Objective:To identify the differentially expressed long-chain non-coding RNA(lncRNA) and mRNA using ribonucleic acid sequencing(RNA-seq), and construct a competing endogenous RNA(ceRNA) regulatory network in mice with sepsis-associated encephalopathy.Methods:Ten clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups( n=5 each) using a random number table method: sham operation group(group Sham) and sepsis group(group Sepsis). Sepsis was induced by cecal ligation and puncture(CLP) in group Sepsis, while group Sham only underwent laparotomy without CLP. Morris water maze test and contextual fear conditioning test were performed to detect the cognitive function on 1 day before CLP and 3 days after CLP. Three mice were randomly sacrificed in group Sham, and 3 mice with the worst results in the cognitive function test were sacrificed in group Sepsis. The hippocampal tissues were obtained for RNA-seq via the BGISEQ-500 platform, and the differentially expressed mRNA and lncRNA were identified. The differentially expressed mRNAs and lncRNAs were visualized and analyzed by Dr. Tom platform provided by Shenzhen BGI Technology Service Co., Ltd., and the ceRNA regulatory network was constructed using the online visualization tool Cytoscape software. Results:Compared with group Sham, the escape latency was significantly prolonged, and the percentage of time of staying at the target quadrants and percentage of time spent freezing were decreased in group Sepsis( P<0.05). A total of 62 differentially expressed lncRNAs were obtained from RNA-seq, of which the expression of 45 lncRNAs was up-regulated and the expression of 17 lncRNAs was down-regulated.There were 282 differentially expressed mRNAs identified from RNA-seq, of which the expression of 173 mRNAs was up-regulated, and the expression of 109 mRNAs was down-regulated.Gene Ontology enrichment analysis revealed that the differentially expressed mRNAs were involved in biological processes such as memory, learning or memory, inflammatory responses, regulation of aging-related behavioral decline, and regulation of synaptic plasticity. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that differentially expressed mRNAs were enriched in IL-17 signaling pathway, TNF signaling pathway, NF-κB signaling pathway and etc. KDA analysis was performed on the differentially expressed mRNAs to identify the key driver genes, and the results showed that Ch25h, Il6ra, Lcn2, Sgk1, Nr4a3, Osm, Saa3, Ccl7, Sqle, Dhcr24 were the key SAE genes.A competing endogenous RNA regulatory network was successfully constructed based on 9 lncRNAs, 28 mRNAs and 134 miRNAs in the hippocampus of mice with SAE. Conclusions:The results of RNA-seq find that 10 mRNAs including Ch25h, Il6ra, Lcn2, Sgk1, Nr4a3, Osm, Saa3, Ccl7, Sqle, Dhcr24 and lncRNAs such as Rian, Gm35874 and Gm34347 are key genes regulating SAE in mice. Meanwhile, a ceRNA regulatory network based on lncRNA-miRNA-mRNA is successfully constructed in the hippocampus of mice with SAE.

Peripheral nerve injury (PNI) is an important clinical complication, which brings long-term physical and psychological pain and economic burden to patients. There is no satisfactory treatment plan for PNI. Although microsurgery technology has been greatly developed, some peripheral nerve defects or ruptures caused by external forces can be repaired by surgery or nerve transplantation. However, due to the weak ability of nerve cell regeneration and surgical operations may cause damage to the injured nerves, the patient's functional recovery may not be able to achieve the desired effect. Therefore, it is urgent to find a safe and effective method to treat PNI. Mesenchymal stem cells have special differentiation potential and can differentiate into a variety of cell types in vitro and in vivo, and have received widespread attention from researchers. In this paper, the research progress of mesenchymal stem cells in nerve injury repair was summarized, and the characteristics, functions of mesenchymal stem cells and the mechanism of action in peripheral nerve injury repair were reviewed.

