Objective: To investigate the expression and prognostic significance of insulin-like growth factor binding protein 5 (IGFBP5) in gastric adenocarcinoma (GAC). Methods:IGFBP5 expression in tissue samples from 236 GAC patients was analyzed us-ing immunohistochemical methods. These patients had undergone surgical resection between 20003 and 2006 in Sun Yat-Sen Universi-ty Cancer Center. The relationship between IGFBP5 expression and clinicopathological factors in the 236 GAC patients was analyzed using Pearson correlation analysis. The significance of IGFBP5 in predicting the survival status of these patients was analyzed using Ka-plan-Meier survival analysis and Cox's proportional hazards regression model. Results:Immunohistochemical staining data indicated that IGFBP5 expression was significantly decreased in 159 of the total GAC cases (67.4%). Of the 62 cases with well-and moderately differentiated GAC, 31 (50%) exhibited reduced IGFBP5 expression. Of the 174 cases with poorly differentiated GAC, 128 showed re-duced IGFBP5 expression. Reduced IGFBP5 expression was also observed in female patients and in patients with tumors over 5 cm in size or with poorly differentiated tumors (P<0.05). The reduced expression of IGFBP5 was common in the tumors that were staged as T3+4a/b andⅢ/Ⅳ(P<0.05). Kaplan-Meier survival analysis revealed that the reduced expression of IGFBP5 was associated with poor prognosis in GAC patients (P<0.001). Cox regression analysis identified IGFBP5 expression as an independent prognostic factor for the overall survival of these cancer patients (HR=1.897, P=0.029). Conclusion: IGFBP5 expression is reduced in GAC tissues, and IGFBP5 independently predicts an unfavorable prognosis in GAC patients.

Objective:To investigate the comfort of postoperative rectal cancer patients with indwelling anal tube and analyze the influencing factors, so as to provide a basis for the formulation of prevention and improvement measures.Methods:The subjective comfort self-report method of digital score and visual analog score was used to investigate and analyze 64 patients with indwelling anal canal in anorectal surgery.Results:The main discomfort of patients with indwelling anal tube were difficulty in sitting and getting in and out of bed, of which 43 cases (61.9%) reported moderate and severe sitting difficulty, and 30 cases (46.88%) reported moderate and severe difficulty in getting up and down the bed. The exposed length of anal canal was the influencing factor of discomfort ( Fvalues were 22.018, 18.213, P<0.01). Conclusion:Rectal cancer patients with indwelling anal tube have serious difficulties in sitting position and getting in and out of bed. Corresponding measures should be taken according to the influencing factors and effective intervention methods should be used to improve the comfort of the patients.

Objective To study the role of endothelial cells on the inflammatory cytokine release in septic shock through the septic shock serum stimulating human primary endothelial cells (HPAEC) and peripheral blood mononuclear cells(PBMC).Methods PBMC isolated from healthy people by density gradient centrifugation.HPAEC cell surface markers CD144 and von Willebrand factor(vWF) molecule expression by RT-PCR and Western blot.Serum levels of IL-6,TNF-α,MCP-1 from septic shock patients and healthy human detected by ELISA.HPAEC and PBMC were stimulated with the isolated serums and LPS,respectively.ELISA was used to detect the supernatant IL-6,TNF-α,MCP-1 levels.HPAEC membrane molecules ICAM-1 expression was detected by flow cytometry with serum shock and LPS stimulation.Supernatant levels of IL-6,TNF-α,MCP-1 of HPAEC with S1P1 receptor agonist CYM-5442 pretreatment was detected by ELISA after shock serum stimulation.Results Endothelial cell markers CD144 and vWF molecules could be detected in the HPAEC.Levels of inflammatory cytokines IL-6,TNF-α,MCP-1 in patients with septic shock serum were significantly higher than healthy people (P＜0.01).PBMC and HPAEC with LPS or shock serum treatment respectively,compared with normal group,levels of inflammatory cytokines in the culture supernatant were significantly higher(P＜0.01).For PBMC,the level of inflammatory cytokines between shock group and LPS group were not significantly different (P＞0.05).But for HPAEC,levels of inflammatory cytokines in the supernatant of the shock group compared to the LPS group was significantly higher (P＜0.01).Similarly,when two cells after LPS stimulation,IL-6,TNF-α levels of HPAEC's supernatant were significantly lower than PBMC' s (P＜0.01),MCP-1 levels was no difference (P＞ 0.05).But when the stimulation of shock serum,HPAEC of IL-6,TNF-α levels and PBMC no significant difference (P ＞0.05).MCP-1 was significantly increased (P＜0.01).Shock patients serum stimulation S1P1 receptorspecific agonist CYM-5442 pretreatment of HPAEC with pretreatment of S1P1 receptor specific agonist CYM-5442,the culture supernatant of inflammatory cytokines IL-6,TNF-α,MCP-1 levels were significantly lower (P＜0.01).Conclusion Endothelial cells may play a central role on the release of inflammatory cytokine during septic shock.

