Objective:To investigate the effective mechanism for transfection of rat brain derived neurotrophic factor(BDNF) gene by recombinant adeno-associated virus in the cultured hippocampal neurons.Methods:rAAV-BDNF was injected into the serum free-induced neurons.DAPI,PI and Actin stain was performed to measure the apoptosis cells and morphogenesis of the cells.Results:After the gene transfer,the survival rate of the serum free-induced neurons was increased by 65%.Conclusion:The recombinant adeno-associated virus vector can protect the serum free-induced neurons from Apoptosis.

Objective To construct eukaryotic expressing vector of the mouse NMNAT1 (nicotinamide mononucleotide adenylyl-transferase) gene and examine its ability to express the NMNAT1 gene in Hela cells.Methods The full-length NMNAT1 cDNA sequence was amplified by PCR and cloned into the plasmid of T-vector and then to pcDNA3.1 construct.The recombinant plasmid pcDNA3.1-NMNAT1 was identified by DNA sequencing and then transfected with Lipofectamine2000 into Hela cells.The expression of NMNAT1 was detected by real time quantitative PCR (qPCR) and Western blot after 48 h transfection.Results The recombinant eukaryotic vector carrying NMNAT1 gene was constructed successfully in a match with database and this vector could up-regulate the expression of the NMNAT1 gene both in mRNA and protein levels in Hela cells.Conclusions The eukaryotic vector carrying NMNAT1 gene (pcDNA3.1-NMNAT1) enhances the expression of NMNAT1 gene.

Objective To discuss the clinical significance of serum pyridinoline cross-linked carboxy-terminal telopeptide of typeⅠcollagen in patients with rheumatoid arthritis (RA). Methods Serum ⅠCTP was measured by ELISA and RF, CRP by immunonephelometry assay in 55 patients with active and 25 with stable RA as well as 52 age and sex-matched normal controls. Results SerumⅠCTP was positively correlated with RF and CRP (rs=0.407, 0.534, P0.05). The sensitivity, specificity and accuracy of serumⅠCTP test were 67.3%, 84.6%, 75.7% in serum of the patients with active RA respectively. The mean serumⅠCTP in normal controls was (3.5?2.0) ?g/L. The serumⅠCTP was more sensitive than CRP in the patients with active RA. Conclusion SerumⅠCTP in patients of RA is a reliable assistant marker for early diagnosis, monitoring of the course and the effectiveness of treatment.