Objective: To evaluate the role of DNA methylation in sepsis-associated encephalopathy in mice. Methods: A total of 144 clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups (n=36 each) using a random number table method: sham operation group (group Sham), sepsis group (group Sepsis), sham operation plus S-adenosyl methionine (SAM) group (group Sham+ SAM) and sepsis plus SAM group (group Sepsis+ SAM). Sepsis was produced by cecum ligation and puncture (CLP). In Sham+ SAM and Sepsis+ SAM groups, DNA methylated methyl donor SAM 100 mg/kg was intraperitoneally injected at 1 h before operation and 12 h after operation, while the equal volume of normal saline was given instead in Sham and Sepsis groups.The cognitive function was assessed using Y-maze and contextual fear conditioning test at 1, 3 and 7 days after CLP.Mice were sacrificed at 1, 3 and 7 days after CLP, and the hippocampal tissues were taken for determination of genome-wide DNA methylation (by colorimetric assay) and expression of DNA methyltransferase enzymes (DNMT1, DNMT3a, and DNMT3b), ten-eleven translocation (TET) enzymes (TET1, TET2 and TET3) and thymine-DNA glycosylase (TDG) mRNA (by fluorescent quantitative real-time). Results: Compared with group Sham, the time of staying at the novel arm was significantly shortened, the percentage of time spent freezing and the total number of entries into each arm were reduced, genome-wide DNA methylation in hippocampal tissues was decreased at 1, 3 and 7 days after CLP, the expression of DNMT1, DNMT3a, TET1, TET2, TET3 and TDG was up-regulated, and the expression of DNMT3b was down-regulated in group Sepsis (P<0.05). Compared with group Sepsis, the time of staying at the novel arm was significantly prolonged, the percentage of time spent freezing and the total number of entries into each arm were increased, the expression of DNMT1, DNMT3a, TET1, TET2, TET3 and TDG mRNA was down-regulated, and the expression of DNMT3b was up-regulated in group Sepsis+ SAM(P<0.05). Conclusion DNA methylation is involved in the development of sepsis-associated encephalopathy in mice.

Objective To investigate the prognostic value of preoperative neutrophil lymphocyte ratio (NLR) in patients with gastrointestinal stromal tumors (GIST).Methods The clinical-pathological data from 85 GIST cases were collected from the Second Affiliated Hospital of Wenzhou Medical University and retrospectively analyzed from June 2009 to December 2013.According to the exclusion criteria,79 cases were enrolled.Preoperative neutrophil and lymphocyte count was collected and NLR was calculated.According to the receiver operating characteristic (ROC) curve of NLR,GIST patients were divided into low NLR group (NLR ＜ 2.30) and high NLR group (NLR ≥ 2.30).Clinic-pathological features and five year disease free survival (DFS) were compared between the two groups.Results There was a statistical significant difference in tumor size and tumor risk between high NLR group and low NLR group (respectively,x2 =9.517,12.411,all P ＜ 0.05).Univariate analysis showed that the five year disease free survival rate of low NLR and high NLR group were 78％ and 32％ (x2 =18.749,P =0.000).By multivariate analysis a high NLR was identified as an independent risk factor of poorer prognosis for patients with GISTs (RR:3.516,95％CI:1.453-8.506,P=0.005).Conclusion A high preoperative NLR is an independent risk factor for the prognosis of GISTs.

Objective To investigate the clinical value of serum lipoprotein ( a ) concentration in evaluation of plaques characteristics for patients with coronary atherosclerotic heart diseases ( CAHD ) . Methods Using case-control method, Patients with suspicious CAHD, received coronary computed tomography angiography in the Shanghai East Hospital during October 2013 to June 2015 were enrolled.According to the results of coronary artery CTA, the patients were divided into two groups : the CAHD group (352 cases) and control group(438 cases) , the particle concentrations and mass concentration of lipoprotein(a), triglyceride, total cholesterol, HDL-C, LDL-C, glucose, HBA1c and hs-CRP and other tests were measured, the patients of CAHD group were divided into three subgroups by characteristics of coronary artery plaques including soft plaque (176 cases), calcified plaque (90 cases) and mixed plaque (86 cases), analysis were made with all these data.Using T test or variance analysis to compare the means between or among groups, the risk for CAHD was analyzed by logistic regression, the relationship between LP (a) -P and LP( a) -M were explored by linearly egression analysis, Conformance test were analyzed using kappa test.Results Compared with control group, the mean results of the CAHD group are significantly higher than that of control group, including LP (a) -P 18.5(8.3 -43.0))nmol/L vs.13.6 (7.6-32.4)nmol/L( t =-2.110), LP(a)-M 183(71 -361)mg/L vs.126(67 -293)mg/L(t =-2.063), age (62 ±9)years vs.(52 ±9)years(t=-7.691), hs-CRP 0.86(0.44-1.97) )mg/L vs.0.70(0.38-1.64)mg/L(t=-2.236), glucose (6.1 ±2.29 )mmol/L vs.(5.36 ±1.32 )mmol/L(t=-4.914), BA1c (6.13 ±0.98) % vs.(5.81 ±0.58) %(t=-4.842), APO(B) (1.09 ±0.33) g/L vs.(1.03 ±0.29) g/L( t=-2.407), all of the P values <0.05;The relative risk(RR)of age, glucose, LP( a)-P and LP ( a)-M are 1.067, 2.377, 1.384 and 1.342 respectively; Among the three types of plaques groups,the mean differences of age, TC, HDL-C, LDL-C and LP ( a)-P are statistically significant ( F=6.276,3.060,3.127,4.723,2.878;all of the P<0.05);The median of LP ( a)-P in the soft plaque group 20.3(8.3-48.2)nmol/L is higher than that of the mixed plaque group 15.7(7.3-26.0)nmol/L(P<0.05 ) and calcified plaque group 15.6 ( 8.1 -23.1 ) nmol/L ( P <0.05 ).The linearly regression equation of LP ( a) -M and LP( a)-P is Y=6.646X, r=0.939; Consistency test indicate the two methods are not consistent when used for grouping ( Kappa value is 0.557 ).Conclusions Serum concentration of lipoprotein(a) is an independent risk factor of CAHD, and the particle concentration of LP(a) is closely related to the characteristics of the plaques, especially to the soft plaque.

