顺铂对人舌癌细胞的抑制作用和对端粒酶活性的影响

目的 研究化疗药物顺铂对人舌癌Tca8113细胞的抑制作用和对端粒酶活性的影响及其抗癌机制、并初步探讨端粒酶作为化疗敏感性指标的可行性。方法 应用顺铂处理的人舌癌Tca8113细胞为用药组，相同条件培养但不用药的细胞为对照组。MTT法研究细胞的生长和增殖情况，双层琼脂培养法研究细胞的克隆形成率，透射电镜观察细胞的超微结构，应用流式细胞术检测细胞周期，TRAP-PCR—ELISA法检测人舌癌Tca8113细胞端粒酶活性。结果 顺铂对人舌癌Tca8113细胞有明显的抑制作用，且呈浓度依赖性；能明显改变细胞大体形态和超微结构；抑制端粒酶活性且有一定的时间依赖性；细胞周期改变不明显。结论 顺铂可以明显抑制Tca8113细胞的增殖活性及其转移复发倾向、还可作用于线粒体，抑制端粒酶活性，对细胞周期的改变不明显。     

羟基喜树碱对人舌癌细胞的抑制作用及端粒酶活性的影响

目的研究化疗药物羟基喜树碱对人舌癌Tca8113细胞的抑制作用和对端粒酶活性的影响,初步探讨端粒酶作为化疗敏感性指标的可行性.方法应用MTT法、双层琼脂培养法、透射电镜观察、流式细胞术,TRAP-PCR-ELISA法研究.结果羟基喜树碱对人舌癌Tca8113细胞有明显的抑制作用且呈浓度依赖性,细胞超微结构有明显改变,细胞周期发生明显变化(S期细胞由23.9%变为39.5%,G0/G1期细胞由72.4%变为39.0%),抑制端粒酶活性且有一定的时间依赖性(用109ng/ml羟基喜树碱处理Tca8113舌癌细胞后24,48,72和96 h端粒酶活性分别为(0.57±0.07),(0.35±0.02),(0.18±0.04),(阴性).结论羟基喜树碱可以明显抑制Tca8113细胞的增值活性及其转移复发倾向,还可作用于线粒体,在细胞周期改变的同时抑制端粒酶活性.     

高三尖杉酯碱诱导白血病细胞凋亡与人端粒酶逆转录酶及端粒酶的关系

目的 研究高三尖杉酯碱(HHT)诱导白血病细胞调亡与细胞端粒酶和端粒酶逆转录酶(hTERT)水平的关系。方法 在体外，以HL60细胞株细胞为对象，采用半定量RT—PCR法检测hTERT mRNA表达量；采用端粒重复序列扩增法(TRAP)—酶联免疫吸附试验(ELISA)检测端粒酶活性；采用TUNEL法检测凋亡细胞。结果 与空白对照相比，HHT能够下调hTERT mRNA的表达。HHT浓度为0．005—0．030μg／m1时，HL60细胞hTERT mRNA表达无明显变化；当浓度增至0．040—0．050μg／m1，hTERT mRNA表达量下降至检测不出。0．020μg／m1 HHT作用18h后，hTERT mRNA表达呈下降趋势，24h后端粒酶活性明显下降，两者的下降趋势基本相似，但hTERT mRNA表达下降幅度较端粒酶活性下降幅度更大。在HHT作用下，HL60细胞凋亡率增加，而端粒酶活性及hTERT mRNA水平下降，且与HHT浓度或作用时间相关。结论 HHT下调端粒酶活性可能与hTERT mRNA转录水平下降有关。通过下降hTERT mRNA转录和端粒酶活性可能是HHT诱导HL60细胞凋亡的原因之一。     

siRNA特异性抑制舌癌细胞株端粒逆转录酶基因表达的体外研究

目的 探讨hTERT特异性的siRNA干扰舌癌Tca8113细胞hTERT基因表达的效果,和对舌癌Tca8113细胞增殖、凋亡的影响。方法 构建针对hTERT基因的siRNA（siRNA-hTERT1),转导入Tca8113细胞,同时以无同源性的siRNA-hTERT2作阴性对照,以及无任何处理的空白对照。应用实时聚合酶链反应(real-time PCR)技术检测hTERT基因表达变化,MTT法检测细胞生长率,流式细胞术分析细胞凋亡的情况。结果 与阴性对照组的1.934±0.212和空白对照的2.053±0.369相比,siRNA-hTERT1有效下调hTERT的mRNA表达仅为0.950±0.138,P＜0.01。转染72 h后,siRNA-hTERT1的细胞抑制率达47.2％,远远高于阴性对照组的2.6％,P＜0.01。流式细胞仪检测结果显示,siRNA-hTERT1处理48h后细胞凋亡率明显增加,达(27.30±0.18)％,显著高于阴性对照组的(5.11+0.22)%和空白对照组的(5.00+0.19)%,P＜0.01。结论 siRNA-hTERT1能特异性抑制舌癌Tca8113细胞hTERT基因的表达,促进细胞的凋亡和减慢细胞增殖。     

