胆固醇结石、胆囊黏膜和胆汁细菌DNA的检测

目的　探讨胆固醇结石中细菌DNA的意义。方法　用半定量PCR和16SrRNA序列分析法检测胆囊黏膜、胆汁和胆石的细菌DNA ,观察围手术期抗生素的使用对检测结果的影响。结果　(1)结石核心、外周、胆汁和胆囊黏膜细菌DNA阳性率分别为79%、82 %、77%和6 4 %。胆石、胆汁和胆囊黏膜间细菌DNA阳性率无相关性。(2 )结石核心、外周、胆汁和胆囊黏膜细菌菌落数对数值(cfu)分别为3.19±2 .0 9、3.2 6±2 .0 5、3.2 3±2 .14和3.2 8±2 .70。结石内外菌落数相关系数为0 .82 2 (P <0 .0 5 )。胆汁和胆囊黏膜菌落数不相关。胆石核心及外周与胆汁和胆囊黏膜菌落数均不相关。(3)术前用与不用抗生素者胆汁、胆囊黏膜、胆石核心和外周的细菌DNA阳性率均差异无显著性。但用抗生素者胆汁菌落数下降,与未用抗生素组比较差异有显著性(P <0 .0 1) ,而胆囊黏膜、胆石核心和外周的菌落数则差异无显著性。(4)胆石、胆汁、胆囊黏膜的细菌DNA序列并不完全相同,胆石核心与外周部位的DNA序列也不相同。结论　胆固醇结石中细菌DNA可能并非胆道环境细菌污染的结果。较之胆汁和胆囊黏膜的细菌DNA ,胆固醇结石中的细菌DNA与结石形成的关系更密切。     

胆囊结石与胆囊壁黏膜幽门螺杆菌定植的关系

目的:研究和探讨幽门螺旋杆菌(Helicobacter pylori,H.pylori)在胆囊壁黏膜中的定植与胆囊结石形成的相关性.方法:分别对结石性胆囊炎和非结石性胆囊切除患者的胆囊组织和胆汁进行幽门螺杆菌检测,采用多聚酶链式反应检测幽门螺杆菌特异性DNA片段、酶联免疫吸附试验检测幽门螺杆菌抗体蛋白IgG、并对两组胆囊黏膜标本行幽门螺杆菌免疫组织化学技术染色检测.结果:采用ELISA方法检测结石组和非结石组中H.pylori抗体蛋白,结石组中48.44%阳性,非结石组43.75%阳性,两者之间无统计学差异.采用PCR方法检测结石组和非结石组胆囊黏膜和胆汁标本中H.pylori特异性DNA片段,胆囊黏膜中结石组25例阳性(39.06%),非结石组2例阳性(12.50%),两者之间有统计学差异(P=0.04);而胆汁中结石组59.38%阳性,非结石组56.25%阳性,两者之间无统计学差异.对两组胆囊黏膜标本行免疫组织化学检测,结石组有19例阳性,在胆囊黏膜腺腔中可见棒状、弯曲杆状或球形的菌体,呈棕黄色或棕褐色.而非结石组仅有1例检出.结论:胆道中H.pylori感染在胆囊结石形成的过程中不起主要作用,而H.pylori在胆囊黏膜的定植则参与胆囊结石的形成.     

老年病人胆道疾病的厌氧菌感染

对胆道感染的需氧菌已进行充分研究,最常见的为大肠杆菌、大肠菌类和肠球菌,但对厌氧菌注意不够。大多认为,产气荚膜杆菌是胆道感染最常见的厌氧菌,其他厌氧菌如固紫阳性厌氧球菌、乳酸杆菌和放线菌则较少见。作者应用改良的厌氧培养方法进行胆汁的细菌学研究。 52例作胆道手术病人,男19,女33,平均年龄73.4岁,其中胆囊炎7例,胆囊炎伴胆囊结石34例,胆道癌9例,胆总管结石2例。3例术前未用抗菌素。手术时直接胆囊穿刺抽取胆汁做细菌学检查。结果: 52例胆汁培养,12例无菌生长,40例有细菌生长,其中22例单纯为需氧菌感染,18例为需氧菌与厌氧菌混合感染。除8例系单一菌种感染,其余32例均为多种细菌感染。胆汁培养最常见细菌是大肠杆菌(30例),其次是克雷白氏-肠杆菌群(26例)。第三常见的细菌是脆弱厌氧杆菌(15例,占29％),它系最常见的厌氧     

