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Biomedical subjects

R He

Publications and source records attributed to R He.

At least 19 recordsLinked to original sources

The fluorescent characterization of the polymerized microtubule-associated protein Tau.

A new fluorescence formed while microtubule-associated protein tau was incubated at 25 and 37C for hours, with its maximum excitation at 230 and 280 nm, respectively. The fluorescence completely formed after tau was incubated in phosphate buffer and Tris-HCl buffer for approximately 20 h, with a relaxation phase about 2-4 h. The light scattering of the sample solution improved during formation of the fluorescence when tau was incubated. Both the fluorescence and tau oligomers did not form when tau was incubated in the buffers containing DTT. On the other hand, heparin improved both tau aggregation and the fluorescence formation. It suggests that the fluorescence comes from tau polymerization, which may follow the mechanism of tyrosine-tyrosinate emission for a protein not containing any tryptophan residues. This new fluorescence could be used as a probe to tau polymers.

Buffers↗

qBrain-2, a POU domain gene expressed in quail embryos.

We isolated a quail class III POU domain gene, qBrain-2, which was cloned from a cDNA library of E5 embryos. Northern blot and in situ hybridization analyses showed that qBrain-2 was expressed in developing central nervous system and adult brain. Moreover, qBrain-2 transcripts showed a dynamic distribution in embryonic central nervous system. Its transcripts were dominantly detected in the ventricular zone of the developing brain and spinal cord, but rarely in the differentiated region of mantle zone as well as the non-neuronal roof plate and floor plate. This suggests that qBrain-2 is involved in proliferation and differentiation of the neuroepithelial cells of quail central nervous system.

Amino Acid Sequence↗

Characterization of the effects of drug addition on the structure of glyceryl monoolein/water gel systems using low frequency dielectric spectroscopy.

The influence of propantheline bromide incorporation on the phase structure of glyceryl monoolein/water systems has been investigated using low-frequency dielectric spectroscopy over a frequency range of 10(-2) to 10(6) Hz at 20 degrees C. The responses of glyceryl monoolein systems composed of 10% and 30% w/w were measured and the spectra modeled using an equivalent circuit based on the Maxwell-Wagner theory.1,2 Marked changes in the dielectric responses of the systems were noted on addition of the propantheline bromide at concentrations up to 10% w/w. For the lamellar (10% w/w water) glyceryl monoolein systems, an increase in the imaginary permitivity was seen, corresponding to an increase in conductivity due to the presence of additional ionic species within the system. Evidence was also obtained for the incorporation of the drug directly into the lipid bilayers, particularly at higher concentrations (10% drug) at which dielectric behavior corresponding to bilayer disruption was seen. Incorporation of 3% and 5% w/w drug into the cubic phase systems (30% w/w water) resulted in a change to the lamellar phase. However, circuit modeling indicated that the system formed structures which showed features of both the lamellar and cubic phases at 3% w/w drug loadings. The study has therefore demonstrated that dielectric analysis may provide a novel means of studying the effects of drug incorporation on the phase behavior of complex gel systems.

Cholinergic Agents↗

An investigation into the supramolecular structure of ternary gel systems using oscillatory rheometry, microscopy, and low frequency dielectric spectroscopy.

A series of ternary gel systems based on cetostearyl alcohol (CSA) and cetomacrogol 1000 or sodium lauryl sulfate have been studied using oscillatory rheology, differential interference contrast (DIC) microscopy, cryoscanning electron microscopy (cryo-SEM), and low-frequency dielectric analysis in order to elucidate the nature of the lamellar structures formed in relation to composition. The effects of altering the concentration of CSA (0.25% to 8% w/w) for 1% and 2% w/v cetomacrogol 1000 and 0.5% and 1% w/v sodium lauryl sulfate systems have been investigated, with marked increases in the storage and loss moduli seen on increasing the concentration of CSA for both surfactants. DIC microscopy indicated that at low CSA concentrations, needlelike structures were seen which, on increasing the concentration, were observed to congregate into nuclei. At concentrations of 4% CSA and above, neospherical structures were also observed. Cryo-SEM revealed that the needlelike objects were sheet structures ascribed to lamellar gel phases, while the nuclei were folded "rosettes" formed by those sheets, with the spherical structures being ascribed to cetostearyl alcohol. It was also noted that the lamellae were more tightly folded at 8% w/w CSA, which may be associated with the higher rheological moduli for these systems. Low-frequency dielectric analysis was performed over a frequency range of 10(4) Hz to 10(-2) Hz. A decrease in both the dielectric loss and capacitance was observed as the concentration of cetostearyl alcohol was increased. The dielectric data were described in terms of an equivalent circuit model based on a modified Maxwell-Wagner response. A good correlation was found between the fitted and experimental data and the effect of altering the gel composition on specific features of the equivalent circuit are discussed.

