HPLC法测定血瘀证患者口服川芎丹参煎剂后阿魏酸血药浓度

目的对以往建立的人血清中阿魏酸浓度测定的HPLC法加以改进,并测定了血瘀证患者口服川芎丹参煎剂后阿魏酸的血药浓度.方法以香豆素为内标,甲醇-水-醋酸(4059.70.3)为流动相,Diamonsil(TM)色谱柱(150 mm×4.6 mm,5 μm),流速1.0 mL/min,以二极管阵列检测器测定322 nm波长下的数据,以沸水浴法加甲醇处理血清样品.结果阿魏酸和内标分离完全,阿魏酸在32.18～965.38 ng/mL范围内线性关系良好(r= 0.998 1),日内和日间RSD＜10%,平均回收率为(99.3±5.4)%,最低检测限为1.6 ng/mL,最低检测浓度为15.7 ng/mL.结论此法简单、快速、灵敏而准确,适用于人血清中阿魏酸血药浓度的测定.     

RP-HPLC测定人血清中阿魏酸浓度的改进方法

目的建立改良的RP-HPLC测定人血清中阿魏浓度的分析方法.方法RP-HPLC法,采用Kromasu-C18柱为分析柱,甲醇-水-冰醋酸(36.4630.6)为流动相,流速1.0 mL/min,检测波长322 nm,内标物为对羟基苯甲醛.用乙腈既去除人血清中的蛋白,又提取血清中的阿魏酸.结果阿魏酸在9.94～1 59.04 ng/mL与峰面积呈良好线性关系(r=0.992 5),日内和日间RSD均<10%,平均回收率为99.77%,最低检测浓度为5 ng/mL.结论该法简便、快速、灵敏、重现性好,低毒,适用于人血清中阿魏酸的定量分析.     

肝郁脾虚证患者口服加味逍遥散后阿魏酸的血药浓度HPLC测定法

目的　建立加味逍遥散汤剂中阿魏酸 (FA)在肝郁脾虚证患者体内血药浓度测定的HPLC法。方法　应用乙腈提取法预处理血样 ,RP -HPLC法测定 10例肝郁脾虚证患者在口服加味逍遥散汤剂后 8个时间点血清样本中的FA浓度。色谱条件 :KromasilC18柱 ( 2 5 0mm× 4.6mm ,5 μm)为固定相 ;甲醇 -水 -冰醋酸 ( 40∶5 9.6∶0 .4,V/V)为流动相 ,流速 :1.0mL/min ;检测波长λ :32 2nm。FA用内标 (对 -羟基苯甲醛 )鉴定。结果　FA的最低检测限为 0 .46 2ng(S/N =3) ,血清最低检测限浓度 4.6 92ng/mL ;线性范围为 7.82～ 195 .5ng/mL(r =0 .9971)。结论　本法简便、快速、灵敏、准确而专一 ,适用于肝郁脾虚证患者口服加味逍遥散汤剂后血清FA的测定。     

高效液相色谱法测定人血清中氨苄西林、丙磺舒的浓度研究

目的:建立测定人血清中氨苄西林、丙磺舒浓度的高效液相色谱法。方法:(1)氨苄西林,以诺氟沙星为内标,血清样品用0.6%的高氯酸沉淀蛋白后进行测定;色谱柱为HigginsTMC18柱(250 mm×4.6 mm,5μm);流动相为有机相(甲醇-乙腈-异丙醇,100∶2∶5,v/v):水相(0.01 mol/L磷酸二氢钾,pH3.5)=23∶77;检测波长210 nm;流速1.0 ml/min;柱温30℃。(2)丙磺舒,以缬沙坦为内标,血清样品用乙腈沉淀蛋白后进行测定;色谱柱为HigginsTMC18柱(250 mm×4.6 mm,5μm);流动相为有机相(甲醇-乙腈,400∶275,v/v):水相(乙酸-水,1∶60,v/v)=77∶23;检测波长280 nm;流速1.0 ml/min;柱温30℃。结果:氨苄西林在0.14~11.2μg/ml浓度范围内线性关系良好(r=0.999 5),最低检测限为0.14μg/ml;丙磺舒在0.20~16.0μg/ml浓度范围内线性关系良好(r=0.999 1),最低检测限为0.20μg/ml。结论:高效液相色谱法操作简便、灵敏、专一性好,适用于人体药动学研究。     

健康人服用黄芪当归合剂后体内阿魏酸的HPLC法测定

目的　建立健康人服用黄芪当归合剂后的血清预处理方法。方法　应用沸水浴一次处理、甲醇二次处理去除血清蛋白质方法 ,用HPLC直接测定健康人服用黄芪当归合剂后体内阿魏酸 (FA)。色谱条件 :甲醇 冰醋酸 水 (4 0∶0 .3∶5 9 .7)为流动相 ,C1 8(Diamonsil)柱 (15 0mm× 4.6mm ,5 μm)为固定相。FA用内标 (香豆精 )法定量。 结果　FA的最低检测限为 2 .5 1ng ;沸水浴 10min的最低检测血清浓度为 2 5 .10ng/mL ,线形范围 40 .16～ 80 3 2 .0 0ng ,r=0 .9975。黄芪当归合剂口服后 5min血清中即能测出阿魏酸 ,在 3 0min达高峰 ,在 90min出现第 2次高峰。结论　该方法保留了水浴法的优点 ,灵敏、快速、简便、专一和重现性好     

