LC—MS法测定人血浆中地洛他定的浓度及其药代动力学研究

目的　建立 LC-MS测定人血浆中地洛他定浓度的定量方法 ,测定志愿者口服地洛他定胶囊后的血药浓度 ,并对供试制剂与参比制剂的生物等效性进行评价。方法　血浆中加入氟西汀为内标 ,碱化后经乙酸乙酯提取 ,进行 LC-MS测定。色谱柱为 Phenomenex Luna C1 8(5 μm,2 5 0 mm× 4.6mm) ,流动相为甲醇 -乙腈 -0 .0 1mol/L醋酸铵溶液 (65∶ 5∶ 3 0 ) ,流速为 1 .0 ml/min;ESI选择性正离子检测。临床实验方案采用双交差实验设计法。结果　地洛他定血浆线性范围为 0 .1～ 3 0 ng/ml,检测限为 0 .0 5 ng/ml。方法回收率大于 80 %。应用本法测定 2 0名志愿者单剂量交叉口服供试制剂与参比制剂后的血药浓度经时过程 ,测得二者的主要药代动力学参数无显著性差异 ,试验胶囊的相对生利用度为 (96.0 4± 1 1 .1 0 ) %。结论　本方法专属性强 ,灵敏度高 ,准确性好。试验胶囊和参比片剂生物等效     

LC-MS法测定人血浆中格列苯脲及其在健康志愿者中的药代动力学

目的:测定人血浆中格列苯脲的浓度,并研究在中国志愿者中单剂量口服5 mg后的药代动力学过程.方法:用液-液萃取法提取血浆中药物.采用液相色谱-质谱联用的分析手段,建立人血浆中格列苯脲浓度的测定方法.结果:格列苯脲在1.56～400 μg/L浓度范围内线性良好(r=0.9999),其在人血浆中的回收率大于80%,日内、日间RSD分别为3.6%～6.8% 和6.1%～8.4%.最低检测限为0.5 ng/ml.该药在人体内的主要动力学参数为:AUC0-∞=(813.6±254.2)μg·h/L,AUC0-t =(790.6±248.2) μg·h/L,cmax=(130.1±64.8) ng/ml,tmax=(2.6±1.8) h和t1/2=(2.7±1.9) h.结论:本方法专属性强,灵敏度高,可准确测定格列苯脲在人血浆中的浓度.     

阿奇霉素分散片人体生物等效性评价

目的:评价阿奇霉素分散片的人体生物等效性。方法:20名男性健康志愿者随机交叉口服阿奇霉素分散片受试制剂和参比制剂各500 mg,采用高效液相色谱-质谱法(LC-MS)测定血药浓度。以DAS 2.0软件计算其药动学参数,考察其生物等效性。结果:阿奇霉素分散片受试制剂和参比制剂的差异均无统计学意义(P>0.05)。以AUC_0-120h计算,受试阿奇霉素分散片相对生物利用度为(119.6±52.9)%。结论:阿奇霉素分散片受试制剂和参比制剂具有生物等效性。     

HPLC法测定人血浆中丙硫氧嘧啶及药代动力学研究

目的建立人血浆中丙硫氧嘧啶的高效液相色谱测定法,以测定志愿者口服丙硫氧嘧啶片剂后的血药浓度,并对供试制剂和参比制剂的生物等效性进行评价.方法血浆中加入内标磺胺二甲嘧啶后酸化,以乙酸乙酯提取,进行高效液相色谱法检测.色谱柱为KromasilODS150×4.6mm,5μm,流动相为乙腈-水-冰醋酸(15∶85∶1),流速1ml/min,检测波长275nm.20名健康志愿者随机分成两组,分别服用供试片和参比片,临床实验方案采用双交叉实验设计法.结果本测定方法的提取回收率在97.1%～102.1%,用本法测定了20名志愿者随机交叉口服单剂量100mg丙硫氧嘧啶后血浆中药物的浓度经时变化过程,并对其药动学参数进行估算.结论该法简便、准确,测得的2种片剂的主要药代动力学参数无显著性差异.     

盐酸托莫西汀胶囊在健康人体的药动学研究

目的建立快速、灵敏的托莫西汀人体内血药浓度的高效液相色谱串联质谱测定法,并对12名健康志愿者口服盐酸托莫西汀胶囊后在人体内的药动学过程进行研究。方法 12名健康志愿者单剂量口服给药20 mg后,分别于给药前和给药后0.25、0.5、0.75、1、1.5、2、3、4、5、6、8、10、12、24 h采集血样。用高效液相色谱串联质谱法测定血浆中托莫西汀的浓度,并采用DAS药动学程序对试验数据进行处理,求算有关药动学参数。结果单剂量口服给药盐酸托莫西汀胶囊20 mg后,其平均血药质量浓度-时间曲线拟合符合二室模型,T1/2、Tmax、Cmax、AUC0-24、AUC0-∞分别为（4.55±0.79）h、（1.33±0.48）h、（225.18±48.53）μg.L^-1、（971.34±219.52）μg.L^-1.h和（992.60±229.93）μg.L^-1.h。结论建立的托莫西汀人体内血药浓度测定方法灵敏、可靠、简便,盐酸托莫西汀胶囊单剂量给药后在中国健康人体内的药动学行为与国外文献报道基本一致。     

