Breast neoplasm shows high rate of incidence and mortality among females.Although thereare a variety of treatments for breast neoplasm,breast neoplasm metastasis is still an intractable medical problem.At present,the related mechanisms of breast neoplasm metastasis are as follows:epithelial-mesenchymal transition,dissolution of extracellular matrix,circulating tumor cells or disseminated tumor cells,tumor vessels and tumor microenvironment.Making clear of the related mechanisms of breast neoplasm metastasis has certain guiding significance for the prediction and treatmence of breast neoplasm metastasis.

Objective To optimize the most favorable extraction conditions of Nutmeg. Method With the volatile oils extracting ratio of Nutmeg as optimization targets, the extraction factors affected the volatile oils extracting ratio were optimized by uniform design. Result The optimum condition for extraction was as follows:medicinal materials which screen mesh were 4～10, being soaked in 14 times water for 1 hours and distilled for 6 hours with vapour. Conclusion The extraction process of optimization is simple and practicable, and can provide the test foundation for production.

Objective To investigate the expression and significance of epithelial cell adhesion molecule (Ep-CAM) in breast cancer.Methods Twenty-three cases of breast cancer tissue samples and paired lymph node metastases confirmed pathologically were collected.Isobaric tags for relative and absolute quantitation (iTRAQ) proteomics technology was used to screen and identify the differentially expressed proteins between primary tumor and lymph node metastasis of breast cancer.The expression of Ep-CAM was detected by Western blotting in 4 cases of primary breast cancer tissues and paired lymph node metastases.And the expressions of Ep-CAM in 252 cases of breast lesions were detected by immunohistochemical method.Results Quantitative proteomic examination results showed that differentially expressed proteins existed in breast cancer primary tumor and lymph node metastasis,and the expression of Ep-CAM in metastatic lesions was higher than that in primary tumor.Western blotting results showed that the expression of Ep-CAM in metastatic lesions (1.46 ± 0.22) was higher than that in primary tumor (1.16 ± 0.09),which was consistent with the results of proteomic.The immunohistochemical results showed that the positive expression rate of Ep-CAM in lymph node metastasis tissues (93.16％,109/117) was significantly higher than that in primary breast cancer without metastasis (72.73％,64/88),and the difference was statistically significant (x2 =15.921,P =0.000).The positive expression rate of Ep-CAM in primary breast cancer with lymphatic metastasis (72.65％,85/117) was lower than that in paired lymph node metastases (P =0.001).Conclusion Ep-CAM is differentially expressed in primary tumor and lymph node metastasis of breast cancer,which may be related to the lymphatic metastasis of breast cancer.

Objective To seek differentially expressed proteins for human epithelial growth factorreceptor-2 (HER-2)negative and positive breast carcinoma through establishing proteins profiles,and to providenew prognostic markers and therapeutic targets for patients with breast cancer.Methods HER-2 positiveand negative breast cancer protein expression profiles were established using proteomic isobaric tags for relativeand absolute quantitation (iTRAQ)technology.Differences of protein expression were identified and parts ofdifferential expression proteins were analyzed by bio-informatics,including protein function annotation and GOclassification analysis and Kyoto Encyclopedia of Gene and Genome (KEGG)pathway analysis.Results Proteomicanalysis of breast cancer tissue with identified HER-2 positive and negative groups showed 4 999 differentiallyexpressed proteins by iTRAQ.Based on the criteria of the ratio of HER-2(+)/HER-2(-)≥3,119up-regulated proteins were identified in HER-2 positive group.Based on the criteria of the ratio of HER-2(+)/HER-2(-)≤0.5,47 down-regulated proteins were identified in HER-2 positive group.The results ofGO analysis showed that the molecular function,biological process and cellular composition of differentiallyexpressed proteins were complex between HER-2 positive and negative breast cancer.There were differences inthe distribution of up-regulated proteins and down-regulation of proteins.KEGG pathway analysis showed thatdifferentially expressed proteins involved in 168 signal pathways.Conclusion There are differentiallyexpressed proteins between HER-2 positive and negative breast cancer,which involve complex molecular func-tion,biological process and signaling pathway.

Objective To analyze and determine the clinical, molecular pathology and genetic features of a Chinese family with dystrophinopathy. Methods Clinical data of the proband and his family members were collected. Immunohistochemistry staining was performed on muscular biopsy tissues with antimerosin, emerin and the N, C and central rod domains of dystrophin. Genomic DNA was extracted using standard procedures from the peripheral blood leukocytes. Multiplex ligation-dependent probe amplification (MLPA) was used to test Duchenne muscular dystrophy (DMD) gene to determine the ways and sites of genetic mutation, and analyze the relationships between genotype and phenotype. Results Patients from this family were clinically diagnosed as muscular dystrophy, and they presented serious manifestations although the immunohistochemistry analysis for the proband exhibited partial loss of dystrophin staining, and positive expression with merosin and emerin. Further test with MLPA detected the loss of exons 45--54 in DMD gene in the proband, while his mother had heterozygositic loss in exons 45--54. Conclusions The losses of exons 45--54 in the proband are all derived from his mother, who carries genetic mutation with normal phenotype. He has been diagnosed as dystrophinopathy. At the same time, his partial loss of dystrophin is not parallel to the out-of-frame mutation of the gene and his severe clinical manifestations. Abnormal expression of dystrophin is the pathological basis for dystrophinopathy phenotype. Its clinical outcome depends not only on the degree of the protein expression, but also on the function of the sites where the DMD gene less occurs.

Objective To explore the repair of the open compound wounds in lower extremities caused by multiple factors. Methotis Transplantation of cutaneous.musculo-cutaneous or greater omentum flaps were applied to 155 patients of open compound lower extremity wounds. Results The wound healing rate following first operation was 50% and that following two operations was 14.8%.While the wounds were healed in 7.7% of patients after three operations. Conclusion Transplantations of cutaneous,musculo-cutaneous or greater omentum flaps ale effective to repair and reconstruct the open compound lower extremity wounds.

OBJECTIVETo identify human single chain Fv antibody (ScFv) against hepatitis B viral surface antigen.

METHODSThe recombinant phages were panned by HBsAg which was coated in a microtiter plate, after five rounds of biopanning, 56 phage clones were identified specific to HBsAg. The specificity of ScFv was evaluated by ELISA and immunohistochemistry, respectively.

RESULTSThe data of HB sAg-ScFv DNA digestion and DNA sequencing showed that the ScFv gene is composed of 750 bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis B surface antigen has a specific combination character with hepatitis B surface antigen of different sources and paraffin-embedded patients tissue specimens, it did not react with normal liver tissue and HCV.

CONCLUSIONSThe application of HBsAg specific ScFv in immunohistochemistry was successfully achieved.

Bacteriophages ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Antibodies ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B, Chronic ; diagnosis ; Humans ; Peptide Library