Circular circ_0000885 promotes hepatocellular carcinoma proliferation by epigenetically upregulating Caprin1.

OBJECTIVE: Recently, the vital role of circular RNAs is discovered in many diseases, including tumor progression. Hepatocellular carcinoma (HCC) is one of the most ordinary malignant tumors. The purpose of our study is to detect the potential function of circ_0000885 in HCC to offer new biomarkers and targets. PATIENTS AND METHODS: The expression level of circ_0000885 in HCC tissues and cell lines was monitored by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Pearson's Chi-square test was used to determine the association of circ_0000885 expression with several clinicopathological factors. Then knockdown of circ_0000885 was constructed to uncover its function in HCC. The cell growth ability was measured through the cell counting kit-8 (CCK-8) assay, colony formation assay, and cell cycle assay. The Western blot assay was performed to analyze the protein level of Caprin1. RESULTS: Circ_0000885 was highly expressed in HCC tissues than that in adjacent samples. The miR-532-5p expression was associated with lymphatic metastasis and TNM stage. The expression of circ_0000885 was also higher in HCC cell lines. The cell growth ability of HCC cells was inhibited after circ_0000885 was silenced. Furthermore, Caprin1 was inhibited via knockdown of circ_0000885. CONCLUSIONS: Circ_0000885 could enhance cell proliferation and regulate cell cycle of HCC by promoting Caprin1.

Preliminary results indicate resveratrol affects proliferation and apoptosis of leukemia cells by regulating PTEN/PI3K/AKT pathway.

OBJECTIVE: PTEN-PI3K/AKT signaling pathway is widely involved in the regulation of cell proliferation, cell cycle, apoptosis, and invasion. Resveratrol (Resv) is a natural botanical ingredient involved in several biological activities. It is still unclear in terms of whether Resv may exert anti-leukemia effects by regulating the PTEN-PI3K/AKT pathway. This study investigated the effect of Resv on leukemia cell proliferation and apoptosis by regulating PTEN-PI3K/AKT pathway. PATIENTS AND METHODS: Human normal peripheral blood PBMC cells, and human acute promyelocytic leukemia (APL) cell line NB-4 and HL-60 cells were cultured in vitro. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect Phosphatase and tensin homolog (PTEN) mRNA expression. Western blot was adopted to test PTEN protein expression. HL-60 and NB-4 cells were treated with 0, 5, 10, and 20 µM Resv, respectively. Cell proliferation was analyzed by cell counting kit8 (CCK-8) assay. The level of caspase-3 was measured by Western blot. HL-60 cells were divided into control group, 20 µM Resv treatment group, and Resv+PTEN inhibitor SF1670 group. Cell apoptosis was determined by flow cytometry. Cell proliferation was assessed by EdU staining. RESULTS: Compared with peripheral blood mononuclear cell (PBMC), PTEN mRNA and protein levels were significantly decreased in NB-4 and HL-60 cells. Resv significantly inhibited the proliferation activity in HL-60 and NB-4 cells, and increased the activity of caspase-3. Resv treatment up-regulated the expression of PTEN and reduced the expression of p-AKT protein in HL-60 cells. However, Resv treatment markedly suppressed the proliferation of HL-60 and induced apoptosis. SF1670 treatment in the presence of Resv significantly antagonized the down-regulation of p-AKT protein expression induced by Resv, resulting in decreased apoptosis and enhanced cell proliferation. CONCLUSIONS: Resv can up-regulate PTEN expression and inhibit the activity of PI3K/AKT pathway to play an anti-leukemia effect through suppressing cell proliferation and inducing apoptosis.

Inhibition of the spindle assembly checkpoint kinase TTK enhances the efficacy of docetaxel in a triple-negative breast cancer model.

BACKGROUND: Triple-negative breast cancers (TNBC) are considered the most aggressive type of breast cancer, for which no targeted therapy exists at the moment. These tumors are characterized by having a high degree of chromosome instability and often overexpress the spindle assembly checkpoint kinase TTK. To explore the potential of TTK inhibition as a targeted therapy in TNBC, we developed a highly potent and selective small molecule inhibitor of TTK, NTRC 0066-0. RESULTS AND CONCLUSIONS: The compound is characterized by long residence time on the target and inhibits the proliferation of a wide variety of human cancer cell lines with potency in the same range as marketed cytotoxic agents. In cell lines and in mice, NTRC 0066-0 inhibits the phosphorylation of a TTK substrate and induces chromosome missegregation. NTRC 0066-0 inhibits tumor growth in MDA-MB-231 xenografts as a single agent after oral application. To address the effect of the inhibitor in breast cancer, we used a well-defined mouse model that spontaneously develops breast tumors that share key morphologic and molecular features with human TNBC. Our studies show that combination of NTRC 0066-0 with a therapeutic dose of docetaxel resulted in doubling of mouse survival and extended tumor remission, without toxicity. Furthermore, we observed that treatment efficacy is only achieved upon co-administration of the two compounds, which suggests a synergistic in vivo effect. Therefore, we propose TTK inhibition as a novel therapeutic target for neoadjuvant therapy in TNBC.