Objective To investigate the roles of SENP1 in regulation of biological characteristics of NK cells.Methods Lentivirus-mediated-Senp1-small-hairpinRNA (shRNA) transduction was applied to NK92 cells.The expression of SENP1 in NK92 cells was determined by real-time PCR and Western blot.The proliferation of NK92 cells was detected by CCK-8 assay.The apoptosis of NK92 cells was determined by Annexin Ⅴ and PI labeling.The cytotoxicity of NK92 cells against K562 cells was evaluated by luciferase reporter assay.Results Treatment of NK92 cells with IL-21 resulted in SENP1 upregulation.Lentivirus mediated SENP1 knockdown reduced proliferation and increased apoptosis in NK-92 cells,but SENP1 inhibition had slight impact on the cytotoxic ability of NK92 cells to kill K562 cells.Conclusion SENP1 mediates the regulatory effect of IL-21 on the proliferation and survival of NK92 cells.

Objective To analyze the genetic characteristics and the evolution of the influenza A (H3N2) virus strains circulating in Jiangsu province between 2013 and 2014.Methods This study analyzed thirty-one representative strains of influenza A (H3N2) virus, which were isolated in different regions of Jiangsu province and during different time periods from 2013 to 2014.Results Genetic distances in nucleic acid and amino acid between a strain used for vaccine production (A/Texas/50/2012) and the 31 strains were 0.010 5 and 0.012 4.Similarities between them in nucleic acid and amino acid sequences were 97.9%-99.6% and 97.2%-99.3%.Phylogenetic analysis showed that the hemagglutinin (HA) genes of the 31 strains were divided into three different groups.Three strains isolated in 2013 and three strains isolated in 2014 belonged to Group 1 and Group 2, respectively, while the others belonged to Group 3.Three positive selection sites (237, 366 and 367) in HA protein were observed by REL model.Compared with the strain used for vaccine production, the 31 strains were characterized by amino acid substitutions (N128A/T and P198S/A) in HA protein and all of the mutations located in B-cell epitopes.The total number of mutation sites reached 24.Compared with the A/Texas/50/2012 strain, seven strains presented the glycosylation site 126NWT, and three strains showed disappeared glycosylation sites of 45NSS and 144NNS.Evaluation of vaccine efficacy for A(H3N2) virus strains showed that the vaccine efficacy was not very well.Conclusion The HA gene of A(H3N2) virus had undergone a greater variation and the vaccine efficacy was not very well in Jiangsu province during 2013 to 2014, which made the influenza A(H3N2) virus become the circulating strain.

For fine management of the clinical practice teaching in the rehabilitation therapy, a set of Rehabilitation Therapy Clinical Practice Examination was developed. It was used to assess the skill of rehabilitation evaluation, rehabilitation therapy and related knowledge of every internships, to avoid the arbitrary result of the practice examination, and has been well accepted in practice.

