中药材中八种有机磷类农药残留的检测方法研究

目的建立中药材中八种有机磷类农药残留的检测方法,为中药材安全使用标准的建立提供科学依据。方法八种农药残留的样品以丙酮超声提取,并采用凝胶渗透色谱(GPC)和石墨碳柱结合的方法进行净化,选用气相色谱-质谱串联技术(GC/MS)在DB-17MS毛细管柱上用程序升温技术分离和离子检测方式,以外标法对样品进行测定。结果八种农药可在24 min内完全分离,工作曲线在10μg/L～1000μg/L的范围内有良好的线性关系,相关系数在0.9991～0.9999的范围内,检测限为0.6μg/kg～2.3μg/kg,三个水平添加回收率在71%～109%之间,相对标准偏差小于10%(n=9)。结论本方法灵敏度高,重现性好,适用于中药材中八种农药的残留检测。     

超高效液相色谱-串联质谱法测定面粉、玉米及其制品中的伏马菌素研究

目的建立面粉、玉米及其制品中3种伏马菌素的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。方法样品用乙腈-水(50∶50,V/V)溶液提取,经MultiSep 211净化柱富集净化后,用甲醇-甲酸(99∶1,V/V)溶液洗脱,洗脱液用UPLC-MS/MS进行测定。UPLC-MS/MS检测伏马菌素的条件为梯度洗脱,流动相为0.2%甲酸水和乙腈;质谱采集模式为电喷雾正离子多反应监测模式。结果 UPLC-MS/MS检测方法对3种伏马菌素的检出限均为0.2μg/kg,最低定量限为0.6μg/kg。不同加标水平的3种伏马菌素平均回收率为69.44%～114.69%。用所建方法对2011年甘肃及天津生产的39份玉米及30份面粉样品进行了分析,结果以伏马菌素B1的检出率最高,污染水平最为严重。结论 UPLC-MS/MS法具有操作简单、灵敏度高、重现性好等特点,可用于面粉、玉米及其制品中伏马菌素的检测。     

雾霾PM2.5中7种指示性多氯联苯的微波辅助萃取气相色谱三重四极杆串联质谱法

目的建立雾霾PM2.5中痕量7种指示性多氯联苯的微波辅助萃取气相色谱三重四极杆串联质谱(GC-MS/MS)检测方法。方法采样后的雾霾PM2.5玻璃纤维滤膜经微波辅助萃取,佛罗里硅土柱净化后,用GC-MS/MS测定。结果 7种指示性多氯联苯在0.5~100μg/L范围内线性良好,线性回归系数r≥0.999,仪器检出限为0.03~0.06μg/L,方法检出限为0.042~0.084 pg/m~3,加标回收率为74.5%~94.5%,相对标准偏差RSD(n=6)为2.1%~3.6%。结论该方法灵敏、准确,有效去除了基质干扰,适用于雾霾PM2.5中7种指示性多氯联苯的测定。     

超高效液相色谱法检测食品黄曲霉毒素

目的 建立一种快速、灵敏、简便的超高效液相色谱法(UPLC)同时测定食品中黄曲霉毒素B1(AFB1)、B2(AFB2)、G1(AFG1)、G2(AFG2)水平的方法.方法 样品用乙腈水(84:16)提取液提取后过滤,滤液经黄曲霉毒素多功能净化柱(MFC)净化,纯化液经浓缩挥干后,加三氟乙酸(TFA)衍生后,用带有荧光检测器的超高效液相色谱仪测定.结果 4种黄曲霉毒素5min完全分离,线性范围质量浓度分别为0.10～100.00μg/L(AFB1、AFG1)、0.05～50.00μg/L(AFB2、AFG2),最低检出质量分数分别为0.04μg/kg(AFB1、AFG1)、0.02μg/kg(AFB2、AFG2),大米、玉米、花生、油炸面点回收率为80.0%～101.2%,相对标准偏差＜5%,r≥0.999.结论 该方法简单、快速、灵敏度高,适用于食品中4种黄曲霉毒素水平的同时测定.     

气相色谱串联质谱法测定饮用水中多种农药及其代谢产物残留

目的建立液液萃取-气相色谱串联质谱(Liquid liquid extraction and gas chromatography-tandem mass spectrometry,LLE-GC-MS/MS)测定饮用水中28种农药残留和代谢产物的检测方法。方法水样用二氯甲烷:正己烷(1∶1,V/V)溶剂提取后,合并提取液用氮气浓缩至1 ml,采用气相色谱串联质谱(gas chromatography-tandem mass spectrometry,GC-MS/MS)的多反应监测模式(multi reaction monitoring,MRM)对多种农药及其代谢产物残留同时定性定量分析,采用DB-5MS色谱柱(30 m×0.25 mm,0.25μm)分离。结果 28种农药在0.1~4.0 mg/L线性范围内均呈较好的线性关系,r0.998。该方法的检出限为0.018~0.066μg/L。在1.6μg/L和4.0μg/L两种浓度添加水平,其平均回收率为85.3%~101.8%,相对标准偏差(relative standard deviations,RSD)为3.8%~9.2%。结论该方法快速、简单、准确,适用于同时检测饮用水中多种农药残留和代谢产物残留。     

