实时荧光定量聚合酶链反应快速检测下呼吸道痰液标本耐甲氧西林金黄色葡萄球菌的临床应用

目的 应用实时荧光定量聚合酶链反应(PCR)检测耐甲氧西林金黄色葡萄球菌(MRSA)核酸,寻求呼吸道标本MRSA的快速检测技术.方法 收集临床分离的75株金黄色葡萄球菌菌株及97份下呼吸道感染患者的合格痰液标本,应用实时荧光定量PCR快速检测法检测标本中Nuc及MecA基因,并以常规培养+头孢西丁纸片扩散法的检测结果为标准,判断实时荧光定量PCR与传统方法检测结果的一致性,及该方法对下呼吸道痰液标本检测的敏感度、特异度.结果 以常规培养+头孢西丁纸片扩散法的检测结果为对照,实时荧光定量PCR快速检测法检测金黄色葡萄球菌菌株中MRSA的敏感度及特异度均为100.0％,检测下呼吸道痰液标本MRSA的敏感度为100.0％、特异度为84.7％.结论 应用实时荧光定量PCR快速检测下呼吸道痰液标本MRSA的敏感度高,特异性强,操作时间短,与常规培养+药物敏感试验的检测结果具有良好的一致性,具有较高的临床应用价值.     

异基因造血干细胞移植受者CMV检测方法的比较

目的:比较异基因造血干细胞移植(allo-HSCT)后早期有效检测巨细胞病毒(CMV)激活感染的方法。方法:分别应用荧光定量PCR和ELISA试剂盒检测57名allo-HSCT受者642份标本的血浆DNA负荷量和血清IgM抗体。结果:IgM抗体和DNA定量的阳性检出率为35.51%(228/642)和13.08%(84/642),连续阳性病例和临床诊断的符合率分别为33.33%(9/24)和87.5%(21/24),IgM抗体阳性检出率明显低于DNA定量,差异有统计学意义(P<0.05)。结论:荧光定量PCR检测allo-HSCT受者CMV早期感染可靠、简便快速,值得临床推广使用。     

实时荧光定量PCR法在急性呼吸道传染病病原筛查中的应用

目的利用实时荧光定量PCR法筛查和鉴定某战区发热伴呼吸道症状病例的呼吸道病毒感染情况,为部队呼吸道传染病疫情的早期预警和防控提供科学依据。方法采集某战区发热伴呼吸道症状病例咽拭子标本,采用实时荧光定量PCR法对标本进行流感病毒、副流感病毒、腺病毒、呼吸道合胞病毒和冠状病毒5类常见呼吸道病毒及其亚型的检测和鉴定,评价该方法在呼吸道病毒检测中的应用价值。结果利用实时荧光定量PCR法在采集的488份咽拭子标本中筛查出了240份甲型流感病毒阳性标本(其中季节性流感H3亚型71份,其它分型未检出)、7份乙型流感病毒阳性标本,未检出副流感病毒、腺病毒、呼吸道合胞病毒和冠状病毒。结论实时荧光定量PCR法可以对呼吸道病毒进行快速、特异、准确的筛查和鉴定,科学指导部队对呼吸道传染病疫情进行早期防控和预警预测。     

多重实时荧光定量PCR检测急性肝炎患者HBV和HCV方法的建立与意义分析

目的建立一种新的可以快速筛查急性肝炎患者HBV和HCV感染的多重实时荧光定量PCR方法。方法收集596例急性肝炎患者血清或血浆样本,应用自主构建的多重实时荧光定量PCR方法同时检测HBV和HCV,同时应用HBV和HCV单管检测试剂进行检测分析。结果新建立的多重实时荧光定量PCR体系,可以有效地检测HBV和HCV的阳性质控品及阳性样本。多重实时荧光定量PCR体系特异性较好,检测HBV和HCV时,两者之间无交叉反应,且以多种血浆病毒的核酸样品为模板进行检测时结果全为阴性;多重实时荧光定量PCR体系检测HBV、HCV的灵敏度分别为20 IU/ml、50 IU/ml;多重实时荧光定量PCR体系精密度好,检测HBV、HCV的CV值分别为0.11%、0.096%。在检测临床样本时,多重复合体系检测血清或血浆中HBV和HCV的检出率与单独检测HBV和HCV具有很好的一致性,且HBV和HCV的一致率分别达到97.7%(k=0.952)、98.8%(k=0.976);同时有6例样本出现HBV和HCV共感染。结论新型多重实时荧光定量PCR方法对快速检测及筛查HBV和HCV较快捷,而且特异性好、灵敏度高、精密度好。因此多重实时荧光定量PCR方法对2种肝炎病毒的检测具有重要意义。     

MP DNA检测对小儿支原体肺炎的诊断价值

目的探讨肺炎支原体（MP）DNA检测在小儿支原体肺炎早期诊断中的价值。方法用荧光定量聚合酶链反应（FQ—PCR）技术对我院2009年1-11月805例疑似MP感染患儿的咽拭子标本进行MPDNA检测,同时采取静脉血检测805例患儿血清中MP-IgM抗体。结果805倒患儿咽拭子标本MPDNA检测阳性率显著高于血清中MP-IgM抗体检测阳性（P〈0.05）。结论PCR技术可以快速、敏感、准确地定量检测标本中MPDNA,对儿童MP感染的早期、快速诊断诊断具有重要的参考价值。     

