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.

OBJECTIVETo investigate the clinical pathology of cavernous venous malformations of the body surface.

METHODSTissue samples of cavernous venous malformations from 42 cases were stained with hematoxylin and eosin to observe the pathologic structure. The clinical manifestations and case history were summarized accordingly.

RESULTSThere was no distribution difference of the malformation in sex and body sides, but with obvious difference in anatomic sites. The malformation occurred most frequently at the head and neck, more frequently at extremities and least frequently at the trunk. According to pathologic structure, cavernous venous malformations of the body surface can be divided into three types: the cellular, the canaliform and the mixed.

CONCLUSIONThe cause of distribution difference in anatomic sites remains unclear. Internal hemorrhage and infection may account for the increased growth and ache of the lesion. The different pathologic structure of the malformation may cause different clinical manifestations.

Adolescent ; Adult ; Aged ; Arteriovenous Malformations ; complications ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infection ; etiology ; Male ; Middle Aged ; Pain ; etiology ; Sex Factors ; Skin ; blood supply ; pathology ; Veins ; abnormalities

OBJECTIVETo investigate the expression of five kinds of cyclins in hepatocellular carcinoma (HCC) and their association with degree of tumor differentiation, metastasis and infection of hepatitis B virus (HBV).

METHODSThe HCC tissue microarrays were composed of those from 273 cases of HCC tissues, 144 surrounding-tumor liver tissues and 10 normal liver tissues obtained from autopsy. The diameter of each specimens on tissue microarrays was 2.0 mm. Immunohistochemistry was used to detect the expression of cyclin A, cyclin B, cyclin D1, cyclin D3 and cyclin E on HCC tissue microarrays. The association of the expression of these cyclins with the infection rate of HBV was also analyzed.

RESULTSThree paraffin-embedded HCC tissue microarrays were successfully constructed, including 136, 143 and 148 tissue spots, respectively. The positive rates of cyclins in 273 cases of HCC were cyclin A 52.7%, cyclin B 45.4%, cyclin D1 35.9%, cyclin D3 44.3% and cyclin E 23.1%; while the figures in 144 surrounding-tumor tissues were 8.3%, 5.6%, 4.9%, 6.3% and 1.4%, respectively. In 10 normal liver tissues these cyclins exhibited negative staining, with the exception that cyclin D1 was positive in one case of normal liver tissue. The positive rate of cyclins in HCC were significant higher than those in surrounding-tumor liver tissues (P < 0.01), in HCC tissues with histological grade II and III, the cyclins expression were stronger than that in grade I (P < 0.05). The positive rates of cyclins, except cyclin A in HCC with portal vein invasion were higher than those without portal vein invasion (P < 0.01). Infection of HBV did not have significant relationship with the expression of cyclins (P > 0.05).

CONCLUSIONCyclins in different cell cycles overexpressed at varied levels in hepatocellular carcinoma, and the increased expression of cyclins may shorten the tumor cell cycle phase, accelerate cell proliferation, and have a close relationship with HCC aggressiveness.

Carcinoma, Hepatocellular ; chemistry ; Cyclin A ; analysis ; Cyclin B ; analysis ; Cyclin D1 ; analysis ; Cyclin D3 ; Cyclin E ; analysis ; Cyclins ; analysis ; Hepatitis B ; metabolism ; Humans ; Immunohistochemistry ; Liver Neoplasms ; chemistry

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