bluuebunny/crossref_metadata_2025_split huggingface dataset:
Integrin alpha v beta 6 contributes to maintaining corneal epithelial barrier function
Authors: Guo Huiling , Du Gaiping , Wang Liqiang , Wang Dajiang , Hu Lianna , Huang Yifei 
Published: August 2013

AbstractThe corneal epithelial barrier dysfunction is associated with a number of disorders of the cornea. This study aims to investigate the role of integrin alpha v beta 6 (avb6) in maintaining the corneal epithelial barrier function. In this study, the association of avb6 and endosome/lysosome in the human corneal epithelial cell line, HCE cells, was observed with immunocytochemistry. The corneal epithelial barrier function was assessed with HCE monolayers in Transwells. The results showed that avb6 was observed attaching to endosomes in HCE cells. Knockdown of avb6 resulted in disturbing the fusion of endosome/lysosome. Tumor necrosis factor (TNF)‐α could bind avb6 to form a complex that interfered with the fusion of endosome/lysosome in HCE cells. With the HCE monolayer as a corneal epithelial barrier model, the knockdown of avb6 or exposure to TNF‐α markedly increased the epithelial barrier permeability to a macomolecular protein, ovalbumin, in Transwell system. After passing through the deficient epithelial barrier, the ovalbumin molecules still conserved the antigenicity. We conclude that the integrin avb6 plays an important role in the fusion of endosome/lysosome in corneal epithelial cells; inhibition of avb6 results in corneal epithelial barrier dysfunction.
Effect of substrate stiffness on the osteogenic differentiation of bone marrow stem cells and bone‐derived cells
Authors: Witkowska‐Zimny Malgorzata , Walenko Katarzyna , Wrobel Edyta , Mrowka Piotr , Mikulska Agnieszka , Przybylski Jacek 
Published: August 2013

AbstractThere is a profound dependence of cell behaviour on the stiffness of its microenvironment. To gain a better understanding of the regulation of cellular differentiation by mechanical cues, we investigated the influence of matrix stiffness (E = 1.46 kPa and E = 26.12 kPa) on differentiated osteogenic cell lineage of bone marrow stem cells (BM‐MSCs) and bone‐derived cells (BDCs) using flexible collagen‐coated polyacrylamide substrates. Differentiation potential was determined by measuring alkaline phosphatase activity, expression of osteoblast‐specific markers including alkaline phosphatase, osteocalcin, Runx2 and collagen type I, as well as assessment of mineralisation (Alizarin Red S staining). We found that osteogenic differentiation can be regulated by the rigidity of the substrate, which may depend on the commitment in multi‐ or uni‐potent targeting cells. Osteogenic differentiation of BM‐MSCs was enhanced on a stiff substrate compared to a soft one, whereas BDCs osteogenic differentiation did not vary depending on the substrate stiffness. The data help in understanding the role of the external mechanical determinants in stem cell differentiation, and can also be useful in translational approach in functional tissue engineering.
Functional impact of cholesterol sequestration on actin cytoskeleton in normal and transformed fibroblasts
Authors: Chubinskiy‐Nadezhdin Vladislav I. , Efremova Tatiana N. , Khaitlina Sofia Y. , Morachevskaya Elena A. 
Published: August 2013

AbstractMembrane cholesterol and lipid rafts are implicated in various signalling processes involving actin rearrangement in living cells. However, functional link between raft integrity and organisation of cytoskeleton remains unclear. We have compared the effect of cholesterol sequestration on F‐actin structures in normal and transformed fibroblasts in which microfilament system is developed to a different extent. The depletion of membrane cholesterol by methyl‐beta‐cyclodextrin (MbCD) resulted in a disruption of lipid rafts in plasma membrane as it was revealed by fluorescent labelling of GM1 ganglioside. In normal fibroblasts with highly developed microfilament system, the cholesterol depletion resulted in actin disassembly and reduction of stress fibres. However, in transformed cells containing low amount of fibrillar actin, MbCD treatment induced intensive formation of stress fibres and increased cell spreading. The results show that the effect of cholesterol depletion and lipid raft disruption on microfilament system is critically determined by the initial state of cytoskeleton, specifically, by the balance of polymerised and monomeric actin in the cell. We assume that uncapping of the microfilaments is the key step of cholesterol‐regulated actin remodelling.
Computational prediction and characterisation of ubiquitously expressed new splice variant of Prkaca gene in mouse
Authors: Banday Abdul Rouf , Azim Shafquat , Hussain Mohammed Aamir , Nehar Shamshun , Tabish Mohammad 
Published: August 2013

