稳定表达荧光素酶的产酸克雷伯菌在评估不同灭菌方法中的应用

目的构建稳定表达荧光素酶基因（lux）的产酸克雷伯菌,旨在更直观地观察产酸克雷伯菌（K. oxytoca）在宿主体内的分布以及评估不同杀菌方法的效果。 方法扩增pBAV1k-T5-Lux质粒的荧光素酶基因簇（Lux A/B/C/D/E）,构建含有Lux A/B/C/D/E基因簇的pBBR1质粒（pBBR1-lux）,并获得能够表达荧光素基因基团的大肠埃希菌（E. coli-pBBR1-lux）。将pBBR1-lux质粒电转化至产酸克雷伯菌感受态细胞,经荧光强度鉴定及菌株的3次传代,筛选能够稳定表达lux基因的产酸克雷伯菌（K. oxytoca-pBBR1-lux）。 结果连接成功的环状质粒产物命名为pBBR1-lux。与大肠埃希菌对照株相比,含E. coli-pBBR1-lux的Luminesence荧光信号值显著升高[15 345（14 676,18 654） vs. 63（60,82）],差异具有统计学意义（t = 21.14、P = 0.035）。K. oxytoca-pBBR1-lux的Luminesence荧光信号值[399 303（265 245,617 192）]显著高于产酸克雷伯菌对照菌株[83（63.5,86.75）],差异具有统计学意义（t = 7.07、P = 0.014）。E. coli-pBBR1-lux和K. oxytoca-pBBR1-lux均能够在Veritas微孔板光度计和小动物成像仪上检测到Luminesence荧光信号。将所得菌株按照1︰10稀释6个梯度,荧光值检测结果显示,Luminesence值随菌落浓度降低而下降。多次传代后pBBR1-lux能够在产酸克雷伯菌中稳定表达荧光素酶,不同理化杀菌方法对产酸克雷伯菌杀菌效果不同,紫外线和84消毒液（10%）是最有效的杀灭产酸克雷伯菌方法。 结论本研究获得了可被微孔板光度计和小动物成像仪检测到的稳定表达荧光素酶的K. oxytoca-pBBR1-lux。     

产ESBLs肺炎克雷伯菌引起前列腺增生患者膀胱炎1例

产超广谱β-内酰胺酶(ESBLs)的肺炎克雷伯菌引起临床上的各种感染常见[1],但引起男性膀胱炎的较少见.2001年3月～2002年6月,从我院1例膀胱炎反复发作的前列腺增生患者中段尿、膀胱穿刺尿中多次分离出此菌.现报告如下.     

路邓葡萄球菌致新生儿败血症一例

患儿男,于2012年1月27日在中国人民解放军总医院行剖宫产娩出,羊水清,其母孕期平稳,无妊娠合并症.患儿出生后混合喂养,吃奶可,无明显呛吐奶.2月1日无明显诱因出现咳嗽,喉中痰响,体温正常.2月2日就诊于中国人民解放军总医院门诊,考虑"上呼吸道感染",予猴枣散(0.15g/次,2次/d),乙酰半胱氨酸颗粒(0.1g/次,2次/d),口服治疗.2月3日仍有咳嗽,出现吐沫,体温37.0～ 37.8℃,纳差较前差,粪便略稀,5～6次/d.     

肺炎克雷伯菌肝脓肿临床特征分析

细菌性肝脓肿是肝脏的严重感染性疾病,其发病机制尚不十分清楚,临床表现不典型,易导致误诊和漏诊.多项研究证实,肺炎克雷伯菌已成为目前细菌性肝脓肿的主要致病菌[1-2].为加强对该病的认识,现回顾性分析本院65例细菌性肝脓肿患者的临床资料,比较肺炎克雷伯菌和非肺炎克雷伯菌肝脓肿在临床特征、伴随疾病及诊疗等方面的差异,为临床诊疗及判断预后提供一定的帮助.     

肺炎克雷伯菌与植生克雷伯菌喹诺酮类耐药基因研究

目的 分析宁波大学医学院附属医院克雷伯菌属的耐药性及其对喹喏酮类药物的耐药机制.方法 收集2009年10月-2011年3月分离出的20株克雷伯菌属菌株,经鉴定确认18株为肺炎克雷伯菌,2株为植生克雷伯菌.采用k-b纸片扩散法检测其对药物的敏感性.pcr法检测染色体介导的gyra、parc基因和质粒介导的aac(6’)-Ⅰ b-cr、qnra、qnrb、qnrs、qepa基因,pcr阳性产物采用pcr直接全自动荧光法测序.结果 20株克雷伯菌属菌对β-内酰胺类、氨基糖苷类、喹诺酮类抗菌药物均表现为多重耐药性(耐药率均在80％以上),其中植生克雷伯菌对亚胺培南和美罗培南敏感.18株(90％)菌株存在gyra、parc基因突变;aac(6’)-Ⅰ b-cr阳性12株(60％),qnrb和qnrs阳性均为4株(20％).结论 本组20株克雷伯菌属菌对喹诺酮类药物耐药主要与gyra和parc基因突变相关. abstract： objective to investigate the multi-drug resistance of klebsiella strains and its mechanism.methods twenty strains of klebsiella were isolated from the affiliated hospital of medical college,ningbo university from october 2009 to march 2011,in which 18 isolates were klebsiella pneumonia and 2 were klebsiella planticola. drug sensitivity was determined by k-b tests. drug resistant genes gyra,parc (chromosome mediated) and aac( 6′)-i b-cr,qnra,qnrb,qnrs,qepa (plasmid mediated) were amplified by pcr and verified by direct automated fluorogenic sequencing. results resistance to β-1actams,aminoglycosides and quinolones was observed in 20 strains,and resistant rates were all above 80％.klebsiella planticola strains were sensitive to imipenem and meropenem.mutations of gyra and parc genes existed in 18 strains (90％),and the positive rates of aac (6') -i b-c r,qnrb and qnrs were 60％ (12/20),20％ (4/20) and 20％ (4/20),respectively.conclusion the mutations ofgyra and parc genes may be the main cause of the resistance to quinolones in these strains.     

