BACKGROUND: The relation of epilepsy to immunity has been investigated at cellular and molecular levels in recent years, and the results show many immunological changes in epileptic patients such as immunocytes,immune molecules and immune functions.OBJECTIVE: To investigate the immunologic pathogenesis of epilepsy and expression of MHC- Ⅰ, MHC- Ⅱ and C3b receptor of microglia. DESIGN: Randomized and controlled trial. SETTING: Medical College of Dalian University; University of Sanchong,Japan.MATERIALS: The experiment was conducted in the Immunology Laboratory of Jilin Beihua University from 2001 to 2002. Forty adult Wistar rats were randomized into model group and control group with 20 in each group.METHODS: 12 mg/kg kainic acid was administered subcutaneously into rats in model group, while no intervention was given to rats in control group. Seizure was observed within 6 hours following kainic acid administration. Rats were killed 3 days after medication. Neuronal degeneration was observed with crystal violet staining and the expression of MHC molecules and C3b receptor in the hippocampus of epileptogenic rats was observed immunnhistochemically.administration, spasm occurred within 3 hours after kainic acid administration and continued from the first stage to the fifth stage. More severe served in cone-like cell layer of hippocampus of rats in model group, but molecule, MHC- Ⅱ molecule and C3b receptor was (201.6±6.43), (493.8±7.92) and (362.5±3.18) cells per visual field respectively in the hippocampus of epileptogenic rats inmodel group, but no obvious expression was observed in control group. The differences were significant between the two groups (P ＜ 0.01).pocampal sclerosis, immunological inflammation is observed combined with the complement system, which indicates immunologic inflammatory mechanism of epilepsy.

BACKGROUND: The recent researches show that there are various kinds of autoantibodies in epilepsy patients such as anti-nuclear antibodies, anticardiolipin antibodies,anti-?2 glucoprotein antibodies and anti-glutamic acid decarboxylase antibodies,which indicate that some types of epilepsy are mediated by autoimmune mechanism.OBJECTIVE: To detect the level of anti-glutamic acid receptor (GluR1)antibody in epilepsy children.DESIGN: A controlled trial.SETTING: Medical College of Dalian University, General Hospital of Jilin Chemistry Group and Sanchong University, Japan.PARTICIPANTS: Totally 56 cases of epilepsy children were from Maternal and Child Health in Qingdao City including 30 males and 26 females and aged from 3 months to 13 years.While 48 health examinees and 12 patients with brain tumor from General Hospital of Jilin Chemistry Group were taken as controls, including 38 males and 12 females and aged from 6months to 17 years.Their guardians were all informed of the detection index and consented to join the study.METHODS: Polystyrene plates were coated with GluR1 polypeptide antigen,and optimal conditions of enzyme linked immunosorbent assay (ELISA) for GluRI antibody was ascertained with chessboard titration.MAIN OUTCOME MEASURES: The GluR1 antibody level was detected with ELISA.RESULTS:The concentration of coating GluR1 polypeptide antigen was20 mg/L; The dilution of serum and horseradish peroxidase sheep anti-hu-man (HRP-SaH) IgG were 1:50 and 1:1 500 respectively. Intra-assay andinter-assay coefficientof variability were 4.48％-8.80％ and 11.18％-16.60％ respectively; The specific absorption test demonstrated that the ab-sorbance value of positive serum decreased 2.4 times after absorption ofGluR1 polypeptide compared with that before absorption.There was obvi-ous higher positive rate of GluR1 antibody in serum of epilepsy childrenthan control group (40％, 5％, P＜0.01).CONCLUSION:①ELISA is specific and stable for GluR1 antibody in serum. ②There is the possible autoimmune response of GluR1 antibody in serum of epilepsy children.

Objective:To apply immuno-PCR for detection of anti-Sm antibodies.Methods: Immuno-PCR was established for anti-Sm antibodies in SLE sera using SaHIgG-DNA probe. The experimental conditions were optomized and methological identifieation indicated obvious in crease(×107 ) in sensitivity compared with that of ELISA. The positive rate of anti-Sm antibodies was 54.9% in SLE sera, and significantly higher than that of ELISA. Intra-and inter-CV were 2.3%～4.8% and 3.7%～6.9% respectively. Immuno-PCR was positively correlated with ELISA( p ＜0.001). Therefore, immuno-PCR for anti-Sm antibodies is specific,sensitive,reproducible and practical clinically.

