Liver cancer stem cells are selectively enriched by low-dose cisplatin

Accumulating evidence has indicated the importance of cancer stem cells in carcinogenesis. The goal of the present study was to determine the effect of low-dose cisplatin on enriched liver cancer stem cells (LCSCs). Human hepatoblastoma HepG2 cells were treated with concentrations of cisplatin ranging from 1 to 5 μg/mL. Cell survival and proliferation were evaluated using a tetrazolium dye (MTT) assay. LCSCs were identified using specific markers, namely aldehyde dehydrogenase-1 (ALDH1) and CD133. The percentage of ALDH1+ or CD133+ cells was examined by flow cytometric analysis. The expression of ALDH1 and/or CD133 in HepG2 cells was determined by immunocytochemical analysis. Low-dose cisplatin treatment significantly decreased cell survival in HepG2 cells after 24 or 72 h. However, the percentage of LCSCs in the surviving cells was greatly increased. The percentage of ALDH1+ or CD133+ cells was increased in a time- and dose-dependent manner after treatment with 1-4 μg/mL cisplatin, whereas 5 μg/mL cisplatin exposure slightly reduced the number of positive cells. These findings indicate that low-dose cisplatin treatment may efficiently enrich the LCSC population in HepG2 cells.

Therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia using Fe2O3 nanoparticles

This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3 nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3 MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.

Esculetin, a coumarin derivative, exerts in vitro and in vivo antiproliferative activity against hepatocellular carcinoma by initiating a mitochondrial-dependent apoptosis pathway

This study investigated the in vitro and in vivo antiproliferative activity of esculetin against hepatocellular carcinoma, and clarified its potential molecular mechanisms. Cell viability was determined by the MTT (tetrazolium) colorimetric assay. In vivoantitumor activity of esculetin was evaluated in a hepatocellular carcinoma mouse model. Seventy-five C57BL/6J mice were implanted with Hepa1-6 cells and randomized into five groups (n=15 each) given daily intraperitoneal injections of vehicle (physiological saline), esculetin (200, 400, or 700 mg·kg-1·day-1), or 5-Fu (200 mg·kg-1·day-1) for 15 days. Esculetin significantly decreased tumor growth in mice bearing Hepa1-6 cells. Tumor weight was decreased by 20.33, 40.37, and 55.42% with increasing doses of esculetin. Esculetin significantly inhibited proliferation of HCC cells in a concentration- and time-dependent manner and with an IC50 value of 2.24 mM. It blocked the cell cycle at S phase and induced apoptosis in SMMC-7721 cells with significant elevation of caspase-3 and caspase-9 activity, but did not affect caspase-8 activity. Moreover, esculetin treatment resulted in the collapse of mitochondrial membrane potential in vitro and in vivo accompanied by increased Bax expression and decreased Bcl-2 expression at both transcriptional and translational levels. Thus, esculetin exerted in vitro and in vivo antiproliferative activity in hepatocellular carcinoma, and its mechanisms involved initiation of a mitochondrial-mediated, caspase-dependent apoptosis pathway.

Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication

Bone homeostasis seems to be controlled by delicate and subtle “cross talk” between the nervous system and “osteo-neuromediators” that control bone remodeling. The purpose of this study was to evaluate the effect of interactions between neuropeptides and human bone morphogenetic protein 2 (hBMP2) on human osteoblasts. We also investigated the effects of neuropeptides and hBMP2 on gap junction intercellular communication (GJIC). Osteoblasts were treated with neuropeptide Y (NPY), substance P (SP), or hBMP2 at three concentrations. At various intervals after treatment, cell viability was measured by the MTT assay. In addition, cellular alkaline phosphatase (ALP) activity and osteocalcin were determined by colorimetric assay and radioimmunoassay, respectively. The effects of NPY, SP and hBMP on GJIC were determined by laser scanning confocal microscopy. The viability of cells treated with neuropeptides and hBMP2 increased significantly in a time-dependent manner, but was inversely associated with the concentration of the treatments. ALP activity and osteocalcin were both reduced in osteoblasts exposed to the combination of neuropeptides and hBMP2. The GJIC of osteoblasts was significantly increased by the neuropeptides and hBMP2. These results suggest that osteoblast activity is increased by neuropeptides and hBMP2 through increased GJIC. Identification of the GJIC-mediated signal transduction capable of modulating the cellular activities of bone cells represents a novel approach to studying the biology of skeletal innervation.