LC-MS法测定人血浆中吡嘧司特钾浓度及药代动力学

目的:建立LC-MS测定人血浆中吡嘧司特钾浓度的方法,测定志愿者口服吡嘧司特钾分散片后的血药浓度,估算其药代动力学参数并对试验制剂与参比制剂的生物等效性进行评价.方法:血浆中加入对乙酰氨基酚为内标,酸化后经二氯甲烷-乙酸乙酯(20∶80)提取,进行LC-MS测定.色谱柱为Shim-pack VP-ODS C18(250 mm×2.0 mm ID),流动相为CH3OH-20 mmol/L NH4Ac,pH 3.0(50∶50),流速0.2 ml/min;ESI选择性正离子检测.临床实验方案采用双交叉实验设计.结果:吡嘧司特钾血浆线性范围为10～1 000 ng/ml,检测限为5 ng/ml.方法回收率大于80%.测定了20名志愿者单剂量交叉口服试验制剂与参比制剂后的血药浓度经时过程,测得试验片与参比片的主要药代动力学参数无显著性差异,试验片的相对生利用度为(99.6±14.6)%.结论:本方法操作简单,专属性强,灵敏度高,准确性好.试验片和参比片生物等效.     

阿奇霉素片的相对生物利用度和生物等效性

目的:考察阿奇霉素片人体相对生物利用度及生物等效性。方法:22名健康男性志愿者,采用交叉给药方案,分别单剂量口服20mg受试阿奇霉素片和参比阿奇霉素片,用液相色谱-串联质谱法测定血浆中阿奇霉素浓度,进行人体相对生物利用度和生物等效性评价。结果:单次口服20mg受试阿奇霉素片和参比阿奇霉素片后,达峰时间Tmax分别为(2.0±1.4)h和(1.8±0.6)h;峰值血药浓度Cmax分别为(584.27±258.93)ng/ml和(523.27±186.53)ng/ml;t1/2分别为(47.64±10.38)h和(51.96±12.49)h;药时曲线下面积采用梯形法计算,AUC0→t分别为(3532.26±1311.69)和(3500.10±1229.70)ng.h/ml,AUC0→∞分别为(3986.30±1443.53)ng.h/ml和(4015.14±1426.56)ng.h/ml。结论:国产阿奇霉素片的相对生物利用度为(104.3±27.8%),主要参数的双单侧t检验,结果显示两种制剂为生物等效制剂。     

高效液相色谱法测定人血浆中氟康唑浓度及其应用

目的:建立测定人血浆中氟康唑浓度的RP-HPLC法,并应用于临床药代动力学研究.方法:采用二氯甲烷萃取,高效液相色谱法检测,以单硝酸异山梨酯为内标,Phenomenx C18色谱柱(4.6 mm×250 mm,4 μm),流动相为乙腈-水-冰醋酸(25:75:0.2),流速为1.0 ml/min;检测波长为261 nm.结果:血浆中内源性物质不干扰氟康唑的检测,氟康唑的线性范围为0.125～8.000 μg/ml,批内、批间精密度均小于15%,低、中、高浓度的(0.25 μg/ml、1 μg/ml和4 μg/ml)平均提取回收率分别为100.0%、98.8%、86.7%.结论:该法操作快速、简单、准确,符合生物样品检测要求,可用于临床药代动力学研究中大批量血样的处理.     

氨基葡萄糖的血药色谱浓度测定

目的:探讨氨基葡萄糖的血药色谱浓度测定方法与效果.方法:选择液相色-质谱联用技术测定血浆中的氨基葡萄糖的浓度.结果:氨基葡萄糖的线性方程Y=6.70 ×10-4X+ 1.11×10-2,线形范围0.10～5.00μg/ml,提取回收率92.1％,在血浆中的平均浓度0.260μg/ml.结论:液相色-质谱联用技术测定血浆中的氨基葡萄糖的浓度方法简单、可行、灵敏度高,具有良好的线性、准确度和精密度.     

高效液相色谱法检测家兔血浆中酮康唑浓度

目的　建立家兔血浆中酮康唑(ketoconazole)浓度的高效液相色谱检测法。方法　采用HypersilODSC18柱,以甲醇 水 三乙胺(79:2 0 :1)为流动相,紫外检测波长2 4 1nm ,安定为内标,乙醚提取,浓缩后进样。结果　酮康唑与内标分离良好;血浆酮康唑浓度在0 .5～10 0 .0 μg/ml范围内线性良好,r =0 .9996 ;最低检测浓度为0 .2μg/ml;0 .5、5 .0、5 0 .0 μg/ml三种浓度的平均回收率分别为91.2 %、96 .9%和98.7% (n =5 ) ;日内和日间RSD <10 %。结论　该方法操作简便、可靠、取血量少,适用于酮康唑的血药浓度测定和临床生物利用度的研究。     

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.     

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.     

Electro-Acupuncture Combined with the Trigger Point Needle-Embedding for Treatment of Primary Trigeminal Neuralgia in 31 Cases

In recent years, the author of this essay has applied electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases, yielding satis- factory results as reported in the following.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Persist in and Carry Forward the Integrative Medical Research——Speech at the 6th National General Congress of Chinese Association of Integrative Medicine by Academician CHEN Zhu,Minister of Ministry of Health

The 6th National General Congress of Chinese Association of Integrative Medicine （CALM） was convened at 19-20, April 2008 in Beijing. Academician CHEN Zhu, the minister of Ministry of Health indicated at the congress that the integration of Chinese and Western medicine is very well in keeping with the situation of our country and the general rule of development in medical science; and as a good integration of Chinese medicine and Western medicine, it is mutually beneficial and advantageous to both of them. Seeing the creativity shown in integrative medical investigation in theoretic and methodological sides, we should and must persist in and develop it.