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M Li

Publications and source records attributed to M Li.

At least 1,027 records · Page 57Linked to original sources

Functional modulation of brain sodium channels by cAMP-dependent phosphorylation.

Voltage-gated Na+ channels, which are responsible for the generation of action potentials in brain, are phosphorylated by cAMP-dependent protein kinase in vitro and in intact neurons. Phosphorylation by cAMP-dependent protein kinase reduces peak Na+ currents 40%--50% in membrane patches excised from rat brain neurons or from CHO cells expressing type IIA Na+ channels. Inhibition of basal cAMP-dependent protein kinase activity by transfection with a plasmid encoding a dominant negative mutant regulatory subunit increases Na+ channel number and activity, indicating that even the basal level of kinase activity is sufficient to reduce Na+ channel activity significantly. Na+ currents in membrane patches from kinase-deficient cells were reduced up to 80% by phosphorylation by cAMP-dependent protein kinase. These effects could be blocked by a specific peptide inhibitor of cAMP-dependent protein kinase and reversed by phosphoprotein phosphatases. Convergent modulation of brain Na+ channels by neurotransmitters acting through the cAMP and protein kinase C signaling pathways may result in associative regulation of electrical activity by different synaptic inputs.

Animals↗

Comparison of the expression of a mutant dihydrofolate reductase under control of different internal promoters in retroviral vectors.

To determine the effect of different promoters on the expression of an altered dihydrofolate reductase (DHFR) gene conferring methotrexate (MTX) resistance in different cell types, double-copy retroviral vectors were constructed carrying a murine mutant DHFR under the control of five different promoters, i.e., human adenosine deaminase (ADA), simian virus 40 (SV40), thymidine kinase (TK), human beta-actin, and cytomegalovirus (CMV). Their expression was compared in NIH-3T3 cells, three human leukemia cell lines, and mouse bone marrow. The variant DHFR is readily expressed from these various promoters in retroviral vectors at a selectable level. In 3T3 cells, the DHFR constructs containing the SV40 promoter conferred the highest levels of resistance to MTX. In K562 and Raji cells, the construct with the TK promoter produced the highest level of resistance. However granulocyte-macrophage colony-forming unit (CFU-GM) colonies from mouse marrow were more resistant to MTX when infected with vectors containing the SV40 promoter and ADA promoter as compared to the other promoter constructs. These studies show that mouse fibroblast cell lines such as NIH-3T3 do not predict the effectiveness of retroviral-mediated gene transfer for marrow progenitor cells, and that the activity of retroviral vector-encoded promoters vary in an unpredictable manner from cell type to cell type. Possible implications for basic gene transfer studies and clinical applications are discussed.

3T3 Cells↗

Role of the renal medulla in adrenocorticotrophin-induced hypertension in rats.

OBJECTIVES: The mechanism of hypertension induced by adrenocorticotrophin (ACTH) remains unclear. The antihypertensive renomedullary lipids are vasodilators and it has been proposed that a deficiency of these lipids may contribute to the hypertension produced by destruction of the renal papilla. The aim of the present work was to study ACTH hypertension in both control and chemically renomedullectomized rats. METHODS: Renomedullectomy was produced by single intraperitoneal injection of 2-bromoethylamine (BEA) at 400 mg/kg. RESULTS: BEA-treated rats all developed increases in water intake and urine volume, with loss of papillae and medullary and cortical fibrosis. There was a significant correlation between papillary ablation and systolic blood pressure (SBP). SBP in renomedullectomized rats was higher after ACTH than sham injection, and higher than after ACTH injection in intact rats. CONCLUSION: Chemical renomedullectomy with BEA did not block or attenuate the onset or magnitude of ACTH hypertension in the rat.

Adrenocorticotropic Hormone↗

Inhibition of NO synthesis has an additive effect on hypertension induced by ACTH in conscious rats.

1. The haemodynamic and metabolic effects of oral intake of approximately 30 mg/kg per day N-nitro-L-arginine (NOLA) were examined in sham and adrenocorticotrophin (ACTH, 0.5 mg/kg per day) treated conscious Sprague-Dawley rats (n = 33). 2. NOLA administration produced an increase in systolic blood pressure of 24 +/- 6 mmHg (P < 0.001), but did not alter food or water intake, urine volume or electrolyte excretion in rats not treated with ACTH. 3. Compared with sham injection, ACTH-treated rats demonstrated an increase in systolic blood pressure (water + sham, 3 +/- 1 mmHg; water + ACTH, 16 +/- 3 mmHg; P < 0.001), loss of bodyweight, and increases in water intake and urine volume. 4. The magnitude of the blood pressure rise in ACTH-treated rats was greater in those receiving NOLA than in those drinking water only (water + ACTH, 16 +/- 3 mmHg; NOLA + ACTH, 37 +/- 3 mmHg; P < 0.05). Metabolic changes were similar. 5. Inhibition of nitric oxide is unlikely to be a major determinant of ACTH-induced hypertension in the rat, since NOLA increased blood pressure whether or not ACTH was administered, indicating an additive effect of ACTH and NOLA administration.

