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Biomedical subjects

Sheng Ding

Publications and source records attributed to Sheng Ding.

At least 19 recordsLinked to original sources
PubMed2006

Self-renewal of embryonic stem cells by a small molecule.

A cell-based screen of chemical libraries was carried out to identify small molecules that control the self-renewal of ES cells. A previously uncharacterized heterocycle, SC1, was discovered that allows one to propagate murine ES cells in an undifferentiated, pluripotent state under chemically defined conditions in the absence of feeder cells, serum, and leukemia inhibitory factor. Long-term SC1-expanded murine ES cells can be differentiated into cells of the three primary germ layers in vitro and also can generate chimeric mice and contribute to the germ line in vivo. Biochemical and cellular experiments suggest that SC1 works through dual inhibition of RasGAP and ERK1. Molecules of this kind may not only facilitate practical applications of stem cells in research and therapy, but also provide previously undescribed insights into the complex biology of stem cells.

Animals↗
PubMed2006

Integrin-linked kinase regulates Bergmann glial differentiation during cerebellar development.

We demonstrate here that integrin-linked kinase (ILK), a serine/threonine kinase that binds to the beta1 integrin cytoplasmic domain, regulates cerebellar development. Mice with a CNS-restricted knock-out of the Ilk gene show perturbations in the laminar structure of the cerebellar cortex that are associated with defects in Bergmann glial fibers and the formation of meningeal basement membranes. Similar defects have been observed in mice lacking beta1 integrins in the CNS. ILK and beta1 integrins are coexpressed in Bergmann glial cells, and studies with primary cells in culture demonstrate that ILK and CDC42 are required for beta1-integrin-dependent glial process outgrowth. Consistent with these findings, the amount of GTP-bound CDC42 is impaired in the cerebellum of Ilk-deficient mice. We conclude that beta1 integrin, ILK and CDC42 are components of the signaling machinery that regulates glial process outgrowth in the cerebellum. We also show that granule cell precursor proliferation is affected in ILK-deficient mice, but our findings provide strong evidence that proliferative defects are a secondary consequence of ILK function in glia.

Actins↗
PubMed2006

Long-term self-renewal and directed differentiation of human embryonic stem cells in chemically defined conditions.

Chemically defined medium (CDM) conditions for controlling human embryonic stem cell (hESC) fate will not only facilitate the practical application of hESCs in research and therapy but also provide an excellent system for studying the molecular mechanisms underlying self-renewal and differentiation, without the multiple unknown and variable factors associated with feeder cells and serum. Here we report a simple CDM that supports efficient self-renewal of hESCs grown on a Matrigel-coated surface over multiple passages. Expanded hESCs under such conditions maintain expression of multiple hESC-specific markers, retain the characteristic hESC morphology, possess a normal karyotype in vitro, as well as develop teratomas in vivo. Additionally, several growth factors were found to selectively induce monolayer differentiation of hESC cultures toward neural, definitive endoderm/pancreatic and early cardiac muscle cells, respectively, in our CDM conditions. Therefore, this CDM condition provides a basic platform for further characterization of hESC self-renewal and directed differentiation, as well as the development of novel therapies.

Activins↗
PubMed2006

Allosteric inhibitors of Bcr-abl-dependent cell proliferation.

Chronic myelogenous leukemia (CML) is a myeloproliferative disorder characterized at the molecular level by the expression of Bcr-abl, a 210-kDa fusion protein with deregulated tyrosine kinase activity. Encouraged by the clinical validation of Bcr-abl as the target for the treatment of CML by imatinib, we sought to identify pharmacological agents that could target this kinase by a distinct mechanism. We report the discovery of a new class of Bcr-abl inhibitors using an unbiased differential cytotoxicity screen of a combinatorial kinase-directed heterocycle library. Compounds in this class (exemplified by GNF-2) show exclusive antiproliferative activity toward Bcr-abl-transformed cells, with potencies similar to imatinib, while showing no inhibition of the kinase activity of full-length or catalytic domain of c-abl. We propose that this new class of compounds inhibits Bcr-abl kinase activity through an allosteric non-ATP competitive mechanism.

Animals↗
PubMed2006

Conformational changes and inactivation of bovine carbonic anhydrase II in 2,2,2-trifluoroethanol solutions.