Objective To explore the serum level of Glucagon like peptide-1 in late-onset Alzheimer′s patients and its clinical significance.Methods Case control study.Collecting cerebral vascular disease fifty-five cases, diagnosed with late-onset Alzheimer′s disease sixty-one cases, type 2 diabetes mellitus fifty-one cases , type 2 diabetic patients combined with late-onset Alzheimer′s disease thirty-seven patients from the Shanghai East Hospital and partly Pudong area elderdly hospital during October 2013 to March 2014, and forty healthy persons as normal control from physical examination center of Shanghai East Hospital during September 2013 to February 2014.Measuring the concentrations of GLP-1,β-amyloid, Tau protein and other routinely used clinical tests in the serum of patients from the normal controls , cerebrovascular disease , late-onset Alzheimer′s disease, type 2 diabetes and type 2 diabetes mellitus combined with late-onset Alzheimer′s disease by ELISA method developed in our laboratory.The blood samples were also collected at three fixed time including fasting time ,1 hour after taking glucose , 2 hour after taking glucose, the concentrations of GLP-1 were determined in the LOAD group , T2DM group and the T2DM combined with LOAD group and normal control group.The concentrations of serum GLP-1 among groups were compared with single factor analysis of variance , and the concentrations of serum GLP-1 between the two groups were compared using LSD-t test.Analysing the correlation between GLP-1 and other indicators with Pearson analysis.Results The fasting GLP-1 levels of LOAD group were ( 123.4 ±20.8 ) nmol/L, and they were highest between the normal control group (78.6 ±6.0) nmol/L and the cerebral blood vessel disease group(89.0 ±8.7)nmol/L (F values were 3.46 and 1.98, P<0.05).The fasting GLP-1 levels of T2DM combined with LOAD group (157.9 ±28.6) nmol/L were higher than the LOAD group (123.4 ± 20.8) nmol/L (t =1.63,P <0.05), but there were no difference of the fasting GLP-1 levels between T2DM combined with LOAD group (157.9 ±28.6) nmol/L and T2DM group(153.8 ±18.0)nmol/L(t=0.96,P>0.05).Deficient secretion of GLP-1 after taking glucose 1 hour in most of the patients of T2DM combined with LOAD group (99.1 ±14.2) nmol/L, LOAD group(73.9 ±6.6 ) nmol/L and T2DM group (96.3 ±7.0 ) nmol/L could be concluded .The GLP-1 levels of T2DM combined with LOAD group after taking sugar 2 hour were (115.4 ±18.6)nmol/L ,and were higher than that of normal levels (63.3 ±6.2) nmol/L after taking sugar 2 hour(t=4.49,P<0.05).There were no difference between the GLP-1 levels of the LOAD group (73.6 ±5.8 )nmol/L and the GLP-1 levels of the normal group(63.3 ±6.2)nmol/L after taking sugar 2 hour (t=0.94,P>0.05).Pearson correlation analysis showed that the relationship of the levels of GLP-1 with Aβ( 1-42 ) and the levels of GLP-1 after taking glucose 1 h and 2 h were positively relative, and its coefficients of correlation were 0.401,0.436,0.722.Conclusions LOAD and T2MD are similar, and they have GLP-1 secretion shortage phenomenon after taking glucose , so monitoring dynamic change of GLP-1 after taking glucose may contribute to the auxiliary diagnosis of LOAD.

Objective To compare the difference of kidney function evaluated by using 3 different estimated glomerular filtration rate(GFR) equations in populations .Methods Retrospectively analyzed 65 856 patients who measured serum creatinine and Cysta‐tin C at the same time ,and come from the outpatients or inpatients of the hospital .The estimated GFR (eGFR) were calculated through 3 equations ,then compared the eGFR in the population and among different groups according to different kidney functions , and then grouped the people enrolled in the study again according to the eGFR calculated by using the 3 different equations and compared the differences among groups .Results Compared with the eGFR calculated by using Creatinine equation ,the correlation coefficients of the eGFRs calculated by using the other two equations were 0 .81 and 0 .90 ,respectively ,both P<0 .05 ;The differ‐ence between the means of eGFR were 6 .19 and 1 .79 mL/(min × 1 .73 m2 ) respectively with obvious significance (P<0 .01) ,in consistency analysis .There were obvious overestimation of kidney function when using Creatinine equation to calculate eGFR .Con‐clusion There is consistence and obvious difference by using the 3 CKD‐EPI′s eGFR equations .Physicians should choose suitable equations to evaluate kidney function in different populations .

