非食源性腹泻中产气荚膜梭菌的毒力型分布特征

目的 探讨非食源性腹泻患者中分离的产气荚膜梭菌毒力型分布特征。方法 选取2020年1月1日至12月31日清华大学附属北京清华长庚医院收治的非食源性腹泻住院患者粪便样本分离的86株产气荚膜梭菌,其中59株分离于抗生素相关性腹泻（antibiotic-associated diarrhea, AAD）患者,27株分离于非AAD患者。采用实时荧光聚合酶链反应（polymerase chain reaction, PCR）检测菌株的6种毒素基因,即产气荚膜梭菌alpha毒素（cpa）、beta毒素（cpb）、epsilon毒素（etx）、iota毒素（ι-iap）、肠毒素（cpe）和坏死性肠炎B毒素（netB）,依据毒素类型将菌株分为A～G 7种毒力型,分析毒力型分布以及AAD组和非AAD组间毒力型的差异。结果 86株产气荚膜梭菌中,毒素基因cpa、cpb、etx、ι-iap、cpe和netB的检出率分别为100.0%、5.8%、0.0%、0.0%、18.6%和2.3%;A～G毒力型分别占74.4%、0.0%、5.8%、0.0%、0.0%、17.4%和2.3%。A毒力型在AAD组和非AAD组的...     

ICU病房的非艰难梭菌相关性医院感染腹泻

目的 了解三甲医院ICU病房非艰难梭菌医院感染腹泻患者产酸克雷伯菌和产气荚膜梭菌的感染率及临床特点,以期对ICU病房细菌性医院感染腹泻患者的诊治思路提供基础数据.方法 收集四川大学华西医院综合ICU病房2012年4～11月非艰难梭菌医院感染腹泻102例患者的粪便,PCR检测产酸克雷伯菌毒素基因pehX,产气荚膜梭菌菌种鉴定16S rRNA基因和毒素基因cpa、cpe,空肠弯曲菌菌种鉴定16S rRNA基因、mapA基因及毒素基因hi pO.PCR阳性产物双向测序并用工具BLAST算法进行比对.总结产毒菌株检测阳性的医院感染腹泻患者的临床特点.结果 102例非艰难梭菌医院感染腹泻患者的粪便中检出产毒的产酸克雷伯菌和产毒的产气荚膜梭菌各4例,各占非艰难梭菌相关性医院感染腹泻的3.9％.没有检出产毒的空肠弯曲菌.大多数检测阳性的患者都有严重的基础疾病,都在使用强有力干扰肠道正常菌群的抗菌药物,预后都很差.结论 尽管我院ICU病房非艰难梭菌相关医院感染腹泻的患者粪便中产毒产酸克雷伯菌和产气荚膜梭菌的检出率较低,但这两种菌感染的患者都有严重的基础疾病且预后不良.     

C型产气荚膜梭菌BL21(DE3)pXETAB2B1三价灭活苗的免疫效力试验

目的 研究C型产气荚膜梭菌BL21(DE3)(pXETAB281)三价灭活苗的免疫效力.方法 制备C型产气荚膜梭菌B121(DE3)(pXETAB281)三价灭活苗,免疫接种奶牛及ICR小鼠,分别进行静脉采血,观察抗血清中和C型产气荚膜梭菌强毒株C59-2毒素粗提物的情况.结果 奶牛抗血清中和C型产气荚膜梭菌强毒株C59-2毒素粗提物达到60%;ICR小鼠抗血清中和C型产气荚膜梭菌强毒株C59-2毒素粗提物达到100%.结论 C型产气荚膜梭菌BL21(DE3)(pXETAB281)三价灭活苗能中和C型产气荚膜梭菌强毒株C59-2毒素粗提物.     

产气荚膜梭菌β毒素分子生物学研究进展

产气荚膜梭菌 (Clostridiumperfringens )又名魏氏梭菌(cl welchii) ,是引起人和家畜肠道疾病的重要病因 ,广泛分布于土壤、污水、饲料、食物、人畜粪便及肠道中 ;该菌群属典型的条件致病菌 ,可以引起地区性流行 ,也有散发性报道 ,每年都有大量的动物因不同的产气荚膜梭菌感染而死亡。因此关于产气荚膜梭菌的分子生物学研究对疾病预防和畜牧生产具有重要的意义。根据其所分泌的肠毒素 (α、β1、β2 、ε、ι、enterotoxin)性质不同而分为A～E五型 ,其中对人致病的主要是A、C两型 ,B、C、D三型与动物的感染密切相关[1] 。C型是引起动物 …     

