Aim In order to improver the diagnostic rate of Prion disease, solve the lacking sources of nature PrP antigen. ,synthesize the human PrP peptide, prepare the antibody to PrP peptide further. Method A synthetic peptide with the sequence identical to the 15 residues of human PrP, as described by Kretzschmar[1], was synthesized by the solid-phase method. The synthetic peptide was coupled to bovine serum albumin(BSA)by the method of EDCI〔2〕. The polypeptide combined with BSA was used as antigen to immunize the rabbit and detected by immunomethod. Result The PrP polypeptide combined with BSA obtained immunogenicity and anti-PrP synthetic peptide antiserum was successfully obtained. Conclusion The preparation and application of human PrP synthetic peptide can substitute for nature PrP antigen partly. It has laid a foundation for further preparation of monoclonal antibody to PrP and the study of Prion disease.

BACKGROUND: The increase of concentration of plasma homocysteine is the independent risk factor of atherosclerotic and thrombotic cerebral infarction. Genic mutation of methylene tetrahydrofolate reductase (MTHFR), which is the metabolic enzyme of homocysteine in thansulfuration and remethylation,can induce the elevation of the concentration of plasma homocysteine.OBJECTIVE: To explore the relationship of hyperhomocysteinemia and genic mutation of MTHFR of homocysteine with ischemic cerebrovascular disease in youths.DESIGN: Case-control observation,SETTING: Department of Neurology, China-Japan Friendship Hospital,Jilin UniversityPARTICIPANTS: 100 young patients with cerebral infarction, who were hospitalized within 2 days after episode at the Department of Neurology,China-Japan Union Hospital, Jilin University between April 2003 and December 2004, were enrolled as case group, 73 males and 27 females, aged 27-45 years old with an average of (42±5) years. 100 cases in control group were healthy people receiving health examination in the same period, 70 males and 30 females, aged 18-45 years old with an average of (39±4) years.METHODS: The homocysteine in fasting plasma of testees was detected with high performance liquid chromatograpy (HPLC). C677T site and A1298C site of MTHFR gene were analyzed with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and detected with armplification refractory mutation system (ARMS).MAIN OUTCOME MEASURES: Detection of MTHFR C677T and A1298C gene; Relationship between concentration of plasma homocysteine and MTHFR genotype.RESULTS: A total of 100 inclusive patients and 100 normal control people were involved in the result analysis. ①Detection of MTHFR C677Tand A1298C gene: Distribution of genotype, frequency of homozygote and frequency of allele of MTHFR C677T in the case group and control group had significant difference (P ＜ 0.01 ) while the distribution of genotype,frequency of homozygote and frequency of allele of MTHFR A1298C gene in the case group and control group had insignificant difference (P ＞ 0.05 ). ②Relationship between the concentration of plasma homocysteine and MTHFR genotype: The concentration of plasma homocysteine between MTHFR C677T and A1298C genotype had significant difference (P＜ 0.001 ). The mutant result LSD-t check of the 2 sites showed that mean difference of homozygote and heterozygote, homozygote and concentration of wild type homocysteine had statistical significance (P ＜ 0.05). The mean difference of MTHFR C677T and A1298C heterozygote and concentration of wild type homocysteine in plasma had no statistical significance (P＞ 0.05 ).CONCLUSION: The mutation of MTHFR C677T and A1298C leads to the marked increase in the concentration of plasma homocysteine. The MTHFR C677T polymorphism site is the independent risk factor of is chemic cerebrovascular disease in youths. The genic mutation of MTHFR A 1298C has no correlation with the attack of ischemic cerebrovascular disease of youths.

Objective:To study the Cronassial′protective effect on brain tissue during ischemia reperfusion.Methods:The rat cerebral ischemia reperfusion model was prepared by four-vessel occulasion method.The Glu,GABA content in hippocampus during cerebral ischemia reperfusion was observed by high press liquid phase method.Results:Glu,GABA content increased during ischemia and decreased during reperfusion. Cronassial could obviously decrease the Glu content and increase neuron density.Conclusion:Cronassial can inhibit Glu and protect brain tissue during cerebral ischemia reperfusion.