Objective:To investigate the influencing factors for synchronous colorectal liver metastasis (synCRLM).Methods:The retrospective case-control study was conducted. The clinicopathological data of 3 172 patients with primary colorectal cancer (CRC) who were admitted to the Affiliated Hospital of Qingdao University from January 2010 to January 2016 were collected. There were 1 946 males and 1 226 females, aged (63±12)years, with a range from 21 to 97 years. Observation indicators: (1) general data analysis; (2) clinicopathological data analysis; (3) analysis of influencing factors for synCRLM. Measurement data with normal distribution were represented as Mean±SD. Count data were represented as absolute numbers. The influencing factors for synCRLM were analyzed after excluding missing data of tumor differentiation degree, tumor diameter, pathological T stage and N stage. Univariate analysis was conducted by chi-square test or Logistic regression model. Multivariate analysis was conducted by Logistic regression model. Results:(1) General data analysis: among the 3 172 patients, cases with age ≤29 years, from 30 to 39 years, from 40 to 49 years, from 50 to 59 years, from 60 to 69 years, from 70 to 79 years, and ≥80 years were 15, 82, 342, 774, 965, 759 and 235, respectively. There were 2 972 patients in Qingdao, 172 cases in Yantai and 28 cases in Weihai. Of the 2 972 patients in Qingdao, there were 422 cases in Shinan District, 658 cases in Shibei District, 457 cases in Huangdao District, 144 cases in Laoshan District, 188 cases in Licang District, 205 cases in Chengyang District, 252 cases in Jimo District, 221 cases in Jiaozhou City, 255 cases in Pingdu City, 170 cases in Laixi City. (2) Clinico-pathological data analysis: among the 3 172 patients, there were 1 639 cases of colon cancer including 972 cases with left colon cancer and 667 cases with right colon cancer, 1 533 cases of rectal cancer. There were 2 981 cases of adenocarcinoma, 165 cases of mucinous adenocarcinoma, 10 cases of signet ring cell carcinoma and 16 cases of other types including carcinoid tumor, squamous carcinoma, tubular adenocarcinoma, etc.There were 162 cases with highly differentiated adenocarcinoma, 5 cases with highly-moderately differentiated adenocarcinoma, 2 338 cases with moderately differentiated adenocarcinoma, 80 cases with moderately-poorly differentiated adeno-carcinoma, 396 cases with poorly differentiated adenocarcinoma and 191 cases missing tumor differentiation data. There were 708 cases with tumor diameter <4 cm, 1 957 cases with tumor diameter ≥4 cm and 507 cases missing tumor diameter data. There were 486 cases in T1 or T2 stage of pathological T stage, 2 169 cases in T3 or T4 stage of pathological T stage and 517 cases missing tumor pathological T staging data. There were 1 563 cases in N0 stage of pathological N staging, 1 062 cases in N1 or N2 stage of pathological N staging and 547 cases missing tumor pathological N staging data. There were 2 895 cases without synCRLM and 277 cases with synCRLM. There were 2 799 cases without diabetes and 373 cases with diabetes. There were 2 931 cases without fatty liver and 241 cases with fatty liver. There were 2 989 cases negative for hepatitis B surface antigen (HBsAg) and 183 cases positive for HBsAg. (3) Analysis of influencing factors for synCRLM. Results of univariate analysis showed that gender, tumor location, tumor differentiation degree, tumor diameter, pathological T stage, fatty liver, HBsAg were related factors for synCRLM in primary colorectal cancer ( χ2=7.400, 7.577, 7.111, 4.513, 12.125, 5.686, 5.919, P<0.05), and neutrophils counts, lymphocytes counts, platelet counts, alanine aminotransferase (ALT), aspartate aminotrans-ferase (AST), total bilirubin, γ-glutamyltransferase (GGT), triacylglycerol (TG), total cholesterol (TC), carcinoembryonic antigen (CEA), and CA19-9 were related factors for synCRLM in primary colorectal cancer ( odds ratio=1.101, 0.807, 1.002, 1.017, 1.023, 1.027, 1.012, 0.686, 1.169, 1.007, 1.004, 95% confidence interval as 1.048-1.156, 0.678-0.960, 1.001-1.004, 1.011-1.024, 1.016-1.031, 1.011-1.044, 1.009-1.015, 0.541-0.869, 1.047-1.306, 1.006-1.008, 1.003-1.004, P<0.05). Results of multivariate analysis showed that cases as male, case with positive HBsAg, AST, GGT, TC, CEA and CA19-9 were independent risk factors for synCRLM in primary colorectal cancer ( odds ratio=1.503, 2.492, 1.018, 1.007, 1.301, 1.005, 1.003, 95% confidence interval as 1.038-2.178, 1.443-4.304, 1.003-1.034, 1.003-1.011, 1.112-1.522, 1.003-1.006, 1.002-1.003, P<0.05), and lymphocytes, ALT and TG were independent protective factors for synCRLM in primary colorectal cancer ( odds ratio=0.777, 0.983, 0.602, 95% confidence interval as 0.608-0.993, 0.966-0.999, 0.421-0.862, P<0.05). Conclusion:Cases as male, case with posotive HBsAg, AST, GGT, TC, CEA and CA19-9 are independent risk factors for synCRLM in primary colorectal cancer, while lymphocytes, ALT and TG are independent protective factors for synCRLM in primary colorectal cancer.

