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Biomedical subjects

D Gan

Publications and source records attributed to D Gan.

12 recordsLinked to original sources
PubMed2001

Temperature-jump investigations of the kinetics of hydrogel nanoparticle volume phase transitions.

The dynamics of the deswelling and swelling processes in thermoresponsive poly-N-isopropylacrylamide (pNIPAm) hydrogel nanoparticles have been studied by using time-resolved transmittance measurements, in combination with a nanosecond laser-induced temperature-jump (T-jump) technique. A decrease in the solution transmittance associated with deswelling of the particles has been observed as the solution temperature traverses the volume phase transition temperature of the particles. Upon inducing the T-jump, the deswelling transition only occurs in a small percentage (<10%) of the particle volume, which was found to be a thin periphery layer of the particles. The particle deswelling occurs on the microsecond time scale, and as shown previously, the collapse time can be tuned via adding small amounts of hydrophobic component to the particle shell. In contrast, the reswelling of the particles was thermodynamically controlled by bath equilibration, and only small differences in particle reswelling kinetics were found due to sluggish heat dissipation (millisecond time scale) from the sample cell.

Acrylic Resins↗
PubMed2001

Interfacial nonradiative energy transfer in responsive core-shell hydrogel nanoparticles.

Fluorescently labeled core-shell latex particles composed mainly of the thermoresponsive polymer poly-N-isopropylacrylamide (p-NIPAm) have been synthesized such that an energy transfer donor (phenanthrene) and an energy transfer acceptor (anthracene) are covalently localized in the core and shell, respectively. When the thermally induced particle deswelling is interrogated by photon correlation spectroscopy (PCS), a continuous (non-first order) phase transition is observed. Conversely, when the nonradiative energy transfer (NRET) efficiency is used to probe the collapse of these same particles, the phase transition event is observed to occur over a much smaller temperature range and approaches first-order (discontinuous) behavior. Furthermore, core-shell particles with differing shell thicknesses display identical phase transition temperatures when PCS is used to monitor the transition, while NRET measurements show a clear increase in collapse temperature as the shell thickness is increased. These apparently contradictory results are discussed in terms of a radial phase coexistence that exists in the microgel particles, which arises from a similarly radial inhomogeneity in the cross-linker concentration. The prospects for the NRET technique as a molecular-scale probe of nanostructured microgels are also discussed.

Biocompatible Materials↗
PubMed2001

Tunable swelling kinetics in core--shell hydrogel nanoparticles.

Thermoresponsive, core--shell poly-N-isopropylacrylamide (p-NIPAm) nanoparticles (microgels) have been synthesized by seed and feed precipitation polymerization, and the influence of chemical differentiation between the core and shell polymers on the phase transition kinetics and thermodynamics has been examined. The results suggest that the core--shell architecture is a powerful one for the design of colloidal "smart gels" with tunable properties. To examine these materials, differential scanning calorimetry (DSC), (1)H NMR, and temperature-programmed photon correlation spectroscopy (TP-PCS) have been employed. These measurements show that the addition of small concentrations of a hydrophobic monomer (butyl methacrylate, BMA) into the particle shell produces large decreases in the rate of thermo-induced particle collapse. Conversely, these low levels of hydrophobic modification do not perturb the thermodynamics of the particle phase transition. When these results are examined in light of previous studies of macroscopic hydrogels, they suggest that the formation of a thin, stable skin layer at the particle exterior during the early stages of particle collapse is the rate limiting factor in particle deswelling. Finally, the hydrophobicity (BMA content) of the shell determines the magnitude of the hydrogel collapse rate, while the thickness of the BMA containing region does not impact the observed kinetics. Together, these results suggest that control over the kinetics of microgel deswelling events can be accomplished simply by modification of the particle periphery, and therefore do not require homogeneous modification of the entire polymer structure.

Journal Article↗
PubMed2001

Human keratinocytes respond to osmotic stress by p38 map kinase regulated induction of HSP70 and HSP27.

