Biomedical subjects

C H Yao

Publications and source records attributed to C H Yao.

At least 19 recordsLinked to original sources

PubMed2002

Ginsenoside Rb1 enhances peripheral nerve regeneration across wide gaps in silicone rubber chambers.

Silicone rubber chambers filled with collagen containing ginsenoside Rb1 (GRb1) were used to repair lesioned rat sciatic nerves with 15-mm gaps between stumps. Six weeks after implantation, histology of the nerve regenerated in the chambers filled with GRb1 and collagen contained larger axons than those in the chambers with collagen only. This study showed that the GRb1 could exert a positive influence on nerve regeneration when using silicone rubber tubes.

Animals↗

PubMed2001

Effects of percutaneous electrical stimulation on peripheral nerve regeneration using silicone rubber chambers.

The purpose of this study was to determine whether 0.8-1 mA, 2 Hz of percutaneous electrical stimulation could affect the regeneration of a 10-mm gap of rat sciatic nerve created between the proximal and distal nerve stumps, which were sutured into silicone rubber tubes. Six weeks after implantation, though the group receiving the electrical stimulation had a lower success percentage of regeneration (57%) compared with the controls receiving no stimulation (70%), quantitative histology of the successfully regenerated nerves revealed that the mean values of the axon density, blood vessel number, blood vessel area, and percentage of blood vessel area in total nerve area in the group with the electrical stimulation were all significantly larger than those in the controls (p < 0.05). These results showed that the electrical stimulation could elicit rehabilitating effects on the regenerated nerves.

Animals↗

PubMed2001

A study on grafting and characterization of HMDI-modified calcium hydrogenphosphate.

It is known that the organic molecules can provide an effective means to manipulate the surface properties of the biodegradable ceramic. There are two ways to modify the surface of the biodegradable ceramic by organic molecules. The first one is through surface adsorption but organic molecules will easily be washed out in the physiological environment. The second approach is to graft organic molecules through covalent bond to the hydroxyl groups that are available on the surface of the ceramics. Isocyanate group has been reported as a coupling agent for hydroxyapatite and organic molecule. The studies showed that the isocyanate could react with hydroxyl groups of hydroxyapatite and form a covalent bond between isocyanate and hydroxyapatite. In the study, hexamethylene diisocyanate (HMDI) was used as coupling agent and calcium hydrogenphosphate (CaHPO4, CHP) was the candidate ceramic. CHP will react with HMDI at the temperature of 20 degrees C, 30 degrees C, 40 degrees C, 50 degrees C, 60 degrees C, and 70 degrees C for 4h. Dibutyltin dilaurate and hydroquinone were used as catalyst and inhibitor, respectively. The effect of reaction temperature on the grafted yield will be described. The linkage between CHP and HMDI will be characterized by DTA, TGA, FTIR, XRD, and 31P, 13C liquid state NMR. From the results, we successfully modified the surface of CHP with coupling agent of HMDI. The grafted yield of HMDI on CHP was increasing with the reaction temperature. The best temperature for CHP modified by HMDI is around 50 degrees C. The linkage between HMDI and the surface of CHP is a urethane linkage as CHP-O-CO-NH-(CH2)6-N=C=O. After further treatment, the terminal group of CHP treated with HMDI (MCHP) will be converted into a primary amine group as the formula of CHP-O-CO-NH-(CH2)6-NH2. If reaction temperature is 60 degrees C, long extension chain will occur with a urea linkage between the isocyanate groups as the formula of CHP-O-CO-NH-(CH2)6-(NH-CO-NH-(CH2)6)n-NH2. At reaction temperature higher than 60 degrees C, the HMDI will become prepolymerized forms in solution. The prepolymerized forms such as allophanate, biuret, uretidione and urea linkage will turn the solution into gel type mixture, which will lead to low grafted yield of HMDI on CHP. When MCHP prepared at the temperature 20 degrees C, there is no evidence of long extension but the grafted yield is the lowest only 0.9 wt% around.

Calcium Phosphates↗

PubMed2001

Osteogenic evaluation of glutaraldehyde crosslinked gelatin composite with fetal rat calvarial culture model.