Objective To compare the efficacy of amisulpride and other SGAs in treating the negative symptoms of Schizophrenia.Methods The randomized controlled trials (RCTs) about Schizophrenia treated with amisulpride and other SGAs from Jan 1995 to Mar 2013 were searched in The Pubmed,EMbase,Cochrane Library,WanFang Data,CNKI and VIP.Two reviewers independently screened the literatures,extracted the data,and evaluated the methodological quality.Than meta-analyses were conducted by using RevMan 5.1 and Stata 12.0 software.Results The totall3 RCTs were included.Among the 1814 patients involved.The results of meta-analyses showed that the score of PANSS-N was no significant differences between two groups (MD =-0.33,95％ CI:(-0.87,0.21),Z =1.20,P =0.23) ; and the score of SANS was no significant differences between two groups (MD =-0.21,95％ CI:(-1.51,1.50),Z =0.31,P =0.76).The side effects were more in other SGAs group than those in amisulpride group.Conclusion Amisulpride is as effective as other SGAs for the treatment of schizophrenia with predominantly negative symptoms,and it has more advantage than other SGAs in safety.

Objective To study the imaging features of sacroiliac joints(SIJ) in patients who were misdiagnosed as spondyloarthritis(SpA).Methods A total of 34 patients with chief complaint of back pain and misdiagnosed as SpA from January 2007 to April 2013 in Department of Rheumatology Chinese PLA General Hospital were enrolled.The imaging,clinical manifestations,laboratory examinations data were analyzed.Result The main reason for misdiagnosis as SpA was because of sacroiliitis presenting on imaging.The final diagnoses included 24 patients as SIJ infection,4 patients as neoplastic diseases,2 patients as metabolic bone diseases,2 patients as sacroiliac joint degeneration,1 patient as gout of sacroiliac joint,1 patient as diffuse idiopathic bone hypertrophy.For patients with infection,there were 10 patients receiving X-ray and 22 patients receiving CT of SIJ.However,5 and 7 patients had negative results respectively.These patients with infection had abnormalities in MRI including all with bone marrow edema,21 patients with erosion of bone and joint,22 patients with muscle involved.As to the patients with malignancies,SIJ CT scan appeared normal.Bone marrow edema and erosion in MRI were found in all neoplasm patients expect one as ependymoma.Adjacent muscles were involved in the patient with Ewing's sarcoma.Either X-ray or CT in other patients demonstrated obvious abnormalities,but only mild erosion of bone was found in MRI.Conclusion Bone marrow edema of SIJ in MRI represented not only in patients with SpA.Rheumatologists should analyze the clinical manifestations and laboratory examinations comprehensively in order to avoid the misdiagnoses.

Objective To compare concurrent chemoradiotherapy and sequential therapy effect on serum MMP-2 and TGF-β1 in local advanced non-small cell lung cancer (NSCLC).Methods From 2010 January to 2012 December,64 Ⅱ B and Ⅲ B stage patients with pathologically confirmed NSCLC were randomly divided into concurrent chemoradiotherapy group (group A) and sequential therapy group (group B).Each group had 32 patients.Group A was treated with three-dimensional conformal radiotherapy and concurrent chemotherapy with TC or EP.Group B received TC or EP regimen chemotherapy after three-dimensional conformal radiotherapy.Serum MMP-2 and TGF-β1 on those patients from preradiotherapy,radiotherapy in one month to post-treatment were measured by enzyme-linked immunoabsorbent assay.The dynamic changes of MMP-2 and TGF-β1 were compared.Results The remission rates in groups A and B were 90.6 ％ and 68.8 ％,the effective rate of treatment in group A was better than that of group B (x2 =4.730 0,P =0.029 6).The long-term effect analyzed with Kaplan-Meier method,the median time to tumor progression (TTP) were 9.1 months and 8.2 months,there was no statistically significant difference (P =0.100 3).The overall survival rates between two groups after the Log-rank test had significant difference (P =0.048),the median survival time (MST) were 17.8 and 15.9 months,1 year OS rates were 65.05 ％ and 60.24 ％,2 years OS rates were 49.45 ％ and 43.07 ％.The MMP-2 level of A group and B were (276.5±98.2) μg/ml and (263.9±103.5) μg/ml,there was no significant difference (t =0.499 6,P =0.619 1) before radiotherapy,they were (242.1±53.2) μg/ml and (298.7±68.4) μg/ml after radiotherapy,there was significant difference (t =3.694 9,P =0.005) and after treatment were (60.5 ±24.4) μg/ml and (75.2±30.7) μg/ml,there was significant difference (t =2.120 5,P =0.038 0).The TGF-β1 level of A group and B were (1 624.3±454.2) ng/ml and (1 564.9±517.8) ng/ml,there was no significant difference (t =0.208 6,P =0.835 4) before radiotherapy,they were (1 383.5±469.3) ng/ml and (1 785.3±412.6) mg/ml after radiotherapy,there was significant difference (t =3.637 3,P =0.006 0) and after treatment were (610.5±215.4) ng/ml and (750.3±263.7) ng/ml,there was significant difference (t =2.322 6,P =0.023 5).Conclusions Concurrent chemoradiotherapy could effectively antagonize radiation-induced MMP-2 and TGF-β1 expression increased in locally advanced NSCLC.This study suggests that the concurrent chemoradiotherapy can inhibit abilities of tumor invasion and metastasis through decreasing the MMP-2 and TGF-β1 levels.

