紫杉醇甘草次酸修饰透明质酸纳米粒的制备及性能研究

优化紫杉醇甘草次酸修饰透明质酸（PTX/GA-HA）纳米粒的载药工艺,并系统评价其体内外特性。以载药量、包封率为评价指标,通过单因素考察优化甘草次酸修饰透明质酸纳米粒的载药工艺。制得的纳米粒粒径为（321.2±8.2）nm,荷负电;载药量和包封率分别为（31.2±0.8）%和（90.3±1.6）%。体外释放动力学研究显示,在偏酸性介质中,PTX/GA-HA纳米粒下具有更快的释药速度。同时,MTT实验显示其对多种肿瘤细胞具有杀伤作用,尤其对HepG2细胞的生长抑制作用最强。此外,细胞摄取实验表明,GA-HA纳米粒易被肿瘤细胞摄取。因此,PTX/GA-HA纳米粒具有优良的体内外特性,其成功制备将有助于提高抗肿瘤药物的肿瘤靶向治疗效果。     

pH响应性PTEN/PLGA-(HE)10-MAP纳米粒的构建及体外评价

构建一种pH响应性细胞穿膜肽（cell-penetrating peptides,CPPs）修饰的载抑癌基因第10号染色体同源缺失性磷酸酯酶-张力蛋白（PTEN）质粒DNA的纳米粒PTEN/PLGA-（HE）₁₀-MAP,探讨其基因递送和体外靶向抗肿瘤作用。采用双乳化-溶剂挥发法制备载PTEN质粒DNA的聚乳酸-羟基乙酸共聚物（PLGA）纳米粒PTEN/PLGA;利用酰胺缩合反应将pH响应性组氨酸-谷氨酸（HE）重复寡肽与模型两亲性多肽（MAP）的重组体（HE）₁₀-MAP偶联至PLGA纳米粒表面,得到纳米粒PTEN/PLGA-（HE）₁₀-MAP。以粒径、Zeta电位以及包封率与载药量为指标对其进行表征分析;通过考察其细胞毒性、细胞摄取,以及靶向转染真核表达质粒和抗肿瘤细胞增殖的能力,分析其作为目的基因靶向递送系统的可行性。结果显示,制备的纳米粒粒径为（266.5 ± 2.86） nm,包封率为（80.6 ± 6.11）%,在pH 7.4、7.0和6.5条件下Zeta电位分别为-（6.7 ± 0.26） mV、 +（0.7 ± 0.22） mV和+（37.5 ± 0.85） mV;未载质粒DNA的空载体纳米粒PLGA-（HE）₁₀-MAP在肿瘤和正常细胞中的细胞毒性试验显示细胞存活率均在80%以上,制备的纳米粒可以被细胞摄取表达,且具有pH靶向抑制肿瘤细胞增殖的作用,在肿瘤的基因治疗中具有一定的应用前景。     

O-羧甲基乳糖酰化壳聚糖-聚乳酸阿霉素纳米粒的制备及其物理化学性质的测定

目的制备具有肝靶向性的O-羧甲基乳糖酰化壳聚糖-聚乳酸阿霉素纳米粒,并对纳米粒药物含量、包封率和粒径大小进行检测。方法制备出O-羧甲基乳糖酰化壳聚糖-聚乳酸阿霉素纳米粒,并通过紫外分光光度计测定纳米粒的载药量以及包封率,激光粒度分析仪及电镜测量粒径大小。结果电镜及激光粒度分析仪检测证实纳米粒大小均匀,粒径(197±32)nm,载药量为(44.79±4.27)μg/mg,包封率(67.34±3.32)%。结论该实验制备的纳米粒其粒径小,载药量及包封率高。     

