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Biomedical subjects

L Kang

Publications and source records attributed to L Kang.

At least 37 records · Page 2Linked to original sources

[Analysis of instability and deletion on chromosome 3p14 in Peutz-Jeghers syndrome].

OBJECTIVE: To determine the region where the tumor susceptible genes located, and to study chromosome instability, deletion and rearrangement of genes in Peutz-Jeghers Syndrome. METHODS: The method of induction was used to detect chromosome fragile sites of patients, Who were contrasted to familiar members and healthy individuals. DNA was extracted from leukocyte, polypus and tumor tissues, fresh or embedded in paraffin. The highly frequent abnormality of chromosome 3p14 region was studied with PCR, PCR-SSCP, microsatellite analysis, and DNA sequencing. RESULTS: The highest frequency of fragile site was in chromosome 3p14-3p24 region. The aberration frequency of D3S3340 is 21.4%, and that of D3S30 is 14.3%. Microsatellite analysis demonstrated LOH, of which the frequency is 42.9%; in addition, APC, k-ras mutation also occurred. The loss of guanylic acid was found in D3S1766. CONCLUSION: PJS has high chromosome instability, with deletion, mutation and arrangement of genes in 3p14 region, suggesting that cancer susceptible genes would be harbed in 3p14 cloning.

Adolescent↗

[The inhibitory effect of melatonin on morphine withdrawal syndromes and serum monoamines in morphine dependent mice].

OBJECTIVE: To observe the inhibitory effect of melatonin on morphine withdrawal syndromes and serum monoamines in morphine dependent mice. METHODS: A physical dependent model in mice was established by subcutaneous injection of morphine. The intensity of withdrawal syndromes was evaluated according to the jumping latency and jumping times. The concentration of serum monoamines was detected with HPLC-ECD. RESULTS: The physical withdrawal syndromes in morphine dependent mice were inhibited partly by four different doses (25, 50, 100, 200 mg/kg) of melatonin and showed a significant dose-dependent manner. The increased concentration of serum norepinephrine and dopamine in morphine-dependent mice could be reduced by large dose (100 mg/kg) of melatonin. CONCLUSION: The jumping withdrawal syndromes and serum monoamiues in morphine-dependent mice could be inhibited partly by melatonin.

Animals↗

Role of cardiac troponin I in the evaluation of myocardial injury.

Cardiac troponin I (cTnl) is highly specific for cardiac muscle. In this study, we compared the utility of CK and CK-MB index versus cTnl in the assessment of myocardial infarction in 155 patients being evaluated for myocardial damage. As a cardiac marker for MI, Troponin I seems to be superior to CK-MB. In the subset of patients with renal disease, cTnl has definite advantages over CK-MB. In addition, the use of cTnl has the potential to replace the measurement of lactate dehydrogenase isoenzymes.

Biomarkers↗

Vinculin is associated with the E-cadherin adhesion complex.

Cadherins mediate calcium-dependent cell-cell adhesion, and this activity is regulated by cytoplasmic interactions between cadherins, catenins, and the actin-based cytoskeleton. alpha-Catenin plays a critical role in the transmembrane anchorage of cadherins, and deletion of alpha-catenin has been shown to inactivate cadherin-mediated adhesion, resulting in a nonadhesive phenotype. Here we show that serum starvation increases E-cadherin expression and induces E-cadherin-dependent adhesion in the MDA-MB-468 breast cancer cell line. This adhesion occurred despite a lack of alpha-catenin expression, which was caused by mutations in the alpha-catenin gene. Coprecipitation analysis suggests that this adhesion may be mediated by cytoplasmic connections from cadherins to the cytoskeleton involving vinculin. A high level of vinculin associated with E-cadherin immunoprecipitates was observed in MDA-MB-468 cells. In contrast, vinculin was not detected in E-cadherin complexes in the A431 and MCF-7 epithelial carcinoma cell lines, which express alpha-catenin. However, in reciprocal immunoprecipitations using anti-vinculin antibodies, E-cadherin associated strongly with vinculin in MDA-MB-468 cells and, to a lesser extent, in A431 and MCF-7 cells. These results suggest that both alpha-catenin and vinculin may be present in the adhesion complex. To test the hypothesis that vinculin associates with E-cadherin complexes via beta-catenin, excess recombinant beta-catenin or alpha-catenin fusion protein was added to MDA-MB-468 cell lysates. Both specifically inhibited the coprecipitation of E-cadherin with vinculin, suggesting competition for the same binding site. These results suggest that vinculin plays a role in the establishment or regulation of the cadherin-based cell adhesion complex by direct interaction with beta-catenin.