Emulsions↗

PrM- and cell-binding domains of the dengue virus E protein.

The E-prM proteins of flaviviruses are unusual complexes which play important roles in virus assembly and fusion modulation and in potential immunity-inducing vaccines. Despite their importance, little is known about the biogenesis and structural organization of E-prM complexes. Pulse-chase radiolabeling of dengue virus-infected Vero cells demonstrated a rapid interassociation of E and prM proteins, and sucrose gradient sedimentation analysis suggested that E-prM complexes progressed from simple heteromers to more densely sedimenting structures indicating increased multimerization. E-prM heteromers of even higher complexity were observed in virus particles, suggesting an intracellular assembly process which results in the networking of E-prM subunits into a lattice-like structure found in virus particles. Trypsin cleavage of E-prM-containing virus particles resulted in the release of a soluble 45-kDa fragment of the E protein which retained cell-binding activity. The results suggest that E-prM interactions in dengue virus particles are largely mediated by domains in the carboxy-terminal anchoring domain of E, while cell-binding activity is retained in a trypsin-releasable ectodomain of the E protein.

Animals↗

Selection of quantitative criteria for evaluating metastases of a highly metastatic salivary adenoid cystic carcinoma cell line to the lungs of nude mice.

OBJECTIVE: To select reliable quantitative criteria for evaluating metastases of a highly metastatic salivary adenoid cystic carcinoma cell line (ACC-M) cell to the lungs of nude mice. MATERIALS AND METHODS: The within-group coefficients of correlation of 5 parameters were measured from lung slides using a video image digital analysis system (VIDAS), and the intergroup coefficients of correlation of 6 sets of parameters from all the lungs, which were taken from ACC-M lung metastases models, were calculated and statistically analyzed. RESULTS: The within-group coefficients of correlation of 5 parameters are as follows: ratio of area of metastases, t = 0.9 (P < 0.001); area of metastases, t = 0.7 (P < 0.05), size of lung section, numbers of metastases, and size of the largest metastatic focus all t < 0.4 (P > 0.05). The intergroup coefficients of correlation of the parameters are as follows: ratio of area of metastases was assigned 1, weight of the lung was 0.9755, and area of metastases was 0.8628. CONCLUSION: For evaluating lung metastases of ACC-M, the ratio of area of metastases and weight of the lung are precise; the area of metastases is also significant within the 6 quantitative parameters.

Animals↗

Enhanced efficacy of the combination of all-trans-retinoic acid and gamma-interferon in inhibiting proliferation of Tca8113 cells.

OBJECTIVE: To develop a more effective induced-differentiation or -apoptosis therapy for treating oral squamous cell carcinomas. METHODS: The combination of all trans retinoic acid (ATRA) and interferon-gamma (IFN-gamma) was tested for its antiproliferative activity against human tongue squamous cell carcinoma cell line (Tca8113 cells) in vitro. Clonogenic assay, MTT assay, and microscopic cell analysis were performed in this experiment. RESULTS: The results show that the combination of ATRA and IFN-gamma can produce synergistic antitumor effects, resulting from induction of apoptosis, on Tca8113 cells. CONCLUSION: The combination of ATRA and IFN-gamma is a more effective induced-apoptosis therapy for treating oral squamous cell carcinoma.

