As4S4/Mn0.5Zn0.5Fe2O4复合纳米粒的制备及其杀伤宫颈癌Hela细胞的实验研究

目的:通过制备As4S4/Mn0.5Zn0.5Fe2O4 复合纳米粒研究体外热化疗对宫颈癌Hela细胞株的作用?方法:采用改良的化学沉淀-浸渍法制备As4S4/Mn0.5Zn0.5Fe2O4复合纳米粒,通过扫描电镜?能谱仪?原子荧光光谱仪来表征;检测As4S4/Mn0.5Zn0.5Fe2O4 复合纳米粒的释药水平及在交变磁场中的升温能力;通过MTT比色法和流式细胞仪检测化疗组?热疗组及热化疗组对宫颈癌的治疗效果?结果:制备的复合纳米粒粒径为20~40 nm;复合纳米粒体外升温能达到肿瘤的有效治疗温度(41~46℃);释药缓慢,48 h释药为13.28%?在MTT实验中,As4S4/Mn0.5Zn0.5Fe2O4 复合纳米粒组细胞抑制率明显高于单纯纳米雄黄溶液组和Mn0.5Zn0.5Fe2O4 联合交变磁场加热组 (P < 0.05);在凋亡率检测中,As4S4/Mn0.5Zn0.5Fe2O4复合纳米粒联合磁流体热疗组细胞凋亡率明显高于单纯纳米雄黄溶液组和Mn0.5Zn0.5Fe2O4 联合交变磁场加热组(P < 0.05)?结论:采用改良的化学沉淀-浸渍法可以成功制备As4S4/Mn0.5Zn0.5Fe2O4 复合纳米粒,体外实验证明该复合纳米粒联合交变磁场热疗对宫颈癌细胞具有很强的生长抑制和诱导凋亡作用?     

PEG-PEI/Fe_3O_4纳米磁流体-TK对裸鼠移植性肝癌细胞靶向杀伤作用研究

目的研究PEG-PEI／Fe3O4纳米磁流体-TK对人肝癌细胞系SMMC-7721裸鼠移植瘤模型的靶向性治疗作用。方法建立人原发性肝癌稞鼠皮下移植瘤模型，将荷瘤裸鼠随机分成肿瘤对照组、纯自杀基因组、PEG-PEI／Fe3O4纳米磁流体-TK组3组。对照组不作任何处理，实验组于瘤体内分别直接注射纯自杀基因、PEG-PEI／Fe3O4纳米磁流体-TK，同时于裸鼠腹腔内注射GCV，观察不同时段皮下肿瘤的生长情况并做病理学检查，免疫组化法检测肿瘤微血管密度（MVD）以及血管内皮细胞生长因子（VEGF）的表达量，原位末端标记（TUNEL）法检测细胞原位凋亡。同时，用RT-PCR检测肿瘤鼠心，肝，肾的自杀基因表达情况。结果裸鼠皮下成瘤率100％；研究PEG-PEI／Fe3O4纳米磁流体-TK组的肿瘤体积、血清AFP含量、肿瘤MVD和VEGF表达强度均明显低于对照组、纯自杀基因组（P〈0．05），细胞凋亡指数都明显高于后两组（P〈0．05），可见较多的凋亡细胞。RT—PCR测示肿瘤鼠心，肝，肾未见自杀基因表达，而肿瘤细胞则出现自杀基因表达。结论PEG-PEI／Fe3O4纳米磁流体-TK可显著抑制肿瘤的生长，并对肿瘤治疗具有靶向性，是一种较为理想的肝脏肿瘤靶向给药系统，有望成为治疗原发性肝癌的新型生物制剂之一。     

DMSA@γ-Fe2O3纳米磁流体联合卡氮芥在交变磁场下对人胶质瘤细胞U251的影响

目的 研究在一定强度交变磁场下DMSA@γ-Fe2O3纳米磁流体热疗联合化疗药物卡氮芥对人胶质瘤细胞U251的影响.方法 对Fe2O3纳米材料进行体外升温实验,以期找到热疗(43℃)所需的最佳浓度.MTT法评价Fe2O3纳米材料对人胶质瘤细胞U251的毒性作用,并确定卡氮芥对该细胞的工作浓度.将人胶质瘤细胞U251分为对照组、磁纳米热疗组、化疗组、磁纳米热疗联合化疗组(简称热化疗组),对照组更换培养液后继续培养,磁纳米热疗组加入适当浓度的Fe2O3浸提液置于磁场下43℃热疗2h,化疗组加入工作浓度的卡氮芥后继续培养24h,热化疗组加入上述浓度的卡氮芥后置于于磁场下43℃热疗2h,采用流式细胞仪观察各组细胞的凋亡情况.结果 Fe2O3纳米材料升温效应良好,当Fe2O3浓度≥8g·L-1时,温度可达43℃以上.Fe2O3纳米材料对U251细胞毒性为0～1级,均属对细胞无毒性范畴.以24h 时IC10～IC25的药物浓度作为实验的工作浓度,确定卡氮芥的工作浓度为0.02g·L-1;43℃单独热疗和单独化疗均对胶质瘤细胞有抑制和杀伤作用(P<0.05),而在此温度下磁纳米热疗与药物联合抑制作用均明显强于单独热疗和单独化疗(P<0.05);流式细胞仪结果显示,对照组、磁纳米热疗组、化疗组、热化疗组细胞凋亡率分别为2.58%、13.75%、17.60%和35.27%.结论 交变磁场作用下,磁纳米热疗(43℃)可以明显增强化疗药物卡氮芥对人胶质瘤细胞U251的抑制作用.     

