Objective:The effect of rat mesenchymal stem cells(MSCs)on graft versus host disease after allogeneic co-transplantation with bone marrow in rat was investigated.Methods:MSCs from bone marrow cells of rat were isolated and cultured in vitro.The purity of MSCs was identified with the spindle-fibroblastic morphological characterization and the phenotypes were analyzed with flow cytometry.The suppressive effect of MSCs on T cell proliferation was analyzed using mixed lymphocyte cultures(MLR).Allogeneic BMT model from SD rats to Wistar rats was established,and then,MSCs were co-transplanted with bone marrow cells at different ratio,the suppressive effect of MSCs on GVHD was studied by clinical symptom,Kaplan-Meier survival curve,and histological analysis.Results:In vitro,when MSC were added to the mixed lymphocyte cultures,the proliferation of T cells were inhibited markedly in a dose-dependent manner(P

Interleukin (IL)-27 is a new member of the IL-6/IL-12 family composed of p28 subunit and Epstein-Barr virus induced gene 3 (EBI3) subunit. Its receptor is composed of WSX-1 and gp130. It has dual properties including pro-inflammatory and anti-inflammatory function at different conditions. Studies have shown that IL-27 exerts its biological activities through stimulating JAK1/STAT1, JAK1/STAT3 signal pathways and regulating the production of Th1, Th17 as well as their related-cytokines. Furthermore, IL-27 can exert the role of anti-tumor activity by enhancing the effect of cytotoxic T cells and anti-angiogenesis.

Background and purpose:Esophageal cancer is a serious disease threatening human health, and it is very difficult to understand the development mechanism and find the therapeutic methods for esophageal cancer. In recent years, B7-H3, as a new member of B7 immunoregulatory superfamily, overexpressed in multiple tumor types, is considered to be a new tumor marker and potential therapeutic target. This study aimed to detect the expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13, Eca-109 and exploring the effect of B7-H3 siRNA on cell proliferation, migration and invasionin vitro in human esophageal cancer Eca-109 cell line. Methods:The expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13 and Eca-109 were detected by reverse transcription polymerase chain reaction (RT-PCR). B7-H3 siRNA and control siRNA were transfectedin vitro into human esophageal cancer Eca-109 cells using LipofectamineTM 2000. The expressions of B7-H3 mRNA and protein in Eca-109 cells were analyzed by RT-PCR and Western blot. The proliferation, migration and invasion abilities of Eca-109 cells were measured by MTT assay, wound scrape assay and transwell invasion assayin vitro,respectively.Results:All tested cultured esophageal cancer cell lines constitutively expressed B7-H3 mRNA under normal conditions (TE-1 0.382±0.008, TE-13 0.399±0.008, Eca-109 0.428±0.012). After transfection, the expression of B7-H3 mRNA levels decreased in B7-H3 siRNA transfected group, compared with control siRNA transfected group (0.128 5±0.000 2vs 0.532 4±0.000 7,P<0.01) and untransfected group (0.128 5±0.000 2vs 0.540 3±0.001 3,P<0.01), while its protein expression levels were also signiifcantly lower than the control transfection group (0.421 4±0.004 8vs 0.500 6±0.012 9,P<0.05) and untransfected group (0.421 4±0.004 8vs 0.492 1±0.014 8, P<0.05). Compared with control transfected and untransfected cells, Eca-109 cell migration and invasion abilities decreased significantly (P<0.05) by siRNA interference, but no significant difference was observed between their proliferative capacity (P>0.05).Conclusion:All tested esophageal cancer cell lines constitutively express B7-H3 mRNA. B7-H3 siRNA interference inhibits Eca-109 cell migration and invasion abilities. B7-H3 may have a critical role in regulating Eca-109 cell progression.

Objective:Tumor common antigen of an extract of plant and applying in early stage diagnosis for tumor were studied.Methods:Tumor cells and lymphocytes proliferate responses were determined by 3H-TdR intervening.Fresh tumor tissue from human transplant to mice,the survival rate of transplanted tumor was observed.DNA content of tumor cells was measured by flowcytometery.The chemical nature was analyzed by chromatography and measured by ultraviolet-spectrometer.Results:The extract of plant markedly stimulated the lymphocytes from tumor patients and atypical hyperplasia patient to proliferate,but failed to health persons(P

Objective:To study the expression and correlation of tumor-associated macrophages(TAM) and CCL5 in ganstric cancer.Methods:48 cases patients with completed clinical and pathological data of gastric cancer paraffin block specimens were select-ed.Cancer tissues and adjacent tissues were used as control,using SP immunohistochemical method to detect CD68 and CCL5 in gastric cancer tissues and adjacent tissues,and using the Spearman correlation statistics statistical methods for the correlation.Results:CD68 and CCL5 showed positive expression in gastric cancer tissue,significantly higher than those in the adjacent tissues(P<0.01),CD68 and CCL5 were related with gastric cancer invasion depth, lymph node metastasis, TNM stage and tumor differentiation ( P<0.001 ) . There was positive relation between the expression of CD68 and CCL5 in gastric cancer(P<0.01,r=0.759).Conclusion: CD68 and CCL5 played a driving role to the invasion and metastasis of gastric cancer occurrence,suggesting that the secretion CCL5 by TAM may promote the invasion and metastasis of gastric cancer.

