4.Development and application of stem cells.
Chinese Medical Journal 2005;118(10):795-798
5.Expression profile of inflammation related genes in laryngeal cancer
Lin LI ; Dongdong ZHU ; Zhen DONG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(02):-
OBJECTIVE To investigate the expression profile of inflammation related genes in laryngeal squamous cell carcinoma by functional gene chip technique and to probe into the role of correlative genes in pathogenesis of the laryngeal squamous cell cancer and in tumor immunity. METHODS The total RNAs were respectively extracted from two pair samples of laryngeal tumor and the normal tissue around the tumor, and then were reversely transcribed to cDNAs, then synthesized to cRNAs. The cRNAs were labeled with the hybridization probes. The probes were then hybridized with four pieces of inflammation related genes chip. It was chemiluminescently detected and the acquired image was analyzed with special software. RESULTS Forty genes were differently expressed in inflammation related gene profile of laryngeal tumor, among which 22 genes were upregulated and 18 genes were down regulated. Thirteen genes were shown differential expression in both chips with 10 upregulated genes and 3 downregulated genes. CONCLUSION The differently expressed genes in inflammation related gene chips will provide clues and theoretical foundation for the investigation of the relationship between tumor and inflammation, and also the immune pathogenesis of laryngeal tumor. Furthermore CCL-7 may have an important role in the occurrence of laryngeal cancer, and the role of immunity of the pathogenesis of laryngeal tumor needs further studied.
6.Effects of murine cytomegalovirus infected mouse embryonic fibroblasts on the proliferation and activation of co-cultured regulatory T cells
Yanan LI ; Feng FANG ; Hong ZHEN ; Yufeng ZHOU ; Yongsui DONG
Chinese Journal of Infectious Diseases 2008;26(11):641-646
Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P<0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P<0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.
7.The relationship between ImmuKnow value of cellular immune function and peripheral white blood cells count and T-lymphocyte after liver transplantation
Ruidong LI ; Jiayong DONG ; Zhen SUN ; Zhiren FU ; Zhengxin WANG
Chinese Journal of General Surgery 2011;26(6):506-508
Objective To explore the relationship between peripheral differential blood count and ATP value in Cell CD4 + T tested by ImmuKnow method in liver transplants. Methods In this study 49recipients after classic orthotopic liver transplantation (OLT) were enrolled. In a period from two weeks to two months after transplantation when all were free of glucocorticoid. Blood were sent for WBC differential samples count and ATP value in Cell-CD4 + T tested by ImmuKnow method via SPSS17. 0 software. Five more samples were selected randomly for duplicated testing of the indices in Week2, 3, 4,6 and 8 after the transplanting operation to further verify the relativity. Results White blood cell count has the highest relativity with ImmuKnow ATP value at 0. 821. The 5 recipients were repeatedly tested for ImmuKnow ATP values that were found positively correlated to cell count with a coefficient of over 0. 5. Conclusions The peripheral leukocyte count in early stage after liver transplantation is in positive correlation with ATP value in Cell CD4 + T, and the changes of numeration of leukocyte reflect changes of ATP value.