Amino Acid Sequence↗

Fatty acid signals in Bacillus megaterium are attenuated by cytochrome P-450-mediated hydroxylation.

In previous publications [English, Hughes and Wolf (1994) J. Biol. Chem. 269, 26836-26841; English, Hughes and Wolf (1996) Biochem. J. 316, 279-283], we have demonstrated that peroxisome proliferators and non-steroidal anti-inflammatory drugs are inducers of the cytochrome P-450BM-3 gene in Bacillus megaterium ATCC14581. Their mechanism of action involves binding to and subsequent displacement of the transcriptional repressor, Bm3R1, from its operator site, which results in the activation of cytochrome P-450BM-3 gene transcription. We now present evidence that the branched-chain fatty acid, phytanic acid, is a potent inducer of cytochrome P-450BM-3. We have also observed that phytanic acid and peroxisome proliferators are inducers of Bm3R1 protein accumulation and associated DNA-binding activity. In contrast, several barbiturates, although capable of inducing cytochrome P-450BM-3 and Bm3R1 gene transcription, were unable to induce the Bm3R1 protein. We also demonstrate that cytochrome P-450BM-3 readily oxidizes phytanic acid, and provide evidence that, although the omega-1 hydroxy acid derivatives of phytanic acid can associate with Bm3R1, they do so with an affinity two orders of magnitude lower than the unmodified fatty acid. As a consequence, the ability of the hydroxylated product to induce cytochrome P-450BM-3 gene expression in vivo is markedly reduced. These data collectively suggest that metabolism of fatty acids by cytochrome P-450BM-3 leads to an attenuation of their ability to activate the transcription of the BM-3 operon. This work places the action of bacterial fatty acid hydroxylases in an autoregulatory loop where they may be responsible for the inactivation or clearance of the inducing fatty acid signal.

Bacillus megaterium↗

Mutation analysis of the putative tumor suppressor gene PTEN/MMAC1 in primary breast carcinomas.

A novel gene was identified recently at chromosome 10q23, named PTEN or MMAC1, and based on several criteria it was designated as a potential human tumor suppressor gene. Loss of heterozygosity affecting this region of 10q is observed in several cancer types, especially glioblastoma, and inactivating mutations of the PTEN/MMAC1 gene are found in some of these cancers as well as cell lines and xenografts. Breast cancer is among the tumor types in which mutations are documented, and germline mutations of the gene appear to be responsible for the rare autosomal dominant familial cancer syndrome known as Cowden disease, which includes breast cancer among its clinical features. To further determine the role that PTEN/MMAC1 mutations may play in breast tumorigenesis, the entire coding region was screened for mutations in 54 unselected primary breast cancers. Two mutations were identified, a somatic 2-bp deletion in an apparently sporadic breast cancer, and a germ-line 4-bp deletion in a breast cancer patient with a clinical history consistent with Cowden disease. These data indicate that somatic mutations of PTEN/ MMAC1 occur in only a small fraction of primary breast cancers and confirm the role of this gene in the etiology of Cowden disease. Evidence is also presented suggesting that numerous polymorphisms and missense variants exist in the PTEN/MMAC1 transcript.

Aged↗

Ribozyme-mediated high resistance against potato spindle tuber viroid in transgenic potatoes.

