套式聚合酶链反应扩增胆固醇结石中细菌DNA及其临床意 …

为探讨细菌在胆固醇结石病机理中的作用，从３０例胆汁细菌培养阴性的胆囊胆固醇结石中提取ＤＮＡ，应用套式聚合酶链反应扩增细菌１６ＳｒＲＮＡ基因片段作为有细菌存在的分子证据。结果显示，２６例胆固醇结石中有细菌ＥＮＡ存在。     

由胆固醇结石提取DNA进行套式聚合酶链反应的研究

为探讨微生物在胆固醇结石形成中的作用，作者从３０例胆汁细菌培养阴性的胆囊胆固醇结石中提取ＤＮＡ，应用套式聚合酶链反应（ＮＰ－ＰＣＲ）技术从中特异性地扩增细菌ＤＮＡ片段。结果显示２６例（８６．６７％）胆固醇结石中有细菌ＤＮＡ存在。此外，用１６ＳｒＲＮＡ基因序列对照分析鉴定细菌种类，８例（２６．６７％）为与大肠杆菌相似的ＤＮＡ片段；７例（２３．３３％）为痤疮丙酸杆菌型ＤＮＡ序列；２例（６．６７％）ＤＮＡ片段与化脓性链球菌相关；７例（２３．３３％）ＤＮＡ片段具有多样性，可能有多种细菌混合感染；另外２例（６．６７％）ＤＮＡ分子量较低，归类于其它未鉴定细菌。作者认为多数胆固醇结石内有细菌ＤＮＡ存在，但这些微生物的确切作用，有待于进一步深入研究。     

胆固醇结石中细菌的电镜观察

目的 通过电镜观察胆固醇结石内细菌的存在。方法 选取我院接受手术治疗的12例胆石症患者，男6例，女6例，平均年龄54．6岁(35∽72岁)，其中胆囊结石5例，肝内外胆管结石1例，胆总管结石并胆囊结石6例。利用透射电镜对结石进行细菌学观察。结果 在胆固醇结晶中和结石核心均发现有细菌样结构，呈卵圆形及长卵圆形，菌壁样结构不清晰，内部结构基本消失，细胞器、细胞核等结构均不可见。结论 胆固醇结石中有细菌样物质存在，位于结石的核心和外周，提示细菌在胆固醇结石的形成中可能充当了始动因子的作用。     

胆固醇结石中的细菌及其在成石机制中的作用

目的: 通过电镜观察胆固醇结石内的细菌形态学变化,探索细菌在成石中的作用.方法: 选择接受手术治疗的病人共计46例,男24例,女22例;平均年龄54.6岁(35~72岁);均确诊为胆石症.其中胆囊结石20例,肝内外胆管结石12例,胆总管结石并胆囊结石14例.利用扫描电镜、透射电镜对胆固醇结石进行观察.结果: 胆固醇结石表层结构内均能够发现细菌,胆固醇结晶中少有细菌,结石核心可发现细菌.结论: 对胆固醇结石进行扫描电镜、透射电镜观察,发现胆固醇结石中有细菌存在,细菌存在于结石的核心和外周,说明细菌在胆固醇结石的成因和发病机制中可能充当始动因子作用.     

细菌与胆石症的关系

１９６６年Ｍａｋｉ〔１〕提出了β葡萄糖醛酸酶学说,认为大肠杆菌可以分泌β葡萄糖醛酸酶,后者可分解胆汁中结合胆红素为游离胆红素,并与钙结合形成胆红素钙,促进结石的形成,认为细菌感染在胆色素类结石的形成中起着决定性作用。１９９５年Ｓｗｉｄｓｉｎｓｋｉ等〔２〕根据细菌染色体上的１６ＳｒＲＮＡ基因序列设计引物,在胆固醇类结石中特异性地扩增１６ＳｒＲＮＡ基因片段,证明多数胆固醇类结石内有细菌ＤＮＡ残留,并提出了细菌致胆固醇类结石形成的可能机理,使细菌在胆石形成中的作用研究有了新的进展。在我国胆石症的发…     

细菌与胆固醇结石

自从在胆固醇结石中扩增到细菌NDA后，细菌在胆固醇结石发病机制中的作用被重新受到重视。幽门螺杆菌在胆道疾病中的作用也是新的研究热点，本文对细菌及幽门螺杆菌在胆固醇结石形成中的作用进行文献复习。     