The cytotoxicity of the synthetic bone substitute composed of tricalcium phosphate and glutaraldehyde crosslinked gelatin (GTG) were evaluated by osteoblast cell culture. In a previous study, the GTG composites were soaked in distilled water for 1, 2, 4, 7, 14, 28, and 42 days, and then the solutions (or extracts) were cocultured with osteoblasts to evaluate the cytotoxicity of GTG composites by alive cell counting. In this study, the extracts were cocultured with the osteoblasts; thereafter, the concentration of transforming growth factor-beta (TGF-beta1) and prostaglandin E2 (PGE2) in the medium was analyzed to strictly reflect the biological effects of GTG composites on the growth of osteoblasts. In order to investigate the osteoconductive potential of the GTG composites on new bone formation in a relative short term, a model of neonatal rat calvarial organ culture was designed prior to animal experiments. Three experimental materials of 4, 8, and 12% GTG composites were evaluated by fetal rat calvarial organ culture for their ability for bone regeneration. Deproteinized bovine and porcine cancellous bone matrixes were used as the controlled materials. All the organ culture units were maintained in cultured medium for 5 weeks. Following the culture period, the morphology of tissue was observed under an optical microscope, and the quantitative evaluation of the new generation bone was determined by using a semiautomatic histomorphometeric method. Except in the initial 4 days, the concentration of TGF-beta1 of 4% and 8% GTG composites was higher than that of the blank group for all the other experimental time periods. The PGE2 concentration for 4% and 8% GTG composites was lower than that of the blank group. It revealed that the 4% and 8% GTG composites would not lead to inflammation and would promote osteoblast growth. The morphology and activity of the osteoblasts were not transformed or changed by the 2 GTG composites. For the 12% GTG composite, the performance of the in vitro condition was inferior to the blank group and the other 2 GTG composites. Although the concentration of TGF-beta1 and PGE2 was gradually back to normal after 14 days, the morphology of the osteoblasts was abnormal with features such as contracted cytoplast structures. The osteoblast was damaged perhaps in the initial stage. We suggested that the 4% and 8% GTG composites should be soaked in distilled water at least for 4 days before medical applications. The 12% GTG composite and the composites with a concentration of glutaraldehyde solution higher than 12% were not recommended as a medical prostheses in any condition. The fetal rat calvaria culture also showed the same results with the analysis of TGF-beta1 and PGE2. From the study, we could predict the results of animal experiments in the future.

Animals↗

PubMed2001

Effect of acupuncture stimulation on peripheral nerve regeneration using silicone rubber chambers.

The purpose of this study was to determine whether acupuncture could affect the regeneration of a 10-mm gap of rat sciatic nerve created between the proximal and distal nerve stumps, which were sutured into silicone rubber tubes. Empty silicone rubber tubes with no further treatment were used as controls. Six weeks after implantation, the animals received the acupuncture or the electroneedling treatment exhibited a more mature ultrastructural nerve organization with significantly higher numbers in the axon density, the blood vessel area, and the percentage of blood vessel area occupied in total nerve area than the controls. In addition, the electroneedling could combine both the needling and the electrical stimulation to potentiate the nerve-growth promoting effect of the acupuncture treatment. These results showed that acupuncture treatment could elicit positive effects on regenerated peripheral nerves.

Acupuncture↗

PubMed2001

Effects of buyang huanwu decoction on peripheral nerve regeneration using silicone rubber chambers.

This study investigated the effect of buyang huanwu decoction on the regeneration of a 10-mm gap of rat sciatic nerve created between the proximal and distal nerve stumps, which were sutured into silicone rubber tubes. Empty silicone rubber tubes with no further treatment were used as controls. Six weeks after implantation, 89% of the animals orally administered the buyang huanwu decoction exhibited regeneration across the nerve gaps, whereas only 70% had regenerated in the control group. Both qualitative and quantitative histology of the regenerated nerves revealed a more mature ultrastructural organization with significantly higher numbers of myelinated axons, larger endoneurial areas, higher axon densities and a larger percentage of axon area per total nerve area in the buyang huanwu group than in the controls. These results showed that the buyang huanwu decoction had a growth-promoting effect on the regenerated nerves.