Fe2O3 纳米磁流体热疗治疗小鼠结肠癌实验研究

目的研究Fe2O3纳米磁流体热疗对小鼠结肠癌的治疗效果。方法磁流体直接局部注射到小鼠皮下肿瘤组织,交变磁场下加热至43℃,计算瘤体积抑制率,肿瘤组织及各个脏器做病理检测。结果 Fe2O3纳米磁流体在交变磁场(alternatingmagnetic field,AMF)中升温迅速。热疗组相比磁流体对照组、磁场对照组、空白对照组,热疗6天后瘤体体积抑制率分别为50.87%、50.50%、76.57%,P<0.05,差异有统计学意义。病理检测证实肿瘤组织部分凋亡,而正常脏器无变化。结论 Fe2O3纳米磁流体热疗可抑制小鼠结肠癌的增长,对结肠癌的治疗有一定作用,但并不能完全抑制肿瘤的生长。实验证明热疗安全可靠。     

纳米As2O3磁性脂质体的制备及表征

目的:制备将药物治疗与热疗相结合的靶向抗癌药物新剂型-As2O3磁性脂质体。方法:用改良的湿化学法制备MnxZn(1鄄x)Fe2O4(锰锌铁氧体)纳米磁性粒子,运用透射电镜和PE热分析系统进行表征,在高频交变磁场下进行体外加热试验,MTT实验检测其细胞毒性;采用薄膜-超声法加高速搅拌制备纳米As2O3磁性脂质体,用透射电镜、图像分析系统和能谱仪对其进行表征,原子荧光光度计检验脂质体中As2O3的包封率。结果:MnxZn(1鄄x)Fe2O4纳米磁性粒子近似球形,粒径20～40nm,无细胞毒性,居里温度随锰锌比例的不同分布在97℃~140℃之间,其磁流体在高频交变磁场下可升温至35℃~47℃并保持恒定。以此磁性材料为载体制成的As2O3磁性脂质体的平均粒径为(182±125)nm,其中含有As2O3和MnxZn(1鄄x)Fe2O4的成份,药物包封率达到71.16%。结论:MnxZn(1鄄x)Fe2O4纳米粒子是制备医用磁性脂质体的良好载体,采用薄膜-超声法加高速搅拌可制备纳米级As2O3磁性脂质体。     

槐耳清膏诱导胃癌SGC-7901细胞凋亡的实验研究

目的研究槐耳清膏对胃癌SGC-7901细胞增殖抑制和凋亡诱导的作用,并探讨其作用机制。方法用终浓度为0、0．5、1、2、4、6mg／mL的槐耳清膏和10μg／mL的5-氟尿嘧啶（5-Fu）作用于SGC-7901细胞,分别于24h和48h后采用MTT法检测细胞增殖抑制率;48h后收集各组胃癌SGC-7901细胞,琼脂糖凝胶电泳检测DNA,流式细胞术检测细胞凋亡率,RT-PCR检测survivin mRNA表达。结果MTT法检测显示,槐耳清膏对胃癌SGC-7901细胞有增殖抑制作用,并存在浓度和时间依赖关系（P〈0．05）;槐耳清膏6mg／mL组的抑制率48h后达到（77．9&#177;2．3）％,5-Fu组为（53．4&#177;1．6）％,差异有统计学意义（P〈0．05）。流式细胞术检测显示,槐耳清膏4mg／mL组的细胞凋亡率最高,早期凋亡细胞达33．2％;6mg／mL组早期凋亡细胞6．3％,而晚期凋亡细胞为19．9％。RT—PCR结果显示,槐耳清膏组胃癌SGC-7901细胞survivin mRNA表达下调。结论槐耳清膏在体外对胃癌SGC-7901细胞有明显的增殖抑制和凋亡诱导作用,初步推断槐耳清膏诱发胃癌细胞凋亡与其对survivin基因表达的抑制有关。     

