Iressa与顺铂联合抑制舌鳞癌细胞增殖的实验研究

 【目的】探讨EGFR靶向治疗药物Iressa与顺铂联用对舌鳞癌细胞增殖的抑制作用。【方法】采用MTT的方法检测Iressa与顺铂联用对舌鳞癌细胞系Tscca和Tca8113抑制作用的效率。【结果】Iressa抑制Tscca和Tca8113细胞增殖的IC50值分别为18.03 μmol/L和29.98 μmol/L(P=0.002);Iressa与顺铂联用加强了对舌鳞癌细胞系Tscca、Tca8113的增殖抑制,而且当Iressa提前24h或48h使用时,这种协同作用更加明显。【结论】Iressa可抑制舌鳞癌细胞系Tscca和Tca8113的增殖,与顺铂联用可协同抑制Tscca和Tca8113的增殖,这种抑制作用具有浓度和顺序依赖性。     

舌鳞癌细胞对Iressa的敏感性及其表皮生长因子受体突变

目的研究中国人舌鳞癌细胞系对表皮生长因子受体(EGFR)抑制剂Iressa的敏感性及其EGFR突变,初步探讨Iressa在头颈鳞癌治疗中的价值。方法以中国人舌鳞癌细胞系Tscca和Tca8113为研究对象,采用MTT法检测Iressa对其增殖的抑制作用;流式细胞仪检测细胞周期和凋亡;对Tscca和Tca8113进行EGFR外显子18～21测序。结果 Iressa对Tscca、Tca8113均有显著的增殖抑制作用,呈剂量依赖性。细胞周期分析显示,较低浓度Iressa(18.2μmol/L和36.4μmol/L)可使Tscca和Tca8113细胞G0/G1期比例增高;流式细胞仪分析显示高浓度Iressa可诱导Tscca和Tca8113细胞凋亡。测序发现Tscca和Tca8113的EGFR外显子20存在同一静止突变,即2361G-A,Q787Q。结论 Iressa可抑制Tscca和Tca8113的增殖,呈剂量依赖性;较低浓度时诱导细胞周期G0/G1期比例增高,高浓度时诱导凋亡,其抑制作用与EGFR突变未见相关性。     

姜黄素对人口腔鳞癌细胞增殖及凋亡的影响

目的探讨姜黄素对人口腔鳞癌细胞Tca8113及Tb增殖的抑制及诱导凋亡作用。方法分别应用6．25、12．50、25．00、50．00μmol／L的姜黄素作用Tca8113及Tb细胞24h,采用MTT法检测姜黄素对口腔鳞癌细胞Tca8113及Tb增殖的抑制作用,形态学观察及流式细胞仪测定细胞凋亡。结果6．25～50．00μmol／L姜黄素能显著抑制口腔鳞癌细胞Tca8113和Tb细胞的增殖（F＝793．94、391．08,q＝6．00～62．68,P〈0．05）。流式细胞仪检测显示口腔鳞癌细胞凋亡率随姜黄素浓度升高而增加。倒置显微镜下可观察到部分凋亡细胞出现染色质凝集、核固缩、核碎裂等形态学改变。结论姜黄素对人口腔鳞癌细胞Tca8113及Tb的增殖具有显著的抑制作用,并可诱导细胞凋亡。     

姜黄素通过P53通路诱导人舌鳞癌细胞的凋亡

目的:检测姜黄素对舌鳞癌细胞Tca-8113增殖和凋亡的影响,研究姜黄素对p53信号通路的影响。方法:使用浓度为10、20、40、80 mol/L的姜黄素与舌鳞癌细胞系Tca-8113共培养,在24 h和48 h后采用MTT法检测细胞的增殖情况;使用20、40 mol/L姜黄素与Tca-8113共培养24 h后,采用Western-Blot检测p53、p21和caspase9蛋白的表达。结果:姜黄素能够显著抑制Tca-8113细胞的增殖,抑制作用呈时间依赖性和药物浓度依赖性;姜黄素能够促进p53、p21和caspase 9蛋白的表达。结论:姜黄素能够抑制Tca-8113细胞的增殖并促进其凋亡,这一机制可能和p53信号通路有关。     

JSH-23对人舌鳞癌细胞Tca8113增殖和凋亡的影响

目的 探讨JSH-23阻断NF-κB信号通路后人舌鳞癌细胞Tca8113增殖和凋亡的情况及NF-κB信号通路相关凋亡抑制基因Bcl-2的表达变化,进而为临床舌鳞癌的干预和治疗寻找新的药物提供实验依据.方法 采用体外培养人舌癌Tca8113细胞的方法,经不同浓度JSH-23作用不同时间后,用MTT法、RT-PCR及Western blot等方法检测JSH-23对Tca8113细胞增殖、凋亡的影响.采用SPSS13.0统计软件对数据进行方差分析及t检验.结果 Tca8113细胞经JSH-23作用后,增殖受到明显抑制,20 μmol/L JSH-23作用48 h时,抑制作用最为显著,细胞生长抑制率达到60.45％.同时,Bcl-2凋亡抑制基因表达减少,与对照组相比,JSH-23作用48 h后,Bcl-2表达显著减少47.85％,差异有统计学意义（P＜0.05）;Bcl-2蛋白含量也明显减少,作用48 h后,Bcl-2蛋白表达相对光密度值分别下降50.93％ （P ＜0.01）.结论 JSH-23作为NF-κB信号通路抑制剂,可以明显抑制人舌鳞癌细胞的增殖,同时显著下调Bcl-2基因及蛋白的表达.     