BACKGROUND: Cell apoptosis and expression of related apoptotic gene are found in rats with acute cerebral ischemia-reperfusion injury.OBJECTIVE: To study the effects of ganglioside on cell apoptosis in rats with acute cerebral ischemia-reperfusion injury.DESIGN: Randomized control animal experiment.SETTING: Department of Neurology, China-Japan Friendship Hospital of Jilin University.MATERIALS: The experiment was conducted in the Animal Laboratory of China-Japan Friendship Hospital of Jilin University at April 2002.Forty-eight healthy male Wistar rats aged 3-4 months with the body mass of (220±50) g were selected and randomly divided into ischemia-reperfusion group and ischemia-reperfusion + administration group (intraperitoneal injection of ganglioside GM-1 at 30 minutes before ischemia) with 24 rats in each group, and each group was subdivided into three groups according to the reperfusion time: 3-hour, 6-hour and 24-hour with 8 rats in each time-point.METHODS: ①Rat models of cerebral ischemia-reperfusion were established. ②Diphenylamine method was adopted to detect changes of DNA splitting rate in brain tissues at 3 hours,6 hours and 24 hours after cerebral ischemia.100 mg of cerebral cortex was made into 10% homogenate by adding into 0.9 mL of splitting fluid,which was then put in the centrifuge tube for repeated freezing and melting. The supernatant and deposit were collected.DNA splitting rate = supernatant absorption/(supernatant absorption + deposit absorption). ③Immunohistochemical method was used to study the expression of protein kinase Cδ (PKCδ) as well as changes of ganglioside after administration. MAIN OUTCOME MEASURES: Changes of DNA splitting rate in cerebral cortex of rats as well as intensity of PKCδ expression.RESULTS: One rat in the normal saline control group died for exceeding anesthetization at the 6th hour of reperfusion, and 2 rats died at the 24th hour of reperfusion, which were supplemented respectively. With the time of reperfusion increasing, changes of DNA splitting rate significantly increased, which peaked at the 24th hour. The expression of PKCδ peaked at the 6th hour of reperfusion and gradually decreased. The DNA splitting rate and PKCδ expression were remarkably decreased at corresponding time-points in the ganglioside group.CONCLUSION: Ganglioside can inhibit cell apoptosis and reduce PKCδ expression after cerebral ischemia-reperfusion to protect cerebral ischemia-reperfusion injury.

The changes ot free radicals matebolism in liver of rat fed grains from a Keshan disease endemic area and the effects of selenium and vitamin E on those were studied. The endemic grains that were deficient in selenium caused markedly decrease in the glutathione peroxidase activity, and marked increase in the lipid peroxides content and the free radicals level of the rat livers. The diets that supplemented the endemic grains with either selenium (0.22 mg/kg diets) or DL-alpha-tocopherol (100 mg /kg diets) caused the falls in the lipid peroxides content and the free rdicaals level. There was a marked rise in the al pha-tocopherol content and reduced glutathione content in livers of rats fed grains from the endemic area, suggesting an increased reqiu-rement of vitamin E and production of glutathione in the selenium-defi-cient rats. The results indicate that a disorder of free radicals metaboli- sm is induced by the pathogenic factors existing in the grains cultiva-ted in a Keshan disease endemic area, and is reduced by supplementing the grains with sodium selenium or vitamin E, suggesting that relative lack of vitamin E may play an important role in the mechanism of Keshan disease onset.

Objective To observe protective effects of allicin on heart and lung in aging mice induced by D-galactose, and try to explore the possible mechanism from the aspect of oxidative stress. Methods Fifty male Kunming mice were randomly divided into 5 groups:normal control group ( group C) ,model control group ( group D) and high-,middle-and low-dose of allicin groups ( H,M,L group) . The rats in group D,L,M and H were injected with D-galactose. At the same time,the rats in group L,M and H were injected with allicin. All of the injection lasted for 6 weeks. After that,athletic ability of the mice was evaluated by behavioral experiments. Malondialdehyde ( MDA ) content, superoxide dismutase ( SOD ) activity and total antioxidant capacity ( T-AOC) in the homogenate of heart and lung tissues were measured,and the expression levels of Bax and Bcl-2 were tested in heart and lung. Results Behavioral experiment showed that standing time on Rota Rod System and continuous running time on mice treadmill were significantly shortened in group D as compared with group C ( P<0.05) ,but those were prolonged in the allicin groups. Compared with group C,MDA content in homogenate of myocardium and lung tissues was significantly increased in group D (P<0.05),the activity of SOD was decreased (P<0.05) and T-AOC was decreased (P<0.05) . As compared with group D,MDA content in allicin groups was significantly decreased ( P<0.05) ,SOD activity increased (P<0.05) and A-TOC increased (P<0.05) in a dose-dependent manner. The results of immunohistochemistry showed that the expression of Bax was significantly increased in group D as compared with group C,and the expression of Bcl-2 was decreased. After the intervention of allicin,expression of Bax was decreased in group L,M and H as compared with group D, and expression of Bcl-2 was increased as compared with group D. Expression levels of Bax and Bcl-2 in lung tissues showed the same changing trend. Conclusion Allicin has a potential protect role on heart and lung in D-galactose-induced aging mice by increasing athletic ability,decreasing oxidative stress and decreasing cell apoptosis of myocardium and lung tissues.

Objective To summarize the sonographic features of fetal limb deformity. Methods Systematic continuous sequence approach (SCSA) was performed with two-dimensional and three-dimensional ultrasonography(USG) in 28 383 fetuses to observe the fetal limb development, posture abnormality and other accompanied malformations. Compared with the pathological and radiological findings, the characteristics of fetal limb deformity on USG were summarized. Results Among 28 383 fetuses prenatal ultrasound detected 207 cases of fetal malformations (0.7%, 207/28 383) including 29 cases of limb deformities (14%, 29/207). In the 29 cases, there were osteogenesis imperfecta in 2 cases, syndactyly in 1 case, cleft hand deformities in 1 case, uncifom hand in 1 case, clubfoot deformity in 12 cases, cleft foot in 1 case, micromelia in 4 cases, limb body wall complex in 1 case, forearm defect in 2 cases, and radius absence in 4 cases. Chromosome karyotype analysis was conducted in 7/29 cases, of which 6 cases were normal and 1 case was trisomy-13 with syndactyly. In addition, the fetal limb deformities were found at 17-19 weeks of gestation in 4 cases, at 20-24 weeks in 23 cases, and at 25-33 weeks in 2 cases. In summary, 27/29 cases were identiifed at 17-24 weeks of gestation. Conclusions Prenatal ultrasound is the ifrst-choice method for screening of fetal limb deformity. The detection rate of limb deformity could be greatly improved by using SCSA method with the supplement of 3D ultrasound.

