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Yuan-Liang Wang

Publications and source records attributed to Yuan-Liang Wang.

9 recordsLinked to original sources

The MGF expression of osteoblasts in response to mechanical overload.

Cell proliferation and mRNA expression of insulin-like growth factor (IGF-I) and "mechanogrowth factor" (MGF) were studied in osteoblasts in response to overload. Static and cyclic-stretching were used to apply superphysiological strains to cells. Overload was found to increase cell growth. IGF-I and its splicing variant, MGF, were measured using reverse transcriptase-polymerase chain reaction method and were found to be regulated differentially by mechanical signals at the mRNA level. Cyclic-stretching had a more significant effect on cell proliferation and mRNA expression levels of IGF-I and MGF, while unstrained cells did not express MGF at the mRNA level. These results demonstrated that gene expression is regulated by mechanical stimulation. MGF expression in osteoblasts in response to strain may be related to an autocrine mechanism.

Animals↗

[Phylogenetic diversity analyse of rumen bacteria using culture independent method].

Culture independent method was used to study the diversity of rumen bacteria. Molecular diversity of rumen bacteria was analyzed by PCR amplification and sequencing of 16S rDNA clone libraries prepared from the rumen content of Holstein cows. The total DNA directly extracted from rumen fluid was used as PCR template. Bacteria universal primer 27F and 1492R was used as primer. Random clones, containing almost full size 16S rDNA sequences (about 1.5 kb long), were sequenced and subjected to an on line similarity search. The 16S rDNA sequence analysis indicated that more than half of the sequences belonged to the not-yet-cultured groups. The 16S rDNA similarity levels with cultured species was less than 90%. The bacterial community structure was also revealed by phylogenetic tree of known sequences and selected sequence. In the library from the rumen fluid, the sequences were mainly affiliated with the following major phyla: low G + C Gram-positive bacteria, Cytophaga-Flexibacter-Bacteroides, and the remaining sequences were placed within not-yet-uncultured groups that had an uncertain affiliation. These several sequences are likely to represent novel taxonomic groupings. The nucleotide sequences have been submitted to the GenBank/EMBL /DDBJ databases under accession numbers AY986777-AY986791.

Animals↗

The stress reaction and its molecular events: splicing variants.

The growth of cells and tissues is regulated by stress. When body is injured, it manifests a large spectrum of metabolic, endocrine, and immune alterations, which is named stress reaction. Among them, the production of growth factors may play a critical role. For osteoblasts and myoblasts, IGF-I has been shown to be involved in the process of cells in response to overloads. There are two splicing forms, one is IGF-Ea, the other is the IGF-IEb in the rodents and corresponds to IGF-IEc in humans. The latter is markedly up-regulated in response to overloads. Therefore, it has been named mechanogrowth factor. The link between the mechanical stimulus and the gene expression represents a new and important area in cell science. Understanding the process of splicing in IGF-I helps one to investigate the mechanotransduction of cells in response to mechanical stimulation at molecular level.

Animals↗

The effect of step-wise increased stretching on rat calvarial osteoblast collagen production.

Mechanical forces regulate the function of bone cells. In this paper, the effects of cyclic stretching on osteoblasts derived from rat calvaria were studied at a magnitude occurring in physiological loaded bone tissue. A four-point bending apparatus was used to apply cyclic stretching on osteoblasts. Stretching at 500 microepsilon for 2-24 h resulted in an increase in matrix synthesis(P<0.01). In contrast, the cyclic stretching at 1000 and 1500 microepsilon for 2-24 h inhibited osteoblast collagen production (P<0.01). We also described our new loading method to increase strain magnitude step-by-step. The strain magnitude increased by 500 microepsilon increments from 500 to 1500 microepsilon every 2 or 12 h, respectively. Results showed that osteoblasts could absorb large amount of proline for collagen synthesis when stretched at 500 microepsilon. However, not all the absorbed proline was used to synthesize collagen. Some of it was stored in cells. When the suitable signal (500 microepsilon) was changed to an inhibiting signal (1000 microepsilon), cells responded to it accordingly and released proline to medium. These results demonstrate that the response of osteoblasts is dependent on the magnitude of the strain applied and cells can adjust their bio-chemical response to adapt to the changing environmental stimulation.