诱导型一氧化氮合酶和血管内皮生长因子在舌癌细胞株中的表达

 【目的】探讨诱导型一氧化氮合酶（iNOS）和血管内皮生长因子（VEGF）在舌鳞癌细胞株Tca8113中的表达,为进一步研究iNOS和VEGF在舌鳞癌肿瘤血管生成中的作用提供实验基础。【方法】采用逆转录-聚合酶链反应（RT-PCR）法和SP免疫组化方法,对舌鳞癌细胞株Tca8113中iNOS和VEGF mRNA和蛋白表达情况进行检测。【结果】RT-PCR检测到舌鳞癌Tca8113细胞株中iNOS／VEGF mRNA条带明显,SP免疫组化检测到iNOS／VEGF蛋白在舌癌细胞胞浆中呈强阳性表达。【结论】iNOS／VEGF mRNA和蛋白在舌鳞癌Tca8113细胞株中呈高水平表达。     

诱导型一氧化氮合酶和血管内皮生长因子在舌癌细胞株中的表达

【目的】探讨诱导型一氧化氮合酶（iNOS）和血管内皮生长因子（VEGF）在舌鳞癌细胞株Tca8113中的表达,为进一步研究iNOS和VEGF在舌鳞癌肿瘤血管生成中的作用提供实验基础。【方法】采用逆转录-聚合酶链反应（RT-PCR）法和SP免疫组化方法,对舌鳞癌细胞株Tca8113中iNOS和VEGF mRNA和蛋白表达情况进行检测。【结果】RT-PCR检测到舌鳞癌Tca8113细胞株中iNOS／VEGF mRNA条带明显,SP免疫组化检测到iNOS／VEGF蛋白在舌癌细胞胞浆中呈强阳性表达。【结论】iNOS／VEGF mRNA和蛋白在舌鳞癌Tca8113细胞株中呈高水平表达。     

大黄素抑制人口腔鳞癌细胞Tca8113增殖及细胞周期进程的实验研究

目的探讨大黄素对人口腔鳞癌细胞体外生长增殖及细胞周期改变的影响。方法采用2.5、5.0、10、20、40、60和80 μmol/L
大黄素分别干预体外培养的口腔鳞癌细胞Tca8113,作用24、48和72 h后,并设含0.1%二甲基亚砜的培养液作为对照组,通过
MTT检测不同浓度大黄素干预对于口腔鳞癌细胞Tca8113的增殖作用;以FCM检测不同浓度大黄素干预对于口腔鳞癌细胞周
期的影响;结合Western blotting 方法分析大黄素干预后对口腔鳞癌细胞周期相关蛋白CDK2、Cyclin E和P21 表达水平的变
化。结果大黄素能够在72 h内明显抑制Tca8113细胞生长以及增殖行为。MTT结果显示,随作用时间从24、48到72 h,大黄素
对于Tca8113细胞的抑制效应呈明显的时间—浓度依赖关系;由细胞周期检测结果显示,大黄素作用使口腔鳞癌Tca8113细胞
周期表现为G0~G1期细胞时相阻滞;蛋白免疫印迹则表明经大黄素干预口腔鳞癌Tca8113 细胞的细胞周期相关蛋白CDK2、
Cyclin E和P21表达水平明显降低,与空白对照组相比较,具有显著的统计学差异（P<0.05）。结论大黄素能够明确抑制人口腔鳞
癌细胞Tca8113的生长增殖以及影响其细胞分裂周期,这可能与抑制细胞周期调控信号通路中分子的活化与转导有关。
     

Toll样受体2对口腔癌Tca8113细胞增殖及凋亡的影响

目的探讨Toll样受体2(TLR2)在人舌鳞癌细胞Tca8113的表达情况及其对肿瘤细胞增殖和凋亡的影响。方法通过流式细胞术检测人舌癌Tca8113细胞中TLR2的表达情况;实验细胞分为实验组(加入TLR2阻断剂)及对照组(加入IgG抗体),实验组及对照组细胞培养24 h及48 h后,应用MTT及Western blot方法检测其增殖及凋亡情况,采用SPSS13.0统计软件对数据进行方差分析及t检验。结果流式细胞检测Tca8113细胞可见TLR2的表达;实验组经TLR2阻断24 h、48 h后其增殖较之对照组明显增加(P<0.05),但24 h与48 h比较,其增殖的变化不明显(P>0.05);同时经Western blot检测抗凋亡蛋白Bcl-2表达明显增加(P<0.05)。结论人舌鳞癌细胞Tca8113细胞表达TLR2受体并对肿瘤细胞呈现明显的抑制作用。     

FasL反义寡核苷酸对舌鳞癌细胞Tca8113增殖和凋亡的影响

目的探讨Fas配体(FasL)反义寡核苷酸(ASO)对舌鳞癌细胞系Tca8113细胞增殖和凋亡的影响。方法设计合成特异性的FasL ASO,并用阳离子脂质体介导转染Tca8113细胞。荧光倒置显微镜观察及流式细胞术(FCM)检测转然前后的转染效率,四甲基偶氮唑蓝(MTT)法检测不同浓度ASO在转染后不同时间对Tca8113细胞增殖的影响;FCM测定转染前后诱导Tca8113细胞的凋亡情况。结果通过荧光倒置显微镜观察及FCM检测发现在脂质体的量一定的情况下,在一定范围内,ASO的浓度越高,转染效率越高。MTT结果显示ASO对Tca8113细胞的增殖有明显抑制作用(P<0.05),FCM对细胞凋亡的检测结果显示反义链组与正义链组、空白对照组及脂质体组相比,总凋亡率有明显差异(P<0.05)。结论 FasL ASO对Tca8113细胞的增殖有一定的抑制作用,能够促进其凋亡,因此FasL ASO有望成为舌鳞癌的潜在的基因治疗方法。     

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.