的重要指标和判断乳腺浸润性导管癌预后的参考指标。     

实时荧光定量PCR检测乳腺癌5种多药耐药基因的表达强度

目的 :检测 5种多药耐药基因在乳腺癌细胞株 MCF- 7和 MCF- 7/ADR里的表达强度变化 ,为乳腺癌多药耐药逆转研究提供新的思路。方法 :通过实时荧光定量 PCR技术分别检测乳腺癌敏感细胞株 MCF- 7和耐药细胞株 MCF- 7/ADR里的 mdr1、MRP、L RP、GST- π、TOPO α基因表达强度。结果 :MCF- 7/ADR里的 mdr1、MRP、LRP、GST- π基因表达强度分别比它们在 MCF- 7里的表达强度相对增加了 1 2 5 .39、5 5 .5 3、1 .2 4、3.38倍 ,TOPO α则降低了 0 .43倍 ,其中 mdr1、MRP表达强度的相对增加倍数均显著高于 L RP、GST- π、TOPO α的增加 (或降低 )倍数 ( P<0 .0 0 1 )。结论 :除了 mdr1之外 ,MRP也可能在乳腺癌的多药耐药产生机制里起重要作用。因此 MRP基因可以成为乳腺癌耐药逆转研究的新作用靶点     

胎盘型谷胱甘肽-S-转移酶和DNA拓朴酶在人骨肉瘤中的表达及其与预后的关系

目的探讨骨肉瘤组织中胎盘型谷胱甘肽-S-转移酶(GST-π)和DNA拓朴酶Ⅱ(Topo Ⅱ)的表达及其与预后的关系.方法应用免疫组化SP法检测GST-π、Topo Ⅱ在40例骨肉瘤中的表达,并结合临床资料分析其对预后的影响.结果 40例骨肉瘤中GST-π和Topo Ⅱ的阳性表达率分别为82.5%(33/40)和52.5%(21/40),其中术前化疗组Topo Ⅱ阳性表达率为38.5%(5/13),显著低于术前无化疗组77.8%(21/27)(P<0.05),GST-π的阳性表达率在两组间无显著差异;GST-π和Topo Ⅱ的阳性表达率与患者的预后显著相关.结论 GST-π主要参与骨肉瘤原发性耐药,Topo Ⅱ不仅参与骨肉瘤原发性耐药,同时也参与骨肉瘤继发性耐药;GST-π和Topo Ⅱ是影响骨肉瘤预后的重要因素,可能成为评估骨肉瘤预后的有价值的指标.     

非小细胞肺癌组织中Ki67、多药耐药基因GST-π/TopoⅡ/P-gp的表达及其与临床病理学相关性研究

目的探讨Ki67在非小细胞肺癌(non-small cell lung carcinoma,NSCLC)组织中的表达及其与多药耐药基因谷胱甘肽-S转移酶π(GST-π)、拓扑异构酶Ⅱ(TopoⅡ)、P-糖蛋白(P-gp)表达的相关性及其意义。方法采用免疫组化法检测108例NSCLC患者癌组织中Ki67、GST-π、TopoⅡ、P-gp的表达水平。结果 Ki67、GST-π、TopoⅡ、P-gp在108例NSCLC患者癌组织中的阳性表达率分别为95.37%、81.48%、82.40%、38.89%;GST-π表达与NSCLC分化程度有关(χ~2=6.318,P=0.042);Ki67表达与TopoⅡ表达有关(χ~2=6.503,P=0.011),共表达率为80.56%,但关系不密切(r=0.245,P=0.010);Ki67表达与GST-π表达有关(χ~2=4.686,P=0.03),共表达率为76.85%,但是相关性低(r=0.028,P=0.031);Ki67表达与P-gp表达无关(P>0.05)。GST-π与TopoⅡ表达呈正相关(r=0.028,P=0.023),共表达率为70.37%,但是关系不密切;P-gp与GST-π、TopoⅡ无相关(P>0.05)。结论 Ki67可能不参与NSCLC细胞的多药耐药机制;三种多药耐药基因可能是通过不同的途径介导NSCLC多药耐药。     

乳腺癌组织中P-gp、GST-π与TopoⅡ耐药基因的表达及临床意义

目的探讨多药耐药基因蛋白P-糖蛋白(P-gp)、谷胱甘肽-S-转移酶-π(GST-π)及DNA拓扑异构酶Ⅱ(TopoⅡ)在乳腺癌中的表达及其临床意义。方法用免疫组化法检测52例未经抗肿瘤治疗的原发乳腺癌中P-gp、GST-π及TopoⅡ的表达水平。结果52例乳腺癌组织中P-gp、GST-π和TopoⅡ表达率分别为57.7%、53.8%和76.9%,P-gp、GST-π和TopoⅡ的表达与组织学类型、年龄无关(P>0.05),与临床分期、淋巴结转移有显著性差异(P<0.05),P-gp和GST-π的表达呈显著正相关(P<0.05),P-gp和TopoⅡ的表达呈明显负相关(P<0.05),GST-π和TopoⅡ呈明显负相关(P<0.05)。结论乳腺癌化疗前检测P-gp、GST-π及TOPOⅡ耐药基因蛋白,对肿瘤的化疗药物选择及判断预后有一定的价值。     