胆固醇结石病人与非胆石症人群胆道细菌感染状况及与免疫球蛋白相关性的对照研究

目的 明确细菌在胆固醇结石发病机制中的地位。方法 用半定量PCR结合16SrRNA序列对照法，测定38例胆囊结石病人的胆石、胆汁和胆囊黏膜细菌DNA阳性率和菌落数，并取17例非胆石病人的胆汁和胆囊黏膜作为对照。用散射比浊法测定相应胆汁和黏膜中IgA、IgG、IgM含量。结果 胆石组和非胆石组胆汁细菌DNA阳性率分别为77％和67％，菌落数分别为3．23cfu(对数值)和2．43cfu，胆囊黏膜细菌DNA阳性率分别为64％和69％，菌落数分别为3．28cfu和3．67cfu，两组间均无显著差异。两组间胆汁和黏膜IgA、IgG、IgM含量也无显著差异。菌落数与免疫球蛋白含量无相关性。结石核心和外周的细菌DNA阳性率分别为79％和82％，菌落数分别为3．19cfu和3．26cfu纯胆固醇结石细菌DNA阳性率为100％(3／3)。结石中胆固醇含量与细菌DNA阳性率无关。胆石组细菌DNA阳性者与阴性者的胆囊黏膜IgA、IgG含量均有显著差异。术前1h用抗生素者胆汁菌落数显著减少，但是黏膜菌落数无显著差异。结论 胆固醇结石病人与非胆石症人群胆道细菌感染率相似，不能仅凭结石中存在细菌来肯定细菌对胆固醇结石形成的作用。胆囊黏膜细菌与IgA、IgG含量增高有关，可能间接参与胆固醇结石形成过程。     

细菌与胆固醇结石(文献综述)

自从在胆固醇结石中扩增到细菌NDA后 ,细菌在胆固醇结石发病机制中的作用被重新受到重视。幽门螺杆菌在胆道疾病中的作用也是新的研究热点。本文对细菌及幽门螺杆菌在胆固醇结石形成中的作用进行文献复习     

色素结石形成为细菌微集落和色素粒体的复合物

以往研究提示细菌在色素胆结石的发病和临床表现中起特殊的作用,本文用电子扫描显微镜(SEM)研究胆石的形态学,测定细菌对体外色素和胆固醇结石表面的亲和性.取二组结石,一组32例新鲜结石,另一组53例干燥结石,其中取自胆囊和胆总管色素结石分别为24例和18例,胆固醇结石分别为27例和17例,混合型胆石分别为9例和11例.全组中,仅5例长期居住在东方.SEM确定有无细菌微集团固定在其表面.每一份细菌培养液作涂片,用钌红染色,测定其中糖被膜.结果: 一、结石形态学一胆固醇和色素结石的肉眼和SEM所见有明显区别: (1)色素结石的表面不     

胆固醇结石患者胆囊细菌感染的研究

目的 比较胆固醇类结石患者和非胆石症患者胆道细菌感染情况.方法 用不依赖细菌培养的半定量PCR和16SrRNA序列对照法,检测76例胆固醇类结石患者胆囊黏膜、胆汁和胆石中细菌DNA,与34例非胆石患者对照.结果 胆石组和非胆石组胆汁细菌DNA阳性率分别为77%和67%,胆囊黏膜细菌DNA阳性率分别为64%和69%,两组间差异均无统计学意义(P＞0.05).胆石组细菌种类主要为大肠杆菌、铜绿假单胞菌、金黄色葡萄球菌、不动杆菌、链球菌、鞘氨醇单胞菌、脆弱类杆菌和痤疮丙酸杆菌,胆石组菌种比非胆石组丰富,非胆石组菌种基本上均能在胆石组中找到.结论 胆固醇结石与非胆石症患者胆道细菌感染率相似,胆固醇结石中存在细菌不足以证明细菌参与胆固醇结石的形成.     

Detection of bacterial DNA from cholesterol gallstones by NP-PCR and its clinical significance

To search for bacterial DNA sequences in cholesterol gallstones with negative bacterial culture. We used nested primers polymerase chain reaction (NP-PCR) technique to amplify bacterial gene fragments were amplified in vitro from DNA extracted from cholesterol gallstones. Comparative 16S ribosomal RNA sequence analysis was used for elucidation of bacterial identification. The gallbladder gallstones of 30 patients were analyzed. Bacterial DNA was found in the stones of 26 patients. There was no difference either in cholesterol and water content or in harboring bacterial DNA of gallstones. E. coli-related DNA fragments were found in the stones of 8 patients (26.67%). Propionibacteria type DNA was found in the stones of 7 patients (23.33%). Stones of 2 patients (6.67%) harbored bacterial gene fragments with similarity of Streptococcus pyogenes. A more heterogeneous sequence collection was found in 7 patients (23.33%) and could be assigned to the multiple bacterial infections. Another stones of 2 patients (6.67%) had bacterial DNA with lower molecularweight which might be related to some unidentified bacteria. The results suggested that most cholesterol gallstones harbor bacterial DNA. It is important to determine whether these microorganisms are innocent bystanders or active participants in cholesterol gallstone formation.     