Human skin is exposed to an environment that varies in humidity from 100 to 0%, leading to seasonal variations in the condition of the skin. Exposure to a low humidity environment creates an osmotic gradient across the stratum corneum, which is known to modulate cutaneous barrier function. Heat shock proteins protect against stress-induced destabilization of proteins. We investigated whether osmotic shock (sorbitol) induced a heat shock protein response in normal human keratinocytes, and used heat shock as a positive control. Both heat shock and osmotic stress (200 and 300 mM sorbitol) clearly induced heat shock proteins 70 and 27 mRNA levels. The induction of heat shock protein 70 mRNA levels by osmotic stress peaked at 16 h and persisted until 24 h, whereas upregulation of heat shock protein 70 mRNA levels by heat peaked at 2 h and returned to baseline levels by 6 h. Sorbitol also increased heat shock protein 70 levels in a concentration-dependent manner. The kinetics of heat shock protein 27 mRNA induction by osmotic stress and heat were similar with peak induction at 6 h. The mitogen activated protein kinase family of proteins plays an important part in the coordination of gene responses to various stress conditions. We have demonstrated that the p38 mitogen activated protein kinase was strongly activated by 200 mM and 300 mM sorbitol. The specific p38 mitogen activated protein kinase inhibitor PD169316 almost completely blocked heat shock protein 70 mRNA induction by 200 mM and 300 mM sorbitol and completely suppressed heat shock protein 27 mRNA induction with 200 mM sorbitol. PD169316 also counteracted upregulation of heat shock protein 70 levels by sorbitol. These data indicate that keratinocytes respond to osmotic stress by p38 mitogen activated protein kinase regulated induction of heat shock proteins. This molecular pathway may be relevant for the mechanisms regulating the response of human skin to variations in environmental humidity.

Cells, Cultured↗
PubMed2000

Successful separation of apoptosis and necrosis pathways in HaCaT keratinocyte cells induced by UVB irradiation.

UVB irradiation can induce apoptotic, necrotic, and differentiation pathways in normal human keratinocytes. The present study was undertaken to determine at what dose of UVB each of these pathways is induced and whether these pathways are distinct or overlapping. We have observed that UVB induces fragmentation of DNA in human HaCaT keratinocytes, in a bimodal manner. Low doses of UVB, 5-20 mJ/cm2, increase the levels of apoptosis as shown by increased levels of fragmented DNA, Fas, PARP, and FasL protein, and the number of apoptotic cells as assessed by FACS analysis. At higher doses of UVB (20 and 30 mJ/cm2) the number of apoptotic cells becomes reduced, as does the amount of Fas, PARP, and FasL protein. At these higher doses, cell viability is decreased as measured by DNA synthesis (BrdU labeling) neutral red uptake, which represents an increasing necrotic phenotype. Expression of markers of keratinocyte differentiation, involucrin, keratin K1, and keratin K10, are also observed to decrease with increasing UVB dose. These changes are accompanied by a further increase in DNA fragmentation. We conclude that low doses of UVB (5-20 mJ/cm2) induced an apoptotic pathway, whereas increasing doses (greater than 20 mJ/cm2) of UVB produce a direct necrotic effect and inhibit terminal differentiation.

Apoptosis↗
PubMed1999

[The standardized techniques for preparing samples used in pathologic diagnostic transmission electron microscopy].

Transmission electron microscopy (TEM) has been used widespreadly in the field of diagnostic pathology. Based on the practical domestic condition and aimed at fully satisfying the needs of the pathologic diagnostic TEM, a set of standardized techniques were established for preparing different kinds of samples. The techniques included the preparation and storage of fixatives and embedding media, the requirement of sampling, the 5 h and 24 h embedding procedures for fresh specimens, the embedding procedure for suspension specimens and specimens collected from fine needle aspiration biopsy, as well as the procedures for preparing samples taken from paraffin embedded blocks or paraffin sections. In practical use, we found that the advantage of these techniques was that the ultrastructures of the tissues and cells were well preserved and they perfectly met the needs for diagnosis of tumors, kidney diseases, et al. The main points of the standardized techniques have been discussed in detail in this paper.

Breast Neoplasms↗
PubMed1999

[A method of preparing the section of eye tissue for transmission electron microscopy].

A method of preparing the section of eye tissue for transmission electron microscopy (TEM) is recommended. The tissue was prefixed with a mixed solution of 4% paraformaldehyde and 2.5% glutaraldehyde and followed by softening in 3% EDTA solution for 20 minutes, then the tissue was fixed in 1% osmium tetroxide, dehydrated in series acetone, infiltrated in Epox 812 for a longer, and embeded. Ultrathin sections were cut with glass knives or diamond knife, stained with uranyl acetate and lead citrate and examined with H600-IV. The advantage of this method is that the ultrastructures of tissues are well preserved without any damage and distortion. The main points of the procedure of preparation have been discussed.

