胆囊结石联合胆汁病理检查诊断华支睾吸虫感染

目的 探讨胆囊结石联合胆汁病理检查诊断胆囊结石患者合并华支睾吸虫感染的可行性.方法 采用粪便直接涂片、胆汁离心镜检和胆囊结石病理切片镜检等方法,检查350例胆囊结石患者的粪便、胆汁和结石标本中的华支睾吸虫虫卵,分析3种方法的检出率. 结果 粪便直接涂片法、胆汁离心镜检和胆囊结石病理切片镜检华支睾吸虫虫卵的检出率分别为29.4％ (103/350)、41.5％ (145/350)和44.2％(155/350).与胆汁离心镜检法相比,粪便直接涂片法检出率最低(x2=11.05,P＜0.05),胆囊结石病理切片镜检的检出率差异无统计学意义(x2＝0.583,P＞ 0.05).胆汁、胆石联合检出率为67.7％ (237/350),明显高于其它方法(x2=48.77,P＜0.05). 结论 胆囊结石与胆汁联合病理检查虫卵可提高胆囊结石患者感染华支睾吸虫的诊断率.     

基于胆结石患者胆汁、粪便和血清标本的4种诊断华支睾吸虫感染方法的评价

目的 比较并评价粪便改良加藤厚涂片法（Kato-Katz法）、胆汁离心镜检法、血清胶体金免疫层析法（colloid gold labeled immunochromatographic,CGIA）和酶联免疫吸附试验（enzyme-linked immunosorbent assay,ELISA）检测粪便、胆汁、血清诊断华支睾吸虫感染的价值. 方法 收集92例胆结石患者的胆汁、血清和粪便,分别采用镜检法检查胆汁华支睾吸虫卵、用Kato-Katz法检查粪便华支睾吸虫卵,用CGIA法和ELISA法检测血清华支睾吸虫IgG抗体,计算并分析各检测方法的阳性率、灵敏度、特异性、阳性预测值、阴性预测值和Youden指数. 结果 Kato-Katz法联合胆汁离心镜检法共确诊感染49例,阳性率为53.3％.与之相比,Kato-Katz法、胆汁离心镜检法、CGIA和ELISA的符合率分别为80.4％、84.8％、90.2％和96.7％.Kato-Katz法、胆汁离心镜检法、CGIA和ELISA法的灵敏度分别是63.3％、93.9％、98.0％和93.9％;特异性分别为100％、100％、69.8％和86.0％;阳性预测值则分别是100％、100％、78.7％和88.5％,阴性预测值分别为70.5％、93.5％、96.8％和92.5％,Youden指数为0.63、0.94、0.68和0.80. 结论 胆汁离心镜检是胆结石手术患者诊断华支睾吸虫感染的首选方法;但对于无法获得胆汁的受检者而言,血清IgG抗体检测在排除华支睾吸虫感染、粪便Kato-Katz法在确诊感染方面有较好的临床应用价值.     

实时荧光定量PCR检测胆囊结石中华支睾吸虫DNA的研究

目的运用基于Taqman探针实时荧光定量PCR技术检测胆囊结石中的华支睾吸虫DNA。方法选取2012年3～4月份40例实施内镜取石保胆手术患者的胆囊结石,通过傅里叶红外光谱法分析结石成分,对结石研磨涂片镜检观察,并提取其DNA,运用实时荧光定量PCR技术检测华支睾吸虫基因。结果 40例患者的胆囊结石按主要成分可分为胆色素性结石(25例),胆固醇性结石(6例),混合性结石(9例)。结石涂片镜检有23例发现华支睾吸虫虫卵(阳性率57.5%),其中胆色素性结石19例,混合性结石4例。实时荧光定量PCR检测华支睾吸虫DNA阳性率为65.0%(26/40),其中包括23例镜检虫卵阳性标本及3例镜检阴性标本。结论运用实时荧光定量PCR在胆囊结石中检测出华支睾吸虫DNA,敏感性高于镜检法查虫卵。这不仅为华支睾吸虫感染诱发胆囊结石形成提供了分子生物学证据,同时也提供了一种诊断华支睾吸虫感染的有效手段,对胆囊结石病人的病因诊断、临床治疗以及术后预防复发等具有重要的指导意义。     

