胶体金免疫层析法与双单克隆抗体夹心ELISA检测鼠疫F1抗原的比较

目的比较胶体金免疫层析法（GICA）和双单克隆抗体夹心酶联免疫吸附试验（DMcAbS-ELISA）检测鼠疫F1抗原的敏感性和特异性。方法用GICA试纸条和DMcAbS-ELISA平行检测308份强毒鼠疫耶尔森菌感染鼠脏器标本和327份对照鼠标本,用RIHA微量法同时检测作为参照。结果327份对照鼠鼠疫细菌学检验及GICA,DMcAbS-ELISA和RIHA的F1抗原检测均为阴性,GICA,DMcAbS-ELISA和RIHA在108cfu/mL水平上未发现与近缘假结核耶尔森菌有交叉反应;308只感染鼠样本细菌培养阳性284只,24只样本未分离到鼠疫菌;F1抗原检测GICA阳性287份,DM-cAbS-ELISA阳性280份,RIHA阳性276份,GICA阳性检出率最高93.19%,与DMcAbS-ELISA和RIHA比较,差异均有统计学意义（χ^2=5.14,9.09;P=0.016,0.001）;GICA与DMcAbS-ELISA符合率97.73%（Kappa=0.845）;GICA和DMcAbS-ELISA与RIHA符合率分别是96.43%和98.70%（Kappa=0.774,0.926）;284份细菌培养阳性鼠标本F1抗原检测GICA阳性率是98.59%,高于DMcAbS-ELISA和RIHA,差异有统计学意义（χ^2=4.17,5.14;P=0.031,0.016）;24份细菌培养阴性实验鼠标本F1抗原检测：GICA检出7份阳性,阳性率29.17%,DMcAbS-ELISA检出6份阳性,阳性率25%,RIHA检出3份阳性,阳性率12.5%,GICA高于DMcAbS-ELISA和RIHA,但差异无统计学意义（χ^2=2.25,0;P=0.125,1.000）。结论GICA检测鼠疫F1抗原敏感特异,快速简便,优于DMcAbS-ELISA和RIHA,是鼠疫快速诊断中有应用价值的检测技术。     

2015─2017年新疆青河县山区动物鼠疫血清学和病原学调查

目的通过鼠疫病原学和血清学的实验检测结果,了解新疆青河县山区存在鼠疫自然疫源地的潜在状况。方法采用鼠疫"四步检验"法分离青河县山区捕获的长尾黄鼠及灰旱獭脏器、自毙动物脏器及骨骼和寄生蚤中的鼠疫菌;间接血凝试验(HA)和酶联免疫吸附试验(ELISA)检测动物脏器和寄生蚤悬液中的鼠疫F1抗原;长尾黄鼠、灰旱獭以及牧犬血清中的鼠疫F1抗体。结果经鼠疫"四步检验"和反向间接血凝(RIHA)检验长尾黄鼠脏器552份、灰旱獭脏器4份、自毙长尾黄鼠和灰旱獭脏器及骨骼13份、长尾黄鼠体外寄生蚤583只,均未检出鼠疫菌及F1抗原;检测长尾黄鼠血清552份,IHA法检出阳性1份、滴度1∶2~9,ELISA法检出阳性3份、平均滴度1∶2~8;检测牧犬血清129份,IHA法检出阳性6份、平均滴度1∶2^(3.83),ELISA法检出阳性8份、平均滴度1:26.75;其中牧犬鼠疫F1抗体阳性标本中检出鼠疫IgM抗体阳性2份,平均滴度1∶2^(5.5)。结论新疆青河县山区啮齿类动物间有鼠疫流行迹象,流行区域为花海子地区,存在潜在鼠疫自然疫源地,建议对该区域开展进一步鼠疫疫源性调查。     

