AIM: To assess the expression of Ki67 as prognosticator in rectal/recto sigmoid cancer.METHODS: Samples from 146 patients with rectal and recto sigmoid cancer were studied for expression of Ki67 and its prognostic sig...AIM: To assess the expression of Ki67 as prognosticator in rectal/recto sigmoid cancer.METHODS: Samples from 146 patients with rectal and recto sigmoid cancer were studied for expression of Ki67 and its prognostic significance in comparison with clinicopathological predictors of survival. Formalin-fixed, paraffin-embedded tissues from 6 (4.1%) patients with T1, 26 (17.8%) with T2, 94 (64.4%) with T3, and 20 (13.7%) with T4 tumors were studied. Ki67 expression was determined immunohistochemically. Samples were divided according to mean value into high (＞40%) and low (≤40%) expression. Areas of extensive proliferation (＞50%) were defined as 'hot spot' areas. RESULTS: Hot spot areas were present in samples regardless of histopathological grade. Lower TNM and Dukes stage and higher expression of Ki67 and presence of Ki67 hot spot areas in histopathological samples were associated with better survival, whereas no association was observed with histopathological grade (P = 0.78). In Cox multivariate regression analysis, significant prognostic factors were Dukes stage (P＜0.001), presence of lymph node metastases (P = 0.015), age (P = 0.035) andpresence of Ki67 hot spot areas (P = 0.044). CONCLUSION: Proliferative activity as measured by Ki67 in rectal cancer is associated with survival improvement compared with patients with low Ki67. Areas of prognostically significant increased proliferation were found independently of histopathological tumor grade.展开更多
The effect of monoethylphosphate (MEP, commercial available or synthesized) together with IL-2 on the selective proliferation of human γ~δ T cells in Vitro from peripheral blood mononuclear cells (PBMC) of healthy d...The effect of monoethylphosphate (MEP, commercial available or synthesized) together with IL-2 on the selective proliferation of human γ~δ T cells in Vitro from peripheral blood mononuclear cells (PBMC) of healthy donors and of cancer patients was investigated. The γ~δ T cells were stimulated by MEP to proliferate in a dose-dependent manner. The effect of synthesized MEP was 10 times greater than that of commercial MEP. When the PBMCs of healthy donors were cultured for 25 d in the medium containing different concentrations of MEP, the total cell number increased about 1000-3000 fold; and the ratio of γ~δ T cells reached to 70-80%. The selective expansion of γ~δ T cells depended on the synergic action of MEP and IL-2. The bulk cultured γ~δ T cells exhibited obvious cytotoxic activities against allogenic tumor cell lines (SQ-5,K562 alld Daudi) and autologous tumor cells. The culture system described here not only offers a simple method for obtaining a large number of γ~δ T cells which may become a new effector in the adoptive immunotherapy, but also provides a useful model for the further studies of the structure and function of γ~δ T cells in vitro.展开更多
Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral p...Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral proteins. To achieve this goal, we constructed a DNA vaccine expressing nonstructural 3 (NS3) gene (pcDNA3.1-HCV-NS3) and assessed the immune response in C57BL/6 mice. In this study, the NS3 gene was amplified with a nested-reverse transcriptase-polymerase chain reaction (RT-PCR) method using sera of HCV-infected patients with genotype 1a. The resulting NS3 gene was subcloned into a pcDNA3.1 eukaryotic expression vector, and gene expression was detected by western blot. The resultant DNA vaccine was co-administered with interleukin-12 (IL-12) as an adjuvant to female C57BL/6 mice. After the final immunizations, lymphocyte proliferation, cytotoxicity, and cytokine levels were assessed to measure immune responses. Our data suggest that co-administration of HCV NS3 DNA vaccine with IL-12 induces production of significant levels of both IL-4 and interferon (IFN)-γ (p<0.05). Cytotoxicity and lymphocyte proliferation responses of vaccinated mice were significantly increased compared to control (p<0.05). Collectively, our results demonstrated that co-administration of HCV NS3 and IL-12 displayed strong immunogenicity in a murine model.展开更多
Objective: To detect the different morphologic features, developmental regulation, potential of proliferation and differentiation of neonatal rat cochlea progenitor spheres. Methods: We isolated the cochlea sensory ep...Objective: To detect the different morphologic features, developmental regulation, potential of proliferation and differentiation of neonatal rat cochlea progenitor spheres. Methods: We isolated the cochlea sensory epithelium cells from neonatal rats and cultured them in nonadherent conditions to acquire different morphologic spheres. Then we observed the diameter and compositional change of cell colonies in distinct sphere types on day 3, 6, 9 and 12, and summarized the regularity of development and their conversion. We also detected the proliferative activity of distinct spheres by immunohistochemical staining of Abcg2, Nestin and BrdU. After induced spontaneous differentiation, the spheres were detected in the changes of the marker of hair cell, MyosinVIIA; by immunocytochemical staining, we revealed the potential of how different spheres were converted into hair cell-like cells. Results: The acquired three types of suspended spheres are solid, transitional, and hollow. There's morphologic significance among them and they can covert into the other type of spheres among them. The ability of self-renewing and proliferation in distinct spheres vary and all of them have the potential of spontaneously differentiation into hair cell-like cells. Conclusion: All the type of spheres not only has the potential of proliferation and differentiation, but also hasthe potential of spontaneous differentiation into hair cell-like cells. Distinct types of cell spheres neither originate from different progenitor cell subcolonies nor are different stages of the same cell spheres. Solid spheres are most practically useful.展开更多