广西146例华支睾吸虫疑似感染者粪样虫卵和血清IgG抗体的检测分析

检测广西华支睾吸虫疑似感染者粪样华支睾吸虫/扇棘单睾吸虫虫卵和血清华支睾吸虫IgG抗体,分析二者的相关性,为华支睾吸虫感染的监测和诊断提供参考。收集2017年3-9月于广西疾病预防控制中心预防性健康体检科就诊的疑似华支睾吸虫感染者粪样,采用改良加藤厚涂片法(一粪三检)检查华支睾吸虫/扇棘单睾吸虫虫卵并计算每克粪样虫卵数(EPG)。同步收集粪检阳性患者的血液标本,采用间接ELISA法检测血清华支睾吸虫IgG抗体。分析EPG值与IgG抗体A_(450)值的相关性,并根据华支睾吸虫感染度分类标准,分析轻度、中度和重度感染者EPG值与血清IgG抗体阳性率以及IgG抗体定性结果与EPG值的关系。在146份华支睾吸虫/扇棘单睾吸虫虫卵阳性粪样中,EPG最大值为21 502,最小值为8。血清华支睾吸虫IgG抗体阳性率为84.9%(124/146),男、女性IgG抗体阳性率分别为86.0%(111/129)和13/17,差异无统计学意义(P 0.05)。感染者EPG值和IgG抗体A_(450)值呈正相关性(r_s=0.436, P 0.01)。轻度、中度和重度感染者IgG抗体阳性率分别为78.4%(69/88)、 96.0%(48/50)和7/8,差异有统计学意义(P 0.05)。华支睾吸虫血清IgG抗体阳性组和阴性组粪样几何均数(GM EPG)值分别为736和204,差异有统计学意义(P 0.01)。提示在华支睾吸虫和扇棘单睾吸虫共感染的广西,粪样虫卵镜检和血清华支睾吸虫IgG抗体检测结果呈正相关;但单独粪便虫卵检查用于华支睾吸虫监测和诊断会存在误判,应辅助IgG抗体等血清学检测。     

华支睾吸虫病流行区胆石症患者胆囊结石类型及华支睾吸虫感染情况

目的探讨华支睾吸虫病流行区胆石症患者胆囊结石类型与华支睾吸虫感染的关系。方法选取2009年5月至2012年10月在广州市南沙区第六人民医院普外科实施内镜微创取石保胆手术的胆石症患者598例。胆石样品以红外光谱法分析结石成分,判断结石类型。胆石研碎后用光镜检查华支睾吸虫虫卵,计算不同类型结石的虫卵检出率。比较不同类型的结石患者,以及碳酸钙类结石患者中华支睾吸虫虫卵阳性和阴性患者的临床特点和生化特征。选取部分虫卵阳性的碳酸钙类结石行扫描电镜观察。结果 598例胆囊结石患者中,234例(39.1%)为胆固醇类结石,133例(22.2%)为胆色素类结石,112例(18.7%)为碳酸钙类结石,86例(14.4%)为混合类结石,33例(5.5%)为其他类结石。5类胆结石华支睾吸虫卵检出率分别为6.4%(15/234)、44.3%(59/133)、59.8%(67/112)、36.0%(31/86)和30.0%(10/33)。碳酸钙类结石的虫卵检出率最高,胆固醇类结石的虫卵检出率最低。碳酸钙类结石和混合类结石患者的血清CO2结合力高于胆固醇类结石患者(P0.05),碳酸钙类结石患者胆汁CO2结合力和p H值高于其他4组结石患者(P0.05)。碳酸钙类结石患者中华支睾吸虫虫卵阳性者的血清CO2结合力、胆汁CO2结合力和p H值均高于虫卵阴性者(P0.05)。光镜和扫描电镜观察均发现,碳酸钙类结石华支睾吸虫卵与碳酸钙结晶相互黏附。结论碳酸钙类胆囊结石患者的华支睾吸虫感染率高于其他结石类型的患者。     

