氯硝柳胺逆转KBv200细胞对阿霉素耐药性的机制研究

目的:探讨氯硝柳胺对人口腔表皮癌多药耐药细胞株KBv200耐药的逆转作用及其机制。方法:采用MTT法评价氯硝柳胺对KBv200细胞药物敏感性的影响,流式细胞术检测氯硝柳胺对细胞内化疗药物阿霉素积累的影响,Annexin V-FITC/PI双染法测定细胞凋亡情况,Western blot分析氯硝柳胺对多药耐药蛋白ABCB1表达的调控。结果:无毒剂量的氯硝柳胺(5μmol/L)增加KBv200细胞对阿霉素的敏感性,并显著提高阿霉素在细胞中的积累。此外,氯硝柳胺联合阿霉素可促进细胞凋亡。氯硝柳胺作用后,多药耐药蛋白ABCB1的表达水平下降。结论:氯硝柳胺对KBv200细胞的阿霉素耐药性有逆转作用,其机制与下调ABCB1蛋白表达有关。     

洛美利嗪逆转K562/A02细胞多药耐药的研究

目的 :研究洛美利嗪对肿瘤多药耐药的逆转作用 ,探讨洛美利嗪逆转肿瘤细胞多药耐药的机制。方法 :以人红白血病细胞系K5 6 2及其耐药细胞系K5 6 2 /A0 2 (耐阿霉素 )为研究对象 ,采用MTT法检测细胞毒作用 ;免疫组化测定法检测P 糖蛋白 (P glycoprotein ,P gp)表达水平 ;RT PCR法检测mdr1mRNA水平 ;以流式细胞术测定两种细胞系内罗丹明 12 3(Rh12 3)的潴留来反映P gp的外排功能。结果 :与K5 6 2细胞系相比 ,K5 6 2 /A0 2耐药细胞系mdr1mRNA及P gp高表达 ,Rh12 3潴留减少。洛美利嗪 (10 μmol/L)对K5 6 2 /A0 2细胞的mdr1mRNA及P gp表达水平无明显影响 (P >0 .0 5 )。洛美利嗪对K5 6 2 /A0 2细胞的阿霉素 (ADM)、长春新碱 (VCR)细胞毒性有剂量相关性增敏作用。洛美利嗪能增加K5 6 2 /A0 2耐药细胞系的Rh12 3潴留。结论 :洛美利嗪对K5 6 2 /A0 2细胞的ADM、VCR细胞毒性有剂量相关性增敏作用 ,其机制可能与增加细胞内药物浓度有关。     

乙酰阿地咪逆转肿瘤细胞多药耐药及其机制的研究

目的 探讨乙酰阿地咪体外逆转多药耐药细胞株人乳腺癌耐阿霉素细胞（MCF-7/Adr）以及人非小细胞肺癌耐阿霉素细胞（A549/Adr）的耐药作用及其作用机理。方法 采用乳酸脱氢酶释放法检测乙酰阿地咪与阿霉素共处理MCF-7、A549、MCF-7/Adr以及A549/Adr细胞的细胞毒作用,以观察增敏和逆转耐药作用;用全功能酶标仪测定乙酰阿地咪与阿霉素共同处理MCF-7/Adr和A549/Adr细胞后细胞内积累阿霉素的荧光强度,并采用Western blot检测细胞膜上多药耐药蛋白P-糖蛋白（P-gp）表达的变化。结果 乙酰阿地咪能降低MCF-7/Adr和A549/Adr对阿霉素的耐药性,10 μmol/L的乙酰阿地咪对MCF-7/Adr和A549/Adr逆转倍数分别为10.8、20.1,其逆转作用与乙酰阿地咪呈剂量依赖关系,且对MCF-7和A549细胞也具有增敏作用;随着乙酰阿地咪浓度的增加,MCF-7/Adr和A549/Adr细胞内积累阿霉素的荧光强度增加率增加,呈剂量依赖性;经乙酰阿地咪处理的MCF-7/Adr和A549/Adr细胞P-gp蛋白表达水平降低,且降低水平随乙酰阿地咪浓度增加而更明显。结论 乙酰阿地咪可显著逆转肿瘤细胞的多药耐药性,其机理与抑制多药耐药蛋白P-gp的表达有关,提示乙酰阿地咪具有潜在的克服肿瘤化疗中多药耐药现象的应用价值。     

热化疗联合维拉帕米对人乳腺癌耐药细胞化疗敏感性的实验研究

目的观察用热化疗与维拉帕米合用是否能抑制人乳腺癌耐药细胞株MCF-7/ADM多药耐药蛋白P-gp的表达以逆转乳腺癌对化疗药的耐药性,提高化疗敏感性。方法高表达P-gp的MCF-7/ADM,在37~43℃热化疗条件下分别加入3μg/mL阿霉素(ADM)和不同浓度的逆转剂维拉帕米(VRP,0~5μmol/L),用共聚焦MRC-1024(60×物镜)观察扫描,Laserpix软件检测细胞内ADM的荧光强度,流式细胞仪检测凋亡。结果随着温度和VRP浓度的升高,MCF-7/ADM细胞内ADM荧光强度升高,凋亡率增加,且温度和浓度有协同作用。结论热化疗与维拉帕米联合可有效逆转P-gp所致耐药,提高化疗敏感性。     

