超高效液相色谱法检测二仙汤中淫羊藿黄酮类成分的含量

目的:建立超高效液相色谱法同时测定二仙汤中淫羊藿主要黄酮类成分的检测方法。方法:采用ZORBAX Eclipse Plus-C18色谱柱(3.0 mm×150 mm,3.5μm);以体积分数0.1%甲酸和浓度5 mmol·L~(-1)乙酸铵水溶液(A相)和甲醇(B相)为流动相进行线性梯度洗脱;梯度洗脱程序:0.0~0.5 min,A相的比例保持95%,0.5~1.5 min,A相的比例由95%降到70%,1.5~3.5 min,A相的比例由70%降到50%,3.5~6.0 min,A相的比例由50%降到30%,6~12 min,A相的比例由30%降到10%,12~15 min,A相的比例由10%降到5%;流速为0.45 mL·min~(-1);进样量:1μl;柱温:40℃;检测波长为270 nm。测定对照品混合液、二仙汤水提取液和体积分数50%乙醇提取液中朝藿定B、朝藿定C、淫羊藿苷以及淫羊藿次苷Ⅱ的含量。结果:超高效液相色谱法检测朝藿定B、朝藿定C、淫羊藿苷以及淫羊藿次苷Ⅱ的日间精密度RSD为0.21%~0.40%,日内精密度RSD为0.54%~0.89%,稳定性RSD为0.26%~0.90%,回收率为104.8%~116.7%。同一提取剂的二仙汤提取液中,不同主要黄酮类成分存在明显的差异,表现为朝藿定C淫羊藿苷朝藿定B淫羊藿次苷Ⅱ含量;二仙汤体积分数50%乙醇提取液中检测的主要黄酮类成分均显著高于二仙汤水提取液,差异均有统计学意义(P0.01)。结论:超高效液相色谱法具有十分良好的精密度、稳定性以及回收率,符合方法学的要求,可用于二仙汤中淫羊藿主要黄酮类成分的含量测定,为二仙汤的质量控制提供依据。     

UPLC法测定不同基源淫羊藿中11种有效成分的含量

[目的]对19批不同基源淫羊藿和4批巫山淫羊藿中的主要有效成分进行含量测定比较.[方法]采用LC30超高效液相色谱仪,Shim-pack GISS C18色谱柱(100 mm×2.1 mm,1.9μm)梯度洗脱,对淫羊藿和巫山淫羊藿中新绿原酸、绿原酸、隐绿原酸、木兰花碱、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、箭藿苷A...     

HPLC法测定不同产地淫羊藿中7种主要黄酮类成分的含量

目的 采用HPLC法测定不同产地淫羊藿药材中7种主要黄酮类成分的含量。 方法 色谱柱为SHISEIDO MG-C18柱 (3.0 mm×100 mm, 3.0 μm);流动相为乙腈(A)和0.1％的甲酸水溶液(B),梯度洗脱,A相含量随时间的变化：25%(0～10 min),25%～40%(10～12 min),40%～45% (12～22 min), 45%～75% (22～25 min), 75%(25～30min);流速0.6 mL/min;检测波长270 nm;柱温25 ℃;进样量5 μL。淫羊藿药材以70%乙醇超声提取。 结果 7种黄酮类成分朝霍定A、朝霍定B、朝霍定C、淫羊藿苷、淫羊藿次苷I、淫羊藿次苷II、脱水淫羊藿素在30 min内基线分离。方法学验证表明,线性关系良好(r＝0.9999),日内日间精密度RSD%均小于2.0%,回收率在98%～102%之间,稳定性和重复性RSD%也均小于2.0%,符合方法学要求。测定了对照药材及辽宁、甘肃、湖北3个产地淫羊藿中7种黄酮类成分的含量。　结论 该方法快速简便,可为淫羊藿药材的质量控制提供依据,也为进一步开展淫羊藿中黄酮类成分的药动学及组织分布研究奠定了良好的基础。     