化疗药物诱导肝癌细胞凋亡过程中端粒酶、端粒酶逆转录酶活性的变化

目的:探讨hTERT和端粒酶在化疗药物诱导凋亡过程中的作用及其意义。方法选择IC50剂量的三尖杉酯碱、长春新碱和足叶已甙处理肝癌细胞,采用TRAP法检测药物处理前后肝癌细胞端粒酶活性;应用流式细胞学检测细胞周期和细胞凋亡率;并利用Western Blotting检测药物作用后hTERT蛋白水平变化。结果三尖杉酯碱、长春新碱作用于肝癌细胞,明显降低hTERT蛋白表达水平,下调胞内端粒酶活性,并诱导细胞凋亡及周期阻滞;足叶已甙对肝癌细胞端粒酶活性及hTERT蛋白水平无明显抑制作用;化疗前后胞内hTERT水平和细胞凋亡率的变化呈明显负相关。结论化疗药物可能通过降低hTERT蛋白水平而下调了端粒酶活性,并诱导凋亡;hTERT蛋白水平可作为化疗敏感性指标。     

蟾毒灵体外抑制人舌鳞癌Tca8113细胞增殖和促凋亡作用及对Na+/K+-ATP酶的影响

目的 探索蟾毒灵对舌鳞状细胞癌Tca8113细胞增殖、凋亡的影响及可能作用机制.方法 以人舌鳞癌Tca8113细胞为研究对象,MTT法检测10、20、40、80、160 nmol/L浓度蟾毒灵体外抑制Tca8113细胞增殖的活性;检测蟾毒灵干预下肿瘤细胞Na+-K+-ATP酶活性的变化;Western blot发检测Bcl-2、Bax、caspase-3蛋白表达.结果 蟾毒灵有抑制Tca8113细胞的活性,且呈剂量-时间依赖性;在蟾毒灵干预下Tca8113细胞Na+-K+-ATP酶收到抑制;Western blot结果显示凋亡相关Bax、caspase-3蛋白表达上调,Bcl-2蛋白表达下调.结论 蟾毒灵通过抑制细胞膜Na+-K+-ATP酶活性,通过调节Bcl-2凋亡通路的相关蛋白,最终激活caspase-3,诱导人舌鳞癌Tca8113细胞凋亡.     

hTERT反义寡核苷酸对乳腺癌细胞增殖的影响

目的探讨hTERT反义寡核苷酸抑制端粒酶活性对乳腺癌细胞增殖的影响。方法用脂质体介导反义寡核苷酸转染MCF-7乳腺癌细胞株,采用TRAP-ELISA法检测转染后细胞端粒酶的活性,观察其前后的变化。MTT法检测细胞活性,流式细胞仪观察细胞周期及细胞凋亡率变化。结果转染细胞的端粒酶活性及细胞生长都明显受到抑制。流式细胞仪检测到细胞凋亡峰,细胞阻滞于S期,正义寡核苷酸则无此作用。结论端粒酶hTERT为靶点的反义寡核苷酸可明显抑制乳腺癌细胞的增殖,且具有促凋亡作用。     

阿霉素诱导Tca8113细胞凋亡及其分子机制的研究

【目的】观察阿霉素诱导Tca8113细胞凋亡及其过程中某些凋亡相关分子表达的变化,探讨其诱导凋亡作用的分子机制。【方法】采用MTT法检测阿霉素对Tca8113细胞的增殖作用;以光镜、荧光纤维镜、流式细胞仪和Annexin V-FITC标记法检测细胞凋亡;以蛋白印迹法分析阿霉素对Bcl-2、Bax和Survivin蛋白表达的影响。【结果】阿霉素以剂量依赖方式抑制Tca8113细胞增殖;光镜及荧光显微镜下可见明显凋亡细胞;流式细胞仪检测显示细胞停滞于G1或S期,有明显凋亡峰出现;Annexin V-FITC标记法定量检测证实阿霉素可诱导细胞凋亡发生;蛋白印迹检测表明阿霉素可使Bax表达升高,Bcl-2和Survivin表达下降。【结论】阿霉素可显著抑制Tca8113细胞增殖,可通过调节Bcl-2、Bax和Survivin蛋白表达来实现致凋亡作用。     

miR-200c对舌癌Tca8113细胞增殖和凋亡的影响

目的：探讨 miR-200c 对人舌鳞状细胞癌 Tca8113细胞增殖和凋亡的影响。方法将 miR-200c 的模拟物通过脂质体转染到人舌鳞状细胞癌 Tca8113细胞内,通过四甲基偶氮唑盐（MTT）法检测细胞增殖能力、流式细胞术检测细胞周期和细胞凋亡率、Western blot 检测半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）和 Bcl-2蛋白的表达。结果与对照组相比,miR-200c 模拟物20、40、80 nmol／L 组的 Tca8113细胞生长受到明显抑制,差异有统计学意义（P ＜0．05）,miR-200c 模拟物的浓度越大,作用时间越长,抑制效果越明显（P ＜0．05）;转染 miR-200c 模拟物48 h 后,Tca8113细胞停滞于 G0／G1期,细胞凋亡率显著增加,差异有统计学意义（P ＜0．05）;Western blot 证明20、40、80 nmol／L 组 Bcl-2蛋白表达量明显低于对照组,而 Caspase-3蛋白表达量明显高于对照组（P ＜0．05）。结论miR-200c 过表达可抑制舌癌 Tca8113细胞增殖并诱导细胞凋亡。     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

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.