Objective:To study the influence of the DCs from mononuclear cells in axillary draining lymph nodes of patients with breast cancer to the cytotoxicity of the antigenic specific CTLs stimulated by freeze-thrawing antigen from breast cancer cells in vitro.Methods:The mononuclear cells were isolated from axillary draining lymph nodes,and were cultured with cytokines to induce DCs and tumor draining lymph node cells(TDLNCs) respectively.DCs stimulated by the auto-breast cancer freeze-thrawing antigen(DCs-Ag).TDLNCs were co-cultured with DCs-Ag to derive tumor antigenic specific CTLs(DC-Ag-TDLNC).Then the cytotoxicty of specific CTLs to auto-breast cancer cells and MCF-7 cells was detected.Results:The cytotoxicity of DC-Ag-TDLNC,DC-TDLNC and TDLNC to the auto-breast cancer cells was 67.64%、31.25% and 26.36% respectively,P0.05.Conclusion:Typical DCs can be induced from the mononuclear cells from axillary draining lymph nodes after stimulated with cytokines(rhGM-CSF,rhIL-4 and TNF-?).The DCs possess stronger antigen presentation and can obviously increase the killing activity of tumor antigen specific CTLs to auto-breast cancer cells.

Objective:The mouse mammary cancer cell line IL-23/MA-891 expressing IL-23 protein was set up and the effect of IL-23 on growth and other biological character of the cells was studied.Methods:The IL-23 gene was sequencely transfected into two packing cell lines (ecotropic ?2 and amphotropic PA317) by a retrovirus vector (LXSN),and the positive cellular clones were screened by G418.After infection to MA-891 cells with the culture supernatant of IL-23/PA317 cells and selected with G418,the IL-23/MA-891 cells to express IL-23 mRNA and protein was detected with RT-PCR,ELISA and immunocytochemical stairing,respectively.The expression of H-2Kb(MHCⅠ),I-Ab(MHC Ⅱ),CD80,CD86 and FAS was examined with flow cytometry.The ability to secrete IFN-? by mouse splenocytes stimulated with the culture supernatant of IL-23/MA-891 cells was detected with ELISA.The proliferation of MA-891,LXSN/MA-891 and IL-23/MA-891 cells was detected by MTT colorimetry and flow cytometry in vitro.Results:IL-23/MA-891 cells express IL-23 stably in mRNA and protein level.The expressing of H-2Kb(MHCⅠ),I-Ab( MHCⅡ),CD80,CD86 and FAS protein by flow cytometry were showing no difference in three kinds of cells.The proliferation rate of IL-23/MA-891 cells were showing little decresed,but statistically showing no difference with parental cells.The culture supernatant of IL-23/MA-891 cells induced secretion of IFN-? by mouse splenocytes were increased compared to parental cells.Conclusion:The cellular clone IL-23/MA-891 with high level expression of IL-23 was produced.

Objective To study the inductive effect of Cortex Periplocae extract(CPE) on apoptosis of human gastric cancer cells BGC-823 and its mechanism.Methods The cell morphology and super-microstructural changes of apoptosis were analysed by Giemsa staining and electric microscope,respectively.The BGC-823 apoptosis ratio,cell cycles,and changes of apoptosis in DNA level were studied by Flow Cytometry and agarose gel electrophoresis.The genes mRNA and protein expression of apoptosis-related genes bcl-2,bax,and survivin were studied by RT-PCR and immunology cell chemistry method.Results After treatment with CPE,BGC-823 cells showed some typical morphologic features and super-microstructural changes of apoptosis.DNA agarose gel electrophoresis showed characteristic "DNA ladder" pattern.Most BGC-823 cells were arrested at G_2/M phase.Some typical subdiploid peaks before G_0/G_1 phase were observed.The apoptotic rate of BGC-823 was 18.9% after 250 ?g/mL CPE treated for 48 h.The gene mRNA and protein expression of bcl-2 and survivin were inhibited by CPE,whereas that of bax was up-regulated.CPE could enhance the life span of S_(180) bearing mice in a dose-dependent manner.Conclusion(CPE can) inhibit the tumor growth by arresting the BGC-823 cell cycle at G_2/M phase and inducing BGC-823 apoptosis.Its mechanism is related to the inhibition on gene mRNA and protein expression of bcl-2 and survivin,and enhancement of those of bax.

Objective:To observe effects of Shenkui Decoction on proliferation of ovarian cancer cell.Methods:The Shenkui Decoction- containing rat serum was prepared by using TCM serum pharmacological method,and effects of the drug containing serum on proliferation of ovarian cancer fresh parenchymatous tumor cells were investigated with 3H incorporation method,and effects of the drug-containing serum on proliferation of ovarian cancer cell line Tyk-nu cells were investigated by flow cytometry,cellular activity assay,cellular growth curve assay and cell colony formation rate assay.Results:The drug containing serum could inhibit proliferation of both fresh parenckymatous tumor cells and ovarian cancer cell line Tyk-nu cells,and the inhibitory rate raised with the increase of drug-containing serum content.Conclusion:The Shenkui Decoction-containing rat serum can inhibit proliferation of ovarian cancer fresh parenchymatous tumor cells and ovarian cancer cell line Tyk-nu cells.