鼠疫检验新技术在乌鲁木齐县灰旱獭鼠疫疫源地中的应用

目的比较分析酶联免疫吸附试验(ELISA)、红细胞凝集试验(IHA和RIHA)、胶体金纸上层析试验(GICA)3种免疫学检测技术的敏感性、特异性和稳定性。方法按照《鼠疫检测新技术在灰旱獭鼠疫监测和应急处置中的应用》课题设计要求,以Spss 13.0对2007—2015年乌鲁木齐县鼠疫监测数据整理并进行分析。结果 2007—2015年检测血清样本3 260份,其中灰旱獭血清2 753份、牧犬血清507份,ELISA检出阳性36份、阳性率1.10%,IHA检出阳性13份、阳性率0.40%,两种方法检出阳性率差异有统计学意义(χ~2=10.877 7,P0.01),检出鼠疫抗体滴度差异无统计学意义(t=0.036 6,P0.01);共检测以自毙灰旱獭脏器及骨骼为主的各种自毙动物材料322份,检出鼠疫F1抗原分别为ELISA8份、GICA 8份和RIHA 6份,阳性率分别为2.48%、2.48%和1.86%,四步检验法分离出鼠疫菌2株、检菌阳性率0.62%,ELISA与RIHA检出阳性率(χ~2=0.292 1)和检菌率(χ~2=3.465 2)差异无统计学意义(P0.01),RIHA与四步法检菌率差异亦无统计学意义(χ~2=1.925 5,P0.01);检测自毙动物脏器标本鼠疫F1抗原ELISA和GICA阳性率一致,二者略高于RIHA和检菌,差异无统计学意义(χ~2=0.292 1,P0.01)。结论在鼠疫监测和应对突发鼠疫疫情处置工作中,3种方法各有其优缺点,联合使用才能发挥各自特长,依据不同情况采用不同的方法,可提高鼠疫监测质量和快速应对鼠疫突发事件能力。     

酶免疫染色技术快速检测鼠疫耶尔森菌

目的研究酶免疫染色技术检测鼠疫耶尔森菌(鼠疫菌)的敏感性及特异性,建立鼠疫免疫快速检测技术。方法用酶免疫染色技术对细菌标本和动物标本检测鼠疫菌,动物标本同时用细菌学、RIHA、RGICA和ELISA法平行检测鼠疫菌。结果细菌标本酶免疫染色法检测结果与实际相符,酶免疫染色技术检测鼠疫菌的灵敏性为1 000个/ml;动物标本酶免疫染色法检测鼠疫菌结果和细菌学、RIHA、RGICA和ELISA一致。结论酶免疫染色技术检测鼠疫菌敏感、特异、简便、快速,可用于鼠疫检测工作。     

胶体金免疫层析法检测鼠疫F1抗原的研究

目的 研究胶体金免疫层析试纸条检测鼠疫F1抗原的敏感性和特异性.方法通过鼠疫F1抗原单克隆抗体(F1MAb)捕捉F1抗原的胶体金免疫层析(GICA)试纸条,检测308只强毒鼠疫耶尔森菌(鼠疫菌)感染鼠脏器标本和225份对照鼠(达乌尔黄鼠217只,小白鼠5只,豚鼠3只)标本,同时用鼠疫反向间接血球凝集试验(RIHA)微量法和细菌培养做平行检测.结果 225只对照鼠鼠疫细菌学检验及GICA和RIHA的F1抗原检测均为阴性,GICA和RIHA在1×108 cfu/ml水平上未发现与近缘假结核耶尔森菌有交叉反应.GICA、RIHA对鼠疫菌检测灵敏性为2.5×105、2.0×105cfu/ml;检测F1抗原灵敏性为1、10 μg/L.GICA与细菌学检验符合率为97.94%(522/533),Kappa值=0.959,阳性检出率组间比较,差异无统计学意义(x2=0.36,P＞0.05);与RIHA符合率为97.94%(522/533),Kappa值=0.959,阳性检出率组间比较差异有统计学意义(x2=9.09,P＜0.05).308只实验鼠样本细菌培养阳性284只,284份细菌培养阳性标本中,GICA有280份检测阳性,阳性率为98.59%,高于RIHA[阳性率为96.13%(522/533)],两组比较差异有统计学意义(x2=5.14;P＜0.05).GICA的敏感度为98.59%(280/284),特异度为97.19%(242/249),阳性预测值为97.56%(280/287),阴性预测值为98.37%(242/246),Youden指数为0.9578.结论 GICA检测鼠疫F1抗原敏感特异,快速简便,是鼠疫早期快速诊断中有应用价值的检测技术.     

