Creutzfeldt-Jakob disease in a Chinese patient with a novel seven extra-repeat insertion in PRNP.
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Adult↗
Biomedical subjects
D X Wang
Publications and source records attributed to D X Wang.
Herpes simplex virus 1 inhibits apoptosis through a caspase-3-dependent pathway in primary cultures of cortical neuronal cells of fetal mice.
We could induce apoptosis in primary cultures of cortical neurons of fetal mice with ceramide or sorbitol. The induction was accompanied by an increase in caspase-3 (CAS-3) activity and depolarization of the inner mitochondrial membrane of neuronal cells which both could be reversed by Herpes simplex virus 1 (HSV-1) infection. We conclude tha HSV-1 infection inhibited the apoptosis, induced in neuronal cells by sorbitol or ceramide, via a CAS-dependent pathway.
Animals↗
Telomere erosion is independent of microsatellite instability but related to loss of heterozygosity in gastric cancer.
AIM: To correlate the length of the telomere to microsatellite instability (MSI) and loss of heterozygosity (LOH) of APC, MCC and DCC genes in gastric carcinomas. METHODS: Telomeric restriction fragment (TRF) length of gastric cancer was measured with Southern blot. LOH of APC, MCC and DCC genes, microsatellite instability (MSI) and frameshift mutation of hMSH6, TGF-betaRII and BAX genes were analyzed by PCR-based methods. RESULTS: Sixty-eight cases of sporadic gastric carcinoma were studied for MSI using five microsatellite markers. MSI in at least one locus was detected in 17 (25%) of 68 tumors analyzed. Frameshift mutations of hMSH6, TGF-betaRII and BAX were detected in 2,6 and 3 of gastric carcinomas respectively showing high MSI (> or = 2 loci, n = 8), but none was found in those showing low MSI (only one locus, n = 9) or MSS (tumor lacking MSI or stable, n = 51). Thirty-five cases, including all high MSI and low MSI, were studied for TRF. The mean TRF length was not correlated with clinicopathological parameters. No association was observed between TRF length and MSI or frameshift mutation. On the contrary, LOH at the DCC locus was related to telomere shortening (P<0.01). This tendency was also observed in APC and MCC genes, although there was no statistical significance. CONCLUSION: The development of gastric cancer can arise through two different genetic pathways. In high MSI gastric cancers, defective mismatch repair allows mutations to accumulate and generate the high MSI phenotype. In gastric cancers showing either low MSI or MSS, multiple deletions may represent the LOH pathway. Telomere erosion is independent of high MSI phenotype but related to the LOH pathway in gastric cancer.
Adult↗
[Determination of the pKa value of icariin and its content in the Chinese herb medicine epimedium grandiflorum morr. by capillary electrophoresis].
Knowledge of dissociation constants is important for prediction and understanding the migration behavior of analytes in capillary electrophoresis. Icariin is the active component of the Chinese herb medicine Epimedium grandiflorum Morr.. In order to determine the dissociation constant of icariin and to show many important pharmacology activities, a CZE method has been used to determine the ion mobility (mu A-) and the pKa value of icariin based on the non-linear relation between mu eff and [H+] and the linear relation between the reciprocal of effective mobility of the solute (1/mu eff) and the [H+] of the buffer solution. In addition, the change of pKa of icariin with the increase of ethanol concentration in the buffer was also investigated. Under the buffer condition of 24 mmol/L phosphate + 30% ethanol, the content of the active component icariin in Chinese herb Epimedium grandiflorum Morr. was quantitatively determined. The linear correlation equation was Y = 6.96 x 10(-3) + 17.0X, and the linear range is 0.032 g/L-0.354 g/L.
Electrophoresis, Capillary↗
[Studies on the synthesis and bioactivity of alpha-alkylaminobenzyl-phosphonic acids].