含产气荚膜梭菌肠毒素基因重组质粒pET-28a-CPE的构建和表达

目的 构建含全长产气荚膜梭菌肠毒素(CPE)基因的重组表达质粒pET-28a-CPE并进行原核表达.方法 培养并提取表达肠毒素的产气荚膜梭菌标准菌株64615的基因组DNA,PCR扩增出全长CPE基因片段并连接入pET-28a载体质粒中,构建重组原核表达质粒pET-28a-CPE,进行酶切及测序鉴定,并转化入感受态大肠杆菌(E.coli)BL21中,用IPTG诱导CPE表达.结果 复苏、培养成功产肠毒素的产气荚膜梭菌标准菌株64615,成功构建了表达质粒pET-28a-CPE,经酶切及测序鉴定结果 与设计的完全相符,成功用E.coil BL21进行了原核表达,目的 蛋白CPE占总表达蛋白45.37%.结论 本实验成功构建并表达了含CPE的重组质粒pET-28a-CPE,为进一步观察CPE时前列腺肿瘤等的生物学作用奠定了基础.     

一起由金黄色葡萄球菌及其肠毒素引发食物中毒的实验室检测与分析

目的查明引发食物中毒的致病菌,运用基因分型技术对致病菌进行溯源分析,为食物中毒诊断提供依据。方法采用食品安全国家GB 4789系列标准方法对致病菌进行分离、鉴定。金黄色葡萄球菌肠毒素分型检测采用酶联免疫吸附法(ELISA),肠毒素基因的检测采用实时荧光定量PCR的方法,利用脉冲场凝胶电泳(Pulsed-field gel electrophoresis,PFGE)进行同源性分析,全自动药敏接种判读仪(AIM-Vizion)进行耐药分析。结果 4份标本中都检测出金黄色葡萄球菌,4株菌均产A型肠毒素,携带sea基因,剩余食物分离株与病人分离株PFGE带型相似性为93.8%,具有高度同源性,菌株对5种抗生素耐药。结论本次食物中毒由肠毒素A型的金黄色葡萄球菌所致,实时荧光定量PCR可以弥补传统ELISA的不足。PFGE可迅速锁定传染源。提醒相关部门要加大合作和监督力度,保障食品安全。     

一起金黄色葡萄球菌引起食物中毒的病原溯源

目的分离鉴定病原菌,查明食物中毒原因;运用基因分型技术对分离菌进行溯源性分析。方法采用传统细菌学检测方法分离、鉴定病原菌,酶联免疫吸附法(ELISA)检测肠毒素分型,脉冲场凝胶电泳(PFGE)进行基因分型,用Bio Numerics软件进行聚类分析和病原溯源。结果此次食物中毒共分离到4株金黄色葡萄球菌,从患者肛拭样中分离出3株,从餐厅从业人员肛拭样中分离出1株,均携带同种金黄色葡萄球菌肠毒素(staphylococcal enterotoxin,SE),PFGE分型为同一带型,属同一克隆。结论此次食物中毒是由产SEA型肠毒素的同一克隆株金黄色葡萄球菌引起。     

院内感染艰难梭菌的毒素检测及核糖体分型研究

目的 了解艰难梭菌临床分离株的毒力情况,并运用核糖体分型技术进行流行病学研究.方法 采集住院腹泻患者的未成形粪便标本,无水乙醇处理后接种CDMN选择培养基行艰难梭菌分离培养,并通过革兰染色、耐氧试验和凝集试验行菌落鉴定;提取菌株DNA,选择特异性引物用PCR法扩增毒素基因tcdA和tcdB;核糖体分型针对细菌基因组16S～23S rDNA间区序列进行扩增,并根据电泳带型的多态性实现分型.结果 共计分离得44株艰难梭菌,3种毒素型分别为A+B+型、A-B+型和A-B-型,分别占 57％ 、34％和9％;18种核糖体型中以R8 型和R4 型为主,分别占 20％和18％.结论 本研究临床分离的艰难梭菌菌株中,毒素型以A+B+型为主,核糖体分型存在相对优势的型别;无证据提示存在院内艰难梭菌感染的暴发流行.     

破伤风患者的临床护理方法

特异性感染指由一些特殊的病菌、真菌等引起的感染,如结核杆菌、破伤风梭菌、产气荚膜梭菌。破伤风指破伤风梭菌侵入人体伤口后,生长繁殖,产生毒素所引起的一种急性特异性感染。     

产气荚膜梭菌α毒素亚单位基因的表达及表达产物活性分析

含有产气荚膜梭菌α毒素基因序列的克隆载体pMD18TCα ,经BamHⅠ和HindⅢ内切酶双酶切后 ,将α毒素亚单位基因亚克隆到原核表达载体pET30b中 ,构建表达载体pET30bPα。经酶切和测序鉴定 ,α毒素基因中的90 0bp的片段正确插入到表达载体中。重组菌株BL2 1(DE3)pET30bPα经IPTG诱导的表达产物经SDS PAGE和Western_blot分析 ,表达出大小为 38× 10 3的重组蛋白具有和α毒素阳性血清发生特异性的免疫反应。     

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.