AbstractPrkaca gene of mouse encodes for a cAMP dependent protein kinase catalytic alpha subunit. PKA occurs naturally as a 4‐membered structure having two regulatory (R) and two catalytic (C) subunits each encoded by separate gene. Alternatively spliced two transcript variants are known for the Prkaca gene, which encode for two isoforms of PKA C‐subunits, namely Cα1 and Cα2. These isoforms arise as a result of alternative splicing of the first coding exon with the internal exons. We have identified a new transcript variant using combinatorial approach of bioinformatics and molecular biology techniques involving RT‐PCR, semi‐nested PCR and sequencing. The new transcript variant encoding Cα3 isoform has N‐terminus that differs from Cα1 and Cα2 isoforms. Cα3 isoform also arise as a result of alternative splicing of first coding exon with the internal exon. Newly identified transcript is expressed ubiquitously in different tissues examined.
Evidence that TMEM67 causes polycystic kidney disease through activation of JNK/ERK‐dependent pathways
Authors: Du E , Li Hong , Jin Shunying , Hu Xuemei , Qiu Mengsheng , Han Ruifa 
Published: August 2013

AbstractTMEM67 mutations are associated with severe autosomal recessive polycystic kidney disease (ARPKD) in both humans and animals. However, the molecular mechanisms underlying the pathogenesis of PKD caused by TMEM67 mutations remain to be determined. We have investigated the possible signalling pathways involved in the pathogenesis of PKD. Overexpression of TMEM67 in human embryonic kidney (HEK293) cells triggered the activation of overall tyrosine phosphorylated proteins, extracellular signal‐regulated kinase (ERK) and c‐jun N‐terminal KINASE (JNK). Activation was suppressed by pharmacological inhibitors of ERK or JNK. Activation of the mammalian target of rapamycin (mTOR) or p70s kinase (S6K) did not occur, although elevated phosphorylation of eIF4E‐binding protein 1 (4E‐BP1), a target of S6K, was seen. In animal studies, activation of a variety of signalling molecules was linked to ERK, JNK and 4E‐BP1. Significant induction of phosphorylation of tyrosine phosphorylated proteins, ERK and 4E‐BP1, at different postnatal ages was detected in mutant kidneys of B6C3Fe a/a‐bpck mice, a cystic renal disease mouse model caused by TMEM67 loss of function mutation. Based on these in vitro and in vivo observations, we propose that TMEM67 mutations cause PKD through ERK‐ and JNK‐dependent signalling pathways, which may provide novel insight into the therapy of polycystic kidney diseases.
Intratracheal transplantation of bone marrow‐derived mesenchymal stem cells reduced airway inflammation and up‐regulated CD4+CD25+ regulatory T cells in asthmatic mouse
Authors: Ge Xiahui , Bai Chong , Yang Jianming , Lou Guoliang , Li Qiang , Chen Ruohua 
Published: August 2013

AbstractMesenchymal stem cells attenuate the severity of lung injury due to their immunomodulatory properties. The effect of bone marrow‐derived mesenchymal stem cells on asthma is seldom reported. We have examined the effect of BMSCs on airway inflammation in asthma. Forty female BALB/c mice were equally randomised into PBS group, BMSCs treatment group, BMSCs control group and asthmatic group. Reactivity of the airway to acetylcholine was measured by barometric plethysmography. Cytokine profiles of bronchoalveolar lavage fluid and serum were determined by enzyme‐linked immunosorbent assay. Morphometric analysis was done with haematoxylin and periodic‐acid Schiff staining. Engraftment of BMSCs in asthmatic mice significantly decreased the number of eosinophils and mononuclear cells in bronchoalveolar lavage fluid and the airway (P < 0.05). Both goblet cell hyperplasia and responsiveness to acetylcholine were significantly reduced in BMSCs treatment groups. Moreover, BMSCs engraftment caused significant increases the ratio of Treg in pulmonary lymph node and interleukin‐10 (IL‐10) and interleukin‐12 levels in BALF and serum. We conclude that BMSCs engraftment ameliorated airway inflammation and improved lung function in asthmatic mouse and the protective effect might be mediated by upregulating Treg and partly involved with increasing IL‐10.
Cold exposure lowers energy expenditure at the cellular level
Authors: Park Seyeon , Chun Sohyun , Kim Danuh 
Published: August 2013

