bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mesenchymal stem cells of human placenta and umbilical cord suppress T‐cell proliferation at G0 phase of cell cycle
Authors: Vellasamy Shalini , Sandrasaigaran Pratheep , Vidyadaran Sharmili , Abdullah Maha , George Elizabeth , Ramasamy Rajesh 
Published: August 2013

AbstractMesenchymal stem cells (MSC) generated from human umbilical cord (UC‐MSC) and placenta (PLC‐MSC) were assessed and compared for their immunomodulatory function on T cells proliferation by analysis of the cell cycle. Mitogen stimulated or resting T cells were co‐cultured in the presence or absence of MSC. T‐cell proliferation was assessed by tritiated thymidine (3H‐TdR) assay and the mechanism of inhibition was examined bycell cycle and apoptosis assay. Both UC‐MSC and PLC‐MSC profoundly inhibited the proliferation of T‐cell, mainly via cell‐to‐cell contact. MSC‐mediated anti‐proliferation does not lead to apoptosis,but prevented T cells from entering S phase and they therefore accumulated in the G0/G1 phases. The anti‐proliferative activity of MSC was related to this cell cycle arrest of T‐cell. UC‐MSC produced a greater inhibition than PLC‐MSC in all measured parameters.
Preparation of Kupffer cell enriched non‐parenchymal liver cells with high yield and reduced damage of surface markers by a modified method for flow cytometry
Authors: Xu Fan , Zhen Peng , Zheng Yu , LIjuan Feng , Aiting Yang , Min Cong , Hong You , Jidong Jia 
Published: August 2013

AbstractThe aim of this study was to optimise a collagenase perfusion protocol for the isolation of a liver non‐parenchymal cell (NPC) suspension enriched for Kupffer cells that reduced damage to F4/80 antigen cell surface expression to allow analysis by flow cytometry.Kupffer cell‐enriched liver NPCs were isolated from C57BL/6 mice using different protocols. Flow cytometry was used to examine the effect of collagenase digestion on F4/80 expression on Kupffer cells, and results were represented by the percentage of F4/80 positive cells and by the F4/80 mean fluorescence intensity (MFI). The perfusion temperature, concentration of collagenase solution and total dosage of collagenase for liver perfusion influenced the effect of collagenase perfusion on the expression of F4/80 antigen on Kupffer cells. Collagenase perfusion at 28°C resulted in an increased percentage of F4/80 positive cells (P = 0.001) and MFI (P = 0.005) compared with 37°C. Perfusion with a total dose of 1.0 g/kg BW collagenase (using a 0.75 mg/mL solution) resulted in the highest percentage of F4/80 positive cells (P = 0.001) compared with 0.8 g/kg BW and 1.2 g/kg BW collagenase. Isolation of cells using the modified protocol resulted in a higher percentage of Kupffer cells (P < 0.001) and a higher MFI of F4/80 antigen (P < 0.001) compared with the common protocol.
Long‐term muscle‐derived cell culture: multipotency and susceptibility to cell death stimuli
Authors: Kalvelyte Audrone , Krestnikova Natalija , Stulpinas Aurimas , Bukelskiene Virginija , Bironaite Daiva , Baltriukiene Daiva , Imbrasaite Ausra 
Published: August 2013

AbstractImprovement in the yield of adult organism stem cells, and the ability to manage their differentiation and survival potential are the major goals in their application in regenerative medicine and in the adult stem cell research. We have demonstrated that adult rabbit muscle‐derived cell lines with an unlimited proliferative potential in vitro can differentiate into myogenic, osteogenic, adipogenic and neurogenic lineages. Studies of cell survival in vitro showed that differentiated cells, except neurogenic ones, are more resistant to apoptosis inducers compared to proliferating cells. Resistance to death signals correlated with the level of protein kinase AKT phosphorylation. Skeletal muscle‐derived cell lines can be multipurpose tools in therapy. Enhanced resistance of differentiated cells to certain types of damage shows their potential for long‐term survival and maintenance in an organism.This article was published online on 29 January 2013. An error was subsequently identified. This notice is included in the online and print versions to indicate that both have been corrected 6 March 2013.
Engraftment of genetically modified human amniotic fluid‐derived progenitor cells to produce coagulation factor IX after in utero transplantation in mice
Authors: Yang Chen‐Min , Gong Xiu‐Li , Qiu Jin , Tang Hui‐Xiang , Gong Zhi‐Juan , Huang Shu‐Zhen , Zeng Fanyi 
Published: August 2013