灌胃养阴通脑颗粒大鼠血浆中川芎嗪的HPLC法测定

目的建立HPLC测定大鼠灌胃养阴通脑颗粒有效部位组合后血浆中川芎嗪质量浓度的方法。方法以乙腈提取处理血浆,以安眠酮为内标,甲醇-水(55∶45)为流动相,ODSC18色谱柱(150mm×4.6mm,5μm)为固定相,体积流量为1.0mL/min,以紫外检测器在279nm波长下测定血浆中川芎嗪。结果川芎嗪在0.11~22μg/mL线性关系良好,平均回收率为102.98%,RSD<10%,最低检测限为0.21ng,血浆中最低检测质量浓度为0.042μg/mL。结论此法灵敏、可靠,可较好地用于口服养阴通脑颗粒有效部位组合后血浆中的川芎嗪的测定,为测定中药复方体内有效成分提供方法。     

二次萃取法测定人血浆中芬太尼浓度

目的探讨高效液相色谱法-紫外二次萃取法测定人血浆中芬太尼浓度的方法。方法本实验采用内标法,以盐酸瑞芬太尼为内标,以Zorbax-CN(250mm×46mm,5μm)为固定相,以含0.015mol/L磷酸氢二钠和0.05mol/L戊烷磺酸钠的乙腈-水溶液(70:30,v/v)为流动相,流速1.5mL/min,紫外检测波长200nm。结果标准曲线在1～100ng/mL范围内线性关系良好(r=0.999),最低检测浓度为1ng/mL,方法回收率为(91.45±4.55)%,提取回收率为(95.63±4.02)%,日内变异RSD(6.23±2.90)%,日间变异RSD(6.45±4.12)%。结论本方法简便、准确、检测浓度低,能够满足血浆中低浓度芬太尼的测定及临床药代动力学研究的要求。     

健康家兔服用大川芎丸后体内阿魏酸的药代动力学研究

目的建立健康家兔服用大川芎丸后血清中阿魏酸（FA）的高效液相色谱测定法（HPLC）,并研究阿魏酸在大鼠体内的药代动力学。方法用HPLC法,色谱柱为Inertsil ODS-3（5μm,4.6×250mm）,流动相为：甲醇∶水∶冰乙酸（49∶50.745∶0.255,V/V）,流速1.0mL·min^-1,检测波长322nm。FA用内标（香豆精）法定量。结果FA的最低检测限为0.25ng,最低血清检测浓度为5ng·mL^-1（S/N〉3）,线性范围是26.4-4224ng·mL^-1,r=0.9999.平均加样回收率为98.17%±1.26%.结论该方法灵敏、快速、简便、专一性和重现性好,可用于阿魏酸血药浓度分析及药代动力学研究。     

高效液相色谱法测定人肝细胞色素P450 3A4转基因细胞中氨氯地平

目的 :研究氨氯地平经人肝细胞色素 P4 5 0 3A4 (CYP3A4 )的代谢 ,建立反相高效液相色谱的测定人肝转基因细胞 CYP3A4酶 S9孵育液中氨氯地平浓度的方法。方法 :与人肝转基因细胞提取的 S9上清液孵育之后的样品 ,用 3倍量的甲醇沉淀后离心 ,取上清液用 0 .4 5μm微孔滤膜滤过后进样。采用 Hypersil C18柱 ,以乙腈 -磷酸盐缓冲液 (4 5∶ 5 5 ,v/ v,p H4 .5 )为流动相 ,普萘洛尔为内标 ,在 2 5 0 nm波长处测定。结果 :氨氯地平浓度在 0 .2～ 30 .0μg/ ml范围内线性关系良好 ,r=0 .9993,方法平均回收率为 (98.2± 2 .4 ) % (n=5 ) ,日内和日间相对标准偏差(RSD)均小于 10 % ,检测限为 2 0 ng/ ml,定量限为 0 .2 μg/ ml(回收率为 10 4 .0 % ,RSD为 11.4 % ,n=5 )。结论 :建立的反相高效液相色谱方法简便、准确 ,可用于研究氨氯地平在人肝 CYP3A4转基因细胞中的代谢。     

RP-HPLC法测定兔血浆中羟基喜树碱的浓度

目的 :建立测定羟基喜树碱在兔血浆中浓度的反相高效液相色谱 -紫外检测法。方法 :血浆样品酸化后 ,用甲醇沉淀蛋白 ,高速离心后取上清液进行色谱分析。色谱柱 :LichrospherC1 8柱 ( 2 5 0mm× 4.6mm ,5 μm ) ,C1 8预柱 ( 10mm× 4.6mm ,5 μm) ;流动相 :ψ(甲醇 ∶ 0 .0 1mol/L磷酸盐缓冲液 ,pH4.0 ) =60∶40 ;流速 :1.0mL/min ;检测波长 3 84nm。以 5mg/kg剂量给大耳白兔静脉推注羟基喜树碱注射液 ,于不同时间点采血测定药物浓度。结果 :羟基喜树碱保留时间为 5 .0min ,定量线性范围为 5 0～ 5 0 0 0ng/mL ,最低检测限为 2 5ng/mL。血浆中羟基喜树碱的回收率为 99.88%～ 10 3 .13 %,日内精密度≤ 3 .2 7%,日间精密度≤ 7.2 5 %。羟基喜树碱静脉注射后符合二室模型。结论 :本法简便实用 ,定量准确 ,可满足羟基喜树碱药动学研究的需要     

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.