钙调素拮抗剂O-(4-乙氧基)-丁基-小檗胺对乳腺癌多药耐药细胞系MCF-7/ADR的耐药逆转作用

目的研究钙调素拮抗剂O-4-乙氧基-丁基-小檗胺(EBB)对乳腺癌多药耐药细胞系MCF-7/ADR的耐药逆转作用。方法采用四唑盐(MTT)比色法测定药物敏感性,分析联合应用低剂量EBB(≤IC20)时阿霉素(ADR)对MCF-7/ADR细胞的半数抑制浓度(IC50)值及其增敏倍数;应用流式细胞术测定EBB对细胞周期的影响,同时观察细胞内药物浓度的积累;采用逆转录—多聚酶链反应检测EBB对多药耐药基因-1(mdr1)及拓扑异构酶Ⅱb(topⅡb)mRNA表达水平的影响。结果EBB对MCF-7/ADR的多药耐药具有明显逆转作用,3、7·5μmol/L浓度的EBB可以明显提高MCF-7/ADR对ADR的敏感性,平均增敏倍数分别为50·40、89·80倍,逆转强度明显高于阳性逆转剂维拉帕米(VPL)的14·88倍(P=0·0097)。6μmol/L的EBB处理2h后,细胞内积累的P糖蛋白底物罗丹明123(Rh123)荧光强度峰值发生明显右移;同时EBB可明显增强ADR对MCF-7/ADR细胞在G2/M期的阻滞作用。6和12μmol/L的EBB处理MCF-7/ADR细胞48h后,mdr1的mRNA表达水平有一定的降低趋势,但无统计学差异;topⅡb基因表达差异亦无显著性。结论EBB对MCF-7/ADR细胞具有较强的逆转多药耐药作用,其可能的逆转机制在于增强ADR对耐药细胞在G2/M期的阻滞以及抑制P糖蛋白外排泵作用。     

多药耐药基因产物和激素受体在乳腺癌中的表达及意义

目的：探讨多药耐药基因产物谷胱甘肽转移酶π（ GST-π）、肺耐药相关蛋白（ LRP）和DNA拓扑异构酶Ⅱ（ TopoⅡ）和雌激素受体（ ER）、孕激素受体（ PR）在乳腺浸润性导管癌组织中的表达及临床意义,并分析多药耐药基因产物与雌、孕激素受体之间是否有关联。方法应用免疫组织化学SP二步法检测194例乳腺浸润性导管癌组织中GST-π、LRP、TopoⅡ、ER、PR的表达,且结合乳腺癌临床病理因素进行分析。结果194例乳腺浸润性导管癌组织中GST-π、LRP、TopoⅡ表达的阳性率分别为62.4％、81.4％、51.5％。 LRP和TopoⅡ在伴有淋巴结转移的乳腺浸润性导管癌患者中的表达高于无淋巴结转移者（ P＜0.05）,而与患者年龄、肿块长径、组织学分级、TNM分期等临床病理因素之间无相关性（P＞0.05）;LRP和TopoⅡ的表达与ER、PR的表达均无明显相关性（ P＞0.05）。 GST-π的表达与ER、PR的表达呈负相关（ P＜0.05）,而与其它临床病理因素之间未见相关性（ P＞0.05）。 LRP、GST-π、TopoⅡ两两之间无相关性（ P＞0.05）。结论 GST-π、LRP、TopoⅡ与乳腺浸润性导管癌的多药耐药有关,且LRP和TopoⅡ与乳腺癌的转移有一定关系,GST-π与ER、PR负性相关。联合检测可为乳腺癌的临床化疗药物选择及评估预后提供依据。     

逆转胶囊含药血清对MCF-7／ADR细胞bcl-2基因表达的影响

目的探讨中药逆转胶囊对乳腺癌多药耐药细胞株MCF-7／ADR细胞bcl-2基因表达的影响从而进一步了解其逆转耐药的作用机制。方法用逆转胶囊含药血清培养MCF3／ADR细胞,同时设计实验分组,以流式细胞仪计数方法检测各组MCF-7／ADR细胞的bct-2基因的表述情况。结果逆转胶囊含药血清能显著抑制MCF-7／ADR细胞bcl-2基因的表达;与阿霉素合用时亦可抑制MCF-7／ADR细胞bcl-2基因的表达,加强ADM诱导凋亡的作用。结论逆转胶囊具有逆转MCF-7／ADR多药耐药性的作用,其机制之一可能是通过下调bcl—2基因表达来实现的。     

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.