Objective To investigate the location of CYLD in the neurons and explore the expression of CYLD in OGD/reperfusion-induced neuronal necroptosis.Methods Primary cortical neurons were cultured for 6days and neuronal purity was observed by double staining immunofluorescence of β3-tubulin and DAPI.The location of CYLD was identified by double staining immunofluorescence of NeuN,DAPI and CYLD using primary cortical neurons cultured for 14 days.Then,primary cortical neurons were divided into 8 groups:Control,EBSS,DMSO,OGD/reperfusion(0 h,2 h,6 h,8 h,12 h).Neurons were pretreated with zVAD-fmk for 30 min,OGD for 2 h and the levels of CYLD were evaluated after reoxygenation at different time points.The peak value(8 h) was chosen as reoxygenation time point.Neurons were divided into two groups as Control and OGD.The levels of CYLD were determined in both cytoplasm and nucleus after OGD 2 h and reoxygenation 8 h.Results The double staining immunofluorescence showed that neuronal cultured purity was about 70％ and the CYLD strongly expressed in nucleus but weakly in cytoplasm.The levels of CYLD increased gradually with different reoxygenation time and arrived at peak value after reoxygenation for 8 h (P ＜ 0.05),which was in accordance with the change of LDH (P ＜0.05) (Control (1.00±0.00),EBSS (1.07 ±0.03),DMSO (1.09 ±0.03),0h (1.40±0.12),2 h (1.74±0.08),6 h (2.25 ± 0.12),8 h (2.97 ± 0.15),12 h (3.01 ± 0.08)).The level of cytoplasm CYLD increased significantly in the OGD group (reoxygenation for 8 h)than that in control group (P＜0.05).But the level of nucleus CYLD had no difference between OGD and control group (P ＞ 0.05),which was in accordance with the results of immunofluorescence.Conclusion The CYLD in neurons cytoplasm is involved in necroptosis induced by OGD/deprivation and downregulating of CYLD has a protective effect on the brain injury resulted from ischemia/ reperfusion.

Objective To explore the value of Montreal Cognitive Assessment (MoCA) in the diagnosis of mild cognitive impairment. Methods Mini-mental state examination (MMSE) was used in 532 elderly persons aged 60 years and over in Xuzhou city Gulou county. The 69 healthy people and patients with mild cognitive impairment were chosen to undergo MoCA. Then the sensitivity and specificity of MoCA were analyzed. Results According to MMSE, there were 19 patients with mild cognitive impairment (27. 5 %), 50 healthy persons (72.5 %). While according to MoCA, there were 58 patients with mild cognitive impairment (84.1%), 11 healthy persons (15.9 %). The consistency of the two scales was not good. And compared with MMSE, the sensitivity of MoCA was 94.7%, and the specificity was 20. 1%. Conclusions In the diagnosis of mild cognitive impairment, MoCA is more sensitive than MMSE.