Based on the medical immunology exquisite course of Dalian city,we expound the guiding thought,construction and system evaluation of medical immunology test bank,in order to normalize examination,consummate teaching quality control system and improve teaching quality.

AIM: Although the evidence indicates that extracellular matrix metalloproteinase inducer (EMMPRIN) is closely associated with matrix metalloproteinase (MMP) expression in tumor cells, tumor invasion and metastasis, no direct proof that EMMPRIN regulates MMPs in monocytes, especially in the atherogenic milieu is observed. Here we tested this hypothesis by examining MMP-9 expression in macrophages/foam cells and monocyte migration through EMMPRIN knockdown by siRNA. METHODS: The methods of qPCR and Western blotting were used to detect the suppressions of EMMPRIN mRNA and protein expression in macrophages and foam cells transfected with EMMPRIN-specific siRNA. The protein expression of MMP-9 in macrophages and foam cells was also determined. Monocyte migration after EMMPRIN knockdown was observed by a Transwell assay. RESULTS: EMMPRIN knockdown by siRNA markedly abolished the MMP-9 expression by 50% and 40% in macrophages and foam cells, respectively. Migration induced by chemotactic factor MCP-1 and VEGF was significantly attenuated (P<0.05) in monocytes treated with EMMPRIN-siRNA. CONCLUSION: The protein expression and secretion of MMP-9 are down-regulated by EMMPRIN knockdown during monocyte differentiation into macrophages and foam cells. Moreover, EMMPRIN siRNA treatment also prevents monocyte migration. Thus, EMMPRIN plays a key regulatory role for MMP activity and monocyte migration, making it a potential target for pharmacological intervention of atherosclerosis.

Objective To study the possible influencing factors in the formation of coronary collateral circulation in patients with chronic total occlusion (CTO). Methods Patients were enrolled having at least 1 major coronary artery angiography revealed as CTO of 144 patients. According to the Rentrop classification, patients with grade 0 and grade 1 filling were catogorized as insufficient collateral circulation group (n=72) and patients with grade 2 and grade 3 filling as collateral circulation group (n=72). Serum biomarkers and insulin-resistance by HOMA model were also studied in all patients. Results In the insufficient collateral circulation, BMI,TC,ApoB, lipoprotein a, fasting insulin HOMA-IR,HOMA- beta, CRP was significantly higher than the well collateral circulation group and ApoA-Ⅰ, ISI lower than the well collateral group ( all P ﹤0. 05 ) . Bivariate correlation alaysis showed. Rentrop score, BMI, TC, ApoB, lipoprotein a, fasting insulin, HOMA-IR,HOMA- beta and CRP are positively correlated to the formation of collateral circulation ( P ﹤ 0. 05 ); ApoA-Ⅰ and ISI were negatively correlated ( P ﹤0. 05 ) . Logistic regression analysis after calibration with weight, ApoA-Ⅰ and HOMA-beta factors, lipoprotein a ( OR 7. 575,P=0. 009), TC (OR 2. 154,P =0. 001) were found to be the independent factors of coronary collateral circulation formation. Conclusions TC, lipoprotein a, obesity, CRP, and HOMA-IR are correlated with the formation of coronary collateral circulation and may predict formation of collateral circulation in patients with CTO.

Objective To explore the effects of smoking on serum paraoxonase1 (PON1) activity and carotid atherosclerosis.Methods 478 subjects from residents of health screening for cardiovascular disease were enrolled from June 2012 to July 2014 in Huangpu district,shanghai.Smoking,drinking,exercise and cardiovascular disease risk factor data were recorded and gathered.All subjects accepted carotid artery ultrasound examination and were measured serum PON1 activity.The lowest quartile of serum PON1 activity level was taken as low PON1 activity level.Results (1) Serum PON1 activity in smokers was lower than that in non-smokers ((206.5±25.6) kU/L vs (230.9±38.1)kU/L,P＜0.01),incidence of carotid artery atherosclerosis in smoking group was higher than non-smoking group(75.7％ vs 56.1％,P＜0.01).(2) Multivariate logistic regression analyses indicated smoking,lack of exercise,creatinine,LDL-C were the independence factors of PON1 activity.(3) Multivariate logistic regression analyses indicated smoking,serum PON1 activity,age,gender,systolic pressure,were the independence factors of carotid atherosclerosis.Conclusion Smoking reduces serum activity.Smoking and lower serum PON1 activity level are independent risk factors of carotid atherosclerosis.