小分子干扰RNA片段逆转KBv200细胞多药耐药的实验研究

目的：研究小分子干扰RNA片段（siRNA）对舌癌多药耐药（MDR）细胞系KBv200细胞的MDR1基因表达及其功能的影响，探讨siRNA作为未来化疗增敏剂的可能性。方法：运用针对MDR1基因序列的siRNA（MDR1-siRNA）在脂质体的介导下转染KBv200细胞；用RT—PCR分析MDR1 mRNA的表达水平；流式细胞术检测P-糖蛋白（P-gP）的表达；荧光分光光度法检测细胞内多柔比星（Dox）的积蓄；MTT法检测KBv200细胞对长春新碱（VCR）和Dox的敏感性变化。结果：MDR1-siRNA作用24和48h能抑制MDR1基因的表达（P〈0．05），其mRNA水平和P-gP水平均明显下降，同时使KBv200细胞对VCR和Dox的敏感性显著增加（P〈0．01），并显著增加细胞内Dox的蓄积（P〈0．01）。结论：MDR1-siRNA能显著抑制KBv200细胞内MDR1基因介导的MDR。     

改良微量荧光分光光谱法测定卵巢癌细胞内阿霉素聚集量

目的 :测量卵巢癌SKOV3 亲本和耐药细胞内阿霉素聚集量。方法 :采用一种简单敏感的改良荧光分光光谱法。结果 :虽然耐药细胞株SKOV3/ADM内阿霉素聚集量经维拉帕米作用后能显著增加 ,但仍然低于亲本细胞SKOV3。结论 :本法快速、方便 ,在肿瘤细胞多药耐药机理及逆转剂研究中具有实用价值。     

粉防己碱对喉癌耐药细胞株KBV200耐药逆转作用的研究

目的:研究天然有效成分粉防己碱(Tetrandnne,TET)对喉癌耐药细胞株KBV200的耐药逆转作用.方法:本研究以喉癌耐药细胞株KBV200为研究对象,采用粉防己碱逆转KBV200对常用抗肿瘤药物阿霉素的多药耐药作用,以经典逆转剂维拉帕米(Verapamil,VRP)为对照组,用四甲基偶氮唑蓝(MTT)法测定其对实验细胞的增殖抑制率.结果:低毒剂量的粉防己碱能逆转实验细胞株埘阿霉素的多药耐药(muhidrug resistance,MDR)作用,逆转指数为22.96;维拉帕米逆转指数为9.31.结论:低毒浓度的天然药物粉防己碱能逆转喉癌耐药细胞株KBV200对阿霉素的耐药作用,其逆转效果优于经典逆转剂维拉帕米.     

丹参对人胃癌阿霉素耐药性的逆转作用

[目的]探讨丹参对人胃癌多药耐药的逆转作用。[方法]以不同浓度的丹参处理胃癌SGC7901及耐药SGC7901/ADR细胞,MTT法检测丹参对两种细胞的毒性及半数抑制率（IC50）,流式细胞仪（FCM）检测两种细胞内阿霉素浓度和细胞周期。胃腺癌耐药患者24例,随机分为二组,术前分别用丹参12 g.d^-1与阿霉素10mg.m^-2和单用阿霉素10 mg.m^-2,术后光镜、电镜观察癌组织变化和高效液相色谱法（HPLC）测定细胞内阿霉素浓度。[结果]丹参浓度在500μg/mL以下无细胞毒性,使SGC7901/ADR细胞对阿霉素的IC50由6.01 mg/L增为3.17 mg/L,耐药性得到部分逆转,逆转倍数为1.89（P〈0.01）;细胞内阿霉素浓度由1.83增加到2.83,增加1.55倍（P〈0.05）;使细胞周期阻滞于G1、G2期;体内丹参12 g.d^-1与阿霉素合用时癌细胞损伤加重,线粒体肿胀嵴变短,数目减少,严重时内、外膜破裂,内质网扩张和囊泡化,伴有膜旁核蛋白体的脱落;癌细胞坏死增多;细胞内阿霉素浓度由45.76增加到133.07,增加2.90倍。[结论]丹参对SGC7901/ADR细胞和胃腺癌阿霉素多药耐药有逆转作用,逆转机制与丹参能增加耐药细胞内化疗药物浓度有关。     

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。     

中药活性成分逆转肿瘤细胞多药耐药作用的体外筛选

①目的 从多种中药单体或提取物中筛选肿瘤细胞多药耐药(MDR)逆转剂。②方法 采用MTT法进行药物细胞毒实验及筛选与逆转作用研究。③结果 黄芩甙、甘糖酯、茶多酚、苦参碱，在已出现明显细胞毒性的剂量下，仍未显示对长春新碱(VCR)耐药的逆转作用。钩藤总碱5.0μg／ml，药根碱2μg／ml(5．9μM)，靛玉红1．25μg／ml(4．8μM)，姜黄素1．56fμg／ml(17μ)、对VCR耐药的逆转倍数分别为l6．8倍、5．1倍、4倍及1．97倍。而在亲本KB细胞，除了姜黄素外，其它3种药物对VCR细胞毒性无明显影响。④结论 钩藤总碱、药根碱、靛玉红对KBv200细胞耐药具有逆转作用，而姜黄素与VCR合用在KB及KBv200细胞均有增敏作用。     

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.