检测华支睾吸虫特异性IgG4生物素-亲和素复合酶联免疫吸附法的建立及检测效能分析

目的探讨亲和素-生物素复合酶联免疫吸附法(ABC-ELISA)检测特异性Ig G4抗体用于诊断华支睾吸虫病的效果。方法建立检测华支睾吸虫特异性抗体Ig G4的ABC-ELISA法(Ig G4-ABC-ELISA),以此方法检测华支睾吸虫病、日本血吸虫病、卫氏并殖吸虫病、弓形虫病、棘球蚴病、囊尾蚴病和曼氏裂头蚴病患者血清样本。以Ig G4-ELISA和Ig GELISA法为对照,比较3种方法用于诊断华支睾吸虫病的敏感性、特异性、阳性预测值、阴性预测值及诊断效率。结果成功建立了用于检测华支睾吸虫特异性抗体的Ig G4-ABC-ELISA法,用此方法检测华支睾吸虫病患者血清特异性抗体Ig G4的敏感性为90.0%,特异性为98.2%,阳性预测值为93.8%,阴性预测值为97.0%,诊断效率为96.3%;Ig G4-ELISA法检测华支睾吸虫病患者血清特异性抗体敏感性为86.0%,特异性为98.2%,阳性预测值为93.5%,阴性预测值为95.9%,诊断效率为95.4%;Ig G-ELISA法检测华支睾吸虫病患者血清特异性抗体Ig G的敏感性为94.0%,特异性为88.1%,阳性预测值为70.1%,阴性预测值为98.0%,诊断效率为89.4%。Ig G4-ABC-ELISA法检测华支睾吸虫病患者血清特异性抗体的敏感性高于Ig G4-ELISA法(P0.05),特异性高于Ig G-ELISA法(P0.05)。结论 Ig G4-ABC-ELISA法检测华支睾吸虫特异性抗体IgG4具有高度敏感性与特异性,在华支睾吸虫病诊断中具有较好应用价值。     

华支睾吸虫病的超声诊断价值探讨

目的对比分析850例华支睾肝吸虫病患者与未查出华支睾吸虫病声像图表现作对比,探讨超声对本病的诊断价值。方法对已确诊的850例华支睾肝吸虫病患者进行肝、胆、脾、胰超声检查,观察其声像图表现与对照组进行比较。结果华支睾肝吸虫病声像图上主要表现为肝左叶增大;肝内胆管胆管壁(二级分支)增厚、轻度扩张、回声增强、以肝左叶胆管明显;另外可出现胆囊和肝实质改变。结论华支睾肝吸虫病具有特征性的声像图表现,超声检查操作简便,可作为普查手段,对本病做出早期诊断。     

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.     

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。     

重组抗原应用于SPG-ELISA检测华支睾吸虫循环抗体的效果评价

目的评价重组抗原用于SPG-ELISA诊断华支睾吸虫感染的价值。方法分别使用重组华支睾吸虫半胱氨酸蛋白酶（Cs-CysB）与华支睾吸虫成虫可溶性抗原（CAA）,以SPG-ELISA检测华支睾吸虫病患者血清特异性IgG,比较两种抗原包被SPG-ELISA方法的敏感性与特异性。结果检测50份华支睾吸虫感染者血清,重组Cs-CysB与CAA包被的SPG-ELISA阳性率均为94.00%,健康人血清50例均为阴性;检测日本血吸虫病血清20份、并殖吸虫病血清20份、囊虫病血清10份,交叉反应率分别为5.00%、10.00%、20.00%和5.00%、0、20.00%。结论以重组Cs-CysB为包被抗原用SPG-ELISA检测血清华支睾吸虫抗体具有较高的敏感性及特异性,检测效果与CAA包被的SPG-ELISA相当。     

华支睾吸虫半胱氨酸蛋白酶的基因表达与在ELISA诊断上的应用研究

目的探讨重组华支睾吸虫半胱氨酸蛋白酶在华支睾吸虫病血清学诊断上的应用价值. 方法以成虫cDNA为模板PCR扩增GenBank中一个华支睾吸虫半胱氨酸蛋白酶(No. AF093242,简称CysB)的部分基因片段,亚克隆到pTrcHis表达载体,转化入大肠埃希菌DH5α.表达出的重组蛋白经亲合层析纯化后用于ELISA检测华支睾吸虫及其他寄生虫感染血清,评价其诊断应用价值. 结果成功克隆与表达CysB基因片段,纯化后的重组蛋白用于华支睾吸虫病诊断的敏感性达96.0%(48/50),与其他寄生虫病的总交叉反应为3.8%(1/26),而对比实验中可溶性粗抗原(CAA)用于检测的敏感性为88.0%(44/50),与其他寄生虫感染血清无交叉反应. 结论重组华支睾吸虫半胱氨酸蛋白酶(CysB)用于ELISA诊断技术具有较高的敏感性及特异性,有希望成为可溶性粗抗原的替代之一.     

