HPLC法测定生妇宁栓中苦参碱的含量

目的 建立妇宁栓中苦参碱的含量测定方法.方法 采用HPLC法测定苦参碱的含量,流动相为:乙腈-2%磷酸溶液-无水乙醇.检测波长220nm.结果 苦参碱在0.2～1.1μg范围内线性关系良好.加样回收率为97.12%,RSD为0.2986%.结论 本法可用于测定妇宁栓中苦参碱的含量.     

妇洁灵洗剂中苦参生物碱含量变化机制研究

目的 考察苦参及含苦参的妇洁灵洗剂中苦参碱与氧化苦参碱含量变化情况,为临床单用及其在复方中应用苦参提供参考.方法 对妇洁灵洗剂及苦参中的苦参碱及氧化苦参碱采用HPLC进行含量测定,并将处方中共存主要药材与苦参拆方组合,研究其苦参碱与氧化苦参碱的含量变化.结果 所测苦参三氯甲烷提取液氧化苦参碱含量明显高于苦参碱,而苦参水煮后苦参碱含量高于氧化苦参碱;处方中三白草对苦参中苦参碱及氧化苦参碱含量的影响大于蛇床子、火炭母.结论 苦参在单用与在妇洁灵洗剂复方中苦参碱与氧化苦参碱的含量明显不同.妇洁灵洗剂复方煎煮过程中,随着氧化苦参碱的减少,苦参碱的含量逐渐增加.     

高效液相色谱法测定清热通淋丸中苦参碱和氧化苦参碱含量

目的:建立测定清热通淋丸有效成分含量的方法。方法:采用HPLC对清热通淋丸中苦参碱和氧化苦参碱[1]的含量进行测定。结果:苦参碱在0.314 4~3.458μg之间具有良好的线性关系,氧化苦参碱在0.007 4~0.081 9μg之间具有良好的线性关系,平均回收率苦参碱为99.01%,RSD=0.46%(n=6),氧化苦参碱为97.91%,RSD=0.40%(n=6)。结论:该方法灵敏、重现性好,适用于清热通淋丸中苦参碱和氧化苦参碱含量测定。     

艾愈片中苦参碱含量的HPLC测定方法与制备过程中的转化研究

目的:建立艾愈片中的苦参碱含量的HPLC测定方法,考察艾愈片在制备过程中苦参碱含量变化情况。方法:采用SHI-MADU-ODS C18色谱柱(5μm,4.6 mm×250 mm);以乙腈-0.2%磷酸溶液(以三乙胺调至pH 8.0)(30∶70)为流动相;流速为1.0 mL.min-1,检测波长为220 nm;同时考察制备过程中的苦参碱的含量变化。结果:苦参碱在0.164 96~8.248μg之间呈良好线性(r=1.000 0),在艾愈片中的平均回收率为99.67%,RSD为0.7%(n=5)。在水煎煮提取过程中存在氧化苦参碱向苦参碱转化。但在浓缩、干燥及成型过程中不存在苦参碱与氧化苦参碱之间的相互转化。结论:HPLC法可准确、简便、快速测定艾愈片中苦参碱含量,艾愈片制备过程存在氧化苦参碱向苦参碱转化。     

苦参碱和氧化苦参碱体内外模型的肝毒性比较研究

目的比较苦参碱(matrine,MT)和氧化苦参碱(oxymatrine,OMT)的肝毒性,以探索其毒性严重程度和特征,并初步阐明毒性机制。方法用对乙酰氨基酚(acetaminophen,APAP)、苦参碱和氧化苦参碱处理肝细胞,24 h后,检测IC50、肝细胞酶含量、病理形态、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)含量,并检测凋亡率。用对乙酰氨基酚、苦参碱和氧化苦参碱处理斑马鱼,96 h后,检测LC50、肝细胞病理形态、丙二醛和谷胱甘肽含量,并检测凋亡率,同时检测氧化应激相关基因zgc:136383、凋亡相关基因EIF4EBP3和zgc:123120的表达。结果苦参碱和氧化苦参碱对肝细胞有毒性作用,苦参碱IC50为5.3 mmol/L,氧化苦参碱为19mmol/L。苦参碱和氧化苦参碱处理肝细胞的谷丙转氨酶、碱性磷酸酶、谷草转氨酶和乳酸脱氢酶含量升高,肝细胞肿胀,丙二醛含量升高,谷胱甘肽含量降低,凋亡率增加(P0.05)。苦参碱和氧化苦参碱对斑马鱼有毒性作用,苦参碱的LC50为0.41 mmol/L,氧化苦参碱为3.8 mmol/L。苦参碱和氧化苦参碱处理的斑马鱼肝细胞呈现轻度至中度空泡化,丙二醛含量和凋亡率增加,谷胱甘肽含量降低(P0.05)。苦参碱下调氧化应激相关基因zgc:136383(P0.05),下调抗凋亡基因EIF4BP3,上调促基因zgc:123120(P0.05)。结论体内外模型结果一致,苦参碱和氧化苦参碱具有肝毒性,其毒性特征相似,苦参碱的毒性大于氧化苦参碱。肝毒性机制与氧化应激和细胞凋亡有关:苦参碱下调基因zgc:136383减少脂质转运,并激活氧化应激反应;苦参碱上调促凋亡基因zgc:123120,下调抗凋亡基因EIF4EBP3并诱导肝细胞凋亡。     