透明质酸修饰的白蛋白纳米粒的制备及抗肿瘤作用的初步评价

目的 制备透明质酸修饰的白蛋白纳米粒,对其修饰程度和载药性能进行考察,并初步评价其抗肿瘤作用.方法 用去溶剂化法制备白蛋白纳米粒,并用透明质酸修饰纳米粒表面,以表面活性氨基的减少作为评价透明质酸修饰的白蛋白纳米粒的修饰程度的指标,并筛选最佳处方;考察pH和载药量及包封率的关系.使用噻唑蓝比色法(MTT)测定纳米粒对人肝癌细胞株HepG2的抑制率.结果 制备所得的透明质酸修饰的白蛋白纳米粒的平均粒径为396 nm,Zeta电位-19.7 mV,表面氨基减少率为34.28%;透明质酸修饰的米托蒽醌白蛋白纳米粒载药量11.13%,包封率94.64%,平均粒径为398 nm,Zeta电位-17.9 mV,且具有明显的缓释作用.透明质酸修饰的米托蒽醌白蛋白纳米粒比米托蒽醌水溶液具有更强的细胞抑制率(P＜0.05).结论 所用制备工艺稳定,可用于制备透明质酸修饰的白蛋白纳米粒.透明质酸修饰的米托蒽醌白蛋白纳米粒具有不低于药物溶液的活性.     

乳铁蛋白修饰水飞蓟素纳米乳的制备及其大鼠体内药代动力学

采用磷脂/吐温80体系制备普通水飞蓟素纳米乳(SM-NE),通过静电作用在其表面吸附乳铁蛋白得到乳铁蛋白修饰的水飞蓟素纳米乳(LF-SM-NE),分别考察两者包封率、形态、粒径、Zeta电位,随后采用LC-MS/MS测定经口给药后大鼠体内的吸收情况。研究结果显示,当乳铁蛋白浓度为0.15 mg/mL,温孵时间为4 h时,乳铁蛋白在SM-NE表面的吸附趋于饱和,所得LF-SM-NE外观圆整呈类球状,平均粒径(257±5.9)nm,Zeta电位-(33.1±1.5)mV,包封率为(96.9±1.9)%,载药量为(3.90±0.12)%;与普通水飞蓟素纳米乳相比,经LF修饰后的SM-NE在大鼠体内的药物浓度显著增高,c_max和AUC均提高约2倍左右。因此,经LF修饰后的SM-NE可进一步提高药物的在体内吸收,是一种新型纳米粒口服吸收增强技术,具有良好的应用潜能。     

烷基化白蛋白的合成、表征以及对难溶性抗肿瘤药物的负载

对牛血清白蛋白进行烷基化修饰,合成一系列两亲性的十二烷基白蛋白(DSA)衍生物,并采用¹H NMR、元素分析以及热重分析法对其进行结构确证;以芘作为荧光探针,测定了它们的临界胶束浓度;采用透析法制备一系列紫杉醇烷基白蛋白(PTX-DSA)纳米胶束,以载药量、包封率、粒径和Zeta电位作为评价指标,研究了取代度对紫杉醇载药能力的影响,然后采用透射电子显微镜(TEM)和广角X线衍射(WAXD)技术分别对载药胶束进行了表征。研究结果表明,取代度在7.72%~31.71%之间的DSA能在水溶液中自组装形成纳米胶束,其紫杉醇的载药量和包封率分别高达（32.14±4.13）%和（87.25±16.18）%,而其粒径和Zeta电位分别为（135.83±2.47） nm和－（31.07±0.51）mV;TEM显示该聚合胶束呈现出球形或类球形结构;WAXD研究结果表明,紫杉醇均匀分散在聚合物胶束中。DSA可作为难溶性抗肿瘤药物的载体,具有载药量高、稳定性好等优点。     

叶酸偶联纳米紫杉醇的制备

目的制备磷脂酰乙醇胺-聚乙二醇2000-叶酸(folic acid monomethoxy-polyethylene glycol 2000-distearoyl phosphatidylethanolamine,ESPE-PEG2000-FA)修饰的紫杉醇纳米脂质体并考察其性质。方法采用超声乳化-溶剂挥发法制备DSPE-PEG2000-FA胶束;采用薄膜分散-挤压法制备紫杉醇脂质体;两者共同孵育形成DSPE-PEG2000-FA修饰的紫杉醇纳米脂质体;并测定修饰前后脂质体的粒径,包封率。结果叶酸偶联纳米紫杉醇药物的粒径(140.5±10.3)nm,多分散指数0.263±0.061,与原料药物纳米紫杉醇的粒径[(121.6±12.7)nm]和多分散指数(0.262±0.031)相比,差异无统计学意义。叶酸-紫杉醇纳米脂质体药物的包封率高达97%,与原料药紫杉醇纳米脂质体的包封率99%相比,差异无统计学意义,保证了药物的纯度和有效性。。结论本研究报道了一种靶向叶酸偶联纳米紫杉醇的新剂型及制备方法。该剂型有良好的药物包封率和胶体稳定性。     