AbstractMitochondrial function is intimately involved in various metabolic processes and is therefore essential to maintain cell viability. Of particular importance is the fact that mitochondrial membrane potential (ΔΨm) is coupled with oxidative phosphorylation to drive adenosine triphosphate (ATP) synthesis. We have examined the effects of cold temperature stress on ΔΨm and the role of cold temperature receptor expression on ΔΨm. Human bronchial endothelial cell line, BEAS‐2B, and human embryonic kidney, HEK293, cell line were transfected with the gene for cold temperature responsive receptor protein TRPM8 or TRPA1, and exposed to cold temperature. ΔΨm was monitored using 5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetraethylbenzimidazoyl carbocyanine iodide derivative (JC‐10), a ΔΨm probe. While cold temperatures significantly increased ΔΨm and mitochondrial ATP levels in cells transfected with temperature responsive receptor TRPM8 or TRPA1, no change was noted in wild‐type cells. Moreover, the change in ΔΨm and ATP level was a dynamic process. ΔΨm was raised to peak levels within10 min of cold exposure, followed by a return to baseline levels at 30 min. Our findings suggest that cold temperature exposure increased mitochondrial ΔΨm via a mechanism involving cold temperature receptors.
Altered VEGF‐A and receptor mRNA expression profiles, and identification of VEGF144 in foetal rat calvaria cells, in coculture with microvascular endothelial cells
Authors: Weyand Birgit , von Schroeder Herbert P. 
Published: August 2013

AbstractCellular proliferation and differentiation during angiogenesis and osteogenesis require the communication of different cell types through growth factors and their receptors. Vascular endothelial growth factor (VEGF‐A) plays an important role in osteoblast and endothelial cell intercommunication. We have investigated the effect of monocultures and indirect coculture of foetal rat calvarial (FRC) osteoblasts and microvascular endothelial cells (ECs) on nodule formation, proliferation, and mRNA‐expression of VEGF‐A and its receptors during culturing. Despite increased nodule formation in the presence of dexamethasone (Dex) in monocultures, the number of nodules and alkaline phosphatise activity were decreased in cocultured FRCs. VEGF mRNA expression over the differentiation period showed the expression of most Vegf isoforms is biphasic in both FRCs and ECs, whereas receptor expression was quite variable; however, that of Np‐2 in FRCs increased steadily and significantly from 8 h to 14 days after an initial drop in expression. Significant changes in the proportion of Vegfa by Day 14 were noted mainly in the matrix‐bound variants Vegf144 and Vegf188 in ECs and osteoblasts, respectively. Less striking results were seen in the expression of the soluble isoforms in either cell type. These results have identified expression of Vegf144 in osteoblasts, suggesting a possible autocrine and/or paracrine role that is affecting osteoblast mineralisation along with Vegf188, as well as possible early roles of these isoforms in initial cell attachment. Further study of VEGF expression in coculture and Vegf144 will lead to better understanding of its role in cell–cell communication and bone development.
NF‐κBP65 promotes invasion and metastasis of oesophageal squamous cell cancer by regulating matrix metalloproteinase‐9 and epithelial‐to‐mesenchymal transition
Authors: Wang Feng , He Wei , Fanghui Peng , Wang Liuxing , Fan Qingxia 
Published: August 2013

AbstractNF‐κB has been recognized as one of the factors responsible for the development of cancer; however, the mechanism by which high expression of NF‐κB contributes to the progression of human oesophageal squamous cell cancer (ESCC) is not fully understood. In our investigations, NF‐κBP65 was overexpressed in human ESCC tissues, especially in ESCC tissues with deep invasion and lymph node metastasis. Suppression of NF‐κBP65 by siRNA decreased the invasion and proliferation ability of EC9706 cells in vitro. Furthermore, siRNA‐mediated NF‐κBP65 knock‐down could lead to the downregulation of MMP‐9, a metastasis‐related gene. Reduced E‐cadherin is a hallmark of invasive carcinomas that have acquired epithelial–mesenchymal transition (EMT) phenotypes and Vimentin is another molecule that is used widely as a marker of the EMT. We found upregulation of E‐cadherin expression and downregulation of Vimentin was induced by NF‐κBP65 siRNA, which suggests that NF‐κBP65 siRNA could inhibit the invasion and proliferation ability of ECSS through attenuating the expression of MMP‐9 and EMT. Thus, ESCC NF‐κBP65 could be a useful target for cancer prevention and therapy.
Characteristics of Notch2+ pancreatic cancer stem‐like cells and the relationship with centroacinar cells
Authors: Zhou Zhu‐Chao , Dong Qiang‐Gang , Fu De‐liang , Gong Yi‐yi , Ni Quan‐Xing 
Published: August 2013