改良加藤厚涂片法和PCR法检测华支睾吸虫感染的 效果比较

目的　比较改良加藤厚涂片法（Kato?Katz法）和PCR法对现场人群粪便样本中华支睾吸虫感染的检出率,为华支睾吸虫感染检测方法的选择提供依据。方法　在2016年广西壮族自治区藤县人体华支睾吸虫感染流行病学调查的基础上,随机抽取133份粪便样本,-20 ℃保存。分别采用Kato?Katz法（1粪3检）和PCR法检测华支睾吸虫感染,比较两种方法对华支睾吸虫感染的检出率,采用Kappa分析评价两种检测方法结果的一致性。结果　133份人群粪便样本中,华支睾吸虫感染总检出率为77.44%（103/133）,其中Kato?Katz法检出率为57.14%（76/133）,PCR法检出率为70.68%（93/133）, PCR法检出率显著高于Kato?Katz法（[χ2] = 26.15,P <0.01）。76份Kato?Katz法阳性粪便样本中,88.16%（67/76）的样本PCR法扩增阳性;57份Kato?Katz法阴性样本中,47.37%（27/57）的样本PCR法扩增阳性。PCR法对每g粪便虫卵数（EPG）> 1 000的粪便样本中华支睾吸虫感染的检出率（94.7%,18/19）高于EPG < 1 000的样本（85.96%,49/57）,但差异无统计学意义([χ2] = 1.05,P = 0.436)。两种方法检出率一致性检验结果一般（Kappa = 0.73）。结论　PCR法对人群华支睾吸虫感染的检出率显著高于Kato?Katz法。建议在华支睾吸虫感染度较轻的地区,采用Kato?Katz法结合PCR法进行检测,以提高检出率。     

广西藤县人群华支睾吸虫感染调查及其分子鉴定

目的了解广西藤县人群华支睾吸虫的感染状况及其分子鉴定,为当地华支睾吸虫病的防控提供理论依据。方法2016年11月,在广西藤县采用分层整群随机抽样抽取天平镇和金鸡镇各1个自然村作为调查点。收集3岁以上常驻居民的粪便,用改良加藤厚涂片法(一粪三检)检测华支睾吸虫虫卵并计数。通过问卷调查收集被调查者社会人口学等相关信息。从镜检阳性样本中随机选取部分新鲜粪样提取DNA, PCR扩增华支睾吸虫内转录间隔区2 (ITS2),测序后在NCBI核酸数据库作比对分析。结果共调查400人,检出华支睾吸虫虫卵阳性208人,感染检出率为52.0%(208/400),感染者以轻度感染为主(69.2%, 144/208)。天平镇华支睾吸虫感染检出率为66.5%(133/200),高于金鸡镇的37.5%(75/200)(χ2=33.69, P 0.01)。男性感染检出率为58.7%(131/223),高于女性的43.5%(77/177)(χ2=9.18, P 0.01); 30～39岁组感染检出率最高,为63.8%(60/94);高中及以上学历人群感染检出率最高,为68.0%(17/25)。不同年龄组、教育程度人群感染检出率差异有统计学意义(χ2=37.1, 23.02; P 0.01)。PCR扩增出156 bp的华支睾吸虫特异性条带,其序列与越南岘港分离株ITS2 (GenBank登录号:MF319655)的一致性为100%。结论广西藤县人群华支睾吸虫感染检出率为52.0%,属于超重流行区;男性、青壮年是华支睾吸虫高感染人群。广西藤县的华支睾吸虫ITS2序列与越南岘港分离株的基因型一致。     