免疫亲和柱净化-碘柱后衍生化-高效液相色谱荧光法检测斑蝥中黄曲霉毒素及其液质确证

目的:建立高效液相色谱-碘柱后衍生化-荧光检测器检测中药材斑蝥中黄曲霉毒素B1、B2、G1、G2的含量,并采用液质联用法进行确证。方法:样品以70%甲醇溶液提取,经免疫亲和柱净化后,利用碘柱后衍生化-HPLC-FLD进行分析测定。并进一步采用LC-MS/MS对阳性样品进行确证。结果:在优化条件下,黄曲霉毒素B1、B2、G1、G2分别在0~0.525、0~0.1125、0~0.1875、0~0.1125μg的范围内线性关系良好,r0.9996,回收率82.5%~107.3%,RSD8.1%。检测了21批样品并通过LC-MS/MS确证,在与对照品相同的保留时间处,样品与对照品有相同的特征离子碎片,排除了样品假阳性的可能。结论:该方法简便快速,灵敏度高,重复性好,适用于斑蝥中黄曲霉毒素的检测。     

高效液相色谱法测定粮食中伏马菌素B1

目的建立伏马菌素B(1FB1)的高效液相色谱检测方法;了解辽宁地区玉米产品中FB1的污染状况。方法利用强离子交换固相提取柱(SPE柱)净化样品提取液,选用C18反相柱,以甲醇∶NaH2PO4为流动相,流速1ml/min;荧光检测器检测。结果1μg/ml、5μg/ml、10μg/ml3个加标浓度的回收率在71.77%～102.68%之间,平均回收率为84.72%,总变异系数为12.17%。检测78份玉米样品,阳性率为32.05%,阳性样品中FB1平均含量为(0.74±0.05)μg/g。结论建立了用高效液相色谱法检测FB1的方法;本实验检测结果提示辽宁地区玉米样品的FB1污染率较低。     

ELISA法测定贝毒素污染的研究

目的 探讨以ELISA检测麻痹性贝毒素(PSP)、腹泻性贝毒素(DSP)、神经性贝毒素(NSP)的可行性,并对深圳市污染状况调查.方法 采用ELISA检测PSP、DSP、NSP,对抽样的深圳市贝类样品进行检测.结果 PSP浓度为0.02～0.32μg/kg之间的标准曲线R=0.996,变异系数(CV,％)为2.4～8.3,回收率为88.7％～105.0％; DSP浓度为0.20～5.00μg/kg之间的标准曲线R=0.995,变异系数(CV,％)为1.5～8.0,回收率为89.6％～116.0％; NSP浓度为0.10～2.50μg/kg之间的标准曲线R=0.993,变异系数(CV,％)为2.2～9.5,回收率为88.7％～111.6％.结论 ELISA法检测三种贝毒均具有灵敏度高、干扰少、测定步骤简便、快速、操作安全、结果准确可靠的优点,适合作为快速筛选方法.     

食品中黄曲霉毒素测定方法的优化研究

目的建立高效液相色谱等浓度流动相同时测定黄曲霉毒素G_1、B_1、G_2、B_2的方法。方法样品经乙腈+水(84+16)溶解,用多功能振荡器超声提取30 min,多功能固相萃取柱净化,将净化液收集于10 ml比色管中,取少量净化液于10 ml一次性离心杯中,60℃水浴氮气吹至近干,用正己烷和三氟乙酸在40℃培养箱内衍生15 min,室温干燥,用乙腈+甲醇+水(22+22+56)溶解,过0.22μm小滤膜于样品瓶中,荧光检测器,流动相乙腈+甲醇+水(22+22+56)测定。结果黄曲霉毒素G_1、B_1、G_2、B_2分离效果好,回收率在83.7%~96.5%之间,RSD值在0.92%~2.74%之间,线性关系r=0.9 991~0.9 993,检出限分别为:0.050μg/kg,0.025μg/kg,0.100μg/kg,0.030μg/kg。结论高效液相色谱法优化测定食品中黄曲霉毒素灵敏度好,色谱条件比GB/T 5009.23-2006的梯度洗脱简单,快速,定量准确。     

气相色谱三重四级杆串联质谱法测定雾霾PM2.5中12种共平面多氯联苯

目的建立雾霾PM2.5中12种共平面多氯联苯的前处理及气相色谱三重四级杆串联质谱(gas chromatography tandem mass spectrometry,GC-MS/MS)分析方法。方法石家庄市采集的雾霾PM2.5颗粒物样品,用正己烷丙酮混合溶剂经加速溶剂萃取仪(accelerated solvent extraction,ASE)萃取,佛罗里硅土柱净化,氮吹后正己烷定容,气相色谱三重四级杆串联质谱多重反应监测(multiple reaction monitoring,MRM)模式测定,外标法定量。3倍信噪比确定检出限。结果 12种共平面多氯联苯在1~100μg/L范围内线性良好,相关系数R~20.999。该方法的仪器检出限为0.034~0.11μg/L,方法检出限为0.05~0.16 pg/m~3。3个水平的加标回收率为77.0%~110.2%,相对标准偏差(RSD)在2.8%~11.2%之间。结论净化步骤和优化的仪器参数使目标物能够准确地检测和定量,该方法简便快速、溶剂用量少、基质干扰少,对客观表达雾霾PM2.5中污染状态信息有重要意义,适用于雾霾PM2.5中痕量共平面多氯联苯的检测要求。     

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.