UPLC-MS/MS法同时测定三叶青中13种黄酮类成分的含量

目的:建立超高效液相色谱串联质谱(UPLC-MS/MS)法同时测定三叶青中13种黄酮类成分(原花青素B1、原花青素B2、儿茶素、异槲皮苷、芦丁、山柰酚-3-O-芸香糖苷、紫云英苷、槲皮苷、丁香亭-3-O-葡萄糖苷、阿福豆苷、槲皮素、山柰酚和异鼠李素)的含量.方法:超高效液相色谱采用Waters ACQUITY UPLC BEH C18柱(100 mm×2.1 mm,1.7μm);流动相为乙腈(A)-0.1％甲酸水溶液(B),梯度洗脱;流速0.25 mL/min;柱温40℃.质谱条件采用电喷雾离子源负离子模式进行检测,多反应监测模式定量测定.结果:在考察的浓度范围内,13种黄酮成分均呈良好的线性关系(r≥0.99753);回收率和RSD分别为95.84％～107.38％和0.84％～2.65％.结论:该方法同时测定了三叶青中13种黄酮类成分的含量,方法可行,重现性好,可为三叶青的质量控制提供参考.     

超高效液相色谱法同时测定覆盆子中3种黄酮成分的含量

目的建立同时测定覆盆子中3种黄酮成分含量的超高效液相色谱法(ultra performance liquidchromatography,UPLC)。方法采用BEH C18(2.1mm×100mm,1.7μm)色谱柱;以乙腈-0.1%磷酸水溶液为流动相,梯度洗脱;流速0.12ml/min,检测波长365nm,柱温20℃。结果椴树苷、槲皮素、芦丁分别在1.31～13.08、1.92～19.20、49.20～492.00ng范围内与峰面积呈现良好的线性关系。平均加样回收率分别为99.7%、99.7%、100.2%,相对标准偏差分别为1.03%、0.74%、1.42%。不同产地和采收期覆盆子中3种黄酮成分含量有显著差异。结论该方法简单、可行,可作为覆盆子药材质量控制的参考依据。     

荷叶总黄酮苷质量控制研究

[目的]建立荷叶提取物中黄酮类化合物的高效液相色谱测定方法。[方法]药材经提取水解后用HPLC通过测定其槲皮素的含量来确定荷叶提取物中总黄酮的含量,采用Shim-packCLC-ODS(4.6mm×150mm,5μm)色谱柱,甲醇-水-磷酸(50∶50∶0.1)为流动相,流速0.8mL/min,检测波长360nm。[结果]槲皮素在0.013～0.130μg范围内呈良好线性关系(r=0.9999),荷叶提取物中槲皮素的平均含量为4.94%,RSD=1.67%;平均回收率为99.15%,RSD=0.60%。[结论]此测定方法简便快捷、精密度高、准确性好,可以作为荷叶中总黄酮的含量测定方法,以控制荷叶黄酮苷的质量。     

基于双波长高效液相色谱法同时测定九蒸九晒熟地黄中7种成分的含量

目的 建立同时测定九蒸九晒熟地黄中梓醇、益母草苷、地黄苷A、地黄苷D、毛蕊花糖苷、异类叶升麻苷、5-羟甲基糠醛(5-HMF)等7种成分含量的方法。方法 采用超声法提取九蒸九晒熟地黄炮制过程中每一次蒸晒后的样品,然后应用高效液相色谱(HPLC)法同时测定样品中7种成分的含量;色谱柱为赛默飞C18(250.0 mm×4.6 mm,5μm),流动相为0.1%磷酸(A)-乙腈(B),梯度洗脱,柱温30℃,检测波长203,332 nm,进样量10μL。结果 7种成分在各自线性范围内线性关系良好,平均加样回收率为97.57%～100.58%,RSD为0.60%～1.78%。结论 该方法准确、稳定,重复性好,可用于九蒸九晒熟地黄饮片的质量控制。     

高效液相色谱法同时测定杜仲叶中4种有效成分的含量

目的：测定杜仲叶中4种有效成分的含量。方法采用HPLC测定绿原酸、槲皮素、山奈酚、芦丁的含量,色谱柱为WondaSil C18 Superb色谱柱（4.6 mm ×250 mm,5μm）,流动相：0.4%磷酸溶液（A）-甲醇（B）进行梯度洗脱,流速1 ml/min,进样量10μl,检测波长262 nm。结果绿原酸的质量在0.825~4.123μg（R2=0.9999）范围内呈良好线性关系,平均回收率为101.89%,RSD为0.49%;槲皮素的质量在0.165~0.824μg（R2=0.9998）范围内呈良好线性关系,平均回收率为99.88%,RSD为2.64%;山奈酚的质量在0.021~0.104μg（R2=0.9999）范围内呈良好线性关系,平均回收率为100.0%,RSD为0.89%;芦丁的质量在0.002~0.012μg（R2=0.9991）范围内呈良好线性关系,平均回收率为100.0%,RSD为0%。结论本方法简便、快速、准确、重复性好,适用于杜仲叶中各成分的含量测定。     

