Objective:To verify whether hot PBS washing after the enzymatic reaction between tyramine and horseradish peroxidase can improve the sensitivity and specificity of catalyzed signal amplification (CSA). Methods: Using 7 different types of primary antibodies and 2 tumor microarray system, we carried out EnVision, LSAB, Standard CSA, and modified CSA staining and compared their sensitivities and specificities. Results: The modified CSA method was the most sensitive one among the 4 methods, followed by standard CSA, EnVision, and LsAB method. The sensitivity of modified CSA method was 2-3 times higher than that of standard CSA. The 4 methods had similar background staining and had exact localization of cells. Conclusion: The modified CSA is more stable and sensitive than traditional CSA staining method.

Objective:To apply chromogenic in situ hybridization(CISH)for detection of HER2 gene amplification in breast cancer tissues and to discuss some modifications of the CISH method.Methods:HER2 gene amplification was detected by CISH in 60 breast cancer specimens with an immunohistochemical score over 2+.The correlation between the results of IHC and CISH was analyzed.Our experience in CISH manipulation was summarized and optimization to CISH was discussed.Results:CISH identified gene amplification in 91%(40/44)specimens with an IHC score of 3+ and in 50%(8/16)specimens with an IHC score of 2+.The total concordance rate between IHC and CISH was 80%(48/60,P

The expressions of survivin,PcNA and Bcl xl were detected in 68 patients with pramary hepatic careinoma(HCC) and adjacent tissues and 12 cases of normal hepatic tissues by immunohistochemical (IHC) EnVision and Tunel method.The relationship between survivin gene expression and apoptosis index(AI),proliferation index(PI) and apoptosis inhibition factors were studied.The positive rate of survivin and Bcl xl expression were 58.8%(40/86) and 48.5%(33/68) in HCC,5.88%(4/68) and 36.8%(25/68) in adjacent tissues,0 (0/12) and 25%(3/12) in normal hepatic tissues.The expression of Survivin and Bcl xl are mainly in cell membrane and cytoplasm.Survivin expression is related to metastasis of HCC ( P

To investigate the relationship of expression of sialyl Lewis X (SLeX) antigen,CD44v6 and E cadherin(ED) with staging,infiltration,node metastasis and prognosis in breast cancer, immunohistochemical method was used to detect their expression in 94 cases of breast cancer.The positive expression of SLeX,CD44v6 and the negative expression of ED were positively correlated with the grading, clinical staging, lymph node metastasis recurrence and prognosis of breast cancer.The results demonstrated that the expression of SLeX, CD44v6 and ED as a clinical valuable marker,has a great significance in assessing node metastasis and prognosis.

Objective To characterize the localization and expression of cyclooxygenase (COX) in thoracic spinal cord before and after acute contusion. Methods 48 Sprague Dawley rats, 250 to 300 g in weight, were used for the study. The injuries of spinal cord were induced at the T8,9 level by dropping a weight of 10.24 g form a height of 50 mm (Allen’ s model). All the animals with spinal cord injury were sacrificed at time points ranging from 2 to 48 hours (2, 4, 8, 16, 24, and 48 hours) after management. Expressions of COX- 1 and COX- 2 in the thoracic spinal cords, normal and injured, were studied with immunohistochemistry. Results COX- 1 immunoreactivity was found to constitutively express in cytoplasmof glial cells and neuropils in white matter. Tiny COX- 1 immunoreactivity was found in glial cells, neuropils and neurons in the gray substance. It was found that COX- 2 immuno- labeling expressed constitutively in cytoplasm and closely surrounding the nucleus of neurons in both ventral and dorsal horns. Contusion to the spinal cord did not result in changes of COX- 1 protein localization and expression according to evaluation of immunohistochemistry. COX- 2 immunoreactivity improved only in neurons 4 hours following contusion. The positive COX- 2 protein reactivity reached the peak 24 hours after contusion. Conclusions In contrast to their expression in the majority of peripheral tissues, both COX isoforms express constitutively in the normal rat thoracic spinal cord. The major isoform of COX involved in the secondary spinal cord injury is COX- 2 after the spinal cord is mechanically injured.

Immunohistochemical localization of many sorts of tumor tissues from paraffin sections was studied using the ABC method with the monoclonal antibody secreted by the established two anti-osteosarcoma cell lines. It showed that the OS-McAb1 and OS-McAb2 had a negative reaction on the tested benign tumors and malignant tumors of epithelial tissue. Of the tested malignant tumors of mesenchyma, the antibodies had a positive reaction on some osteogenic sarcoma. They did not cross-react with various normal adult or fetal tissues, indicating that the OS-McAbs had a rather high specificity. They had a certain practical value in the immunopathologic diagnosis of malignant osteogenic tumors.

