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 explore the preventive effects of simvastatin on ischemic-type biliary lesion (ITBL) after liver transplantation. Methods Totally 90 female SD rats,6 to 8 weeks old,weighing 200 to 250 g,were randomly divided into 3 groups,sham-operation group (n=10),control group (n=40,including 20 donors and 20 receipts),and experimental group (n=40,including 20 donors and 20 receipts). Rat liver transplantation was established by reconstructing hepatic artery. The rats of experimental group were fed with a normal chow containing 0.1% simvastatin for 1 week before operation and for 2 weeks after operation. The donor liver was stored in the University of Wisconsin solution at 4 ℃ for 12 h followed by transplantation. The animals were killed in 2 weeks after operation and the bile,bile duct and liver tissue were taken. High performance liquid chromatography (HPLC) was used to detect the concentrations of bile salts. Biochemistry,histopathology,immunohistochemistry and RT-PCR were employed to measure the level of phospholipids C,observe the pathological changes of liver,detect the protein and mRNA expressions of phospholipid transfer protein (Mdr2 P-glycoprotein). Results The biliary phospholipid composition of rats in the experimental group (3.874 8?1.414 8 mg/dl) was significantly increased,compared with the sham-operation group (1.982 1?0.413 5 mg/dl) and the control group (1.428 5?0.538 7 mg/dl) (P

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.

Objective To explore whether hypoxia-inducible factor-1α( HIF-1α )is involved in oxygenglucose deprivation (OGD)induced caspase-independent cell death in primary cortical neurons and whether their expression is infected by necrostatin-1 ( Nec-1 ).Methods ( 1 ) Primary cerebrocortical neurons were cultured for 14 days.Pretred z-VAD.Fmk (z-VAD)and Nec-1 with 0.1,1,5,10,25 and 50 μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours,then cell viability was determined by measure LDH level.(2)Pretred z-VAD before the neurons were exposed to OGD for 2 hours,then reoxygenated for 0,2,6,12,24and 48 hours.Then western blot analysis protein level of HIF-1 α ;rt-PCR check its RNA level.(3)Pretred z-VAD and Nec-1 with 25μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours.Then western blot analysis protein level of HIF-1 α; rt-PCR check its RNA level.Result ( 1 ) When cells were pretread Nec-1 with 5 μ mol/L(6.97 ± 0.06),the level of LDH was lower than cells untreated( 14.23 ± 0.08 ) (P＜ 0.05);At 25 μmol/L( 2.21 ± 0.05),the level of LDH was essentially the same as that of the control( 1.03 ±0.03 ) (P＞0.05).(2)The protein level of HIF-1 αwas different from normal (0.24 ±0.01 ) when exposed to OGD for 2 hours and reoxygenated for 2 hours (0.57 ± 0.09) and was highest after cells were exposed to OGD for 2 hours and reoxygenated for 12 hours(0.91 ± 0.08 ) (P＜ 0.05 ).The RNA level of HIF-1 α when cells were exposed to OGD was not deferent from normal (P ＞ 0.05 ).( 3 ) When cells were pretread with Nec-1 (0.32 ± 0.04 ),the protein level of HIF-1α were lower than untreated(0.83 ±0.03) (P＜0.05),but the RNA level of HIF-1α had no deference(P ＞ 0.05).Conclusion HIF-1α was involved in cell' s caspase-independent cell death;Nec-1 can protect neurons through inhibiting the expression of HIF-1α.

Objective To evaluate the efficacy of sequential psychotherapy combined with selective serotonin reuptake inhibitors(SSRI) s on refractory obsessive-compulsive disorder(OCD). Methods 50 patients with refractory OCD were given to sequential psycho therapy in the original basis of the SSRIs drugs treatment. The patients were given psychoanalysis,cognitive behavioral therapy and reality therapy for 8 weeks at the end of in the first stage, second stage and the third stage. The efficacy was assessed by the Yale-Brown Obsessive Compulsive Scale(Y-BOCS) before treatment,8 weeks, 16 weeks, and 24 weeks after treatment, respectively. Results The total score of the YBOCS before the treatment and at the 24th week after treatment was (21.10 ±4.69) and ( 10.18±6. 14). According to the score-reducing rate of the YBOCS , recovery rate was 34.0%, effective rate was 42.0%. Conclusion Sequential psychotherapy combined with SSRIs is an effective treatment for refractory OCD.

In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.
Cell Differentiation ; Cerebrospinal Fluid ; chemistry ; Culture Media ; chemistry ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; Neural Stem Cells ; cytology

Objective To explore the hearing screening, and the change and outcome of hearing impairment of high risk infants. Meth-ods From March, 2015 to March, 2016, 336 high risk infants were screened with otoacoustic emissions (OAE), auditory brainstem response (ABR) and brainstem auditory evoked-potential (BAEP) 0-1, 3, 6, 12 months after born, respectively. Results Among the 336 high risk in-fants, 29 failed the examinations within the 1st month, 37 cases failed in the 3rd month, 27 cases recovered in the 9th month, and 7 cases re-covered in the 12th month, 3 cases were finally diagnosed as deafness (0.89%). Conclusion OAE, ABR combining with BAEP examination may obtain comprehensive diagnosis of hearing impairment for high risk infants, continuous listening comprehension monitoring can effec-tively dynamically observe the hearing impairment, changes and outcome of high risk infants.