Adrenocorticotropic Hormone↗

Cloning and characterization of the groESL operon from Bacillus subtilis.

The sequence of the 10 N-terminal amino acids of a Bacillus subtilis protein that cross-reacts with antibody to Escherichia coli GroEL was used to design a set of degenerate oligonucleotide probes. These probes identified a clone which carries almost the entire groESL operon from a B. subtilis subgenomic library. By chromosomal walking, an additional fragment carrying the 3' end of groESL and its flanking sequence was isolated. Sequence analysis revealed two open reading frames (ORFs) in the cloned DNA. The upstream ORF encodes a 10-kDa protein which has 47% amino acid identity with E. coli GroES. The downstream ORF encodes a 58-kDa protein which is 62% identical to E. coli GroEL. A 2.1-kb groESL mRNA from B. subtilis was detected independently by Northern (RNA) blot analyses with a groES- and a groEL-specific probe. This demonstrated that groES and groEL are in an operon. The groESL promoter was located by using a promoter-probing plasmid, and the apparent transcription start site was mapped by primer extension analysis. The same promoter is utilized under normal and heat shock conditions. This promoter has the same features as a typical sigma A promoter. A strain in which the groESL operon was under the control of the sucrose-inducible sacB promoter was created. With this strain, it was possible to show that both groES and groEL are essential genes under both normal and heat shock conditions.

Amino Acid Sequence↗

Effects of methylguanidine on sodium and organic ion transport.

We studied the acute effect of methylguanidine (MG), a suspected uremic toxin that accumulates in renal failure, on p-aminohippurate (PAH) and tetraethylammonium (TEA) uptake in rabbit kidney slices, on Na+,K+ ATPase activity in the microsomal fraction of rabbit kidneys, and on transepithelial active Na transport across toad skin. MG at concentrations ranging from 0.05 (similar to that reported in uremic patients) to 1.0 mM does not affect the organic anion (PAH) uptake, although it exhibits a concentration-dependent inhibition of organic cation (TEA) uptake. MG at concentrations from 0.05 to 5 mM had no effect on kidney Na+,K+ ATPase activity or on active transepithelial Na transport across toad skins when applied to the outside bathing solution; however, MG (greater than 1 mM) stimulated Na transport when applied to the inside bathing solution. These results are not consistent with the hypothesis that MG is a potential uremic toxin that causes the natriuresis and other toxic effects. However, long-term toxic effects of MG on the kidney were not assessed in the present study.

Analysis of Variance↗

Muscarinic cholinergic stimulation elevates intracellular pH in chicken granulosa cells by a Ca(2+)-dependent, Na(+)-independent mechanism.

Various hormones and growth factors act at least in part by raising the cytosolic pH of the target cell. In the present studies we examined the influence of the muscarinic cholinergic agonist carbachol on intracellular pH (pHi) in chicken granulosa cells. The pHi in granulosa cells from the two largest preovulatory follicles of hens was determined spectrofluorometrically using the pH-sensitive dye 2',7'-bis-(carboxyethyl)- 5(6)-carboxyfluorescein (BCECF). Carbachol (0.1-2 mM) induced a concentration-dependent increase in pHi, which reached a maximum of 0.23 +/- 0.02 pH units at a concentration of 1 mM. Cytosolic alkalinization was observed within 10 min of the addition of carbachol and lasted over the 60-min observation period. The effect of carbachol was mimicked by acetylcholine (1 mM) and muscarine (1 mM), but not by nicotine (0.1 mM). The carbachol-induced alkalinization was blocked by pretreating the cells with the muscarinic cholinergic receptor antagonists atropine (1 and 10 microM) and pirenzepine (0.01 and 0.1 mM). The increase in pHi did not appear to be mediated by a conventional mechanism involving the Na+/H+ antiporter, because it was unaffected by replacement of extracellular Na+ with the nonpermeant choline chloride or the presence of antiporter inhibitors, such as amiloride, dimethylamiloride, or ethylisopropylamiloride. However, the mechanism required external Ca2+, but did not involve Ca2+ channels, because it was unaffected by the general Ca2+ channel blocker methoxyverapamil (50 microM). Thus, Ca(2+)-dependent and Na(+)-independent intracellular alkalinization may be a part of the signalling mechanism by which muscarinic receptor activation regulates chicken granulosa cell function.

