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C.jejuni分离及周围神经不同状态病理变化

The Method for Isolating Campylobacter Jejuni and Peripheral Nerve’s Pathology of Different State

【作者】 付金生

【导师】 李震中;

【作者基本信息】 河北医科大学 , 神经病学, 2012, 硕士

【摘要】 第一部分GBS及非腹泻住院患者空肠弯曲菌携带情况研究目的:1、了解临床诊断的吉兰-巴雷综合征(Guillain-Barre syndrome,GBS)患者便标本空肠弯曲菌(Campylobacter jejuni, C.jejuni)携带情况;2、了解非腹泻住院患者便标本空肠弯曲菌携带情况。方法:1、标本采集:无菌棉签采集河北医科大学第二医院心血管内科、神经内科、消化内科、小儿内科、血液内科、内分泌科、皮肤性病科、腺体外科、胃肠外科、干部病房、呼吸内科11个科室的非腹泻住院患者的便标本,包括GBS患者,量约黄豆粒大小,放入盛有8mL布氏肉汤的西林瓶中,立即带回实验室;2、标本培养及细菌分离:将带回实验室的西林瓶立即置于42℃、微需氧环境(5%O2、10%CO2、85%N2)增菌培养,以24h为一周期,进一步应用分离平板传代,分离疑似空肠弯曲菌;3、鉴定方法:对分离得到的疑似空肠弯曲菌菌株,通过形态学、生化反应、分子鉴定等方法进行鉴定,确定为空肠弯曲菌后增菌保存;4、统计方法:统计标本来源的病例数和分离得到的阳性菌株数,对GBS患者及非腹泻住院患者分别统计便标本空肠弯曲菌携带阳性率。结果:1、8例临床诊断的吉兰巴-雷综合征患者便标本培养、分离空肠弯曲菌,通过鉴定得到1株目的菌株;2、274例非腹泻住院患者便标本培养、分离,未见阳性空肠弯曲菌分出。结论:1、本实验对河北医科大学第二医院收住的临床诊断为GBS的8名患者的便标本进行空肠弯曲菌的培养分离,得到1株目的菌株,但对其与吉兰-巴雷综合症之间的关系及其致病性有待进一步研究;2、其他非腹泻住院患者的便标本未见可疑空肠弯曲菌分出,可能由于细菌状态、技术方法等方面的限制,出现假阴性结果,故本方法不合适该人群的空肠弯曲菌分离。第二部分周围神经沃勒变性病理观察及死亡后病理变化目的:1、认识周围神经沃勒变性(Wallerian degeneration)病理变化过程;2、鉴别死亡后周围神经病理变化,以区别动物模型疾病本身导致的病变和死亡后随时间延长自然病理变化。方法:1、建立沃勒变性动物模型,分别于神经切断后1h、2h、4h、6h、8h、12h、24h、2d、4d、6h、8d、15d行坐骨神经取材,远端神经同部位进行饿酸染色,光镜下观察病理变化;2、分别于小鼠活体及处死后1h、2h、4h、6h、8h、12h、24h、48h取材坐骨神经,行饿酸染色,观察病理变化。结果:1、神经切断后1h,远端神经可出现髓鞘不光滑,随时间延长,髓鞘逐渐回缩,2d时卵圆体形成,出现典型病变;2、周围神经随死亡时间的病理变化:小鼠处死后4h,坐骨神经行饿酸染色病理所见基本正常,6h出现病理性人为损伤,如牵拉损伤,随时间延长人为损伤增多,逐渐出现不规则破坏、虫蚀样等改变。结论:探讨周围神经沃勒变性和随死亡时间的病理变化,应用饿酸染色技术可动态观察其变化过程,可区分二者的典型病变。

【Abstract】 Part1The study of carrying rate toward GBS and non-diarrhea ofhospitalized patients with Campylobacter bacteriaObjective:1To study the carrying of Campylobacter jejuni (Cj) in thestools of the clinical Guillain-Barre syndrome (GBS).2To study the carryingof Cj in the stools of hospitalized patients which have no diarrhea.Methods:1The collection of the specimen: with sterile cotton swab,collect the specimens of the hospitalized patients in the Department of Cardio-logy, Neurology, Respiratory, Gastroenterology, Pediatrics, Hematology, End-ocrinology, Dermatology and the surgery of gland, Gastrointestinal, Cadreward of the Second Hospital of Hebei Medical University, including thepatients of GBS. The volume of the collection is about a bean. The specimenswere placed into penicillin bottles which contain8milliliters of Brucella brothand brought back to laboratory immediately.2The culture and isolate of thespecimens: the penicillin bottles were cultured in a micro-aerophilic atmosph-ere (5%O2、10%CO2、85%N2) at a temperature of42℃With a cycle of24h,to isolate the suspected Cj with a selective medium.3The methods ofidentification: by the methods of morphology, biochemical reactions andmolecular, the suspected Cj isolated were identified and then saved.4Statistical methods: counted the number of the cases and positive strains.Calculated the positive rate of GBS and hospitalized patients which have nodiarrhea.Results:1Obtained one suspended Cj from the stools of8clinical GBSand the identification results is positive.2there is no Cj isolated from thestoos of274hospitalized patients which have no diarrhea.Conclusions:1In this study, we admitted8clinal GBS and isolated1 strain of Cj from one of them. But the association between the strain and GBSneeds to be further studied.2There is no Cj isolated from the stools of otherhospitalized patients which have no diarrhea. The false negative results maybe due to the state of the bacteria, technical methods, so the methods of isolat-ion is inappropriate for this populations. Part2The pathological result of Wallerian degeneration and afterdeath in Peripheral nerveObjective:1Recognize the pathological result of Wallerian degenerate-on as for Peripheral nerve;2Recognize the pathological changes of the perip-heral nerve after deaths. Identify the differences between pathological changesby animal model and the natural pathological changes with time after the dea-ths.Methods:1To establish the animal model of wallerian degeneration andtake the sciatic nerve at the times of1h,2h,4h,6h,8h,12h,24h,2d,4d,6h,8d,15d after nerve transection. Then evaluate the pathology through osmicacid.Results:1After1hour of nerve transection, The myelin of distal nervebecame not smooth. Along with the time, the myelin gradually retracted. when2days after nerve transection, oval body appears;2The pathological changesof peripheral nerve after deaths: when4hours after deaths, pathology throughosmic acid is almost normal. when6hours, artificial destruction appears, suchas traction. Along with the time, irregular destruction appears gradually, suchas worm-eaten-like.Conclusions: Recognize pathology of Wallerian degeneration and pat-hological changes of the peripheral nerve after deaths. Dynamic changes canbe observed through osmic acid.

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