低浓度的丝裂霉素对结肠癌光动力增效作用研究

目的用低浓度的丝裂霉素改变大肠癌细胞周期来达到增强其光动力学疗法,探讨增效作用中对癌细胞杀伤作用规律。方法在结肠癌细胞培养的基础上,加入低浓度的丝裂霉素,光动力学疗法后采用荧光光度法和MTT法对癌细胞内光敏剂含量和细胞存活率进行检测,用SPLM软件统计包进行数据处理。结果低浓度的丝裂霉素可使癌细胞内光敏剂显著增加,随时间延长细胞内光敏含量增加明星;细胞内光敏剂含量与激光照射后细胞存活率呈显著负相关,即细胞内光敏剂含量多,PDT后细胞存活率低;光动力学疗法后癌细胞存活率曲线明显左移,在4、8、16h细胞存活率显著降低;癌细胞与光敏剂作用8～16h激光照射可达到最佳的光动力杀伤效果。结论我们设计单独低浓度的丝裂霉素对结肠癌细胞无杀伤作用,但可通过增加癌细胞内光敏剂含量来达到增效作用,8～16h为最佳给药时间,杀伤作用有明显的规律性。     

细胞外信号调节激酶通路激活抑制δ氨基酮戊酸-光动力疗法对SW480细胞的细胞毒作用

目的：探讨δ氨基酮戊酸-光动力疗法（ALA-PDT）诱导SW480细胞后的细胞外信号调节激酶（ERK）通路以及阻断此通路对细胞毒作用的影响。方法：将SW480细胞分为空白对照组、激光照射组、ALA组、ALA-PDT组（又分为30、60、90min组）及PD98059组,Western蛋白印迹检测各组细胞MEK和ERK1／2蛋白产物及磷酸化蛋白产物的表达;用MTT比色法分别检测各组细胞在不同时间段的光密度值,计算各组细胞存活率。结果：ALA-PDT后SW480细胞的ERK通路激活;阻断ERK通路后ALA-PDT对SW480细胞的细胞毒作用增强。结论：ERK通路的激活对ALA-PDT后SW480细胞起着保护作用;不同水平阻断ERK通路,可能成为增强ALA-PDT杀伤结肠癌细胞的新靶点。     

四溴苯三唑对人结肠癌SW480细胞凋亡的诱导作用及其机制

目的：探讨4,5,6,7-四溴苯三唑（TBB）对人结肠癌SW480细胞的促凋亡作用,并初步探讨其可能的作用机制。方法：选取处于对数生长期的人结肠癌SW480细胞,将其分为对照组（给予0 μmol·L^-1TBB）和实验组（给予1、3、10、30和100 μmol·L^-1TBB）。采用MTT比色法检测SW480细胞存活率,采用Annexin Ⅴ-FITC/PI双染法及流式细胞术检测SW480细胞凋亡率及细胞中活性氧（ROS）水平,采用Western blotting法检测细胞中抗凋亡蛋白p-Akt、Bcl-2和促凋亡蛋白Bad、pro-caspase-9及cleaved-caspase-3表达水平。结果：MTT法检测,与对照组比较,实验组SW480细胞存活率明显降低（P<0.05）,且呈浓度依赖性;Annexin Ⅴ-FITC/PI双染法及流式细胞术检测,随着处理时间的增加,与对照组比较,TBB作用3、6、12和24 h时SW480细胞凋亡率明显高于作用0 h时（P<0.05）;流式细胞术检测,TBB作用3、6、12和24 h时结肠癌SW480细胞中ROS水平高于作用0 h时（P<0.05）;Western blotting法检测,SW480细胞中抗凋亡蛋白p-Akt和Bcl-2表达水平逐渐降低,促凋亡蛋白Bad和cleaved-caspase-3表达水平逐渐升高,pro-caspase-9蛋白表达水平逐渐降低,与对照组（0 h）比较差异有统计学意义（P<0.05）。结论：TBB对体外培养的人结肠癌SW480细胞有明显的杀伤作用,可诱导细胞凋亡,其机制可能与抑制Akt活性或促进细胞中ROS产生有关。     