Changes in unfolding and enzymatic activity of bovine carbonic anhydrase II (BCA II) in different concentrations of 2,2,2-trifluoroethanol (TFE) were investigated by 1-anilino-8-naphthalenesulfonate (ANS) fluorescence emission spectra, far-UV CD spectra, and enzyme activity. The results showed that the activity and conformation of BCA II changed according to the concentration of TFE. Significant aggregation was observed when BCA II was denatured at TFE concentrations between 10 and 35% (v/v). When the concentration of TFE exceeded 40%, the aggregation of BCA II was not very obvious. The activity of BCA II decreased almost to zero as the TFE concentration reached 26%. The ANS fluorescence spectra indicated the tertiary conformations of BCA II were more stable in solutions with TFE concentrations lower than 15% (v/v) and higher than 40% (v/v). Far-UV CD spectra showed that high concentrations (higher than 25%) of TFE could induce BCA II to form more alpha-helix structures and caused these structures to be in relatively stable states. The native conformation of BCA II being destroyed after its inactivity indicated that the active sites of BCA II is situated in a limited region and has more flexibility than the whole enzyme molecule.

Animals↗
PubMed2006

Generation of RNAi libraries for high-throughput screens.

The completion of the genome sequencing for several organisms has created a great demand for genomic tools that can systematically analyze the growing wealth of data. In contrast to the classical reverse genetics approach of creating specific knockout cell lines or animals that is time-consuming and expensive, RNA-mediated interference (RNAi) has emerged as a fast, simple, and cost-effective technique for gene knockdown in large scale. Since its discovery as a gene silencing response to double-stranded RNA (dsRNA) with homology to endogenous genes in Caenorhabditis elegans (C elegans), RNAi technology has been adapted to various high-throughput screens (HTS) for genome-wide loss-of-function (LOF) analysis. Biochemical insights into the endogenous mechanism of RNAi have led to advances in RNAi methodology including RNAi molecule synthesis, delivery, and sequence design. In this article, we will briefly review these various RNAi library designs and discuss the benefits and drawbacks of each library strategy.

Journal Article↗
PubMed2006

[Application of micronucleus test of buccal mucosal cells in assessing the genetic damage of workers exposed to acrylonitrile].

OBJECTIVE: To investigate the application of micronucleus test of buccal mucosal cells in monitoring the genetic effect of acrylonitrile in the population exposed to the acrylonitrile. METHODS: Forty-one healthy male workers in a chemical factory in Shanghai were selected as the low concentration acrylonitrile exposed group while forty-seven healthy male workers in an acrylonitrile factory in Shanghai were selected as the intermediate concentration acrylonitrile exposed group. At the same time, thirty-one male workers who had no toxicant exposure and lived in the same community were selected as the control group. The micronucleus test in buccal mucosal cells and lymphocytes were used respectively for assessing the genetic damage status of these men. RESULTS: The rate of micronucleus in buccal mucosal cells in both acrylonitrile groups (the low concentration group: 3.68% +/- 2.72%; the intermediate concentration group: 4.00% +/- 2.38%) was significantly higher than that in the control group (2.03% +/- 2.20%) (P < 0.05). The rate of micronucleus in the intermediate concentration group (4.23% +/- 3.34%) was significantly higher than that in the control group (2.48% +/- 1.46%) (P < 0.05). There was the correlation between the micronucleus test of buccal mucosal cells and the micronucleus test of the lymphocytes in the peripheral blood in the acrylonitrile exposed population (r = 0.299-0.359, P < 0.05). CONCLUSION: The micronucleus test of buccal mucosal cells replacing the micronucleus test of the lymphocytes in the peripheral blood can be used as one of the screening indexes in the surveillance of the genetic damage in the acrylonitrile exposed population.

Acrylonitrile↗
PubMed2005

Exploring stem cell biology with small molecules.

Stem cells hold promise for the treatment of a number of diseases. Small molecules serve as useful chemical tools to control stem cell fate and may ultimately contribute to development of effective medicines for tissue repair and regeneration.

Adult↗
PubMed2005

Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice.

Transposable elements have been routinely used for genetic manipulation in lower organisms, including generating transgenic animals and insertional mutagenesis. In contrast, the usage of transposons in mice and other vertebrate systems is still limited due to the lack of an efficient transposition system. We have tested the ability of piggyBac (PB), a DNA transposon from the cabbage looper moth Trichoplusia ni, to transpose in mammalian systems. We show that PB elements carrying multiple genes can efficiently transpose in human and mouse cell lines and also in mice. PB permits the expression of the marker genes it carried. During germline transposition, PB could excise precisely from original insertion sites and transpose into the mouse genome at diverse locations, preferably transcription units. These data provide a first and critical step toward a highly efficient transposon system for a variety of genetic manipulations including transgenesis and insertional mutagenesis in mice and other vertebrates.

