| Objective To establish a H.pylori infected BALB/C mouse model ;To assess the effects and the mechanism which immunization were combined with traditional Chinese against Helicobacter pylon infection. Methods (1)H.pvlori were cultured and identified Sydney strain 1 H.pylori (881), presented by Sun Yat -Sat University of medical Sciences, was cultured on medium ,and incubated under micro-aerobic condition (O2 5%, CO2 10%, N2 85%) at 37℃ for 3-5 days; H.pylori was identified by germ figure ,rapid urease test and histology (2) The establishment of mouse model: Forty-three Specific Germ Free (SPE) BALB/C female mice (7-8 week,17-20 grams) were randomly divided into two groups. Thirty mice were served as infected groups. thirteen mice were served as control group. Mice in the experimental group were5inoculated with SS1 H.pylori, 0.4ml of per mouse, five times for ten day. Mice in the control group were not received any inoculation. In 4, 8,16,22 weeks after the last inoculation, seven animals in the experimental group and three animals in control group were sacrificed respectively. Biopsies were taken for urease test, histology studies to assess the H.pylori colonization and the change of the pathology. (3)The experiment of therapeutic immunization and traditional Chinese :70SPF BALE/C female mice were inoculated the SS1 H.pylori, O.4ml of inoculated per mouse ,four times in ten days In 8 weeks after the last inoculation,5animals were randomly sacrificed, they were all approved to have been infected by H.pylori by urease test and histology studies .Then , mice were randomly divided into 6 groups ,A group had 13 mice and were treated with urease plus GTE and HAP, B group had 13 mice and were treated with urease plus CTB HAP and traditional Chinese ,C group had 13 mice and were treated with traditional Chinese alone ,D group had 13 mice and were treated with urease alone , E group had 6 mice and were treated with GTE HAP ,F group was control group and had 6 mice . Therapeutic dosage: urease 250ug CTB 4ug HAPImg; Wei Tai were executed into liquor (concentration: O.128g/ml). Urease GTE HAP oral once a week for four weeks, Traditional Chinese oral once a day for two weeks .(4)Test figure: Four weeks after the last treatment , all animals were sacrificed. Peripheral 2-2.5m1 blood were collected into two cuvettes, the cuvettes which had heparin were tested I lymphocyte by FACSCaIibur FCM,the6cuvettes no heparin were taken to collect serum and to test anti-H.pylori-IgA; Stomach biopsies were collected to detect H.pylori eradicaton and the change of the pathology by urease test and HE Giemsa stain. RESULT:(I) The establishment of H.pylori infected mouse model: H.pylori were detected in all mice in the experiment group by the urease test and histology in week 4 ,8 ,16,22 .AJI animals were H.pylori negative in control group respectively .A few of 1-l.pylori colonization were detected in stomach mucosa in 4,8 weeks ,The number of H.pylori colonization tended to increase over the experiment period . All animals had no inflammatory changes in week 4,8.However in week 16, a few of lymphocyte in stomach mucosa. In week 22, mild to moderate chronic active gastritis was observed,Many lymphocyteeosinophuls and a few of neutrophils were detected in submucosa of stomach. On the contrary,in the control group,no remarkable changes were detected(2)The experiment of therapeutic immunization and Wei Tai:OEradication rate of H.pylori :A group was 61.54,B group was 75,C group was53.85,the other three groups were zero.(2)The changes of pathology :few of inflammatory cells were observed in suhmucosa of stomach in A, 13, C groups In other three groups many lymphocyte, eosinophi.ls and a few of neutrophils were detected obviously. (3)T lymphocyte subsets in peripheral blood: CD3, CD4 CD4/CD8 in immunization plus traditional Chinese group were significantly higher7than other groups CD3, CD4, CD4/CD8 in immunization and traditional group were higher than urease and control gro... |
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