2016—2020年福建省浦城县华支睾吸虫病国家监测点人群感染情况分析

2016—2020年根据《全国肝吸虫病和土源性线虫病监测方案》要求,将浦城县按地理方位划分东、南、西、北、中等5个片区,每个片区随机抽取1个乡镇的1个村作为人群华支睾吸虫感染情况的国家监测点。每个村随机调查200人以上,采集人群粪样采用改良加藤厚涂片法检测华支睾吸虫虫卵(一粪二检);根据村庄鱼塘溪渠分布情况,每年度采集各类淡水鱼类100尾,采用压片镜检法检测鱼鳃、鱼鳍及尾鳍部肌肉的华支睾吸虫囊蚴;各年度收集野外犬、猫粪样共25份,以水洗沉淀法检测华支睾吸虫虫卵;在监测点受检人群中随机抽查150人以上进行华支睾吸虫病防治知识、态度和行为等内容的问卷调查。2016—2020年共调查5 553人,吸虫总感染率为0.79%(44/5 553)。各年度感染率分别为0.60%、 1.02%、 1.16%、 0.97%和0.24%。其中,水洗沉淀检出华支睾吸虫感染者26人,感染率为0.47%(26/5 553);检出东方次睾吸虫感染者18人,感染率为0.32%(18/5 553)。检测3种淡水鱼类共639尾,感染囊蚴鱼类472尾,感染率为73.86%。其中,麦穗鱼的感染率2016年最高,达97.33%(73/75)。检测野外动物粪样共129份,检出华支睾吸虫卵5份,东方次睾吸虫卵2份,检出率分别为3.88%和1.55%。居民有效问卷共752份,总知晓率为37.10%(279/752)。男性与女性知晓率差异无统计学意义(χ2=0.73, P 0.05)。浦城县人群华支睾吸虫感染率低,居民对华支睾吸虫病知识的知晓率低。当地淡水鱼类特别是麦穗鱼的囊蚴感染率较高,应加强宣传教育与防治工作。     

对流免疫电泳检测粪内华支睾吸虫抗原诊断华支睾吸虫病的试验研究

华支睾吸虫卵甚小,粪检时极易漏检。十二指肠及胆汁引流等方法,虽可提高检查效果,然操作不便,难于推广。血清学方法虽可提高检出率,但不易区分既往感染和现症病人。因此,我们初步探讨了对流免疫电泳法检测华支睾吸虫病人粪便内抗原的诊断价值。一、抗华支睾吸虫兔免疫球蛋白制备:     

华支睾吸虫病免疫诊断的近年进展

华支睾吸虫病的诊断,主要靠粪检虫卵。但在感染度较轻的情况下,常因粪便内虫卵少,加以虫卵很小,粪样镜检易致漏诊。近年来,随着酶、同位素、胶体金等标记技术和新方法的应用,华支睾吸虫病的免疫诊断获得较大进展,几种检测华支睾吸虫抗体或抗原的新方法具有较高的敏感性和特异性。现将这几种方法作一简要综述。     

集卵薄涂片法检测牛粪中血吸虫卵含量的研究

本文比较了薄涂片法和透明法的虫卵回收率。结果表明,在三种虫卵梯度,薄涂片法的虫卵回收率,分别为透明法的18、4.2和3.5倍,前者的变异系数明显低于后者;现场检查34头孵化阳性黄牛的结果也证实薄涂片法优于透明法。薄涂片法检查半量粪渣(制6张涂片)的虫卵估计值和标准差与全量粪渣结果相似,可以代替全量粪渣检查。在感染度较高的地区,可以抽样检查1/4粪渣,制3张涂片检查,从而减少工作量。     

华支睾吸虫感染是胆囊结石患者合并壁问结石的危险因素

目的探讨华支睾吸虫感染与胆囊结石患者合并壁间结石的关系。方法回顾性分析2011年1～12月间连续340例实施硬镜取石保胆手术的胆囊结石病人合并胆囊壁间结石情况,及其与华支睾吸虫感染、年龄、性别、术前血糖、血脂水平、乙肝病毒感染、胆囊结石的数量和大小的关系。结果①340例胆囊结石患者中发现合并壁间结石79例（23．2％）检出华支睾吸虫感染阳性153例（45．0％）。②华支睾吸虫感染阳性患者合并壁间结石49例（49／153,32．0％）;感染阴性患者合并壁间结石30例（30／187,16．0％）,感染阳性者合并结石率较高（P=0．01）;79例合并壁间结石的病人中,47例壁间结石内发现华支睾吸虫卵（59．5％）。③Logistic回归分析显示,合并壁间结石的发生,与年龄、性别、术前血糖、血脂水平、乙肝病毒感染、胆囊结石的数量和大小等因素关系不大,而与华支睾吸虫感染有关（OR-2．29;95％CI=1．35～3．86,P=0．002）。结论华支睾吸虫感染是胆囊结石患者合并砖间结石的危险因素．。     