HPLC法测定鬼针草不同部位三种黄酮成分含量

目的 :建立同时测定鬼针草中三种黄酮类成分含量的方法 ,并比较鬼针草茎和叶中三种成分的含量。方法 :采用HPLC法对鬼针草中三种黄酮类成分芦丁、异奥卡宁7-0-β-D-葡糖苷和异槲皮苷的含量测定方法学进行考察,并同时分析三种黄酮类成分在鬼针草茎和叶总黄酮中的含量。结果:芦丁在0.010~1.040 mg/m L(r=0.999 9)、异奥卡宁7-0-β-D-葡糖苷在0.012~1.168 mg/m L(r=0.999 5)、异槲皮苷在0.012~1.230 mg/m L线性关系良好;平均回收率(n=6)分别为98.51%、99.21%、98.73%、97.33%、98.92%、98.77%。采用此方法同时测定鬼针草茎和叶中芦丁、异奥卡宁7-0-β-D-葡糖苷、异槲皮苷三种黄酮的含量。结论:建立的HPLC法同时测定鬼针草中三种黄酮类成分含量的方法良好,可用于鬼针草不同部位中黄酮的含量测定。     

荷叶总黄酮苷质量控制研究*

[目的]建立荷叶提取物中黄酮类化合物的高效液相色谱测定方法.[方法]药材经提取水解后用HPLC通过测定其槲皮素的含量来确定荷叶提取物中总黄酮的含量,采用Shim-pack CLC-ODS(4.6 mm×150 mm,5μm)色谱柱,甲醇-水-磷酸(50500.1)为流动相,流速0.8 mL/min,检测波长360nm.[结果]槲皮素在0.013～0.130μg范围内呈良好线性关系(r=0.9999),荷叶提取物中槲皮素的平均含量为4.94％,RSD=1.67％;平均回收率为99.15％,RSD=0.60％.[结论]此测定方法简便快捷、精密度高、准确性好,可以作为荷叶中总黄酮的含量测定方法,以控制荷叶黄酮苷的质量.     

HPLC法同时测定三叶青不同部位中4种成分含量

目的 建立三叶青药材不同部位中4种活性成分的高效液相色谱(HPLC)含量测定方法。方法 采用HPLC法测定绿原酸、异槲皮苷、山奈酚-3-O-芸香糖苷、槲皮素4种成分含量。流动相采用乙腈-0.2%磷酸溶液;检测波长：360 nm;色谱柱：Agilent Extend C₁₈柱;柱温：30℃;流速：1.0 ml/min。结果 绿原酸、异槲皮苷、山奈酚-3-O-芸香糖苷、槲皮素在1.452～145.2μg/ml, 1.245～124.5μg/ml, 0.684～68.4μg/ml, 1.868～186.8μg/ml内与峰面积呈良好的线性关系(R²分别为0.999 7,0.999 5,0.999 6,0.999 3),平均加样回收率分别为98.8%,101.1%,99.4%,99.2%。三叶青不同部位中4个有效成分的含量存在显著差异,其中三叶青根中山奈酚-3-O-芸香糖苷含量最高,三叶青茎中4种成分含量均在根和叶之间,三叶青叶中绿原酸和槲皮素含量最高但未检测到山奈酚-3-O-芸香糖苷。结论 本研究建立的高效液相色谱法能够同时测定三叶青不同部位中4种成...     

HPLC测定银杏提取物中主要黄酮和萜内酯含量的优化方法

目的建立高效液相色谱(HPLC)测定银杏提取物(GBE)中三种黄酮类化合物和四种萜内酯化合物含量的优化方法.方法GBE中黄酮苷水解后稀释,由反相HPLC测定黄酮含量;GBE用甲醇溶解后,经酸性氧化铝柱净化,由反相HPLC测定萜内酯含量.结果槲皮素、山柰素和异鼠李素分别在5~60、5~60、2.5~40ng范围具有良好的线性关系,平均加样回收率(以槲皮素计)为95.4%,相对标准差(RSD)为2.3%;银杏内酯A、银杏内酯B、银杏内酯C和白果内酯在0.5~7.5μg具有良好的线性关系,平均加样回收率分别为97.2%(RSD=2.4%)、99.3%(RSD=1.9%)、97.4%(RSD=1.8%)和 97.9%(RSD=2.5%).结论本法快速、有效,可用于银杏提取物中黄酮和萜内酯的分析.     

HPLC法测定杜仲叶中槲皮素的含量

槲皮素是杜仲叶中一种黄酮类化合物。近年来研究表明，杜仲叶中主要有效成分棚皮素具有抗自由基等作用。因此，测定杜仲叶中槲皮素的含量，可以评价原料的质量。本文建立了HPLC法测定杜仲叶中槲皮素含量的测定方法。1仪器与试药1．1仪器：Varian高效液相色谱仪；2510泵；2550可变波长检测器；4290积分仪；超声波清洗仪YYE－300（北医生产）。1．2试剂与材料：槲皮素对照品购自中国药品生物制品检定所，批号0819003；杜仲叶粉由陕西省略阳杜仲工业公司及贵州遵义安福集团总公司提供。2实验方法与结果21色谱条件：色谱柱：YWG－C18（250m…     

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.