LC-MS法测定大鼠血浆中EXH-1626的浓度

目的:建立测定大鼠血浆中EXH-1626浓度的液相色谱-质谱(LC-MS)方法?方法:大鼠血浆样品中加入内标卡马西平,经蛋白沉淀剂(甲醇∶5%硫酸锌溶液 = 70∶30,V/V)沉淀血浆蛋白后取上清液5 μl进行LC-MS测定?色谱条件:色谱柱为汉邦ODS C18柱(5 μm,150 mm × 4.6 mm),流动相为乙腈∶10 mmol/L醋酸铵水溶液(用乙酸调pH至3)70∶30,流速为0.6 ml/min,柱温30℃;质谱条件:采用电喷雾离子化(ESI)方式,以选择性离子监测(SIM)方式检测,EXH-1626和内标选择检测离子的质荷比(m/z)分别为453.3([M+H]+)和274.3([M+H]+)?结果:大鼠血浆中EXH-1626在5~10 000 ng/ml内线性关系良好(r = 0.999 7),最低定量限(LOQ)为5 ng/ml,由低到高(10?100?1 000和10 000 ng/ml)4个浓度的日内和日间精密度RSD均< 10%,绝对回收率均> 70%,相对回收率90%~110%?结论:该方法快速?准确,灵敏度高,操作简便,适用于EXH-1626血药浓度测定及其非临床药代动力学研究?     

反相高效液相色谱法测定人血浆中奥美拉唑的浓度

目的建立测定奥美拉唑血药浓度的反相高效液相色谱法,考察样品提取及色谱分离条件.方法血浆中加入内标尼美舒利后,用以乙醚提取,进行高效液相色谱法检测.以Dikma Technologies C18柱,甲醇:水:三乙胺:冰醋酸(60:40:0.5:0.5)为流动相进行色谱分离.流速1.0ml/min,检测波长302nm.结果奥美拉唑浓度在50～1 800ng/ml范围内线性良好,r=0.9995.测定浓度为15、100、1 500ng/ml的血浆样品,其日间及日内的RSD分别为0.86%、6.7%、4.93%和3.04%、4.25%、5.22%(n=5).3种浓度的提取回收率分别为77.74%、79.44%和86.30%.结论方法简便、准确、灵敏,可用于奥美拉唑血药浓度的测定.     

LC—MS法测定健康受试者血浆中辛伐他丁的浓度

目的研究健康受试者血浆中辛伐他丁浓度的LC-MS测定方法.方法血浆中辛伐他丁经乙酸乙酯提取后采用选择性离子检测方法测定其血药浓度,色谱柱为Kromasil5μm,4.6mm×150mm;流动相甲醇-水(88∶12).结果辛伐他丁的线性范围为0.1ng/ml～20.0ng/ml,日内和日间精密度均小于15.0%,方法的回收率为89%～102%.结论该测定方法学考察均符合血浆样品的要求,可以运用于临床药物动力学研究.     

高效液相色谱-大气压化学电离源-质谱法测定人血浆中帕洛诺司琼浓度

目的:建立液质联用法(HPLC-MS/MS)测定人血浆中帕洛诺司琼浓度的方法?方法:色谱条件为Hypersil GOLD(150.0 mm × 4.6 mm,5 μm)柱,流动相为0.2%醋酸铵水溶液(含0.1%甲酸)-甲醇(50∶50),质谱条件为大气压化学电离源(APCI源),血浆样品中加入内标,碱化后用乙酸乙酯提取,浓缩后采用高效液相色谱-质谱联用进行测定?结果:血浆样品中帕洛诺司琼浓度在0.04~20.00 ng/ml的线性范围内关系良好(r = 0.999 9),定量下限为0.04 ng/ml;低?中?高浓度的回收率?批内及批间精密度均符合生物样本测定方法学要求?结论:本方法灵敏度高?专属性强?重现性好,可用于帕洛诺司琼药代动力学研究及血药浓度监测?     

反相高效液相色谱法测定大鼠血浆中替米沙坦的浓度

目的 :建立测定大鼠血浆中替米沙坦浓度的反相高效液相色谱法。方法 :血浆样品用乙酸乙酯提取 ,选用LichrospherC18(5 μm ,15 0mm× 4 6mm ,ID)为分析柱 ,甲醇 - 0 1mol/L乙酸铵水溶液 (70∶30 )为流动相 ,流速 1 0ml/min ,对羟基苯甲酸丁酯为内标 ,检测波长 2 97nm。结果 :替米沙坦线性范围为 0 0 16～ 1 2 80 μg/ml,最低检测限达 5ng/ml (S/N >3)。样品平均回收率在 90 %以上 ,日内、日间精密度的RSD均小于 10 %。结论 :此方法可用于替米沙坦血药浓度分析及药代动力学研究。     