AbstractHuman amniotic fluid derived progenitor cells (hAFPCs) may be multipotent and can be considered a potential tool in the field of cell therapy for haemophilia B. Their capacity to express human coagulation factor IX (hFIX) after transduction and their fate after in utero transplantation is unknown. hAFPCs isolated from second trimester pregnancies were assessed for their phenotypic markers, multilineage capacity, and expression of hFIX after transduction. Their engraftment potential was analysed in a mouse model after in utero transplantation at embryonic day 12.5. Immunohistochemistry, fluorescence in situ, ELISA and PCR were used to assess post‐transplant chimeras. hAFPCs expressed several pluripotent markers, including NANOG, SOX2, SSEA4 and TRA‐1‐60, and could differentiate into adipocytes and osteocytes. In vitro, after transduction with hFIX and EGFP cDNAs, constitutive hFIX protein expression and clotting activity were found. Engraftment was achieved in various foetal tissues after in utero transplantation. Safe engraftment without oncogenesis was confirmed, with low donor cell levels, but persistent engraftment, into different organs (liver, heart and lung) through to 12 weeks of age. Transgenic expression of circulating hFIX was detected in recipient mice for up to 12 weeks. hAFPCs can be engrafted long‐term in immunocompetent mice after in utero transplantation. Thus, cell transplantation approaches using genetically engineered hAFPCs may prove valuable for the prenatal treatment for haemophilia B.
c‐Myc suppressed E‐cadherin through miR‐9 at the post‐transcriptional level
Authors: Liu Mei , Zhu Hongxia , Yang Shangbin , Wang Zaozao , Bai Jinfeng , Xu Ningzhi 
Published: August 2013

Abstractc‐Myc oncoprotein is overexpressed in most human cancers and regulates different genes and pathways in different cell types. E‐cadherin expression is repressed by MYC through a post‐transcriptional mechanism, but the exact mechanism remains elusive. Since E‐cadherin is a direct target of miR‐9 and miR‐9 can be activated by MYC and MYCN, this suggests that c‐Myc negatively modulates E‐cadherin through a microRNA pathway. We have established a c‐Myc‐inducible expression system in which the protein level and transcriptional activity of c‐Myc is significantly upregulated upon doxycycline induction. Overexpressed c‐Myc led to an EMT‐like conversion in the T‐REx‐293 cells and resulted in a significant decrease in E‐cadherin and an increase in Vimentin. Stem‐loop RT‐PCR showed elevated expression of miR‐9 when c‐Myc was induced to be overexpressed. Regarding the relationship of c‐Myc, miR‐9 and E‐cadherin, the expression of miR‐9 was curtailed by using antagomir‐9 in induced overexpressing c‐Myc. Restoration of E‐cadherin expression became much stronger in the presence of c‐Myc. Thus c‐Myc represses E‐cadherin at the post‐transcriptional level through miR‐9.
PPAR‐γ agonist pioglitazone prevents apoptosis of endothelial progenitor cells from rat bone marrow
Authors: Zhang Hui‐Feng , Wang Li , Yuan Hui‐Juan , Ma Yue‐Hua , Wang Yan‐Fang , Hu Zi‐Ying , Su Yong , Zhao Zhi‐Gang 
Published: August 2013

