高效液相色谱法监测环孢素A的全血浓度

目的:建立高效液相色谱法测定环孢素A全血浓度的方法。方法:采用SpherisorbC8(150mm×4.6mm,5μm)分析柱,流动相为甲醇∶水(75∶25,V/V),检测波长214nm,流速1ml/min,柱温为60℃。结果:环孢素A浓度在50ng/ml～2000ng/ml范围内线性良好(r=0.9998),平均方法回收率为99.7%,日内及日间精密度<6%(n=5)。结论:本方法简便、准确、可靠,能较好的满足临床血药浓度监测的需要。     

柱后衍生化HPLC法测定前列地尔乳剂的有关物质

目的：建立测定前列地尔乳剂中有关物质前列腺素A1含量的方法。方法：采用C18色谱柱,磷酸盐缓冲液-乙腈（2∶1,V∶V）为流动相,β-萘酚为内标,氢氧化钾溶液为柱后衍生液,流速：1.0 ml/min,检测波长：278 nm。结果：前列腺素A1在0.5～10.0μg/ml浓度范围内线性关系良好（r=0.999 7）,精密度试验的RSD为1.0%,重复性试验的RSD为1.2%,平均回收率为100.1%,RSD为1.1%（n=9）。结论：该法适用于前列地尔乳剂中前列腺素A1的检测。     

高效液相色谱法测定环孢素A血药浓度

目的建立一种简便可行的HPLC方法测定环孢素A的血浆药物浓度。方法血样经多步处理后用Ba-iAn-C18柱(5μm×250 mm×4.6 mm)分离;流动相为甲醇-已腈-水-异丙醇(25∶18∶57∶1.5,v/v);流速为1.0 mL/min;柱温为70℃;检测波长为λ=208 nm。结果环胞素A(CsA)血药浓度在50~2 000 ng/mL范围内,标准曲线线性关系良好(r=0.998 0);环胞素A最低检测浓度为50 ng/mL,方法绝对回收率为92.1%~95.7%,相对回收率为94.3%~99.8%,样品稳定性良好。结论本法简便、快速、可行,对于移植术后环孢素A血药浓度检测,效果较好。     

反相高效液相色谱法测定环孢素A血药浓度

目的：建立反相高效液相色谱法测定环孢素A（CsA）的方法.方法：采用Spherisorb-C8型色谱柱,以环孢素D（CsD）为内标,甲醇一水（75：25,V/V）为流动相,柱温60℃,检测波长为214 nm.结果：本法CsA和CsD分离度R=1.608大于1.5,分离效果理想.CsA的浓度在27～864 ng/ml内线性关系良好,其中r=0.9992.日内及日间精密度＜5%（n=4）.结论：本法经济、准确,易于普通实验室展开,能够较好的满足临床血药浓度监测的需要.     

山核桃树叶中球松素查耳酮的含量测定

目的：建立用高效液相色谱法（HPLC）法测定山核桃树叶中球松素查耳酮（DHMC）含量的方法。方法：采用Kromasil C18（250 mm×4.6 mm,5μm）色谱柱,以甲醇-水（65∶35,V/V）为流动相,柱温为30℃,流速为1.0 ml/min,检测波长为342 nm,对DHMC的含量进行测定。结果：DHMC进样量在0.02～0.10μg范围内线性关系良好（r=0.999 6,n=6）,精密度RSD为1.1%（n=6）,重复性试验RSD为0.9%（n=6）,加样回收率为101.6%（RSD=1.8%,n=6）,经测定山核桃叶干品中DHMC的含量较高,达0.79%。结论：所建立的HPLC法专属性强,精密度好,结果准确。     

RP-HPLC法测定人血浆中辛伐他汀的浓度

目的建立人血浆中辛伐他汀的HPLC测定方法。方法采用Thermo-C18柱为分析柱;流动相为乙腈:水(60:40,V/V);流速为1.0 ml/min;检测波长为237 nm;柱温:35℃。结果辛伐他汀、洛伐他汀与人血浆中的内源性物质均实现较好分离,在0.5-25.0 ng/ml的范围内线性良好,r=0.9997(n=5),回收率为93.06%,日内及日间RSD均小于3.01%,最低检测限为0.25ng/ml。结论本法取样量小,操作简便快速,适用于临床辛伐他汀的血药浓度监测。     