TRAIL联合奥沙利铂对结肠癌细胞株SW480凋亡的影响

目的 探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)联合奥沙利铂(oxaliplatin)对结肠癌细胞株SW480凋亡的影响,并初步探讨其作用机制.方法 应用MTT法分别检测TRAIL组、奥沙利铂组及联合组的细胞抑制率,Annexin Ⅴ-FITC/PI染色,流式细胞仪检测SW480细胞的凋亡率.结果 SW480细胞对TRAIL不敏感,对奥沙利铂相对敏感,TRAIL与亚毒性浓度的奥沙利铂联用对细胞的杀伤作用显著增强;亚毒性浓度的TRAIL、奥沙利铂及联合组作用SW480细胞24h的抑制率分别为8.62%,60.51%,81.28%;凋亡率分别为2.43%,11.76%,18.37%,流式细胞学证实这种杀伤作用主要通过诱导细胞凋亡实现.结论 TRAIL能显著提高奥沙利铂对结肠癌细胞的杀伤作用,此协同作用主要是通过诱导肿瘤细胞凋亡来实现.     

5-氨基酮戊酸-光动力疗法对HaCaT细胞增殖活性、细胞凋亡、原卟啉Ⅸ及活性氧的影响

目的观察5-氨基酮戊酸-光动力疗法(ALA-PDT)对Ha Ca T细胞增殖活性、细胞凋亡、原卟啉IX(Pp IX)及活性氧(ROS)的影响。方法体外培养Ha Ca T细胞,分成4组,对照组不加任何刺激因素,ALA组单用1 mmol·L-1ALA溶液加入Ha Ca T细胞培养基中避光孵育3 h,PDT组单用红光治疗仪照射Ha Ca T细胞培养液30 min,ALA-PDT组在加入1 mmol·L-1ALA溶液于Ha Ca T细胞培养基中避光孵育3 h后,用红光治疗仪照射上述培养液30 min。噻唑蓝(MTT)法检测细胞增殖活性,流式细胞仪检测细胞凋亡水平,荧光显微镜观察Pp IX的含量以及ROS检测试剂盒检测细胞内ROS的水平。结果对照组、ALA组、PDT组、ALA-PDT组细胞增殖活性分别为0.892±0.009、0.879±0.024、0.722±0.013、0.534±0.029;细胞凋亡率分别为(2.702±0.019)%、(2.697±0.031)%、(6.283±0.282)%、(32.601±1.897)%;ROS活性分别为(100.000±0.000)%、(101.333±3.786)%、(112.000±3.606)%、(135.000±16.000)%;PDT组细胞增殖活性、细胞凋亡率及ROS活性与对照组和ALA组比较差异有统计学意义(P<0.05);ALA-PDT组该3个指标与对照组、ALA组和PDT组比较,差异均有统计学意义(P<0.05)。对照组和ALA组细胞内Pp IX的荧光比率分别为(4.000±1.000)%、(32.000±2.646)%,2组比较差异有统计学意义(P<0.05)。结论 ALA诱导Ha Ca T细胞内Pp IX的累积,Pp IX在PDT作用下产生大量ROS,继而抑制Ha Ca T细胞增殖,导致细胞凋亡。     

辣椒素对人结肠癌SW480细胞株增殖作用的影响

目的 了解辣椒素对人结肠癌 SW480 细胞株增殖作用的影响,并探讨其可能的作用机制.方法 采用MTT检测法,观察终浓度100～400 μmol/L的辣椒素作用24、48 h对人结肠癌SW480 细胞株生长作用的影响;采用流式细胞仪检测400 μmol/L辣椒素作用人结肠癌 SW480 细胞株后其细胞周期变化和凋亡率.结果 辣椒素能抑制结肠癌 SW480 细胞株增殖,呈浓度、时间依赖性.在400μmol/L辣椒素作用下,结肠癌SW480细胞株停滞于G0/G1期.辣椒素作用24、48 h,SW480 细胞凋亡率升高(F=8.73、8.10,q=4.86～7.30,P<0.01).结论 辣椒素抑制 SW480 细胞增殖,并诱导其凋亡,其机制可能是阻滞细胞周期于G0/G1,期.     

