RP-HPLC法测定朱砂莲胶囊中马兜铃酸A的含量

目的 建立HPLC法测定朱砂莲胶囊中马兜铃酸A的方法。方法 采用RP-HPLC法测定。用MsphereC18色谱柱(250 mm×4.6 mm,5μm),以甲醇-水-冰醋酸(59:40:1)为流动相,体积流量为1.0 mL/min,检测波长为315 nm,柱温为室温,进样量为20 μL。结果 马兜铃酸A的保留时间约为15 min,与其他峰的分离度大于1.5。马兜铃酸A的线性范围为1.33～26.6μg/mL,r=0.999 9,最低检测限为1.33 ng/mL,平均回收率为98.6％(n=5)。结论 该方法简便快速,结果准确可靠,可用于朱砂莲胶囊中马兜铃酸A的测定。     

HPLC法测定导赤散及配伍组血浆中马兜铃酸A含量

目的通过检测导赤散及配伍组血浆中马兜铃酸A的含量,探索导赤散配伍减毒的作用和马兜铃酸A在大鼠体内代谢的时量关系。方法采用HPLC法,色谱柱为Diamon TMC18(250mm×4.6mm,5μm),甲醇-水-冰乙酸(72∶27∶1)为流动相;室温;检测波长为315nm;流速为1.0mL/min。大鼠随机分5组(导赤散组、配伍一组、配伍二组、关木通组和空白对照组),按0.054g/kg的中药煎液灌胃给药。第7天、第15天后采血制备血浆。经HPLC法测定上述5组血浆中马兜铃酸A的含量。结果给药7d后,导赤散组、配伍一组和配伍二组血浆中均未检测出马兜铃酸A,关木通组血浆中马兜铃酸A的含量为63.386μg/mL;给药15天后,关木通组、配伍一组和配伍二组血浆中马兜铃酸A的含量分别为:99.890、28.824、50.156μg/mL。导赤散组血浆中马兜铃酸A的含量低于最低检测限。结论导赤散的配伍减毒作用优于配伍一组、配伍二组;马兜铃酸A在大鼠体内的代谢存在时量关系。     

脑灵片对青木香减毒作用的初步研究

目的本试验通过研究脑灵片对青木香中马兜铃酸A含量的影响，以初步探讨青木香的解毒问题。方法青木香与脑灵片共煎后用RP—HPLC法测定马兜铃酸A的吸收值，观测其含量的变化情况。结果加入脑灵片后，青木香中马兜铃酸A的含量明显降低。结论脑灵片可显著降低青木香中马兜铃酸A的含量。     

高效液相色谱法测定马兜铃酸A在细胞培养基中含量变化*

[目的]建立细胞培养基中马兜铃酸A高效液相色谱(HPLC)法的测定方法,考察在细胞培养条件下马兜铃酸A在培养基中的稳定性。[方法]样品经甲醇沉淀蛋白,10 000 r/min,离心5 min,取上清液进样。采用HPLC法,Waters C18柱(150 nm×4.6 nm,5μm),甲醇-水(含3%的冰醋酸)(70∶30,V/V)为流动相;检测波长260 nm,柱温30℃,流速1 mL/min。[结果]马兜铃酸A在0.5~50 mg/L范围内线性良好,相关系数r=0.999 8;日内、日间精密度相对标准偏差(RSD)分别为0.96%、1.92%;回收率为94.01%~104.34%。[结论]马兜铃酸A在培养基中3 d内含量稳定,该法简便、灵敏、特异性强,适用于细胞培养基中马兜铃酸A含量的测定。     

小青龙膏贴中马兜铃酸A限量检查和盐酸麻黄碱含量测定

目的建立小青龙膏贴中马兜铃酸A限量检查和盐酸麻黄碱含量测定方法。方法薄层色谱法检查马兜铃酸A限量。HPLC法测定盐酸麻黄碱含量,采用Zorbax TC-C18色谱柱(250mm×4.6mm,5μm);以甲醇-0.1%磷酸(10∶90)为流动相;流速:1mL/min;检测波长:207nm。结果细辛药材及制剂中未检出马兜铃酸A。盐酸麻黄碱含量线性范围0.0404-0.404μg(r=0.9999),平均回收率和RSD分别为100.21%和1.82%。3批样品中,盐酸麻黄碱含量分别为4.7273mg/贴、4.7137mg/贴、4.7195mg/贴。结论该方法简便、可靠,可用于小青龙膏贴中马兜铃酸A限量检查和盐酸麻黄碱含量测定。     