Objective To summarize the progress on the injury mechanism of vascular endothelial cells in atherosclerosis.Methods The latest progress was reviewed in recent literatures.Results All kinds of etiological factors have activated NF kappa B and cytokines in the development of atherosclerosis, which lead to expression of cell adhesive molecules and adhesion of monocytes to vascular endothelial cells.A variety of inflammatory mediums are released, which can directly damage endothelial cells.Besides, the inflammatory mediums make monocytes and neutrophils attach to endothelial cells by immune mechanisms, which injure the endothelial cells more severely. Meanwhile the damaged membrance structure leads to the production of AECA which activates the complementary system. Then the vascular endothelial cell injury is aggravated and the development of atherosclerosis accelerated. Conclusion It is very important to recognize the injury mechanism of vascular endothelial cells in the development of atherosclerosis for prevention and treatment of atherosclerosis.

Objective To investigate the expression of WNK1 and WNK4 genes in the mouse kidney. Methods The probes corresponding to the genes were amplified by RT PCR from mouse kidney, and then Northern blotting was performed on mouse multiple tissues membrane. The fragments were cloned into pGEM T vector,the correct plasmids were linearized and transcripted into RNA probes in vitro. In situ hybridization was performed on the mouse kidney sections. Results WNK1 gene was widely expressed in mouse tissues. There was a 9 5 kb strong signal in kidney. WNK4 gene was mainly expressed in kidney and had a 4 4 kb signal. In situ hybridization showed WNK1 gene was mostly expressed in the distal convoluted tubule whereas WNK4 gene was expressed in the medullary collecting ducts besides the distal convoluted tubules. Conclusion Both WNK1 and WNK4 genes were expressed in kidney. WNK4 was expressed more widely than WNK1 in mouse kidney.

Objective:To investigate the characteristics of perceived stress and coping style and their relationship withsymptoms in college students with psychosis-risk syndrome (PRS).Methods:A two-stage screening process was conducted in 3160 college students.Participants were screened with 16-Item Version of the Prodromal Questionnaire (PQ-16).The students with scores of PQ-16 ≥9 were further interviewed with Structured Interview for Prodromal Syndromes (SIPS).Twenty-two individuals who met the criteria of PRS on SIPS were enrolled as the PRS group.The control group (n =22) were randomly selected from the participants with scores of PQ-16 ＜9 who were matched with age,gender and education level.Two groups were assessed with Chinese Perceived Stress Scale (CPSS),Trait Coping Style Questionnaire (TCSQ) and Self-rating depression scale (SDS) to measure subjective stress,coping style and depressive symptoms.Results:PRS group reported higher CPSS total scores [(34 ± 8)vs.(22 ± 8)] and negative coping styles scores [(35 ± 5)vs.(28 ± 7)],and lowerTCSQ positive coping styles scores [(31 ±6)vs.(37 ±4)] than the controls (Ps ＜0.05).The CPSS scores in PRS group were positively correlated with the scores of negative coping style (r =0.57,P ＜0.01),and the SDS scores were positively correlated with CPSS total scores and scores of negative coping style (r--0.86,0.66,Ps ＜ 0.01).There was no significant relation of psychosis-risk symptoms to scores of CPSS and TCSQ (P ＞0.05).Conclusion:It suggests that college students with PRS have a higher perceived stress and tend to use more negative coping style,which may be positively correlated with depression.