AIM: To search for some substituted benzyl phosphonic acids as leading compounds with inhibiting effect on osteoclast formation. METHODS: Target compounds were prepared from aromatic aldehydes, primary amine and phosphorous acid using tetramethylenesulfone as solvent via Arbuzov type reaction. The effect on inhibiting the formation of osteoclast-like cells (OLC) of related compounds was studied by incubating the extract of rat femur marrow. RESULTS: Ten compounds of alpha-alkylaminobenzyl phosphonic acids have been synthesized and identified by MS or 1HNMR analysis. Three (2, 8 and 9) of them were found to have notable effect on the inhibition of OLC formation (P < 0.01). CONCLUSION: Among the present substituted benzyl phosphonic acids, the increased aromaticity and hydrophobicity (such as compound 9) can remarkably enhance the ability to inhibit OLC formation.
Animals↗
Prolactin concurrently activates src-PLD and JAK/Stat signaling pathways to induce proliferation while promoting differentiation in embryonic astrocytes.
In normal development, embryonic astrocytes progress through their cell lineage by acquiring differentiation, by apoptosis, and by proliferation. In this study, we show that embryonic astrocytes may maintain and make gains in differentiation as they simultaneously progress through one cell cycle when induced by prolactin (PRL). Prolactin induced the majority of astrocytes to incorporate bromodeoxyuridine (BrdU) with a four-fold increase over controls after 18 h of exposure. Investigating possible mitogenic signaling pathways we show for the first time that prolactin is coupled to a sustained phospholipase D (PLD) activation, with an efficacy similar to the phorbol ester and astrocytic mitogen 12-tetradecanoylphorbol-13-acetate (TPA). Both cyclosporine and suramin abolished this activation. Staurosporine and calphostin C also inhibited the PRL effect by 50%, consistent with involvement of protein kinase C-(PKC)-alpha, the major PKC isoform in astrocytes. Genistein and PP1 blocked the activation indicating additional regulation by cytosolic tyrosine kinases. This profile of PLD activation was suggestive of a PLD I isoform and a mitogenic response. Upon completion of the cell cycle, analysis of glia fibrillary acidic protein (GFAP) and vimentin abundance, and glutamine synthetase (GS) activity showed that astrocytes had gained in expression of differentiation markers. Moreover, the intensity of GFAP immunofluorescence was greater per cell, as was the length of the cell processes. In exploring the signaling for prolactin-induced differentiation we found that prolactin activated the tyrosine kinase Janus kinase (JAK) 2 and significantly stimulated tyrosine, phosphorylation of the prolactin receptor. Stat 1 and 3 were also activated presumably downstream to JAK2 activation. A rapid translocation of the cytosolic Stats over the nucleus was seen in nearly every astrocyte corresponding well with the gains in GFAP per cell. The Stats translocation did not depend on MEK-ERK inhibition by PD98059, inhibition of p38 by 1 microm SB203580, or Src kinase family inhibition by PP1. Our results demonstrate the ability of PRL to concurrently induce activation of PLD, a mitogenic signaling pathway in astrocytes, and prolonged stimulation of Stat1, compatible with the increased GFAP upregulation and cell differentiation. Considered together this data may provide an explanation on the fast gain in both numbers and differentiation in the astrocytic population during development (HD 09402, CRF).
Animals↗
[Mechanisms of the suppression of hypoxic pulmonary vasoconstriction by chronic hypoxia].
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Animals↗
[Determination of the active components in Chinese herb Aloe vera L. var. chinensis (Haw.) Berger by capillary zone electrophoresis].
It has been proved that the Chinese herb Aloe vera L. var. chinensis (Haw.) Berger as well as its active components showed many important pharmacology activities. In order to find an easy and low-cost method to control the quality of the herb, a CZE method for the determination of the active components aloin and aloe-emodin in Aloe vera L. var. chinensis (Haw.) Berger was developed in this work. Under the buffer conditions of 24 mmol/L phosphate (pH 10.52), applied voltage of 15 kV and detector wavelength of 254 nm, baseline separation of the active compounds in Aloe vera L. var. chinensis (Haw.) Berger was achieved and the active components were quantitatively analyzed. The linear calibration equations of the two components are: Y= -0.140 + 57.2X (r = 0.997) for aloin and Y = -0.393 + 1.08 x 10(2) X (r = 0.999) for aloe-emodin respectively. In addition, the effects of buffer pH value and organic modifier on the migration behavior of the solutes were also investigated.