Animals↗
PubMed1998

[Effects of smoking on interleukin-2 and its receptor system in human peripheral blood].

42 healthy male smokers and 42 healthy male nonsmokers matched in age and life style were strictly chosen into the study from Beijing urban and rural areas. Peripheral blood mononuclear cells(PBMCs) were separated and then stimulated by PHA under 37 degrees C, 5% CO2 condition. Immunochemical methods were applied in testing the synthesis of interleukin 2(IL-2) and the expression of its membrane receptor (mIL-2R) and the level of soluble receptor (sIL-2R) in serum. The results showed that when compared with nonsmokers, stimulated PBMC from smokers had significant higher percentage of mIL-2R positive cells (P < 0.05) in vitro. Levels of sIL-2R in serum of smokers were also significantly higher than those of nonsmokers (P < 0.05). The percentage of IL-2 secrete cells of smokers had higher tendency (P = 0.08). These data indicated that the immune function of smokers and nonsmokers are different concerning the IL-2 and its receptor system.

Adult↗
PubMed1992

[Observation of monoclonal antibody E4B7D5 inhibitory effect on leptospiral adherence using scanning electron microscope].

BALB/c mice were immunized intraperitoneally with outer envelopes of serogroup icterohaemorrhagiae lai serovar strain 017 leptospires. Monoclonal antibody (McAb) E4B7D5 against outer envelopes (IgG1, agglutinating titre 1:25,600) was produced by hybridoma technique. Passive immunoprotection experiments have demonstrated the immunoprotection of McAb E4B7D5 against strain 017 leptospires. Effect of McAb E4B7D5 on leptospiral adherence to the surface of normal human pulmonary embryonic fibroblasts was observed by using scanning electron microscope. The results indicated that the leptospiral adherence noted in various agglutinating titre McAb E4B7D5 groups was less frequent than that in the three control groups. It was concluded that the inhibitory effect of McAb E4B7D5 on leptospiral adherence may play a role in the immunoprotection.

Animals↗
PubMed1992

[Ultrastructural observation of SC1001-sodium on brain neurons kindled seizures by coriaria lactone in rabbits].

The ultrastructural changes of hippocampus and cerebral cortex neurons were derived from the rabbits kindled by coriaria lactone and from the kindled rabbits after treatment with SC1001 Na. The results of experiment showed that the hippocampus and cerebral cortex neurons in the kindled group had some degenerative changes, such as the swelling mitochondria, dilated rough endoplasmic reticulum and Golgi complexes when the injuries of neurons were not serious, but the seriously injured neurons were subjected to rupture of the cell membrane and solution of most organella, with only a small amount of degenerative organella. In the group treated with SC1001 Na, the ultrastructure of brain neurons demonstrated a tendency of recreation to normal which showed that SC1001 Na had cured many of the degenerative brain cells. This suggested that active treatment is very important for the epileptic patients.

Animals↗
PubMedDate not supplied

Normal human keratinocytes pretreated with damaged mRNA are resistant to UVB damage and exhibit increased DNA repair.

Treatment of normal human keratinocytes with UVC-irradiated rabbit globin mRNA 24 h before and after UVB exposure increased the survival of the human keratinocytes. We also observed that UVC-damaged mRNA reduced the formation of sunburn cells in skin models. We next tested the effects of UVC-damaged mRNA on cellular repair of DNA. DNA repair was evaluated using 2 assay methods. The first method used a damaged plasmid that is transfected back into the cell where it is repaired by the host cell repair mechanism. In these experiments, we observed that externally added UVC-damaged rabbit globin mRNA enhanced the repair of a plasmid transfected into the host keratinocyte cells. The second method used to determine the effects on DNA repair was direct immunostaining for thymidine-thymidine dimers (TT dimers) in histological sections of the skin models. Skin models were irradiated with UVB and then fixed immediately or after 24 h and stained for TT dimers. UVB irradiation immediately caused an increase in the number of stained keratinocytes in the skin. The number of stained cells decreased in skin fixed 24 h after UVB. This is due to repair of the TT dimers and their removal. Sections of skin models pretreated with UV-damaged mRNA exhibit greater removal of these TT dimers after 24 h. The above evidence suggests that damaged mRNA can trigger a host cell DNA repair pathway.

Animals↗