羽扇豆醇对人共刺激细胞及结肠癌细胞株 SW480的影响

目的：共刺激细胞是一种对肿瘤细胞有杀伤作用的 NK样T细胞,既往研究表明羽扇豆醇作为一种天然植物提取物,能改变NK细胞、γδT细胞的生长及其对肿瘤细胞的作用。文中主要探讨羽扇豆醇对人共刺激细胞杀伤结肠癌细胞株 SW480的影响。方法取健康人外周血单个核细胞在体外经多种细胞因子诱导为共刺激细胞;不同浓度的羽扇豆醇在作用于共刺激细胞及结肠癌细胞株不同时间段后,甲基偶唑蓝（ MTT）法检测羽扇豆醇对共刺激细胞及结肠癌细胞株 SW480生长的影响;乳酸脱氢酶（ LDH）法检测共刺激细胞对结肠癌细胞株SW480的杀伤活性。结果羽扇豆醇浓度在0．1～200．0μg/mL时对共刺激细胞的生长有促进作用,对结肠癌细胞株SW480有抑制作用;羽扇豆醇诱导后,共刺激细胞对SW480的杀伤活性增强,浓度为12．5 mg/L时与空白对照比较的差异有统计学意义（76％vs 40％, P＜0．05）。结论羽扇豆醇能促进共刺激细胞的增殖,抑制结肠癌细胞株SW480的生长,并能增加SW480对共刺激细胞的敏感性,增强共刺激细胞对SW480细胞株的杀伤能力。     

青藤碱对人结肠癌细胞株SW480增殖和细胞周期的影响

目的 研究青藤碱对人结肠癌SW480细胞增殖和细胞周期的影响.方法 用不同浓度的青藤碱处理体外培养的人结肠癌SW480细胞24、48、72h后,采用CCK法检测细胞的增殖活性,采用流式细胞术检测细胞的细胞周期分布,并用光镜和透射电镜观察SW480细胞的形态学变化.结果 青藤碱在体外可抑制SW480细胞的增殖,并呈时间和剂量依赖性.干预48h后,中、低浓度的青藤碱（8mmol/L、4mmol/L）可诱导SW480细胞G1期细胞比例增加,而高浓度的SIN（16mmol/L、10mmol/L）可诱导G1期、G2期细胞比例增加.光镜与电镜下均见凋亡细胞增多.结论 青藤碱在体外能抑制人结肠癌SW480细胞增殖和诱导该细胞系的凋亡,其机制可能与其阻滞细胞周期进程有关.     

洛铂对结肠癌细胞的抗肿瘤活性及机制研究

目的探索洛铂在结肠癌细胞中的抗肿瘤活性,阐明潜在分子机制,为临床应用提供理论证据。方法体外培养结肠癌细胞SW480,用MTT比色法和克隆形成实验检测不同浓度的洛铂分别处理结肠癌SW480细胞株24、48 h和72 h后,对结肠癌细胞增殖活性的抑制作用;用流式细胞仪检测不同浓度梯度的洛铂对结肠癌SW480细胞株凋亡率的影响;采用Western blot检测洛铂对SW480细胞凋亡相关蛋白CASPASE-3、BAD、BCL-2的表达;添加AKT抑制剂及激动剂,采用Western blot检测CASPASE3、AKT、p AKT、BAD、p-BAD和BCL-2表达量,探讨洛铂促结肠癌细胞凋亡的作用机制。结果洛铂能显著抑制体外培养的SW480细胞的生长,呈现为时间剂量依赖型;洛铂可诱导结肠癌SW480细胞凋亡,且随着洛铂浓度的增加,结肠癌SW480细胞凋亡率显著提高,结果具有统计学意义。洛铂可下调结肠癌细胞抑凋亡基因BCL-2蛋白表达,上调促凋亡基因BAD和CASPASE-3蛋白表达量;添加AKT抑制剂MK-2206后,pAKT、p-BAD和BCL-2蛋白表达显著降低,CASPASE3蛋白表达显著升高,促进了细胞凋亡,结果具有统计学差异。添加AKT激动剂SC97处理后,pAKT、p-BAD和BCL-2蛋白表达显著上调,CASPASE3蛋白表达显著下调,抑制了细胞凋亡,结果具有统计学意义。结论洛铂能显著抑制人结肠癌SW480细胞增殖,诱导癌细胞凋亡,洛铂可能通过AKT/BCL-2/BAD信号通路发挥促细胞凋亡作用,是一种有效的抗结肠癌药物。     

PDT对SW480细胞和转染CD自杀基因的SW480结肠癌周期和凋亡的影响

采用脂质体基因转移技术将胞嘧啶脱氢酶（cytosinedeaminase,CD）自杀基因转染SW480结肠癌细胞后,实验分组为①空白对照组、②SW480组、③SW480-CD组,用半导体激光仪（距光斑3处输出功率12mW,能量密度9J／cm^2）垂直照射,照射8min后,用流式细胞仪分析8-氨基酮戊酸（8-aminolevulinic acid,ALA）—光动力学疗法（photodynamic therapy,PDT）结合CD／5Fc自杀基因系统对结肠癌细胞周期和凋亡的影响。     

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.