不同方法检测鼠疫动物脏器的研究

应用双重式聚合酶反应（Pla-F1-PCR)、常规细菌培养和反相血凝试验（RIHA），对9只实验感染鼠疫的动物脏器（豚鼠6份，小白鼠3份），在4℃条件下保存不同时间后进行同步检测。结果表明，保存90d和180d后，PCR与RIHA均为阳性，而细菌培养阳性率明显降低，PCR和RIHA法敏感性高于常规培养。提示对鼠疫或疑似鼠疫材料的检测除常规培养外应做PCR和RIHA，以防漏诊。     

鼠疫反向间接血凝试验与鼠疫细菌培养结果的对比分析

目的对比鼠疫反向间接血凝试验(RIHA)与鼠疫细菌培养的检菌结果,选择灵敏、简便、快速的鼠疫诊断方法。方法用鼠疫菌感染小鼠、大耳白兔、豚鼠,动物死亡后取肝、脾脏;大耳白兔死亡后,在室温(18～22℃)下自然腐败,并于第0、15、60天取脾脏;取人感染鼠疫尸体的肺脏;将上述样品同时进行RIHA和细菌培养,计算鼠疫菌阳性率,比较两种方法的检菌效果。结果动物和人被鼠疫菌感染后,鼠疫RIHA的阳性率(100％)明显高于细菌培养(87％);对腐败材料鼠疫菌的检测,RIHA诊断效果明显优越于细菌培养,尤其在菌培养阴性的情况下,仍能检查出鼠疫菌抗原(抗体滴度为1:102 400)。结论RIHA在鼠疫诊断中,具有敏感、简便和快速的特点,有助于及时确定疫源的存在和疫情的判定,在实际工作中便于应用和推广,希望早日成为鼠疫实验室诊断标准。     

检测鼠疫F1抗原的三种免疫学方法比较

目的 比较三种检测鼠疫F1抗原方法用于诊断鼠疫的效果.方法在鼠疫疫源地,收集野生动物的血、肝、脾、淋巴组织标本,酶联免疫吸附试验(ELISA)、反向间接血凝试验(RIHA)、胶体金纸上色谱试验(GICA)三种方法检测鼠疫F1抗原.结果检测鼠疫疫源地内自毙和捕获野生动物脏器浸液标本414份,ELISA和GICA检出的阳性标本相同,阳性率均为5.31%(22/414),阳性符合率和阴性符合率均为100%.RIHA初试2孔以上凝集186份,复试仅18份阳性,非特异性凝集率高达40.57%(168/414),阳性率4.35%(18/414).RIHA与GICA、ELISA的阳性符合率均为81.82%(18/22),阴性符合率均为98.99%(392/396);阳性标本的平均滴度,ELISA为1:29.95,RIHA为1:28.68,组间比较差异有统计学意义(t=4.379,P＜0.01).结论三种方法检测鼠疫F1抗原诊断鼠疫,RIHA结果有定量的意义,非特异性凝集率高,复试工作量大;GICA和ELISA具有相同的特异性和敏感性,GICA结果只有定性意义,ELISA排除了RIHA、GICA两种方法的缺陷,综合了两种方法的优点.     

中蒙边境阿尔泰山区鼠疫调查研究

目的 探索中蒙边境阿尔泰山区鼠疫自然疫源地，为做好边境地区疫情防控提供依据。方法 在中蒙边境阿尔泰山区建立野外帐篷实验室鼠疫监测点，用1日弓型夹法在自毙鼠洞、洞旁、周边洞群布局捕鼠，采用胶体金免疫层析(GICA)、上转发光(UPT-3A)、微量血凝(IHA)或(RIHA)、酶联免疫吸附(ELISA)4种方法，检测所获标本鼠疫菌F1抗原和抗体携带情况。结果 监测中共捕获小兽类5科7属7种163只。首次从中蒙边境阿尔泰山花海子区捕获的长尾黄鼠中检出鼠疫菌F1抗原阳性1份、抗体4份，在9只林鼬中检出鼠疫菌F1抗体阳性8份、在赤狐中检出F1抗体阳性2份。阳性检出点分为3片，总面积约0.357 hm²。为控制疫情扩散，启动了鼠疫预警应急预案，及时开展了处置。结论 首次发现中蒙边境阿尔泰山区存在动物鼠疫，应进一步加大调查研究，查清该区域鼠疫分布和流行规律，为当地做好鼠疫防控提供科学依据。     

鼠疫FI单克隆抗体杂交瘤株的建立

先后两次分别用鼠疫EV活菌和FI抗原免疫BALB/C小鼠的脾细胞与SP2/0瘤细胞混合,大容量40％PEG处理后离心法杂交,用IHA和PPA-ELISA检测McAb,建立了21株分泌抗鼠疫FI抗原的McAb的杂交瘤株,初步鉴定了其中5株。观测了1株杂交瘤分泌的McAb用于RIHA的特异性和敏感性。     

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.