A hammerhead ribozyme [R(-)] targeting the minus strand RNA of potato spindle tuber viroid (PSTVd) and a mutated nonfunctional ribozyme [mR(-)] were designed, cloned, and transcribed. As predicted, both monomer and dimer transcripts of the active R(-) ribozyme gene could cleave the PSTVd minus strand dimer RNA into three fragments of 77, 338, and 359 bases in vitro at 25 and 50 degrees C. The tandem dimer genes of R(-) and mR(-) were subcloned separately into the plant expression vector pROK2. Transgenic potato plants (cultivar Desirée) were generated by Agrobacterium tumefaciens-mediated transformation. Twenty-three of 34 independent transgenic plant lines expressing the active ribozyme R(-) resulted in having high levels of resistance to PSTVd, being free of PSTVd accumulation after challenge inoculation with PSTVd, but the remaining lines showed weaker levels of resistance to PSTVd with low levels of PSTVd accumulation. In contrast, 59 of 60 independent transgenic lines expressing the mutated ribozyme mR(-) were susceptible to PSTVd inoculation and had levels of PSTVd accumulation similar to that of the control plants transformed with the empty vector. The resistance against PSTVd replication was stably inherited to the vegetative progenies.

Base Composition↗

Studies on the mechanism underlying the antifibrillatory effect of the A1-adenosine agonist, R-PIA, in rat isolated hearts.

Previous studies in pigs have shown that the A1-adenosine receptor agonist, R-PIA, is a potent antifibrillatory agent during myocardial ischaemia and that this effect can be overriden by atrial pacing. However, reports of A1-adenosine receptor down-regulation following chronic exposure to A1-receptor agonists suggest that this may limit their use as potential antiarrhythmic therapy. The acute and chronic effects of R-PIA were examined on ventricular arrhythmias and hemodynamics in Langendorff-perfused rat isolated hearts subjected to acute regional myocardial ischemia in an attempt to confirm the heart rate dependency of the antifibrillatory mechanism of R-PIA and to assess the effects of chronic treatment on this protection. Acute challenge with R-PIA (10(-10) to 5 x 10(-8) M; n = 10 for all groups) produced a concentration-dependent bradycardia prior to coronary occlusion. Coronary artery occlusion in control hearts (n = 20) resulted in an immediate increase in perfusion pressure, from 61 +/- 6 to 87 +/- 7 mmHg within 5 minutes, followed by a gradual continued rise reaching a maximum of 123 +/- 9 mmHg by the end of the 30-minute experimental period. R-PIA significantly attenuated the sustained increase in perfusion pressure in a non-concentration-dependent manner. A concentration of 10(-10) M R-PIA had no effect on the incidence of ventricular fibrillation (VF), while all higher concentrations reduced the incidence of VF to a similar degree (from 60% in controls to 10%, 20%, 10%, and 0% with 10(-9), 5 x 10(-9), 10(-8), and 5 x 10(-8) M R-PIA, respectively). R-PIA also reduced the total ventricular premature beat (VPB) count, but in a concentration-dependent manner. Chronic treatment of the rats with R-PIA (50 microg/kg i.p., bd; n = 10) for 7 days caused a significant attenuation of the bradycardic response to acute perfusion in vitro with R-PIA (10(-8) M) and abolished the attenuation of the sustained rise in perfusion pressure during myocardial ischemia. The antifibrillatory effect of R-PIA, however, was unaffected by chronic pretreatment (VF incidence 0% vs. 70% in control hearts from rats that had been given chronic saline ip injections n = 10; P < 0.01). These results suggest that the bradycardia induced by acute R-PIA may not be the mechanism underlying the antifibrillatory effect of R-PIA, while the reduction in the less severe arrhythmias is heart rate dependent. Furthermore, while chronic treatment with R-PIA significantly attenuates the heart rate response to acute R-PIA challenge, the antifibrillatory properties remain intact.

Adenosine↗

N-cadherin promotes adhesion between invasive breast cancer cells and the stroma.