Objective:To design the small ubiquitin modification-fibroblast growth factor receptor 4 (SUMO-FGFR4) fusion gene and construct the expression vector pET22b-SUMO-FGFR4, to optimize the expression conditions. Methods:The SUMO-FGFR4 fusion gene was obtained by Overlap PCR and was connected to pET22b;the recombinant expression vector pET22b-SUMO-FGFR4 was obtained. The influence of lactose concentration, induction time,induction temperature,induction point and adding mode of lactose in the expression levels was observed,and the best induction condition was determined; then the solubility of recombinant protein was analyzed.Results:The SUMO-FGFR4 fusion protein was highly expressed,the molecular weight of the fusion protein was about 40 000 and it could bind with FGFR4 specific antibody.When the lactose concentration was 1.0 g·L-1 ,the induction time was 3 h,the induction temperature was 37℃,the value of A (600)was 0.8,the expression level was highest;but adding mode of lactose had no remarkable effect on the protein expression.The expression level of recombinant protein induced by lactose was higher than IPTG.SUMO-FGFR4 protein existed in a form of inclusion body.Conclusion:The SUMO-FGFR4 fusion protein is expressed successfully in this study while lactose is used as inducer and the best expression conditions are confirmed.

Objective:To construct GJB2 gene mutaitons common in Chinese EGFP fusion protein vectors, and to search for better way to study the mechanism of deletion mutaitons in GJB2 gene. Method: Non-fusion protein vectors of 235delC, 299-300 del AT and 176 del 16 bp were first made by point mutaiton methods in vitro. Then expression part of the upper 3 mutations were amplified by PCR and the PCR products were cloned into TA cloning vector. After cutting by restriction enzymes EcoRI/BamHI, three deletion mutaions were inserted into pEG-FP-N1 vector. Sequencing was used to verify the validity of the fusion protein vectors. HEK293 cells were trans-fected with the recombinant DNA samples by the liposome complex method. Results The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence singals were distributed uniformly in cytoplasm. Conclusion; GJB2 mutations common in Chinese EGFP fusion protein vectors were constructed successfully. It may provide a better way to explore the reasons of nonsyndromic hearing loss common in Chinese.

Objective To investigate the structural changes of gut microbiota in patients with Parkinson's disease ( PD).Methods Twenty-four PD patients and 14 healthy controls from Beijing Hospital in 2015 were recruited in this cross-sectional study.The general clinical information was collected and all subjects were assessed with Parkinson's disease related scales.The gut microbiota status between two groups was analyzed after extracting feces'DNA and carrying out high-throughput sequencing of bacterial 16S rRNA.Results At the phylum level, actinobacteria (0.76%(0.13%, 1.85%) vs 0.14%(0.07%, 0.30%), Z＝2.784, P＜0.01) were significantly increased and bacteroidetes (57.28%(48.75%, 64.95%) vs 63.78%(56.72%, 68.21%), Z＝-4.963, P＜0.01) were significantly decreased in PD patients compared to healthy controls.At the class level, bacilli (0.52%(0.11%, 2.10%) vs 0.13%(0.05%, 0.16%), Z＝2.693, P＜0.01), negativicutes (5.04%(2.93%, 14.02%) vs 2.87%(1.46%, 4.43%), Z＝2.360, P＝0.018), actinobacteria (0.60%(0.10%, 1.59%) vs 0.12%(0.04%, 0.20%), Z＝2.512, P＝0.011 ), gammaproteobacteria ( 1.72%( 0.58%, 5.46%) vs 0.43%(0.24%, 2.19%), Z＝2.179, P＝0.029) were significantly increased in PD patients compared to healthy controls.At the family level, veillonellaceae (3.78%(0.53%, 13.82%) vs 0.49%(0.08%, 3.14%), Z＝2.754, P＜0.01), streptococcaceae (0.33%(0.09%, 0.69%) vs 0.19%(0.14%, 0.24%), Z＝1.770, P＝0.004), enterobacteriaceae (1.04%(0.40%, 4.95%) vs 0.20%(0.10%, 0.45%), Z＝2.784, P＜0.01 ), lactobacillaceae ( 0.079%( 0.014%, 0.575%) vs 0.003%(0.002%, 0.028%), Z＝3.119, P＜0.01), bifidobacteriaceae (0.60%(0.09%, 1.57%) vs 0.11%(0.03%, 0.19%), Z＝2.481, P＝0.012) were significantly increased and pasteurellaceae (0.009%(0.002%, 0.047%) vs 0.110%(0.022%, 0.898%), Z＝-2.545, P＝0.010) were significantly decreased in PD patients compared to healthy controls.Conclusions The structures of gut microbiota in PD patients and healthy controls were significantly different at the levels of phylum , class, and family.All these changes are potentially associated with the development of PD pathology.