单个与多发胆固醇结石患者胆道细菌感染状况及与免疫球蛋白相关性的对照研究

目的 多发胆固醇结石患者胆囊炎症程度较高,可能与结石形成有关.对多发和单个胆固醇结石患者胆道细菌感染状况及与免疫球蛋白含量的相关性进行对照研究.方法 :用半定量PCR法测定分析38例胆囊结石患者胆石、胆汁和粘膜的细菌DNA阳性率和菌落数,并测定相应胆汁和粘膜IgA、IgG、IgM含量.结果 :单个和多发胆石组的胆汁细菌DNA阳性率分别为75.0%和73.7%,胆囊粘膜细菌DNA阳性率分别为66.7%和64.0%,结石核心的细菌DNA阳性率分别为57.1%和85.7%,结石外周细菌阳性率分别为71.4%和85.7%,两组间差异均无显著性.两组间胆汁和粘膜IgA、IgG、IgM含量差异无显著性,菌落数与免疫球蛋白含量不相关.多发胆石组粘膜细菌DNA阳性者的胆囊粘膜IgA、IgG含量高于阴性者(P<0.05).单个和多发胆石组胆汁胆固醇饱和指数(CSI)差异无显著性,各组内胆汁细菌阳性与阴性者的CSI差异也无显著性.结论 :多发和单个胆固醇结石患者胆囊细菌感染率相似,细菌感染不是多发胆固醇结石患者胆囊炎症严重的原因.多发胆固醇结石患者胆囊粘膜细菌与IgA、IgG含量增高有关,可能间接参与胆固醇结石的形成过程.     

Gallbladder bile proteins in patients with gallstones and normal controls

The concentration of total protein and 17 different proteins in gallbladder bile of patients with gallstones and normal controls were analyzed. The result showed that the amount of total protein and 11 of 17 proteins was much higher in patients with cholesterol gallstones than that of normal controls. These proteins may be involved in the nucleation of cholesterol crystal in gallbladder bile of patients with cholesterol gallstones. The nucleation activity titre (NAT) of gallbladder bile of patients with gallstones and controls was determined. The NAT of gallbladder bile of patients with gallstones was 1/25-1/100 and that of controls was 1/100-1/6400. There was a statistically significant difference of NAT between patients with cholesterol gallstones and normal controls.     

胆、肠菌群差异性与胆囊胆固醇结石形成关系

目的 基于胆囊胆固醇结石患者胆道菌群、肠道菌群差异性,探讨胆、肠菌群交互与胆囊胆固醇结石形成关系.方法 选择2020年1月至2020年6月由沧州市人民医院诊疗的胆囊胆固醇结石患者42例为研究对象,记入胆固醇结石组;胆囊炎、胆囊息肉诊治等非胆固醇结石患者40例为非胆固醇结石组.采用Illumina测序技术测序并进行细菌O...     

Crystal morphology of cholesterol monohydrate in cholesterol gallstones and gallbladder bile

To understand the pathogenesis of cholesterol gallstones from a crystallographic point of view, we investigated gallstones and precipitates in gallbladder bile by the optical method used in crystallography. First, ground thin sections of cholesterol gallstones in 28 cases were examined by a polarizing microscope. The crystalline fragments which were peeled off from gallstones by sonication were also examined in the same manner. Secondly, the cholesterol monohydrate crystals in gallbladder bile were examined by a polarizing and phase contrast microscope. The bile samples were obtained from 15 patients with cholesterol gallstones by the needle puncture of gallbladder during surgery. The results of observation were as follows. 1) The cholesterol gallstones were composed of many small plated cholesterol monohydrate crystals aggregated radially. 2) Each small plated crystal was grown by the lateral growth mechanism in bile. Spiral growth patterns were often observed on the (001) faces. 3) The cholesterol gallstones were formed by two successive mechanisms: The aggregation of cholesterol monohydrate crystals and the lateral growth of each single crystal.     