Animals↗

Preliminarily investigating the polymorphism of self-organized actin filament in vitro by atomic force microscope.

With the atomic force microscope (AFM), we preliminarily investigated the large-scale structure of actin filaments formed in low concentration protein solution (5 microg/ml) via self-organization without the presence of any F-actin dynamic interfering factors (such as phalloidin) in vitro. It was found that the G-actin could be polymerized into ordered filamentous structures with different diameter from the slimmest filament of single F-actin to giant filament in tree-like branched aggregates. The observed polymerized actin filaments, to which our most intense attention was attracted, was discretely distributed and showed obvious polymorphism distinctly different from those in the presence of phalloidin or actin binding proteins (fimbrin, gelsolin, etc.) in previous experiments. Latter structures were mainly composed of single F-actin and/or multifilaments clearly consisting of several single F-actin. The experimental results clearly demonstrated that non-interference with the F-actin intrinsic dynamics in self-organizing could lead to the polymorphism of actin filamentous structures, and further analysis implied that the disturbance of normal F-actin dynamics by many factors could prevent the emergence of structural polymorphism, more often than not, give rise to formation of specific structures instead and different interference would bring about various particular structures under certain conditions.

Actin Cytoskeleton↗

Cloning, characterization and functional expression of a new beta-D-fructofuranosidase (Os beta fruct2) cDNA from Oryza sativa.

A new cDNA (Os beta fruct2) encoding an acid beta-D-fructofuranosidase from rice has been cloned, sequenced and expressed in Pichia pastoris. The full-length cDNA is 2453 base pairs long and encodes a pre-pro-protein of 682 amino acids. The cDNA fragment coding for mature enzyme was sub-cloned into vector pPICZalphaA for extracellular expression in the methylotrophic yeast. The recombinant product was purified by Ni2+-nitrilotriacetic acid agarose column and biochemically characterized. The enzyme could hydrolyze sucrose and raffinose. Molecular mass is 66 kDa. The activity optimum was at pH 4.8 and 40 degrees C.

Amino Acid Sequence↗

Atomic force microscopy of actin.

Atomic force microscope (AFM) is a powerful and novel tool to investigate the surface and submembranous structures of living cells under physiological conditions at high resolution, and has force measure at nano-Newton level. AFM can image and manipulate samples (single macromolecule, cells, and so on) at very high, sometimes atomic resolution by scanning a fine tip over the surface of interest and detecting physical interactions between the tip and sample. Actin is one of the most important proteins, involved extensively in various cell physiological activities and functions in eukaryotes. The present paper clarified the crucial significance and the enormous applicable prospects of AFM in modern biological sciences by reviewing the usage of AFM in actin study, which can be taken as a good example for expounding almost all functions that AFM owns, and can be utilized in life sciences exploration.

Actins↗

[Effects of mechanical strain on the proliferation of pulmonary epithelial cells and reorganization of integrins].

Using a cyclic strain unit, the proliferation and reorganization of alpha(5), beta(1 ) integrins in human pulmonary epithelial A549 cells that underwent mechanical strain were studied. Flow cytometry revealed that cellular proliferative index reduced significantly after cells were subjected to 15% elongation at frequency 20 cycles/min or 40 cycles/min for 48 h. Confocal microscopy revealed that alpha(5), beta(1 ) integrins in human pulmonary epithelial A549 cells transferred from the apic al layer to the basal layer and created adhesion plaques after 24 h exposure to 15% elongation at frequency 40 cycles/min. It is concluded that alpha(5), beta(1 ) integrins in pulmonary epithelial cells play an important role in transducing mechanical stress into intracellular signals.

Cell Division↗