超临界流体萃取分析蓬莪术挥发性成分

用超临界流体萃取、毛细管气相色谱及气相色谱—质谱对蓬莪术挥发油进行分析。最优化的超临界流体萃取条件为20．0MPa,55℃,0．5mL乙醇作改性剂,动态萃取40min,CO2用量为12mL,并与索氏萃取、水蒸气蒸馏比较。     

瘤果黑种草子超临界CO2流体萃取物的GC-MS分析

目的：研究瘤果黑种草子超临界萃取物的化学成分。方法：采用气相色谱-质谱（GC—MS）法对瘤果黑种草子超临界CO2流体萃取物的化学成分进行分析,并通过面积归一化法对各色谱峰进行定量分析。结果：从瘤果黑种草子超临界CO2流体萃取物中共鉴定出26种化合物,其中挥发性成分16种,脂肪酸10种。结论：挥发性成分以烯酸、酮为主,脂肪酸以不饱和脂肪酸为主,与其活血通经等药理作用相符。     

HPLC法测定丹参超临界萃取物中丹参酮ⅡA的含量

目的:建立超临界流体萃取(supercriticalfluidextraction,简称SFE)技术和高效液相色谱(HPLC)技术联用测定丹参中主要成分丹参酮ⅡA含量的方法。方法:探讨改性剂、温度、压力等条件对回收率的影响,确定SFECO2萃取丹参中丹参酮ⅡA的最佳条件;并采用HPLC法对萃取成分中丹参酮ⅡA进行定量分析,确定最佳的色谱分析条件。结果:用超临界流体萃取丹参的最佳萃取条件为:萃取压力30MPa,萃取温度45℃;分离釜I压力16MPa,温度40℃;分离釜Ⅱ压力8MPa,温度33℃;CO2流量为3.3L/h:萃取时间6h;改性剂用丙酮,流速为0.3L/h。最佳色谱条件:ODSC18色谱柱(4.6mm×250mm),流动相为甲醇水(V∶V=75∶25),紫外检测波长270nm,流速1.0ml/min。丹参酮ⅡA浓度在23.36μg/ml～116.80μg/ml范围内,峰面积与进样量线性关系良好,平均回收率为99.12%,RSD=1.99%。结论:用SFEHPLC联用提取测定丹参中主要成分丹参酮ⅡA回收率高、方法快速简便、结果准确、灵敏度高、重现性好。     

半边旗中二萜类化合物5F对乳腺癌生长和转移的体内抑制作用

为研究半边旗中二萜类化合物5F在体内对乳腺癌生长和转移的影响及其作用机制,以MDA-MB-231乳腺癌原位移植模型观察5F抗乳腺癌的效应。在治疗的同时,观察5F对裸鼠的不良反应。以RT-PCR、Western blot法检测5F对乳腺肿瘤组织血管内皮生长因子(VEGF)、激酶嵌入结构域受体(KDR)mRNA和蛋白表达的影响。结果显示,5F通过减少原发肿瘤体积和肺转移结节数目,显著抑制MDA-MB-231乳腺癌移植瘤的生长和肺转移。这种抑制转移的作用不依赖于其对原发肿瘤的生长抑制作用。5F对裸鼠的肝肾功能无明显的影响。5F抗MDA-MB-231乳腺癌作用机制与5F下调肿瘤组织VEGF、KDR mRNA和蛋白表达水平有关。     

牡丹皮超临界CO2萃取物质量标准研究

采用超临界CO2 萃取技术对牡丹皮的有效部位进行萃取 ,并对其萃取物进行了鉴别及含量测定等方面的研究。采用薄层色谱法 (TLC) ,以 ψ(石油醚∶苯∶醋酸乙酯 ) =16∶2 ∶ 2 为展开剂 ,ρ =5 %FeCl3 乙醇溶液为显色剂对牡丹皮超临界CO2 萃取物中的丹皮酚进行鉴别 ;并建立以正十八烷为内标 ,采用SE -3 0 ( 3 0m× 0 .2 5mm× 0 .2 5 μm)石英毛细管色谱柱 ;进样口温度 2 10℃ ,检测器温度 3 0 0℃的气相色谱测定丹皮酚含量的方法。丹皮酚在 ρ =99～ 693 μg/mL浓度范围内 ,峰面积与内标峰面积之比值与浓度呈良好的线性关系 (r =0 .9999) ;平均回收率为 97.12 % ,RSD =1.2 3 % (n =6)。方法准确、简便、灵敏、快速     

