Objective To determine whether Rb gene is involved in the genesis of primary hepatocellular carcinoma(HCC). MethodsForty paraffin specimens of primary HCCs with corresponding adjacent liver tissues and normal liver tissues were investigated for Rb protein expression by tissue chip and SP immunohistochemical technique. ResultsLoss of Rb protein expression occurred in 17 of 40 tumor samples, whereas in 4 of 40 adjacent liver tissue samples, only 1 of 40 normal liver tissue specimens showed negative Rb staining.Rb protein deletion in HCC was higher than that in adjacent tissues and normal liver tissues (P＜0.05).Rb protein deletion rate doesn’t correlated remarkably with tumor size or phathology grade of HCC （Pgt;0.05）. ConclusionRb protein deletion may play an important role in the tumorigenesis of HCC.Tissue chip is an effective highthroughput technique platform for the study of tumor molecular pathology.
Objective To observe the expression and distribution of transforming growth factor-β1 (TGF-β1) in the healing process of bile duct and discuss its function and significance in the process of benign biliary stricture formation. Methods An injury to bile duct of dog was made and then repaired. The expression and distribution of TGF-β1 in the tissue at different time of the healing process were studied after operation with immunohistochemical SP staining. Results TGF-β1 staining was observed in the granulation tissue, fibroblasts and endothelial cells of blood vessels. High expression of TGF-β1 was observed in the healing process lasting for a long time. Conclusion The high expression of TGF-β1 is related closely with the fibroblast proliferating activity, extracellular matrix overdeposition and scar proliferation in the healing process of bile duct.
ObjectiveTo investigate the expression and relation of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in rats with diabetic retinopathy.MethodFifty-five Wistar rats were randomly divided into the control group (10 rats), and 1, 3, and 5-month-diabetes group (15 rats in each diabetes group), and the diabetic models were set up. The expressions of VEGF and bFGF were detected by situ hybridation and immunohistochemistry on retinal paraffin sections.ResultsThe results of situ hybridation showed that expression of bFGF was found in 3-month-deatbtes group with the percentage of 77.8%, and 88.9% in 5-month-deatbtes group; the positive expression of VEGF was not found in 3-month-deatbtes group but in 5-month-deatbtes group with the percentage of 66.7%. Immunohistochemistry indicated that the positive expression of bFGF started in 3-month-deatbtes group with the percentage of 55.6%, and 88.9% in 5-month-deatbtes group; the percentage of the expression of VEGF was 33.3% in 3-month-deatbtes group and 88.9% in 5-month-deatbtes group.ConclusionThe expression of VEGF occurs after the expression of bFGF in rats with DR.(Chin J Ocul Fundus Dis, 2005,21:37-40)
Objective To research the methods and techniques of SD rat bone marrow stromal cells （MSCs）culture in vitro and to provide a large number of MSCs for cell therapy. Methods Bone marrow from the femur and tibia of the early age SD rats was taken to culture to the passage 1-4 （P1-P4）, its growth was observed and P3 cells were evaluated by HE staining and immunohistochemisty. Results The growing speed of P1-P3 was faster than that of P0, cells fusion was 85%-90% after 3-4 d and the cells were arranged in groups liked whirlpool shape or parallel; （3.4-3.6）×104/cm2 cells were gained and the total cell number of P1-P3 was 4.08×106, 2.44×107 and 2.85×108 respectively, the rate of trypan blue rejecting stained was 95%-97%. P4’s growing speed was slower than before, 1.42×109 in 3.0×104/cm2 cells were gained, and the rate of rejecting stained was 95%. P4-cell output was amplified nearly 2 000-fold higher than P0-cell. P3 immunohistochemical analysis indicated CD105+ cells 61.9%， CD44+ 45.4%， CD29+ 16.8%， CD45+ 8.2%， CD31+ 13.7%， CD34+ 8.3% and CD11b+ 1.5%, respectively. Conclusion The culture of whole bone marrow is suitable for a large number of MSCs provision in vitro, and can meet the needs of the cell therapy research.
ObjectiveTo study the cause of different prognosis in typical medullary carcinoma and atypical medullary carcinoma. MethodsThe immunohistochemical staining method was used to evaluate the positivity of Ecadherin,βcatenin, in 30 cases of atypical medullary carcinoma and 18 cases of typical medullary carcinoma and 10 cases of normal breast. ResultsThe positive rate and staining intensity of Ecadherin and βcatenin oncoprotein were significantly higher in typical medullary carcinoma than in atypical medullary carcinoma(Plt;0.01).ConclusionExpression of Ecadherin and βcatenin is one of the causes of similar morphology and different prognosis in medullary breast carcinoma.