生长期巫山淫羊藿不同部位5种黄酮类成分的动态积累研究

目的 研究生长期巫山淫羊藿根、茎、叶中5种黄酮类成分朝藿定C、双藿苷A、淫羊藿苷、淫羊藿属苷A和淫羊藿属苷C的量,揭示几种活性成分的动态积累规律。方法 采用RP-HPLC方法,以美国Waters Sunfire TM-18为色谱柱(带预柱),检测波长270 nm,流动相为乙腈-水,梯度洗脱,体积流量为1 mL/min。结果 在20 min内分离测定了5种异戊烯基黄酮,得到了这5种异戊烯基黄酮在生长期巫山淫羊藿不同部位中的分布积累规律。发现巫山淫羊藿不同部位中双藿苷A的量分别为：根1.090%～3.661%,茎0.001%～0.033%,叶0.095%～0.217%;淫羊藿属苷A的量分别为：根0.177%～0.971%,茎0.010%～0.089%,叶0.089%～0.323%;朝藿定C的量分别为：根0.223%～0.748%,茎0.024%～0.147%,叶0.905%～2.228%;淫羊藿苷的量分别为：根0.009%～0.128%,茎0.003%～0.044%,叶0.258%～0.929%;淫羊藿属苷C的量分别为：根0.080%～1.857%,茎0.002%～0.022%^{,叶0.004%～ 0.058%。结论 巫山淫羊藿叶中淫羊藿苷除4、5、7月份外,其余月份的量都低于0.5%,而朝藿定C的量平均在1.586%。巫山淫羊藿叶在其生长的任何月份都是朝藿定C的量远大于淫羊藿苷的量,积累高峰期有3个,朝藿定C是巫山淫羊藿叶中的主要成分。巫山淫羊藿根在其生长的任何月份都是双藿苷A的量远大于朝藿定C的量,质量分数平均在2.763%,双藿苷A为巫山淫羊藿根中的主要成分。}     

UPLC法同时测定淫羊藿中5种黄酮类成分的含量

目的建立同步测定中药材淫羊藿中朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷I 5种黄酮类成分含量的UPLC-DAD法。方法采用Aglientl290型超高效液相色谱仪,色谱柱为Aglient Zorlbax EclipsePlus C_(18)柱(50mm×2.1mm,1.8μm),柱温为40℃。流动相为乙腈和0.1%磷酸水,梯度洗脱,流速为0.4mL/min,进样量为1μL,检测波长为270nm。结果淫羊藿药材中5种黄酮类成分分别在2.02~202μg/mL、2.12~212μg/mL、2.14~214μg/mL、2.52~252μg/mL和1.12~112μg/mL的浓度范围内呈良好的线性关系,最低检测限分别为2.82、3.03、2.72、3.09、1.23ng/mL。重复性、稳定性试验结果的RSD值均小于3%,各成分平均回收率分别为99.76%、98.96%、98.76%、98.67%和98.65%。同时对14批不同药房或产地的药材中5种黄酮类成分的含量进行了检测。结论该方法快速简便、准确可靠,各主要化学成分均能达到基线分离,适用于淫羊藿药材的多成分分析,进而为淫羊藿药材质量控制的全面评价提供科学依据。     

光叶淫羊藿的化学成分研究

目的 研究光叶淫羊藿全草的化学成分,为阐明其有效成分提供依据。方法 利用硅胶柱色谱进行分离,根据化合物的光谱数据鉴定其结构。结果 从光叶淫羊藿根、茎中分离得到了8个化合物,其中1个木脂素类成分,1个甾体类成分,6个黄酮类成分,分别为( 7S,8R,8′R)-3,3′-二甲氧基-4,4′,8′-三羟基-9-乙酰氧基-7,9-环氧木脂素(Ⅰ)、胡萝卜苷(Ⅱ)、淫羊藿次苷C(Ⅲ)、2″-鼠李糖大花淫羊藿苷A(Ⅳ)、淫羊藿苷(Ⅴ)、淫羊藿次苷A(Ⅵ)、朝藿定C(Ⅶ)和柔藿苷(Ⅷ);从光叶淫羊藿叶中分离得到了4个化合物,其中2个黄酮类成分,2个苯乙醇苷类成分,分别鉴定为thalictoside(Ⅸ)、sutchuenoside A(Ⅹ)、icariside D2(Ⅺ)和山柰苷(ⅩⅡ)。结论 所有12个化合物均为首次从该植物中分离得到。     

