PCR和分离培养检测细胞培养支原体污染的比较

目的：检测细胞培养中支原体的污染,建立支原体通用的ＰＣＲ方法。方法：根据已出版的序列设计并合成一对支原体（柔膜体纲）１６Ｓ ｒＲＮＡ基因组特异引物。对实验室所存的１２种支原体、大肠杆菌、变形杆菌及无支原体污染的组织培养细胞进行了ＰＣＲ扩增。以ＰＣＲ与分离培养比较检测了３３份细胞培养标本。结果：１２种支原体均出现了约６２０ｂｐ左右的特异性扩增带,敏感性为１００￣１０００个支原体细胞,且常见的６种细胞     

16SrRNA基因PCR对新生儿败血症的诊断研究

目的：探索快速可靠的新生儿败血症诊断的新方法。方法：通过自行设计合成细菌１６ＳｒＲＮＡ基因高度保守区引物，对２０种标准菌株、１２种２７株临床分离细菌基因组ＤＮＡ及巨细胞病毒进行ＰＣＲ扩增。结果：细菌具有３７１ｂｐ扩增产物，与人基因组ＤＮＡ、巨细胞病毒无交叉阳性反应，ＰＣＲ最低能检测１ｐｇ大肠杆菌ＤＮＡ。结论：建立了用共同引物ＰＣＲ扩增诊断败血症病原菌的方法，检测快速，特异性、敏感性高。     

用双重PCR快速诊断新生儿巨细胞病毒感染

用双重ＰＣＲ法检测新生患儿及随访新生儿期诊断为巨细胞包涵体病患儿的尿及血标本中巨细胞病毒（ＣＭＶ）ＤＮＡ。两对引物序列取自ＨＣＭＶＩＥＡ基因区，第一对引物扩增片段为７２１ｂｐ，第二对引物扩增片段为１６７ｂｐ。新生患儿２３份尿标本，１７例检出阳性；１８份白细胞提取物，１２份阳性。随访患儿７例，４例在尿或血中检出ＣＭＶＤＮＡ。ＰＣＲ结果与病毒分离比较，两法的符合率为９５．６％，ＰＣＲ获得结果仅需１～２天。尿标本用于ＰＣＲ较血标本检出率高，预处理简便     

16 SrRNA基因聚合酶链反应检测细菌感染的研究

目的：探索快速可靠的检测细菌感染的新方法。方法：通过自行设计合成的细菌１６ＳｒＲＮＡ基因高度保守区引物,对２０ 种标准菌株、１２ 种２７ 株临床分离的细菌株、人基因组ＤＮＡ及巨细胞病毒进行聚合酶链反应（ＰＣＲ）扩增。结果：对所测细菌株均获得３７１ ｂｐ 扩增产物,而与人基因组ＤＮＡ、巨细胞病毒无交叉阳性反应,ＰＣＲ最低能检测ｌｐｇ 大肠杆菌ＤＮＡ。结论：建立了用共同引物ＰＣＲ扩增以判断是否存在细菌感染的方法,该方法检测快速,敏感性和特异性高。     

采用通用引物进行PCR检测宫颈中人乳头瘤病毒的基因

采用可检测九个型别人乳头瘤病毒（ＨＰＶ）的通用引物进行多聚酶链反应，在４７份宫颈糜烂患者的宫颈细胞标本中检出了ＨＰＶ阳性的标本２３份。在这２３份阳性标本中ＨＰＶ１６型特异性引物进一步用ＰＣＲ进行扩增，检出ＨＰＶ１６型阳性者１０份，在２４份ＨＰＶ阴性的标本中用ＨＰＶ１６型特异性引物未检出ＨＰＶ１６阳性的标本。结果表明：用通用引物进行ＰＣＲ检测人乳头瘤病毒的感染，方法简便、灵敏、经济，特别适合于ＰＣＲ     

