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Objective:To discuss the influence of ALDH1+and CD133+phenotypic breast cancer stem-like cells in TA2 triple negative breast cancer on promoting epithelial-mesenchymal transition (EMT) occurrence in TA2 mice with triple-negative breast cancer and on their biological behavior. Methods:Flow cytometry was performed to analyze the markers ALDH1 and CD133 in TA2 mice triple nega-tive breast cancer and breast cancer stem-like cells with ALDH1+, ALDH1?, CD133+, and CD133?phenotypes, which were sorted out. Then, the TA2 mice were inoculated with sorted tumor cells according to cell type. The mice were divided into ALDH1+, ALDH1?, CD133+, and CD133-groups. The tumor-growing conditions were observed. A tumor tissue was sliced for the immunohistochemical testing of ALDH1?, CD133?, and EMT-related Twist1, E-cadherin, and VE-cadherin proteins. The expression difference of breast cancer stem cell surface markers ALDH1 and CD133 in triple-negative breast cancer and EMT-related proteins Twist1, E-cadherin, and VE-cad-herin was analyzed. Results:The expression rates of breast cancer stem cell markers ALDH1 and CD133 in TA2 mice triple negative breast cancer were 31.2%and 6.5%, respectively. The tumor growth ability of TA2 mice from ALDH1+group was obviously stronger than that from ALDH1?group. The CD133+group was evidently stronger than CD133?group. The immunohistochemical results showed that ALDH1, Twist1, and VE-cadherin expression levels in the ALDH1+group were evidently higher than that in the ALDH1?group (all P<0.05). E-cadherin expression decreased (P<0.05). CD133?, Twist1, and VE-cadherin expression levels in CD133+group were higher than that in CD133?group (all P<0.05). Conclusion:In TA2 mice triple negative breast cancer, ALDH1+and CD133+phenotypic breast cancer stem-like cells may influence the expression of EMT-related proteins, and promote the formation of triple-negative breast cancer.
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Objective To construct and screen the human immunodeficiency virus-1 (HIV-1) negative regulation factor (Nef) peptide-specific CD4+ T lymphocyte clone.Methods Peripheral blood mononuclear cells (PBMC) from five asymptomatic HIV-1 infected patients were collected and Bulkcultured with Nef end peptides.The CD4 molecule and intracellular interferon (IFN)-gamma of cultured cells were detected by two-color flow cytometry.The Nef end peptide-specific T cell clone was then constructed by limited dilution and confirmed through enzyme linked immunospot assay (ELISPOT).The best grown cells were selected and cultured as the final clone.Results The Nef end peptide-specific-T lymphocyte clone was successfully constructed from PBMC of one HIV-infected patient and confirmed by ELISPOT.The detection of human leukocyte antigen (HLA)-DRB1 type showed that the epitope of this peptide was probably HLA-DRB1 * 0406.Conclusion The Nef end specific-T cell clone is successfully constructed,and a new epitope in the C-terminus of Nef protein and its HLA restriction are identified.
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Objective To systematically evaluate the clinical efficacy of Lanqin Oral Liquid(LOL)and ribavirin on infantile hand-foot-mouth disease(HFMD).Methods The randomized controlled trials(RCT)of LOL and ribavirincombined for treating HFMD were retrieved from EMbase, PubMed, CNKI, CBM and VIP(from establish to Oct 2014), the methodological quality of included literature was evaluated, and data analyses were performed with RevMan 5.2 software.Results Fifteen studies involving 2 016 patients were ultimately identified.The meta-analysis results showed that the effective rate of LOL group was superior to that of the control,OR(95%CI)was 5.80 (3.94, 8.52), and there were significant differences in the antifebrile time, erythra regression time, herpes regression time and oral ulcer regression time between the groups,MD (95%CI): -1.29 (-1.45,-1.13), -1.88 (-2.44,-1.33), -1.57 (-2.36,-0.78), -1.57 (-2.07,-1.08), respectively. Conclusion Based on the present clinical evidence, the meta-analysis results indicate that LOL and ribavirin treating HFMD is effective and safe. However, due to the uneven quality of these included studies, this conclusion need more large sample size and high-quality clinical RCTs to be confirmed.
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Objective:To investigate the clinical significance of epithelial-to-mesenchymal (EMT) in lung squamous cell carcino-ma (LSCC) and to examine the effect of EMT on the invasive and migration abilities of LSCC. Methods:Immunohistochemical stain-ing was performed to determine the expression of E-cadherin, Vimentin, and TGF-β1 in 79 LSCC patients, and the clinical significance was explored. SK-MES-1 lung squamous carcinoma cells were cultured in conditioned medium containing various concentrations of transforming growth factor-β1 (TGF-β1) for 5 and 10 days. The expression levels of E-cadherin and Vimentin were detected via West-ern blot and reverse transcription-polymerase chain reaction (RT-PCR). With different concentrations and induction times, invasion and wound healing assays were performed to evaluate the invasion and migration abilities. Results:E-cadherin expression was significantly lower, whereas Vimentin expression was significantly higher in LSCC with lymph node metastasis than in that without noda metastasis (P<0.05). In the tissues of 79 LSCC patients, TGF-β1 expression was significantly related to lymph node metastasis (P<0.05). Western blot showed that Vimentin expression was higher, whereas E-cadherin expression was lower in TGF-β1 inducing medium with 10 ng/mL SK-MES-1 cells than in the other media. RT-PCR showed similar results. Scratch test and invasion assay both showed that treat-ment of cells with cytokines markedly enhanced the migration and invasion of the cells. Conclusion:Lymph node metastasis of LSCC correlates with EMT. SK-MES-1 cells undergo EMT via TGF-β1 induction, which enhances invasion and migration.
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Since there is a clinical need for the tissue-engineered vascular graft (TEVG), fabricating the vascular scaffold individually appears to be necessary. In this work, we have developed the traditional tubular scaffold and branch vascular scaffold utilizing low-temperature deposition manufacturing (LDM) technology. Then different tubular scaffolds were fabricated by changing the processing parameters, and the morphological properties of the scaffolds were assessed. The scaffolds reproduced the structure of 3D vascular model accurately. Wall thickness of the scaffold increased with the increase of velocity ratio (V(L)/V(s)) and nozzle temperature, and both the micropore size and wall roughness were positively correlated with the nozzle temperature. However, the porosity was barely affected by the nozzle temperature. This approach, fabricating vascular scaffold with special structure and appearance features via LDM technology, is potential for the individual fabrication of vascular scaffold.
Subject(s)
Humans , Biocompatible Materials , Chemistry , Blood Vessel Prosthesis , Cold Temperature , Computer-Aided Design , Lactic Acid , Chemistry , Polyesters , Polymers , Chemistry , Tissue Engineering , Methods , Tissue ScaffoldsABSTRACT
Three-dimensionally controlled cell-assembly technique makes fabricating tissues and organs in vitro to be possible. However, for real tissues and organs with complex structure and various cells, fabricating tissues and organs in vitro need a technique that could assemble and locate multi cells and materials precisely in the space. Facing the needs of multi-cell assembly, we designed a mixer nozzle and the matching pulse switching circuit which based on the single-nozzle cell assembly system, and developed a multi-cell-assembly system. We also carried out some assembly experiments with this system using materials that were similar to the multi-component extracellular matrix materials. The results demonstrated that the system could assemble various cells and materials into three-dimensional inhomogeneous structures precisely.