Animals↗

PubMed2000

Development of biodegradable polyesterurethane membranes with different surface morphologies for the culture of osteoblasts.

To evaluate the biocompatibility of biodegradable polyesterurethane membranes with different surface morphologies for their possible use as orthopedic biomaterials, rat osteoblasts were cultured on smooth, sunken, and particulate polyesterurethane membranes. A close interaction between cells and exposed particles on the particulate membranes was found. Cells on the particulate surfaces were well spread and flattened and had the greatest adhesion while cells on the smooth surfaces were more rounded, less spread, and less adhered. In addition, in order to investigate their in vivo degradation rates, the morphologic changes in retrieved membranes from 2, 4, and 8 weeks after subcutaneous implantation were observed by scanning electron microscopy and their average molecular weight changes were determined by gel permeation chromatography. These analyses showed that smooth membranes, compared with the two other surface membrane types, had the greatest rate and degree of molecular weight change. In contrast, the molecular weight of particulate membranes, which favor the osteoblast culture, had not changed significantly at 8 weeks postimplantation. Thus particulate polyesterurethane membrane surfaces may be of use as an orthopedic biomaterial, and polyesterurethane membranes certainly provide an ideal system for further study of the relative contributions to biocompatibility and degradation derived from surface morphology.

Animals↗

PubMed2000

Peripheral nerve regeneration using silicone rubber chambers filled with collagen, laminin and fibronectin.

A 10 mm gap of rat sciatic nerve was created between the proximal and distal nerve stumps, which were sutured into silicone rubber tubes filled with an extracellular gel containing collagen, laminin and fibronectin. Empty silicone rubber tubes were used as controls. Six weeks after implantation, all extracellular elements were completely degraded and absorbed, and 90% of the animals from the extracellular gel group exhibited regeneration across the nerve gaps, whereas only 60% in the control group. Both qualitative and quantitative histology of the regenerated nerves revealed a more mature ultrastructural organization with 28% larger cross-sectional area and 28% higher number of myelinated axons in the extracellular gel group than the controls. These results showed that the gel mixture of collagen, laminin and fibronectin could offer a suitable growth medium for the regeneration of axons.

Action Potentials↗

PubMed2000

Preparation and characterization of surface-modified calcium hydrogenphosphate by hexamethylene diisocyanates.

Isocyanate group has been reported as a coupling agent of hydroxyapatite and polymers. The studies showed that the isocyanate would react with hydroxyl groups of hydroxyapatite and form a covalent bond between isocyanate and hydroxyapatite. In the study, hexamethlene diisocyanate (HMDI) was used as coupling agent. Calcium hydrogen-phosphate (CaHPO4, CHP) powders was the candidate ceramic due to higher content of hydroxyl group, which would react with HMDI at the temperature of 30, 40, 50, 60, and 70 degrees C for 4 hours. Dibutyltin dilaurate and hydroquinone were used as catalyst and inhibitor, respectively. The product was analyzed by DTA, TGA, FTIR, XRD, 13C solid state NMR and 31P, 13C liquid state NMR. From the results, we could prove the surface of calcium hydrogen-phosphate has been successfully modified. The largest amount (5.6 wt%) of HMDI could be grafted on the surface of CHP when reacted at 50 degrees C for 4 hours. Some chain extension could be observed and their structure would also be described in the research.

Bone Substitutes↗

PubMed1999

Properties of the poly(vinyl alcohol)/chitosan blend and its effect on the culture of fibroblast in vitro.

