Investigations Of Peripheral Nerve Lesions Caused By C.jejuni And Its Epidemiology

Part Ⅰ: Peripheral nerve lesions caused by C.jejuniObjective:Preliminary studies of the influence of different Immunization, animalspecies,bacterial species, different bacterial concentrations over the peripheralneuropathy in animal models of Guillain-Barre Syndrome (Guillain-Barresyndrome, GBS) caused by Campylobacter jejuni (Campyloba-cter jejuni, CJ).Methods:1Preparation of Bacterial suspension: isolate and culture theCampylobacter jejuni strains from the Guillain-Barre syndrome(Guillain-Barre syndrome, GBS) patients of North China. Take them fromthe refrigerator in which the temperature is-80℃and add them to the broth inthe condition of42℃microaerophilic (10%CO2,5%O2,85%N2), thenculture the stains for24~48h to recovery the bacteria. Passage the strains inColumbia blood plate which contains7%sterile and fiber blood. Scrape thecolonies after purification, mix it into Brucella broth or saline to get thebacterial suspension.2Experimental groups:GroupA: Immunize Balb/c mice by intraperitoneal injection, gastricperfusion, subcutaneous injection.Group B: Divide Balb/c mice into six groups and receive respectively thegastric perfusion of NCTC1116810×4cfu per mouse,10×6cfu per mouse,10×8cfu per mouse and Lulei10×4cfu per mouse,10×6cfu per mouse,10×8cfuper mouse.Group C: The same way to immunize Balb/c mice and KM mice. 3Stool culturesConsecutive stool cultures of Campylobacter jejuni taken from the micewho received no treatment,who received the gastric perfusion and whoreceived the multi-point intraperitoneal injection. Culture Campylobacterjejuni from the stools continuously from the2nd day of the treatment.4Neuropathological examination: use osmium tetroxide staining,observethe neuropathy. And analyze statistically the results.Results:1Symptoms and signs1.1We can see the animal who received the gastric perfusion has Diarrhea,the fur stands, eat and drink less, and their activities reduce.1.2The animal who received Intraperitoneal injection eat and drink less.1.3The animal who received intraperitoneal injection are listlessness andtheir activities reduced, the fur is messy and stands, The injection site is Redand swollen with ulcerate.No significant obstacles of physical activity were found in each group.2. peripheral neuropathyOsmium tetroxide acid stainA:The sciatic nerve,brachial plexus nerve of the three groups of miceshow neural shrinkage, myelin collapse.the percentage of single nerve fiberlesions of Subcutaneous group is41.5%,of oral group is10%,ofintraperitoneal injection group is27%.B: The sciatic nerve,brachial plexus nerve of groups of NCTC1116810x4cfu,10x6cfu,10x8cfu are smooth,there are no significant lesions.The percent-tage of single nerve fiber lesions of Lulei10x4cfu group is19%, of10x6cfugroup is30%, of10x8cfu group is30.5%.C:After immunize KM mice and Balb/c mice with the same immuniza-tion.Balb/c mice show obvious neuropathy, the incidence rate is34.3%. Theneural of KM mice is smooth and no neuropathy.3Stool cultures3.1No Campylobacter jejuni was seen in the group that received no treatment.3.2After the first gastric perfusion of live bacteria,Campylobacter jejuniin the stool cultures can be seen, the duration of CJ positive is2-10days; afterseveral times of gastric perfusion of live bacteria (more than3times,approximately2weeks per time) Campylobacter jejuni in the stool culturescan be seen, the duration of CJ positive is6-8days.3.3No Campylobacter jejuni was found in the subcutaneous and foot padmulti point injection group.Conclusion:1The lesions to Peripheral nerve in the Intraperitoneal injection group ismore serious than gastric perfusion group and the peritoneal injection. Thelesions of the peritoneal injection group is more serious than the gastricperfusion group.2Lulei can lead to peripheral nerve lesions,11168doesn't cause peripheralnerve lesions; the animal model of Lulei10x6concentration group is betterthan10x4concentration group, Lulei10x8group and Lulei10x6concentration arethe same on the nerve lesions to the mice.3Peripheral nerve damage caused by Campylobacter jejuni in Balb/cmice is more serious than that in KM mice.4The Campylobacter jejuni remained in the mice after the first gastricperfusion are greatly different,and of short duration, after several gastricperfusions, the rate of invasion success of Campylobacter jejuni increase andintestinal colonization time showed a steady growth trend. Part Ⅱ: Epidemiological investigations of C.jejuni innewborns and childrenObjective: To investigate the carrying of Campylobacter jejuni inthe stool of newborn and child.Methods:1Subjects:Choose healthy newborn that aged1d-9d in maternity ward andin patients in pediatric ward of the Second Hospital of Hebei MedicalUniversity from October to December.2Stool collection2.1Stool collection of newborns.After newborns defecation, picke a beansize stool with sterile cotton swab then put it into sterilized vials with boltonbroth,send it to laboratory in3hours.2.2Stool collection of pediatric patients.Ask patients to remain stool inSampling box, send it to laboratory in bolton broth in3hours.3Stool cultures:Separate and identify the Campylobacter jejuni from stoolreferring to《the laboratory standards of Clinical Isolation,identification andPreservation of Campylobacter jejuni》of the Second Hospital of HebeiMedical University Department of Neurology LaboratoryResults:Separat142cases of newborns stool, and40cases of children's stool.there are many Escherichia coli, Lactobacillus, but no Campylobacter jejuni.Conclusion:1The faeces of healthy newborns and children separated this time mayhave no Campylobacter jejuni.2The experimental methods we used may not applies to the separation ofCampylobacter jejuni of such populations.