白坚木碱类二聚体在Caco- 2细胞模型中的吸收机制研究

目的 考察白坚木碱二聚体（10- dehydroxyl- 12- demethoxy- conophylline,wtdr- 11）在人类结肠癌细胞系Caco- 2细胞模型中吸收和转运机制。方法 建立Caco- 2单层细胞完整模型,采用超高效液相色谱- 荧光法（ultra- high performance liquid chromatography- fluorescence, UPLC- FLR）测定细胞模型中wtdr- 11的浓度并计算表观渗透系数（apparent permeability coefficient,Papp）;分别考察不同浓度、时间、温度、pH值、P- 糖蛋白（P- glycoprotein, P- gp）抑制剂（维拉帕米和环孢素A）对wtdr- 11在Caco- 2细胞内摄取的影响;考察时间变化对wtdr- 11在Caco- 2细胞中双向转运［分别从细胞刷状缘侧（apical,AP）→细胞基底侧（basolateral,BL）和BL→AP方向］的影响。结果 wtdr- 11的吸收随着时间和药物浓度的增加而增加,但温度、pH值和P- gp抑制剂等因素对Caco- 2细胞吸收该药物的影响无统计学意义（P>0.05）。跨膜转运实验中,Papp＞10-6 cm/s,外排率<1.5。结论 wtdr- 11的主要吸收机制可能是被动扩散,wtdr- 11可能不是P- gp转运蛋白的底物。     

利用Caco-2细胞模型研究新型Pim-1激酶抑制剂KY-C002的表观渗透系数

目的 利用人结肠癌Caco-2细胞模型研究新型Pim-1激酶抑制剂KY-C002的表观渗透系数,为药物制剂设计提供生物药剂学依据.方法 建立Caco-2细胞单层模型,通过细胞形态观察,跨上皮细胞电阻测定和酚红通透性实验验证Caco-2细胞单层完整性.考察不同浓度Pim-1激酶抑制剂KY-C002从绒毛面(AP侧)到基底面(BL侧),BL侧到 AP侧两个方向的转运情况.应用高效液相色谱法对该化合物进行定量分析,计算其表观渗透系数(Papp).结果 建立的Caco-2细胞单层完整,适用于转运实验.Pim-1激酶抑制剂KY-C002浓度为5、10、20 μg/ml时,Pim-1激酶抑制剂 KY-C002 从 AP 侧到 BL 侧的 Papp分别为(4. 72 ± 0.05) × 10-5、(5.29 ± 0.20) × 10-5、(2. 10 ± 0.41) × 10-5 cm/s(P<0.05);从BL侧到AP侧的Papp分别为(7.59±0.78)×10-6、(1.08 ± 0.04) × 10_5、(6.23 ± 0.63) × 10-6 cm/(P<0.05).结论 成功构建Caco-2细胞单层模型并用于考察Pim-1激酶抑制剂KY-C002的透过细胞单层的性能.在考察的浓度范围内,无论从AP侧到BL侧,还是从BL侧到AP侧,Pim-1激酶抑制剂KY-C002的Papp均与其浓度有关,浓度为10 μg/ml时的Papp大于5 μg/ml和20 μg/ml时的 Papp .     

茯苓酸在人源肠Caco-2细胞单层模型的吸收和转运

目的：研究中药茯苓中茯苓酸（pachymic acid，PA）在人肠的吸收和转运。 方法：人源肠Caco－2细胞单层作为药物的肠吸收转运模型，测定PA从绒毛面（AP）到基底面（BL）的渗透性；反相高效液相色谱法测定PA在AP和BL的浓度，紫外检测波长为210nm；计算转运参数和表观渗透系数（Papp），并与在Caco-2细胞单层模型呈良好吸收转运的阳性对照药普萘洛尔和不良吸收转运的阳性对照药阿替洛尔进行比较。 结果：PA由AP到BL方向的Papp值为（9．50±2．20）×10^-7cm／s，由BL到AP方向的Papp值为（11．30±5．90）×10^-7cm／s。在本项研究条件下，普萘洛尔和阿替洛尔的Papp值分别为1．45×10^-5cm／s和4．22×10^-7cm／s。 结论：PA在Caco-2细胞单层馍型的吸收转运呈浓度依赖性，与转运时间呈线性关系；由AP到BL和BL到AP两个方向的转运不良，其Papp值与阿替洛尔的接近；除被动扩散机制外，ATP部分地参与了PA的转运。     