紫杉醇白蛋白片段纳米粒的制备

目的：利用蛋白酶解和纳米粒技术在白蛋白片段的基础上筛选制备紫杉醇纳米粒的新型载体,并考察相关理化指标。方法：利用牛血白蛋白（Bovine serum albumin,BSA）酶解片段作为载体,分散乳化-旋蒸去溶剂法制备紫杉醇-BSA酶解片段-纳米粒,在外观、粒径分布、收率、包封率、载药量及体外释药性方面对样品进行考察。结果：BSA酶解后经粗分离得到4个片段组,选择第4组为候选载体,得到的纳米粒外观形态圆整、均匀,初制备纳米粒混悬剂平均粒径为150.2 nm,平均收率为（45.132±0.903）%,平均包封率为（50.246±0.712）%,平均载药量为（4.804±0.101）%,48 h累计释药82.4%。结论：利用白蛋白酶解片段制备紫杉醇纳米粒是可行的,为扩大紫杉醇纳米粒的载体来源、降低制备成本提供了可能。     

紫杉醇聚乳酸纳米粒的制备及特性考察

目的制备紫杉醇聚乳酸纳米粒并考察其特性。方法采用超声乳化法制备紫杉醇聚乳酸纳米粒,动态光散射粒径仪测定粒径大小,透射电镜表征表面形态,HPLC测定包封率及载药量。结果紫杉醇聚乳酸纳米粒的平均粒径为(230.00±1.08)nm,包封率为(93.57±0.50)%,载药量为(3.01±0.03)μg/mg。结论该方法制备紫杉醇纳米粒工艺简单,方法可行。     

载吲哚菁绿与阿霉素靶向纳米粒用于视网膜母细胞瘤体外成像的实验研究

目的：制备一种靶向叶酸受体的载吲哚菁绿（indocyanine green,ICG）及阿霉素（adriamycin,ADM）的相变型多功能脂质纳米粒（ICG/FA/Pct-PFPNP：ADM）并研究其基本性质,检测其靶向能力、光热效应、超声及光声双模态显像能力,为视网膜母细胞瘤的诊断提供新策略。方法：采用双乳化法制备ICG/FA/Pct-PFPNP：ADM纳米粒,透射电镜观察纳米粒形态,马尔文粒径分析仪检测粒径及电位,紫外分光光度计检测纳米粒的包封率、载药量;流式细胞术量化及激光共聚焦显微镜下观察纳米粒表面叶酸表达情况和体外寻靶能力;CCK-8法检测ICG/FA/Pct-PFPNP纳米粒的生物安全性;近红外激光（808 nm）激发ICG/FA/Pct-PFPNP：ADM纳米粒,检测其超声/光声双模态显像能力。结果：成功制备了ICG/FA/Pct-PFPNP：ADM纳米粒,其外观呈球形,平均粒径为（374.40±23.48） nm,平均电位（-28.0±4.5） mV。测得ICG包封率为（91.85±2.98）%,载药量为（9.19±0.29）%;ADM包封率为（62.04±5.10）%,载药量为（6.20±0.51）%。流式细胞术和激光共聚焦显微镜结果均显示纳米粒表面叶酸表达情况良好,靶向组与Y79细胞连接率为（96.57±3.17）%,与非靶向组、抗体封闭组相比差异具有统计学意义（F=1537.6,P＜0.05）。CCK-8结果显示ICG/FA/Pct-PFPNP纳米粒没有明显的细胞毒性。ICG/FA/Pct-PFPNP：ADM纳米粒经激光辐照后能发生液气相变,增强超声显影并能使局部温度上升至60.8 ℃。光声信号随纳米粒浓度升高而增强,具有良好的线性关系。结论：成功制备了载吲哚菁绿与阿霉素的靶向相变型纳米粒,其靶向能力好,光热效应佳,增强超声及光声显像效果好,可以用于视网膜母细胞瘤的诊断。     

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.