Objective:To study the correlation of tumor-associated macrophages infiltration , MMP-2 expression and angiogenesis in colon carcinoma by testing the MMP-2 through immunological methods ,and counting of macrophages and distributing of vessels in colonic adenocarcinoma at different differential degrees .Methods: Analyze the infiltration of tumor-associated macrophages (TAMs),and count blood capillaries in 79 patients with colon carcinoma by immunohistochemical staining ,detect the expression of MMP-2 in macrophages by immunological methods .Analyze the relationship among numbers of macrophages and vessels , clinical pathology and expression of MMP-2.Results: Macrophages in colon carcinoma were labeled to brown yellow by murine monoclonal antibody to human.The number of macrophages was markedly higher than normal group (F=412.04,P<0.05); Variance of macrophages number had statistical significance in differentiatial degrees and Duck ′s staging of colon carcinoma ( t=10.80 and F=412.04,P<0.05);variance of microvessel density(MVD) had statistical significance in Duck′s staging of colon carcinoma(t=7.35, P<0.05 ) ,MVD in colon carcinoma patients with lymphatic metastasis was higher than patients without lymphatic metastasis ( t=6.77 , P<0.05).Expression of MMP-2 in colon carcinoma showed strong positive.Expression of MMP-2 in patients with lymphatic metastasis was higher than patients without lymphatic metastasis ( t=10.91 , P<0.05 ) . There was a positive relationship among MMP-2 expression,number of tumor-associated macrophages(TAMs) and MVD(r=0.451,0.672,P<0.05,respectively).Conclusion:MMP-2 and TAMs correlated closely with the development of colon carcinoma .TAMs in colon carcinoma can promote the angiogenesis ,growth and metastasis of the tumor by up-regulating the expression of MMP-2.

Objective To observe the effect of brain-derived microvesicles (BDMVs) on cytoskeleton in human umbilical vein endothelial cells (HUVECs).Methods BDMVs were prepared in vitro and identified by transmission electron microscopy and particle size identification.HUVECs were co-cultured with PKH26-1abeled BDMVs for 0.5,1,and 2 h;flow cytometry was used to detect the phagocytosis of HUVECs for BDMVs at different time points.HUVECs cultured in vitro were divided into control group,BDMVs treatment group and nimodipine treatment group;cells in the BDMVs treatment group were given 1.5× 107/mL BDMVs;cells in the nimodipine treatment group were pretreated with 2 μg nimodipine (0.2 mg/mL) for 10 min,and then,given 1.5×107/mL BDMVs.After being stained with rhodamine-labeled phalloidin,the fluorescence intensity and number of stress fibers of fibroactin in HUVECs were observed by laser confocal microscopy.Results BDMVs had complete membrane structure with a diameter of 100-1000 nm under transmission electron microscopy.The proportion of cells phagocytizing BDMVs increased significantly with prolonged incubation time,enjoying significant differences (0.5h:22.7％±1.2％;1 h:52.3％±1.3％;2h:71.6％±1.9％,P＜0.05).Laser confocal microscopy showed that,as compared with the control group,the fluorescence intensity ofcytoskeletal protein was obviously increased and the number of stress fibers increased was obviously larger in the BDMVs treatment group.As compared with those in the BDMVs treatment group,the fluorescence intensity of cytoskeletal protein was decreased and the number of stress fibers was obviously smaller in the nimodipine group.Conclusion The role of BDMVs in phagocytosis of HUVECs becomes stronger as time being prolonged,and BDMVs phagocytosis leads to cytoskeletal remodeling,which can be partially blocked by nimodipine.