Objective To investigate the different expressions of pathological tissue and serum microRNAs (miRNAs)in Hirschsprung disease(HSCR). Methods Pathological colon tissues and serum samples were obtained from 52 confirmed HSCR cases respectively by surgery and pathology and from 52 matched controls,respectively. An initial screening of the tissues and serum microRNA expression were performed through TaqMan Low Density Array. The candidate tissue and serum miRNAs were validated by quantitative real - time - PCR in the 20 paired array samples and extra 32 paired samples after the integration of the screening result. The bioinformatical software online including miR-base,Target Scan,PicTar and MiRanda were used to predict the target mRNA of the consistent microRNAs in the tis-sues and the serum. Results Compared with the controls,47 microRNAs were differently expressed in HSCR tissues, including 17 up - regulated miRNAs and 30 down - regulated miRNAs;32 upregulated miRNAs were also detected to be differently expressed in the HSCR serum. Among these microRNAs,miR - 218 - 1 and miR - 885 - 5p were identi-fied to have a consistent significant different expression in both tissues and the serum,which were validated as high -expressed in microarray samples and expanded 32 paired samples(miR - 218 - 1:tissue array 0. 017 58 ± 0. 002 29 vs 0. 003 37 ± 0. 000 50,P ﹤ 0. 001;tissue expanded expression 0. 013 53 ± 0. 001 74 vs 0. 004 43 ± 0. 000 60,P ﹤0. 001. miR - 885 - 5p:tissue array 0. 000 30 ± 0. 000 11 vs 0. 000 04 ± 0. 0000 08,P = 0. 027 6;tissue expanded ex-pression 0. 004 59 ± 0. 000 16 vs 0. 000 04 ± 0. 000 01,P = 0. 014 5. miR - 218 - 1:serum array 0. 769 60 ± 0. 285 50 vs 0. 045 14 ± 0. 015 07,P = 0. 015 5;serum expanded expression 1. 151 00 ± 0. 430 00 vs 0. 023 07 ± 0. 003 81,P =0. 008 7. miR -885 -5p:serum array 1. 595 00 ±0. 441 70 vs 0. 169 40 ±0. 034 46,P =0. 001 2;serum expanded expres-sion 1. 689 00 ±0. 453 00 vs 0. 146 10 ± 0. 031 24,P = 0. 001 2). Specifically,the target genes of these 2 microRNAs were RET,PLAG1 and NeuroD1,which had been reported to be directly related to HSCR. Conclusions Significantly dif-ferential expressed miRNAs exist in the pathological tissue and the serum of HSCR. MiR - 218 - 1 and miR - 885 - 5p, which showing consistent differential expression,may be involved in the pathogenesis of HSCR.

Objective To investigate MRI appearances of aquaporin(AQP) and its effect in different brain regions of patients with Parkinson's disease(PD).Methods A prospective study was carried out in 33 PD patients(PD group) and 23 gender-and age-matched healthy controls (control group).Clinical data of PD patients were collected.The aquaporin imaging of diffusion-weighted magnetic resonance imaging (MRDWI) with multiple b-values in different brain regions were performed to detect the apparent diffusion coefficient(AQP-ADC) values of aquaporin.The PD patients were assessed and graded by modified Hoehn-Yahr grading,then the AQP-ADC values of control group,mild PD group,moderate and severe PD group were analyzed using one-way analysis of variance.The correlation analysis was carried out to detect the relationship between AQP-ADC values in different brain regions and Hoehn-Yahr grading of PD patients.Results Compared with control group,mild PD group had significantly higher AQP-ADC values in red nucleus(RN) and globus pallidus(GP) ((0.24±0.04) vs (0.21±0.04),(0.21±0.04) vs (0.16±0.04);both P＜0.05);while the AQP-ADC values in RN and GP of moderate and severe PD group were significantly lower than that of mild PD group((0.21±0.02) vs (0.24±0.04),(0.18±0.03) vs (0.21±0.04);both P＜0.05);but there was no significant difference between moderate and severe PD group and control group(P＞0.05);and there was also no significant difference in substantianigra (SN),putamen (Pu) and thalamus (THA) among control group,mild PD group and moderate and severe PD group(P＞0.05).The correlation analysis showed that there were negative correlations between the AQP-ADC values in RN and GP and Hoehn-Yahr grading(r=-0.479 and-0.395,P＜ 0.05),while there was no correlation in SN,Pu and THA (P＞ 0.05).Conclusion The AQPADC values are increased in RN and GP of mild PD patients,and decreased in moderate and severe PD patients,while there is no significant change in SN,Pu and THA of the two groups,suggesting that the expression of AQP in different brain regions may be related to the severity and pathological stage of PD.

Objective:To identify the epitope of mAb15A11 which is specific against RA associated autoantigen cartilage oligomeric matrix protein ( COMP ).Methods: A filamentous phage library displaying random linear dodecapeptides was used to mapping the epitope of mAb15A11.After three rounds of screenings,40 phage clones were selected at random and sequenced.The specificity of phages was confirmed by enzyme immunoassays.Homology search by ClustalW2 and structure analysis by PyMol to identified the epitope amino acid sequence.Western blot analysis of COMP and ELISA analysis of COMP-derived peptides were used to confirm epitope′s characterization.Results: After repeated screenings using bio-panning method, 2 clones were identified, which interacted specifically with mAb 15A11.Homology search did not find succession consensus sequence within COMP molecular,which indicated that the epitope was not linear.PyMol Structure analysis identified the rationality of conformational epitope.Western blot analysis and ELISA of EDTA-treated COMP further prove an conformational structure of the epitope recognized by mAb 15A11.ELISA analysis of COMP-derived peptides demonstrated both disulfide bonds between 229 C-243 C and 237 C-253 C and every epitope amino acid of 232 G,238 H,240 H,241 A,244 V,247 R and 251 R were essential to the binding of mAb 15A11 with COMP.Conclusion: In this study, the potential B cell antigentic epitopes of mAb 15A11 was identified by phage display library.The epitope amino acids sequence and char-acterization were also recognized.It may have important theoretical value for the study of reaction mechanism of COMP antibody and antigen and may also show application significance in the detection of rheumatoid arthritis.