野黄芩苷和野黄芩素在Caco-2细胞模型中的吸收转运特性

目的:研究黄酮类化合物野黄芩苷(scutellarin)和野黄芩素(scutellarein)在Caco-2单层细胞模型中的吸收和转运特性。考察缓冲液、作用时间、P-糖蛋白(P-glycoprotein,P-gp)抑制剂、多药耐药蛋白2(multidrug resistance protein 2,MRP2)抑制剂、乳腺癌耐药蛋白(breast cancer drug resistance protein,BCRP)抑制剂对野黄芩苷和野黄芩素吸收转运的影响。方法:利用Caco-2单层模型研究从绒毛面(apical,AP)侧到基底面(basolateral,BL)侧以及从BL侧到AP侧两个方向的转运过程。应用高效液相色谱法定量分析,计算转运参数和表观渗透系数(apparent permeability coefficient,Papp)。结果:两化合物在磷酸盐缓冲液和Hanks缓冲液中的Papp有差异。在磷酸盐缓冲液中野黄芩苷AP→BL侧无转运,BL→AP侧的Papp为(0.74～1.58)×10-6cm/s;野黄芩素在AP→BL侧与BL→AP侧的Papp分别为(4.33～6.79)×10-6cm/s和(1.3...     

川芎嗪在Caco-2细胞单层模型的转运特征及对P-糖蛋白表达的影响

目的 研究川芎嗪在Caco-2细胞单层模型的转运特征以及对P-糖蛋白（P-gp）表达的影响。方法 MTT法确定川芎嗪对Caco-2细胞单层模型作用的安全浓度范围;以Caco-2细胞单层模型研究川芎嗪的双向转运机制,以表观渗透系数（P_app）为检测指标,考察时间、药物浓度以及P-gp抑制剂维拉帕米对川芎嗪转运的影响;Western blotting法检测川芎嗪对P-gp表达的影响。结果 从顶侧（AP）到底侧（BL）（AP→BL）,川芎嗪的P_app＞10^?6 cm/s,表明其吸收性良好;川芎嗪的转运量与其浓度和时间呈正相关,且川芎嗪AP→BL的转运量明显大于BL→AP的转运量;川芎嗪不仅受到P-gp的外排作用,同时也抑制P-gp表达。结论 川芎嗪在Caco-2细胞模型的转运方式为被动转运,且受到P-gp的外排作用,并对P-gp的表达有抑制作用。     

Caco-2 细胞单层模型中熊果酸摄取转运机制的研究

目的 研究熊果酸在 Caco-2 细胞单层模型中的吸收转运机制。方法 利用人源结肠腺癌细胞系 Caco-2 细胞单层模型研究熊果酸在有或无 P-糖蛋白专属性抑制维拉帕米存在时,评价其双向转运特征,并考察时间、药物浓度、体系温度以及培养介质 pH 值对 Caco-2 细胞摄取熊果酸的影响。采用高效液相色谱-紫外检测法对熊果酸进行定量分析,计算其表现渗透系数 (Papp)。结果 浓度在 10～40 μmol/L 内,Caco-2 细胞对熊果酸摄取量呈线性增加。双向转运研究发现加入 P糖蛋白专属性抑制剂维拉帕米后,其 Papp 发生显著改变,表观渗透率由 3.445 下降至 1.386。结论 熊果酸在 Caco2 细胞模型的吸收转运机制以被动转运为主,P-糖蛋白参与主动转运的过程。     

毛蕊异黄酮体外转运研究及对多药耐药基因MDR1表达的影响

目的研究毛蕊异黄酮在体外模型Caco-2模型的转运特征,及对多药耐药基因MDR1表达的影响。方法采用MTT法确定毛蕊异黄酮对Caco-2模型作用的适用浓度范围;考察浓度、p H、温度及P-糖蛋白(P-gp)抑制剂(维拉帕米)和能量代谢抑制剂(叠氮化钠)对毛蕊异黄酮转运的影响;抑制多药耐药基因MDR1对毛蕊异黄酮转运的影响。结果从上层(AP)到下层(BL),毛蕊异黄酮的Papp10-6cm/s,表明吸收性良好;毛蕊异黄酮的转运与其浓度和时间呈正相关,4℃条件下毛蕊异黄酮的Papp与25、37℃下的Papp差异有统计学意义[AP→BL:(1.85±0.16)×10-6cm/s比(3.05±0.13)×10-6cm/s、(3.89±0.31)×10-6cm/s,P0.01;BL→AP:(1.94±0.29)×10-6cm/s比(3.49±0.18)×10-6cm/s、(4.06±0.26)×10-6cm/s,P0.01];p H=9时的Papp与p H=5、7.4时的Papp值差异有统计学意义[(1.13±0.16)×10-6cm/s比(3.46±0.32)×10-6cm/s、(2.72±0.26)×10-6cm/s,P0.01]。毛蕊异黄酮的转运不受MDR1基因的影响;毛蕊异黄酮的BL侧到AP侧和AP侧到BL侧的Papp比值1.5,不受能量代谢影响。结论毛蕊异黄酮在Caco-2模型的转运方式以被动扩散为主,不受P-gp和能量代谢的影响,且毛蕊异黄酮在低温和碱性环境下不利于其吸收。     

