Objective To investigate the effect of macrophage polarization induced by colorectal cancer exosomes on the anti-tumor activity of CD8+T cells.Methods M0 macrophages were co-incubated with PBS,HT-29 cells,and LoVo exosomes(HT-29 and LoVo exo)for 48 h,and termed the PBS,HT-29 exo,and LoVo exo groups.Real-time quantitative polymerase chain reaction was performed to detect M2 macrophage biomarker CD206,Arginase-1,IL-10,and CD163 mRNA expressions as well as M1 macrophage biomarker iNOS and IL-1β mRNA expressions.CD8+T-cells were co-incubated with the macrophages of the aforementioned groups(PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups)for 48 h.Flow cytometry was performed to detect CD8+T PD-1 expression.Next,the PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups of CD8+T-cells were incubated with HT-29 or LoVo cells for 24 h,respectively(PBS+M0/CD8+T,HT-29 exo+M0/CD8+T,and LoVo exo+M0/CD8+T groups).Subsequently,the enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of IFN-γ,perforin,and granzyme B in the cell supernatant.The cell lysis rates of HT-29 and LoVo cells were detected through cytotoxicity experiments.Results Compared with the PBS group,CD206,Arginase-1,IL-10,and CD 163 mRNA expressions of macrophages in the HT-29 exo and LoVo exo groups were significantly upregulated(P＜0.05),whereas iNOS and IL-1βmRNA expressions were significantly downregulated(P＜0.05).Compared with the PBS+M0 group,the HT-29 exo+M0 and LoVo exo+M0 groups exhibited significantly increased PD-1 expression(P＜0.05).Compared with the PBS+M0/CD8+T group,the IFN-γ,perforin,and granzyme B levels in the cell culture supernatant of the HT-29 exo+M0/CD8+T and LoVo exo+M0/CD8+T groups were significantly reduced(P＜0.05),and the cell lysis rates of HT-29 and LoVo were significantly reduced(P＜0.001).Conclusion M2 macrophage induced by HT-29 and LoVo exo can inhibit the tumor-killing function of CD8+T cells.

Objective To investigate the effect of macrophage polarization induced by colorectal cancer exosomes on the anti-tumor activity of CD8+T cells.Methods M0 macrophages were co-incubated with PBS,HT-29 cells,and LoVo exosomes(HT-29 and LoVo exo)for 48 h,and termed the PBS,HT-29 exo,and LoVo exo groups.Real-time quantitative polymerase chain reaction was performed to detect M2 macrophage biomarker CD206,Arginase-1,IL-10,and CD163 mRNA expressions as well as M1 macrophage biomarker iNOS and IL-1β mRNA expressions.CD8+T-cells were co-incubated with the macrophages of the aforementioned groups(PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups)for 48 h.Flow cytometry was performed to detect CD8+T PD-1 expression.Next,the PBS+M0,HT-29 exo+M0,and LoVo exo+M0 groups of CD8+T-cells were incubated with HT-29 or LoVo cells for 24 h,respectively(PBS+M0/CD8+T,HT-29 exo+M0/CD8+T,and LoVo exo+M0/CD8+T groups).Subsequently,the enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of IFN-γ,perforin,and granzyme B in the cell supernatant.The cell lysis rates of HT-29 and LoVo cells were detected through cytotoxicity experiments.Results Compared with the PBS group,CD206,Arginase-1,IL-10,and CD 163 mRNA expressions of macrophages in the HT-29 exo and LoVo exo groups were significantly upregulated(P＜0.05),whereas iNOS and IL-1βmRNA expressions were significantly downregulated(P＜0.05).Compared with the PBS+M0 group,the HT-29 exo+M0 and LoVo exo+M0 groups exhibited significantly increased PD-1 expression(P＜0.05).Compared with the PBS+M0/CD8+T group,the IFN-γ,perforin,and granzyme B levels in the cell culture supernatant of the HT-29 exo+M0/CD8+T and LoVo exo+M0/CD8+T groups were significantly reduced(P＜0.05),and the cell lysis rates of HT-29 and LoVo were significantly reduced(P＜0.001).Conclusion M2 macrophage induced by HT-29 and LoVo exo can inhibit the tumor-killing function of CD8+T cells.

Objective To explore the mechanism underlying ITGB1-induced drug resistance in gastric cancer based on the circRNA-miRNA-ITGB1 regulatory network.Methods Tumor tissue samples were collected from 21 patients with gastric cancer.The ITGB1 gene expression levels were determined using real-time fluorescent quantitative polymerase chain reaction,and circRNA sequencing was performed to compare the differences in circRNAs between patients with low and high ITGB1 expression.BGC-823 cells were trans-fected with si-circ_0027189,si-miR-455,or si-NC and divided into the si-circ_0027189,si-miR-455,or si-NC groups,respectively.The circ_0027189,miR-455,and ITGB1 expression levels in each group and the sensitivity to oxaliplatin were measured.Results The circRNA regulatory network showed that circ_0027189 regulated ITGB1 expression through miR-455.Compared to the si-NC group,the si-circ_0027189 group exhibited decreased expression levels of circ_0027189 and ITGB1,increased expression levels of miR-455,and reduced sensitivity to oxaliplatin.In contrast,the si-miR-455 group showed decreased expression levels of miR-455,increased expression levels of ITGB1,and enhanced sensitivity to oxaliplatin compared to the si-NC group.Conclusion circ_0027189 can increase ITGB1 expression levels by targeting miR-455 expression,ultimately increasing drug resistance in gastric cancer cells.