Calcium-dependent cell adhesion molecules (cadherins) are involved in maintaining the epithelial structure of a number of tissues including the mammary gland. In breast and other tumor types, loss of E-cadherin expression has been seen in high grade tumors and correlates with increased invasiveness. Here we show high levels of expression of N-cadherin in the most invasive breast cancer cell lines which was inversely correlated with their expression of E-cadherin. A stromal cell line also expressed N-cadherin in accordance with its fibroblastic morphology. N-cadherin localized to areas of cell-cell contact in all cells that expressed it. Calcium-dependent intercellular adhesion of N-cadherin-expressing breast cancer and stromal cells was specifically inhibited by an anti N-cadherin monoclonal antibody. In addition, N-cadherin promoted the interaction of invasive breast cancer cells with mammary stromal cells; in contrast, E-cadherin expressing cell lines did not co-aggregate with stromal cells. The combined results suggest a functional role for N-cadherin in cohesion of breast tumor cells which, in addition promotes their interaction with the surrounding stromal cells, thereby facilitating invasion and metastasis.

Animals↗

Morphine activates opioid receptors without causing their rapid internalization.

We have examined the endocytic trafficking of epitope-tagged delta and mu opioid receptors expressed in human embryonic kidney (HEK) 293 cells. These receptors are activated by peptide agonists (enkephalins) as well as by the alkaloid agonist drugs etorphine and morphine. Enkephalins and etorphine cause opioid receptors to internalize rapidly (t1/2 approximately 6 min) by a mechanism similar to that utilized by a number of other classes of receptor, as indicated by localization of internalized opioid receptors in transferrin-containing endosomes and inhibition of opioid receptor internalization by hypertonic media. Remarkably, morphine does not stimulate the rapid internalization of either delta or mu opioid receptors, even at high concentrations that strongly inhibit adenylyl cyclase. These data indicate that agonist ligands, which have similar effects on receptor-mediated signaling, can have dramatically different effects on the intracellular trafficking of a G protein-coupled receptor.

Cell Line↗

156 cases of Gilles de la Tourette's syndrome treated by acupuncture.

The principle of clearing Yangming and nourishing the kidney and heart was adopted in the treatment of 156 cases of Gilles de la Tourette's Syndrome with acupuncture. The total effective rate was 92.3%, and the cure rate in children aged 11-15 years was markedly higher than that in children 6-10 years of age. Among 84 cases with abnormal EEG, the pathological waves in 54 disappeared or ameliorated after the treatment.

Acupuncture Therapy↗

Androgen biosynthesis and secretion in developing Xenopus laevis.

In the frog, Xenopus laevis, castration or anti-androgen treatment during late tadpole or early juvenile stages blocks masculinization of vocal neuroeffectors in males while exogenous androgen or a testicular transplant masculinizes vocal neuroeffectors in females. To elucidate the relation between androgen secretion and the process of masculinization, we measured androgens by radioimmunoassay in adults, juveniles, and tadpoles. In adult females, values for both testosterone (T) and dihydrotestosterone (DHT) were essentially identical during the winter and the summer while in males, summer values for both androgens were 7 to 8x winter values. For DHT, circulating values in males significantly exceeded those in females in both winter and summer while for T, values differed significantly only in summer. Sex differences in circulating androgen arise at late juvenile stages (PM4 and PM5); T values are higher in males than in females and higher, for both sexes, in winter than in summer. During early juvenile stages (PM1-PM3), there were no seasonal or sex differences in circulating androgens. While androgens were detected in liver during late tadpole stages (stage 56-66), no sex differences were apparent. Androgens can be detected in tadpole tissues, as early as tadpole stage 47. delta 5-3 beta-Hydroxysteroid dehydrogenase (HSD) histochemistry indicates that tadpole gonads and interrenals (and, to a much lesser extent, kidneys) are the only tissues capable of steroid biosynthesis. Interrenal HSD activity was present throughout tadpole development; in gonads, activity began before sexual differentiation. Thin-layer chromatographic separation of steroid metabolites following in vitro incubation with radioactive precursors indicated that the interrenals were more active than the gonads in tadpoles but not in juveniles. Thus, interrenals and gonads of developing X. laevis can synthesize delta 4 steroids (which include androgens) and, from early tadpole stages, are capable of metabolizing pregnenolone and testosterone, early and late compounds in the steroid synthetic pathway, respectively. The absence of dramatic sex differences in T or DHT levels during late tadpole and early juvenile stages suggests that while necessary, androgen secretion is not sufficient for masculinization of vocal neuroeffectors.