Animals↗

Calcium ionophores increase intracellular pH in chicken granulosa cells.

Several hormone agonists exert their physiological actions by triggering an inositol phospholipid-Ca2+ signalling cascade and cytosolic alkalinization. Although calcium ionophores have been used extensively to probe the role of Ca2+ in the regulation of steroidogenesis in granulosa cells, the precise relationship between changes in intracellular Ca2+ (Ca2+i) and pH (pHi) is unclear. In the present study we have used a fluorescent pH indicator, 2'7'-bis-(2-carboxyethyl)-5(and-6)-carboxyfluorescein, to examine the influence of two Ca2+ ionophores, ionomycin and 4-Bromo-A23187 (4-Br-A23187), on pHi in chicken granulosa cells. Chicken granulosa cells from the largest preovulatory follicle were incubated with Ca2+ ionophores (0-2 microM) and/or inhibitors of Na+/H+ antiport (amiloride, dimethylamiloride and ethylisopropyl amiloride; 0.5, 5 and 50 microM respectively) in the presence of Na+ (or choline+; 0-144 mM) and/or Ca2+ (0-10 mM). Ionomycin or 4-Br-A23187 elicited a rapid and sustained cytosolic alkalinization. The magnitude of increase in pHi was dependent on the concentration of the Ca2+ ionophore and the presence of extracellular Ca2+ but independent of extracellular Na+. Pretreatment of the cells with amiloride or its analogues failed to affect the increase in pHi induced by the Ca2+ ionophores significantly. These findings demonstrate that, in addition to their widely reported effects on Ca2+i redistribution in granulosa cells, 4-Br-A23187 and ionomycin cause Ca(2+)-dependent cytosolic alkalinization. This action of the Ca2+ ionophores is independent of the Na+/H+ antiport. Caution must be exercised in using Ca2+ ionophores as probes to define the role of Ca2+ in the regulation of granulosa cell function.

Amiloride↗

ACTH hypertension in the rat: role of sodium chloride.

Adrenocorticotrophin (ACTH) produces adrenally dependent increases in both blood pressure and salt (NaCl) appetite in the rat. The present study examines the effect of free access to a high intake of NaCl on ACTH hypertension in the rat to test the hypothesis that high NaCl intake would amplify the rise in blood pressure. Either water or 1%NaCl were offered to sham or ACTH treated Sprague-Dawley rats. Subcutaneous injections of synthetic ACTH (0.5 mg/kg/day) for 10 days caused large increases in the intake of both 1%NaCl (+240 +/- 6 ml/day) and water (+45 +/- 4 ml/day), urine volume (1%NaCl + ACTH +182 +/- 4 ml/day, ACTH + water +36 +/- 2 ml/day), adrenal weight (ACTH + water 176 +/- 18, ACTH + 1%NaCl 367 +/- 129 mg/100 g body weight) and maximum systolic blood pressure (SBP) (ACTH + water +18 +/- 5 mmHg; 1%NaCl + ACTH +16 +/- 3 mmHg). Some ACTH + 1%NaCl rats developed severe edema. Thus, free access to NaCl in ACTH treated Sprague Dawley rats did not potentiate the hypertension, but was associated with edema in some animals.

Adrenal Glands↗

Screening of bacterial strains producing maltotetraose-forming amylase and the conditions for enzyme production.

The authors isolated 1380 bacterial strains from 290 soil samples collected in China and 490 strains were received from other research teams in this institute. By screening 707 strains showed starch-hydrolyzing activity. By further screening and paper chromatographic test, three strains with maltotetraose as the major product were obtained. The maltotetraose was further confirmed by treatment with beta-amylase splitting to maltose and with glucoamylase to glucose. The most promising strain was 537.1, which produced maltotetraose about 90% (w/w) in the starch hydrolysate. While the other two strains produced maltose and maltotriose besides maltotetraose. Strain 537.1 was tentatively identified as Alcaligenes sp. The optimum conditions for enzyme production were as follows: medium composition: 1.5% maltose; 0.5% peptone with initial pH of 7.0-7.5; cultured at 27-28 degrees C for 48 hours on rotary shaker. The culture supernatant of the strain 537.1 can hydrolyze starch and different kinds of cereal flour with a high yield of maltotetraose in the hydrolysate.

Bacteria↗

[Detection of anti-idiotypic antibody specific for anti-HBs in HBsAg positive serum].