侵袭性肺炎克雷伯菌肝脓肿综合征临床及影像学特征

目的 观察侵袭性肺炎克雷伯菌肝脓肿综合征（IKPLAS）临床及影像学特征。方法 回顾性纳入68例肺炎克雷伯菌肝脓肿（KPLA）患者,根据是否发生肝外侵袭性感染分为IKPLAS组（n=25）及非IKPLAS组（n=43）,比较组间临床资料及CT和/或MRI表现。结果 IKPLAS组患者年龄低于而糖化血红蛋白、D-二聚体均高于非IKPLAS组（P均＜0.05）。IKPLAS组18例、非IKPLAS组6例可见肝静脉血栓性静脉炎,IKPLAS组10例、非IKPLAS组28例可见动脉期脓肿周围异常高灌注,组间差异均有统计学意义（P均＜0.05）。IKPLAS组肝外感染主要累及肺（19/25,76.00%）。结论 IKPLAS患者年龄低于而糖化血红蛋白、D-二聚体高于其他KPLA;较常见肝静脉血栓性静脉炎及肝外感染主要累及肺为IKPLAS影像学特征。     

新生儿细菌性痢疾的临床特点

细菌性痢疾是痢疾杆菌引起的肠道传染病,多流行于夏秋季节,是儿科的常见病,严重影响患儿的健康。细菌性痢疾在新生儿期发病罕见,国内鲜有报道,临床特点尚不十分清楚。本研究对2006年1月至2009年12月安徽省儿童医院收治的新生儿细菌性痢疾住院患儿的临床资料进行分析,探讨新生儿细菌性痢疾的临床特点。     

产β-内酰胺酶肺炎克雷伯菌新型整合酶基因研究

老年患者肺炎克雷伯菌β-内酰胺酶基因和氨基糖苷类药物耐药相关基因已有较多报道[1-2].为进一步了解老年人肺炎克雷伯菌分离株转座子和整合子的存在状况,笔者对20株肺炎克雷伯菌进行了转座子遗传标记（tnp513、tnpU 、tnpA、merA）和整合子遗传标记（int Ⅰ1、int Ⅰ2、intⅠ3）检测.     

产多种β-内酰胺酶肺炎克雷伯菌的基因型研究

目的 研究医院感染肺炎克雷伯菌的β-内酰胺酶基因型.方法 连续收集从临床住院患者中分离的肺炎克雷伯菌,采用接合试验、PCR、PCR产物测序、等电聚焦试验、超广谱β-内酰胺酶（ESBLs）确认试验等方法明确基因型.结果 共收集75株肺炎克雷伯菌,48株（64.0%,48/75）含β-内酰胺酶基因,其中ESBLs占52.0%（39/75）.在48株产酶菌株中,携带2种基因型的占35.4%（17/48）,携带3种基因型的占14.6%（7/48）,携带4种基因型的占10.4%（5/48）.CTX-M型β-内酰胺酶基因的检出率最高（30/48,62.5%）,其次为TEM型（26/48,54.2%）和SHV型（25/48,52.1%）.产DHA-1型AmpC酶的肺炎克雷伯菌有9株,且有8株与ESBLs并存.同时还检测到3株产KPC-2型碳青霉烯酶肺炎克雷伯菌.ESBLs表型确认试验结果的假阴性率为23.1%（9/39）.结论 医院感染肺炎克雷伯菌的β-内酰胺基因分布复杂,临床上呈多药耐药状态.     

嗜麦芽窄食单胞菌败血症的临床及预后危险因素分析

近年来,嗜麦芽窄食单胞菌的分离率逐渐上升[1],由于该菌耐药机制复杂,嗜麦芽窄食单胞菌败血症的病死率高[2].本研究分析了26例嗜麦芽窄食单胞菌败血症患者的相关临床资料及药敏试验结果,以期为临床感染控制及合理用药提供依据.     