Aloe↗
Infrequent loss of heterozygosity of APC/MCC and DCC genes in gastric cancer showing DNA microsatellite instability.
AIM: To investigate the role of DNA microsatellite instability (MSI) in gastric carcinogenesis by studying associations between MSI status, clinicopathological features, and loss of genetic loci. METHODS: Six microsatellite loci and loss of heterozygosity at APC, DCC, and MCC were analysed by polymerase chain reaction based methods in 53 cases of advanced gastric cancer. RESULTS: MSI was observed in 32.1% of gastric carcinomas (17/53) and 20% of foci of intestinal metaplasia (3/15). Seven gastric carcinomas (13.7%) were MSI-high (MSI-H) (three loci or more) and 10 (18.9%) were MSI-low (MSI-L) (one or two loci). The frequency of MSI-H was higher in intestinal (25.0%) than in diffuse carcinomas (3.7%) (p < 0.05). None of the MSI-H tumours showed loss of heterozygosity at APC, MCC, or DCC loci. CONCLUSIONS: MSI may have an important and early role in a subset of gastric cancers, particularly the intestinal type. The MSI-H subset of gastric cancer has features in common with its colorectal counterpart, whereas MSI-L and microsatellite stable cancers appear to develop through the loss of heterozygosity pathway.
Adult↗
[Effect of ET-1 antisense oligodeoxynucleotide on the hemodynamics of normal and experimental hypertensive rats].
It was previously found that a phosphorothioated antisense oligodeoxynucleotide (ETASODN) significantly inhibits production of endothelin-1 (ET-1). The purpose of the present study was to determine whether intracerebroventricular injection of ETASODN targeted to prepro-ET-1 is capable of exerting the same preventing effect on the aorta narrowing of experimentally modeled hypertensive rats. Radioimmunoassay showed that ET-1 level in the brain stem of hypertensive rats was significantly elevated. In addition to down-regulating the ET-1 level, astisense could also reduce mean arterial pressure (MAP), heart rate and LVSP in model rats. The antisense also down-regulated the ET-1 level in hypothalamus and brain stem, reducing MAP in normal control rats. After treatment with the antisense, the value of delta MAP was markedly lowered in experimental hypertensive rats as compared to the control ones. Thus it appears that (1) ET-1 might play an important role in central cardiovascular regulation in rats and (2) antisense ETASODN might be used in treatment of hypertension via inhibiting ET-1 production.
Animals↗
Autonomous epicardial and endocardial boundary detection in echocardiographic short-axis images.
An autonomous endocardial and epicardial boundary detection (ABD) method is reported. One hundred ten cycles from 55 clinical studies were selected retrospectively. Image sequences were digitized at 512 x 480 pixel resolution. The point-by-point boundary positions of the ABD and the areas enclosed were compared with positions and enclosed areas drawn by three independent observers. Correlation coefficients for epicardial end-diastolic (ED) and end-systolic (ES) areas, endocardial ED and ES areas, muscle area, and fractional area change were 0.970, 0.976, 0.951, 0.985, 0.887, and 0.878, respectively. Bland-Altman analysis showed negligible biases with standard deviations comparable to those of the observers. The mean difference between the ABD border and the consensus observer border positions in 64 directions falls within the mean range of interobserver border positions, suggesting that shape is also well defined by the ABD.
Echocardiography↗
Loss of heterozygosity and loss of expression of the DCC gene in gastric cancer.