AbstractNotch2, a surface marker in cell lines, is used to isolate, identify and localise pancreatic cancer stem‐like cells and is a target for therapy of these cells. Sphere formation was induced in Panc‐1 and Bxpc‐3 pancreatic cancer cell lines, and Notch2+ cells were separated from Bxpc‐3 and Panc‐1 cell lines by magnetic activated cell sorting (MACS). Expression of stem cell‐related markers, OCT4, Nanog and PDX1, were measured by immunofluorescent (IF) staining. Expression of Notch2 was also determined immunohistochemically in pancreatic tissues. Notch2+ cells were transplanted in subcutaneous of mice. AQP1 and AQP5 were also measured by IF in Bxpc‐3 cells. The Notch signal pathway inhibitor, Compound E (CE), was used to treat Notch2+Bxpc‐3 cells, and their vitalities were subsequently measured by the CCK‐8 method. Positive expression of OCT4, Nanog and PDX1 was observed in Notch2+ cells. Notch2+ cells at centroacinar cell (CAC) and terminal ductal locations expressed AQP1 and AQP5. They were strongly tumourigenic in mice, and CE inhibited proliferation of Notch2+ Bxpc‐3 cells to some degree. OCT4 and Nanog can be used as markers of self‐renewal in pancreatic cancer stem cells. Notch2+ cells in human pancreatic cancer Bxpc‐3 and Panc‐1 cell lines had the properties of cancer stem cells. The results suggest that Notch2+ pancreatic cancer stem‐like cells had a close relationship with CAC.
RNA interference targeting CD147 inhibits the invasion of human cervical squamous carcinoma cells by downregulating MMP‐9
Authors: Fan Xiaobin , Wu Weiguang , Shi Haixia , Han Jianqiu 
Published: August 2013

AbstractCervical squamous carcinoma is a highly invasive tumour that has a great capacity to metastasise. Extracellular matrix metalloproteinase inducer (EMMPRIN or CD147), a member of the immunoglobulin superfamily, is a widely distributed cell surface glycoprotein. It is highly expressed on malignant tumour cell surfaces, including human cervical squamous carcinoma. It also plays a critical role in the invasive and metastatic activity of malignant cells by stimulating the expression of matrix metalloproteinases (MMPs). The anti‐invasive effect of small interfering RNA (siRNA) against CD147 on human cervical squamous carcinoma cells and its possible pathways has been investigated. The downregulation of CD147 by transfection with siRNA resulted in MMP‐9 expression and decreased activity in the cervical squamous carcinoma cell line SiHa. In vitro analysis showed that the invasive capacity of SiHa cells decreased. Thus CD147 inhibition and subsequent MMP‐9 deletion may have anti‐tumour effects by inhibiting the invasiveness of human cervical squamous carcinoma cells.
RNAi‐mediated MMP‐9 silencing inhibits mouse melanoma cell invasion and migration in vitro and in vivo
Authors: Tang Zhao‐Yong , Liu Yang , Liu Long‐Xing , Ding Xiao‐Yan , Zhang Hong , Fang Liao‐Qiong 
Published: August 2013

AbstractMMP‐9 participates in tumour growth, invasion, metastasis and vascularisation. Thus, inhibition of MMP‐9 may be involved in the process of tumourigenesis. We have investigated the effect of RNAi‐mediated MMP‐9 silencing on inhibiting invasion and migration of mouse melanoma cell B16. A specific and optimised siRNA vector was used to target MMP‐9 mRNA synthesis in B16 cells. Changes of invasion and migration capability of B16 cell were examined after transfection at different time, and a footpad tumour model was performed to measure the effect of MMP‐9 siRNA on melanoma tumourigenesis in vivo. In vitro, down‐regulation of MMP‐9 expression significantly inhibited B16 cell invasion and migration. In vivo, intratumoural injection of plasmid DNA expressing MMP‐9 siRNA every 3 days for three times remarkably inhibited melanoma growth and also suppressed tumour metastasis. The results indicate that RNA‐mediated targeting of MMP‐9 may have promising applications for the treatment of melanoma.
New strategies in cord blood cells transplantation
Authors: Yu Xin , Gu Zongjiang , Wang Yunliang , Wang Hongwei 
Published: August 2013