Aging↗

Structure and function of ASP, the human homolog of the mouse agouti gene.

The mouse agouti coat color gene encodes a novel paracrine signaling molecule whose pulsatile expression produces a characteristic pattern of banded pigment in individual hairs. Several spontaneous agouti alleles produce adult-onset obesity and diabetes, and have provided important single-gene animal models for alterations in energy metabolism. Utilizing linkage groups conserved between mice and humans, we have cloned the human homolog of the mouse agouti gene from a human chromosome 20 yeast artificial chromosome known to contain S-adenosyl homocysteine hydrolase (AHCY). The human agouti gene, named Agouti Signaling Protein (ASP), encodes a 132 amino acid protein, the mRNA for which is expressed in testis, ovary, and heart, and at lower levels in liver, kidney, and foreskin. As predicted by the interactions of mouse agouti with the extension gene (which encodes the melanocyte receptor for alpha-melanocyte stimulating hormone [alpha-MSH]), expression of ASP in transgenic mice produces a yellow coat, and expression of ASP in cell culture blocks the alpha-MSH-stimulated accumulation of cAMP in mouse melanoma cells. The localization of ASP relative to other loci on chromosome 20 excludes it as a candidate for the MODY1 locus, a gene responsible for one form of early-onset non-insulin-dependent diabetes mellitus or maturity-onset diabetes of the young. The expression of ASP in human tissues suggests a function for agouti homologs in species that do not exhibit the characteristic phenotype of banded hairs.

Agouti Signaling Protein↗

Nucleotide sequence of maize dwarf mosaic virus capsid protein gene and its expression in Escherichia coli.

The 3'-terminal 1,279 nucleotide sequence of maize dwarf mosaic virus (MDMV) genome has been determined. This sequence contains an open reading frame of 1,023 nucleotides and a 3'-non-coding region of 256 nucleotides. The open reading frame includes all of the coding regions for the viral capsid protein (CP) and part of the viral nuclear inclusion protein (NIb). The predicted viral CP consists of 313 amino acid residues with a calculated molecular weight of 35,400. The amino acid sequence of the viral CP derived from MDMV cDNA shows about 47%-54% homology to that of 4 other potyviruses. The viral CP gene was constructed in frame with the lacZ gene in pUC19 plasmid and expressed in E. coli cells. The fusion polypeptide positively reacted in Western blot with an antiserum prepared against the native viral CP.

Amino Acid Sequence↗

The effect of single bolus dose of esmolol for controlling the tachycardia and hypertension during laryngoscopy and tracheal intubation.

Tachycardia and hypertension usually accompany laryngoscopy and tracheal intubation. This response is undesirable, especially in patients with cardiovascular or intracranial diseases. Esmolol is a cardioselective, ultrashort-acting beta adrenergic blocking agent with a very short half-life. The efficacy of bolus dose of esmolol in blunting hemodynamic responses during laryngoscopy and tracheal intubation was evaluated. 45 patients (15 in each group) of ASA physical status I and II scheduled for elective non-cardiac surgery were included in this randomized, placebo-controlled study. At time zero, the study preparation (placebo, 100 or 200 mg of esmolol) was administered intravenously, followed by thiopentone 5 mg/kg and succinylcholine 1.5 mg/kg for induction. Tracheal intubation was performed 2 minutes after time zero. Anesthesia was maintained with 50% nitrous oxide and 1.0 MAC halothane in oxygen, and vecuronium 0.08 mg/kg. Heart rate (HR) and systolic blood pressure (SBP) were recorded every minute for 10 minutes. To compare with the placebo group, there was a significant decrease in either HR or SBP in 200 mg group in the 8 minutes course after intubation (p < 0.05). There was a significant decrease in HR in the 100 mg group at the 3rd, 4th, and 5th minutes when compared with the placebo group (p < 0.05). The differences in SBP between the 100 mg group and placebo group were significant at the 3rd and 4th minutes (p < 0.05). Both bolus dosages of esmolol could effectively attenuate the tachycardia and hypertension produced by laryngoscopy and tracheal intubation. Furthermore, esmolol 200 mg presented a better hemodynamic stability than esmolol 100 mg during induction of anesthesia.

