革兰阴性杆菌β-内酰胺酶表型和基因型研究

目的了解铜陵地区临床分离的革兰阴性杆菌产生超广谱β-内酰胺酶(ESBLs)、头孢菌素酶(AmpC酶)和金属β-内酰胺酶(MBL)的分布情况,并检测产ESBLs大肠埃希菌和肺炎克雷伯菌的ESBLs基因型鉴定。方法采用三维试验检测革兰阴性杆菌产ESBLs和AmpC酶,纸片协同法检测MBL,用聚合酶链反应(PCR)对ESBLs表型阳性的大肠埃希菌和肺炎克雷伯菌进行TEM型、SHV型、CTX-M型三种耐药基因检测并克隆测序。结果356株革兰阴性杆菌检出产酶株90株,检出率25.3%。单产ESBLs革兰阴性杆菌57株,检出率16.0%,以肺炎克雷伯(31.6%)、大肠埃希菌(31.0%)检出率高;单产AmpC酶4株,检出率1.1%,其中阴沟肠杆菌3株;同时产ESBLs和AmpC酶16株,检出率4.5%,以鲍曼不动杆菌(12.5%)检出率最高;产MBL细菌13株,均为非发酵菌,以嗜麦芽窄食单胞菌检出率最高,为71.4%。17株ESBLs表型阳性的大肠埃希菌和肺炎克雷伯菌有16株检出ESBLs耐药基因,其中CTX-M型15株(88.2%),TME型13株(76.1%),SHV型2株(11.8%),有11株细菌同时带有两种ESBLs基因,1株带有3种ESBLs基因。结论革兰阴性杆菌产生ESBLs、AmpC酶和MBL多种β内酰胺酶,铜陵地区大肠埃希菌和肺炎克雷伯菌ESBLs基因型以CTX-M型为主。     

临床分离株中革兰阴性杆菌产超广谱β-内酰胺酶菌的分布

目的 了解临床分离的革兰阴性杆菌中产超广谱β-内酰胺酶(ESBLs)的分布及耐药性. 方法 对临床分离的革兰阴性杆菌,采用CLSI标准检测产ESBLs菌株. 结果 120株革兰阴性杆菌分离出38株产ESBLs菌,阳性率32%,包括大肠埃希菌、克雷伯菌属、阴沟肠杆菌、弗劳地枸椽酸杆菌、铜绿假单胞菌、鲍曼不动杆菌.其中,第一位是大肠埃希菌 (53.8%, 21 /38),其次产酸克雷伯菌(35.3,6/17)、肺炎克雷伯菌(33.3%,7/20);药敏结果显示除对碳青酶烯类抗生素敏感外,ESBLs阳性株对多种抗生素的耐药率均较高,且明显高于ESBLs阴性株. 结论 本院产ESBLs菌分离率较高,主要以大肠埃希菌、产酸克雷伯菌、肺炎克雷伯菌为主.     

革兰阴性杆菌TEM型β-内酰胺酶耐药基因研究

目的 了解中南大学3所附属医院产超广谱β-内酰胺酶(ESBLs)革兰阴性杆菌TEM型耐药基因的检出及流行情况.方法 收集254株多重耐药革兰阴性杆菌,采用双纸片协同法(DDS)进行ESBLs表型确证,多重PCR.法扩增blaTEM、blaSHV、blaOXA-1基因,TEM特异性引物对其整个开放阅读框进行扩增并对PCIL产物测序,通过BLAST分析确定基因型;并对携带有TEM基因的革兰阴性菌株用随机扩增多态DNA(RAPD)方法进行菌株的同源性分析.结果 经多重PCR扩增,105株肠杆菌获得阳性结果,其中85株菌携带TEM基因,26株菌携带SHV基因,10株菌携带OXA-1基因;6株非发酵菌获得阳性结果,均携带TEM基因,且全部为TEM-1型广谱β-内酰胺酶.携带blaTEM耐药基因的菌株所含RAPD类型:大肠埃希菌12种,肺炎克雷伯菌6种,阴沟肠杆菌7种.结论 TEM-1型β-内酰胺酶是湖南地区主要流行的TEM型酶;大肠埃希菌产TEM酶情况尤为突出;在同一医院以及同一病区存在TEM-1型β-内酰胺酶的克隆传播.     

泌尿系感染产超广谱β内酰胺酶(ESBLs)大肠埃希菌的耐药表型和基因分型

目的:了解本地区泌尿系感染产超广谱β内酰胺酶(ESBLs)大肠埃希菌耐药表型及其基因型分布.方法:收集2003-03/2004-06间临床分离的尿培养阳性大肠埃希菌共258株,ESBLs的检测采用NCCLs规定的ESBLs型筛选和确定实验,用K-B琼脂扩散法进行药物敏感性检测,并应用聚合酶链反应(PCR)方法对所分离的产ESBLs菌株进行SHV,CTX-M基因型检测.结果:产ESBLs大肠埃希菌检出率为24.0%(62株),67.7%(42株)携带CTX-M基因,14.5%(9株)的菌株携带SHV型基因,并有1.6%(1株)同时携带CTX-M基因和SHV型基因.结论:本地区泌尿系感染产ESBLs大肠埃希菌以耐头孢曲松和头孢噻肟为主;CTX-M型的ESBLs最为常见.     

血液中大肠埃希菌超广谱β-内酰胺酶耐药基因型研究

目的 了解本地区血液标本中产超广谱β-内酰胺酶(ESBLs)大肠埃希菌的分离率、耐药性和基因型.方法 收集临床血液标本分离的产ESBLs的大肠埃希菌26株,应用PCR基因扩增技术分别检测SHV、TEM、CTX-M、PER、CTX-M-2、Ampc、GES基因,DNA测序确定CTX-M、TEM基因型.结果 在26株菌株中15株为CTX-M型,占57.69%,11株为TEM型,占42.31%.结论 本地区临床血液标本分离的产ESBLs大肠埃希菌流行的主要基因型是CTX-M型和TEM型.     