乳糖化-去甲斑蝥素及其纳米粒在Caco-2细胞模型中的跨膜转运机制

目的 研究乳糖化-去甲斑蝥素（Lac-NCTD）及其壳聚糖纳米粒（Lac-NCTD-NPs）的细胞摄取、转运机制。方法 采用Caco-2细胞单层模型研究Lac-NCTD和Lac-NCTD-NPs的摄取和跨膜转运;考察了时间、温度、pH值、药物质量浓度、吸收抑制剂和促进剂对药物跨膜吸收的影响,并比较两种剂型吸收过程的差异。结果 Lac-NCTD以主动转运为主要方式被细胞摄取和转运,少部分通过旁路转运。药物的摄取和时间呈正相关,与温度呈负相关。P-糖蛋白（P-gp）和多药耐药相关蛋白2（MRP2）抑制剂能增加Lac-NCTD的细胞摄取（P＜0.05）。药物从基底侧（basolateral,BL）到肠腔侧（apical,AP）的渗透系数（P_app(BL→AP)）大于P_app(AP→BL)。内吞抑制剂氧化苯砷对药物的转运无影响,旁路转运促进剂去氧胆酸钠能增加药物转运。结论 Lac-NCTD主要以主动转运方式被吸收,少部分通过旁路转运被Caco-2细胞摄取和转运,此过程受P-gp和MRP2外排蛋白作用,且药物纳米粒的摄取和转运较其溶液均有增加。     

塔斯品碱在Caco－2细胞模型中的吸收机制研究

目的研究塔斯品碱在Caco-2细胞模型中的吸收机制。方法用Caco-2细胞单层模型研究塔斯品碱的双向转运,考察时间、药物质量浓度对塔斯品碱吸收的影响。采用高效液相色谱法检测塔斯品碱的质量浓度,计算其表观渗透系数（Papp）。结果塔斯品碱在Caco-2细胞模型中,从单层细胞层顶端到基底端的转运与基底端到顶端的转运大致相同。结论塔斯品碱在Caco-2细胞模型中的吸收主要是被动转运。     

补阳还五汤黄酮类有效成分在Caco-2细胞单层模型的转运特征及对CYP450酶活性的影响

目的研究补阳还五汤3个黄酮类有效成分(羟基红花黄色素A、毛蕊异黄酮、芒柄花素)在Caco-2细胞单层模型中的转运特征及其在大鼠肝微粒体中对CYP450酶亚型活性的影响。方法 MTT法研究药物在Caco-2细胞单层模型的安全浓度范围,以Caco-2细胞单层模型研究黄酮类有效成分的双向转运机制,以表观渗透系数(Papp)为检测指标,考察时间、浓度以及P-gp抑制剂维拉帕米对转运的影响,利用Cocktail探针法研究药物对CYP450亚型的体外抑制作用。结果从顶侧(apical,AP)到底外侧(basolateral,BL)(AP→BL),芒柄花素、毛蕊异黄酮的Papp>10-6 cm/s,表明其吸收性良好,羟基红花黄色素A的Papp为10-6cm/s,表明其吸收性一般;且三者的转运量随浓度升高和时间延长而显著升高,BL→AP的转运量与AP→BL的转运量的比值小于1.5,其中芒柄花素、毛蕊异黄酮的转运都受到P-gp外排蛋白的外排作用;补阳还五汤黄酮类有效成分明显降低了CYP2E1、CYP1A2酶探针底物的代谢量(P<0.05)。结论 3种黄酮类有效成分在Caco-2细胞模型的转运方式均为被动转运,毛蕊异黄酮与芒柄花素明显受到P-gp的外排作用,补阳还五汤黄酮类有效成分可以增加主要通过CYP2E1、CYP1A2酶代谢的药物浓度。     

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.