In this work, the properties of poly(vinyl alcohol) (PVA) and PVA/chitosan blended membranes were investigated by scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and electron spectroscopy for chemical analysis (ESCA). The SEM photographs show the PVA/chitosan blended membrane undergoes dramatic changes on the surface and bulk structure during the membrane formation. The DSC analysis shows that PVA and chitosan are not very compatible in the PVA/chitosan blended membrane, whereas the combination of two polymer chains of constitutionally different features is revealed. In addition, the surface of the PVA/chitosan blended membrane is enriched with nitrogen atoms at the ESCA analysis. These reflect the PVA membrane can be modified by blending with chitosan that in turn may affect the biocompatibility of the blended membrane. Therefore, adhesion and growth of fibroblasts on the PVA as well as PVA/chitosan blended membranes were investigated. Cell morphologies on the membranes were examined by SEM and cell viability was studied using MTT assay. It was observed that the PVA/chitosan blended membrane was more favorable for the cell culture than the pure PVA membrane. Cells cultured on the PVA/chitosan blended membrane had good spreading, cytoplasm webbing and flattening and were more compacting than on the pure PVA membrane. Consequently, the PVA/chitosan blended membrane may spatially mediate cellular response that can promote cell attachment and growth, indicating the PVA/chitosan blended membrane should be useful as a biomaterial for cell culture.

Biocompatible Materials↗

PubMed1998

Biological effects and cytotoxicity of the composite composed by tricalcium phosphate and glutaraldehyde cross-linked gelatin.

The purpose of this study was to prepare and evaluate the feasibility and cytocompatibility of a composite (GTG) as a large defect bone substitute. The composite is combined with tricalcium phosphate ceramic particles and glutaraldehyde cross-linked gelatin. Gelatin had been reported as an adhesive and biocompatible binder that could accelerate the recovery of damaged soft tissue, but the effects of gelatin when acting on the bone tissue is not clear. Thus, it is necessary to determine if the substances released from the GTG composite can facilitate the growth of bone cells. The substances released from the GTG composites after being soaked in deionized distilled water were analyzed by gas chromatography (GC), ultraviolet and visible absorption spectroscopy (UV-VIS), and inductive-coupled plasma-atomic emission spectrometry (ICP-AES). The cytotoxicity of the GTG composites was assessed by coculture of rat osteoblasts in vitro. Extracts were obtained by soaking the GTG composites in deionized distilled water for 1, 2, 4, 7, 14, 28 and 42 d. The extract mixed with complete medium in a ratio of 1:1 was added into the cell culture wells containing 1 x 10(4) cells ml(-1) osteoblasts. After culturing for 2 days, the cells attached to the surface of wells were trypsinized and the number calculated by the Neubauer counting-chamber under the optical microscope. Finally, three samples in each GTG group were examined by scanning electron microscopy (SEM) to observe the morphology of the osteoblasts attached to the surfaces of GTG composites. The examinations of osteoblasts cocultured with the developed GTG composites were used to decide the ideal concentration of glutaraldehyde as a cross-linking agent. The results of extracts cocultured with osteoblasts showed that the extracts obtained from the 2, 4 and 8% glutaraldehyde cross-linked GTG composites would inhibit the growth of osteoblasts in the first 4 soaking days. During the 4-7 days soaking, the cell numbers quickly increased with the soaking time, thereafter, the cell numbers almost reached a constant value. In the analyses of substances released from the GTG composites, it was found that the gelatin and calcium were gradually released from the GTG composites, which were supposed to be nutritious for the growth of the osteoblast. The results of osteoblasts cocultured with the GTG composites showed that the concentration of glutaraldehyde used as a cross-linking agent should be lower than 8%. Compared to the GTF (composite combined with tricalcium phosphate ceramic particles and formaldehyde cross-linked gelatin), GTG composites were much suitable for a large defect bone substitute in the near future.

Animals↗

PubMed1997

Effects of calcium phosphate bioceramics on skeletal muscle cells.

With advances in ceramics technology, calcium phosphate bioceramics have been applied as bone substitutes. The effects of implants on bony tissue have been investigated. The effects upon adjacent skeletal muscles have not been determined. The focus of this work is to elucidate the biological effects of various calcium phosphate bioceramics on skeletal muscles. Four different kinds of powder of calcium phosphate biomaterials including beta-tricalcium phosphate (beta-TCP), hydroxyapatite (HA), beta-dicalcium pyrophosphate (beta-DCP) and sintered beta-dicalcium pyrophosphate (SDCP), were tested by myoblast cell cultures. The results were analyzed by cell count, cell morphology and concentration of transforming growth factor beta 1 (TGF-beta 1) in culture medium. The cell population and TGF-beta 1 concentration of the control sample increased persistently as the time of culture increased. The changes in cell population and TGF-beta 1 concentration in culture medium of the beta-TCP and HA were quite low in the first 3 days of culture, then increased gradually toward the seventh day. The changes in cell population and TGF-beta 1 concentration in culture medium of the silica, beta-DCP, and SDCP were quite similar. They were lower during the first day of culture but increased and reached that of the control medium after 7 days' culture. Most cells on B-TCP and HA diminished in size with radially spread, long pseudopods. We conclude that HA and beta-TCP are thought to have an inhibitory effect on growth of the myoblasts. The HA and beta-TCP may interfere with the repair and regeneration of injured skeletal muscle after orthopedic surgery.

