Biomedical subjects

E C Chan

Publications and source records attributed to E C Chan.

At least 37 records · Page 2Linked to original sources

PubMed2000

Detection of mRNA of sodium iodide symporter in benign and malignant human thyroid tissues.

Human sodium iodide symporter (hNIS) is an intrinsic membrane protein with 12 transmembrane regions, which shows homology to other sodium-dependent transporters. There is controversy as to the amount of hNIS expression in different kinds of human thyroid cancer tissues and cell lines. In this study, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect mRNA of hNIS in various fresh normal, benign tissues and malignant human thyroid tissues. The forward primer was nested hNIS-5' primer containing the sequences: ACCTGGAAATGCGCTTCAGC. The reverse primer was nested hNIS-3' primer containing the sequences: AAGCATGACACCGCGTGCCA. The results revealed three of three normal tissues, six of eight nodular hyperplasia, two of two hyperthyroidism, one of three follicular adenomas, five of ten papillary thyroid carcinomas, one of one follicular carcinoma and zero of one metastatic follicular tissues demonstrated positive results for hNIS in thyroid epithelial cells. A higher percentage of positive results of the symporter mRNA were found in normal benign thyroid tissues and the thyroid tissues of hyperthyroidism, and nodular hyperplasia (84.6%); however follicular adenoma, papillary and follicular thyroid carcinomas demonstrated a lower percentage of expression in the RT-PCR studies (46.7%). Serum thyrotropin levels and the degree of differentiated components presented in cancer tissues have been mentioned as important factors for hNIS expression in the cancer tissues. The discrepancies of the expression of hNIS in in vivo and in vitro studies need further investigation. In conclusion, hNIS was found in higher ratios in normal and benign thyroid tissues than in the malignant tissues. In addition, the RT-PCR technique hNIS did not detect the transporter in most papillary thyroid cancer tissues.

Adenocarcinoma, Follicular↗

PubMed2000

Purification and characterization of neutral sphingomyelinase from Helicobacter pylori.

Phospholipase activities of human gastric bacterium, Helicobacter pylori, are regarded as the pathogenic factors owing to their actions on epithelial cell membranes. In this study, we purified and characterized neutral sphingomyelinase (N-SMase) from the superficial components of H. pylori strains for the first time. N-SMase was purified 2083-fold with an overall recovery of 37%. The purification steps included acid glycine extraction, ammonium sulfate precipitation, CM-Sepharose, Mono-Q, and Sephadex G-75 column chromatography. Approximate molecular mass for the native N-SMase was around 32 kDa. When N-omega-trinitrophenylaminolauryl sphingomyelin (TNPAL-SM) was used as a substrate, the purified enzyme exhibited a K(m) of 6.7 microM and a V(max) of 15.6 nmol of TNPAL-sphingosine/h/mg of protein at 37 degrees C in 50 mM phosphate-buffered saline, pH 7.4. N-SMase reaches optimal activity at pH 7.4 and has a pI of 7.15. The enzyme activity is magnesium dependent and specifically hydrolyzed sphingomyelin and phosphatidylethanolamine. The enzyme also exhibits hemolytic activity on human erythrocytes. According to Western blot analysis, a rabbit antiserum against purified N-SMase from H. pylori cross-reacted with SMase from Bacillus cereus. Sera from individuals with H. pylori infection but not uninfected ones recognizing the purified N-SMase indicated that it was produced in vivo. In enzyme-linked immunosorbent assays, the purified N-SMase used as an antigen was as effective as crude protein antigens in detecting human antibodies to H. pylori.

Antibodies, Bacterial↗

PubMed2000

Expression of capsid [correction of caspid] protein VP1 for use as antigen for the diagnosis of enterovirus 71 infection.