BACKGROUND:Psychiatric patients are the high risk group of people who are liable to commit suicide.Recently,research on neurobiology and genetics of suicide mainly concentrates in the study of 5 hydroxytryptamine(5 HT) system.It is found that T102C polymorphism of 5 HT2A receptor gene is associated with suicide in depressive patients. OBJECTIVE:To investigate the correlation of A 1438G polymorphism of 5 HT2A receptor gene with attempted suicide in psychiatric patients. DESIGN:An observational comparative study of taking the patients as subjects and healthy physical examinee as the controls. SETTINGS:A psychiatric department in a municipal hospital and a neurological department in a university hospital. PARTICIPANTS:All the samples in the research were collected from the Department of Psychiatry of Yangzhou Wutaishan Hospital from March to September 2002.Analysis of genotype was completed in the Neurobiological Laboratory of Xuzhou Medical Institute from October to December 2002.Inclusive criterion:patients met the diagnostic criteria of schizophrenia or affective disorder in the third edition of the Chinese Classification and diagnostic criteria of Mental Disorders(CCMD 3) and Diagnostic and Statistical Manual of Mental Disorders Fourth Edition(DSM IV).Exclusive criterion: parasuicider with idea or behavior of hurting oneself,but without intention to die.According to whether there was suicidal behavior or intention,116 patients were divided into 2 groups:attempted suicide group[n=52,35 males and 17 females,with an average age of (44± 13) years old] and no suicide group [n=64, 44 males and 20 females, with an average age of (48± 15) years old].Other 63 cases were taken as the normal controls[33 males and 30 females, with an average age of (55± 17) years old]. INTERVENTIONS:DNA of all the participants was extracted with the routine method of chloroform saturatedphenol leukocyte pheresis.The polymorphism of 5 HT2A receptor gene was analyzed with the restriction fragment length polymorphism. RESULTS:The G allele frequency of A 1438G polymorphism in the attempted suicide group(0.52) was significantly higher than that in the normal control group(0.39)(χ 2=3.91,P< 0.05).There were significant differences in the distribution of the 3 genotypes of AA, AG and GG between the attempted suicide group(0.23,0.50, 0.27) and normal control group(0.32, 0.59, 0.09)(χ 2=6.12,P< 0.05). CONCLUSION:Attempted suicide of psychiatric patient is associated with A 1438G polymorphism of 5 HT2A receptor gene.G allele may be a risk factor for the attempted suicide of psychiatric patient.

Translational medicine emerging as a new concept in medical field, concentrates on the integration of theory and practice , and the conjunction of basic medical science and clinical medicine. Barriers between the basic medicine and clinical medicine existing in medical education in China make students possess limited intelligence, narrow perspectives and low creativity etc., causing the medical teaching unable to conform to the demand of translational medicine. The article introduces the basic conception of translational medicine and the necessity of medical teaching reform, the meth-ods of promoting teaching reform by integrating the curriculum systems, bettering experimental teach-ing systems, strengthening humanities education, improving faculty construction and optimizing eval-uation system for students.

Objective To explore the dynamic change of serum leptin interleukin-6(IL-6),and interleukin-8(IL-8)during cerebral infarction,and to analyze the possible relationship between these factors and the severity of the disease.Method The levels of serum leptin,IL-6 and IL-8 were measured by radioimmunoassay in 60 patients with acute cerebral infarction,who were admitted to the hospital within 24 hours after stroke.Patients' neurologic impairment were evaluated by European Stroke Scale(ESS)at 1,3 and 7 days respectively after hospitalization.At the same time,another 30 normal adults were enrolled set as control group. Results The levels of serum Leptin,IL-8 in patients group after at 1 days,3 and 7 days were higher than those in the controls(P0.05).The ESS of these patients were 76.77?26.42,70.02?29.17,74.65?28.42 respectively,after 1,3 and 7 days.It shown that ESS simply correlated to the level of IL-8(P0.05).Statistic analysis was carried,but by using t test and linear correlation.(SPSS11.5 statistical package).Conclusion Leptin,IL-6 and IL-8 probably play a role in the inflammatory,reaction during acute cerebral infarction.The IL-8 may reflect the severity of the disease.

Serum glycosyl protein and glucose levels were concomitantly measured in 35 DACVD patients, 59 ACVD patients and in a control group of 65 healthy subjects. Our data revealed statistically significant differences in serum glucose between the ACVD group and control group (P0.05).Hyperglycosemia occurring in ACVD may be a response, or really means diabetes mellitus. Our data indicated that determination of serum glycosyl protein may be useful for the differential diagnosis of diabetic neuropathy from the reactive hyperglycosemia in ACVD.

