ABSTRACT
Objective To explore the application of erlotinib based targeting fluorescent probe in the detection of lung cancer.Methods The erlotinib based targeting fluorescent probe was prepared.The lung cancer cells of A-549 were selected as experimental group,and cervical cancer cells of CaSki,SiHa and C33-A were selected as control group.The A-549,CaSki,SiHa and C33-A cells were identified by 0.1 × 10-6,1 × 10-6,10 × 10-6 mol · L-1 fluorescent probe;the identification ability of erlotinib based targeting fluorescent probe on cells in the two groups was observed under the inverted fluorescence microscope.Results When the concentration of fluorescent probe was 10 × 10-6 mol · L-1,the strong fluorescence signal was observed in A-549 and CaSki cells,but the fluorescence signal was not observed almost in SiHa and C33-A cells.When the concentration of fluorescent probe was 1 × 10-6 mol · L-1,the strong fluorescence signal was observed in A-549 cells;the weak fluorescence signal was observed in C33A cells;the fluorescence signal was not observed almost in SiHa and C33-A cells.When the concentration of fluorescent probe was 0.1 × 10-6 mol · L-1,the strong fluorescence signal was observed in A-549 cells;the fluorescence signal was not observed almost in CaSki,SiHa and C33-A cells.Conclusion Erlotinib based targeting fluorescent probe can specifically recognize lung cancer A-549 cells.
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Objective To clarify which adenosine receptor subtype is the most powerful one on controlling retinal pigment epithelial cell (RPE) binding adenosine, and what is its function in RPE. Methods Total mRNA was isolated, and membrane protein was extracted from in vitro cultured human ARPE-19 cells. For all four kinds of adenosine receptors, ARA1, ARA2A, ARA2B and ARA3, their gene expressions were tested by real-time PCR while their molecules in the membrane protein were detected by Western blot assay. To check the influence of each adenosine receptor subtype on ARPE-19 cell binging ability to adenosine the cultured cells were divided into five groups, named A-E. A group was set up as untreated control, while, groups B-E were separately treated by ARA1 agonist DPCPX (50 nmol/L), ARA2A agonist SCH58261 (100 nmol/L), ARA2B agonist MRS1754 (100 nmol/L) or ARA3 agonist MRS1220 (5μmol/L). H3-adenosine a radioactive ligand binding assay was performed and the maximum binding capacities (Bmax) were calculated in groups A-E of ARPE-19 cells. Then, ARPE-19 cells were all treated by the combination of TNF-αand IFN-γbut with or without CCPA (100 nmol/L), an ARA1 agonist. MCP-1, IP-10, IL-6, IL-10 and TGF-β in their mediums were determined by ELISA. Results Either mRNA expression or membrane localization of ARA1, ARA2A, ARA2B and ARA3 were verified by real-time PCR and Western blot assay respectively. For A-E groups of ARPE-19 cells the Bmax of adenosine binding were (2.04± 0.31), (0.44 ± 0.06), (1.82 ± 0.28), (2.01 ± 0.42) and (2.06 ± 0.44) fmol respectively;and which were statistically decreased in group B than those of all other groups (P<0.01). Compared with control RPE, the contents of IL-6, MCP-1 and IP-10 were decreased after treatment with CCPA, and the content of IL-10 increased in RPE group (P<0.01). There was no significant difference in TGF-β content between the two groups. Conclusion APRE-19 cells predominantly use ARA1 to absorb adenosine, and the activation of ARA1 in ARPE-19 cells inhibits its IL-6, MCP-1, and IP-10 production, which have potentially immunosuppressive effects to APRE-19 cells.
ABSTRACT
Due to the low effectiveness of traditional assisted reproductive technology (ART), new technological possibilities are constantly explored. Lots of studies have demonstrated the potential of microfluidics to revolutionize the fundamental processes of in vitro fertilization (IVF). With the advantages of high efficiency, short time, harmless collection, real-time observation of separation, similar microenvironment, and automation, the application of microfluidics in sperm isolation and IVF has shown an evident superiority over the conventional approaches and provided a new platform for ART. This review highlights the application of various microfluidic techniques in sperm motility assessment and isolation, sperm chemotaxis assay, IVF, sperm concentration, and sperm separation and enrichment in recent years. It also briefly introduces the basic principles, structural design, and operation processes of the microfluidic platform, focusing on the advantages and disadvantages of each method and the potential of their clinical application. Obviously, there are still some challenges to the application of microfluidics in ART. However, it is believed that the development of this new technology would be toward a highly integrated application of several steps in one single device, known as IVF-lab-on-a-chip.