Objective To observe whether Cyclo-RGDfK (Arg-Gly-Asp-D-Phe-Lys) could enhance the adhesion of myofibroblast to decellularized scaffolds and upregulate the expression of Integrin αVβ3 gene. Methods Myofibroblast from the rat thoracic aorta was acquired by primary cell culture. The expression of Vimentin and α-smooth muscle actin(α-SMA) has been detected by immunoflurescent labeling. Decellularized valves have been randomly divided into three groups (each n=7). Group A (blank control): valves do not receive any pretreatment; Group B: valves reacted with linking agent NEthylN(3dimethylaminopropyl)carbodiimide hydrochloride (EDC) for 36 hours before being seeded; Experimental group: Cyclo-RGD peptide has been covalently immobilized onto the surface of scaffolds by linking agent EDC. The fifth generation of myofibroblast has been planted on the scaffolds of each group. The adhesion of myofibroblast to the scaffolds was evaluated by HE staining and electron scanning microscope. The expression of Integrin αVβ3 was quantified by halfquantitative reverse transcriptionpolymerase china reaction (RT-PCR). Results We can see that myofibroblast has exhibited b positive staining for Vimentin and α-SMA. Besides, it has been shown that the expression of Integrin αVβ3 was much higher in the experimental group than that of the group A and group B(Plt;0.05). There was no statistically difference in group A and group B (P=0.900). Conclusion RGD pretreatment does enhance the adhesive efficiency of seeding cells to the scaffolds and this effect may be related to the upregulation of Integrin αVβ3.
OBJECTIVE To investigate the adhesive interactions of cells with materials and the effects of material properties on cell adhesion in tissue engineering. METHODS By looking up the recent literatures dealt with adhesive interactions of cells with materials and reviewing previous work on the adhesion of tissue-derived cells to materials. RESULTS The adhesion characteristics of cells to materials not only depend on the nature of materials, including bulk and surface properties, surface modification, surface morphology, net charge, porosity and degradation rate, but also on the expression of cell surface molecules and their interaction with the material. CONCLUSION The quantitative measure and biophysical mechanisms of cell adhesion to materials might be very important in tissue engineering.
OBJECTIVE: To investigate the influence of basic fibroblast growth factor (bFGF) on adhesion characteristics of osteoblasts, aimed at the important problem in bone tissue engineering of how to promote the adherence of osteoblasts to extracellular matrix materials. METHODS: 5 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml bFGF were used to induce bone marrow stromal-derived osteoblasts of rabbit for 24 hours before incubation, and the common culture medium as the control. The attached cells were calculated with stereology method at 0.5 hour, 1st hour, 2nd hour, 4th hour, 8th hour after seeding. RESULTS: The number of attached cells was significant higher in the experimental group when induced by 10 ng/ml bFGF than that in the control group (P lt; 0.01); the number did not increase with the increase of bFGF concentration and there was no significant difference between the experimental group induced by 100 ng/ml bFGF and control group, and the number was even obviously lower in the experimental group when induced by 200 ng/ml than the control group (P lt; 0.01). CONCLUSION: bFGF can influence the adhesion characteristics of osteoblasts, 10 ng/ml bFGF can promote the adherence of osteoblasts to matrix materials, but 200 ng/ml bFGF may inhibit cell adhesion.
To study the relationship between the expression and contents of cell adhesion molecule CD15 and differentiation and lymph nodes metastasis of breast carcinomas, CD15 expression and its contents in 94 cases of breast carcinomas and or cases of normal breast tissue were evaluated by microwave-SP immunohistochemical chenique combined with image analysis. CD15 immunoreactivity in normal breast tissue was mainly localised at the border of gland, but in breast cancer tissues it was mainly localised in the membrane and cytoplasm. Positive rate of CD15 and its average optic density in breast carcinomas were significantly higher than those in normal breast tissue (Ｐ＜0.05 and Ｐ＜0.001, respectively). The worse tumors differentiated and the earlier lymph nodes metastasized, the higher CD15 expressed and its optic density was measured (Ｐ＜0.05 and Ｐ＜0.001, respectively). These results suggest that CD15 expression and its contents might be a useful indicator to evaluate the malignancy and biological features, and could be considered as a good prognostic predictor for breast carcinomas.
Objective To review recent studies in molecular biology of gastric cancer. Methods Relevant references were reviewed. Results The development and progression of gastric cancer were correlated with oncogenes, growth factors, cyclins, tumor suppressor genes, cell adhesion molecules and unstability of genes.Conclusion Gastric cancer is related to much mutation of genes.
Objective To observe the effect of hypoxia inducible factor-1alpha;（HIF-1alpha;）to the expression of cell surface adhesion molecules CD18 and the adhesion ability of leukocytes and vascular endothelial cells under early stage of diabetic retinopathy condition.Methods The human promyelocytic leukemia cell line HL60 and the rhesus choroid-retina vascular endothelial cell line RF/6A were cultured in RPMI 1640 medium-10% human serum, which was collected from the subjects of early stage of diabetic retinopathy and age-matched healthy control. The cells were cultured in 4 groups as control group (group A), diabetic group (group B), HIF-1 anti-sense oligonucleotides (ASODN) group (group C) and HIF-1 sense oligonucleotides (SODN) group (group D). The percentages of CD18 positive cell in the HL60 cell were measured by flow cytometry and mRNA in the HL60 cell by realtime reverse transcriptionpolymerase chain reaction（RT-PCR）. Results The percentage of CD18 positive cell in the group A, B, C and D was 17.06plusmn;6.01, 42.23plusmn;2.60, 25.33plusmn;3.05 and 32.40plusmn;10.57, respectively, the differences among them were significant (F=36.47,P＜0.001). Compared to the group A,the expression of CD18 mRNA in the group B,C and D was increased about 21.05plusmn;2.07、2.23plusmn;0.96 and 25.07plusmn;2.27 times,respectively, the differences among them were significant (F=180.34, Plt;0.001). The adherent rates of HL60 to RF/6A in group A, B, C and D was 0.06plusmn;0.00,0.09plusmn;0.10,0.05plusmn;0.00 and 0.07plusmn;0.01, respectively,the differences among them were significant（F=13.06，P=0.002）.Conclusion In vitro, HIF-1 could regulate the expression of CD18 by HL60, and the adhesion of HL60 to RF/6A when the cells were exposed to diabetic serum. The effects of human serum weaken with the inhibition of HIF-1 expression.HIF-1 play regulatory role in the expression of CD18 and adhesion of leukocytes and vascular endothelial cells under early stage of diabetic retinopathy condition.