基金Supported by the Emil Aaltonen Foundation and Turku University Research Foundation
文摘AIM: To assess the expression of Ki67 as prognosticator in rectal/recto sigmoid cancer.METHODS: Samples from 146 patients with rectal and recto sigmoid cancer were studied for expression of Ki67 and its prognostic significance in comparison with clinicopathological predictors of survival. Formalin-fixed, paraffin-embedded tissues from 6 (4.1%) patients with T1, 26 (17.8%) with T2, 94 (64.4%) with T3, and 20 (13.7%) with T4 tumors were studied. Ki67 expression was determined immunohistochemically. Samples were divided according to mean value into high (＞40%) and low (≤40%) expression. Areas of extensive proliferation (＞50%) were defined as 'hot spot' areas. RESULTS: Hot spot areas were present in samples regardless of histopathological grade. Lower TNM and Dukes stage and higher expression of Ki67 and presence of Ki67 hot spot areas in histopathological samples were associated with better survival, whereas no association was observed with histopathological grade (P = 0.78). In Cox multivariate regression analysis, significant prognostic factors were Dukes stage (P＜0.001), presence of lymph node metastases (P = 0.015), age (P = 0.035) andpresence of Ki67 hot spot areas (P = 0.044). CONCLUSION: Proliferative activity as measured by Ki67 in rectal cancer is associated with survival improvement compared with patients with low Ki67. Areas of prognostically significant increased proliferation were found independently of histopathological tumor grade.
文摘The effect of monoethylphosphate (MEP, commercial available or synthesized) together with IL-2 on the selective proliferation of human γ~δ T cells in Vitro from peripheral blood mononuclear cells (PBMC) of healthy donors and of cancer patients was investigated. The γ~δ T cells were stimulated by MEP to proliferate in a dose-dependent manner. The effect of synthesized MEP was 10 times greater than that of commercial MEP. When the PBMCs of healthy donors were cultured for 25 d in the medium containing different concentrations of MEP, the total cell number increased about 1000-3000 fold; and the ratio of γ~δ T cells reached to 70-80%. The selective expansion of γ~δ T cells depended on the synergic action of MEP and IL-2. The bulk cultured γ~δ T cells exhibited obvious cytotoxic activities against allogenic tumor cell lines (SQ-5,K562 alld Daudi) and autologous tumor cells. The culture system described here not only offers a simple method for obtaining a large number of γ~δ T cells which may become a new effector in the adoptive immunotherapy, but also provides a useful model for the further studies of the structure and function of γ~δ T cells in vitro.
文摘Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral proteins. To achieve this goal, we constructed a DNA vaccine expressing nonstructural 3 (NS3) gene (pcDNA3.1-HCV-NS3) and assessed the immune response in C57BL/6 mice. In this study, the NS3 gene was amplified with a nested-reverse transcriptase-polymerase chain reaction (RT-PCR) method using sera of HCV-infected patients with genotype 1a. The resulting NS3 gene was subcloned into a pcDNA3.1 eukaryotic expression vector, and gene expression was detected by western blot. The resultant DNA vaccine was co-administered with interleukin-12 (IL-12) as an adjuvant to female C57BL/6 mice. After the final immunizations, lymphocyte proliferation, cytotoxicity, and cytokine levels were assessed to measure immune responses. Our data suggest that co-administration of HCV NS3 DNA vaccine with IL-12 induces production of significant levels of both IL-4 and interferon (IFN)-γ (p<0.05). Cytotoxicity and lymphocyte proliferation responses of vaccinated mice were significantly increased compared to control (p<0.05). Collectively, our results demonstrated that co-administration of HCV NS3 and IL-12 displayed strong immunogenicity in a murine model.
基金Supported by the National Natural Science Foundation of China (Grant No.30973300)
文摘Objective: To detect the different morphologic features, developmental regulation, potential of proliferation and differentiation of neonatal rat cochlea progenitor spheres. Methods: We isolated the cochlea sensory epithelium cells from neonatal rats and cultured them in nonadherent conditions to acquire different morphologic spheres. Then we observed the diameter and compositional change of cell colonies in distinct sphere types on day 3, 6, 9 and 12, and summarized the regularity of development and their conversion. We also detected the proliferative activity of distinct spheres by immunohistochemical staining of Abcg2, Nestin and BrdU. After induced spontaneous differentiation, the spheres were detected in the changes of the marker of hair cell, MyosinVIIA; by immunocytochemical staining, we revealed the potential of how different spheres were converted into hair cell-like cells. Results: The acquired three types of suspended spheres are solid, transitional, and hollow. There's morphologic significance among them and they can covert into the other type of spheres among them. The ability of self-renewing and proliferation in distinct spheres vary and all of them have the potential of spontaneously differentiation into hair cell-like cells. Conclusion: All the type of spheres not only has the potential of proliferation and differentiation, but also hasthe potential of spontaneous differentiation into hair cell-like cells. Distinct types of cell spheres neither originate from different progenitor cell subcolonies nor are different stages of the same cell spheres. Solid spheres are most practically useful.