To produce enterovirus 71 antigen for diagnostic purposes, the gene encoding the entire capsid protein VP1 was amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned and expressed in Escherichia coli as a poly-histidine fusion protein. Western blotting experiments with sera from patients with enterovirus 71 infection indicated that immunoglobulin G (IgG) and IgM antibodies bound to a single polypeptide VP1. According to these results, IgM anti-VP1 appeared in sera of patients with a symptomatic enterovirus 71 acute infection, whereas IgG anti-VP1 was present in sera of past infection. This finding suggests that detecting IgG and IgM immune responses against linear epitopes of recombinant VP1 is an effective means of determining the different phases of enterovirus 71 infection. In addition, sera containing coxsackie virus 16 (CA16) antibodies did not cross-react with the recombinant VP1 of enterovirus 71, despite the homology between VP1 proteins of both viruses. Comparison with reference PCR and neutralization assays showed these antibody tests to be appropriate for the serodiagnosis of enterovirus 71 infection.

Antibodies, Viral↗

PubMed2000

High-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometric method for the analysis of catecholamines and metanephrines in human urine.

An assay of norepinephrine (NE), epinephrine (E), dopamine (DA), normetanephrine (NE) and metanephrine (MN) based on high-performance liquid chromatography (HPLC) in combination with atmospheric pressure chemical ionization mass spectrometry (APcI-MS) is described. The catecholamines and metanephrines were extracted from urine and aqueous samples using Bio-Rex 70 cation-exchange resin and subjected to analysis by HPLC/APcI-MS. The separation was performed on a C18 column in 50 mM ammonium formate buffer, pH 3.0, using a flow rate of 0.8 mL/min. Acetonitrile was added post-column at a flow rate of 0.2 mL/min via a post-column addition tee. The total analysis time was 6.5 min. The quantitative analysis was performed using 3,4-dihydroxybenzylamine (DHBA) as the internal standard (I.S.). Selected ion monitoring detection was applied: m/z 170 (for NE), 184 (for E and NM), 154 (for DA), 198 (for MN) and 140 (for DHBA, I.S.). The limits of quantitation were 5 ng/mL for NE, E and DA and 2.5 ng/mL for NM and MN. The recovery ranged from 75 to 83% for each analyte. The method was found to be simple and highly selective for the determination of catecholamines and metanephrines in the urine of patients suspected of pheochromocytoma.

Atmospheric Pressure↗

PubMed2000

Clinical outcomes of post-operative locoregional radiotherapy in pre-menopausal and post-menopausal Chinese women with breast cancer.

BACKGROUND AND PURPOSES: Recent randomized studies from the West show that post-operative locoregional radiotherapy improves survival in lymph node (LN) positive pre-menopausal women with breast cancer but this benefit has not been established in the Chinese population. There is no published study on clinical outcomes (locoregional recurrence, survival and toxicities) of post-operative locoregional radiotherapy in Chinese women with breast cancer. MATERIALS AND METHODS: We conducted a retrospective study of 399 female Chinese patients with breast cancer who had received post-mastectomy locoregional radiotherapy in our center between 1984 and 1990. The patients were stratified according to tumor size, menopausal and LN status. We analyzed the incidence and pattern of locoregional recurrence, distant recurrence, survival rates and toxicity related to radiotherapy. RESULTS: Of the 399 patients 216 were pre-menopausal and 183 were post-menopausal. The mean age was 49.3 years (23-83). Distribution of tumor size and LN status of the two groups was similar. Median follow-up was 71.9 months. Locoregional recurrence occurred in 57 (14.3%) patients (pre-menopausal 24 (11.1%); post-menopausal 33 (18.3%) P=0.489). Recurrence was more common in LN positive patients (18.2%) than LN negative patients (9.2%). The pattern of locoregional recurrence was: chest wall 24, axilla LN 12, supraclavicular LN 10, chest plus LN 11. The distant recurrence rate was 39.6% for all patients and 75.4% for patients with locoregional recurrences. The overall 10-year survival rate for all patients was 54%. For LN positive patients the 10-year survival rate of pre-menopausal and post-menopausal women was 38 and 51%, respectively (P=0.207), and for LN negative patients the rate was 73 and 70%, respectively (P=0.603). Acute skin toxicity included redness (30.8%), dry desquamation (12.8%), and wet desquamation (6. 8%). Long-term toxicities included skin atrophy (0.3%), telangectasia (3.3%), pneumonitis (2.8%) and brachial plexus palsy (1.3%). CONCLUSIONS: In our series Chinese patients with node-positive breast cancer have a relatively high locoregional recurrence rate in spite of mastectomy and post-operative radiotherapy. Limited use of adjuvant system chemotherapy may account, at least in part, for this finding. Clinical outcomes of post-operative radiotherapy in pre-menopausal and post-menopausal breast cancer patients are similar.

