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<0.01). The thickness of sections should be controlled within 4-5 μm; the denaturation should be complete; and the post-hybridization washing temperature and time were also very important and the temperature should be controlled at 70-75 °C. The dyeing time of hematoxylin should also be restrictedly controlled. Positive control should be set up in the experiment for high quality of the experiment. Conclusion: CISH has high concordance rate with IHC in examining HER2 amplification and it may be a new method for detection of HER2 gene. The thickness of the sections, the post hybridization washing temperature and time, and the time of hematoxylin dyeing should be strictly controlled.

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 En Vision, 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, En Vision, 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.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Female ; Homeodomain Proteins ; biosynthesis ; genetics ; Humans ; Middle Aged ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Estrogen ; metabolism ; Transcription Factors ; biosynthesis ; genetics

Objective To investigate the nerve recanalization and the motor function of hind legs after transplantation of peripheral nerve grafts treated with microsurgical technique at chronic spinal cord injury (SCI) in rats.Methods The SD rats were established into SCI model with improved Allen method.The rats were divided into two parts 12 weeks after the injury.In experimental group:by microsurgieal technique. the sural nerves were removed epineufium and transplanted into SCI lesion,control group rats were treated without any operation.Retrograde HRP tracing through sciatic nerve were practiced at 1 month,2 month,3 month after transplantation of peripheral nerve grafts.The morphological changes were observed at section of spinal cord and the motor functions of both hind legs of rat were detected.Results The morphology of the injured spinal cord sections turned better.Retrograde HRP tracing through sciatic nerve showed some HRP positive markers at the site of near rostral end of the nearly injured part at one month after transplantation and increased with the time going by.Motor function of hind legs of rats recovered significantly in transplantation groups.Conclusion Peripheral nerve grafts treated with mierosurgical technique have repairing effect on chronic spinal cord injury in rats.

Actins ; analysis ; Antigens, CD34 ; analysis ; Carcinoma, Papillary ; classification ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Keratin-19 ; analysis ; Keratin-20 ; analysis ; Muscle, Smooth ; chemistry ; Pancreatic Neoplasms ; classification ; metabolism ; pathology ; Proto-Oncogene Proteins c-kit ; analysis ; Receptors, Estrogen ; analysis ; Receptors, Progesterone ; analysis

Objective To observe the effects of culture medium of a mn iotic cells on NO and NOS in retinal tissues of rabbits in vitro in order to pro vide a protective method for antioxidation in retina transplantation. M ethods Thirty adult healthy rabbits (30 right eyes) were divided into 3 groups. Group Ⅰ: fresh retinal tissue; group Ⅱ: routine culture medium; group Ⅲ: culture medium of amniotic cells. The retinal tissues in group Ⅱ and Ⅲ we re cultured in the corresponding culture medium for 1 week. The content of NO an d NOS in retinal tissues in the 3 groups were determined. Results Compared with group I, the content of NO and NOS of group Ⅱ increased obvio usly (t=3.821, 3.854; P0.05) . Conclusion Culture medium of amniotic cells may remove free r adicals and enhance the ability of antioxidation.

0.05).Conclusion MK mRNA overexpresses in the breast cancer tissues.It might be considered to be a reference indicator for deter- mining the angiogenesis and invasion of breast carcinoma.

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; virology ; Genes, p53 ; Hepatitis B ; genetics ; metabolism ; virology ; Hepatitis B virus ; Humans ; Inhibitor of Growth Protein 1 ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; virology ; Loss of Heterozygosity ; Nuclear Proteins ; metabolism ; Point Mutation ; Signal Transduction ; Trans-Activators ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo investigate the expression levels of nm23, P53, and S100A4 in gastric carcinoma and their relationships with clinicopathologic parameters and metastasis potential.

METHODSPathological specimens from gastric carcinoma,matched para-tumor tissues, metastatic lymph node and distant metastatic tissues were examined for the expression levels of nm23, P53, and S100A4 proteins by tissue microarray technique and immunohistochemistry.

RESULTSThe expression levels of P53 and S100A4 were upregulated (P< 0.01), while the expression of nm23 downregulated (P< 0.05) in gastric carcinoma compared with non-tumor tissues. S100A4 expression was significantly higher in distant metastatic tissues, while nm23 lower in metastatic lymph nodes than those in cancer tissues. Upregulating expression levels of nm23, P53, and S100A4 were significantly correlated with some malignant behaviour of gastric cancer. The expression rates of nm23+/P53+, P53+/S100A4+, and nm23+/S100A4+ immunohistochemical phenotypes were 48/74 (64.9%), 50/74 (67.6%), and 39/74 (52.7%). P53+/S100A4+, nm23+/S100A4+, and nm23+/P53+/S100A4+ phenotypes were associated with high metastasis potential of gastric cancer.

CONCLUSIONSAlteration of nm23, P53, and S100A4 expression may contribute to the development of gastric carcinoma. Nm23 and S100A4 proteins play a critical role in tumor metastasis. Co-detection of the expression of P53, nm23, and/or S100A4 can be used to evaluate high metastasis potential of gastric cancer.

Female ; Gene Expression ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; metabolism ; Neoplasm Invasiveness ; Neoplasm Staging ; S100 Calcium-Binding Protein A4 ; S100 Proteins ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVETo explore the pathological mechanism in the repair of chronic spinal cord injury with free grafting of autoperipheral nerve tissues in rats.

METHODSThe SD rats were used to establish SCI model with modified Allen method. The rats were divided into two groups at 12 weeks after the injury, each group had 20 rats. In the experimental group, the sural nerves were removed epineurium and transplanted into SCI lesion by using microsurgical technique; and in the control group, the rats were treated without any operation. The survival and differentiation of the grafts, and the ability of repairing host spinal cord were observed under the light microscope at the postoperative 4th and 12th week. Regeneration rates of nerve tracts in spinal cord were evaluated by using HRP tracing technique at the postoperative 4th and 12th week. The morphological changes were observed at section of spinal cord and the motor functions of both hind legs of rats were detected.

RESULTSIn the control group, spinal cord exhibited degeneration with cicatrices and cavitates. In the experimental group, peripheral nerve was almost survived, fused with the spinal tissue and axons could regrow into or span the place of injured spinal cord. Higher number of labeled nerve tracts in spinal cord were observed in experimental group, there was significant difference when compared with the control group. Motor function of hind legs of rats recovered significantly in the treatment group.

CONCLUSIONAutoperipheral nerve graft tissues transplantation could survive and integrate with the host and have repairing effects on chronic spinal cord injury in rats.

Animals ; Female ; Male ; Peripheral Nerves ; transplantation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; physiopathology ; surgery ; Transplantation, Autologous