胆囊结石患者胆汁、粘膜和结石需氧菌和厌氧菌脱氧核糖核酸研究

目的研究胆囊结石患者胆汁、粘膜和结石细菌脱氧核糖核酸(DNA),提供细菌在胆囊结石形成中的分子生物学汪据.方法采用聚合酶链式反应(PCR)对70例胆汁、粘膜和结石标本中5种需氧菌包括大肠埃氏杆菌、克雷伯杆菌、幽门螺杆菌、变形杆菌、伤寒杆菌和4种厌氧菌包括脆弱类杆菌、消化球菌、产气荚膜杆菌、梭形芽胞杆菌DNA进行检测.结果胆汁、粘膜、结石和门静脉血中需氧菌和厌氧菌DNA存在率分别为80%,86%,61%,36%和33%,50%,24%,46%;两类细菌DNA共同存在分别为29%,44%,16%,14%.结论胆汁、粘膜、结石中存在多细菌DNA,胆结石核心形成可能与细菌感染有关.     

肝螺杆菌感染与人类原发性肝细胞癌的相关性研究

目的本研究旨在了解肝肠源性螺杆菌是否存在于肝细胞癌患者的胆汁内,探讨此菌与肝癌的相关性。方法收集经病理确诊的47例原发性肝细胞癌患者及24例结肠直肠癌肝转移患者的胆汁标本。使用特定16SrRNA引物的聚合酶链式反应(PCR)及DNA测序检测胆汁标本中的螺杆菌属及肝螺杆菌特异性16SrRNA,并使用酶联免疫吸附法(ELISA)检测胆汁标本中的肝螺杆菌抗体。结果 47例原发性肝细胞癌患者胆汁标本中有12例(25.5%)检测出肝螺杆菌特异性16SrRNA,15例(31.9%)检测出抗肝螺杆菌抗体;24例结肠直肠癌肝转移患者胆汁标本中只有1例(4.2%)检测出肝螺杆菌特异性16SrRNA基因,3例(12.5%)检测出抗肝螺杆菌抗体(P<0.05)。结论原发性肝细胞癌患者可检测出肝螺杆菌,这可能与肝细胞癌的发病机制有一定联系。     

聚合酶链式反应和常规培养检测胆总管结石中细菌及其意义的研究

探讨细菌在胆总管结石形成中的作用。通过ERCP导管抽取胆汁进行需氧菌培养。同时用PCR方法对结石内大肠杆菌DAN进行检测。56例胆总管结石胆汁标本需氧菌培养中有53例检出细菌,检出率为94.64%,埃希氏菌属、克雷伯氏菌属、肠杆菌属、肠球菌属为胆道主要致病菌。11例胆总管结石中,3例大肠杆菌DNA扩增阳性。导管取材进行细菌培养结果可信;细菌感染在胆总管结石形成过程中起核心作用。     

聚合酶链反应检测胆囊黏膜、胆石、胆汁中幽门螺杆菌DNA结果分析

多年来一直认为幽门螺杆菌 (Ｈｅｌｉｃｏｂａｃｔｅｒｐｙｌｏｒｉ,Ｈｐ)只定植于胃 ,引起急慢性胃炎和消化性溃疡 ,并与胃癌有关。随着对Ｈｐ生物特性及其致病性的深入研究 ,已发现有几种螺杆菌可引起肝胆系统疾病[1] 。为了进一步研究Ｈｐ与胆石形成的关系 ,我们采用ＰＣＲ在胆囊黏膜、胆石、胆汁中检出ＨｐＤＮＡ物质 ,现将结果报告如下。一、资料和方法(一 )标本来源1999年 11月～ 2 0 0 0年 4月收集在我院外科因胆石症行胆道手术 5 0例的标本 ,男 15例 ,女 35例。平均年龄 5 1岁 (2 1～ 77岁 ) ,其中胆囊结石 36例 ,胆总管结石梗阻性…     