Objective:To observe the effects of myricetin on autophagy and mitochondrial fission in the human ovarian cancer SKOV3 cells,and to explore its induction effect on autophagy and promoting effect on mitochondrial fission.Methods: The SKOV3 cells were cultured in vitro.0,20,40,and 60 g·L-1 myricetin were used in control group and low, middle, high doses of myricetin groups for 12 h.The changes of mitochondrial membrane potential were detected by flow cytometry;the morphology of mitochondria was observed by MitoTracker Red;the expression levels of dynamin related protein1 (Drp1) and fission 1 (Fis1) in various groups were detected by Western blotting method;and the expression levels of autophagy related protein LC3 were detected by both Western blotting method and immunofluorescence method.Results:Compared with control group, the ratios of decreased mitochondria in different doses of myricetin groups were significantly increased(t=3.27, t=6.85, t=5.49,P<0.05).Compared with control group, the numbers of mitochondria in different doses of myricetin groups were increased, and the mitochondria looked more like gravel.Compared with control group,the expression levels of Drp1 in different doses of myricetin groups were significantly increased (t=4.35, t=3.28, t=6.17,P<0.05), and the expression levels of Fis1 were increased also(t=8.32, t=6.74, t=9.27).The immunofluorescence results showed that the expression levels of LC3 in different doses of myricetin groups were significantly increased with the increase of myricetin dose compared with control group.The Western blotting results showed that the ratios of LC3-Ⅱ/LC3-Ⅰ in middle and high doses of myricetin groups were significantly increased compared with control group(t=3.28, t=4.21,P<0.05).Conclusion: Myricetin can induce the autophagy of SKOV3 cells, and it can also promote the mitochondrial fission.

ObjectiveToinvestigatetheeffectandmechanismofaChinesetraditionalprescription,YuebiBanxia decoction, on allergic asthma in mice by observing the changes of serum immunoglobulin E ( IgE ) and lung tissue superoxide dismutase ( SOD) activity in the mouse model of allergic asthma .Methods Forty healthy Kunming mice were randomly divided into normal control group , asthma model group , dexamethasone intervention group and Chinese medicine preparation group .The mouse model of asthma was generated by peritoneal injection of ovalbumin ( OVA) and suspension of aluminium hydroxide .The mice of traditional Chinese medicine group received Yuebi Banxia decoction in a dose of 1 mL/100 g, daily for 30 days.The mice of dexamethasone group were given intraperitoneal injection of dexamethasone 1 mg/kg.The normal control group and asthma model group were given gastric gavage of physiological saline.After 30 days administration, the serum levels of IgE and SOD activity of lung tissue were determined , and the pathological changes of lung tissue were observed using HE staining .Results The mice of traditional Chinese medicine preparation group showed significantly decreased serum IgE level , and significantly increased SOD activity than those of the asthma model group mice (P<0.05 for both).Conclusions Reduced serum IgE and increased SOD activity in the lung tissue of mice with allergic asthma , improving the antioxidant ability of lung tissue , may be involved in the anti-allergic asthma mechanism of the Chinese traditional prescription Yuebi Banxia decoction .

Objective To study the effect of infrasound on expression of calmodulin-dependent protein kinase II (CaMKII) and tau pro-tein in hippocampus of rats. Methods Fifty-six male Sprague-Dawley rats were randomized into control group (n=8), 1-day group (n=8), 7-day group (n=8) and 14-day group (n=32), and the 14-day group was subgrouped as 1-hour, 6-hour, 24-hour, 48-hour subgroups, naming after the time after infrasound exposure, 8 in each subgroup. All the test groups were put in an infrasound field with 8 Hz, 130 dB for 2 hours daily, while the control group was put in the infrasound instrument without infrasound exposure for 2 hours daily. The expression of pT286-CaMKII and tau protein in hippocampus was detected with immunohistochemisty, Western blotting and enzyme-linked immunoabsor-bent assay. Results The expression of pT286-CaMKII was the most in 14-day group (F>14.912, P<0.001), as well as the expression of tau pro-tein (F>36.229, P<0.001), and secondary in 7-day group (P<0.05). For 14-day group, the expression of tau protein was the most in 1-hour and 6-hour subgroups, and dropped down in 24-hour subgroup, although more than that in the control group (P<0.05). Conclusion Exposure of 8 Hz, 130 dB infrasound may induce phosphorylation of CaMKII and tau protein, and the expression of tau protein in hippocampal cells in rat, which may disturb their learning and memory function.