Objective To evaluate the effect of neoadjuvant chemotherapy on lymph node micrometastasis and DC cell number in stage Ⅲ gastric cancer patients.Methods Lymph node micrometastases in pathologically negative lymph nodes from 76 stage Ⅲ gastric cancer patients [56 cases with neoadjavant chemotherapy (NCT) and 20 cases without],were assessed by cytokeratin-20 antibody.Dendritic cells (DC) in HE positive lymph nodes,HE negatives near tumor foci and HE negatives far foci were assessed by CD83 antibody respectively.Patients were divided into four groups:group 0 (20 cases without NCT),group 2 (two courses of NCT in 29 patients),group 3 (three courses of NCT in 16 patients),group 4 (four courses of NCT in 11 patients).Results In NCT group there were two patients with micrometastases (2/56).All the patients were with micrometastases in control group (20/20).Difference was statistically significant (P =0.000).In HE-positive lymph node group:the number of CD83-positive DC in 2 times group,3 times group,and 4 times group when compared with no chemotherapy group were statistically significant (P20 =0.001,P30 =0.000,P40 =0.000).In HE-negative lymph node near foci group:the number of CD83-positive DC in 3 times group,and 4 times group compared with 0 group were statistically significant increased (P30 =0.001,P40 =0.001).In HE-negative lymph node far foci group:the number of 4 times group was significantly higher than 0,2 and 3 group (P40 =0.000,P42 =0.000,P43 =0.001).Conclusions Neoadjuvant chemotherapy is effective in the prevention of lymph node micrometastases in stage Ⅲ gastric carcinoma and it increases the number of mature dendritic cells in lymph nodes.

Objective To compare the efficacy and safety of modified FOLFOX4 program with docetaxel-oxaliplatin-5-fluorouracil (DOF) program as neoadjuvant chemotherapy in stage Ⅲ gastric cancer patients.Methods In 86 cases of stage Ⅲ gastric cancer patients,42 received modified FOLFOX4 chemotherapy,44 cases were treated by DOF program.After two cycles of chemotherapy,the treatment effect and adverse reactions were evaluated.Results The total effectiveness in modified FOLFOX4 group and DOF group was 60％ (25/42) and 68％ (30/44) respectively (P ＞ 0.05).Tumor control rate was 83％ (35/42) and 89％ (39/44) respectively (P ＞ 0.05).The incidence of nausea,vomiting and leukopenia was higher in DOF group than modified FOLFOX4 group (P ＜ 0.05).The D2 lymph node dissection rate between modified FOLFOX4 group and DOF group was 80％ (20/25) and 87％ (26/30) respectively (P ＞ 0.05),the R0 resection rate was 72％ (18/25) and 83％ (25/30) respectively (P ＞ 0.05).In 43 R0 resection patients using FOLFOX4 the median survival time was 38.7 months,1-,3-year survival rate was 90％,and 60％ ; while that in DOF group the median survival time was 39.6 months,1-,3-year survival rate was 95％,and 75％.Multivariate analysis showed that postoperative TNM stage only was an independent risk factor for prognosis.Conclusions TNM stage was the independent risk factor for prognosis of gastric carcinoma patients after radical gastrectomy.

Objective To evaluate efficacy and safety of intravenous chemotherapy combined with TACE in the treatment of gastric cancer with hepatic metastasis.Methods In this study 60 gastric cancer patients with hepatic metastasis were divided into two groups randomly,the trial group (30 cases) took intravenous chemotherapy consisted of docetaxel 60 mg/m2 intravenous drip d1,oxaliplatin 80 mg/m2 intravenous drip d1 followed by 5-fiuorouracil 500 mg/m2 intravenous drip d1-5 after TACE.The control group (30 cases) took the same chemotherapy protocol and one treatment cycle covering 4 weeks.Results The overall response rate in trial group was 53％ and the increase rate of Karnofsky was 70％ while that was 37％ and 40％ respectively in the control group (P ＜ 0.05).The median overall survival of trial group was 13 months and which was 8 months in the control group (P ＜ 0.05),meanwhile the toxicities and side reactions of trial group didn't increase notably.Conclusions Intravenous chemotherapy combined with TACE is safe and more effective in the treatment of gastric cancer with hepatic metastasis.