Detection of Helicobacter pylori by polymerase reaction in bile samples from gallbladder and bile stones

Some papers report helicobacter pylori existence in bile from surgical specimens obtained during gallbladder or bile ducts surgery. The aim of this work was search by PCR, H. Pylori presence in bile specimens from patients suffering of gallbladder stones or by bile ducts stones. Bile samples were obtained by gallbladder punction during cholecystectomy in 26 patients, 19 of them with gallbladder stones and 7 also with gallbladder stones and bile duct stones. Age ranged from 22-69 years old, median 49.6 years old. Samples were sent to specialized biomolecular laboratory to perform PCR techniques. Two of 26 patients (7.6%) had positive reaction for the presence of DNA of H. Pylori in bile samples. Our research suggest that DNA of H. Pylori can be founded in bile samples patients with gallbladders and duct stones in Argentina.     

Clonorcis sinensis eggs are associated with calcium carbonate gallbladder stones

Calcium carbonate gallbladder stones were easily neglected because they were previously reported as a rare stone type in adults. The aim of this study was to investigate the relationship between calcium carbonate stones and Clonorchis sinensis infection. A total of 598 gallbladder stones were studied. The stone types were identified by FTIR spectroscopy. The C. sinensis eggs and DNA were detected by microscopic examination and real-time fluorescent PCR respectively. And then, some egg-positive stones were randomly selected for further SEM examination. Corresponding clinical characteristics of patients with different types of stones were also statistically analyzed. The detection rate of C. sinensis eggs in calcium carbonate stone, pigment stone, mixed stone and cholesterol stone types, as well as other stone types was 60%, 44%, 36%, 6% and 30%, respectively, which was highest in calcium carbonate stone yet lowest in cholesterol stone. A total of 182 stones were egg-positive, 67 (37%) of which were calcium carbonate stones. The C. sinensis eggs were found adherent to calcium carbonate crystals by both light microscopy and scanning electron microscopy. Patients with calcium carbonate stones were mainly male between the ages of 30 and 60, the CO₂ combining power of patients with calcium carbonate stones were higher than those with cholesterol stones. Calcium carbonate gallbladder stones are not rare, the formation of which may be associated with C. sinensis infection.     

Relationship between intraduodenal peri-ampullary diverticulum and biliary disease in 178 patients undergoing ERCP

BACKGROUND:The presence of intraduodenal peri-ampullary diverticulum is often observed during upper digestive tract barium meal studies and endoscopic retrograde cholangiopancreatography(ERCP).A few papers in China and overseas reported that the diverticulum had something to do with the incidence of cholelithiasis. This study was undertaken to further test this notion and ascertain the relationship between intraduodenal peri- ampullary diverticulum and biliary disease,especially the formation of bile duct pigment stones. METHODS:A total of 178 patients who had undergone ERCP or endoscopic sphincterotomy(EST)were studied retrospectively.They were divided into 6 groups according to the category of biliary disease,and the incidence rates of intraduodenal peri-ampullary diverticulum were calculated. RESULTS:There were 44 patients with intraduodenal peri- ampullary diverticulum in 81 patients with primary bile duct pigment stones(54.32%),4 in 8 patients with bile duct stones and gallbladder stones(50%),7 in 33 patients with bile duct stones secondary to gallbladder stones(21.21%), 3 in 21 patients with inflammation and stricture of the end of the bile duct and papilla(14.29%),1 in 22 patients with carcinoma of the end of the bile duct and papilla(4.54%), and 5 in 13 patients with post-cholecystectomy syndrome or sphincter of Oddi dysfunction(38.46%). CONCLUSIONS:The incidence rate of intraduodenal peri- ampullary diverticulum in patients with primary bileduct pigment stones is higher than that in patients with bile duct stones secondary to gallbladder stones,patients with inflammation and stricture of the end of the bile duct and papilla,and patients with carcinoma of the end of the bile duct and papilla.These findings indicate that the anatomical abnormalities and malfunction of the sphincter of Oddi play an important role in the formation of bile duct pigment stones.     