Animals↗
PubMed2005

Identification of the Wnt signaling activator leucine-rich repeat in Flightless interaction protein 2 by a genome-wide functional analysis.

The Wnt signaling pathway acts ubiquitously in metazoans to control various aspects of embryonic development. Wnt ligands bind their receptors Frizzled and low-density lipoprotein receptor-related protein 5/6 and function through Disheveled (Dvl), Axin, adenomatous polyposis coli, glycogen synthase kinase 3beta, and casein kinase (CK) 1 to stabilize beta-catenin and induce lymphocyte enhancer-binding factor (LEF)/T cell factor (TCF)-dependent transcriptional activities. To identify previously unrecognized Wnt signaling modulators, a genome-wide functional screen was performed using large-scale arrayed cDNA collections. From this screen, both known components and previously uncharacterized regulators of this pathway were identified, including beta-catenin, Dvl1, Dvl3, Fbxw-1, Cul1, CK1epsilon, CK1delta, and gamma-catenin. In particular, a previously unrecognized activator, LRRFIP2 (leucine-rich repeat in Flightless interaction protein 2), was found that interacts with Dvl to increase the cellular levels of beta-catenin and activate beta-catenin/LEF/TCF-dependent transcriptional activity. The function of LRRFIP2 is blocked when a dominant negative Dvl (Xdd1) is coexpressed. Expression of LRRFIP2 in Xenopus embryos induced double axis formation and Wnt target gene expression; a dominant negative form of LRRFIP2 suppresses ectopic Wnt signaling in Xenopus embryos and partially inhibits endogenous dorsal axis formation. These data suggest that LRRFIP2 plays an important role in transducing Wnt signals.

Adaptor Proteins, Signal Transducing↗
PubMed2005

Small molecules and future regenerative medicine.

Recent advances in stem cell biology may make possible new approaches for the treatment of a number of diseases including cardiovascular disease, neurodegenerative disease, musculoskeletal disease, diabetes and cancer. These approaches could involve cell replacement therapy and/or drug treatment to stimulate the body's own regenerative capabilities by promoting survival, migration/homing, proliferation, and differentiation of endogenous stem/progenitor cells. However, such approaches will require identification of renewable cell sources of engraftable functional cells, an improved ability to manipulate their proliferation and differentiation, as well as a better understanding of the signaling pathways that control their fate. Cell-based phenotypic and pathway-specific screens of synthetic small molecules and natural products have historically provided useful chemical ligands to modulate and/or study complex cellular processes, and recently provided a number of small molecules that can be used to selectively regulate stem cell fate and developmental signaling pathways. Such molecules will likely provide new insights into stem cell biology, and may ultimately contribute to effective medicines for tissue repair and regeneration.

Animals↗
PubMed2005

Relations of perceived physical self-efficacy and motivational responses toward physical activity by urban high school students.

To be effective in promoting physical activity among urban, minority adolescents, the factors and psychological processes that motivate them to engage in and maintain a physically active lifestyle should be examined. The relation of physical self-efficacy and motivational responses toward physical activity in 46 urban minority adolescents was explored. As hypothesized, there were significant positive relationships among Percieved Physical Ability, Physical Self-presentation Confidence, Effort, and Enjoyment (coefficients ranged from .29 to .80), suggesting that participants who had higher perceived physical ability were likely to report higher perceptions of self-presentation, more enjoyment of physical activity, and harder work in physical activity. These results indicate specific relationships among Effort, Enjoyment, Perceived Physical Ability, and Physical Self-presentation Confidence in this sample. Physical self-efficacy appears to be a stronger predictor of motivational responses in physical activity. Practical implications for physical educators include incorporating strategies known to develop self-efficacy, such as mastery experiences involving successive trials of increasing difficulty, self-observation, external feedback, peer modeling, and verbal persuasion.

Adolescent↗
PubMed2005

Genomic studies in stem cell systems.

Stem cell research holds promise for future regenerative medicine. In recent years, much effort has been devoted to our understanding of stem cell self-renewal and differentiation at the molecular level. Important studies that have applied various genomic approaches, including genomics, functional genomics and proteomics, to gain molecular insights into stem cell development, are reviewed.

