表皮葡萄球菌ica操纵子与细胞间多糖粘附素及生物膜表型的相关性

目的 探讨表皮葡萄球菌 ica 操纵子与细菌间多糖粘附索(polysaccharide intercellular adhesin,PIA)及生物膜表型之间的关系.方法 采用聚合酶链反应(PCR)检测 icaA 基因,刚果红琼脂平皿检测 PIA,微量板半定量法检测细菌生物膜表型;统计学分析 icaA 基因与 PIA 及生物膜表型之间的相关性.结果 49 株临床分离菌株中,32.7%(16/49)能形成生物膜,67.3%(33/49)无生物膜形成.icaA 基因阳性菌株中 80%(8/10)有生物膜形成,ica 操纵子与生物膜表型密切相关(P<0.01);PIA 阳性菌株中63.6%(14/22)有生物膜形成,PIA 与生物膜表型密切相关(P<0.01).结论 表皮葡萄球菌 ica 操纵子的存在与生物膜表型密切相关,PIA 的合成是生物膜形成的关键环节.     

ica操纵子对表皮葡萄球菌在不同骨科植入物表面黏附和生物膜形成的影响

目的 探讨ica操纵子对表皮葡萄球菌在骨科植入物表面黏附和生物膜形成能力的影响,为临床骨科材料的使用提供理论依据.方法 采用PCR扩增表皮葡萄球菌ica操纵子的icaADBC基因,荧光定量PCR检测ica操纵子基因的表达,苯酚-硫酸法检测细胞间多糖黏附素(PIA)的合成,黏附能力检测采用菌落平板计数法,生物膜形成能力检测采用结晶紫染色法.结果 11株表皮葡萄球菌临床菌株扩增出ica操纵子的icaADBC基因,且在菌株中均有表达.ica操纵子基因表达高的菌株其PIA分泌量较高.表皮葡萄球菌在骨组织上的黏附和生物膜形成能力最强,其次为钛合金和不锈钢.icaA基因表达与细菌在3种生物材料上的生物膜形成相关.结论 表皮葡萄球菌在不同骨科植入物上具有不同的黏附和生物膜形成能力,ica操纵子基因通过调节PIA分泌量而影响表皮葡萄球菌生物膜的形成.     

trap基因表达在表皮葡萄球菌生物膜形成及附属基因调节(agr)系统活化中的作用

目的研究表皮葡萄球菌trap基因表达与生物膜形成的相关性及其对agr系统的活化作用。方法采用微量板半定量法检测表皮葡萄球菌生物膜表型，二维电泳和高灵敏度的基质辅助激光解吸附飞行时间质谱分析比较表皮葡萄球菌标准株蛋白质表达谱，实时定量逆转录PCR检测trap基因和RNAⅢ的转录水平，CLUSTALX对表皮葡萄球菌trap基因与金黄色葡萄球菌序列进行分析，Mega软件构建进化树。结果比较分析形成生物膜的表皮葡萄球菌ATCC 35984株与不形成生物膜的ATCC 12228株生长中期的蛋白质组，发现ATCC 35984株的TRAP蛋白量明显高于ATCC 12228株。转录水平的检测显示ATCC 12228株和生物膜阳性临床SE1457、SE671株的trap基因均低转录，但RNAⅢ的转录水平未降低。SE1457株agr系统突变后，RNAⅢ的转录明显降低，trap基因的转录无变化。RNAⅢ的转录随细菌生长变化而增加，trap基因转录保持不变。trap序列分析显示其在表皮葡萄球菌与金黄色葡萄球菌中较为保守，但存在明显的进化距离。结论在本论文所研究的表皮葡萄球菌菌株中，trap基因的转录和翻译与细菌生物膜的形成不存在直接的调控相关性；并提示TRAP蛋白不是表皮葡萄球菌agr系统活化所必需的细菌产物；表皮葡萄球菌trap基因序列与金黄色葡萄球菌之间存在差异，可能与其功能差异相关。     