Modulation of dendritic cell function by the radiation-mediated secretory protein γ-synuclein.

Recently, γ-synuclein (SNCG), which is also known as breast cancer-specific gene-1, has been demonstrated to be an adverse and aggressive marker in breast cancer. In our previous study, SNCG was significantly upregulated in irradiated human breast cancer cells. The aim of this study was to investigate whether radiation-induced, tumor-derived SNCG can influence dendritic cell (DC) function in immune systems. The phenotypical and functional changes of DCs in the presence or absence of SNCG were investigated by FACS analysis, ELISA, and real-time PCR. The ability of SNCG-treated DCs to influence T cells was also examined by coculturing with T cells. The treatment of DCs with SNCG protein inhibited the surface expression of the co-stimulatory molecules CD40 and CD86, and decreased the mRNA levels of pro-inflammatory cytokines. The SNCG-treated DCs inhibited T-cell proliferation slightly, but distinctively increased the population of regulatory T cells. In addition, the production of TGF-ß from T cells was significantly increased when they were cocultured with SNCG-treated DCs. Taken together, these results demonstrate that tumor-derived SNCG contributes to immunosuppressive effects via the inhibition of DC differentiation and activation, thus making it a potential target for cancer treatment.

Myocardial protection in donor heart preservation: a comparison between Bretschneider's histidine-tryptophan-ketoglutarate solution and cold blood cardioplegia.

BACKGROUND: Optimal myocardial protection for donated hearts is crucial to improve outcomes of heart transplantation and reduce morbidity and mortality. This study aimed to compare the efficacy of myocardial protection using single dose of Bretschneider's histidine-tryptophan-ketoglutarate (HTK) solution and repeated doses of cold blood cardioplegia (CBC) in donor heart preservation. METHODS: Sixty-seven patients undergoing heart transplantation in Tri-Service General Hospital, Taipei, Taiwan between 2002 and 2012 were enrolled in this study. Patients were divided into an HTK group and a CBC group based on the preservation solution used to protect the donated hearts. The perioperative variables and postoperative outcomes were retrospectively reviewed. RESULTS: There were no statistic differences about demographic data in donors and recipients between the 2 groups. There were no significant differences in postoperative cardiac enzymes, hemodynamic data, length of stay in intensive care, or 30-day mortality between the groups. The HTK group showed a trend of shorter pumping time (P = .091). Multivariate analyses reveal that the HTK group had higher postoperative inotropic score (P < .001) and shorter pumping time (P = .02). CONCLUSIONS: Single dose of Bretschneider's HTK solution could effectively reduce pumping time and afford similar myocardial protection compared with repeated doses of CBC in the preservation of donated hearts.

Wip1 suppresses apoptotic cell death through direct dephosphorylation of BAX in response to γ-radiation.

Wild-type p53-induced phosphatase 1 (Wip1) is a p53-inducible serine/threonine phosphatase that switches off DNA damage checkpoint responses by the dephosphorylation of certain proteins (i.e. p38 mitogen-activated protein kinase, p53, checkpoint kinase 1, checkpoint kinase 2, and uracil DNA glycosylase) involved in DNA repair and the cell cycle checkpoint. Emerging data indicate that Wip1 is amplified or overexpressed in various human tumors, and its detection implies a poor prognosis. In this study, we show that Wip1 interacts with and dephosphorylates BAX to suppress BAX-mediated apoptosis in response to γ-irradiation in prostate cancer cells. Radiation-resistant LNCaP cells showed dramatic increases in Wip1 levels and impaired BAX movement to the mitochondria after γ-irradiation, and these effects were reverted by a Wip1 inhibitor. These results show that Wip1 directly interacts with and dephosphorylates BAX. Dephosphorylation occurs at threonines 172, 174 and 186, and BAX proteins with mutations at these sites fail to translocate efficiently to the mitochondria following cellular γ-irradiation. Overexpression of Wip1 and BAX, but not phosphatase-dead Wip1, in BAX-deficient cells strongly reduces apoptosis. Our results suggest that BAX dephosphorylation of Wip1 phosphatase is an important regulator of resistance to anticancer therapy. This study is the first to report the downregulation of BAX activity by a protein phosphatase.