高效液相色谱法测定血清中氟康唑的药物浓度

目的:建立高效液相色谱法(HPLC)测定血清中氟康唑药物浓度的方法。方法:采用HPLC法,血清样品采用乙酸乙酯进行提取,用氨苯蝶啶做内标,Lichrospher-C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈∶水∶磷酸(22∶78∶0.28,加乙二胺调pH为4),流速1.0 ml/min,柱温20℃,检测波长295 nm。结果:HPLC法在0.1~6.0μg/ml之间线性良好(r=0.999 9),日间精密度和日内精密度相对标准差均10%,低、中、高三个浓度样品平均提取回收率85.93%~100.60%。结论:本方法快速、准确、重现性好,为氟康唑临床血药浓度监测和药代动力学研究提供参考。     

反式白藜芦醇苷在大鼠体内的药物动力学

目的建立反相高效液相色谱法测定大鼠灌胃后血浆中反式白藜芦醇苷的浓度。方法采用色谱柱Hypersil BDS C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-1%冰醋酸溶液(22:78,V/V);流速0.8 ml/min;柱温25℃;检测波长306 nm。结果线性范围0.012625~2.525μg/ml,血浆标准曲线为R=4.264995 17.5689C,相关系数0.9998,日内、日间精密度均小于9%。方法回收率为95.0%~103.0%。根据大鼠灌胃给药50,100,200 mg/kg后测得的血药浓度,用DAS 1.0软件处理后求得的反式白藜芦醇苷的主要药动学参数:AUC(0-2.5)分别为2.637,0.925,0.617 mg/(h.L),Vβ分别为89.490,79.717;31.253 L/kg,CL(s)分别为54.915,95.661,73.631 L/(h.kg)。结论建立了白藜芦醇苷血药浓度的HPLC测定方法,阐明了白藜芦醇苷的药代动力学特征。方法专属性高,操作简便,结果准确。     

HPLC法测定恩格列净有关物质的方法学验证

目的：建立恩格列净原料药纯度及所含杂质的测定方法。方法：采用高效液相色谱（HPLC）法,用Waters symmetry C18色谱柱（250 mm×4.6 mm,5 μm）,以0.01%（V/V）三氟乙酸溶液为流动相A,乙腈为流动相B,线性梯度洗脱：0~10 min,流动相B 32%;10~50 min,流动相B 32%~95%;检测波长225 nm,流速1.0 mL/min,柱温45℃。结果：恩格列净与各杂质的分离度均>1.5,检出限均<9 ng/mL, 线性范围满足定量分析要求;重复性试验相对标准差（RSD）均<5%;供试品溶液和对照溶液在8 h内稳定性良好;各杂质的回收率为88.6%~106.1%,RSD均<10%。结论：本研究建立的方法检测灵敏度高,精密度好,可作为恩格列净原料药有关物质的控制方法。     

高效液相色谱法测定人全血中环孢素的浓度

目的:建立HPLC法测定人全血中环孢素(CsA)的血药浓度,为临床合理应用CsA提供科学依据。方法:全血样品经乙醚萃取后,采用C18柱(250 mm×4.6 mm,5μm)分离;流动相为甲醇-水(87∶13);检测波长为210 nm,柱温60℃,流速为1.0 ml/min,进样量为20μl,内标为环孢素D(CsD)。结果:在50～2 000 ng/ml范围内,CsA峰面积与内标峰面积的比值与浓度线性关系良好(R2=0.9987),平均方法回收率为97.3%～98.9%,平均萃取回收率为73.9%～79.4%;日内、日间RSD均小于5.2%。结论:高效液相色谱法简便、快速、准确可靠,可用于再生障碍性贫血患者的CsA血药浓度监测。     

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.