Expressed Sequence Tags↗
PubMed2005

[Investigation of the current situation on clinical application of endodontic treatments in Shanghai municipality].

PURPOSE: To investigate the present state of endodontic treatments applied in Shanghai municipality. METHODS: A questionnaire survey about clinic application of three endodontic treatments including pulp mummification (PM), resinifying therapy(RT) and root canal therapy(RCT) was conducted by 136 dentists of 42 hospitals selected from three different types of hospitals in Shanghai. The questionnaire concerned the proportion and the scale of three treatments used clinically , the indications, the evaluation of the curative effect and the prospective application in future. SAS6.04 statistical software was used for Chi square test and Kruskal-Wallis test. RESULTS: (1) PM and RCT were applied widely, RT was seldom used. There was significant difference in the proportion using PM among three different types of hospitals (P<0.001). (2) PM was primarily applied to the teeth suffering from acute or chronic pulpitis, especially molars with vital pulp.(3)Good long-term curative effect was the primary factor affecting dentists to chose endodontic treatment.(4) Most dentists affirm that the best curative effect would be obtained for RCT, however, the effect was uncertain for PM to treat acute or chronic pulpitis. CONCLUSION: The current situation of clinical use of endodontic therapeutic methods in Shanghai falls some short of the concepts accepted internationally. But in general, RCT should be popularized while PM should be eliminated gradually.

China↗
PubMed2004

Methylation profile of the promoter CpG islands of 14 "drug-resistance" genes in hepatocellular carcinoma.

AIM: To establish the DNA methylation patterns of the promoter CpG islands of 14 "drug-resistance" genes in hepatocellular carcinoma (HCC). METHODS: The methylation specific polymerase chain reaction in conjunction with sequencing verification was used to establish the methylation patterns of the 14 genes in the liver tissues of four healthy liver donors, as well as tumor and the paired non-cancerous tissues of 30 HCC patients. RESULTS: While 11 genes (ATP-binding cassette, sub-family G (WHITE), member 2(ABCG2), activating transcription factor (ATF2), beta-2-microglobulin (B2M), deoxycytidine kinase (DCK), occludin (OCLN), v-raf-1 murine leukemia viral oncogene homolog (RAF1), ralA binding protein 1 (RALBP1), splicing factor (45 kD) (SPF45), S-phase kinase-associated protein 2 (p45) (SKP2), tumor protein p53 (Li-Fraumeni syndrome) (TP53) and topoisomerase (DNA) II beta (TOP2B)) maintained the unmethylated patterns, three genes displayed to various extents the hypermethylation state in tumor tissues in comparison with the normal counterparts. The catalase (CAT) was hypermethylated in tumor and the neighboring non-cancerous tissue of one case (3.3%). Both glutathione S-transferase pi (GSTpi) (80%, 24/30 in tumor and 56.7%, 17/30 in the paired non-cancerous tissues) and cystic fibrosis transmembrane conductance regulator, ATP-binding cassette (sub-family C, member 7) (CFTR) (77%, 23/30 in tumor and 50%, 15/30 in the paired non-cancerous tissues) genes were prevalently hypermethylated in HCC as well as their neighboring non-cancerous tissues. No significant difference in the hypermethylation occurrence was observed between the HCC and its neighboring non-cancerous tissues. CONCLUSION: Hypermethylation of promoter CpG islands of both CFTR and GSTpi genes occurs prevalently in HCC, which may correlate with the low expression of these two genes at the mRNA level and has the profound etiological and clinical implications. It is likely to be specific to the early phase of HCC carcinogenesis.

Acyltransferases↗
PubMed2004

Small molecules that induce cardiomyogenesis in embryonic stem cells.

A phenotypic cell-based screen of a large combinatorial chemical library led to the identification of a class of diaminopyrimidine compounds (cardiogenol A-D) which can selectively and efficiently induce mouse embryonic stem cells (ESCs) to differentiate into cardiomyocytes. ESC-derived cardiomyocytes were shown to express multiple cardiac muscle markers, including myosin heavy chain, GATA-4, MEF2, and Nkx2.5, and spontaneously form beating regions. Such small molecules will serve as useful chemical probes to study cardiac muscle differentiation and may ultimately facilitate the therapeutic application of ESCs for cardiac repair.

Aniline Compounds↗