Analyses of non-synonymous obesity risk alleles in SH2B1 (rs7498665) and APOB48R (rs180743) in obese children and adolescents undergoing a 1-year lifestyle intervention.

Association of obesity risk alleles of single nucleotide polymorphisms (SNPs) near or in the SH2B adaptor protein 1 gene (SH2B1) and increased body mass index (BMI) has been often described. A gene in close proximity, apolipoprotein B48 receptor gene (APOB48R), is tagged by the same SNP(s).We analyzed 454 overweight and obese children and adolescents (10.8±2.6 years, BMI-SDS 2.4±0.5; 55% girls) who completed a 1-year lifestyle intervention ('Obeldicks' program). Carriers of obesity risk alleles of non-synonymous SNPs in SH2B1 (rs7498665, Thr484Ala) or APOB48R (rs180743, Pro419Ala), as genotyped by TaqMan, were analysed for changes in anthropometrics (body-mass index (BMI), and standardized BMI (BMI-SDS)), blood pressure (systolic and diastolic) and plasma parameters (total cholesterol, LDL-cholesterol, HDL-cholesterol, triacylglycerides, glucose, insulin, and HOMA).We observed no evidence for an association of the obesity risk alleles to alterations in any of the analyzed phenotypes. Both mean BMI and BMI-SDS improved during the intervention independent of genotype. The mean systolic blood pressure was lowered and concentrations of HDL-cholesterol increased significantly.The obesity risk alleles of non-synonymous SNPs at SH2B1 and APOB48R have no strong effect on weight loss-related phenotypes in overweight children after a 1-year lifestyle intervention.

Prevalence of classical swine fever virus in domestic pigs in South Korea: 1999-2011.

The major policy for eradication of classical swine fever (CSF) in South Korea has focused on the implementation of compulsory vaccination of the susceptible pig population. A vaccine strain of CSF virus, the LOM strain, is used to maintain high herd seroconversion, a practice complementary to the 'stamping-out policy' and restriction of animal movement during disease outbreaks. To survey for the prevalence of CSF in domestic pigs in South Korea over the past 13 years (1999-2011), we tested 4 193 782 and 1 162 645 samples for antibodies and antigens, respectively. Whereas seropositivity for CSF antibodies has been maintained at over 95% in the mainland, in Jeju Island, where no-vaccination has been administered since 1999, seroprevalence has been below 1% during the last 3 years of study (2009-2011). The highest number of outbreaks in South Korea occurred in 2002 and 2003; since then, outbreaks have decreased each year, with the last CSF outbreak recorded in 2009. No outbreaks have occurred during the past 3 years, and a high level of herd immunity has been maintained in the mainland pig population for 8 years; therefore, South Korea could now switch to a no-vaccination policy throughout the country. However, the constant threat of the re-emergence of the disease in the susceptible pig population should be the main consideration in planning and carrying out the last phase of the CSF eradication process.

Multiple nodular fasciitis in the mandibular border area which is misdiagnosed as metastatic lymph node.

Nodular fasciitis (NF) is a benign lesion that has proliferative fibroblasts and myofibroblasts. NF is similar to a tumour and has infiltrative properties. We describe a rare case of multiple nodular fasciitis occurring in the mandibular border area of a 51-year-old male. Radiological and histological features are discussed along with a brief review of the literature. In addition, the importance of a differential diagnosis for this lesion is also discussed.

Effects of serial passage on the characteristics and chondrogenic differentiation of canine umbilical cord matrix derived mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are often known to have a therapeutic potential in the cell-mediated repair for fatal or incurable diseases. In this study, canine umbilical cord MSCs (cUC-MSCs) were isolated from umbilical cord matrix (n = 3) and subjected to proliferative culture for 5 consecutive passages. The cells at each passage were characterized for multipotent MSC properties such as proliferation kinetics, expression patterns of MSC surface markers and self-renewal associated markers, and chondrogenic differentiation. In results, the proliferation of the cells as determined by the cumulative population doubling level was observed at its peak on passage 3 and stopped after passage 5, whereas cell doubling time dramatically increased after passage 4. Expression of MSC surface markers (CD44, CD54, CD61, CD80, CD90 and Flk-1), molecule (HMGA2) and pluripotent markers (sox2, nanog) associated with self-renewal was negatively correlated with the number of passages. However, MSC surface marker (CD105) and pluripotent marker (Oct3/4) decreased with increasing the number of subpassage. cUC-MSCs at passage 1 to 5 underwent chondrogenesis under specific culture conditions, but percentage of chondrogenic differentiation decreased with increasing the number of subpassage. Collectively, the present study suggested that sequential subpassage could affect multipotent properties of cUC-MSCs and needs to be addressed before clinical applications.

