A high-throughput study on endothelial cell adhesion and growth mediated by adsorbed serum protein via signaling pathway PCR array

The purpose of this paper is to utilize the signaling pathway polymerase chain reaction(PCR)arrays to investigate the activation of two important biological signaling pathways in endothelial cell adhesion and growth mediated by adsorbed serum protein on the surface of bare and titanium nitride(TiN)-coated nickel titanium(NiTi)alloys.First,the endothelial cells were cultured on the bare and TiN-coated NiTi alloys and chitosan films as control for 4 h and 24 h,respectively.Then,the total RNA of the cells was collected and the PCR arrays were performed.After that,the differentially expressed genes in the transforming growth factor beta(TGF-b)signaling pathway and the regulation of actin cytoskeleton pathway were screened out;and the further bioinformatics analyses were performed.The results showed that both TGF-b signaling pathway and regulation of actin cytoskeleton pathway were activated in the cells after 4 h and 24 h culturing on the surface of bare and TiN-coated NiTi alloys compared to the chitosan group.The activated TGF-b signaling pathway promoted cell adhesion;the activated regulation of actin cytoskeleton pathway promoted cell adhesion,spreading,growth and motility.In addition,the activation of both pathways was much stronger in the cells cultured for 24 h versus 4 h,which indicated that cell adhesion and growth became more favorable with longer time on the surface of two NiTi alloy materials.

A New High-throughput Real-time PCR Assay for the Screening of Multiple Antimicrobial Resistance Genes in Broiler Fecal Samples from China

Objective Antimicrobial resistance(AMR)has become a global concern and is especially severe in China.To effectively and reliably provide AMR data,we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology,and screened multiple AMR genes in broiler fecal samples.Methods A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection.A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR genes.Results The new assay with limits of detection ranging from 40.9 to 8,000 copies/reaction.The sensitivity rate,specificity rate,positive predictive value,negative predictive value and correct indices were 99.30%,98.08%,95.31%,99.79%,and 0.9755,respectively.Utilizing this assay,we demonstrate that AMR genes are widely spread,with positive detection rates ranging from 0 to 97.07%in 273 broiler fecal samples.bla CTX-M,bla TEM,mcr-1,fex A,cfr,optr A,and int I1 showed over 80%prevalence.The dissemination of AMR genes was distinct between the two farms.Conclusions We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal samples.The widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.

Viscum Album Modulates Apoptotic Related Genes in Melanoma Tumor of Mice

Cancer is a major public health problem throughout the world. It is estimated that one third of the American population will develop the disease at some time during their lifetimes. Among these, melanoma will account for 7% of the cases. In Brazil, in 2012, it is estimated that over six thousand new melanoma cases occurred. During recent years, the incidence of melanoma has increased, mainly due to a more constant exposure of the skin to sunlight. In this work, our aim is to assess the expression of apoptotis-related genes melanoma tumors in mice treated with Viscum album (VA). This will allow us to better understand the molecular mechanisms underlying tumor cell death activation caused by this compound. Our results clearly demonstrate upregulation of pro apoptotic genes (Trp53bp2, Nol3, Fadd, Tnfsf10, Traf1, Traf2, Cflar, Card10, Nod1, Casp 2, Casp7, Xiap, Dad1, and Dffb). Further bioinformatics-based tool analysis allowed us to assess which specific cell death-related intracellular pathways were activated by VA treatment. Two major effects of VA in melanoma cells could be observed: generation of an immunomudulatory Th-1 like action, recruiting several interleukines, and cell death activation through Caspase7, associated uspstream with Card10 and downstream with CAD. In summary, VA modulates apoptosis related genes in cancer melanoma cells. Although a deeper study should be conducted, VA seems to interfere with important signaling pathways within melanoma cells that control the cellular mechanisms of apoptosis activation. Therapeutic approaches using VA as an antineoplastic and adjuvant medication compounding should be considered.

血清miRNA表达谱在肝细胞癌中的差异性研究

[目的]探讨肝细胞癌、肝硬化患者与正常人血清miRNA表达水平的差异性;筛选出肝癌中差异较明显的血清miRNA谱。[方法]分别收集4例肝癌患者、3例肝硬化患者和3例同期正常体检人员的血清,采用Qiagen miRNA PCR Array芯片法筛查肝癌、肝硬化患者和正常人差异表达的血清miRNA。[结果]初步筛查出了差异表达的血清miRNA,与正常人比较,以相差2倍为分界,肝癌血清miRNA共128个下调,98个上调;肝硬化血清miRNA共131个下调,74个上调;其中,肝癌和肝硬化患者血清miRNA同时下调的有76个,同时上调的有53个。与肝硬化比较,以相差2倍为分界,肝癌血清miRNA共108个下调,103个上调;结合文献、P值及CT值进一步筛选出差异最有意义的7个与肝癌相关的血清miRNA(miR-373-5p,miR-1290,miR-4516,miR-134-5p,miR-144-5p,miR-125a-5p,miR-126-3p)。[结论]差异表达的miRNA谱与肝癌的发生、发展有一定的相关性。

miR-34a: Multiple Opposing Targets and One Destiny in Hepatocellular Carcinoma

Background and Aims:The role of miR-34a in hepatocellular carcinoma (HCC) is controversial and several unresolved issues remain,including its expression pattern and relevance to tumor etiology,tumor stage and prognosis,and finally,its impact on apoptosis.Methods:miR-34a expression was assessed in hepatitis C virus (HCV)-induced non-metastatic HCC tissues by RT-Q-PCR.Huh-7 cells were transfected with miR-34a mimics and the impact of miR-34a was examined on 84 pro-apoptotic/anti-apoptotic genes using PCR array;its net effect was tested on cell viability via MTTassay.Results:miR-34a expression was up-regulated in HCC tissues.Moreover,miR-34a induced a large set of pro-apoptotic/anti-apoptotic genes,with a net result of triggering apoptosis and repressing cell viability.Conclusions:HCC-related differential expression of miR-34a could be etiology-based or stage-specific,and low expression of miR-34a may predict poor prognosis.This study's findings also emphasize the role of miR-34a in apoptosis.