Objective To observe the effect of electroacupuncture (EA) plus oxygenmedicine (OM) on the expression of Bcl-2 and Bax in the hippocampal CA 1 area in cerebral ischemia/reperfusion injury (CI/RI) rats. Methods Thirty SD rats were randomized into sham-operation,model,EA,OM,EA+OM groups (n=6/group). CI/RI model was established by using modified Pulsinelli 4 vessel occlusion and reperfusion. EA (100 Hz,3.5 mA) was applied to "Baihui" (GV 20) and "Zusanli" (ST 36) 30 min,once daily for 4 days. Rats of OM and EA+OM groups were put into a box filled with oxygen and atomized herbal medicines containing Bingpian (Borneolum),Shexiang (Moschus),Huangjing (Rhizoma Polygonati),Shouwu (Radix Polygoni Multiflori),etc. for 30 min,once daily for 4 days. Bcl-2 and Bax expression of the hippocampal CA 1 area was detected by immunohistochemistry. Results Compared with sham group,the numbers of Bcl-2 immunoreaction (IR) and Bax IR positive cells,and the immunoactivity of Bcl-2 IR and Bax IR positive products in the hippocampal CA 1 area were increased significantly in model group (P0.05). Conclusion EA and OM and EA+OM can effectively regulate the expression of Bcl-2 and Bax in the hippocampal CA 1 area in CI/RI rats,and the effects of EA+OM are significantly superior to those of simple EA and simple OM,which may contribute to their effect in improving cerebral ischemia.

BACKGROUND: Mild hypothermia (28-35 ℃) is becoming one of the promising methods in treating acute ischemic stroke. Hypothermia can effectively lessen brain edema, which is one of its neuroprotective roles.OBJECTIVE: To investigate the effect of mild hypothermia on brain water content and aquaporin-4 (AQP4) expression level following global cerebral ischemia-reperfusion injury in rats, so as to study the neuroprotective mechanisms of mild hypothermia.DESIGN: A randomized controlled trial.SETTING: Neurobiological Laboratory of Xuzhou Medical College.MATERIALS: 110 healthy male SD rats with body mass 250-300 g, provided by the Animal Center of Xuzhou Medical College, No. SYNK (Jiangsu) 2002-0079, were selected and randomly divided into 3 groups by SPSS 11.0software: ①sham-operated group (n=10);②normothermiagroup (n=50); ③mild hypothermia group (33 ℃, n=50). Normothermia group and mild hypothermia group were subdivided into five reperfusion subgroups for 6 hours, 1, 2, 3 and 7 days, respectively with 10 rats in each subgroup,in which 5 rats were used for measurement of brain water content, and others for HE staining and immunohistochemistry staining.METHODS: The models of global cerebral ischemia were established in the normothermia group and mild hypothermia group by four-vessel occlusion (4-VO) method with ischemia for 15 minutes as Pulsinelli described.The rats in the sham-operated group were only underwent the electrocauterization of bilateral vertebral arteries and the isolation of common carotid arteries except for occlusion of common carotid arteries. Twenty-four hours later, the rats were decapitated to take out the brains. The brains of rats in the normothermia group and mild hypothermia group were taken out to make sections for HE staining and immunohistochemistry staining, and the dynamic change of pathology of the brain tissue and AQP4 expression level were observed after reperfusion for 6 hours, 1, 2, 3 and 7 days, respectively. The brain wet-to-dry weight measurement was used to measure the brain water content of the rats at each time point of each group.MAIN OUTCOME MEASURES: ①The pathologic changes of brain tissues of rats in the normothermia group and mild hypothermia group. ②The brain water content and the AQP4 expression level of all rats at each time point.RESULTS: ①After 6 hours of reperfusion, brain edema appeared in the normothermia group including amplification of periyascular spaces and intercellular spaces, rarefaction of brain tissues, etc, which got worst after 2 days of reperfusion; the phenomenon of brain edema of the rats in the mild hypothermia group at each time point was relatively lighter than the normothermia group. ②Brain water content of the normothermia group and mild hypothermia group was increased after 6 hours of reperfusion and reached peak at the 2nd day; Although decreased at the 7th day, it was still higher than the sham-operated group. The brain water content of the mild hypothermia group at each time point was less than the normothermia group (values after 6 hour and 7 day, P ＜ 0.01, the rest groups P ＜ 0.05).③AQP4 expression level of the normothermia group and mild hypothermia group was increased after 6 hours of reperfusion and reached peak at the 2nd day. Although decreased at the 7th day, it was still higher than the sham-operated group.The AQP4 expression level of the mild hypothermia group at each time point was lower than the normothermia group (P ＜ 0.01).CONCLUSION:The change tendency of AQP4 level is parallel to that of brain water content after ischemia reperfusion, which indicates that the upregulation of AQP4 expression is one of molecular mechanisms for the for mation of ischemicbrain edema. Mild hypothermia can release ischemic brain edema by inhibiting AQP4 expression, which is one of its mechanisms.