Animals↗

PubMed1997

Regulatory sequences for the amplification and replication of the ribosomal DNA minichromosome in Tetrahymena thermophila.

We have analyzed the cis-acting sequences that regulate rRNA gene (rDNA) replication in Tetrahymena thermophila. The macronucleus of this ciliated protozoan contains 9,000 copies of a 21-kbp minichromosome in the form of a palindrome comprising two copies of the rDNA. These are derived from a single chromosomally integrated copy during conjugation through selective amplification and are maintained by replicating once per cell cycle during vegetative growth. We have developed a transformation vector and carried out a deletion analysis to determine the minimal sequences required for replication, amplification, and/or stable maintenance of the rDNA molecule. Using constructs containing progressively longer deletions, we show that only a small portion (approximately 900 bp) of the rDNA is needed for extrachromosomal replication and stable maintenance of this molecule. This core region is very near but does not include the rRNA transcription initiation site or its putative promoter, indicating that replication is not dependent on normal rRNA transcription. It includes two nearly identical nuclease-sensitive domains (D1 and D2), one of which (D1) corresponds to the physical origin of replication determined previously. Deletion of both domains abolishes replication, whereas deletion of either domain allows the molecules to replicate, indicating that only one domain is required. In addition to this core region, we have found several DNA segments, including a tandem array of a 21-nucleotide repeat (type II repeats) and sequences within the rRNA coding region, that play distinctive and important roles in maintaining the rDNA at a high copy number.

Animals↗

PubMed1994

Detection of circular excised DNA deletion elements in Tetrahymena thermophila during development.

Extensive programmed DNA deletion occurs in ciliates during development. In this study we examine the excised forms of two previously characterized deletion elements, the R- and M-element, in Tetrahymena. Using divergently oriented primers in polymerase chain reactions we have detected the junctions formed by joining the two ends of these elements, providing evidence for the presence of circular excised forms. These circular forms were detected in developing macronuclear DNA from 12-24 h after mating began, but not in micronuclear or whole cell DNA of vegetative cells. They are present at very low abundance, detectable after PCR only through hybridization with specific probes. Sequence analysis shows that the circle junctions occur at or very near the known ends of the elements. There is sequence microheterogeneity in these junctions, which does not support a simple reciprocal exchange model for DNA deletion. A model involving staggered cuts and variable mismatch repair is proposed to explain these results. This model also explains the sequence microheterogeneity previously detected among the junction sequences retained in the macronuclear chromosome.

Animals↗

PubMed1993

In situ hybridization at the electron microscopic level using a bromodeoxyuridine labeled DNA probe.

A bromodeoxyuridine (BrdU) labeled DNA probe was used for in situ hybridization at the electron microscopic (EM) level. A BrdU labeled DNA probe was hybridized in situ to cryostat sections of paraformaldehyde fixed OCT compound embedded cultured HL-60 cells. After hybridization, some sections were incubated with FITC-conjugated anti-BrdU monoclonal antibody for fluorescence microscopy (FM), and others were embedded in Quetol for electron microscopy (EM). The ultrathin sections of Quetol-embedded specimens were incubated with the anti-BrdU monoclonal antibody and the immunoglobulin: gold colloid. In both FM and EM studies, the signals were concentrated in the rough endoplasmic reticulum. Moreover, some label was arranged from the nucleus to the cytoplasm at the EM level. Relatively simple methods using the BrdU labeled DNA probe for the detection of the defined nucleic acid sequence with reasonable tissue preservation and high resolution are described here. This method may be useful for developmental and disease related studies of specific mRNA in cells and tissues.