trap基因表达在表皮葡萄球菌生物膜形成及附属基因调节（agr）系统活化中的作用

目的研究表皮葡萄球菌trap基因表达与生物膜形成的相关性及其对agr系统的活化作用。方法采用微量板半定量法检测表皮葡萄球菌生物膜表型,二维电泳和高灵敏度的基质辅助激光解吸附飞行时间质谱分析比较表皮葡萄球菌标准株蛋白质表达谱,实时定量逆转录PCR检测trap基因和RNAⅢ的转录水平,CLUSTALX对表皮葡萄球菌trap基因与金黄色葡萄球菌序列进行分析．Mega软件构建进化树。结果比较分析形成生物膜的表皮葡萄球菌ATCC35984株与不形成生物膜的ATCC12228株生长中期的蛋白质组,发现ATCC35984株的TRAP蛋白量明显高于ATCC12228株。转录水平的检测显示ATCC12228株和生物膜阳性临床SE1457、SE671株的trap基因均低转录,但RNAⅢ的转录水平未降低。SE1457株agr系统突变后,RNAⅢ的转录明显降低,trap基因的转录无变化．RNAⅢ的转录随细菌生长变化而增加,trap基因转录保持不变．trap序列分析显示其在表皮葡萄球菌与金黄色葡萄球菌中较为保守,但存在明显的进化距离。结论在本论文所研究的表皮葡萄球菌菌株中,trap基因的转录和翻译与细菌生物膜的形成不存在直接的调控相关性;并提示TRAP蛋白不是表皮葡萄球菌agr系统活化所必需的细菌产物;表皮葡萄球菌trap基因序列与金黄色葡萄球菌之问存在差异,可能与其功能差异相关。     

表皮葡萄球菌临床菌株icaA、icaR基因与生物被膜形成关系的研究

目的研究表皮葡萄球菌icaA和icaR基因与生物被膜形成的相关性。方法收集湘雅三医院机械通气患者分离表皮葡萄球菌26株,按生物被膜成膜能力分成生物被膜阳性菌组和生物被膜阴性菌组各13株。采用RT-PCR分别对2组细菌icaA和icaR基因灰度比进行检测;采用real-time PCR分别检测2组细菌icaA和icaR基因的表达,并通过2-△△Ct方法进行相对定量。结果 RT-PCR结果显示icaA基因在生物被膜阳性组中的灰度比为0.96~1.38,在生物被膜阴性组中为0.89~1.56,表达差异无统计学意义(T=10.65,P=0.057);同理,icaR基因在生物被膜阳性组中的灰度比为0.79~1.24,在生物被膜阴性组中则为0.80~1.36,表达差异亦无统计学意义(T=14.62,P=0.479)。但icaA基因在生物被膜阳性组中的平均秩(16.35)明显高于生物被膜阴性菌组(10.65),而icaR基因生物被膜阳性组的平均秩(12.38)明显小于生物被膜阴性菌组(14.62)。real-time PCR检测发现表皮葡萄球菌生物被膜阳性组icaA基因的表达量(△CT=4.86)是生物被膜阴性组(△CT=13.57)的418.8倍(t=61.890,P0.01),而icaR基因的表达量(△CT=19.03)仅为生物被膜阴性组(△CT=11.85)的1/145(t=21.330,P0.01)。结论 icaA基因与表皮葡萄球菌生物被膜的形成呈正相关,而icaR基因与表皮葡萄球菌生物被膜的形成呈负相关。     

临床分离表皮葡萄球菌的大环内酯耐药性与生物膜形成能力、icaA基因型的关系

目的 研究68株临床分离表皮葡萄球菌大环内酯耐药性与生物膜形成能力、icaA基因型之间的关系,初步预测采用大环内酯抗菌药物预防表皮葡萄球菌生物膜感染的有效性.方法 以琼脂平板稀释法测定菌株的最小抑菌浓度,采用微孔测定法考察临床分离表皮衙萄球菌生物膜形成能力,通过PCR法测定icaA基因型,探讨三者之间的关系.结果 临床分离表皮葡萄球菌对大环内酯耐药程度较高(88.2%),且耐药菌生物膜形成能力较敏感菌显著增强(P<0.05),耐药菌与敏感菌的icaA阳性比率没有统计学差异(P>0.05).结论 红霉素、阿奇霉素、克拉霉素等常用的大环内酯类药物可能难以有效地预防表皮葡萄球菌生物膜感染.     