Tunic morphology and viral surveillance in diseased Korean ascidians: Soft tunic syndrome in the edible ascidian, Halocynthia roretzi (Drasche), in aquaculture.

'Soft tunic syndrome' causes mass mortality in the edible ascidian Halocynthia roretzi in Korean and Japanese aquaculture. In histopathological comparison, there were no specific differences between diseased specimens from Korea and Japan, indicating that soft tunic syndrome occurring in Korea and Japan is the same disease. No bacterial or protozoan cells were microscopically detected in either healthy or diseased tunics suggesting they are not the direct causes of soft tunic syndrome. Attempts were made to isolate virus from affected ascidians taking into account temperature conditions in which soft tunic syndrome is most prevalent in the field. However, no viruses were isolated from diseased or non-diseased specimens using chinook salmon embryo (CHSE-214), flounder fin (FFN) or epithelioma papillosum cyprini (EPC) cell lines.

Antigenic differences of the scuticociliate Miamiensis avidus from Japan.

In Japan and Korea, outbreaks of scuticociliatosis have frequently occurred in Japanese flounder, Paralichthys olivaceus. Morphological observations and small subunit rRNA gene sequences have shown that the causative agent of scuticociliatosis in the flounder is Miamiensis avidus (syn. Philasterides dicentrarchi). In this study, we elucidated the antigenic differences between six Japanese M. avidus isolates as an initial step toward developing an effective vaccine against the disease. Four Japanese flounder isolates (IyoI, Nakajima, JF05To and Mie0301 isolates), one spotted knifejaw, Oplegnathus punctatus, isolate (SK05Kyo), and one ridged-eye flounder, Pleuronichthys cornutus, isolate (RF05To) were subjected to serological analysis. Antisera against IyoI, SK05Kyo, Nakajima and Mie0301 isolates were raised in rabbits and used for immobilization assays and Western blotting. Immobilization assays showed that the six isolates could be divided into three groups, tentatively designated serotype I for IyoI, JF05To, RF05To, SK05Kyo, serotype II for Nakajima and serotype III for Mie0301. Western blotting results supported these three serotypes, with marked similarities in the banding profiles of IyoI, JF05To, RF05To and SK05Kyo isolates, which were distinct from the Nakajima and Mie0301 isolates. Three isolates, IyoI, Nakajima and Mie0301 that were selected as representatives of each serotype, were highly pathogenic to Japanese flounder by experimental infection. Based on these findings, we propose that there are at least three M. avidus serotypes in Japan.

An impending crisis in the provision of histopathology expertise for mouse functional genomics.

The generation of new mouse models of human disease is accelerating rapidly, due to the completion of whole-genome sequencing efforts and technological advances in the manipulation of the mouse genome. We sought to investigate manpower issues in the provision of histopathology expertise for mouse functional genomics and compared this to the perceived demand from principal investigators (PIs). Through the European Commission (EC)-funded PRIME pathology training initiative, two questionnaires were devised to collect information from pathologists and EC-funded PIs on the current provision of mouse histopathology expertise in Europe and the demands for this service. We find that pathological analysis is being performed almost exclusively by professionally qualified pathologists, generally employed in clinical diagnostic posts, where the work is undertaken as collaboration outside of their contractual commitments but without previous training in veterinary or comparative pathology. The results indicate that there is a lack of both trainees and provision of specialist training in this field. Unsurprisingly, the availability of diagnostic expertise and advice falls far short of the number of genetically engineered mice (GEM) being generated for analysis. We analyse these results with reference to previous studies and discuss solutions for the future recruitment, training and funding for pathologists in mouse functional genomics in Europe.

Genetic characterization of the Korean porcine reproductive and respiratory syndrome viruses based on the nucleocapsid protein gene (ORF7) sequences.