Apoptosis↗

Expression and characterization of deletion recombinants of two cGMP-inhibited cyclic nucleotide phosphodiesterases (PDE-3).

cDNAs encoding two PDE-3 or cyclic GMP-inhibited (cGI) cyclic nucleotide phosphodiesterase (PDE) isoforms, RPDE-3B (RcGIP1) and HPDE-3A (HcGIP2), were cloned from rat (R) adipose tissue and human (H) heart cDNA libraries. Deletion and N- and C-terminal truncation mutants were expressed in Escherichia coli in order to define their catalytic core. Active mutants of both RPDE-3B and HPDE-3A included the domain conserved among all PDEs plus additional upstream and downstream sequences. An RPDE-3B mutant consisting of the conserved domain alone and one from which the RPDE-3B 44-amino acid insertion was deleted exhibited little or no activity. All active recombinants exhibited a high affinity (< 1 microM) for cyclic AMP (cAMP) and cyclic GMP (cGMP), were inhibited by cAMP, cGMP, and cilostamide, but not by rolipram, and were photolabeled with [32P]-cGMP. The IC50 values for cGMP inhibition of cAMP hydrolysis were lower for HPDE-3A than for RPDE-3B recombinants. The deduced amino acid sequences of HPDE-3A and RPDE-3B catalytic domains are very similar except for the 44-amino acid insertion not found in other PDEs. It is possible that this insertion may not only distinguish PDE-3 catalytic domains from other PDEs and identify catalytic domains of PDE-3 subfamilies or conserved members of the PDE-3 gene family, but may also be involved in the regulation of sensitivity of PDE-3s to cGMP.

3',5'-Cyclic-AMP Phosphodiesterases↗

An investigation into the use of low-frequency dielectric spectroscopy as a means of characterizing the structure of creams based on Aqueous Cream BP.

A range of creams based on Aqueous Cream BP have been analyzed using low-frequency dielectric spectroscopy, with accompanying circuit modeling in combination with rheological and microscopic supportive techniques, to explore the use of the dielectric approach as a novel means of characterizing cream systems. Creams based on the formula for Aqueous Cream BP were produced by hand-mixing and mechanical mixing, with and without the inclusion of the preservative phenoxyethanol. Dielectric analysis was performed over a frequency range of 10(-2)-10(5) Hz. Cream samples were also examined using stress scan rheology and differential interference contrast microscopy. Dielectric analysis indicated that the presence of preservative decreased the capacitance and loss of the creams. The responses were modeled in terms of a dispersive capacitance in series with two RC circuits (series and parallel). Rheological studies indicated higher viscosities for the hand-mixed and unpreserved systems. Differential interference contrast microscopy showed marked differences in the distribution of the oil droplets, depending on the method of mixing. The study has demonstrated that dielectric spectroscopy, with accompanying circuit analysis, may be used as a means of modeling the structure of cream systems. The investigation has also shown that the formulation and preparation method of Aqueous Cream BP may have a profound effect on sample structure.

Chemistry, Pharmaceutical↗

Kinetics of selenium incorporation into tissues of female mallard ducks.

Selenium is essential for both mammalian and avian species, although its metabolism in birds has been less thoroughly studied. Little information has been available on the kinetics of selenium in birds, especially as it relates to the teratogenicity seen in waterfowl consuming excessive amounts. This study describes the pharmacokinetics of small amounts of 75Se as selenious acid injected into female mallard ducks. Labeled selenium was injected into a wing vein of restrained animals and tissues taken at five different time points up to 24 h post-injection. Selenium levels as percent of injected dose were determined in liver, kidney, heart, lung, adrenals, thyroid, spleen, pancreas, ovaries, intestine, muscle and plasma. Estimates of kinetic parameters (uptake and elimination rates, time of maximum concentration and maximum concentration) were obtained with a non-linear kinetics computer program (PCNONLIN, SCI Software, Lexington, KY). Results showed four basic patterns of distribution, uptake and elimination. Visceral tissues exhibited a triphasic pattern with a rapid rise, a decline followed by a distinctive increase in levels between the last two time points. Brain uptake was continuous over the 24 h. Plasma rose rapidly and then declined to a constant level. The ovaries as a tissue of interest relating to the teratogenic effects of selenium showed the greatest relative increase at 24 h, suggesting kinetic mechanisms consistent with a pathway that might lead to accumulation of toxic levels and teratogenic effects during embryo development.