Antibodies, Monoclonal↗

PubMed1992

Stroke in China (Sino-MONICA-Beijing study) 1984-1986.

We present here the results of the Sino-MONICA-Beijing stroke study based on 700,000 Beijing residents in 1984-1986. To compare incidence rates for stroke with other communities and countries, we adopted the criteria of the WHO Collaborative Study of 17 centers which used the same definition and methodology as was used in this study. Over the 3-year period of the study, 2,593 stroke events were registered in the 25- to 74-year age-group. The incidence rate for all strokes was 189.5/100,000 and the incidence rate for first strokes was 133.6/100,000. Men had a significantly higher incidence rate than women (all strokes 219.7/100,000 for male vs. 160.5/100,000 for female, OR = 1.32, 99% limits 1.19-1.46; first strokes 151.6/100,000 for male vs. 116.4/100,000 for female, OR = 1.25, 99% limits 1.11-1.42). In comparison with other studies, age-adjusted incidence rate of stroke in Beijing was higher than in other countries, especially for hemorrhagic stroke. The proportion of hemorrhagic stroke related to other types of stroke was also higher in Beijing. Further analysis of the cases confirmed by computerized tomography also supported this finding. Unlike the incidence rates, the 4-week case fatality rate for women, 39.5%, was higher than for men, 32.8%. This finding was confirmed by a multiple logistic analysis controlling for age (p less than 0.001) and for previous stroke (p less than 0.001). The adjusted sex difference is also significant (OR = 1.37, p less than 0.001). In addition, results showed that men had a higher hospitalization rate than women. More women than men were treated at home, possibly indicating better medical care for men.

Adult↗

PubMed1991

Anthropometric predictors of coronary heart disease and total mortality: findings from the US Railroad Study.

Prospective data from the US Railroad Study were used to investigate the relations of several anthropometric variables to coronary heart disease and all-cause mortality in males initially free of cardiovascular disease. Middle-aged men were examined in 1957-1960 and followed until 1977 or death. Anthropometric indicators of total body fat (body mass index and the sum of the subscapular and triceps skinfolds) and central body fat (the ratios chest circumference/biacromial (shoulder) diameter, and chest circumference/standing height) were significantly and directly associated with age-adjusted coronary heart disease mortality. When systolic blood pressure, serum cholesterol, cigarette smoking, and vital capacity were also taken into account, the ratio chest circumference/biacromial diameter remained significantly associated with coronary heart disease mortality. Total, central, and peripheral body fat had a "U"-shaped relation with all-cause mortality. Measures of the ratio of central to peripheral body fat were inconsistently related to mortality. These results indicate that certain anthropometric measurements, especially those that may indicate central adiposity, are positively related to the development of fatal coronary heart disease and quadratically related to all-causes death rates.

Adult↗

PubMed1991

Transformation of Tetrahymena to cycloheximide resistance with a ribosomal protein gene through sequence replacement.

A method for transforming Tetrahymena has been established earlier, but its application has been limited because of the lack of selectable markers other than the rRNA-encoding DNA (rDNA). Mutations in the yeast ribosomal protein L29 gene (CYH2) are known that confer cycloheximide resistance. We have cloned and sequenced the homologue of this gene from both a wild-type and a cycloheximide-resistant (ChxA) strain of Tetrahymena. Surprisingly, a comparison shows that the ChxA mutation is not present in the CYH2 homologue. We therefore created the yeast mutations in the Tetrahymena gene by site-directed mutagenesis and used them to transform Tetrahymena either with or without linking to an rDNA vector. All clones transformed by the rDNA vector also became resistant to cycloheximide when the rDNA contained the engineered mutant genes. Without the rDNA vector, the mutant genes transform approximately 1% of injected cells to become resistant to cycloheximide. DNA analysis indicates that transformation occurs by replacement of the host sequence and not by random integration of the injected sequence. The replacement occurs to some but not all copies of this gene in the polyploid macronuclear genome. Thus, transformation in Tetrahymena occurs by specific sequence replacement, and the injected mutant genes can serve as dominant selectable transformation markers in this organism.

Amino Acid Sequence↗