PEG-PEI／Fe3O4纳米磁流体-TK对肝癌细胞HepG2凋亡的影响

目的:探讨PEG-PEI/Fe3O4纳米磁流体-TK对肝癌细胞HepG2凋亡的影响.方法:构建重组质粒PEGFP-AFP-TK并通过纳米磁流体分别转染入AFP表达阳性的肝癌细胞HepG2及AFP表达阴性的肝癌细胞SMMC7721.转染12 h后,于荧光显微镜下动态观察;48 h后,RT-PCR和Western blot分析HepG2细胞巾HSV-TK基因的表达情况,用MTT法检测HepG2细胞的存活,用流式细胞分析HepG2细胞的凋亡.结果:纳米磁流体能将重组质粒PEGFP-AFP-TK转入HepG2细胞,并且其转染效率高于脂质体;RT-PCR及Western blot证明转染的HepG2细胞有HSV-TK基因的表达;MTT及流式细胞分析说明HSV-TK基因发挥杀伤细胞作用.结论:纳米磁流体能将重组质粒PEGFP-AFP-TK转染HepG2细胞并获得表达,PEG-PEI/Fe3O4纳米磁流体-TK可显著抑制HepG2细胞增殖,可望成为肝癌基因治疗的新型生物制剂之一.     

As2O3磁性Fe3O4白蛋白微球的制备与表征

目的 :制备用于肿瘤热化疗和逆转多药耐药的As2O3磁性Fe3O4白蛋白微球并表征。方法:化学共沉淀法制备Fe3O4磁性纳米粒,运用透射电镜、X射线衍射分析进行表征,溶血实验及MTT试验进行毒理学评定。去溶剂化交联法制备As2O3磁性Fe3O4白蛋白微球,运用透射电镜和能谱仪进行表征,在交变磁场作用下进行体外升温试验,体外释药方法研究其释药速率。结果:Fe3O4磁性纳米粒近似球形,粒径约20 nm,无溶血作用,细胞毒性为1级。As2O3磁性Fe3O4白蛋白微球近似球形,大小均匀,粒径约193.4 nm,其不同浓度的磁流体在交变磁场下可升温至39.5~58.0℃并保持恒定。体外释药实验证实As2O3磁性Fe3O4白蛋白微球具有明显的缓释功能。结论:Fe3O4磁性纳米粒作为药物载体具有良好的生物相容性。用去溶剂化交联法可以成功制备出As2O3磁性Fe3O4白蛋白微球,As2O3磁性Fe3O4白蛋白微球释药速率缓慢,为研究肿瘤热化疗和逆转多药耐药提供理论和实验基础。     

雪灵芝水提取物对人胃腺癌MGC-803细胞Bcl-2，Bax基因表达的影响

目的研究雪灵芝水提取物对人胃腺癌MGC-803细胞Bcl-2和Bax基因表达的影响。方法不同浓度的雪灵芝水提取物处理体外培养的MGC-803细胞,采用四甲基偶氮噻唑蓝（MYr）法检测细胞存活水平;应用流式细胞术检测细胞周期和细胞凋亡水平;采用逆转录聚合酶链反应（RT—PCR）和蛋白印迹法（Western Blot）测定人胃腺癌MGC-803细胞的Bcl-2和Bax基因表达情况。结果雪灵芝水提取物作用于人胃腺癌MGC～803细胞后,0．1～0．8mg／mL浓度组细胞存活水平随药物浓度的增大而减小（P〈0．05）;0．8mg／mL浓度组MGC-803细胞G．期比例为（61．23±6．45）％、S期细胞比例为（26．86±3．38）％、细胞凋亡率为（0．07±0,01）％,与对照组相比,P〈0．05;与对照组相比,0．8mg／mL浓度组Bcl-2 mRNA表达降低、Bax mRNA表达增加（P〈0．05）;0．2～0．8mg／mL浓度组Bcl-2蛋白表达降低、Bax蛋白表达增加。结论雪灵芝水提取物在一定浓度范围内可降低MGC-803细胞的存活水平,使细胞发生凋亡,其机制可能为通过下调Bcl-2基因、上调Bax基因表达,促进细胞凋亡发生。     

低氧条件下三氧化二砷诱导人肝癌细胞株HepG2凋亡作用

目的：探讨低氧条件下三氧化二砷（As2O3）诱导人肝癌HepG2细胞凋亡作用。方法：分别在常氧和低氧（CoCl2 200μmol/L）条件下以2、4、8μmol／LAs2O3作用人肝癌Hep岛细胞,24、48、72小时后以MTr法检测细胞增殖抑制率,以AnnexinV／PI双染色法流式细胞仪检测细胞凋亡,分析两种条件下As2O3对细胞增殖抑制率和凋亡率的影响。结果：常氧及低氧条件下细胞增殖抑制率,细胞早期凋亡率均随As2O3浓度增加及作用时间延长而升高（P〈0．05）。相同浓度As2O3相同作用时间,低氧条件下细胞增殖抑制率及凋亡率升高更为明显。结论：在低氧条件下As2O3促进细胞凋亡,抑制肝癌HepG2细胞增殖的作用显著增强。     

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.