A virus was successfully isolated from a sick cat exhibiting clinical signs such as oral mu-cosal ulceration,nasal mucosal redness,and increased nasal secretions utilizing F81 cells.Through a comprehensive analysis as such PCR amplication,sequencing,morphology,serology,and animal re-gression tests,the virus was identified as a feline calicivirus and named FCV-BJ,an inactivated vac-cine was developed from this isolated strain its safety and efficacy were assessed.The results re-vealed that the isolated FCV-BJ strain exhibited characteristic serological and morphological fea-tures consistent with caliciviruses.Furthermore,inoculation of cats with the FCV-BJ demonstrated the strain is highly virulent and the cats manifested the clinical signs of feline calicivirus infection.For the vaccination trial,domestic cats were immunized with inactivated fifth-generation virus cell culture at varying dilutions,followed by a booster immunization after 21 days.Fourteen days after the challenge with the virus,cats immunized with 107.0 TCID50/mL or higher remained largely healthy,while all cats in the control group developed clinical signs of FCV.These findings suggest that the inactivated vaccine derived from the FCV-BJ isolate exhibits strong immunogenicity and protective efficacy at a minimum immunization dose of 107.0 TCID50/mL.This strain holds promise as a candidate for vaccine production,providing a valuable reference and foundation for future re-search and development of feline calicivirus vaccines.

Breast cancer stem cells (CSCs) are the main causes leading to the failure of treatment of breast cancer and play important roles in the progression of breast cancer and drug resistance, which are closely related to the therapeutic resistance of radiotherapy and chemotherapy, and endocrine therapy.The metastatic potential and therapeutic resistance of CSCs are associated with epithelial mesenchymal transition and Hedgehog, Wnt, interleukin-6/signal transduction and tanscriptional activation factor 3, transforming growth factor-β and other signaling pathways.While some of the targeted drugs targeting these signaling pathways are undergoing clinical transformation, which is expected to provide new approach for the clinical treatment of breast cancer.

OBJECTIVETo study the expression and clinical significance of MTDH and VEGF in triple-negative breast cancer (TNBC).

METHODSTissue samples of 168 breast cancers (including 112 TNBC tissue and 56 non-TNBC tissue), 10 breast fibroadenomas and 15 normal breast tissues were collected. Postoperative specimens were examined by immunohistochemistry for MTDH and VEGF expression. The correlation between the expression of MTDH and VEGF and clinicopathological features was analyzed.

RESULTSMTDH and VEGF were expressed in 57.1% and 49.4% of breast cancer patients, 64.3% and 56.3% in TNBC patients, respectively, significantly higher than that in the non-TNBC tissues, breast fibroadenomas and normal breast tissues (P<0.05 for all). Statistically significant correlation was found between the MTDH and VEGF expressions (r=0.356, P<0.001). Moreover, MTDH expression was correlated with tumor size, BMI index, lymph node metastasis, pathological stage, recurrence and metastasis, and the expression of p53 and Ki-67 proteins (P<0.05 for all). The VEGF protein expression was correlated with lymph node metastasis, pathological staging, recurrence and metastasis, and the expression of Ki-67 protein (P<0.05 for all). The patients with high expression of MTDH and VEGF showed a lower DFS and OS (P<0.05 for both).

CONCLUSIONSMTDH and VEGF expression may be correlated with tumor angiogenesis and progression and has the potential to be valuable prognostic factors in patients with TNBC.

Biomarkers, Tumor ; metabolism ; Breast ; metabolism ; Cell Adhesion Molecules ; metabolism ; Disease-Free Survival ; Female ; Fibroadenoma ; blood supply ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Neoplasm Proteins ; metabolism ; Neovascularization, Pathologic ; Prognosis ; Triple Negative Breast Neoplasms ; blood supply ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effect of mTOR signal transduction pathway and down-regulating anti-oncogene PTEN on the growth of breast cancer MCF-7 cells.

METHODSMCF-7 cells were transfected with the eukaryotic expression plasmid pcDNA3.1-mTOR and non-loaded plasmid, and the expression of mTOR in the cells was detected using Western blotting. Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells, and the expression of PTEN was detected after transfection.

RESULTSThe cells transfected with pcDNA3.1-mTOR showed a increased growth rate than those transfected with the non-loaded plasmid and those without transfection. The expression of the protein PTEN decreased obviously in the cells after mTOR trasnfection.

CONCLUSIONmTOR can regulate the expression of PTEN via PI3K/AKT/PTEN pathways through a negative feedback mechanism. Increased mTOR expression promotes MCF-7 cell growth, suggesting the potential value of mTOR specific inhibitor in the treatment of breast cancer.