Adrenergic beta-Antagonists↗

[Serology in patients with scleroderma].

In order to evaluate the practical clinical value of centromere, Scl-70, and nucleolar antibodies as demonstrated routinely by the Autoimmune Department of the Serum Institute of Copenhagen, 1293 sera from 497 patients with scleroderma (SSc) and other connective tissue diseases were tested for the three antibodies and for other nuclear antibodies. The three antibodies were found in 32, 15 and 15%, respectively, of sera from patients with SSc. Since more than one of the three antibodies was rarely demonstrated in any one serum, one of them was found in two thirds of sera from patients with SSc. The specificity of the three antibodies for SSc was 95% or more. Centromere antibody was found most frequently in patients with limited SSc. Scl-70 antibody was found almost exclusively in sera from patients with extensive SSc (involving the skin of the trunk). In such sera, centromere antibody was found in only 21%. Scl-70 antibody was overrepresented and centromere antibody was underrepresented in sera from patients with pulmonary involvement, the converse being true for sera from patients with calcinosis, esophageal involvement and telangiectasia.

Antibody Specificity↗

The use of transesophageal echocardiography to evaluate the effectiveness of patent ductus arteriosus ligation.

The ligation of patent ductus arteriosus (PDA) is a comparatively easy operation, but some complications are possible. The most common complication is incomplete ligation of the PDA; others include inadvertent ligation of the descending aorta or left pulmonary artery, transient rise in systemic blood pressure and increased left ventricular afterload, and acute right heart failure due to pulmonary hypertension. The completeness of the PDA ligation is usually determined only by the operating physician's experience, including the use of an esophageal stethoscope or a finger on the lesion to feel for vibration. These methods sometimes fail to detect an incomplete ligation. With transesophageal echocardiography (TEE), we have monitored the entire course of the PDA ligation directly without interrupting the surgical procedure, and precisely determined the completeness of the ligation. We also expect that TEE will enable us to avoid other complications as well.

Adolescent↗

[The effects of intravenous and intraperitoneal dexamethasone treatment on vecuronium in the anesthetized rat].

It is fairly widespread clinical practice to administer large doses of corticosteroids to patients in cases of shock; doses of hydrocortisone as high as 50 mg/kg given intravenously have been proposed and used; especially in cases of Myasthenia Gravis. Its effects on neuromuscular transmission is not yet fully understood. In order to determine the mechanism of this interaction, we undertook this investigation. The neuromuscular effects of Dexamethasone, at single dose of 100 micrograms/kg intravenously or intraperitoneally, were studied by electromyographical quantification of the tibialis-anterior muscle evoked by sciatic nerve stimulation in 21 rats anesthetized with Urothane 1.25 g/kg given intraperitoneally. Intravenous Dexamethasone (n = 7) had no obvious effect on the blockade of tibialis-anterior muscle produced by the cumulative doses of Vecuronium (ED50 = 180 +/- 24 micrograms/kg), as compared with the control group (n = 7) (ED50 = 201 +/- 18 micrograms/kg). In contrast, intraperitoneal Dexamethasone (n = 7) produced a significant (P < 0.05) change (ED50 = 240 +/- 28 micrograms/kg), when compared with control group. The results showed that in tibialis-anterior muscle sciatic nerve preparation of rats, intraperitoneal Dexamethasone had antagonism effect on Vecuronium. However, there was no obvious influence on Vecuronium in the intravenous Dexamethasone group.

Anesthesia↗