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Objective To investigate the value of leucine-rich repeat-containing G protein coupled receptor 5 (LGR5) and aldehyde dehydrogenase 1A1 (ALDH1A1) in progression and prognosis of non small-cell lung cancer (NSCLC),in order to find new targets for NSCLC-targeted imaging and radiation therapy.Methods Fresh tissues (n =24) and paraffin embedding tissues (n =109) of patients with NSCLC were collected from the First Affiliated Hospital of Soochow University between November 2009 and March 2012.Quantitative real time-PCR was used for the investigation of expression of LGR5 and ALDH1A1 mRNA in 24 NSCLC patients.Immunohistochemistry (IHC) was used for detecting LGR5 and ALDH1A1 expressions in NSCLC tissues and adjacent normal tissues.Data were analyzed by Mann-Whitney u test,x2 test,Pearson correlation analysis,Kaplan-Meier method,Cox proportional hazards regression model.Results Compared with adjacent normal tissues,LGR5 and ALDH1A1 mRNA were frequently increased in NSCLC tissues (u values:150,74,both P＜0.01),and the expression of LGR5 and ALDH1A1 mRNA was significantly correlated (r=0.416,P＜0.05).Positive LGR5 and ALDH1A1 expression was defined in 28.44％ (31/109) and 41.28％ (45/109) of the NSCLC tumors,respectively.Further analysis indicated that 24 of the LGR5 positive samples (77.42％,24/31) expressed ALDH1A1 (r=0.388,P＜0.01).LGR5 and ALDH1A1 ex pressions in NSCLC with higher TNM stage were significantly higher than those in NSCLC with lower TNM stage (x2 values:4.64,5.24,both P＜0.05).Coexpression of LGR5 and ALDH1A1 in NSCLC with lymph node metastasis was higher than that in NSCLC without lymph node metastasis (x2=4.12,P＜0.05).High expression of LGR5 or ALDH1A1 was related to poor prognosis (x2 values:6.24,4.18,both P＜0.05),and NSCLC patients with coexpression of LGR5 and ALDH1A1 had a poorer prognosis than the others (x2 =10.63,P＜0.01).Both of them were independent risk factors of a poorer prognosis (corrected hazard ratio (95％ CI):2.361(1.106-5.037),2.306(1.101-4.830);both P＜0.05).Conclusions The expressions of LGR5 and ALDH1A1 are closely associated with the tumorigenesis,metastasis and poor prognosis of NSCLC.LGR5 and ALDH1A1 might be new targets for NSCLC-targeted tumor imaging and radiation therapy.
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Objective To explore the expression of peripheral T cell subgroup (CD3+,CD4+,CD8+,CD4+CD25 high regulatory T cells) and the level and significance of serum cytokines in patients with silicosis.Methods One hundred and six cases patients with silicosis were collected in the Fifth People''s Hospital of Suzhou as study subjects and 56 healthy subjects as control group.Flow cytometry was used to detect the peripheral CD3+,CD4+,CD8+ and CD4+CD25 high regulatory T cells (Treg) of the patients and the control group,while chemiluminescence immunoassay was utilized to measure the peripheral serum soluble interleukin-2 receptor (sIL-2R),interleukin 6 (IL-6),interleukin 8 (IL-8) and tumor necrosis factor α (TNF-α).Results (1) The percentages of peripheral CD3+,CD4+ and CD8+ in the silicosis group were all lower than those in the control group (t=3.755,3.828,2.347,P<0.05);the percentage of Treg cells was higher in the silicosis group than in the control group,the difference was statistically significant (t=-8.345,P<0.05).Compared with the control group,based on the one-way analysis of variance,the differences in CD3+,CD4+,CD8+ and CD4+CD25 high cells were all statistically significant (F=5.620,8.007,26.71,P<0.05);in the silicosis group,the percentage of CD4+ T cells was lower in stage III than in stage I (t=3.424,P<0.05);compared with the control group,the percentages of Treg cells in the silicosis group were lower in all stages (t=-7.934,-9.445,-5.096,P<0.05).(2) The levels of peripheral sIL-2R,IL-6,IL-8 and TNF-α in the silicosis group were higher than those in the control group,the difference were statistically significant(t=-6.952,-4.506,-2.551,-5.670,P<0.05);compared with the control group,based on the one-way analysis of variance,the differences in sIL-2R,IL-6 and TNF-α in all stages were statistically significant (F=11.03,11.31,13.22,P<0.0001);the sIL-2R was higher in patients with stage III silicosis than that of stage I (t=-2.882,P<0.05);IL-6 was significantly higher in stage II and III silicosis group than that of stage I group (t=-3.022,-2.632,P<0.05),and TNF-α was higher in patients with stage II silicosis than patients with stage I silicosis (t=-2.322,P<0.05).(3) The level of peripheral Treg cells was negatively correlated with the percentages of CD3+ and CD8+ cells in patients with silicosis (r=-0.357,-0.508,P<0.05);sIL-R2 was positively correlated with IL-6,IL-8 and TNF-α,respectively (r=0.483,0.199,0.392,P<0.05);TNF-α was positively correlated with IL-6 and IL-8,respectively (r=0.338,0.338,P<0.05).Conclusion Patients with silicosis have abnormal expression in peripheral T cell subgroups,significantly increased Treg cell and dysfunctional cytokines,which may be associated with the pathogenesis of silicosis,the detection of these indicators may have significance of diagnosis,staging,disease monitoring and prognosis of the diseases.
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Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.