AIM: To investigate the role of DCC gene inactivation in the development and progression of gastric cancer. METHODS: Loss of heterozygosity and loss of expression of the DCC gene was studied in 51 surgical specimens of gastric cancer using detection based on polymerase chain reaction. RESULTS: Loss of heterozygosity was found in 35.3% (18 of 51) of specimens and was detected more often in stage III and IV (50%) than in stage I and II cancers (14.3%) (p < 0.05). Occurrence of loss of heterozygosity was not correlated with histological type, tumour size, depth of invasion, or lymph node metastasis. Loss of expression was found in 49% of cases (25 of 51). Loss of expression was not significantly correlated with any clinicopathological variable. CONCLUSIONS: Loss of heterozygosity and loss of expression of the DCC gene are often encountered in gastric cancer. Loss of heterozygosity of the DCC gene is a late event and associated with malignant progression.
Gene Expression↗
[3H]benzylpempidine, a new radioligand for probing a putative channel site on nicotinic cholinergic receptors.
A study was undertaken to assess the receptor binding characteristics of [3H]4-benzylpempidine to an allosteric site on calf brain membranes associated with nicotinic cholinergic receptors and to compare the binding affinity of novel arylpempidine analogs with their ability to antagonize the behavioral effects of nicotine in mice. Scatchard analysis of the binding yielded a K(d) of 20 nM and a B(max) of 330 fmols/mg membrane protein. [3H]4-benzylpempidine appears to be a more satisfactory ligand than [3H]mecamylamine, since it possessed a 50-fold greater affinity and its binding was far less sensitive to inorganic ions and Tris. Among the arylpempidine analogs 4-m-chlorobenzylidenepempidine and 4-benzylidenepempidine had the lowest K(i) values (1.4 nM and 5.0 nM, respectively) and were the most potent in antagonizing nicotine-induced seizures in mice. Although the K(i) values for pempidine and mecamylamine were 1-2 orders of magnitude greater than any of the arylpempidines, the dose required to antagonize nicotine-induced seizures in mice was comparable to the arylpempidines. One explanation for this apparent discrepancy in the correlation of binding affinity and nicotine antagonism is the lower brain penetration of arylpempidines compared to mecamylamine, following their systemic administration to mice.
Animals↗
Expression and characterization of the rat alpha 4 beta 2 neuronal nicotinic cholinergic receptor in baculovirus-infected insect cells.
Baculovirus expression systems have been developed to generate 1) a neuronal nicotinic cholinergic receptor comprising both the alpha 4 and beta 2 subunits and 2) the alpha 4 and beta 2 subunits individually. The presence of the alpha 4 and beta 2 genes in the various baculovirus-infected Sf9 cells was confirmed following polymerase chain reaction (PCR) of the extracted viral DNAs, gel electrophoresis, and double strand sequencing. Autofluorography, following sodium dodecyl sulfate-polyacrylamide gel electrophoresis of infected cell lysates radiolabeled with 35S-methionine and immunoprecipated with mAb 270 (specific for the beta 2 subunit), revealed the presence of characteristic 52-kD bands in beta 2- and alpha 4 beta 2 recombinant viral-infected cells, but not in control cells or cells infected with wild-type virus or recombinant virus containing alpha 4 alone. The 52-kD protein, which is specific for mAb 270, is known to be the beta 2 subunit of neuronal nAChRs. Specific [3H]methylcarbamylcholine binding was observed in cells infected with both alpha 4 or beta 2 but not with the alpha 4 or beta 2 genes alone. Scatchard analysis revealed a Bmax = 5.50 pmol/mg and a Kd = 1 nM. The degree of [3H]methylcarbamylcholine binding/mg membrane protein was 180-fold greater than that found in rat brain. The study demonstrates that the major neuronal nAChR, which comprises alpha 4 and beta 2 subunits and is present in very low abundance in mammalian brain, can be prepared by a baculovirus expression system in sufficient quantities for chemical and crystallographic structural analysis.
Animals↗
Molecular and functional identification of m1 muscarinic acetylcholine receptors in rat ventricular myocytes.