Objective To investigate the influence of CO2-insufflation pressure on invasion potential of the colon cancer cells. Methods With an in vitro artificial pneumoperitoneum model, SW1116 human colon cancer cells were exposed to CO2-insufflation of 5 different pressure groups: 6, 9, 12, 15 mm Hg and control group, respectively for 1 h. The invasion capacities of SW1116 cells exposed to CO2-insufflation of 5 different pressure groups were detected by cell adhesion/invasion assay in vitro. Results Immediately following exposure to 15 mm Hg CO2 insufflation, the invasion of SW1116 cells decreased significantly compared to the cells before exposure. At the 0 h time point, the cells exposed to 15 mm Hg were significantly less invasive than those exposed to the other insufflation pressure (P＜0.05), and the cells exposed to 6 mm Hg were more invasive than cells exposed to the other insufflation pressure (P＜0.05). And 72 h after exposed to CO2-insufflation, the differences between the pressure groups were not significant. Conclusion CO2-insufflation induced a temporary change in the invasion capacity of cancer cells in vitro, higher pressure of CO2-insufflation inhibits the invasion potential.
Objective To investigate the effect of interleukin-10 （IL-10） gene transfer on expression of CD44, selectin-E, lymphocyte function associated antigen-1 （LFA-1）, vascular cell adhesion molecule-1 （VCAM-1） in mice heart transplantation rejection. Methods Model of mice cervical heterotopic heart transplantation was set up, 96 mice were divided into three groups with random number table, control group： heart transplantation between C57 mice; transplant group： heart from BALB/C mice transplant to C57 mice; IL-10 group： IL-10 was transfected on BALB/C mice isolated heart for 1 hour, then transplanted to C57 mice. The messenger ribonucleic acid （mRNA） level expression of CD44 ,selectin-E ,LFA-1 ,VCAM-1 and IL-10 were measured by reverse transcription-polymerase chain reaction （RT-PCR） at the 5th day after transplantation. Results The mRNA level expression of CD44, selectin-E ,LFA-1 ,VCAM-1 in transplant group were significantly increased than those in control group （P〈0.01）. The mRNA level expression of CD44, selectin-E, LFA-1 ,VCAM-1 in IL-10 group were significantly decreased than those in transplant group （P〈0.01）. Conclusion IL-10 gene transfer is able to decrease the expression of CD44, selectin-E,LFA-1 ,VCAM-1 and suppress the heart transplantation rejection in mice.
ObjectiveTo summarize the progress of p120-catenin (p120ctn)——a new member of catenin family in tumor research. MethodsDemestic and international published literatures related to p120ctn in recent years were collected and reviewed. Results① p120ctn was involved in formation of cadherincatenin complex, participated in cell growth, proliferation, and adheren junctions. ② p120ctn regulated Rho GTP activity and promoted cell motility. ③ p120ctn was involved in the regulation of gene transcription through binding with the nuclear transcription factor Kaiso. ④ p120ctn was involved in angiogenesis process induced by vascular endothelial growth factor. ⑤ p120ctn was involved in inflammation, cell malignant transformation, and tumor invasion and metastasis. ConclusionsAs a new member of catenin family, p120ctn participates in a variety of biological processes relying on its cellular localization. It will be facilitated to judge the genesis and progression of tumor from the abnormal alteration of p120ctn according to understanding the biological function and mechanism of p120ctn in the molecular level, a new pathway in the prevention and treatment of cancer is provided.
Objective To investigate the level of cell adhesion molecule sialyl-LeX expression in colorectal carcinoma and its relation with carcinogenesis, differentiation, metastasis and prognosis. Methods Sialyl -LeX expression and its optical density in colorectal carcinoma (n=90) and remote normal mucosa (n=30) were quantitatively studied with microwave-LSAB immunohistochemical method combined with image analysis technique. Fifty-three patients were followed up. Results The weaker staining in remote normal colorectal mucosa was observed in very limited parts of some deep crypts. Positive rate of sialyl-LeX expression was only 16.7%(5/30). The positive expression of sialyl-LeX was observed in 83 of 90 patients with colorectal carcinoma(92.2%). The apical cytoplasma of cancer tubules, the luminal contents, and the cytoplasma of the cancer cells were bly stained. The mean integral optical density of sialyl-LeX positive cell in poorly differentiated adenocarcinoma was significantly higher than that in highly differentiated and mucinous ones (Plt;0.01). It was markedly higher in patients with positive lymphatic nodes than that in negative ones (Plt;0.01). With followed-up for longer than 5 years, it was much lower in the alive cases than that in the dead (Plt;0.01). Conclusion These findings indicate that changes of sialyl-LeX expression and its optical density is related to carcinogenesis, differentiation, invasion and metastasis of colorectal carcinoma. It may be a good predicter for the prognosis of patients with colorectal carcinoma.