HPLC法同时测定仙人菇口服液中10种成分的含量

目的：建立快速、准确的高效液相色谱法(HPLC)同时测定仙人菇口服液中腺苷、虫草素、芦丁、芒柄花苷、朝藿定B、朝藿定C、淫羊藿苷、毛蕊异黄酮、灵芝酸A、宝藿苷I等10种有效成分的含量。方法：使用Dikma Diamonsil Plus C18柱(4.6 mm×250 mm,5 μm),以乙腈-0.1%冰醋酸水溶液梯度洗脱；流速为1.0 ml/min,柱温为30°C,检测波长为270 nm,进样量为10 μl。结果：腺苷在0.25~16.0 μg/ml、虫草素和淫羊藿苷在6.25~400.0 μg/ml、芦丁在1.25~80.0 μg/ml、芒柄花苷在0.5~32.0 μg/ml、朝藿定B在0.375~24.0 μg/ml、朝藿定C在3.125~200.0 μg/ml、毛蕊异黄酮和灵芝酸A在0.625~40.0 μg/ml、宝藿苷I在0.125~8.0 μg/ml浓度范围内呈良好的线性关系(r>0.999)；精密度、稳定性、重复性实验的RSD值均小于5%；加样回收率在85%~115%范围内。结论：该法操作简单、准确性高、重复性好,适用于仙人菇口服液中腺苷、虫草素、芦丁、芒柄花苷、朝藿定B、朝藿定C、淫羊藿苷、毛蕊异黄酮、灵芝酸A、宝藿苷I等10种有效成分的含量测定,可以作为仙人菇口服液质量控制方法。     

HPLC测定补肾壮骨颗粒中淫羊藿苷的含量

目的:补肾壮骨颗粒为我院院内制剂,淫羊藿是方中君药.文中采用HPLC建立补肾壮骨颗粒中淫羊藿苷含量的测定方法. 方法:采用HPLC法,色谱柱:Agilent HC-18(150mm×4.6mm,5μm),柱温:30℃,以乙腈-水(30∶ 70)为流动相,流速为1.0ml/min,检测波长270nm,测定了3批补肾壮骨颗粒中淫羊藿苷的含量. 结果:在HPLC色谱中,淫羊藿苷可与其他成分分离,且在0.0962～0.6734μg范围内与峰面积呈良好的线性关系,r2=1(P<0.001),样品平均回收率为94.4%,RSD为1.83%(n=6). 结论:该法分离效果好,分析快速、准确,灵敏度高、重现性好,可用于补肾壮骨颗粒中淫羊藿苷的含量测定.     

补肾强身片中淫羊藿苷的含量测定

目的建立补肾强身片中淫羊藿苷的含量测定方法。方法对国内有五个企业生产的十三个批次的补肾强身片,采用Agilent Eclipse Plus C_(18)色谱柱(4.6×250 mm,5μm),流动相是乙腈-水(30:70),流速1.0 m L/min,检测波长为270 nm,柱温35℃。结果淫羊藿苷回归方程为:Y=20722X+4.495,相关系数r=1(n=6)。淫羊藿苷的线性范围在0.048~0.96μg。结论本方法操作简便、准确、专属性强、稳定,可用于补肾强身片中淫羊藿苷的含量测定。     

Caco-2细胞单层研究淫羊藿黄酮类成分的吸收转运

目的 研究淫羊藿黄酮类成分在Caco-2细胞模型中的吸收转运。方法 采用超高压液相法测定药物浓度,计算表观渗透系数(Papp),研究淫羊藿黄酮5个主要成分淫羊藿苷、朝藿定A、朝藿定B、朝藿定C、宝藿苷Ⅰ在Caco-2细胞模型中的双向转运。结果 淫羊藿苷、朝藿定A、朝藿定B、朝藿定C的吸收渗透系数(P _AB)较小,分别为5.91×10^-7、3.22×10^-7、2.76×10^-7、4.23×10^-7 cm/s,宝藿苷Ⅰ相对较大,为1.46×10^-6 cm/s;5个化合物的分泌渗透系数(P _AB)都比其相应的吸收渗透系数要大,其中宝藿苷Ⅰ的分泌渗透系数是其吸收渗透系数的9.8倍。结论 淫羊藿黄酮类成分的肠道吸收较差,可能存在肠道转运蛋白的外排机制,其中三糖苷(朝藿定A、朝藿定B、朝藿定C)的吸收要小于二糖苷(淫羊藿苷),二糖苷的吸收要小于单糖苷(宝藿苷Ⅰ)。     

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.