华支睾吸虫感染的超声诊断分析

【提要】 回顾性分析临床确诊有华支睾吸虫感染的214例患者治疗前后的超声表现，结果感染后的超声表现以胆囊及胆管病变为主，胆囊内出现的絮状弱回声漂浮物具有特征性，治疗后该漂浮物消失。绝大多数患者有胆管回声改变，表现为胆管壁增厚、毛糙，回声增强，治疗后恢复较慢。超声对该病的诊断有应用价值。     

广州珠三角地区家猫自然感染华支睾吸虫的调查及豚鼠模型的构建北大核心CSCD

目的了解广州珠三角地区家猫自然感染华支睾吸虫状况,观察豚鼠感染华支睾吸虫后肝组织病理变化。方法解剖当地家猫,取肝脏,检查华支睾吸虫感染情况,阳性肝脏收集成虫,并取肝组织制作切片;采集阳性鱼,分离华支睾吸虫囊蚴,经口感染豚鼠,60个囊蚴/只,60d后解剖豚鼠,检查肝脏,收集成虫,取病变肝组织制作切片,作病理检查。实验设未感染对照组。结果当地猫华支睾吸虫自然感染率为41.47%(214/516);感染豚鼠华支睾吸虫成虫回收率50.83%(305/600)。与对照组比较,实验组豚鼠肝脏肿大,边缘呈球状隆起,部分肝叶表面可见水泡状凸起病变,水泡内液清亮;病变组织横切面可见胆管管壁增厚,管腔内有华支睾吸虫成虫寄生;镜下可见华支睾吸虫虫卵切面,周围大量炎症细胞浸润,胆管管壁增厚,管周纤维组织增生伴胆管扩张,肝小叶结构破坏。结论广州珠三角地区家猫华支睾吸虫感染率较高。豚鼠是华支睾吸虫合适的终末宿主,且肝脏病变明显。     

胆囊结石研碎镜检法对华支睾吸虫感染检出率的研究

收集2010年1~6月于广州市番禺区第二人民医院普外科实施内镜微创取石保胆术的179例患者的血清、粪便、胆汁和胆囊结石标本,分别采用胶体金免疫层析法检测华支睾吸虫IgG抗体,粪便直接涂片法、胆汁离心沉渣镜检法和胆囊结石研碎镜检法检查华支睾吸虫虫卵,并进行比较分析。结果显示,胶体金免疫层析法检测华支睾吸虫IgG抗体的阳性率为51.4%(92/179),粪便直接涂片法、胆汁离心沉渣镜检法和胆囊结石研碎镜检法华支睾吸虫卵阳性检出率分别为30.7%(55/179)、44.7%(80/179)和69.8%(125/179),粪便直接涂片法检出率最低,胆囊结石研碎镜检法检出率最高(均P<0.05)。其中粪便直接涂片法和胆汁离心沉渣镜检法检测结果为阳性的患者,其结石研碎镜检法的检测结果也均为阳性。     

胆管梗阻合并华支睾吸虫感染患者临床特征分析

目的　分析胆管梗阻合并华支睾吸虫感染患者临床特征,为该病临床诊疗提供参考。方法　收集2012年10月—2019年9月在哈尔滨医科大学附属第二医院就诊的45例胆管梗阻合并华支睾吸虫感染患者,并随机选取同期就诊的45例单纯胆管梗阻患者作为对照。采用单因素分析华支睾吸虫感染相关流行病学特征、临床表现、实验室检查结果及影像学表现,并将有统计学意义的单因素作为自变量进行多因素logistic回归分析。结果　与单纯胆管梗阻患者相比,胆管梗阻合并华支睾吸虫感染患者中男性占比更高（91.11% vs. 46.67%;[χ2] = 20.737,P < 0.01）、农村病例占比更高（62.22% vs. 22.22%;[χ2] = 14.757,P < 0.01）,主要临床症状为黄疸（45例,占100%）和腹痛（40例,占88.89%）。多因素logistic回归分析显示,男性[比值比（OR） = 10.717,95%可信区间（CI）: （2.571,44.662）]和有饮酒史[OR = 4.474,95% CI: （1.019,19.642）]是胆管梗阻合并华支睾吸虫感染危险因素;居住地为城市[OR = 0.128,95% CI: （0.038,0.435）]为保护因素。实验室检查发现,胆管梗阻合并华支睾吸虫感染患者较单纯胆管梗阻患者总胆红素和直接胆红素水平降低,而间接胆红素水平升高,差异均有统计学意义（Z = -2.566、-3.454和-3.821,P均< 0.05）。影像学检查发现,胆管梗阻合并华支睾吸虫感染患者胆管管壁增厚、肝内胆管扩张。结论　胆管梗阻合并华支睾吸虫感染患者临床表现具有特征性,在临床工作中可结合患者流行病学特征、临床症状、实验室及影像学检查结果等进行综合鉴别诊断。     

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.     

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.     

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.     

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。     

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.     

Angiographic diagnosis of the degree of serosal invasion of carcinoma of the colon

Angiography using Prostaglandin El^® was performed on 38 patients with carcinoma of the colon in order to diagnose the degree of serosal cancer invasion. The findings at angiography were classified into four groups:1) AG-S3, abnormal change (irregularity and/or encasement) up to marginal vessels; 2) AG-S2, abnormality up to vasa recta; 3) AG-S1, abnormality of penetrating branches of vasa recta within the wall of the colon; and 4) AG-S0, no distinct findings of abovementioned vessels. These angiographic findings were compared with both macroscopic and microscopic serosal cancer invasion. Angiographic diagnosis is in accord with the macroscopic findings in 84.2 percent of cases. Angiographic diagnosis is in accord with the microscopic findings in 32.4 percent of cases. Macroscopic findings confirm the angiographic diagnosis precisely but the conflict with microscopic findings should not be overlooked. This may be the result of inflammatory change, adhesion, and fibrosis around carcinoma of the colon.     

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.