Animals↗

Effect of thiols on fructosamine assay.

Fresh human serum, gGAPDH and beta-mercaptoethanol were used to examine the effect of thiols on fructosamine assay. The kinetics of the reaction with both fresh human serum and gGAPDH displayed biphasic behaviour (fast and slow). When the thiols were modified with IAA, the kinetics only demonstrated the slow phase. Since the absorbance increase in the interval from 9 to 10 min was used in the fructosamine assay of glycated human serum we studied the effect of thiols on that measurement. In the case of gGAPDH, the value was approximately one-half of the original after thiol modification, suggesting thiol interference. Nevertheless, gGAPDH may contain a fructosamine structure. beta-Mercaptoethanol itself gave a strong positive result in the fructosamine assay. Hence, thiol groups on glycated proteins should be modified before doing a fructosamine assay because of their substantial interference.

Blood↗

A synthetic lipopolysaccharide-binding peptide based on the neutrophil-derived protein CAP37 prevents endotoxin-induced responses in conscious rats.

The lipid A component of lipopolysaccharide (LPS) derived from Escherichia coli has been implicated as a significant mediator in the development of circulatory and metabolic dysfunction and lethality associated with sepsis. A synthetic peptide corresponding to amino acid residues 20 through 44 of the neutrophil-derived 37-kDa cationic antimicrobial protein (CAP37 P(20-44)) possesses lipid A binding characteristics which may be useful in attenuating in vivo responses induced during circumstances of endotoxemia, including sepsis. The E. coli LPS to be used in the in vivo study was shown to be attenuated by CAP37 P(20-44) in a dose-dependent manner in the in vitro reaction with Limulus amoebocyte lysate. Intravenous infusion of CAP37 P(20-44) (1.5 or 3.0 mg/kg of body weight) with E. coli LPS (250 microg/kg over 30 min) into conscious, unrestrained rats prevented LPS-induced hyperdynamic and hypodynamic circulatory shock, hyperlactacidemia, and leukopenia in a dose-related fashion. CAP37 P(20-44) (0.2, 1.0, and 5.0 mg/kg) administered intravenously to conscious, actinomycin D-sensitized rats following a lethal dose of LPS neutralized LPS toxicity, resulting in dose-dependent 7-day survival rates of 30, 50, and 80%, respectively. CAP37 P(20-44) (5.0 mg/kg) significantly inhibited the endotoxin-induced increase in circulating tumor necrosis factor alpha in sensitized rats. These data demonstrate that CAP37 P(20-44) has the capacity to abolish in vivo biological responses to LPS that are relevant to human sepsis and to significantly neutralize the toxicity of circulating E. coli LPS.

Amino Acid Sequence↗

[Insulin receptors in diabetes mellitus].

We described the clinical significance of the measurement of insulin receptors and the mechanism of non-insulin dependent diabetes mellitus (NIDDM). Changes in insulin receptor (IR) of red blood cells in 47 patients with NIDDM were compared with those of 28 normal controls. The levels of high and low-affinity IR in the 47 patients with NIDDM were lower than those of normal controls and were prominent in untreated new NIDDM patients. Hyperinsulinemia was not found in these patients, indicating that the low IR is a primary defect not caused by the "down regulation" mechanism. In the IDDM patients the levels of high and low-affinity IR were higher than those of the normal controls, but decreased markedly after insulin therapy, indicating that the "up regulation" is attributable to the high IR levels, other than the primary IR defect.

Adolescent↗

[nm23 expression and its correlation with lung metastasis in human salivary adenoid cystic carcinoma].

The nm23 gene products, nucleoside diphosphate kinase (NDPK), expression in salivary adenoid cystic carcinoma (ACC) was evaluted by using LSAB immunohistochemical method. Of 25 cases tested, 16 (64.0%) showed positive staining, in which, higher incidence of positive staining was found in ACC without lung metastasis (88.2%, 15/17) than in that with lung metastasis (12.5%, 1/8; P < 0.01). Expression of NDPK/nm23 was correlated with the P-TNM pattern (P < 0.05), otherwise it was not correlated with pathologic types (P > 0.05). The result suggest that the nm23/NDPK may play a role in suppressing the metastatic potential in ACC.