AIM: To investigate the role of PPAR ? or ? ligands in regulating the expression of extracellular matrix metalloproteinase inducer(EMMPRIN).METHODS: THP-1 monocytes were induced into macrophages and foam cells in vitro then interfered with clofibrate and pioglitazone.The cells and supernatant were collected after 24 h,respectively.EMMPRIN gene and its protein were assessed by real-time PCR and Western blotting in different interferences.The concentration of matrix metalloproteinases(MMP-9) was measured with ELISA method and the activity of MMP-9 was detected with gelatin zymography.RESULTS: Two known PPAR ? or ? ligands,colfibrate and pioglitzaone,were found,both of which inhibited EMMPRIN expression in macrophages and foam cells.The inhibition was correspondent to the secretion and activity of MMP-9 simultaneously.CONCLUSION: Both PPAR ? and ? ligands inhibit EMMPRIN expression,which may account for their effect on inhibition of MMPs.

AIM: To determine the effect of intermittent hypoxia (IH) precondition on ischemic myocardium by using a rabbit model of chro nic myocardial ischemia with left anterior descending (LAD) banding. METHODS: Male, adult New Zealand white rabbits were assigned into three groups randomly. (1) normal group (N group), (2) control group (C group), (3) IH precondition group (H group). A LAD band was placed in C and H group firs t. The rabbits in H group were exposed to altitude of 5 000 m, for 6 h/day c ontin uously. According to IH precondition duration, the animals were subdivided into 7-days group (H1) and 42-days group (H2). After these experiments, the mRNA conc entrations of vascular endothelial growth factor (VEGF), hypoxia induced factor (HIF-1?), endothelial nitric oxide synthase (eNOS) and the expression of VEGF p rotein were detected. Tissue sections were stained for alkaline phosphatase with indoxyl-tetrazolium method to detect capillary density. RESULTS: The mRNA levels of VEGF, HIF-1?, eNOS and expression V E GF protein were increased in H group significantly. Compared with C and N group, the capillary density in H group was increased significantly. CONCLUSION: IH precondition increases angiogenesis in chronic is chemic myocardium in rabbits.

Objective To investigate the relationship between extracellular matrix metalloproteinase inducer (EMMPRIN), the upstream regulatory factor of matrix metalloproteinase (MMPs), and the formation of atherosclerosis and the clinical type of coronary heart disease. Methods A total of 223 patients were classified into four groups according to results of coronary angiography (CAG) and clinical data: STEMI group (65 patients with ST-segment elevation myocardial infarction), NSTE ACS group (42 patients with non-ST-segment elevation acute coronary syndrome), SAP group (75 patients with stable angina pectoris) and normal control group (41 patients of CAG-negative). The mean fluorescence intensity (MFI) of EMMPRIN on monocytes of peripheral blood (PBMCs)were examined by flow cytometry. MMP-9 in serum was measured with ELISA; high-sensitivity C-reactive protein (hs-CRP) in serum was measured with immune velocity method. Results The EMMPRIN MFI on PBMCs in SAP group, STEMI group and NSTE ACS group was higher than that in the normal control group (P<0.05 or P<0.01). The EMMPRIN MFI in STEMI group and NSTE ACS group was higher than that in SAP group (P<0.05 or P<0.01). The expression characteristic of EMMPRIN on the PBMCs was consistent with that of hs-CRP and MMP-9 in each group. The EMMPRIN MFI of the PBMCs had positive correlation with the level of MMP-9 and hs-CRP in serum (r=0.168,P<0.05;r=0.305,P<0.01). Conclusion EMMPRIN may has promotive effect on the formation of atherosclerosis and unstablility of coronary heart disease as an upstream regulatory factor of MMPs