一种快速微量外周血基因组DNA的提取方法

目的建立一种简便、快速、有效、微量外周血基因组DNA的提取方法.方法采用NaI作细胞裂解剂裂解细胞和细胞核,用氯仿:异戊醇(24:1)直接提取基因组DNA.结果该方法提取外周血基因组DNA的纯度高A260 nm/A280 nm=(1.87±0.11),提取效率每毫升外周血可得DNA(31±3.12)μg.结论该法简便、快速、有效,适应于批量临床标本的处理.     

全血基因组DNA快速提取法

目的　建立一种简便、快速、有效、微量外周血基因组DNA的提取方法。方法　采用NaI作细胞裂解剂裂解细胞和细胞核 ,用氯仿∶异戊醇 ( 2 4∶1 )直接提取基因组DNA。结果　该方法提取外周血基因组DNA的纯度高 ,A2 60nm/A2 80nm=1 .83± 0 .1 3,每毫升外周血可得DNA 33± 3.5 2 μg。结论　本法简便、快速、有效 ,适用于批量临床标本的处理。     

人类外周血基因组DNA提取方法的比较分析

目的:对2种提取外周血基因组DNA的实验方法进行了比较,建立一种本实验室有效的外周血基因组DNA的提取方法.方法:采用常规试剂盒和本实验室设计的2种外周血液DNA提取法.结果:本实验室使用的外周血基因组DNA提取方法与试剂盒提取外周血基因组DNA的纯度比较无明显差异.结论:本实验室使用的外周血液DNA提取方法简便、有效、经济,满足于基因组的研究.     

外周血DNA提取方法的比较及改良

目的 对3种提取外周血的实验方法进行了比较，并建立一种简便、快速、有效的外周血基因组DNA的提取方法。方法 采用常规酚／氯仿提取法、碘化钾法和外周血液DNA提取法。结果 外周血液DNA提取方法提取外周血基因组DNA的纯度高，通过电泳图明显可见。结论 外周血液DNA提取简便、快速、有效适用于临床标本的研究。     

煮沸裂解法快速提取大鲵DNA

目的建立大鲵基因组DNA快速简便高效的提取方法。方法借鉴煮沸裂解法制备质粒DNA方法,改进后用于提取大鲵基因组DNA。结果本方法提取DNA纯度高（A260/A28介于1.7~2.1之间）,耗时短（2h）,毒性小,分子大小约为23kb,适用于PCR扩增及后续分析。结论本方法提取基因组DNA纯度高、耗时短、提取所用试剂毒性小、成本低,值得在大鲵基因组DNA提取中使用,也可为其他动物的相关研究提供参考。     

血液基因组DNA提取方法的改良简

目的 探讨改良血液样品基因组DNA的提取方法,制备纯化的高分子量DNA。方法 将312份新鲜血液样品随机分为2组:A组(90份)和B组(222份)。A组采用常规基因组DNA提取方法。B组采用改良的基因组DNA提取方法,其基因组DNA提取方法改良之处:1)在血液样品中加入细胞裂解液CL后采用手工震荡,并离心2min 40s;2)震荡混匀后将样品分装成3管,然后将缓冲液GS加入第1管吹打溶解后吸取上清,并加入第2管,再吹打溶解后吸取上清加入第3管,继续吹打溶解;3)加入缓冲液GB后再58℃水浴过夜16h。结果 改良的血液样品DNA提取方法能够提高产率,提高纯度,获取更多高分子量的DNA,相对于常规基因组DNA提取方法有显著提高。结论 改良的血液基因组DNA提取方法操作简便,能有效地避免操作导致DNA链断裂,提高DNA的完整性及纯度。     

一管酒精法提取转基因小鼠胚胎组织基因组的实验方法及应用

目的:优化提取基因组的方法,建立高效、简便、快速提取基因组DNA的方法,用于大批量转基因小鼠鉴定。方法:用一管酒精基因组DNA抽提法抽提基因组DNA。结果:经过分光光度法检测基因组DNA浓度、纯度及电泳分析DNA质量,显示一管酒精抽提法提取的基因组与经典酚/氯仿提取法提取的基因组产量、纯度及质量没有明显差别;酶切全基因组后,2种方法获得的基因组都可以被酶完全切开;两者模板都能成功应用PCR扩增来鉴定基因敲入小鼠并可应用于基因组测序及Real-time PCR检测基因敲入小鼠中外源基因并鉴定其相对拷贝数。结论:一管酒精基因组抽提法得到的DNA质量可靠,可高效、简便、快速鉴定转基因小鼠及进行基因组DNA酶切、测序、Real-time PCR等后续实验。     

一种从培养细胞中快速制备病毒PCR模板的方法

目的建立一种快速、简单的从培养细胞中制备病毒PCR模板的方法。方法分别用裂解液煮沸法、苯酚法及基因组DNA提取试剂盒提取F81细胞中的犬细小病毒的DNA,分析3种方法提取的病毒DNA纯度、浓度及操作所需时间,并作巢式PCR检测,比较其PCR检测的阳性率。结果以裂解液煮沸法提取的犬细小病毒DNA为模板进行巢式PCR检测后的阳性率与两种对照方法之间的差异无统计学意义(P>0.05),3种方法提取的病毒DNA纯度及浓度未见差异,但裂解液煮沸法操作所需时间最短。结论裂解液煮沸法是一种快速而简单的从培养细胞中制备病毒PCR模板的方法。     

混合菌基因组的合并提取及鉴定

目的为了简便、快速的提取混合病原菌的基因组DNA，进而获得不同病原菌的特异性毒力DNA片断用于进一步的基因诊断。方法改良了裂解试剂，同时提取革兰氏阳性菌(以金黄色葡萄球菌为代表)和革兰氏阴性菌(以产毒性大肠杆菌ETEC为代表)的混合基因组DNA；采用双重PCR法进行扩增鉴定。结果提取得到混合菌的混合基因组，并以此为模板通过双重PCR同时扩增得到两种菌的特异性DNA片断。结论这是一种合并提取混合菌基因组DNA，进而获得不同病原菌DNA片段的简便方法。     

一种从培养细胞中快速制备病毒PCR模板的方法

目的建立一种快速、简单的从培养细胞中制备病毒PCR模板的方法。方法分别用裂解液煮沸法、苯酚法及基因组DNA提取试剂盒提取F81细胞中的犬细小病毒的DNA,分析3种方法提取的病毒DNA纯度、浓度及操作所需时间,并作巢式PCR检测,比较其PCR检测的阳性率。结果以裂解液煮沸法提取的犬细小病毒DNA为模板进行巢式PCR检测后的阳性率与两种对照方法之间的差异无统计学意义（P〉0.05）,3种方法提取的病毒DNA纯度及浓度未见差异,但裂解液煮沸法操作所需时间最短。结论裂解液煮沸法是一种快速而简单的从培养细胞中制备病毒PCR模板的方法。     

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.     

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.