Carcinoma, Adenoid Cystic↗

[Effects of SAD on cytogenetics of mouse embryonic cells and its embryo development].

We report here that SAD may not only suppress transcription activity of rRNA genes, but also increase SCE frequency of mouse embryonic cells, leading to bloching of the embryo development and fetal death, and finally significant decrease of birth percentage of the new born offsprings. Ata concentration of SAD in 10(-5)mol/L, the transcription activity of rRNA genes was decreased to 15% of the normal level, and the SCE frequency was increased doubly, leading to that the birth percentage was decreased to 50% of normal one. When SAD concentration of 10(-3)mol/L was used the transcription activity of rRNA genes was completely suppressed and SCE frequency was increased to as 4.8 times high as normal, leading to that the birth percentage dropped to 4% from 100%. Our results indicates that both the cytogenetic effects and the inhibition of the embryonic cells are directly proportional to the SAD concentration used. The relationship between the change of both the rRNA gene transcription and SCE frequency and the degree of embryo development are also discussed preliminarily.

Aldehydes↗

The central distribution of adrenomedullin and its effects on blood pressure and heart rate in rats.

The present study was designed to make certain whether there exists adrenomedullin (ADM) in the rat central nervous system and evaluated the hemodynamic actions of intracerebroventricular administration (ICVA) of human ADM[13-52]. By immunohistochemistry (ABC method). We found that there was a discrete localization of ADM-positive immunoreactivity in the rat central system including cerebral cortex, paraventricular tissues, hypothalamus, cerebella cortex, mesencephalon and medulla oblongata. By reverse transcription-polymerase chain reaction (RT-PCR) analysis, rat ADM mRNA was found to be expressed in rat brain. These above results of immunohistochemistry and RT-PCR suggest that ADM exists in the rat brain. We also found that centrally administered ADM[13-52] in a dose of 0.4 to 3.2 nmol/kg provoked marked, prolonged and dose-dependent increases in mean arterial blood pressure (MABP) and heart rate (HR). To clarify the mechanisms of the hemodynamic changes induced by centrally administered ADM [13-52], the effect of centrally administered ADM[13-52] on renal sympathetic nerve activity (RSNA) was studied. The result showed that centrally administered ADM [13-52] (1.6 nmol/kg) provoked a marked increase in RSNA, therefore, the increases in MABP and HR induced by centrally administered ADM [13-52] might be due to the stimulation of central sympathetic mechanism. In addition, we also compared the relationship of activity and structure among the different fragments of ADM. In conclusion, ADM exists in the rat brain, and it may play an important role in the central control of cardiovascular system.

Adrenomedullin↗

[Observation of proliferative capacity of rat salivary gland cells].

Proliferative potentiality of adult rat salivary gland cells was studied by means of bromodeoxyuridine (BrdU) labeling in different time and ABC immunohistochemistry. The labeling rate of intercalated cells in different time after BrdU injection manifest the character of stem cell which is concordant with the theory of semipluripotential bicellular reserve cell suggested by Eversole and Batsakis. This initial labeling rate of intercalated cell is very low and it rises very rapidly to 78 times within 5 days. On the other hand, it also goes down very fast. We proved that the acinar cells and striated cells have the capacity of proliferation which is not so high as intercalated cells for self renewal that can complete the theory of semipluripotential bicellular reserve cell and is beneficial to explain why there is a variety of salivary gland neoplasms.

Animals↗

[The selection of highly lung metastatic salivary adenoid cystic carcinoma clone].

After 5 times repeated selection in vivo, combined with cloning technique in vitro and analysing of platelet aggregation activity, we had selected a highly lung metastatic salivary adenoid cystic carcinoma clone M-5clone24 (Acc-M). Compared with Acc-2, its metastatic rate was 96% vs. 18%; the weight of metastatic lung was 0.88 g vs. 0.31 g. The metastatic rate and the weight of metastatic lung positively correlated with platelet aggregation activity. The aggregation activity might be used as a useful parameter to assess Acc metastasis potential.

Animals↗