AbstractSelective peroxisome proliferator‐activated receptor‐γ (PPAR‐γ) agonist affects the functions of endothelial progenitor cells (EPCs). This study explores the effect of selective PPAR‐γ agonist, pioglitazone, on EPC apoptosis. The cells were cultured and identified via the double staining method in a medium containing different concentrations of pioglitazone. EPC apoptosis was detected by flow cytometry. On Day 7, EPCs engulfed DiL‐ac‐LDL and FITC‐UEA‐1, and showed yellow fluorescence in a laser‐scanning confocal microscope. EPC apoptosis inhibition was maximal at 50 µmol/L. The ability of pioglitazone to prevent EPC apoptosis may be mediated by the PI3K/Akt signal pathway. The use of thiazolidine two ketone (TZD) to reduce EPC apoptosis may have some potential in treating vascular diseases.
Curcumin attenuates cyclooxygenase‐2 expression via inhibition of the NF‐κB pathway in lipopolysaccharide‐stimulated human gingival fibroblasts
Authors: Hu Ping , Huang Ping , Chen Min Wei 
Published: August 2013

AbstractPorphyromonas gingivalis lipopolysaccharide (LPS) induces the expression of the cyclooxygenase‐2 (COX‐2), which contributes to the process of periodontitis. Curcumin, a constituent of turmeric, exhibits anti‐inflammatory properties. We have investigated the anti‐inflammatory effect of curcumin in human gingival fibroblasts (HGFs) stimulated by P. gingivalis LPS and its mechanism of action. HGFs pretreated with curcumin were stimulated by P. gingivalis LPS. COX‐2 mRNA and protein expressions were analysed by real‐time PCR and Western blot analysis. Activation of nuclear factor kappa B (NF‐κB) was analysed by the NF‐κB‐dependent luciferase activity and electrophoretic mobility‐shift assay (EMSA). Curcumin inhibited COX‐2 mRNA and protein synthesis in LPS‐stimulated HGFs in a dose‐dependent manner. P. gingivalis LPS activated NF‐κB‐dependent transcription in HGFs, which were also downregulated by pretreatment with curcumin. Therefore, curcumin can inhibit P. gingivalis LPS‐induced COX‐2 expression, which may be due to the inhibition of the NF‐κB pathway.
Functional compartmentalisation of NF‐κB‐associated proteins in A431 cells
Authors: Bolshakova Anastasia , Magnusson Karl‐Eric , Pinaev George , Petukhova Olga 
Published: August 2013

AbstractNF‐κB proteins belong to a family of ubiquitous transcription factors involved in a number of cellular responses. While the pathways of NF‐κB activation and input into the regulation of gene activity have been comprehensively investigated, its cytoplasmic functions are poorly understood. In this study we addressed effects of the compartmentalisation of NF‐κB proteins RelA/p65 and p50 in relation to the inhibitor IκB‐α, using fibronectin (FN) and epidermal growth factor (EGF) for environmental stimulation of epidermoid carcinoma A431 cells. We thus assessed the presence of NF‐κB family proteins in the cytosol, membrane, nuclear and cytoskeletal fractions with a special attention to the cytoskeletal fraction to define whether NF‐κB was active or not. Sub‐cellular fractionation demonstrated that the proportion of RelA/p65 differed in diverse sub‐cellular fractions, and that the cytoskeleton harboured about 7% thereof. Neither the nuclear nor the cytoskeleton fraction did contain IκB‐α. The cytoskeleton binding of RelA/p65 and p50 was further confirmed by co‐localisation and electron microscopy data. During 30‐min EGF stimulation similar dynamics were found for RelA/p65 and IκB‐α in the cytosol, RelA/p65 and p50 in the nucleus and p50 and IκB‐α in the membrane. Furthermore, EGF stimulation for 30 min resulted in a threefold accumulation of RelA/p65 in cytoskeletal fraction. Our results suggest that nuclear‐, membrane‐ and cytoskeleton‐associated NF‐κB are dynamic and comprise active pools, whereas the cytoplasmic is more constant and likely non‐active due to the presence of IκB‐α. Moreover, we discovered the existence of a dynamic, IκB‐α‐free pool of RelA/p65 associated with cytoskeletal fraction, what argues for a special regulatory role of the cytoskeleton in NF‐κB stimulation.
Role of altholactone in inducing type II apoptosis signalling pathway and expression of cancer‐related genes in cervical carcinoma HeLa cell line
Authors: Uthaisang‐Tanechpongtamb Wanlaya , Sriyabhaya Promjit , Wilairat Prapon 
Published: August 2013