An ELISA for detecting anti-idiotypic antibody specific for anti-HBs(anti-anti-HBs, Ab 2) was devised and used to detect twenty-three asymptomatic HBsAg positive sera. Of these HBsAg positive samples, five were positive for anti-anti-HBs. Three anti-anti-HBs positive samples and two anti-anti-HBs negative samples were checked by immune electron microscopy (IEM). Non HBV particles were found in the anti-anti-HBs positive samples, but the particles were seen in one of the anti-anti-HBs negative samples. According to the immune networks theory, the anti-anti-HBs with the characteristic of HBsAg internal can mimic HBsAg and bind to anti-HBs(Ab 1), so that the anti-anti-HBs may interfere the routine detection of HBsAg and results in false positive. The data described above demonstrated the existence of this consideration. The authors suggested that samples with HBsAg positive, especially those from asymptomatic carriers should be detected for anti-anti-HBs in order to rule out the interference of anti-anti-HBs.

Antibodies, Anti-Idiotypic↗

[Electrical stimulation in the treatment of osteoporosis in sciatic denervated rat tibia].

Osteoporosis in the sciatic-denervated rat tibia was reversed with a capacitively coupled electrical field. In the reversal of a previously established osteoporosis, a statistically significant enhancement of wet weight, dry weight, ashed weight, ultimate strength, cortical area, cortical thickness and a concomitant decrease in cortical porosity occurred in the stimulated, denervated tibiae of the experimental animals compared with the nonstimulated, denervated tibiae of the control animals. A 60 Hz symmetrical sinewave signal was effective in reversing osteoporosis at 10 V peak to peak. Reversal of a well-established osteoporosis in laboratory animals has not been reported previously.

Animals↗

Differential regulation of subtypes m1-m5 of muscarinic receptors in forebrain by chronic atropine administration.

The regulation of individual muscarinic receptor subtypes in rat cerebral cortex/dorsal hippocampus was examined following 14-day administration of the nonselective antagonist atropine. Total muscarinic receptor density increased 24%, from 2196 fmol/mg to 2722 fmol/mg. The nature of this increase was examined using a panel of antisera selective for the m1 to m5 muscarinic receptors. Thus, 97% of all cortical/hippocampal receptors were accounted for by immunoprecipitation. Three subtypes were observed to increase significantly in density: m1 receptor from 824 to 982 fmol/mg (19%, P less than .05); m2 receptor from 476 to 519 fmol/mg (9%, N.S.); m3 receptor from 259 to 438 fmol/mg (69%, P less than .001); m4 receptor from 574 to 638 fmol/mg (11%, P less than .05); and the m5 receptor from 23 to 38 fmol/mg (65%, N.S.). Receptors coupled to the hydrolysis of phosphoinositides (m1, m3, m5) appeared to be preferentially up-regulated (32% over control levels, P less than .001) compared with those coupled to the inhibition of adenylyl cyclase (m2, m4; 10% over control levels, P less than .05). The absolute density of the molecularly defined m1 and m4 receptors, which reportedly possess the highest affinity for pirenzepine (M1), increased 16% (P less than .05), whereas the density of receptors having the lowest affinity for pirenzepine (m2, m3 and m5) increased 31% (P less than .001) with atropine treatment. When the increase in total receptor density was examined with [3H]pirenzepine, the 16% elevation in high-affinity (m1 and m4) sites was not detected.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Regularity of recurrent sites of cervix carcinoma after radical operation].

Radical operations were performed in 872 cases of cervical carcinoma from April 1964 to December 1985. The concrete ways of operation and auxiliary therapy were different according to clinical stage of each case. All the cases were followed-up for more than 5 years. 114 cases showed definite single or multiple lesions. Analysis of 92 cases of single lesions, the revealed that recurrent sites may be related to clinical stage, pathology, pelvic node metastases and auxiliary treatment. The local recurrence rate of single lesions was 59.8% and that of distant lesions was 40.2%.

Female↗

[Gene deletion of X-linked ichthyosis].

The characterization of steroid sulfatase (STS) gene mutation from seven X-linked ichthyotic patients was performed by multiple polymerase chain reaction (MPCR) which amplified two specific regions at the 5' and 3' end of STS gene. The results indicated that five out seven patients were found to have entire STS gene deletion. Two other cases and five patients' mothers showed two amplification fragments. So did two cases of dominant ichthyosis vulgaris. It was further ascertained that entire gene deletion is the main mutation of STS locus in Chinese population. MPCR is a rapid and simple method for gene diagnosis of X-linked ichthyosis.

Arylsulfatases↗