Standing before a sentence : Moore’s paradox and a perspective from within language

Med avstamp i ett satsbegrepp som inspirerats av Ludwig Wittgenstein, där sats och kontext betraktas som internt förbundna och där den verkliga användningen av en sats är central för dess mening, visar avhandlingen hur filosofers olika uppfattning om den filosofiska terminologins roll och möjligheter har följder för deras sätt att uppfatta och behandla filosofiska frågeställningar. Moores paradox fungerar som testfall. Denna kända filosofiska frågeställning har diskuterats sedan 1940-talet och kretsar kring hävdandet av satsen ”Jag tror att det regnar och det regnar inte”. Problemet är att det vore märkligt för en talare att hävda satsen om sig själv: det verkar finnas ett logiskt hinder för att hävda en sats även om den är välformad, kunde vara sann och inte på ett entydigt sätt innehåller en kontradiktion. Moores paradox behandlas genom att granska några lösningsförslag (framförda av bl.a. G.E. Moore, J. L. Austin, J. Searle) och de uppfattningar om språket som de förutsätter. Genom att kontrastera dessa med ett användarperspektiv där språkets användning i meningsfulla sammanhang förväntas spela en central roll i behandlingen av paradoxen visar författaren hur också den intellektuella kontext inom vilken en filosof betraktar paradoxen spelar en avgörande roll för vilka mått som krävs för att upplösa den. Istället för att föreslå en egen lösning presenteras en behandling av paradoxen genom en diskussion och utredning av de grundvillkor som leder till att den uppstår. Här intar satsbegreppet en central plats. Avhandlingen är författad inom traditionen efter den senare Wittgenstein (närmare bestämt den gren som ansluter sig till en s.k. terapeutisk filosofisyn) men går i dialog med filosofiskt arbete som ligger utanför traditionen. Författaren strävar både till att medla mellan, sammanföra och särskilja olika temata som behandlats på ett sätt inom traditionen och på andra sätt utanför den och använder sig här av metaforen att filosofera inifrån i kontrast till utanför språket. Avhandlingens huvudsakliga bidrag är till metafilosofin genom dess fokus på filosofins metod. ------------------------------------------------------------ Mooren paradoksi on tunnettu filosofinen ongelma, josta on keskusteltu 1940-luvulta alkaen. Paradoksin keskeisenä osana esiintyy väitelause ”Uskon, että ulkona sataa, mutta ulkona ei sada”. Ongelman lähtökohta on, että olisi merkillistä jos puhuja esittäisi lauseen itseään koskevana väitteenä: väite näyttää olevan loogisesti mahdoton esittää, vaikka se on hyvinmuodostettu, mahdollisesti tosi, eikä yksiselitteisellä tavalla sisällä kontradiktiota. Tässä väitöskirjassa Mooren paradoksi esiintyy esimerkkinä filosofisesta ongelmasta, jossa mm. lausekäsitteellä on keskeinen rooli ja jonka avulla voidaan muodostella ns. kielensisäinen filosofinen näkökulma. Mooren paradoksia käsitellään tarkastelemalla muutamia ratkaisuehdotuksia (mm. G.E. Mooren, J.L. Austinin, J. Searlen esittämät) ja niitä kielikäsityksiä joita nämä ratkaisuehdotukset edellyttävät. Asettamalla vastakkain ratkaisuehdotukset ja käyttäjänäkymä, jossa kielenkäyttö mielekkäissä konteksteissa asetetaan keskeiseen asemaan paradoksin käsittelyssä, kirjoittaja osoittaa miten myös paradoksin filosofisen tarkastelun intellektuaalinen konteksti on hyväksyttävien toimenpiteiden löytämisen kannalta ratkaisevaa. Uuden ratkaisun sijaan kirjoittaja ehdottaa paradoksin terapeuttista käsittelyä: keskustelua ja selontekoa niistä perusehdoista, jotka johtavat sen syntyyn. Tässä käsittelyssä lausekäsite on keskipisteenä – erityisesti Ludwig Wittgensteinin inspiroima lausekäsitys, jossa lauseen mielekkyyden kannalta sen todellinen käyttö mielekkäässä kontekstissa on ratkaiseva tekijä. Väitöskirja liittyy myöhäisiwittgensteinilaiseen perinteeseen, johon kuuluu ns. terapeuttinen filosofiankäsitys, mutta teos keskustelee myös tradition ulkopuolisten ajattelijoiden kanssa yrittäen yhdistää, sovitella ja myös eritellä teemoja, joita käsitellään yhdellä tavalla tradition sisällä ja toisella tavalla sen ulkopuolella. Väitöskirjan pääasiallinen panos sijoittuu metafilosofiaan, koska sen mielenkiinnon kohteena ovat erityisesti filosofian menetelmät.

Customer Experience Management – How to Improve the Customer Experience Management and its Measurement

The main goal of this master’s thesis was to find out, how to improve customer experience management and measurement. This study is a qualitative case study, in which the data collection method has been interviews. In addition, some of the company’s customer experience measurement methods have been analyzed. The theoretical background is applied in practice by interviewing 5 representatives from the case company. In the case company, the management has launched a customer experience focused program, and given guidelines for customer experience improvement. In the case company, customer experience is measured with different methods, one example is asking the recommendation readiness from a customer. In order to improve the customer experience management, the case company should define, what the company means with customer experience and what kind of customer experience the company is aiming to create. After the encounter, the customer should be left with feelings of satisfaction, positivity and trust. The company should focus on easiness in its processes, on top of which the processes should work fluently. The customer experience management should be improved through systematic planning, and by combining and standardizing different measures. In addition, some channel-based measures should be used. The measurement conducted should be more customer focused, and the case company should form an understanding, which touch points are the most relevant to measure.