To investigate the genetic characteristics of the Korean porcine reproductive and respiratory syndrome virus (PRRSV), we determined the complete sequence of the nucleocapsid protein gene (ORF7) from 105 PRRSV isolates from all nine Korean prefectures during the years 2003 through 2006. These sequences were then analyzed along with the published ORF7 sequences for two Korean PRRS viruses (PL97-1/1997 and LMY/2002) and 36 non-Korean viruses. The ORF7 nucleotide sequence identities among the 107 Korean PRRS viruses ranged from 86.2 to 100%, corresponding to 85.4 to 100% identity at the amino acid level. All of the Korean isolates examined belonged to the North American genotype. The ORF7 gene sequence from the North American prototype virus (VR-2332) and its derived vaccine virus (Ingelvac PRRS MLV) was 90.0-100% identical to the various ORF7 sequences of the Korean isolates, with corresponding amino acid identities from 91.0 to 100%. In the phylogenetic tree obtained by neighbor-joining analysis, all of the Korean PRRSVs were divided into four groups. Our ORF7 sequence data also revealed no correlations between the date or place of collection and the distribution of PRRSV in Korea. North American genotype PRRSVs may have been introduced into Korean swine herds some time ago; these viruses apparently radiated nationwide within a relatively short period of time. Within the North American genotype PRRSVs from around the world, the Korean PRRSVs did not emerge as a single independent clade overall, and their immediate relationships with the PRRSVs from other countries could not be determined.

Feed contaminated with classical swine fever vaccine virus (LOM strain) can induce antibodies to the virus in pigs.

In November 2004, antibodies to classical swine fever virus (csfv) were detected in finishing pigs during the annual serological surveillance in Jeju Province, Korea. In addition, csf vaccine viruses (lom strain) had recently been isolated from pigs raised on farms known to have csfv antibody-positive pigs. In contrast with mainland Korea, Jeju Province had been csf free and its pigs had not been vaccinated against csf for more than five years. An epidemiological investigation team from the National Veterinary Research and Quarantine Service investigated the current status of csf prevention on the Korean mainland and in Jeju Province to determine possible routes of introduction of the virus into the province. It was concluded that improperly processed blood meals, manufactured on mainland Korea, had been contaminated with the csf vaccine lom strain, and that the lom strain had been transmitted to pigs fed feed or feedstuffs containing the contaminated meal.

Medical physics aspects of cancer care in the Asia Pacific region.

Medical physics plays an essential role in modern medicine. This is particularly evident in cancer care where medical physicists are involved in radiotherapy treatment planning and quality assurance as well as in imaging and radiation protection. Due to the large variety of tasks and interests, medical physics is often subdivided into specialties such as radiology, nuclear medicine and radiation oncology medical physics. However, even within their specialty, the role of radiation oncology medical physicists (ROMPs) is diverse and varies between different societies. Therefore, a questionnaire was sent to leading medical physicists in most countries/areas in the Asia/Pacific region to determine the education, role and status of medical physicists.Answers were received from 17 countries/areas representing nearly 2800 radiation oncology medical physicists. There was general agreement that medical physicists should have both academic (typically at MSc level) and clinical (typically at least 2 years) training. ROMPs spent most of their time working in radiotherapy treatment planning (average 17 hours per week); however radiation protection and engineering tasks were also common. Typically, only physicists in large centres are involved in research and teaching. Most respondents thought that the workload of physicists was high, with more than 500 patients per year per physicist, less than one ROMP per two oncologists being the norm, and on average, one megavoltage treatment unit per medical physicist.There was also a clear indication of increased complexity of technology in the region with many countries/areas reporting to have installed helical tomotherapy, IMRT (Intensity Modulated Radiation Therapy), IGRT (Image Guided Radiation Therapy), Gamma-knife and Cyber-knife units. This and the continued workload from brachytherapy will require growing expertise and numbers in the medical physics workforce. Addressing these needs will be an important challenge for the future.

N-acetylphytosphingosine-induced apoptosis of Jurkat cells is mediated by the conformational change in Bak.

N-acetylphytosphingosine (NAPS), a sphingolipid derivative, is one of the well-known signal molecules that mediates various cellular functions, including cell growth, differentiation, and apoptosis. In this study, we demonstrated that NAPS induces apoptosis of Jurkat cells by activating Bak, but not Bax, which are both members of a proapoptotic subfamily of the Bcl-2 proteins. NAPS activated caspase-8 in a FADD-independent manner, but the lack of caspase-8 did not suppress the activation of caspase-3 and -9 and cell death, indicating that caspase-8 activation does not play an important role in NAPS-induced cell death. The overexpression of Bcl-xL, an anti-apoptotic protein, completely inhibited the activation of the caspases and apoptosis, assuming that NAPS-induced apoptosis was initiated by the mitochondria. The expression levels of pro- and anti-apoptotic Bcl-2 family members were not changed by the NAPS treatment. However, Bad was translocated from the cytosol into the mitochondria, where it bound to Bcl-xL, and Bak was dissociated from Bcl-xL and conformationally changed. Taken together, these findings indicate that NAPS induced apoptosis of Jurkat cells in a mitochondria-dependent manner that was controlled by the translocation of Bad and the conformational change in Bak.