Adult↗

PubMed2000

Evaluation of degradation of urinary catecholamines and metanephrines and deconjugation of their sulfoconjugates using stability-indicating reversed-phase ion-pair HPLC with electrochemical detection.

The possibility of the deconjugation of urinary catecholamine sulfates (CA-S) and metanephrine sulfates (MN-S) during storage was studied using a reversed-phase ion-pair HPLC method with electrochemical detection. Stability profiles of catecholamines (norepinephrine (NE), epinephrine (E) and dopamine (DA)) and metanephrines (normetanephrine (NM) and metanephrine (MN)) in preserved and unpreserved pooled urine and aqueous samples stored at 10 and 30 degrees C over 8 days were compared. Results showed that these compounds exhibited stable or declining profiles in aqueous samples but fluctuating profiles in pooled urine samples. It was concluded that deconjugation of CA-S and MN-S occurred in both preserved and unpreserved urine samples. Therefore, at 10 and 30 degrees C, levels of detected urinary catecholamines and metanephrines could be affected by both the degradation of the free amines and deconjugation of the CA-S and MN-S. Based on these findings, another batch of unpreserved urine and aqueous samples were prepared and stored at -80 degrees C. Under this condition, all compounds were found to be stable with less than 10% variations in both unpreserved urine and aqueous samples for at least 22 days. These indicated that urine samples could be stored unpreserved at -80 degrees C for at least 3 weeks without significant degradation of the free amines or deconjugation of their sulfoconjugates.

Adult↗

PubMed2000

Expression of sodium iodide symporter in metastatic and follicular human thyroid tissues.

Active iodide uptake across the basal membrane mediated by human sodium iodide symporter (hNIS) has been shown to be a process coupled with the flow of sodium. There is still controversy as to the amount of hNIS expression present in different kinds of human thyroid cancer tissues. In this study, we present a 58-year-old women with follicular thyroid carcinoma with vertebra and skull metastases. 201Tl and 5 mCi 131I scans clearly demonstrated the metastatic lesions in the brain of this patient. Thyroid and metastatic tissues were then obtained for this study, which is aimed at comparing the iodide trapping ability in vivo and in vitro of hNIS, and then comparing their expression in both thyroid tissue and metastatic tissues. Polyclonal antibodies to hNIS and competitive RT PCR were used to analyze the symporter protein and mRNA expressed in follicular human thyroid and metastatic tissues. Positive staining of the symporter protein was performed in the follicular thyroid carcinomas, otherwise, the metastatic tissues could not have demonstrated the protein in the staining. Follicular thyroid carcinoma tissues from thyroid were revealed around 5 pg hNIS expressed in follicular thyroid carcinoma tissues from the thyroid. Otherwise, there was almost an absence of hNIS expression in the metastatic tissue. These discrepancies of the expression in hNIS in vivo and in vitro studies need further investigation.

Adenocarcinoma, Follicular↗

PubMed2000

Taxonomy and virulence of oral spirochetes.

All oral spirochetes are classified in the genus Treponema. This genus is in the family Spirochaetaceae as in Bergey's manual of systematic bacteriology. Other generic members of the family include Spirochaeta, Cristispira and Borrelia. This conventional classification is in accord with phylogenetic analysis of the spirochetes based on 16S rRNA cataloguing. The oral spirochetes fall naturally within the grouping of Treponema. Only four species of Treponema have been cultivated and maintained reliably: Treponema denticola, Treponema pectinovorum, Treponema socranskii and Treponema vincentii. These species have valid names according to the rules of nomenclature except for Treponema vincentii, which only has had effective publication. The virulence factors of the oral spirochetes updated in this mini-review have been discussed within the following broad confines: adherence, cytotoxic effects, iron sequestration and locomotion. T. denticola has been shown to attach to human gingival fibroblasts, basement membrane proteins, as well as other substrates by specific attachment mechanisms. The binding of the spirochete to human gingival fibroblasts resulted in cytotoxicity and cell death due to enzymes and other proteins. Binding of the spirochete to erythrocytes was accompanied by agglutination and lysis. Hemolysis releases hemin, which is sequestered by an outer membrane sheath receptor protein of the spirochete. The ability to locomote through viscous environments enables spirochetes to migrate within gingival crevicular fluid and to penetrate sulcular epithelial linings and gingival connective tissue. The virulence factors of the oral spirochetes proven in vitro underscore the important role they play in the periodontal disease process. This role has been evaluated in vivo by use of a murine model.