肝癌患者肝组织中螺杆菌感染的研究

目的　探讨螺杆菌感染是否与人类肝癌的发生相关。方法　选取经病理诊断的肝癌组织及癌旁组织 2 0例为研究对象 ,非肝癌组织 16例作对照 ,采用聚合酶链反应 (PCR)扩增螺杆菌 16SrRNA基因 (16SrDNA)检测螺杆菌 ,扩增产物经Southern杂交证实 ,并对部分标本的PCR产物进行测序及同源比较。阳性者再扩增幽门螺杆菌 (H pylori)的特异基因 (2 6 -kDa蛋白 )和相关功能基因 (cagA、vacA、glmM、rps4 )来验证是否为该菌。结果　 4 0 % (8/ 2 0 )的肝癌组织中发现螺杆菌 16SrD NA存在 ,其对应的癌旁组织大部分亦阳性 (7/ 8) ,而非肝癌组无一例阳性 (P <0 0 1)。所有PCR产物用Southern杂交得到证实。 6例测序及同源比较显示 ,肝癌组织中的螺杆菌 16SrDNA序列与H pylori的 16SrDNA序列有 98 5 %～ 99 0 %的同源性。阳性标本中 2 6 -kDa基因大部分亦阳性 (7/ 8) ,但仅 2例扩增出cagA基因 ,2例扩增出glmM基因 ,一直未扩增出vacA基因和rps4基因。 结论　肝癌患者肝组织中螺杆菌感染率较高 ,它与肝癌的发生可能存在某种联系。     

胃黏膜组织中恶臭假单胞菌临床意义的探讨

目的探索恶臭假单胞菌在幽门螺杆菌感染相关胃肠疾病中的意义。方法采集14 C尿素呼气试验阳性患者病变胃黏膜354例,进行细菌的分离培养。根据菌落形态、革兰染色、尿素酶试验及幽门螺杆菌特异性16SrRNA基因片段的PCR进行幽门螺杆菌的鉴定,同时提取胃黏膜组织DNA,通过幽门螺杆菌特异性PCR进行快速诊断。将非幽门螺杆菌转种于营养琼脂培养基,进行快速尿素酶试验。尿素酶阳性的非幽门螺杆菌染色体DNA利用细菌16SrRNA基因通用引物进行PCR扩增、测序及序列比对。使用全自动细菌鉴定仪对经测序比对为恶臭假单胞菌的菌株进行鉴定。采用K-B纸片扩散法进行药物敏感性试验。结果从354例样本中分离出革兰阴性、尿素酶阳性的恶臭假单胞菌10株,经16SrRNA基因测序和序列比对,与GenBank中恶臭假单胞菌的相似性≥98%。10株恶臭假单胞菌的胃黏膜标本中,6例标本幽门螺杆菌特异性PCR为阳性,4例为阴性。传代存活的7株恶臭假单胞菌的K-B法药物敏感试验显示对左氧氟沙星均敏感,1株对四环素敏感,5株对阿莫西林耐药,6株对克拉霉素耐药,7株对甲硝唑、氨苄青霉素均耐药。7株菌经全自动细菌生化鉴定仪鉴定结果均为恶臭假单胞菌。结论病变胃黏膜中分离出尿素酶阳性且对治疗幽门螺杆菌感染的多种抗生素耐药;可能造成临床上14 C-尿素呼气试验假阳性,并继发感染影响胃肠疾病的进展。     

褐色素结石的胆汁酸成分

褐色素结石不仅在其成分上而且在其病因学上均不同于胆固醇和黑色素结石;分析褐色素结石的成分可以提供成石机制的线索。作者用气-液相层析法分析褐色素结石中的胆汁酸含量,并将其与胆汁进行比较。研究的胆石标本全部是胆囊结石。28例为褐色素结石,10例为胆固醇结石,5例为黑色素结石。胆囊胆汁在探腹时由穿刺吸引获得,对照组的胆汁标本在胃癌或结肠癌患者手术时用细针吸出获得,以作需氧菌和厌氧菌培养。胆石干燥、研粉、秤重,并用二甲基硫氧-丙酮-盐酸的混合液提取。洗提液浓缩后用二乙胺羟丙基葡聚糖凝胶LH-20柱状层析,将胆汁酸分为游离、甘     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

Study of constancy of hydrogen-consuming flora of human colon

The constancy of the hydrogen consuming flora of the human colon was studied in 15 healthy subjects via two measurements obtained 18 to 36 months apart. Hydrogen disappearance rate and the major products of H₂-consuming bacteria, methane and sulfide, were measured during incubation of fecal homogenates with excess hydrogen and sulfate. In 11/15, the hydrogen consumption rate and the predominant hydrogen-consuming pathway (methanogenesis, sulfate reduction, or neither) remained constant. However, major shifts in these pathways were observed in four subjects, with two losing and two gaining the ability to produce methane. Methanogenesis was associated with the highest hydrogen consumption rate. This study demonstrates that clinically unrecognizable, major alterations of the colonic flora occur in healthy subjects. Understanding of the factors responsible for these alterations might allow for therapeutic manipulation of the colonic flora.Supported in part by the Department of Veterans Affairs and NIDDKD RO1 DK 13309-25.