嵌套式PCR法分型检测登革病毒基因

应用一对通用引物同时扩增４个型登革病毒（ＤＥＮ）的ＮＳ１基因部分片段，然后采用嵌套式聚合酶链反应（ＰＣＲ）方法对ＤＥＮ进行分型鉴定。通用引物扩增基因序列长度为４１３ｂｐ；ＤＥＮ各型内引物扩增序列长度分别是：ＤＥＮ１型为２６２ｂｐ；ＤＥＮ２型为１８９ｂｐ；ＤＥＮ３型为３９２ｂｐ；ＤＥＮ４型为９７ｂｐ。应用嵌套式ＰＣＲ法直接分型检测登革热患者血清标本２１份，发现ＤＥＮ１型阳性１８份，ＤＥＮ２型阳性２份。该法能在２ｄ内完成对ＤＥＮ的分型鉴定，为登革热的早期快速诊断提供了可靠的方法。     

日本血吸虫中国大陆株副肌球蛋白编码基因的PCR扩增

根据已知的日本血吸虫菲律宾株的副肌球蛋白分子的部分ｃＤＮＡ序列，设计两对寡核苷酸引物（引物１／２和引物３／４），以聚合酶链式反应（ＰＣＲ），用引物１／２从本室两个日本血吸虫中国大陆株ｃＤＮＡ库中均扩增出与预期大小（９２７ｂｐ）一致的特定ＤＮＡ片段。巢式ＰＣＲ——以第一扩增产物为模板，用引物３／４扩增出约５００ｂｐ的单一条带，与预期片段（４９４ｂｐ）大小一致。表明ＰＣＲ产物为编码副肌球蛋白的目的基因片段。     

日本血吸虫大国大陆株副肌球蛋白编码基因的PCR扩增

根据已知的日本血吸虫菲律宾株的副肌球蛋白分子的部分ｃＤＮＡ序列，设计两对寡核苷酸引物（引物１／２和引物３／４），以聚合酶链式反应（ＰＣＲ），用引物１／２从本室两个日本血吸虫中国大陆株ｃＤＮＡ库中均扩增出与预期大小（９２７ｂｐ）一致的特定ＤＮＡ片断。巢式ＰＣＲ－以第一扩增产物为模板，用引物３／４扩增出约５０００ｂｐ的单一条带，与预期片段（４９４ｂｐ）大小一致。表明ＰＣＲ产物为编码副肌球蛋白的目的基因     

立克次体感染的PCR检测

作者应用ＰＣＲ技术对３０例疑似立克次体感染或不明热患者的血液标本和／或立克次体细胞分离培养物进行立克次体特异性ＤＮＡ检测。用Ｒｒ．１９０．７０ｐ、Ｒ．ｒ．１９０６０２ｎ，Ｒｒ．１７ＫＤ，ｓｔａ５８３种引物对分别进行ＤＮＡ扩增，共检出斑点热群立克次体特异性ＤＮＡ１１份，斑疹伤寒立克次体特异性ＤＮＡ４份，恙虫病立克次体特异性ＤＮＡ４份，其中有１０份血液标本在分离前用Ｒｐ．Ｃ．Ｓ．引物进行ＰＣＲ检测，检出立克次体属ＤＮＡ７份，分离培养后再取分离培养物和阳性血液标本同法用Ｒｒ．１９０．７０ｐ、Ｒ．ｒ．１９０６０２ｎ，Ｒｒ．１７ＫＤ，ｓｔａ５８３种引物对分别进行ＰＣＲ检测，分离前后所得结果一致     

幽门螺杆菌细胞空泡毒素的PCR检测

目的：建立幽门螺杆菌（ＨＰ）细胞空泡毒素（ＶＣ）的聚合酶链反应（ＰＣＲ）检测方法。方法：根据ＨＰＶＣ基因图谱，设计了两对顺序特异性引物，分别扩增包括信号肽顺序（４０６－１５１６）和不含此顺序（８１１－１５１６）的基因片段。对１４株经形态学和生化鉴定证实为ＨＰ的菌株提取其基因组ＤＮＡ，以上述引物进行ＰＣＲ。结果：已知ＶＣ阳性的ＨＰ标准株（ＮＣＴＣ１１６３８）、临床分离株ＮＺ７、ＮＺ８、ＨＰ２３、ＨＰ３５、ＨＰ５７、ＨＰ６５、ＨＰ６６、ＨＰ６７和ＨＰ７０共１０株为ＶＣ阳性。扩增产物中均检出ＶＣ特异的１１００ｂｐ、７０５ｂｐ、５４３ｂｐ片段，敏感性约为０．６ｎｇ基因组ＤＮＡ。结论：所设计的引物和ＰＣＲ检测方法正确，可应用于临床检测。     

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.