HPLC法测定广防己中马兜铃酸I和马兜铃内酰胺I的含量

目的 建立广防己药材中马兜铃酸I和马兜铃内酰胺I的含量测定方法.方法 HPLC法测定广防己药材中马兜铃酸I和马兜铃内酰胺I的含量.采用Aglient C18柱(Extend),以甲醇为流动相A,1%醋酸-0.02%三乙胺水溶液为流动相B,采用梯度洗脱;流速0.8 mL/min;检测波长316 mm.结果 在各自的考察范围内,马兜铃酸和马兜铃内酰胺I的线性关系均良好(r值均大于0.99),平均回收率分别为98.55%和97.20%,RSD分别为1.62%(n:6)和1.93%(n=6).结论 本方法简便、准确、回收率高,可用于广防己药材中马兜铃酸I和马兜铃内酰胺I的含量测定.     

UPLC-Q-Exactive-MS/MS测定伤痛宁胶囊中马兜铃酸Ⅰ的含量

目的 建立UPLC-Q-Exactive-MS/MS测定伤痛宁胶囊中马兜铃酸Ⅰ的含量。方法 采用Hypersil GOLD C₁₈色谱柱(2.1 mm×50 mm,1.9μm),流动相为乙腈和0.1%甲酸水溶液(含5 mmol/L甲酸铵),梯度洗脱,流速0.5 mL/min,柱温30℃。采用电喷雾离子源正离子模式,进行平行反应监测模式,选择质荷比(m/z)359.0→298.0和359.0→296.0离子对进行监测。结果 马兜铃酸Ⅰ在0.06～0.2 ng范围内线性关系良好(R²=0.999 5),精密度和稳定性的RSD值均小于3%,加样回收率均值为104.1%(RSD=4.29%)。不同批次伤痛宁胶囊样品中马兜铃酸Ⅰ的含量均值为0.000 004 90%,从药材到成品的转移率均值为103.7%,RSD值为19.5%。结论 该方法专属性好,灵敏度高,准确可靠、耐用,可以用于伤痛宁胶囊中马兜铃酸Ⅰ的含量测定。     

HPLC法测定寒湿痹颗粒中马兜铃酸A

目的：建立测定寒湿痹颗粒中马兜铃酸A的高效液相色谱的分析方法。方法：色谱柱为Agilent Zorbax SB-C18柱（250mm×4.6mm,5μm）,以甲醇-1%冰醋酸水溶液（55∶45）为流动相,流速为1.0ml/min,检测波长为315nm,建立HPLC检测方法。结果：马兜铃酸A峰面积与进样量在0.0102～0.1025μg范围内具有良好的线性关系（r=1.000）。平均回收率为99.3%（n=6）,RSD=0.41%。本方法按信噪比为2∶1计算最低检测限为2ng。结论：该方法简单,可靠,检测限低,重现性好,可用于寒湿痹胶囊中马兜铃酸A的限量检测。     

UPLC法测定复方南星止痛膏中马兜铃酸A的含量

目的 对复方南星止痛膏中马兜铃酸A进行检测,验证产品的临床安全性.方法 通过固相萃取小柱对制剂中的马兜铃酸A进行富集,应用UPLC对其进行检测.结果 在Waters UPLC的高灵敏度色谱条件下,信噪比为3:1时马兜铃酸A的最低检出限为0.1μg;3批制剂成品及中间体中均未检出马兜铃酸A;3批细辛药材中检出微量马兜铃酸A,平均含量为1.96μ/g,推算至每克制剂中含马兜铃酸A约为0.05μg,远低于口服制剂药品中马兜铃酸A含量不得高于0.5μ/g的标准.结论 以处方中细辛所含马兜铃酸A的研究数据说明,复方南星止痛膏临床应用是安全可靠的.     

HPLC法测定关木通及4种中成药中马兜铃酸A含量

目的 了解几种主要马兜铃属植物药中马兜铃酸A的含量以指导用药。方法 采用HPLC方法，测定了关木通粉剂和水煎剂以及川芎茶调丸、青宁丸、龙胆泻肝丸和纯阳正气丸4种中成药中马兜铃酸A的含量。结果 关木通粗粉中马兜铃酸A含量平均为0．23％；川芎茶调丸、青宁丸和龙胆泻肝丸中的马兜铃酸A含量很低；关木通水煎剂中马兜铃酸A含量约相当于甲醇提取液的1／3。结论 马兜铃酸A含量应被列为马兜铃属药品的质量监控标准之一。     

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.