Animals↗

PubMed2000

Two novel factor VII gene mutations in a Chinese family with factor VII deficiency.

We report two novel factor VII (FVII) gene mutations in a Chinese family with FVII deficiency. The proband, a 55-year-old woman. was incidentally found to have right shoulder arthritis consistent with chronic haemophilic arthropathy. FVII studies showed a FVII activity of 0.02 iu/ml and a FVII antigen of 49%. Molecular analysis showed a double heterozygous state, with an exon 4 nonsense mutation (C6003-->A; Cys61-->Term) and an exon 8 missense mutation (T10902-->G; Cys329-->Gly) that disrupted a Cys310/Cys329 disulphide bond. The genotypes and phenotypes were correlated in the patient's daughters. Two daughters were heterozygous for the Cys61-->Term mutation and showed a type 1 FVII gene mutation phenotype consistent with a nonsense mutation. One daughter was heterozygous for the Csy329-->Gly mutation and showed a type 2 mutation phenotype consistent with a missense mutation. These are the first reported FVII gene mutations in the Chinese people.

China↗

PubMed2000

Biodegradable alginate antibiotic beads.

The authors investigated the poly-L-lysine-coated alginate beads as an antibiotic delivery system for the treatment of various surgical infections. The sodium alginate was mixed with vancomycin, coated with poly-L-lysine, and lyophilized to form five types of the biodegradable antibiotic beads. Type I, 2.5% alginate, nonpoly-L-lysine coated and nonlyophilized; Type II, 2.5% alginate, poly-L-lysine coated but nonlyophilized; Type III, 2.5% alginate, poly-L-lysine coated and lyophilized; Type IV, 5% alginate, poly-L-lysine coated and lyophilized; and Type V, 7.5% alginate, poly-L-lysine coated and lyophilized. Cytotoxicity of the alginate beads to fibroblasts and HeLa cells was evaluated by the MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide] colorimetric assay. A study of in vitro elution of vancomycin of the alginate antibiotic beads was performed. The results suggested that the alginate antibiotic beads present no obvious toxic risk to their use as a drug delivery system. The concentration of vancomycin in these five types of beads was well above the breakpoint sensitivity concentration (the antibiotic concentration at the transition point between bacterial killing and resistance to the antibiotic) for 9,11,12, 14, and 17 days respectively. The release was most marked during the first 3 days. The duration of antibiotic release was prolonged by using techniques of poly-L-lysine coating, lyophilization, and by increasing the content of alginate. This study offers a biodegradable delivery system of antibiotics to treat various surgical infections.

Alginates↗

PubMed2000

Relaxation to flavones and flavonols in rat isolated thoracic aorta: mechanism of action and structure-activity relationships.

The mechanism of the relaxant action and the structure-activity relation of flavonols (fisetin, quercetin, and 3,3',4'-trihydroxyflavone) and flavones (apigenin, chrysin, and luteolin) were examined in rat isolated thoracic aorta. The control responses to flavonols and flavones were compared with responses observed after the removal of the endothelium or in the presence of the L-type Ca2+ channel blocker, nifedipine (10(-7) M). The effects of flavonoids on contraction caused by the influx of extracellular Ca2+ and agonist-induced release of intracellular Ca2+ also were investigated. The flavones exhibited endothelium-independent vasorelaxation, whereas the removal of the endothelium significantly decreased the sensitivity of the relaxant responses to the flavonols without affecting the maximal relaxation. In the presence of nifedipine, the responses to apigenin, luteolin, and quercetin were significantly inhibited, but relaxation to chrysin, fisetin, and 3,3',4'-trihydroxyflavone was unaffected. All flavonols and flavones caused concentration-dependent inhibition of the contractile responses to exogenous application of Ca2+ and the release of intracellular Ca2+ stimulated by phenylephrine. Of the six flavonoids examined, 3,3',4'-trihydroxyflavone was the most potent when causing vasorelaxation or inhibition of contraction caused by the influx or release of Ca2+. In conclusion, these studies provide evidence that the hydroxyl substitution in the carbon 3 position that characterizes the flavonols is important in stimulating endothelium-dependent vasorelaxation, and the absence of hydroxyl substitution on the A phenolic ring enhances the relaxant action.

