Epithelial ovarian cancer is the most lethal gynecological cancer mainly due to late diagnosis, easy spreading and rapid development of chemoresistance. Cancer stem cells are considered to be one of the main mechanisms for chemoresistance, as well as metastasis and recurrent disease. To explore the stemness characteristics of ovarian cancer stem cells, we successfully enriched ovarian cancer stem-like cells from an established ovarian cancer cell line (SKOV-I6) and a fresh ovarian tumor-derived cell line (OVS1). These ovarian cancer stem-like cells possess important cancer stemness characteristics including sphere-forming and self-renewing abilities, expressing important ovarian cancer stem cell and epithelial–mesenchymal transition markers, as well as increased drug resistance and potent tumorigenicity. Microarray analysis of OVS1-derived sphere cells revealed increased expression of amphiregulin (AREG) and decreased expression of its conserved regulatory microRNA, miR-34c-5p, when compared with the OVS1 parental cells. Overexpression of AREG and decreased miR-34c-5p expression in SKOV-I6 and OVS1 sphere cells were confirmed by quantitative real-time PCR analysis. Luciferase reporter assay and mutant analysis confirmed that AREG is a direct target of miR-34c-5p. Furthermore, AREG-mediated increase of sphere formation, drug resistance toward docetaxel and carboplatin, as well as tumorigenicity of SKOV-I6 and OVS1 cells could be abrogated by miR-34c-5p. We further demonstrated that miR-34c-5p inhibited ovarian cancer stemness through downregulation of the AREG-EGFR-ERK pathway. Overexpression of AREG was found to be correlated with advanced ovarian cancer stages and poor prognosis. Taken together, our data suggest that AREG promotes ovarian cancer stemness and drug resistance via the AREG-EGFR-ERK pathway and this is inhibited by miR-34c-5p. Targeting AREG, miR-34c-5p could be a potential strategy for anti-cancer-stem cell therapy in ovarian cancer.

This case report describes an unusual granulomatous lesion that was curetted from the apices of 2 endodontically treated teeth. Microscopic evaluation revealed a radicular cyst with irregularly shaped eosinophilic hyaline bodies in close association with giant cells. Sections were stained with periodic acid‐Schiff with and without diastase to evaluate the nature of the hyaline bodies. These stains revealed the presence of a carbohydrate material, possibly of plant origin. However, since starch cells could not be demonstrated morphologically, a definitive diagnosis of pulse granuloma was not made.

Summary. -Outbreaks of highly pathogenic avian influenza have caused considerable economic losses in the poultry industry and have also resulted in human deaths since 2004. Rapid subtyping of highly pathogenic avian influenza viruses(HPAIVs) in clinical specimens is a prerequisite of prompt control of disease and prevention of its spreading. In this study, we describe development of a DNA microarray-based detection and subtyping of HPAIVs in field samples. DNA copies of matrix (M) protein genes for the H5, H7, and H9 subtypes of hemagglutinin (HA) and the N1 and N2 subtypes of neuraminidase (NA) were prepared by RT-PCR and specific primers and then spotted onto aldehyde slides to form DNA microarrays. The HPAIV samples to be tested were subjected to total RNA isolation, RT-PCR with universal primers and Cy3 labeling, and the obtained double-stranded DNAs (targets) were finally hybridized with DNA microarrays (probes). A fluorescent spot on the microarray, detected by scanning indicated positive hybridization, i.e. the involved subtype. The assay was specific as various heterologous viruses or HPAIVs of other subtypes tested were negative. No cross-hybridization among different subtypes could be detected. The assay was more sensitive than RT-PCR and chicken embryo inoculation and could be also used for field samples. Summing up, the assay has proved useful for simultaneous detection and differentiation of main epidemic HPAIV subtypes.

BackgroundPanax notoginseng saponins (PNS), the main active ingredient extracted from the Chinese herb SanQi, has antiapoptotic, antioxidant, and anti-inflammatory effects to protect the heart from myocardial ischemia/reperfusion injury (MIRI). ATP-sensitive potassium (KATP) channel plays an important role in pharmacological preconditioning against MIRI. However, its effect on inflammation induced by MIRI remains elusive. Thus, we hypothesise that PNS alleviates the inflammatory response induced by MIRI, and KATP channels play a crucial role in it.MethodsThe rat MIRI model was established by 40 min left anterior descending coronary artery occlusion followed by 60 min reperfusion. PNS was administered 30 min before ischemia, glibenclamide was administered 10 min before PNS. Cardiac function, myocardial histology, and MPO activity were evaluated. The levels of inflammatory mediators TNF-α and MCP-1 in myocardium were measured by ELISA. The protein and gene expression of KATP channel subunits were determined by Western blot analysis and real-time PCR.ResultsPNS pretreatment ameliorated cardiac function and myocardial pathological changes, decreased myocardial MPO activity and inhibited the gene and protein expression of TNF-α and MCP-1. Importantly, PNS increased the expression of the KATP channel. The non-selective KATP channel blocker glibenclamide could block the cardioprotection of PNS.ConclusionPNS preconditioning may reduce inflammation to due to MIRI via KATP channels.AcknowledgementsSupported by a project grant from National Natural Science Foundation (NSFC) of China (Grant No. 81303256).

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