Objective: To study the expression of CD15 mRNA,CD44v6 mRNA and nm23H1 mRNA in colon carcinoma and its relationship with clinical pathological parameters and prognosis of colon carcinoma. Methods: In situ hybridization was used to detect the expression of CD15 mRNA, CD44v6 mRNA and nm23H1 mRNA in 90 cases of colon carcinoma, and 53 cases were followed up for more than 5 years. Results: In 90 cases of colon carcinoma, the expression of CD15 mRNA,CD44v6 mRNA and nm23H1 mRNA were 84.4%,68.9%,66.7%, respectively. The high level expression of CD15 mRNA, CD44v6 mRNA and the low level expression of nm23H1 mRNA were positively corelated with the Dukes staging,serosa infiltration,lymph node and liver metastasis of colon carcinoma.The expression of CD15 mRNA in colon carcinoma was positively associated with that of CD44v6 mRNA and negatively with that of nm23H1 mRNA. Conclusion: CD15 mRNA,CD44v6 mRNA and nm23H1 mRNA play an up-regulation and a down-regulation role, respectively, and have a synergistic action in metastasis of colon carcinoma. CD15 mRNA can be used as a new biological index to predict the invasive potential of colon carcinoma and objectively evaluate the prognosis of patients.

Objective: To investigate the pathological changes of human hepatocellular carcinoma (HCC) after continuous passaging in nude mice. Methods: The mice model of HCC SMMU-LTMN were continuously observed for 20 years (1987-2007) . The subcutaneously transplanted carcinoma had been passaged for 228 generations. The pathological data of abdominally transplanted HCC, orthotopically transplanted HCC in nude mice, and orthotopically transplanted HCC in NOD-SCID mice were recorded. The pathological studies were conducted by light microscope, electron microscope, image analysis, chromosomal analysis, and measurement of alpha-fetoprotein (AFP) in peripheral blood. Results: (1) The local invasion and metastasis of of tumors were present in all the above 4 models for a long time. The local invasion rate and the pulmonary metastasis rate of subcutaneously transplanted tumors were 59.70% (40/67) and 37.10% (23/62), respectively. The pulmonary metastasis rate of abdominally transplanted tumors was 59.02%(36/61). The intra-hepatic and pulmonary metastasis rate of the othotopically transplanted tumors were 18.18%(4/22) and 31.82% (7/22), respectively. The pulmonary metastasis rate of HCC in NOD-SCID mice was 53.85%. (2) The tissue structure and the differentiation of the 10th generation tumor cells was similar to those of primary HCC, with grade 2 differentiation and coarse trabecular pattern as the main characteristics. From the 11th generation to the 228th generation, the main characteristics of tumor cells were grade 3 differentiation and lump pattern. Electron microscope also showed worse differentiation. (3)The AFP level was 92 500 ?g/L in cells before the 32th generation; it decreased to 6 729?g/L from the 33th-130th generation cells; and the level of the 220th generation was 1 000-5 000 ?g/L.(4)The DNA contents had a wide distribution (from 2c to 6c) in abdominally transplanted tumors and the pulmonary metastatic tumors; the mean DNA index in the former tumors (2.60?0.20) was wider than the that in the latter (2.10?0.26) . (5)From the 55th generation to 206th generation, it was found that tumor cells had integrated into the chromosome of the nude mice. Conclusion: The subcutaneously transplanted HCC in nude mice can be stably expressed for 20 years, with no change in the local invasion and metastasis ability of HCC. The differentiation of the tumor cells worsenes and the AFP level is decreased in the blood; some chromosome of tumor cells integrate into the chromosome of nude mice, which may be related to the internal environment of nude mice and the multi-potential differentiation of the tumor cells.

To investigate the role of intercellular adhesion molecule 1(ICAM 1) in the pathogenesis of bacterial keratitis,22 rats were randomly divided into bacterial keratitis group and control group. A specific staining of corneal epithelial cells, stromal keratocytes and endothelial cells for ICAM 1 was carried out by using monoclonal antibody to ICAM 1 and immunohistochemical staining techniques. The results showed that ICAM 1 expressed a low level in the corneal epithelial cells, stromal keratocytes and endothelial cells in the control group(-~+), while its expression was strong in the keratitis group (~). The difference between two groups was statistically significant ( P

Objective To discuss the growth suppressing, apoptosing effect of new type tumor-supressor gene-p33ING1 in stomach cancer cell strain, and to explore new strategies and methods in tumor therapy. Methods The PCDNA3/p33ING1 nuclear expressing microsome was constructed, p33ING1 and wt-p53 were implanted to human stomach cancer cell both and to evaluate the effect of p33ING1 and p53 toward stomach cancer cell and synergism between them. Results The PCDNA3/p33ING1 nuclear expressing microsome was successfully constructed. The human stomach cancer cell strain SSCG-7901 under implantation of p33ING1 and wt-p53 showed a significant decrease in cell growth, the coupling time was delayed, DNA synthetic phase was shortened and G0/G1 phase prolonged. The cell collapse increased. Conclusions Despite of the tumor-inhibiting effect and biochemical activation of p33ING1, it also plays a role with p53 gene in controling growth of stomach cancer cell, inducing cell collapse and hampering cell proliferation cycle. P33ING1 and p53 are synergistic to each other.