Objective:To investigate the association between shift work and the risk of lower extremity osteoarthritis.Methods:The study population came from the Dongfeng-Tongji cohort established in 2008. In September 2008, the Dongfeng Motor Company in Hubei Province was to recruit all retired workers who voluntarily participated in the survey as the research objects. During the follow-up conducted from April to October 2013, a total of 14 438 retired workers, i.e. all of the participants who underwent physical examination were investigated about demographic characteristics, lifestyles, occupation history, and lower extremity joint-related medical history, and additionally completed lower extremity joint examinations. After excluding individuals with missing data regarding lower extremity osteoarthritis, with the history of lower extremity joint trauma, or with history of rheumatoid arthritis (N=532), data from 13 906 participants was analyzed in the study. Multivariate logistic regression models were used to estimate the association between shift work and lower extremity osteoarthritis. After stratified by the duration of shift work, multivariate logistic regression models were used to analyze the relationship between the duration after leaving from shift work and lower extremity osteoarthritis.Results:Finally, a total of 13 906 retired employees included 7 560 (54.4%) females with a mean age of 64.74 (standard deviation 8.23) years old. 5 537 (39.8%) workers had ever engaged in shift work, including 2 004 (14.4%) workers with 1-9 years of shift work and 3 533 (25.4%) workers with ≥ 10 years of shift work. The prevalence of lower extremity osteoarthritis was 7.0%, while the prevalence of knee osteoarthritis and hip osteoarthritis were 6.7% and 0.7%, respectively. Compared with daytime workers, shift workers showed a 22% increase in the risk of lower extremity osteoarthritis ( OR=1.22, 95 %CI:1.06-1.40). Each 5-year increase in the duration of shift work was associated with a 4% increase in the risk of lower extremity osteoarthritis ( OR=1.04, 95 %CI:1.01-1.08). With the extension of the duration after leaving from shift work, the risk of lower extremity osteoarthritis decreased. Similar relationships were found between shift work and the risk of knee osteoarthritis, as well as hip osteoarthritis. Conclusion:Shift work was associated with the increased risk of lower extremity osteoarthritis.

Objective:To investigate the association between shift work and the risk of lower extremity osteoarthritis.Methods:The study population came from the Dongfeng-Tongji cohort established in 2008. In September 2008, the Dongfeng Motor Company in Hubei Province was to recruit all retired workers who voluntarily participated in the survey as the research objects. During the follow-up conducted from April to October 2013, a total of 14 438 retired workers, i.e. all of the participants who underwent physical examination were investigated about demographic characteristics, lifestyles, occupation history, and lower extremity joint-related medical history, and additionally completed lower extremity joint examinations. After excluding individuals with missing data regarding lower extremity osteoarthritis, with the history of lower extremity joint trauma, or with history of rheumatoid arthritis (N=532), data from 13 906 participants was analyzed in the study. Multivariate logistic regression models were used to estimate the association between shift work and lower extremity osteoarthritis. After stratified by the duration of shift work, multivariate logistic regression models were used to analyze the relationship between the duration after leaving from shift work and lower extremity osteoarthritis.Results:Finally, a total of 13 906 retired employees included 7 560 (54.4%) females with a mean age of 64.74 (standard deviation 8.23) years old. 5 537 (39.8%) workers had ever engaged in shift work, including 2 004 (14.4%) workers with 1-9 years of shift work and 3 533 (25.4%) workers with ≥ 10 years of shift work. The prevalence of lower extremity osteoarthritis was 7.0%, while the prevalence of knee osteoarthritis and hip osteoarthritis were 6.7% and 0.7%, respectively. Compared with daytime workers, shift workers showed a 22% increase in the risk of lower extremity osteoarthritis ( OR=1.22, 95 %CI:1.06-1.40). Each 5-year increase in the duration of shift work was associated with a 4% increase in the risk of lower extremity osteoarthritis ( OR=1.04, 95 %CI:1.01-1.08). With the extension of the duration after leaving from shift work, the risk of lower extremity osteoarthritis decreased. Similar relationships were found between shift work and the risk of knee osteoarthritis, as well as hip osteoarthritis. Conclusion:Shift work was associated with the increased risk of lower extremity osteoarthritis.

Objective_To evaluate the proangiogenesis of extracelluar matrix proteins SRPX2 on HUVECs.Meth-ods_pcDNA3.1-SRPX2 vector ( transfection group) and pcDNA3.1 vector ( negative group) were transfected into HEK293T cells, then divided into 3 groups including blank group.The proliferation of HUVECs ( absorbance, A450 ) was detected by CCK-8 kit.The transmembrane cell number was counted by Transwell migration and wound healing assay to evaluate the migration ability of HUVECs.A three dimensional culture system of cells was construc-ted on the Matrigel, and tube formation number of HUVECs was assessed.Results_The proliferation of HUVECs ( absorbance,A450 ) among transfection group,negative group and blank group failed to show significant difference (P>0.05).A signifiant difference was noted in the total branch point of capillary tubes among transfection group, negative group and blank group[(97 ±4)/field versus (57 ±3) and (54 ±3)/field] (P<0.05).In wound healing
assay, the distance of transfection group compared to 0, 6 and 12 h were both significantly larger than that of nega-tive group and blank group [(90 ±6), (37 ±7),(36 ±4)μm and (135 ±5),(65 ±8),(63 ±4)μm respective-ly, both( P<0.05) ] .In Tranwell assay, the number of migrating cells in the transfection group was significantly more than negative group and blank group [(549 ±10)/field vs (334 ±11) and (329 ±12)/field(P<0.05)] af-ter co-culture 16 h.Conclusions_SRPX2 may promote angiogenesis by stimulating the migration and tubing on the Matrigel of HUVECs.