New ABCC8 mutations in relapsing neonatal diabetes and clinical features

Activating mutations in the ABCC8 gene that encodes the sulfonylurea receptor 1 (SUR1) regulatory subunit of the pancreatic islet ATP-sensitive K(+) channel (K(ATP) channel) cause both permanent and transient neonatal diabetes. Recently, we have described the novel mechanism where basal Mg-nucleotide-dependent stimulatory action of SUR1 on the Kir6.2 pore is increased. In our present study, we identified six new heterozygous ABCC8 mutations, mainly in patients presenting the transient form of neonatal diabetes (six of eight), with a median duration of initial insulin therapy of 17 months (range 0.5-38.0). Most of these mutations map to key functional domains of SUR1. Whereas Kir6.2 mutations are a common cause of permanent neonatal diabetes and in a few cases associate with the DEND (developmental delay, epilepsy, and neonatal diabetes) syndrome, SUR1 mutations are more frequent in transient (52%) compared with permanent (14%) neonatal diabetes cases screened for ABCC8 in our series. Although ketoacidosis is frequent at presentation, SUR1 mutations associate mainly with transient hyperglycemia, with possible recurrence later in life. One-half of the SUR1 neonatal diabetic patients presented with de novo mutations. In some familial cases, diabetes is not always present in the adult carriers of SUR1 mutations, supporting variability in their clinical expressivity that remains to be fully explained.     

包皮环切术的并发症及防范

目的：总结本组资料的并发症以利预防和正确施术。方法：分析本组的并发症,结合文献提出防范措施。结果：1200例环切术共发生并发症36例,发生率为3%。并发症主要有疼痛、出血和水肿,也包括感染和切口部分裂开,龟头表面点状灼伤以及早期形态不良。这些并发症中,出现3例严重并发症。其中2例出血,经再次手术痊愈;另一例严重水肿,经保守治疗无效,长达9个月,最后经＂纵切横缝＂治愈。结论：术者应熟知环切术的并发症,术前应告知患者或患者（小儿）双亲正确和公正的信息,共同讨论决定手术。＂纵切横缝＂是处理术后严重水肿病例简便而有效的措施。     

Cardiac aging phenomenon and its clinical features by echocardiography

Even in healthy subjects, the heart ages along with other organs of the body. A prominent change is progressive left ventricular (LV) diastolic dysfunction, even though LV mass increases slightly during aging. Accordingly, assessment of LV dysfunction can be employed as a surrogate marker of cardiac age. The clinical factors that may accelerate the cardiac aging process include visceral obesity, diabetes mellitus, dyslipidemia, and hypertension. At the molecular level in cardiac myocytes, reactive oxygen species, transforming growth factor-β, mitochondrial function, and lysosomal function are also related to cardiac age. Furthermore, age-related LV dysfunction has been shown to be one of the main risk factors for future heart failure. Consequently, assessment of LV diastolic function is necessary for both preventing cardiac events and assessing cardiac age. Echocardiography provides a noninvasive assessment of cardiac structure and function. This review describes how to assess cardiac aging using echocardiography, and how to interpret the clinical relevance of the findings.     

Hemodynamic features of hypotension induced by nicardipine in dogs

Hemodynamic features of hypotension induced by nicardipine were studied in 18 mongrel dogs under 0.87% halothane in oxygen (1MAC). They were randomly allocated to one of two groups. Group C received no vasodilator therapy and served as control and group N received infusion of nicardipine. Mean arterial pressure decreased and was maintained at 60 mmHg for 60 minutes in group N. No change was noted in hemodynamic variables measured in group C throughout the experiment. During and after induced hypotension in group N, cardiac index (CI), stroke volume index (SVI), central venous pressure (CVP), and right ventricular stroke work index (RVSWI) increased significantly compared with the control values. On the other hand, systemic vascular resistance (SVR) was significantly reduced, reaching 25% of the control value at the end of hypotension period. Heart rate showed a progressive increase but not significantly. Left ventricular maximum dp/dt showed a moderate increase during hypotension, but then decreased gradually to the control value after induced hypotension. Left ventricular stroke work index (LVSWI) and pulmonary vascular resistant (PVR) were unchanged from the control value during and after induced hypotension. The data indicate that nicardipine is a potent systemic vasodilator with hyperdynamic hemodynamic effects in addition to significant increase in right ventricular function.     

Pathogenesis and clinical features of acute cholangitis accompanied by shock

We investigated 91 cases of acute cholangitis, including 42 of severe cholangitis and 49 of mild cholangitis. The incidence of endotoxemia was 78.6 percent in 42 and 32.6 percent in the 49 patients. In the 42 with severe cholangitis, remarkable leukocytosis, thrombocytopenia, decrease of serum CH50, C3, plasma fibronectin and phagocytic index were characteristic. Disseminated intravascular coagulation (DIC) was observed in 76.2 percent. Since there was a positive correlation between platelet counts and levels of CH50 and C3, the decrease of platelet count, and the occurrence of DIC in patients with endotoxemia were thought to be closely related to the consumption of complements. There was no difference in mortality rate between nonsurgical treatment (57.8 percent) and the emergency bile drainage treatment (56.5 percent). The results of therapy depended on the degree of complicating DIC. We conclude that acute cholangitis was aggravated by endotoximia and that severe cholangitis was accompanied by DIC induced by a decline in phylaxis.