金黄色葡萄球菌临床分离株生物膜形成能力及其相关基因的检测

目的： 检测临床分离金黄色葡萄球菌（金葡菌）的生物膜形成能力及其相关基因。方法： 用半定量微量平板法检测临床分离的92株金葡菌生物膜形成能力,并通过扫描电镜观察生物膜形态,用PCR法来检测生物膜形成的相关基因ica和sarA。结果： 92株临床分离金葡菌菌株均可形成生物膜,PCR结果显示100%的分离株均含有sarA基因,78株（84.78%）含有ica基因。结论： 临床分离的金葡菌都具有形成生物膜的能力,生物膜形成的相关基因ica和sarA的携带率较高。     

耐甲氧西林凝固酶阴性葡萄球菌mecA基因检测分析

目的:调查耐甲氧西林凝固酶阴性葡萄球菌(MRCNS)耐药现状,分析其耐药表型与基因型的关系,为临床合理选用抗菌药物提供依据。方法:收集临床分离的86株凝固酶阴性葡萄球菌,采用头孢西丁纸片法检测耐药表型、聚合酶链反应(PCR)检测mecA基因,并将两种结果进行对比分析。结果:86株凝固酶阴性葡萄球菌中头孢西丁纸片法阳性64株,检出率为74.4%;mecA基因阳性58株,阳性率为67.4%;其中有1株头孢西丁纸片法阴性而mecA基因阳性,7株头孢西丁纸片法阳性而mecA基因阴性。结论:本地区耐甲氧西林凝固酶阴性葡萄球菌耐药状况严重;头孢西丁纸片法与PCR检测mecA基因一致率90.7%;PCR检测mecA基因可快速准确判定耐甲氧西林凝固酶阴性葡萄球菌。     

临床分离金黄色葡萄球菌生物膜相关基因的PCR分析

目的 确定临床分离的金黄色葡萄球菌生物膜形成能力和生物膜相关基因之间的关系,为生物膜形成的机制研究提供依据.方法 96孔板番红染色法检测生物膜的形成,icaAD、icaBC、sar、agr和sigB的PCR扩增,并对产物进行测序和同源性比较.结果 27株金黄色葡萄球菌中共有17株形成了肉眼可见的生物膜,其中以X387和X409两株最明显;有22株扩增出icaAD和icaBC,全部菌株扩增出sar、agr和sigB.结论 ica是形成金葡菌生物膜的关键基因,但ica在形成生物膜过程中需要其他基因的协同作用.     

呼吸道白念珠菌分离株生物膜表型差异的机制

摘要：目的探讨呼吸道白念珠菌分离株生物膜形成能力存在差异的可能分子机制。方法呼吸道白念珠菌临床分离株及标准株
ATCC90028体外粘附生长24 h形成生物膜,用XTT减低法测定其增殖情况,并分别提取生物膜态白念珠菌总RNA,用荧光定
量RT-PCR的方法测定转录因子CPH1、EFG1和粘附分子ALS3、HWP1基因的表达,采用△Ct的方法计算其相对表达量。结果
根据粘附生长的白念珠菌增殖情况,有8株临床株形成生物膜能力“高”,另7株临床株及标准株ATCC90028形成生物膜能力
“低”。生物膜表型差异的菌株间转录因子CPH1、EFG1的表达无显著差异（P>0.05）,而粘附分子ALS3、HWP1基因的表达有显
著差异（P<0.05）。结论呼吸道白念珠菌分离株生物膜形成存在表型差异,其差异的机制可能与转录因子CPH1和EFG1以外的
基因调控相关。
     

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.