The expression of muscarinic acetylcholine receptor (mAChR) subtypes in freshly isolated adult rat ventricular myocytes was investigated by reverse transcription of cellular mRNA followed by amplification of cDNA using the polymerase chain reaction (PCR). After reverse-transcriptase PCR, bands were obtained corresponding to the expected sizes for the m1 and m2 but not for the m3 to m5 mAChRs. The identity of the m1 and m2 bands was confirmed by single-cell PCR, restriction digest mapping, and Southern blot analysis. The presence of m1 and m2, but not m3, mAChR protein in these cells was shown by indirect immunofluorescence studies using subtype-specific antibodies. It was further investigated whether the identified m1 mAChR was responsible for the stimulatory effects on Ca2+ transients by high concentrations of carbachol ( > 10 mumol/L) known to occur in these cells. In pertussis toxin-treated ventricular myocytes electrically stimulated at 1 Hz, carbachol (300 mumol/L) increased the basal Ca2+ level from 96 +/- 7 to 118 +/- 8 nmol/L and the peak Ca2+ transient level from 519 +/- 32 to 640 +/- 36 nmol/L (mean +/- SEM P < .05 for both, n = 8). These effects of carbachol on Ca2+ transients were antagonized by 10 nmol/L pirenzepine, an m1 mAChR-selective antagonist. In contrast, the m2 mAChR-selective antagonist methoctramine (up to 100 nmol/L) did not inhibit the response. These results are the first to use single-cell PCR to probe cardiomyocyte-specific gene expression and indicate that m1 mAChRs are expressed on adult rat ventricular myocytes in addition to m2 mAChRs. The results further suggest that m1 mAChRs mediate the stimulatory responses on Ca2+ transients to high concentrations of cholinergic agonists seen in these cells.
Animals↗
3,3',4,4'-tetrachloroazobenzene absorption, disposition, and metabolism in male Fischer 344 rats.
3,3',4,4'-Tetrachloroazobenzene (TCAB) is a contaminant generated during the synthesis of 3,4-dichloroaniline and 3,4-dichloroaniline-derived pesticides. TCAB is isosteric to 2,3,7,8-tetrachlorodibenzo-p-dioxin and has been shown to bind to the Ah receptor. Following oral administration of [14C]TCAB (3.2 and 32 mg/kg), 39-45% of the dosed radioactivity was excreted into the urine and 53-56% was recovered in the feces within 48 hr. Less than 6% of the dosed radioactivity remained in the tissues examined at 96 hr. After intravenous administration (3.2 mg/kg), 33% of the dose was excreted in the bile during 6 hr. TCAB metabolites in urine were identified using LC/MS. The major metabolites were sulfate ester conjugates of hydroxylated mono- or dichloroaniline derivatives. Some of these metabolites were also acetylated. After intravenous administration, the disappearance of [14C]TCAB from blood was monitored, and the pharmacokinetic profile was consistent with a two-compartment model. Pharmacokinetic parameters reveal that the compound is readily cleared from the blood with a t1/2 of 4.0 hr, clearance of 12.3 ml/min.kg, and an apparent volume of distribution of 4.3 liters/kg. The absolute oral bioavailability was determined to be 30%. The extensive azo reduction of TCAB decreases its systemic absorption after oral administration and thereby limits the amount of parent compound available to interact with the Ah receptor and decreases the Ah receptor-mediated toxicity.
Administration, Oral↗
[Synthesis and activities of fragments of inhibin alpha subunit].
For further investigation of inhibin which is available only by laborious isolation, four fragments of inhibin alpha subunit (7-28, 37-65, 1-32, Y-1-32) were prepared by solid phase peptide synthesis using the Fmoc strategy. The effects of these fragments on progesterone production of rat corpus luteal (CL) cells in vitro were examined. The results indicate that these four fragments induced significant decrease of basic progesterone production.
Amino Acid Sequence↗