AbstractGoniothalamus species (Annonaceae) is a shrub that grows in the rainforest of tropical Asia. Several compounds have been isolated and exhibit the potential use for cancer treatment. In this work, altholactone isolated from Goniothalamus macrophyllus was investigated for its cytotoxicity, apoptosis signalling and the expression of cancer‐related genes in the cervical carcinoma HeLa cells. Cytotoxicity was evaluated by MTT assay. Apoptotic characteristics were evaluated by morphological studies. Caspase‐3 activity was detected using a fluorogenic substrate. Cytochrome c release from mitochondria and protein Bid were determined by Western blotting and cancer‐related genes expression by RT‐PCR. The results demonstrated that altholactone was cytotoxic to HeLa (IC50 = 9.6 μg/mL), and apoptotic cell death was manifested by appearance of chromatin condensation and caspase‐3 activation, which was inhibited by specific inhibitors of both caspase‐8 and ‐9. Release into the cytosol of cytochrome and cleavage of Bid occurred. Altholactone also caused a decrease in bcl‐2 and an increase in p53 expression. These unique properties of altholactone suggest a potential for cancer chemotherapy.
Calpains are required for invasive and metastatic potentials of human HCC cells
Authors: Chen Bo , Tang Juan , Guo Yun‐Shan , Li Yong , Chen Zhi‐Nan , Jiang Jian‐Li 
Published: August 2013

AbstractCalpains are a conserved family of calcium‐dependent cysteine proteinases involved in various cellular functions. Two ubiquitous isoforms, µ‐ and m‐calpain, are key members of the calpain family that play essential roles in regulating cell migration and invasion. However, it remains unclear whether they are involved in the progression of hepatocellular carcinoma (HCC). Here, we investigated the functions of µ‐ and m‐calpain in the invasive and metastatic processes of human hepatoma cells. Our results indicated that the expression levels of calpains were elevated in HCC cells compared with those in normal hepatic cells. Our results indicated that small interfering RNA (siRNA)‐mediated silencing of µ‐ and m‐calpain expressions significantly suppressed the adhesive, migrative and invasive potentials of human hepatoma cells. The matrix metalloproteinases (MMPs) are key regulators of malignant tumour invasion and metastasis. siRNA‐mediated down‐regulation of µ‐ and m‐calpain expressions also significantly attenuated MMP‐2 and MMP‐9 secretion. Thus µ‐ and m‐calpain may play important roles in the invasion and metastasis of human hepatoma cells, and calpains may be drug targets for preventing HCC metastasis.
Identification of radiation‐sensitive expressed genes in the ICR and AKR/J mouse thymus
Authors: Bong Jin Jong , Kang Yu Mi , Shin Suk Chul , Choi Seung Jin , Lee Kyung Mi , Kim Hee Sun 
Published: August 2013

AbstractWe have investigated radiation‐sensitive expressed genes (EGs), their signal pathways, and the effects of ionizing radiation in the thymus of ICR and AKR/J mice. Whole‐body and relative thymus weights were taken and microarray analyses were done on the thymuses of high‐dose‐rate (HDR, 137Cs, 0.8 Gy/min, a single dose of 4.5 Gy) and low‐dose‐rate (LDR, 137Cs, 0.7 mGy/h, a cumulative dose of 1.7 Gy) irradiated ICR and AKR/J mice. Gene expression patterns were validated by quantitative polymerase chain reaction (qPCR). The effect of ionizing radiation on thymus cell apoptosis was measured terminal deoxynucleotidyl‐transferase‐mediated dUTP‐end labeling (TUNEL). LDR‐irradiation increased the mean whole‐body weight, but decreased the relative thymus weight of AKR/J mice. Radiation‐sensitive EGs were found by comparing HDR‐ and LDR‐irradiated ICR and AKR/J mice. qPCR analysis showed that 12 EGs had dose and dose‐rate dependent expression patterns. Gene‐network analysis indicated that Ighg, Igh‐VJ558, Defb6, Reg3g, and Saa2 may be involved in the immune response, leukocyte migration, and apoptosis. Our data suggest that expression of the HDR (Glut1, Glut4, and PKLR) and LDR radiation‐response genes (Ighg and Igh‐VJ558) can be dose or dose‐rate dependent. There was an increased number of apoptotic cells in HDR‐irradiated ICR mice and LDR‐irradiated AKR/J mice. Thus, changes of the mean whole‐body weight and relative thymus weight, EGs, signal pathways, and the effects of ionizing radiation on the thymus of ICR and AKR/J mice are described.
Effect of monochromatic visible light on intracellular superoxide anion production and mitochondrial membrane potential of B16F1 and B16F10 murine melanoma cells
Authors: Sato Kazuomi , Minai Yuji , Watanabe Hiroyuki 
Published: August 2013