超临界流体萃取-胶束电动毛细管色谱法测定银杏叶粗提物中槲皮素和山奈素的含量

目的 :以超临界流体萃取 ( SFE)银杏叶粗提物中槲皮素和山奈素 ,以胶束电动毛细管色谱法 ( MECC)测定这两种成分的含量。 方法 :采用正交设计法考察 SFE提取工艺 ,SFE条件 :压力 42 m Pa;温度 6 0℃ ;静态萃取时间 4m in;动态萃取体积 4m l;改性剂 0 .2 ml乙醇。 MECC条件 :空心熔融石英毛细管柱 ( 5 5 cm× 75μm ) ;缓冲液为磷酸二氢钠 -硼砂 =2 5 - 6 .2 5mm ol/L ( p H 8.5 ) ;SDS 35 mmol/L ;检测波长 2 5 4nm。结果 :SFE时改性剂对萃取效率的影响最大 ;MECC以肉桂酸为内标 ,在 2 1.2～ 10 6 .0μg/ml和 2 0 .0～ 10 0 .0μg/ml范围内 ,槲皮素和山奈素均呈良好的线性关系和重现性。 40 ,6 0和 80μl槲皮素的加样回收率分别为 93.87% ,94.0 2 %和 94.10 % ;40 ,6 0和 80μl山奈素的加样回收率分别为 94.5 0 % ,94.17%和 94.2 5 %。结论 :SFE与 MECC法联用 ,简便快速 ,可用于银杏叶粗提物中的黄酮类成分的分离分析。     

超临界CO_2萃取牛蒡片挥发油及气相色谱-质谱联用分析

目的:探讨超临界CO2萃取牛蒡片挥发油的可行性并检测其化学成分。方法:采用超临界CO2萃取得到牛蒡片挥发油,并用气相色谱-质谱联用(GC-MS)对其化学成分进行分析。结果:挥发油的萃取率为0.84%,从挥发油中分离出41个峰,鉴定出其中31种化合物。结论:超临界CO2萃取可作为一种有效提取牛蒡片挥发油的方法,其挥发油为多种类型化合物的混合物。     

藏药毕毕林二氧化碳超临界流体萃取精油化学成分研究

目的 研究习称为“三辛药”之一的藏药毕毕林精油化学成分及其相对含量.方法 采用超临界CO2流体萃取法提取毕毕林精油,采用气相色谱-质谱联用方法分离鉴定精油化学成分并测定其相对含量.结果 毕毕林超临界CO2流体萃取精油得率为9.7％,分离得到59个色谱峰,鉴定出52种成分,主要成分是胡椒碱(19.24％)、毕澄茄烯(l0...     

白藜芦醇与半边旗二萜类化合物5F对活体血管生成的影响

目的:观察白藜芦醇(Res)、半边旗二萜类化合物5F(以下简称5F)对活体血管生成的影响。方法:采用鸡胚绒毛尿囊膜(CAM)试验,检测Res和5F对活体血管生成的影响。结果:与对照组相比,Res用药组(11.4μg/egg、45.7μg/egg)的血管生成抑制率分别为(32.2±3.6)%、(53.7±4.2)%,5F用药组(8.3μg/egg、33.2μg/egg)血管生成抑制率分别为(19.7±4.0)%、(36.5±3.0)%。结论:Res和5F可抑制活体血管生成,对肿瘤和其他病理情况下的抗血管治疗可能具有重要应用价值。     

固相萃取-高效液相色谱法测定拟缺香茶菜中3个二萜类成分的含量

目的建立测定拟缺香茶菜药材中二萜类成分Kamebacetal-A、Kamebacetal-B、肾形香茶菜丙素的方法。方法采用固相萃取-高效液相色谱法(SPE-HPLC)测定3个二萜类成分的含量。使用Venusil C18(4.6 mm×250 mm,5μm)反相色谱柱;流动相:乙腈(A)-水(B)梯度洗脱;流速:1 m L/min;柱温:30℃;检测波长230 nm。结果拟缺香茶菜中Kamebacetal-A、Kamebacetal-B、肾形香茶菜丙素分别在0.056~0.448 g/L(r=0.999 2),0.002 6~0.020 8 g/L(r=0.999 4),0.016~0.128 g/L(r=0.999 7)内与峰面积成良好的线性关系;平均回收率分别为97.53%(RSD=2.16%),96.02%(RSD=2.01%),98.23%(RSD=2.10%)。结论所建立的方法灵敏、准确、稳定,可用于拟缺香茶菜中Kamebacetal-A、Kamebacetal-B、肾形香茶菜丙素的含量测定,为拟缺香茶菜抗肿瘤谱效关系的研究奠定基础。     

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.