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??Study on the relationship between gallbladder cholesterol polyps and the pathogenesis of gallstones JIAO Da-hai*, HAN Tian-quan, JIANG Zhao-yan, et al. *Department of General Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Minimally Invasive Surgery Center, Shanghai 200025, China Corresponding author: WANG Ming-liang, Email: mingliangwang@hotmail.com Abstract Objective To study on the relationship between gallbladder cholesterol polyps and the pathogenesis of gallstone. Methods gallbladder stone, bile and part of gallbladder full-thickness tissue were collected from 62 patients who underwent laparoscopic cholecystectomy from April 2008 to september 2008 in our center, including 31 patients with gallbladder cholesterol polyps, 18 patients with cholesterol gallstones and 13 patients in control??6 patients with gallbladder adenoma and 7 patients with non-cholesterol gallstone ). sonography was applied to measure the gallbladder three diameter before and 1 hour after breakfast to evaluate the contracted function of gallbladder. The contents of cholesterol in gallstone and cholesterol, bile acid and phospholipid in bile were measured, and the full-thickness tissue of the gallbladder wall was done the pathological examination. Results Emptying Rate of Gallbladder in the patients with cholesterol polyps was(47.3±18.6) %, it was significantly decreased comparing with those that in the healthy people(71.7±8.1)%, P??0.01, but there ware no differences between the patients in group of cholesterol polyps and in group of gallstones??47.6±23.7??%. Cholesterol saturation index was higher in the patients with cholesterol polyps comparing with those that in control group(1.0±0.2 vs 0.6±0.3, P??0.01), but there ware no differences between the patients in group of cholesterol polyps and in group of gallstones (1.0±0.2 vs1.0±0.2, P??0.05). Gallstones and cholesterol polyps coexistence in 13 patients among 31 patients who with the gallbladder cholesterol polyps, and the incidence of gallstone is 41.9%. Conclusion the existence of gallbladder cholesterol polyps may lead to the formation of gallbladder stone.     

Triglyceride and cholesterol content in bile, blood, and gallbladder wall

Cholesterol gallstone disease is frequent in Chile compared with other countries, as is cholesterolosis of the gallbladder. The purpose of this study was to determine any differences in bile composition and cholesterol content in the gallbladder wall and serum of patients compared with findings in patients with gallstones and control subjects. In control subjects, cholesterol content of the gallbladder wall was determined in autopsy material. Patients with cholesterolosis had bile composition similar to that of patients with gallstones, with supersaturation of cholesterol in the bile. Also, these patients had increased levels of cholesterol in the gallbladder wall but normal serum cholesterol levels. These findings suggest that altered bile composition occurs in patients with cholesterolosis and gallstones.     

胆囊黏膜ABCA1的表达和胆囊胆固醇息肉发病关系的研究

目的：研究ABCA1在胆囊黏膜的表达及其表达与胆囊胆固醇息肉发病的关系。方法：收集因胆囊疾患而行腹腔镜胆囊切除术病人的胆石、胆汁、胆囊黏膜及胆囊壁全层组织共计42例,其中胆囊胆固醇息肉15例,胆囊胆固醇结石15例,对照组12例（胆囊腺瘤5例,非胆固醇胆囊结石7例）。分别测定胆石胆固醇含量、胆汁胆固醇、胆汁酸、磷脂的浓度;实时PCR定量检测胆囊黏膜ABCA1、LXRα、RXRα的mRNA表达：胆囊壁全层组织做病理切片;免疫组化显示ABCAl蛋白在胆囊黏膜的表达．结果：胆固醇息肉组胆汁胆固醇饱和指数为1．0±0．2,较对照组胆固醇饱和指数0．6±0．3明显增高,其差别有统计学显著性（P〈0．01）。免疫组化显示ABCA1在胆囊黏膜上皮细胞有明显表达。息肉组、结石组和对照组胆囊黏膜ABCAI、LXRa、RXRα mRNA相对表达量比较,各组之间差异无统计学意义。结论：胆囊黏膜ABCA1的表达可能并不是导致胆囊胆固醇息肉发病的重要原因。     

Some observations on gallstones and bile composition.

A series of 31 consecutive patients undergoing surgery for gallstone disease has been studied and the composition of the gallstones and bile from the gallbladder and common duct determined. As a result of the stone analysis by the X-ray powder diffraction method, the patients were classified according to whether their stones consisted of cholesterol, calcium salts or a mixture of the two. The mean composition of the common duct bile for the groups with cholesterol and mixed stones was just outside the micellar region of cholesterol solubility. The gallbladder bile from the cholesterol group of stone-formers was also supersaturated, but the gallblader bile from the group of mixed stone-formers was undersaturated with respect to cholesterol. None of the patients forming gallstones of calcium salts showed any abnormality in the cholesterol content of their bile.