Apoptosis ; Breast Neoplasms ; pathology ; Cell Cycle ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; MCF-7 Cells ; PTEN Phosphohydrolase ; metabolism ; Plasmids ; Signal Transduction ; TOR Serine-Threonine Kinases ; genetics ; Transfection

Objective To observe the effect and toxicity of gemcitabine and S-1 in treatment of metastatic triple-negative breast cancer.Methods In this study,41 cases of metastatic breast cancer were treated in the General Hospital of Shenyang Military Region between Jun.2010 and Dec.2012.The median age was 55 years old.The pathological diagnosis of these patients was triple-negative breast cancer.All patients were given gemcitabine 1000 mg/m2 intravenously on the 1st and 8th day,and 60 mg S-1 from the 1st day to the 14th day orally for every cycle.There were 21 days for each cycle.All patients accepted at least 2 cycles of chemotherapy and once effect evaluation.Results 41 cases were diagnosed as metastatic triple-negative breast cancer,with the failure of second-line treatment.The median age was 55 years.All cases were followed up until death.All the 41 cases were administrated for more than 2 cycles,among whom,there were 0 case of complete response(CR),16 cases (39.0％)of partial response(PR),14 cases(34.1％) of stable disease(SD),and 11 cases(26.8％) of progressive disease(PD).The disease control rate was 73.1％ (30/41).In this study,median progression free survival(mPFS)was 7.9 months.The rate of digestive toxicity and marrow suppression was 24.4％ and 55％ respectively.No patient stopped treatment because of severe toxicities.Conclusion The chemotherapy regimen of gemcitabine and S-1 is effective in treatment of metastatic triple-negative breast cancer,and the toxicity could be tolerated.

Objective To investigate the effect of mTOR signal transduction pathway and down-regulating anti-oncogene PTEN on the growth of breast cancer MCF-7 cells. Methods MCF-7 cells were transfected with the eukaryotic expression plasmid pcDNA3.1-mTOR and non-loaded plasmid, and the expression of mTOR in the cells was detected using Western blotting. Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells, and the expression of PTEN was detected after transfection. Results The cells transfected with pcDNA3.1-mTOR showed a increased growth rate than those transfected with the non-loaded plasmid and those without transfection. The expression of the protein PTEN decreased obviously in the cells after mTOR trasnfection. Conclusion mTOR can regulate the expression of PTEN via PI3K/AKT/PTEN pathways through a negative feedback mechanism. Increased mTOR expression promotes MCF-7 cell growth, suggesting the potential value of mTOR specific inhibitor in the treatment of breast cancer.

Objective To investigate the effect of mTOR signal transduction pathway and down-regulating anti-oncogene PTEN on the growth of breast cancer MCF-7 cells. Methods MCF-7 cells were transfected with the eukaryotic expression plasmid pcDNA3.1-mTOR and non-loaded plasmid, and the expression of mTOR in the cells was detected using Western blotting. Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells, and the expression of PTEN was detected after transfection. Results The cells transfected with pcDNA3.1-mTOR showed a increased growth rate than those transfected with the non-loaded plasmid and those without transfection. The expression of the protein PTEN decreased obviously in the cells after mTOR trasnfection. Conclusion mTOR can regulate the expression of PTEN via PI3K/AKT/PTEN pathways through a negative feedback mechanism. Increased mTOR expression promotes MCF-7 cell growth, suggesting the potential value of mTOR specific inhibitor in the treatment of breast cancer.

Objective To investigate if down regulation of MTDH could inhibit proliferation and induce apoptosis in breast cancer SK-BR-3 cells.Methods RNA interference was employed to reduce MTDH expression in human breast cancer SK-BR-3 cells.Western blot assay was applied to measure the down regulation of MTDH.MTT assay was performed to assess the proliferation of SK-BR-3 cell.Flow cytometry was employed to detect cell cycle and apoptosis.Western blot assay was applied to detect the molecular alterations that was associated with cell proliferation,cell cycle and apoptosis.Results MTDH down regulation significantly inhibited cell proliferation.48 hours and 72 hours after trasnfection,the absorbance value(A value)in blank control,negative control and treatment group was (2.0 ± 0.1) vs (1.9 ± 0.3) vs (0.9 ± 0.1) (P =0.02) and (2.7 ± 0.2) vs (2.5 ± 0.4) vs (1.3 ± 0.2) (P =0.008).MTDH down regulation resulted in accumulation of the G0/G1 phase cells and reduction of S and G2/M phase cells.Moreover,MTDH down regulation significantly induced cell apoptosis.The cell apoptosis rate in blank control,negative control and trial group was (1.3 ± 0.2) ％,(1.4 ± 0.3) ％ and (19.6 ± 2.7) ％ (P =0.012).MTDH down regulation resulted in a decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.Conclusions Reduced MTDH expression in SKBR-3 cells can inhibit proliferation and induce apoptosis,which may be associated with decreased expression of cyclinD1 and Bcl-2,an increased expression of P21 and the activation of caspase-3.

Obesity has been identified as one of the risk factors for malignant neoplasms,such as breast cancer.Epidemiological data shows that obesity is closely related to the development and progression of breast cancer.The pathogenesis may involve in estrogen,insulin,leptin,adiponectin and inflammation factors.Therefore,maintain normal body weight may contribute to the prevention of breast cancer.

Prolactin-inducible protein(PIP)is regarded as a kind of specific tumor marker,and detecting the expression of PIP from breast cancer tissues,metastatic lymph nodes and peripheral blood can find breast cancer micro-metastasis effectively.Some technologies such as immunohistochemistry,reverse transcription polymerase chain reaction and so on can detect PIP sensitivily.Recently,these technologies have been used in some clinical and basic studies.Detection rate of breast cancer micro-metastasis is improved effectively when PIP is combined with other tumor makers,especially mammag-lobin and cytokeratin 19.