Animals↗

PubMed2000

Lipophilic antioxidants and iron status in ESRD patients on hemodialysis.

Cardiovascular disease remains the major cause of mortality in hemodialysis patients. Abnormal oxidative stress and impaired antioxidant defense may contribute to accelerated atherogenesis associated with uremia. As oxidative modification of lipids appears to be a prerequisite for the development of atherosclerotic lesions, lipophilic antioxidants may be protective. The aim of this study was to determine the plasma levels of lipophilic antioxidants in 82 hemodialysis patients and 30 controls and to investigate the influence of body iron status on the levels of lipophilic antioxidants. The patients were categorized into 3 groups according to their serum ferritin levels. We found that the plasma levels of lycopene, delta-tocopherol, gamma-tocopherol and retinol of hemodialysis patients were lower than those of controls. On the other hand, both absolute and lipid-normalized plasma lycopene levels were significantly reduced in those patients in the groups with higher ferritin levels as compared to those with lower ferritin levels. In addition, our study showed that the lipid-normalized plasma levels of beta-carotene and alpha-carotene of hemodialysis patients with higher ferritin levels were lower than those of the patients with lower levels. These data suggest that the plasma levels of lipophilic antioxidants are altered in end-stage renal disease on hemodialysis and may be considered as markers of oxidative stress in these patients. Most importantly, elevated serum ferritin levels may affect the levels of these lipophilic antioxidants.

Adult↗

PubMed2000

Corticotropin-releasing hormone gene expression in primary placental cells is modulated by cyclic adenosine 3',5'-monophosphate.

CRH, the principal neuropeptide regulator of pituitary ACTH secretion, is also expressed in placenta. Placental CRH has been linked to the process of human parturition. However, the mechanisms regulating transcription of the CRH gene in placenta remain unclear. cAMP signaling pathways play important roles in regulating the expression of a diverse range of endocrine genes in the placenta. Therefore, we have explored the effect of cAMP on CRH promoter activity in primary cultures of human placental cells. Both forskolin and 8-bromo-cAMP, activators of protein kinase A, can increase CRH promoter activity 5-fold in transiently transfected human primary placental cells, in a manner that parallels the increase in endogenous CRH peptide. Maximal stimulation of CRH promoter activity occurs at 500 micromol/L 8-bromo-cAMP and 10 micromol/L forskolin. Electrophoretic mobility shift assay and mutation analysis combined with transient transfection demonstrate that in placental cells cAMP stimulates CRH gene expression through a cAMP regulatory element in the proximal CRH promoter region and involves a placental nuclear protein interacting specifically with the cAMP regulatory element.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed2000

Glucocorticoid stimulation of corticotropin-releasing hormone gene expression requires a cyclic adenosine 3',5'-monophosphate regulatory element in human primary placental cytotrophoblast cells.

Production of placental CRH, which is identical to the peptide synthesized and secreted in the hypothalamus, has been linked to human parturition. Glucocorticoids stimulate placental CRH secretion and messenger ribonucleic acid expression, in contrast to their inhibition of CRH synthesis in the hypothalamus. A positive feedforward loop involving glucocorticoid-CRH-ACTH-glucocorticoid is thought to drive the exponential increase in placental CRH leading to delivery. Tissue-specific effects of glucocorticoids on CRH expression are therefore of interest. Using human primary placental cells, we investigated the mechanism by which glucocorticoids stimulate placental CRH gene expression. Nuclear run-on transcription shows that in human placental cells glucocorticoids up-regulate transcription of human CRH (hCRH). Using transient transfection assays we demonstrate that dexamethasone up-regulates both basal and cAMP-stimulated hCRH promoter activity, correlating well with the increase in endogenous CRH peptide levels. Through mutagenesis and deletion analyses we show that dexamethasone stimulation of hCRH gene transcription requires a functional cAMP regulatory element (CRE); this CRE is adequate to confer dexamethasone stimulation upon a heterologous promoter, and electrophoretic mobility shift assay studies show that a placental nuclear protein specifically binds to the hCRH CRE.