优化的高效毛细管电泳法测定肝力保胶囊中苦参碱和氧化苦参碱的含量

目的:建立优化的高效毛细管电泳法,并将其应用于测定肝力保胶囊中苦参碱和氧化苦参碱的含量.方法:采用Agilent3DCE高效毛细管电泳仪、未涂渍熔融石英毛细管(68.5 cm×75μm,有效长度60 cm),缓冲液50 mmol/L Na2 HPO4(水:甲醇=4∶1,pH=5.5),运行电压25 kV,温度25℃,检测波长214 nm,以氨茶碱为内标,建立优化的高效毛细管电泳法,并测定3批次肝力保胶囊中苦参碱和氧化苦参碱的含量.结果:苦参碱和氧化苦参碱均能够基线分离,符合含量测定要求;回归方程分别为Y=222.80X 3.635 3(r=0.999 9)和Y=213.02X 34.268 6(r=0.999 9);线性范围分别为33.98～339.8μg/ml和47.35～473.5μg/ml.苦参碱和氧化苦参碱的日内和日间精密度(RSD)均＜3.0%,最低检测限分别为13.60μg/ml和18.90 μg/ml,加样回收率结果分别为102.3%(RSD=0.57%,n=3)和101.1%(RSD=0.41%,n=3).3批样品中苦参碱的含量(%)分别为2.78±0.043、2.50±0.069、2.56±0.040;没有检测到氧化苦参碱的峰.结论:优化的高效毛细管电泳法稳定简便,结果可靠,能够用于测定苦参碱和氧化苦参碱含量;优化的高效毛细管电泳法测定的苦参碱含量可作为肝力保胶囊质量控制指标.     

Determination of Matrine in Baicaofuyanqing Suppository with High Performance Liquid Chromatography

目的:建立用高效液相色谱法(HPLC)测定百草妇炎清栓中苦参碱含量的方法.方法:采用HPLC法,色谱柱为Agilent ZORBAX-NH2(4.6m×250mm,5μm),流动相为乙睛-无水乙醇-1.5％磷酸(80:10:10),流速1.0ml/min,检测波长220nm,柱温35℃.结果:苦参碱在0.0656～0.8528μg与峰面积成线性关系(r=0.9999),该制剂中苦参碱的平均回收率为101.01％,RSD为1.41％(n=6).结论:该高效液相色谱法简便易行、准确、耐用,可用于百草妇炎清栓中苦参碱的含量测定.     

山豆根在复方中化学成分变化的研究

山豆根为豆科植物越南槐 Sophora tonkinensisGagnep.的干燥根及根茎 ,具有清热解毒、消肿止痛、通便之功能 ,主要成分为氧化苦参碱 [1]、苦参碱。在含山豆根药材的中成药的复方制剂中 [2 ]只测定苦参碱含量 ,作者认为山豆根在复方中 ,与共存药物在制备过程中苦参碱、氧化苦参碱可能互有转变。本实验结果表明 ,山豆根饮片用水煎煮 ,氧化苦参碱含量降低 ,苦参碱含量增加 ;山豆根与丹参、木蝴蝶、黄柏、玄参等配伍 ,与水共煎煮 ,氧化苦参碱含量则随时间的延长而减少直至消失 ,苦参碱含量增加。1　实验部分1 .1　仪器、试剂、药品 :CS- 93 0…     

高效液相色谱法测定复方石韦薄膜衣片中苦参碱的含量

目的:建立高效液相色谱法测定复方石韦薄膜衣片中苦参碱含量的方法.方法:采用Kromasil KR100-5C18(4.6mm×250mm,5μm)色谱柱,流动相:乙腈-0.025M磷酸(1:24),流速:1ml/min,检测波长:220nm.结果:苦参碱在16.01μg/ml-320.40μg/ml浓度范围内线性关系良好(r=0.9999),平均加样回收率为100.64%,RSD为0.87%.结论:本文建立的方法简单、灵敏、结果准确,可用于测定复方石韦薄膜衣片中苦参碱的含量.     

苦参汤有效部位总生物碱含量测定方法研究

目的建立苦参汤有效部位中总生物碱含量测定方法.方法采用酸性染料比色法测定总生物碱含量:以苦参碱为对照品,溴麝香草酚蓝为酸性染料,在pH 7.6缓冲溶液条件下,用氯仿萃取.结果苦参碱在1.67～8.33 mg/L范围内与吸光度呈良好的线性关系,回归方程为C=0.076A 0.0068 r2=0.9995.苦参碱平均回收率为103.84%,RSD=0.75%(n=5).结论 5批样品测定结果表明,所建立的方法简便、准确,可用于苦参汤有效部位总生物碱含量测定.     

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.