Objective:To establish a new type of small intestinal organoids with injury-related regenerative capacity, and to simulate the process of intestinal injury, regeneration, and repair in vitro. Methods:The crypt structures of ileal mucosa from 6 to 8 weeks old, 18 to 24 g specific pathogen-free C57BL/6 mice were isolated. The ENR, ENR+ tumor necrosis factor-α(TNF-α) and 8C culture systems were designed to establish small intestinal organoids under conditions of intestinal homeostasis, inflammatory injury and injury-related regeneration, and the morphology of intestinal organoids were observed. The cell types and spatial arrangements of intestinal organoids, and the expression of genes clusterin( Clu), annexin A1( Anxa1), stem cell antigen-1( Sca1) and regenerating islet-derived protein 3-beta( Reg3 b) at protein levels were detected by immunofluorescence staining. The expression of genes Clu, Anxa1, Sca1 and Reg3 b at mRNA levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Independent sample- t test was used for statistical analysis. Results:In ENR and 8C culture system, both intestinal organoids contained intestinal stem cells, goblet cells, Paneth cells and intestinal endocrine cells, and the spatial arrangement of cells was similar to the intestinal epithelium. In the 8C culture system, the amplification capacity of the new small intestinal organoids was significantly enhanced, the growth rate was faster, and the structure was larger and more complex than those of small intestinal organoids in ENR and ENR+ TNF-α culture systems. The results of qRT-PCR showed that, the relative mRNA expression levels of novel small intestinal organoid regeneration genes Clu, Anxa1, and Sca1 in the 8C culture system were higher than those in the ENR and ENR+ TNF-α culture systems (0.68±0.31 vs.0.20±0.07 and 0.36±0.19, 0.48±0.13 vs. 0.07±0.02 and 0.18±0.11, 0.56±0.20 vs. 0.02±0.01 and 0.08±0.04), and the differences were statistically significant ( t=4.82 and 2.77, 8.62 and 4.89, and 8.58 and 7.50; all P<0.05). The results of immunofluorescence staining indicated that, the expression levels of novel small intestinal organoid regeneration genes Clu, Anxa1, Sca1 and Reg3 b at protein level in the 8C culture system were higher than those in the ENR and ENR+ TNF-α culture systems (31.62±1.69 vs. 9.73±2.39 and 15.11±2.16, 42.65±1.85 vs. 19.70±1.18 and 24.97±2.82, 63.80±2.73 vs. 37.10±1.59 and 43.27±2.53, 53.26±1.84 vs. 27.75±3.78 and 33.16±3.50), and the differences were statistically significant( t=12.95 and 10.41, 18.13 and 9.09, 14.63 and 9.56, and 10.51 and 8.80; all P<0.001). Conclusion:The small intestinal organoids established in the novel culture system have the characteristics of injury-related regeneration, and provide a novel in vitro model for studying the regeneration of epithelial tissues and organs.

Objective: To explore the predictive value of the timed "up and go" test on falling of inpatients with stroke, providing objective reference for clinical fall risk evaluation. Methods: A total of 62 stroke in patients who were treated in the department of Neurology of the Second Affiliated Hospital of Shantou University Medical College from January 2018 to June 2018 were selected by the convenience sampling method. The timed "up and go" test was used to record the walking function of stroke inpatients. An ROC curve was used to analyze the efficacy in predicting falls of stroke inpatients. Results: The TUGT has demonstrated intraclass correlation coefficient (ICC) value of 0.963 (95%CI 0.938-0.978), and Cronbach alpha coefficient value of 0.983.The difference of the TUGT time between recurrent-fall stroke inpatients [15.69 (13.30, 19.87) seconds] and the patients without a history of falling [12.45 (10.08, 16.26) seconds] was statistically significant (Z=-2.585, P<0.05). Meanwhile, the difference of the TUGT time between male stroke inpatients [13.12 (10.23, 15.35) seconds] and female stroke inpatients [17.09 (12.98, 19.87) seconds] was also statistically significant (Z=-2.297, P<0.05). The TUGT time was significantly (P<0.05) positively correlated both with the fall history (rs=0.331) and gender (rs=0.294) in stroke inpatients. The result of ROC curve showed that the optimal TUGT cut-off point for predicting the fall of male stroke patients was 13.12 seconds with 0.696 for AUC, 78.6% for sensitivity and 68.2% for specificity (P<0.05).While the optimal TUGT cut-off point for predicting the fall of female stroke inpatients was 13.68 seconds with 0.691 for AUC, 73.3% for sensitivity and 65.6% for specificity (P<0.05). Conclusions Although TUGT has good feasibility in stroke inpatients, its predictive value for future falls of stroke inpatients is medium. The assessment and prediction of fall risk should be considered carefully by medical workers so as to reduce the occurrence of falls in stroke inpatients.

Objective: To investigate the differential expression of serum-and-glucocorticoid-inducible-kinase-2 (SGK2) in hepatocellular carcinoma (HCC) and normal liver tissues and the related mechanism mediating signal transduction of GSK-3 β / β catenin in HCC cells. Methods: Twenty pairs of matched HCC and normal tissues were collected and the situation of expression of SGK2 mRNA was detected by real-time fluorescence quantitative PCR. Western blot was used to detect the levels of SGK2 protein in human HCC cell lines (Huh-7, SMMC-7721) and normal human liver cell line (L02). SGK2 siRNA was used to transfect human HCC cell lines (SMMC-7721 and Huh-7), and then the protein expression levels of GSK-3 β/ β - catenin was successfully detected with the above-mentioned transfected cell line by western blot. Measurement data were expressed as mean ± standard deviation (±s), and the Student t -test was used as the statistical method. Results: SGK2 mRNA expression was up-regulated in all 20 HCC samples than that of the expression of matched normal liver tissues. SGK2 protein levels were significantly higher in Huh-7 and SMMC-7721 than normal human liver cell lines (P < 0.01). The downregulation of SGK2 expression in human HCC cell lines (SMMC-7721 and Huh-7) had inhibited the expression of unphosphorylated GSK-3 β. In addition, the downregulation of SGK2 expression in HCC cell lines had decreased the dephosphorylation of β - catenin to prevent degradation of the β - catenin proteasome. Conclusion SGK2 is overexpressed in HCC and mediates GSK-3β/β- catenin signaling in HCC cells.