Globularifolin inhibits CAMA-1 human breast cancer cell line via cell cycle arrest, apoptosis and inhibition of PI3K/AKT signalling pathway

Purpose: To investigate the anticancer activity of globularifolin against CAMA-1 breast cancer cells.Methods: The viability of CAMA-1 cells was assessed by MTT assay. DAPI and annexin V/PI staining were used for the determination of apoptotic cell death. Flow cytometry was employed for cell cycle analysis, while wound healing and immunoblotting assays were used to measure cell migration and protein expression, respectively.Results: Globularifolin decreased the viability of CAMA-1 breast cancer cells dose-dependently, with half-maximal inhibitory concentration (IC50) of 10 μM, relative to its IC50 of 65 μM against non-cancerous CMMT breast cells. It also initiated apoptotic cell death and cell cycle arrest. Moreover, it inhibited the migration of CAMA-1 breast cancer cells, and PI3K/AKT signalling cascade.Conclusion: These results suggest that globularifolin has promising potential for use in the treatment of breast cancer.Keywords: Breast cancer, Globularifolin, Apoptosis, Cell cycle, Cell migratio

A two-dimensional electrophoresis protocol suitable for Medicago truncatula leaf proteome

Medicago truncatula leaves were used as the experimental materials. Total proteins of leaves were extracted by trichloracetic acid (TCA)-acetone method and proteins had a better separation using gel strips, forming an immobilized non-linear 3 to 10 pH gradient focusing 123,000 vhr combined with 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The gels were stained with Coomassie Brilliant Blue G-250 and digitalized gels were analyzed using the PDquest 8.0.1 software. The results indicated that 931 protein dots were detected in the gel. A technology suitable for the M. truncatula leaves protein extraction by TCA/acetone and a protocol for two-dimensional electrophoresis (2-DE) was established, which provides technical support for M. truncatula leaf proteome research.Keywords: Medicago truncatula, proteome, two-dimensional polyacrylamide gel electrophoresis (2-DE), isoelectrofocusing (IEF)African Journal of Biotechnology Vol. 12(3), pp. 233-23

Experimental Study on Dry Friction Damping Characteristics of the Steam Turbine Blade Material with Nonconforming Contacts

An experiment system has been established to study the dry friction damping dynamic characteristics of the steam turbine blade material 1Cr13. The friction dynamic characteristics of the specimens with nonconforming contact surfaces are measured under different parameters. The experiment results are compared with that of the macroslip hysteresis model and the Mindlin microslip friction model in detail. The results show that the experimental result of the tangential contact stiffness is in good agreement with that of the theory result based on the fractal theory and the Hertz contact theory by Jiang et al., 2009. The dimensionless equivalent stiffness and equivalent damping obtained by the macroslip hysteresis model agree well with the experimental results when relative motion is relatively large. However, the results of the macroslip hysteresis model differ a lot from the experimental results when relative motion is relatively small. Compared with the macroslip hysteresis model, the Mindlin microslip friction model can predict the dimensionless equivalent stiffness and equivalent damping accurately during the whole measurement range. The linear regularities of dimensionless equivalent stiffness and equivalent damping are obtained, which decrease the difficulty of building the vibration analysis model of the blade with sufficient accuracy

Mutation Status and Immunohistochemical Correlation of KRAS, NRAS, and BRAF in 260 Chinese Colorectal and Gastric Cancers

KRAS, NRAS and BRAF are kinases involved in the RAS-RAF-MAPK signaling pathway and also potential tumor-driven genes. Patients with KRAS/NRAS/BRAF mutations are resistant to anti-EGFR monoclonal antibody therapy. The main purpose of this study is to investigate the mutation status and distribution of KRAS/NRAS/BRAF in Chinese colorectal and gastric cancers, and to explore the histopathological changes and related immunohistochemical marker changes caused by these mutations. The mutation status of KRAS (exons 2, codon 12/13), NRAS (exons 2/3/4, codon 12/13/59/61/117/146) and BRAF (exons 15, codon 600) were detected by amplification refractory mutation system polymerase chain reaction (ARMS-PCR) in 86 colon cancer, 140 rectal cancer and 34 gastric cancer tissues. Then, the frequencies and distribution of KRAS/NRAS/BRAF mutations were described in detail. Furthermore, the relationship between KRAS/NRAS/BRAF mutations and the features of histopathological and related immunohistochemical markers were analyzed. The results showed that KRAS/NRAS/BRAF mutation rates in colon cancer were 44.2, 1.2, and 3.5%; in rectal cancer were 37.1, 4.3, and 0.7%; in gastric cancer were none, none and 2.9%. The mutation rate of KRAS in female (48.8%) is significantly higher than that of male (27.8%), and the mutation rate increased with the higher degree of differentiation. Additionally, the mutation rate of BRAF detected by ARMS-PCR (1.77%) was significantly lower than that by immunohistochemistry (4.11%). It also showed that the KRAS/NRAS/BRAF mutation status had a certain relationship with the expression of some immunohistochemical markers. This study provides more data support for clinical research on KRAS/NRAS/BRAF mutation in CRCs or gastric cancers