AbstractWe have investigated the effect of visible light on animal cells using light‐emitting diodes to emit monochromatic visible light (red, yellow, green and blue light). To explore the relevant mechanism of apoptosis, we assessed the intracellular superoxide anion production and mitochondrial membrane potential (ΔΨm) of B16F1 and B16F10 murine melanoma cells after monochromatic light irradiation. Blue light caused ΔΨm depolarization subsequent to elevation of intracellular superoxide production. However, red and yellow light had no affect on both cell lines. Green light induced ΔΨm collapse only in B16F1 melanoma cells. ΔΨm is a key indicator of mitochondrial function, therefore its disruption causes mitochondria‐dependent apoptosis. Thus, blue light causes mitochondrial dysfunction and subsequent cell death.
Nitric oxide protects endothelium from cadmium mediated leakiness
Authors: Nagarajan Shunmugam , Rajendran Saranya , Saran Uttara , Priya M. Krishna , Swaminathan Akila , Siamwala Jamila H. , Sinha Swaraj , Veeriah Vimal , Sonar Punam , Jadhav Vivek , Jaffar Ali B.M. , Chatterjee Suvro 
Published: August 2013

AbstractCadmium targets the vascular endothelium causing endothelial dysfunction and leakiness of endothelial barrier. Nitric oxide plays a major role in mediating endothelial functions including angiogenesis, migration and permeability. The present study investigates the nitric oxide effects on cadmium induced endothelial leakiness. Results of ex vivo and in vitro permeability assays showed that even a sub‐lethal dose of cadmium chloride (1 µM) was sufficient to induce leakiness of endothelial cells. Cadmium drastically altered the actin polymerisation pattern and membrane tension of these cells compared to controls. Addition of nitric oxide donor Spermine NONOate (SP) significantly blunted cadmium‐mediated effects and recover endothelial cells integrity. Cadmium‐induced cytoskeletal rearrangements and membrane leakiness are associated with the low nitric oxide availability and high reactive oxygen species generation. In brief, we show the protective role of nitric oxide against cadmium‐mediated endothelial leakiness.
Regulation of expression of HGF in BM‐MSCs by baculovirus‐mediated transduction
Authors: Tu Yi Ji , Ye Ai Fang , Pan Zhi Min , Zheng Chao , Wu Tian Long , Cheng Xi Gao , Guo Fei 
Published: August 2013

AbstractBone marrow‐derived mesenchymal stem cells (BM‐MSCs) transplantation is widely adopted for the curing of osteonecrosis of femoral head (ONFH) in recent years. Furthermore, it is known that introducing hepatocyte growth factor (HGF) into BM‐MSCs will greatly improve the therapeutic effect of stem‐cell therapy owing to the great angiogenic and anti‐fibrotic capabilities of HGF. However, continuing overexpression of HGF in vivo may cause sarcomas, such as Kaposi's sarcoma. Aiming at enhancing the therapeutic effect and preventing the side effects of HGF‐modified stem‐cell transplantation on ONFH, we sought to construct a gene regulation system to control HGF expression in BM‐MSCs rigorously and accurately. We selected baculovirus as the gene vector and introduced pTet‐on advanced system into that. Finally, a virus vector vAcrtTA2s‐Ptight‐HGF was successfully built and delivered into BM‐MSCs to regulate the accurate expression of HGF. As shown in the results, different levels of HGF expression were verified by ELISA and Western blot with different induction doses of doxycycline (DOX). There was a dose–response relationship between them, and the optimum dose of DOX to induce HGF expression in BM‐MSCs in vitro was 1 µg/mL. We conclude that it is feasible to regulate HGF expression in BM‐MSCs by baculovirus‐mediated one‐off transduction.
Differential activation and inhibition of RhoA by fluid flow induced shear stress in chondrocytes
Authors: Wan Qiaoqiao , Kim Seung Joon , Yokota Hiroki , Na Sungsoo 
Published: August 2013