AbstractFor patients lacking a human leucocyte antigen‐matched donor, umbilical cord blood (UCB) is an ideal, alternative source of haematopoietic stem cells (HSCs) for transplantation purposes. UCB has many advantages over bone marrow or peripheral blood taken from volunteer donors. UCB is also an important source of other stem cells, including endothelial progenitors, mesenchymal stem cells, very small embryonic/epiblast‐like (VSEL) stem cells, and unrestricted somatic stem cells, which are potentially suitable for regenerative medicine. However, a significant clinical problem is that the number of HSCs in one cord‐blood unit is not enough for an adult transplantation. The development of new approaches including use of multiple donors, ex vivo expansion, increasing efficiency of homing and engraftment, retrieving more cells from the placenta and cord blood is of crucial importance for the delayed engraftment after UCB transplantation. In the future, UCB will emerge as a source of cells for cellular therapies associated with tissue repair and regeneration.
3D Arrays for high throughput assay of cell migration and electrotaxis
Authors: Zhao Sanjun , Gao Runchi , Devreotes Peter N. , Mogilner Alex , Zhao Min 
Published: August 2013

AbstractCell behaviour in 3D environments can be significantly different from those in 2D cultures. With many different 3D matrices being developed and many experimental modalities used to modulate cell behaviour in 3D, it is necessary to develop high throughput techniques to study behaviour in 3D. We report on a 3D array on slide and have adapted this to our electrotaxis chamber, thereby offering a novel approach to quantify cellular responses to electric fields (EFs) in 3D conditions, in different matrices, with different strains of cells, under various field strengths. These developments used Dictyostelium cells to illustrate possible applications and limitations.
Correlation analysis between gene expression profile of high‐fat emulsion‐induced non‐alcoholic fatty liver and liver regeneration in rat
Authors: Wang Gaiping , Li Bing , Hao Yunpeng , Zhi Jia , He Chunxiao , Xu Cunshuan 
Published: August 2013

AbstractTo explore the relevance of non‐alcoholic fatty liver disease (NAFLD) to liver regeneration (LR), rat models of non‐alcoholic steatohepatitis (NASH) and LR were established, respectively, then Rat Genome 230 2.0 Array was used to detect the gene expression abundance of them, and the reliabilities of the array data were confirmed by real‐time RT‐PCR. As a result, the expression of 93 genes was significantly changed during NAFLD occurrence and 948 genes in LR. Hierarchical clustering indicated that the expression profiles of the above two events were quite different. K‐means cluster classified their expression patterns into four clusters, and gene expression trends of clusters 1, 2 were similar in NAFLD and LR, while clusters 3, 4 were contrary with the gene expression changes of LR more abundant. DAVID classifications and functional enrichment analysis found that lipid metabolism and carbohydrate metabolism were stronger in NAFLD than in LR, but some other physiological activities including inflammation/immune response, cell adhesion, and migration, cell proliferation and differentiation in NAFLD were weaker than in LR. IPA further indicated that lipid metabolism, inflammation response, and cellular development were highly associated with NAFLD, and thus identified some potential biomarkers for NAFLD.
Sphere‐forming cells from peripheral cornea demonstrate polarity and directed cell migration
Authors: Yoon Jinny Jung , Wang Ellen F. , Ismail Salim , McGhee Jennifer J. , Sherwin Trevor 
Published: August 2013

AbstractSphere‐forming cells from peripheral cornea represent a potential source of progenitor cells for treatment of corneal degenerative diseases. Control of cellular repopulation on transplantable substrates is important to prevent uncontrolled growth in unfavourable directions. The coordination of cellular outgrowth may be in response to environmental cues and/or cellular signals from other spheres. To investigate this, cell migration patterns were observed following placement of spheres on an adhesive surface. Human peripheral corneal cells were maintained using a sphere‐forming assay and their behaviour on collagen substrate recorded by time‐lapse imaging. Immunocytochemistry and proliferation assays were used to detect protein expression and cell division. Proliferation assays showed that spheres formed by a combination of cell division and aggregation. Cell division continued within spheres for up to 4 months and was up‐regulated when exposed to differentiation medium and collagen substrate. The spheres expressed both epithelial and stromal cell markers. When exposed to collagen; (1) 25% of the spheres showed spontaneous polarised outgrowth. (2) One sphere initially showed polarised outgrowth followed by collective migration with discrete morphological changes to form leading and trailing compartments. (3) A sphere which did not show polarised outgrowth was also capable of collective migration using cell protrusion and retraction. (4) Active recruitment of cells into spheres was observed. (5) Placement of spheres in close proximity led to production of a cell exclusion area adjacent to spheres. Thus peripheral corneal cell spheres are dynamic entities capable of developing polarity and modifying migration in response to their environment.