Cell Nucleus↗

PubMed1999

Alzheimer's disease may not be a spirochetosis.

It has been reported previously that spirochetes could be one of the causes of Alzheimer's disease (AD). In this study, we have attempted to reproduce these findings by examining fresh blood samples from 22 patients diagnosed with early stage (n = 16) and late stage (n = 6) AD. The patients were participants in a clinical drug trial. Fresh necropsy brain cortical specimens from AD patients (n = 7) were also examined. Spirochetes were observed microscopically in the blood of only one late-stage AD patient. None of the brain tissues showed the presence of spirochetes. Our results suggest that spirochetes are probably not associated with AD.

Aged↗

PubMed1999

In vitro elution of vancomycin from biodegradable beads.

The current antibiotics delivery system for orthopedic infection treatment uses polymethylmethacrylate (PMMA) beads as a drug release. However the nonbiodegradable nature of the PMMA necessitates a second operation to remove the beads. This article explores the alternative of using biodegradable polymers as antibiotic beads for a long-term drug release. The effect of different processing factors on the release rate of the beads was investigated. To manufacture an antibiotic bead, polylactide-polyglycolide copolymers were mixed with vancomycin. The mixture was compressed and sintered at 55 degrees C to form beads of different sizes. An elution method was employed to characterize the release rate of antibiotic over a 35-day period at 37 degrees C. Biodegradable beads released high concentrations of antibiotic (well above the breakpoint sensitivity concentration) in vitro for the period of time needed to treat bone infection; i.e., 4-6 weeks. A bacterial inhibition test was carried out to determine the relative activity of the released antibiotics. The diameter of the sample inhibition zone ranged from 6.5-10 mm, which is equivalent to 12.5-100% of relative activity. By changing the processing parameters, we were able to control the release rate of the beads. This provides advantages of meeting the specific antibiotics requirement for patients with various surgical infections.

Anti-Bacterial Agents↗

PubMed1999

The value of analytical assays that are stability-indicating.

It is essential to validate an assay with regards to its precision, accuracy, reproducibility, selectivity and robustness. In pharmaceutical research, in addition to these requirements, an assay is often required to be proven beyond doubt to be stability-indicating. A stability-indicating assay is one that can accurately and selectively differentiate an intact drug from its potential decomposition products. An assay that fails to meet this requirement would produce stability data that are inaccurate and misleading. This concept could be applied to other fields of research, such as clinical laboratory chemistry where accurate and reliable clinical data are imperative in the diagnosis and following-up of diseases. In this study, we employed an amperometric HPLC assay developed for the measurements of urinary catecholamines and metanephrines to illustrate the validation process of this assay with regards to its stability-indicating capacity. The implications of this study are also discussed.

Catecholamines↗

PubMed1999

Factors affecting the formation of spherical bodies in the spirochete Treponema denticola.

The oral spirochete Treponema denticola typically is a helically shaped, motile bacterial cell. However, morphological variations of T. denticola cells in the form of "spherical bodies" are sometimes seen. Little is known about the environmental factors that cause their formation. The effects of oxygen, growth temperature, nutrient depletion and the addition of metabolic end-products were tested to determine their role in the morphogenesis of the spherical bodies. It was found that the age of the culture, the omission of individual components (yeast extract, rabbit serum, volatile fatty acids or thiamine pyrophosphate) from the medium and the addition of the metabolic end product lactic acid enhanced the formation of these bodies. However, their formation was decreased upon omission of the medium components asparagine and sodium bicarbonate.

Adaptation, Physiological↗