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Biomedical subjects

H Mao

Publications and source records attributed to H Mao.

64 records · Page 4Linked to original sources

[Determination of urea in milk by enzyme coupled electrode method].

An Enzyme coupled electrode method for the determination of urea content in milk was studied systematically. The linear range of the method, the detection limit, recovery and relative standard deviation are 2.1-2.1 x 10(3) micrograms/ml, 2 micrograms/ml, 95.3%-101.0% and 1.5%-3.9%, respectively. The reliability of the method was confirmed by comparison with colorimetry (0.2 < P < 0.05). Using this method, we determined the content of urea in seven milk samples from Chengdu. The values range from 152.2-288.5 micrograms/ml and agree with those reported by other authors. This method is simple, rapid, accurate and inexpensive.

Animals↗

Depolarization of cell membrane is associated with an increase in ciliary beat frequency (CBF).

We hypothesize that activation of muscarinic cholinergic receptors depolarizes the cell membrane of the mammalian ciliated cells which in turn causes an increase of CBF. To test this hypothesis, a di-8-ANEPPS fluorescence photon counting and nonstationary heterodyne laser light scattering system was developed to measure cell membrane potential (psi) and CBF in cultured ovine tracheal ciliated cells simultaneously. Carbachol dose dependently depolarized the cell membrane with a corresponding stimulation of CBF. The carbachol induced depolarization of cell membrane and increases of CBF were inhibited by prior application of either atropine or verapamil or amiloride. These novel data suggest that depolarization of the cell membrane and the corresponding stimulation of CBF caused by the activation of muscarinic receptors of the mammalian ciliated cells are dependent on the influx of either extracellular Ca2+ or Na+.

Amiloride↗

Some properties of the ecdysteroid receptor in the salivary gland of the ixodid tick, Amblyomma hebraeum.

Salivary gland degeneration in ixodid ticks is triggered by an ecdysteroid hormone. We used [3H]ponasterone A (PoA) as a specific ligand to detect the ecdysteroid receptor in the salivary glands of large, partially fed female ticks (Amblyomma hebraeum Koch; Acari: Ixodidae). Binding of [3H]PoA was thermolabile and sensitive to pronase, but not to DNase or RNase, indicating that the ligand binds to a protein. Scatchard analysis of [3H]PoA binding strongly suggested the presence of an ecdysteroid receptor in cytosolic and nuclear extracts of the tissue. The Kd and Bmax for PoA binding in cytosol were 0.72 +/- 0.09 nM and 175 +/- 12 fmol/mg protein, respectively (n = 8). Corresponding figures for nuclear extract were 1.1 +/- 0.5 nM and 282 +/- 35 fmol/mg protein, respectively (n = 3; P > 0.05 compared to cytosol). The relative ability of unlabeled ecdysteroids to compete for [3H]PoA binding was (in descending order): PoA > muristerone A > makisterone A > 20-hydroxyecdysone > mesylinokosterone > ecdysone. The Kd estimated for 20-hydroxyecdysone (probably the natural hormone) correlates very well with its physiological potency in inducing salivary gland degeneration in vivo and in organ culture. None of the vertebrate steroids tested (estradiol, testosterone, progesterone, and corticosterone) was able to displace PoA binding at a concentration 10(5) times higher than PoA. The cytosolic form of the receptor migrated to the 3.2 S region of a 10-40% sucrose density gradient.

Animals↗

Interaction of HIV Rev peptides with the Rev response element RNA.

As a model system for understanding RNA-protein interactions, we are studying the structure of peptides derived from HIV Rev in complex with an RNA derived from the Rev Response Element (RRE) using NMR spectroscopy. Formation of the Rev peptide-RRE complex is accompanied by formation of two purine-purine base pairs in the RRE. These base pairs create an unusual geometry that widens the major groove of the RRE RNA. The Rev peptide is predominantly in an alpha-helical conformation in the complex. The specific recognition of the RRE by Rev involves widening of the major groove to accommodate the Rev alpha-helix.

Base Sequence↗

Synergistic antitumor effect of tumor necrosis factor-related apoptosis-inducing ligand combined with cisplatin in ovarian carcinoma cell lines in vitro and in vivo.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been shown to exert selectively cytotoxic activity against many tumor cells but not normal cells. In this study, we evaluated the antitumor activity of TRAIL and cisplatin (CDDP) both separately and combined in the human ovarian cancer cell lines. In vitro study showed that TRAIL elicited significant cell apoptosis of cell lines 3AO, SKOV3, and OVCAR3 in a dose- and time-dependent manner (P < 0.05), while normal ovarian epithelial cells were resistant; this toxicity-free effect may be the result of upregulation of TRAIL receptors DcR1 and DcR2. Combined TRAIL and CDDP therapy produced more profound cell killing in 3AO cells than each alone (P < 0.05), and CDDP could upregulate the expression of both death and decoy TRAIL receptors. To further evaluate the apoptosis-inducing effects of TRAIL and the combination therapy, the abdominally and subcutaneously spread tumors in nude mice via inoculation of 3AO cells were established, and treatment of TRAIL resulted in a dose- and time-dependent inhibition of tumor growth while slight damage was observed in normal tissues. Furthermore, combined TRAIL and CDDP therapy had a synergistic effect in the regression of established ovarian cancer xenografts than TRAIL treatment alone (P < 0.05). We also examined the apoptosis-related gene expression in the transplantation tumors after TRAIL treatment, and the data suggested that the intracellular mechanism of TRAIL may be associated with downregulation of Bcl-2 and upregulation of CD95 and Apo2.7.

Adenocarcinoma, Mucinous↗

Fluorescence and laser photon counting: measurements of epithelial [Ca2+]i or [Na+]i with ciliary beat frequency.

We describe a system we developed that enabled simultaneous measurements of either epithelial calcium ion concentration ([Ca2+]i) or sodium ion concentration ([Na+]i) with the ciliary beat frequency (CBF) in native ciliated epithelia using either Fura-2 (AM) or SBFI (AM) ratiometric fluorescence photon counting along with nonstationary laser light scattering. Studies were performed using native epithelial tissues obtained from ovine tracheae. The dynamic range of the laser light-scattering system was determined by a simulated light "beating" experiment. The nonstationary CBF was demonstrated by the time-frequency analysis of the raw photon count sequences of backscattered heterodyne photons from cultured and native epithelia. Calibrations of calcium and sodium ion concentrations were performed using the respective Fura-2 and SBFI impermanent salts as well as in native epithelia. The cumulative responses of 10(-6), 10(-5), and 10(-4) M nifedipine on [Ca2+]i together with the CBF as well as the cumulative responses of 10(-5), 10(-4), and 10(-3) M amiloride on [Na+]i together with the CBF were also determined. Nifedipine decreased [Ca2+]i but had no effect on CBF. Amiloride decreased [Na+]i and CBF. Stimulation of CBF corresponded with either an increase of [Na+]i or an increase of [Ca2+]i. Decreases of [Na+]i or substantial decreases of [Ca2+]i were associated with decreases in the CBF. These data demonstrate the utility of this system for investigating the regulatory mechanisms of intracellular ions dynamics and the CBF in native epithelia.

Amiloride↗