Objective To observe the expression of related proteins of retina after subretinal implantation with inactive chips.Methods A total of 27 healthy adult New Zealand white rabbits were randomly divided into three groups: operation group (12 rabbits) in which the rabbits were implanted with inactive chips into the interspace beneath retina;shamoperation group (12 rabbits) in which the rabbits were implanted with inactive chips into the interspace beneath retina which was taken out immediately;the control group (3 rabbits). Animals were sacrified for immunohistological study 7,15,30 and 60 days after surgery.The rabbits in control group group were sacrified for immunohistological study after bred for 30 days.The expressions of glial fibrillary acidic protein (GFAP) and brain derived neurotrophic facor (BDNF) were observed.Results In operation group, the outer nulear layer of retina thinned, and the cells in the inner nulear layer was disorganized 7,15,and 30 days after the surgery;glial cells proliferated 60 days after surgery; the positive expression of BDNF and GFAP was more than that in the shamoperation and control group.In shamoperation group, the positive expression of BDNF and GFAP was more than that in the control group.No obvious difference of expression of BDNF and GFAP between each time point groups was found.Conclusions The expression of neroprotective related proteins increased after subretinal implantation with inactive chips suggests that limited neuroprotective effects might be led by the implantation.
Objective To investigate the expression of Human leucocyte antigen(HLA)-DP, -DQ, -DR and CD40 in human retinal pigment epithelial (RPE) cells, to determine their molecule expression in immune response process, and their abilities to stimulate T lymphocyte activation. Methods Human RPE cells were cultured with or without (IFN respectively. Expression of HLA-DP, -DQ, -DR and CD40 was measured by immunohistochemical staining. Meanwhile, peripheral blood mononuclear cells (PBMC) were cocultured with RPE cells in vitro, and then the expression of activated lymphocytes CD69 was measured by fluorescence activated cell sorter(FACS). Results Expression of HLA-DP, -DQ, -DR and CD40 antigen were enhanced by gamma;-interferon inducement. Increasing amount of CD69 positive lymphocytes were found in the co-culture system of RPE cells and PBMC. Conclusion T-lymphocytes in the peripheral blood were activated by human RPE cells which is antigen presenting cells with immunological characteristics potential.
Objective To investigate the characteristic of the expression of facilitative glucose transporter 1 （Glut1） in bronchial aveolar carcinoma （BAC） and the relationship between expression of Glutl and fluorine-18 fluorodeoxyglucose （18F-FDG） uptake. Methods Twenty patients with BAC were imaged with 18F-FDG positron emission tomography （PET） before surgery. Maximum standard uptake value （SUVmax） and mean standard uptake value （SUVmean ） of tumor and standard uptake value of normal lung （SUVIung） were measured. The expression of Glutl was studied in paraffin sections by streptavidin peroxidase （SP） immunohistochemistry. Results All tumors of the patients could be detected by ^18F-FDG-PET. 18F-FDG uptake of tumor was higher than that of normal lung （SUV SUV and SUVlong were 3.09± 1.35, 2.37±1.03 and 0.39±0.09 respectively）. The expression of Glutl were positive in all tumors （73. 8%± 14. 8% of positive cell rate, 2. 8 ± 0. 9 grade of staining intensity）. Predominantly cytoplasm positive with weak staining intensity were observed in many tumors. Glutl negtive were observed in normal bronchial and lung parenchyma. Positive correlations were found among 18F-FDG uptake, Glut1 expression and tumor size （P〈0. 01）. Conclusion Glutl expression with peculiarity in BAC is correlate with 18F-FDG uptake.
Objective To investigate the possible interaction between the ras and p53 genes overexpression in thyroid carcinoma, and whether there is correlation between the ras and p53 overexpression and clinico-pathological criteria. Methods Thyroid lesions from eighty patients were examined for expression of ras and p53 genes by the LSAB immunohistochemistic method. Of these patients, 54 were diagnosed as malignant lesions and 26 benign nodular thyroid disorders. Results The positive immunostain rate for ras and p53 genes was 90.7%, 23.0% and 55.5%, 30.7% in carcinoma and benign lesions respectively with statistically significance between thyroid carcinomas and benign disorders (Ｐ＜0.05). Both ras and p53 overexpressions coexisted in 30 thyroid carcinomas and follow-up showed that 3 of them died and 5 of them had recurrence within 4 years.Conclusion Activation of ras gene and inactivation of p53 gene are cooperatively associated in thyroid tumorigenesis. The concurrent overexpression of ras and p53 could result in a poor prognosis.
Objective To investigate the expression of presenilin-2(PS2) and glutathione S transferase π(GSTπ) and their role in the prognosis and therapy of infiltrating ductal breast carcinoma. Methods The expression of PS2 and GSTπ in tumor tissues from 210 patients with infiltrating ductal breast carcinoma confirmed by pathologic examination and treated with modified radical mastectomy was examined by using LSAB immunohistochemical method. Results The expression rate of PS2 was 49.5%(104/210) and the expression rate of GSTπ was 48.1%(101/210). The grade of the postoperative 5-year survival rate and 10-year survival rate in four groups of 210 patients, from high to low, was the group 1 (PS2 positive expression/GSTπ negative expression), the group 2 (PS2 positive expression/GSTπ positive expression), the group 3 (PS2 negative expression/GSTπ negative expression) and the group 4 (PS2 negative expression/GSTπ positive expression). Conclusion The prognosis of the group 1 is the best, the group 2 better, the group 3 good and the group 4 the worst. The results suggest that reasonable use of endocrinotherapy and chemotherapy in infiltrating ductal breast carcinoma is necessary.