Sensitivity and specificity of microscopic examination of gallbladder bile for gallstone recognition and identification

During cholecystectomy, gallbladder bile and gallstones were obtained from 77 patients and gallbladder bile was obtained from 39 patients free of stones (11 patients had biliary stenosis). According to their chemical composition, gallstones were classified as cholesterol (n = 46) or pigment (n = 31) stones. In patients with gallstones (a) cholesterol crystals better helped to identify cholesterol gallstones (sensitivity, 87%; specificity, 97%; positive predictive value, 97%) than did an abnormal cholesterol saturation index of bile (sensitivity, 93%; specificity, 48%; positive predictive value, 73%); (b) the presence of cholesterol crystals was significantly related to the cholesterol content of gallstones and the bile cholesterol saturation index; and (c) bilirubinate crystals, when present alone (without cholesterol crystals), were good predictors of pigment gallstones (sensitivity, 71%; specificity, 93%; positive predictive value, 88%). In the absence of stones, bilirubinate crystals were present in 9 of 28 patients without biliary stenosis (4 with alcoholic cirrhosis and 2 with alcoholic pancreatitis) and 8 of 11 patients with biliary stenosis. In the absence of stones, cholesterol crystals were present in 2 of 28 patients without biliary stenosis and in 4 of 11 patients with biliary stenosis, suggesting that bile stasis can induce cholesterol crystal formation.     

Treatment of gallstones with piezoelectric lithotripsy and oral bile acids. A multicenter study

The efficacy of the combination of piezoelectric lithotripsy and oral bile acids in the treatment of gallbladder stones was assessed. Three hundred and sixty-three patients with symptomatic radiolucent gallstones in functioning gallbladder were treated in five medical centers using the same protocol with the EDAP LT 01 lithotripter. No anesthesia, analgesia or sedation was used. After one session of lithotripsy, fragmentation was observed in 89% of the patients, and satisfactory fragmentation (fragments less than or equal to 5 mm) in 29%. The satisfactory fragmentation rate was higher in patients with solitary stones less than or equal to 20 mm than in patients with solitary stones 21-35 mm or multiple stones (p less than 0.001). After multiple sessions (mean 1.6 session/patient, range 1-5) the overall rate of satisfactory fragmentation was 50%. After 12 months on oral bile acid therapy, complete clearance of the gallbladder was observed in 69% of patients with solitary stones less than or equal to 20 mm, 25% of patients with solitary stones 21-35 mm and 37% of patients with multiple stones. No complication was observed during the lithotripsy. During follow-up under bile acid therapy, there were five complications (1.4%): four patients had acute cholecystitis and one had mild, self-limited pancreatitis. We conclude that piezoelectric lithotripsy with the EDAP lithotripter is a safe and effective treatment which can be performed in outpatients. Satisfactory fragmentation and rapid disappearance of stones are obtained mainly in patients with solitary stones less than or equal to 20 mm.     

一步法和分步法治疗胆囊结石合并胆总管结石患者疗效及并发症分析

目的 探讨一步法与分步法手术治疗胆囊结石合并胆总管结石患者的疗效。方法 2016年1月～2019年6月我院诊治的胆囊结石合并胆总管结石患者152例,其中87例接受一步法手术,即行腹腔镜胆囊切除(LC)和胆总管探查取石(LCBDE)术,65例接受分步法手术,即行内镜逆行胰胆管造影/乳头括约肌切开(ERCP/EST)和LC术,比较两组疗效。结果 一步法组总手术时间为(156.3±42.6)min,术后置管率为98.9%,显著长于或高于分步法组【(124.4±31.4)min和15.4%,P<0.05】,而住院日为(14.4±3.9)d,住院费用为(27.9±7.0)千元,均显著短于或低于分步法组【(18.7±4.2)d和(31.24±5.8)千元,P<0.05】;一步法组手术成功率为95.4%、结石清除率为93.1%,分步法组分别为100.0%和96.9%,差异均无统计学意义(P>0.05);一步法组术后反流性胆管炎发生率0.0%,显著低于分步法组的7.7%(P<0.05),但两组并发症发生率比较差异无统计学意义(19.5%对 32.3%,P>0.05)。结论 一步法和分步法手术均可应用于治疗胆囊结石合并胆总管结石患者,但在合适的患者采取一步法手术可能更迅速地解除病痛,疗效好。