定量PCR结合混合标本检测献血者HCV-RNA

目的:探讨应用核酸扩增技术(NAT)定量检测献血者混合标本方法的可行性。方法:用荧光定量PCR法检测38例抗-HCV阳性标本、3 274例抗-HCV阴性标本及二者随机抽样组成的混合标本的HCV-RNA含量。结果:在38例抗-HCV阳性标本中有15例核酸检测阳性,两者阳性符合率39.5%(15/38)。15例双阳性标本与抗-HCV阴性标本混合检测核酸,仍为阳性。常规血清学检测阴性的标本NAT检测为阴性。结论:定量PCR检测HCV-RNA的方法适用于献血者的筛查。     

FQ-PCR同步检测HTLV-Ⅰ及HTLV-Ⅱ方法的建立及临床应用

目的:建立可同时检测HTLV-Ⅰ和HTLV-Ⅱ的双重荧光定量PCR(FQ-PCR)方法,并对所建体系进行方法学评价.方法:根据HTLV-Ⅰ pol基因和HTLV-Ⅱtax基因的保守序列设计引物及TaqMan-LNA探针,构建重组质粒作为荧光定量PCR的标准品.优化荧光定量PCR反应条件,建立起可同时检测HTLV-Ⅰ和HTLV-Ⅱ的双重荧光定量PCR (FQ-PCR)的方法.结果:结果显示该方法对HTLV-Ⅰ及HTLV-Ⅱ的线性范围分别为108-102 copy/μl和107-10 copy/μl;检测灵敏度分别为102copy/μl和10 copy/μl.对阳性标本检测均呈阳性,对非目标菌检测均呈阴性.应用该方法对175例临床标本进行考核,有HTLV-Ⅰ阳性6例,HTLV-Ⅱ阳性3例,其中HTLV-Ⅰ和HTLV-Ⅱ双阳性2例.结论:本研究建立的双重FQ-PCR方法特异、快速、灵敏,对HTLV的血液筛查有重要意义.     

肾移植受者巨细胞病毒隐性感染的筛查方法及应用

目的探讨肾移植受者巨细胞病毒(cytomegalovirus,CMV)隐性感染的筛查方法,为临床早期发现CMV活动性感染提供诊断依据。方法收集笔者医院2009年10～12月期间门诊随访233例肾移植受者外周静脉血标本450份,分别采用实时荧光定量PCR(Real-time PCR)、巢式PCR(nPCR)以及酶联免疫吸附(ELISA)法对标本进行检测,同时采用ELISA法检测他克莫司(Tac)浓度。结果 233例肾移植受者CMV-IgG均为阳性,CMV-IgM则均为阴性;Real-time PCR技术检测CMV-DNA阳性85例(36.48%),以低拷贝数为主。nPCR技术检测CMV-DNA阳性10例(4.29%),两种技术检测总阳性率存在显著性差异(P<0.05);随着Tac药物浓度升高,CMV-DNA阳性率呈增高趋势,药物高浓度组CMV-DNA阳性率高于低浓度组(P<0.05)。结论 Real-time PCR是筛查肾移植受者CMV隐性感染较为理想的方法,尤其在受者药物浓度偏高时更具有临床应用价值。     

酶联免疫吸附和胶体金与荧光定量PCR技术在乙肝病毒检测中的应用

目的探讨酶联免疫吸附(ELSIA)、胶体金与荧光定量PCR检测方法在乙肝病毒检测中的应用价值。方法采用ELSIA分析法及快速胶体金方法分别对标准品血清、阴阳性对照血清及43份随机抽样血清进行乙肝两对半检测;对乙肝两对半中不同项目阳性的68份血清标本做荧光定量PCR(FQ-PCR)检测,并将检测结果进行对比分析。结果ELSIA、胶体金两种方法检测结果比较,HBsAg、HBsAb阳性符合率达90.2%,HBeAg阳性符合率达到100%,HBeAb、HBcAb阳性符合率分别为80.1%、77.8%。HBsAg、HBeAg及HBcAb均阳性者荧光定量PCR阳性率达93.33%。结论ELSIA方法具有较好的灵敏度、特异性。胶体金检测方法可以避免Hock效应所致的假阴性结果,可作为急诊筛查检测方法。HBeAg阳性与HBVDNA阳性吻合度最高,是判定病毒复制与传染性的可靠指标。     

荧光定量PCR和超高倍显微诊断系统检测沙眼衣原体

目的探讨荧光定量PCR和超高倍显微分析仪活体检测泌尿生殖道沙眼衣原体感染的诊断价值。方法应用荧光定量PCR和MMDI法对40例患者进行沙眼衣原体(CT)检测,并将两种方法进行比较。结果两者均阳性18例(45.0%);两者阴性19例(47.5%),荧光定量PCR阳性、MMDI阴性2例(5.0%);荧光定量PCR阴性、MMDI阳性1例(2.5%)。两法检出率比较,差异无统计学意义(McNewmar检验,P=1.000)。结论实验证明荧光定量PCR和MMDI均能很好地检测沙眼衣原体。MMDI是检测沙眼衣原体的一种快速、直观的诊断方法,而荧光定量PCR检测较为客观。     

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.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.     

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.