Objective To investigate the location of receptor interacting protein 3( RIP3) in Necroptosis and its function in this signal passage, and explore the relationship between receptor interacting protein 1 ( RIP1 ) and RIP3 in nuclear translocation. Methods Primary cerebrocortical neurons were cultured for 12 days,then pre-treated with zVAD-fmk(20μ,mol/L) for half an hour to block apoptosis. ①Extracting nuclear and cytoplasmic protein after neurons were exposed to TNF for different time ,then protein levels of RIP3 were analyzed by western blot and immunofluorescence for qualitative observation;②In the following research,the neurons were treated with Nec-1 and shRlPl ,then the protein level of RIP1 and RIP3 with western blot were analyzed, cell viability were determined by measuring LDH levels. Results ①In signaling pathways of necroptosis, the protein level of RIP3 in cytoplasmic decreased gradually with prolonged TNF exposure, to the corresponding it rolled up in nucleus and a-chieved the peak in 12 hours of TNF treatment ( Cytoplasmic 0. 45 ± 0. 03 ,0. 41 ± 0. 02,0. 73 ± 0. 03 ,0. 90 ± 0.01,1.15 ±0.04,1.30 ±0.02,0.99 ±0.03,0.63 ±0. 03;Nucleus 0. 07 ±0.02,0. 26 ±0.02,0. 57 ±0. 02,0. 68 ± 0.02,0. 80 ± 0.01,0.92 ± 0.02,1.28 ± 0.03,0. 87 ± 0.02) (P < 0.01). ②Blocking the relationship between RIP1 and RIP3 with necrostatin-1 and shRIPl , nuclear translocation of RIP3 decreased and caused a great increase in cell viability( 1.00 ±0.05,0.39 ±0.03,0.50 ±0. 03) (P<0. 01). Conclusion RIP3 mainly locates in cy-tolymph of normal cells,it translocates into nucelus as necroptosis takes place. RIP1 function with RIP3 in nuclear translocation. Block nuclear translocation of RIP3 is a potential way to protect cells.

Objective To analyze the drug resistance genes and homology in nosocomial infection caused by carbapenem‐resistant Acinetobacter baumannii(CRAB) .Methods Forty strains of CRAB were collected and the minimal inhibitory concentration (MIC) was detected by the agar dilution method .The main carbapenemase genes were detected by PCR .The PCR products were performed the DNA sequencing .The enterobacterial repetitive intergenic consensus (ERIC)‐polymerase chain reaction(PCR) was performed to analyze the genotypes and homology of these strains .The plasmid conjugation experiment was carried out to investigate the trans‐mission of resistant genes .Results Forty strains of CRAB maintained the good sensitivity only to polymyxin B (100% ) and cef‐operazone/sulbactam(57 .5% ) .All of them had different degrees of resistance to other drugs .The OXA‐23 and OXA‐51genes were detected by PCR amplified in all of the CRAB strains .The homology analysis showed that the strains could be divided into 4 types according to 80% of cluster similarity coefficient ,the type A1 (19 strains) was the major epidemic strain ,moreover the various de‐partments had cross infection ;the plasmid conjugation experiment failed .Conclusion There is an endemic spread of CRAB in our hospital and its resistant mechanism is mainly related with OXA‐23 gene expression ;in the rational antibiotics use at the same time , the isolation and prevention measures should be done well for avoiding the hospital cross infection .