消毒剂-磺胺耐药基因的检测

目的检测临床分离的革兰阴性杆菌菌株Ⅰ类整合子消毒剂-磺胺耐药(qacE△1-sul1)基因的携带情况。方法采用聚合酶链反应技术,对311株革兰阴性杆菌临床分离菌株(分别为大肠埃希菌96株,肺炎克雷伯菌肺炎亚种71株,铜绿假单胞菌64株,鲍氏不动杆菌61株,阴沟肠杆菌19株)进行qacE△1-sul1基因的检测。结果 311株革兰阴性杆菌中180株qacE△1-sul1基因阳性,大肠埃希菌、肺炎克雷伯菌肺炎亚种、铜绿假单胞菌、鲍氏不动杆菌、阴沟肠杆菌qacE△1-sul1基因阳性率分别为69.8%、53.5%、37.5%、75.4%、26.3%。大肠埃希菌和鲍氏不动杆菌qacE△1-sul1基因阳性率高于肺炎克雷伯菌肺炎亚种、铜绿假单胞菌和阴沟肠杆菌,差异有显著意义(q=3.019～6.129,P<0.05)。结论临床分离菌株qacE△1-sul1基因高携带率可能是目前医院感染的高发因素之一,临床应重视消毒剂的合理使用,并定期了解医院细菌耐消毒剂的情况。     

产ESBLs分离株耐药基因初步分型

目的了解产超广谱β-内酰胺酶（ESBLs）大肠埃希菌和克雷伯菌的主要基因型分布特点。方法用表型确证试验筛选出临床标本中产ESBLs的大肠埃希菌和克雷伯菌114株。应用PCR方法扩增产酶株的TEM、SHV和CTX-M基因。结果PCR结果显示TEM、SHV和CTX-M基因的总阳性率分别为40.4%、20.7%和83.3%,其中大肠埃希菌分别为48.1%、1.3%和93.5%,肺炎克雷伯菌分别为24.3%、81.1%和62.2%。46.8%的大肠埃希菌和51.4%的肺炎克雷伯菌同时携带多个基因。结论产ESBLs大肠埃希菌的主要基因型为CTX-M,肺炎克雷伯菌主要基因型为SHV。大多数菌株同时携带多个基因。     

大肠埃希菌超广谱β-内酰胺酶检测及耐药基因型分析

目的了解宁夏医科大学附属医院大肠埃希菌超广谱β-内酰胺酶（ESBLs）的分离率、耐药性和基因型。方法收集2008年1月-2008年12月住院患者分离的标本281株,采用K-B法进行药敏试验,采用PCR技术分别检测TEM、CTX-M基因。结果在281株大肠埃希菌中,有186株大肠埃希菌产超广谱ESBLs,产酶率为66.2%。在186株ESBLs菌株中有111株为TEM型,占59.7%,有66株为CTX-M型,占35.5%。结论产ESBLs株大肠埃希菌对常用抗菌药物的耐药性较为严重,而且存在多重耐药;本地区临床标本分离的产ESBLs大肠埃希菌流行的主要基因型是CTX-M型和TEM型。     

常见革兰阴性杆菌AmpC酶的检测分析

目的了解常见革兰阴性杆菌、阴沟肠杆菌、铜绿假单胞AmpCs发生率及表型构成,以及产ESBLs的大肠埃希菌、肺炎克雷伯菌AmpC酶的发生率及表型构成。方法采用底物协同-拮抗法(MSSAT)检测大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌、鲍氏不动杆菌、阴沟肠杆菌产AmpCs情况及表型构成。结果产ESBLs的大肠埃希菌和产ESBLs的肺炎克雷伯菌、铜绿假单胞菌、鲍氏不动杆菌、阴沟肠杆菌AmpC酶检出率分别40%、56%、100%、100%、100%。结论产ESBLs大肠埃希菌和肺炎克雷伯菌同时存在AmpC酶的机制,应引起临床和实验室工作者的重视。AmpC酶的耐药表型与菌株的耐药程度相关。     

2O3株产超广谱β-内酰胺酶菌的检测及耐药性

目的:了解革兰氏阴性杆菌产超广谱β-内酰胺酶(ESBLs)的情况及耐药性,以利于对产ESBLs菌株的治疗.方法:采用NCCLS推荐的确认法.结果:727株革兰阴杆性菌中,共检出产ESBLs菌203株,检出率为27.9%,其中大肠埃希菌87株、克雷伯杆菌72株、嗜麦芽窄食单胞菌17株、费劳地枸橼酸菌11株、阴沟肠杆菌5株、鼠伤寒沙门菌4株、产气肠杆菌2株、铜绿假单胞菌2株、亲水气单胞菌2株、聚团肠杆菌1株;亚胺培南除对1株产ESBLs大肠埃希菌和17株产ESBLs嗜麦芽窄食菌耐药外,对其他产ESBLs菌均表现出最强的抗菌作用;头孢哌酮/舒巴坦对产ESBLs嗜麦芽窄食菌、大肠埃希菌及费劳地拘橼酸菌有较好的抗菌作用;丁胺卡那霉素对产ESBLs大肠埃希菌呈现较好的抗菌作用.结论:我院革兰阴性菌产ESBLs情况严重,而且已经分离出对亚胺培南耐药的ESBLs大肠埃希菌,临床医生应尽量在确立了病原学诊断后参考细菌药敏报告用药,以减少耐药菌株的产生.     

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.