AbstractPhysical force environment is a major factor that influences cellular homeostasis and remodelling. It is not well understood, however, as a potential role of force intensities in the induction of cellular mechanotransduction. Using a fluorescence resonance energy transfer‐based approach, we asked whether activities of GTPase RhoA in chondrocytes are dependent on intensities of flow‐induced shear stress. We hypothesized that RhoA activities can be either elevated or reduced by selecting different levels of shear‐stress intensities. The result indicates that C28/I2 chondrocytes have increased RhoA activities in response to high shear stress (10 or 20 dyn/cm2), whereas a decrease in activity was seen with an intermediate shear stress of 5 dyn/cm2. No changes were seen under low shear stress (2 dyn/cm2). The observed two‐level switch of RhoA activities is closely linked to the shear‐stress‐induced alterations in actin cytoskeleton and traction forces. In the presence of constitutively active RhoA (RhoA‐V14), intermediate shear stress suppressed RhoA activities, while high shear stress failed to activate them. In chondrocytes, expression of various metalloproteinases is, in part, regulated by shear and normal stresses through a network of GTPases. Collectively, the data suggest that intensities of shear stress are critical in differential activation and inhibition of RhoA activities in chondrocytes.
Effects of frozen timing on the spindle density, the angle between the polar body and spindle, and embryo development of intracytoplasmic sperm injection in mouse mature oocytes
Authors: Song Wen‐yan , Xin Zhi‐min , Jin Hai‐xia , Peng Zhao‐feng , Chen Xue‐mei , Shi Sen‐lin , Dai Shan‐jun , Sun Ying‐pu 
Published: August 2013

AbstractBetter pregnancy outcomes can be obtained by human mature oocyte vitrification, but many problems remain to be resolved in human mature oocyte vitrification. Since mature oocyte development possesses its own maturity cycle, there should be the optimal timing for mature oocyte vitrification. The purpose of this study was to observe the effects of frozen timing on the spindle density, the angle between the polar body and spindle, and embryo development of intracytoplasmic sperm injection (ICSI) in vitrified mouse mature oocytes and explore its possible mechanism. Mouse oocytes were randomly divided into three groups according to different frozen timing including Groups A, B, and C in which oocytes were vitrified within 2 h after ovum pick‐up, and 3–4 and 5–6 h after ovum pick‐up, respectively. Spindle‐related parameters were measured, ICSI was performed. The spindle occurrence rate of vitrified‐thawed oocytes was 98.4% in Group A, 82.3% in Group B, and 75.8% in Group C, without statistical differences between pre‐vitrification and post‐thawing and among the three groups (P > 0.05). The angles between the polar body and spindle were larger after thawing than before vitrification (P < 0.01). The spindle retardance values were lower after thawing than before vitrification in Groups B and C (P < 0.05), but higher in Group A (P < 0.05). The spindle retardance values before vitrification were higher in Group B than in Groups A and C (P < 0.05), but the spindle retardance value, oocyte survival and two‐cell rate after thawing were higher in Group A than in Groups B and C (P < 0.05). There were no statistical differences in ICSI fertility rate between the three groups (P > 0.05). The damage on the spindle is the slightest and embryo quality is the highest in the mouse oocytes vitrified within 2 h after ovum pick‐up. The spindle retardance value is more valuable than the spindle occurrence rate in the evaluation of vitrified‐thawed oocyte quality, and is positively correlated with embryo quality.