高效液相色谱法测定人血浆中盐酸吡格列酮浓度

目的 :建立盐酸吡格列酮血药浓度的测定方法。方法 :采用高效液相色谱法测定血药浓度 ,血样用乙醚萃取 ;色谱柱为HiQsilC1 8(4 .6mm× 1 5 0mm ,5 μm) ;流动相为 0 .0 5 %磷酸二氢钾缓冲液 (pH =6.3 0 )∶甲醇∶乙腈 (3∶2∶3 .5 ) ;流速 1 .0ml/min;测定波长 2 3 0nm。结果 :血药浓度线性范围为 2 5ng/ml～ 3 2 0 0ng/ml(γ =0 .9994,n =1 2 ) ,最低检测浓度为 2 5ng/ml;高、中、低 3种浓度的萃取回收率分别为 :70 .63 %± 2 .61 % ,76.0 5 %± 4.3 1 % ,82 .1 0 %± 6.92 % ;方法回收率分别为 :1 0 1 .2 4%± 6.74% ,1 0 6.1 6%± 7.89% ,90 .77%± 2 .87% ;日内相对标准偏差 (RSD) <7.41 % (n =5 ) ,日间RSD <5 .2 4% (n =5 )。结论 :该法为盐酸吡格列酮的药物动力学研究提供了一种检测方法。     

兔血浆中顺式阿曲库铵的HPLC-荧光法测定

目的:建立以反相高效液相色谱-荧光法测定兔血浆中顺式阿曲库铵(Cisatracurium)浓度的方法.方法:色谱柱为Agilent Hypersil ODS-C18柱(5μm,4.0mm×125mm),流动相为50mmol/L磷酸二氢钾缓冲液(pH3.0)-乙腈(50:50),荧光检测的激发波长200nm、发射波长320nm.结果:顺式阿曲库铵检测浓度在25～2 000ng/ml线性关系良好,r=0.999 2.最低检测限为25ng/ml,日内精密度RSD≤2/56%,日间精密度RSD≤3.68%;提取回收率为85.2%～86.1%(n=5),加样回收率为93.4%～98.8%.结论:本方法简便,灵敏度高,重现性好,可满足药代动力学研究的需要.     

反相高效液相色谱法临床监测心脏瓣膜置换术后患者血浆华法林的含量

目的 建立心脏瓣膜置换术后患者血中华法林浓度的快速、准确的高效液相色谱法(HPLC),便于华法林临床药物浓度监测,实现个体化给药.方法 收集西南医院、重庆医科大学附属第一医院人工机械瓣膜置换术后口服华法林抗凝治疗达稳态的患者100例的血浆样品,经正己烷-二氯甲烷(9:1)萃取富集处理后,在310 nm紫外检测波长;流动相:甲醇:水=75:25(含0.2 ml冰乙酸);流速:0.9 ml/min;柱温:25℃条件下,经过Hypersil BDS C18(200 mm ×4.6 mm,5μm)色谱柱分离测定.结果 华法林的保留时间为6.41 min,样品浓度线性方程为C=3.12 ×10-3·R ±0.038,r=0.9998,线性范围为20～2000 ng/ml,平均回收率为97.5%,日内与日间相对精密度均小于5%.对重庆地区100例心脏瓣膜置换术后患者血浆华法林测定,其血药浓度是( 845.48±388.16) ng／ml.结论HPLC法能快速、准确地测量心脏瓣膜置换术后患者血中华法林浓度,并能进行临床监测.     

反相高效液相色谱荧光法测定人血浆中酒石酸罗格列酮含量

目的:建立快速灵敏的高效液相-荧光检测法测定罗格列酮的血浆浓度。方法:血浆样品加入内标(苯丙哌林)后用乙醚提取,乙醚层经空气流吹干后用流动相定容,20μl进样。色谱柱为KromasilC18(250×4.6mm,5μm),流动相采用0.025mol/L,pH=4.85的磷酸二氢钠缓冲液-乙腈(50:50),流速为0.5ml/min,荧光激发波长为250nm,发射波长370nm,内标法定量。结果:本法的最低定量浓度为4.87ng/ml,在14.6￣608.3ng/ml浓度范围内线性良好,r2=0.999。日内精密度为2.65%￣8.25%,日间精密度为5.0%￣8.6%,萃取回收率为68.04%以上,准确度为85.11%￣103.50%,整个分析过程在15min内完成。结论:该法准确、精密度高、重现性好,适用于人血浆中罗格列酮的动力学研究和生物等效性研究。     

反相高效液相色谱法同时测定血浆中奥美拉唑和兰索拉唑

目的:建立用反相高效液相色谱法(RP-HPLC)同时测定血浆中奥美拉唑和兰索拉唑的方法.方法:血浆中加入奥美拉唑、兰索拉唑和内标泮托拉唑,以乙酸乙酯提取,进行RP-HPLC检测.色谱柱为Shimpark ODS 6 mm×150 mm,10 μm,流动相为甲醇 - 水(含1%三乙胺,用高氯酸调pH至7.0;60∶ 40,V/V),流速为1 ml/min;岛津SPD-6AV可见紫外检测器,检测波长为302 nm,AUFS为0.01.结果:奥美拉唑和兰索拉唑与内标泮托拉唑达基线分离,血浆中奥美拉唑在10～1 000 μg/L浓度范围内线性关系良好,r=0.999 9,日内、日间精密度RSD均小于10%,最低检测浓度为5 μg/L.血浆中兰索拉唑在30～1 000 μg/L浓度范围内线性关系良好,r=0.999 9,日内、日间精密度RSD均小于10%,最低检测浓度为20 μg/L.结论:该法简单、准确、灵敏,适用于对奥美拉唑及兰索拉唑血药浓度的测定.     

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.