To search for the relationship between immune state and tumor metastases, CD3,CD4,CD8 and CD44 contents in 13 speciments of fine needle aspiration (FNA) from thyroid cancer patients were detected by the flowcytometry (FCM) and comparison between thyroid cancer and benign tumor was made. The result showed :in thyroid cancer group, CD＋3，CD＋4 cells and the ratio of CD＋4/CD＋8 reduced significantly (Ｐ＜0.01),and CD＋8 cell increased significantly (Ｐ＜0.01), in metastases group,this change was much significantly. CD44 expressed significantly higher in cancer group than that of the benign thyroid neoplasms, and the change was related to the tumor metastases. The results indicate that CD44 can be as a marker of tumor and indicator of metastases. The disturbance of immune system results in active expression of CD44 by tumor cells, so, lead to metastases. It is helpful to the diagnosis of thyroid cancer, assessment of metastases and management in surgery.
Objective To observe the proapoptotic effect ofthe homogenate of different parts of pig’s full thickness dermal wounds on cultured fibroblasts. Methods The tissues were dissected from the wound center and subneoepithelium separately 15 days after homogenization and sterilization, the specimens stored at -70℃. The forth passage of the fibroblasts were cultured for 16 hours in different culture solutions and were grouped into 7 groups: DMEM containing 5% fetal bovine serum as Group Ⅰ, DMEM containing 5% homogenate of tissue from wound center as GroupⅡ, DMEM containing 5% homogenate of tissue from subneoepithelium as Group Ⅲ, the culture solution of Group Ⅱmixed with 10 μg/ml GM6001 in Group Ⅳ, with the culturing medium of Group Ⅲplus 10 μg/ml GM6001 as Group Ⅴ, the culture solution of Group Ⅱ mixed with 10 ng/ml aFGF as Group Ⅵ, and the culture solution of Group Ⅲ mixed with 10 ng/ml aFGF as Group Ⅶ. In all groups except Group Ⅰ, the fibroblasts of the 6 pigs were treated with the homogenate derived from the same animal respectively. After being incubated in Annexin Ⅴ-FITC and PI, cells were analyzed by Flow Cytometry and the rate of apoptotic cells was acquired. The data were analyzed by SPSS 11.0 using Leastsignificant Difference test(LSD). Results The apoptotic rate of the 7 groups were as follows:4.39%±0.41% in Group Ⅰ，10.98%±1.42% in Group Ⅱ，13.47%±1.44% in Group Ⅲ，7.2%±0.46% in Group Ⅳ，12.1%±0.85% in Group Ⅴ，3.9%±0.63% in Group Ⅵ，9.8%±0.50% in Group Ⅶ； there were significant differences between every two groups except Group Ⅰand Group Ⅵ. Conclusion Homogenate of the tissue derived from the subneoepithelium has greater proapoptotic effect than that from the wound center; the proapoptotic effect of homogenate of the tissue both under neoepithelium and in wound center can be significantly alleviated by acid fibroblast growth factor, partly because of MMPs.
For the purpose of understanding the distribution of insulin-like growth factor-1 (IGF-1) receptor on the tendon cell, the continuous cultured tendon cell line was studied by following experiments. With the methods of immunohistochemical study and flow cytometric study, the density of IGF-1 receptor of the primary, 6th and 13th generation of tendon cell was analyzed. The results showed that there was no difference of the receptor density among those generations. However, in the cell cycle, the numbers of IGF-1 receptor in G2M phase tendon cells were more than that in G1 phase cells (P lt; 0.01). These works provided sufficient evident which suggested there were stable density of IGF-1 receptor on the tendon cell though out the life span of tendon cell. This may build some foundation in growth control of tendon cell by growth factor in the research of tendon tissue engineering.
目的 比较使用流式细胞仪355 nm和407 nm激光器激发Hochest33342检测细胞凋亡。 方法 通过ATO药物诱导急性早幼粒白血病细胞（NB4）及血清饥饿法诱导人肺癌细胞（NCl-H292）细胞凋亡，取24、48 h时间点收集细胞，进行Hoechst33342-碘化丙啶（PI）双染，分别在配置有两种激光器的流式细胞仪上检测细胞凋亡。 结果 细胞经处理后24 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（28.20 ± 4.80）%；NCl-H292细胞凋亡率Hoechst33342+/PI-：（22.47 ± 2.78）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（25.10 ± 6.19）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：20.47 ± 1.46%。处理后48 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（33.60 ± 3.75）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（26.77 ± 1.16）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（29.47 ± 2.33）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（31.47 ± 3.05）%。两种细胞处理后比处理前凋亡率明显升高，但355 nm激光器与407 nm激光器检测的凋亡结果差异不明显（P＞0.05）。 结论 407 nm激光器激发Hoechst33342可检测细胞凋亡。
【Abstract】ObjectiveTo find a stable and efficient method for ex vivo concentration of CD4+CD25+ regulatory T cells in rats. MethodsCD4+CD25+ regulatory T cells were separated from the rat splenic cells in two steps by magnetic cell sorting (MACS) system. CD4+ T cells were first negatively sorted by cocktail antibodies and antiIgG magnetic microbeads. And then CD4+CD 25+T cells were positively sorted by antiCD25 PE and antiPE magnetic microbeads. The purity and the cell survival rate of the sorted cells were measured by flow cytometry and trypan blue dyeing respectively, and the immunosuppressive action of CD4+CD25+ T cells on the proliferation of CD4+CD25- T cells was also assessed by in vitro cell proliferation assay. ResultsThe purity of negatively sorted CD4+ T cells were (83.6±2.5)% (79%~87%), and the purity of positively sorted CD4+CD25+ T cells was (90.2±1.8)% (86%~93%) with the survival rate of (92.8±3.4)% (92%~95%). These concentrated cells significantly suppressed the proliferation of CD4+CD25- T cells in mixed lymphocyte culture （CD25+/CD25- versus CD25-, P＜0.01）. ConclusionWe created a twostep procedure of magnetic cell sorting system for CD4+CD25+ regulatory T cells sorting, which insures the cells to be satisfactorily purified and well functioned.
In order to solve the problem of the micro flow cell clogging, and to improve the reliability of the flow cytometry system, a new method was proposed for hydrodynamic self-cleaning system. By analyzing the flow cell focus principle, we considered that to obtain stable single cell flow, the stable pressure in the flow chamber must be ensured. Therefore, we established a diagnosis method of clogging by the pressure detecting, and designed a self-cleaning system. Then we built up corresponding experimental systems. Experiments and testing showed that the self-cleaning system could improve the flow and resolve the clogging problem.