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Biomedical subjects

W Wei

Publications and source records attributed to W Wei.

At least 55 records · Page 3Linked to original sources
PubMed2001

Real-time monitoring of formaldehyde-induced DNA-lysozyme cross-linking with piezoelectric quartz crystal impedance analysis.

A novel method for monitoring, in real time, the formaldehyde (FA)-induced DNA-protein cross-linking process with the piezoelectric quartz crystal impedance (PQCI) technique is proposed. The method was used to monitor FA-induced DNA-lysozyme cross-link formation. Lysozyme was directly immobilized on the silver electrode surface of a piezoelectric quartz crystal by adsorption. The lysozyme-coated piezoelectric sensor was in contact with FA and DNA solutions. The time courses of the resonant frequency and equivalent circuit parameters of the sensor during the cross-linking were simultaneously obtained and are discussed in detail. On the basis of the feature of the multi-dimensional information provided by the PQCI technique, it was concluded that the observed frequency decrease could be mainly ascribed to the mass increase resulting from the cross-linking. According to the frequency decrease with time, the kinetics of the cross-linking process were quantitatively studied. A piezoelectric response model for the cross-linking was theoretically derived. Fitting the experimental data to the model, the kinetic parameters, such as the binding and dissociation rate constants (k(1) and k(-1)) and the cross-linking equilibrium constant (Ka), were determined. At 37 degrees C, the k(1), k(-1) and Ka values obtained were 7.0 (+/-0.1) x 10(-5) (microg ml(-1))(-1) s(-1), 6.6 (+/-0.1) x 10(-3) s(-1) and 1.06 (+/-0.02) x 10(-2) (microg ml(-1))(-1), respectively.

DNA↗
PubMed2001

Mutational analysis of the structure and functions of hepatitis C virus RNA-dependent RNA polymerase.

The hepatitis C virus (HCV) nonstructural protein 5B (NS5B) is an RNA-dependent RNA polymerase (RdRP), a central catalytic enzyme for HCV replication. To further understand the structure and functions of NS5B, we introduced a series of 27 clustered and 19 point substitution mutations within and outside the well-known motifs conserved among RdRP by alanine scanning and investigated effects of these mutants on enzymatic activity of NS5B. Surprisingly, most of the mutations (22 of 27 clustered mutants) do not affect RdRP activity at all, indicating that the side chains of the corresponding amino acid residues are dispensable for the catalytic activity. On the other hand, 4 mutants, cm20t, cm194t, cm2t, and cm3t, are defective in RdRP activity. By further analysis with point mutations within these regions, E18, Y191, C274, Y276, and H502 were determined to be critical for the RdRP activity. Y276 was also shown to be critical for RNA template/primer association, although 3 amino acid sequences were identified to be important for RNA template binding by RNA-filter binding assays. Finally, 4 discontinuous sequences of NS5B (aa139-145, aa149-155, aa 365-371, and aa 382-388) were found to be essential for binding to NS5A as determined by glutathione S-transferase (GST)-pull down assays using GST-NS5A and FLAG-NS5B expressed in cotransfected cells, and GST-pull down assay in vitro. In light of the crystal structure models of NS5B recently reported, our results indicate that the RdRP activity of NS5B requires the longer loop and the helix located at the distal of the thumb, which are unique among RdRPs as well as reverse transcriptases.

Amino Acid Sequence↗
PubMed2001

Is equity being sacrificed? Willingness and ability to pay for schistosomiasis control in China.

Decentralization of the health care system in China has led to an increasing need for income generation at all operational levels, both for curative services and for public health programmes. In general, people have accepted the costs of curative services, although the impact of charges on health-seeking behaviour has yet to be assessed. Public health programmes present particular problems in terms of revenue generation, however, because of the less direct impact of these activities on individual health and well-being. In this paper, we report the results of a cross-sectional study of willingness and ability to pay for schistosomiasis control. Questionnaires were administered to household heads of six representative villages in the Dongting Lake Region of Hunan Province, China. A total of 628 valid questionnaires were analyzed. The yearly mean household expenditure on schistosomiasis diagnosis and treatment was RMB 59.50 +/- 146.04 Yuan (US$1 = 8 RMB Yuan), accounting for 0.94% of the total yearly household income. Most household heads (514, 82%) thought schistosomiasis was the greatest health threat in their communities, but only 30.9% of them were willing to pay for screening, diagnosis and treatment of the infection. On the other hand, 72.3% of the respondents were willing to undertake volunteer work for control.

Adult↗
PubMed2001

Variation between two near isogenic barley (Hordeum vulgare) cultivars in expression of the B subunit of the vacuolar ATPase in response to salinity.

A gene encoding the barley vacuolar ATPase subunit B (BSVAP) was differentially expressed between two near isogenic barley cultivars, Golden Promise and Maythorpe. This gene (BSVAP) was isolated by the mRNA differential display technique (DDRT-PCR). BSVAP was salt inducible under long-term salinity stress in the salt sensitive cultivar Maythorpe but less so in the relatively salt tolerant Golden Promise and was more highly expressed under control conditions in Maythorpe. The physiological consequences of altered vacuolar ATPase expression are discussed in relation to the salt sensitivity of Maythorpe.

Adenosine Triphosphatases↗
PubMed2001

Role of p14(ARF) in replicative and induced senescence of human fibroblasts.

Following a proliferative phase of variable duration, most normal somatic cells enter a growth arrest state known as replicative senescence. In addition to telomere shortening, a variety of environmental insults and signaling imbalances can elicit phenotypes closely resembling senescence. We used p53(-/-) and p21(-/-) human fibroblast cell strains constructed by gene targeting to investigate the involvement of the Arf-Mdm2-p53-p21 pathway in natural as well as premature senescence states. We propose that in cell types that upregulate p21 during replicative exhaustion, such as normal human fibroblasts, p53, p21, and Rb act sequentially and constitute the major pathway for establishing growth arrest and that the telomere-initiated signal enters this pathway at the level of p53. Our results also revealed a number of significant differences between human and rodent fibroblasts in the regulation of senescence pathways.

Aging↗
PubMed2001

The gypsy insulator can act as a promoter-specific transcriptional stimulator.

Insulators define chromosomal domains such that an enhancer in one domain cannot activate a promoter in a different domain. We show that the Drosophila gypsy insulator behaves as a cis-stimulatory element in the larval fat body. Transcriptional stimulation by the insulator is distance dependent, as expected for a promoter element as opposed to an enhancer. Stimulation of a test alcohol dehydrogenase promoter requires a binding site for a GATA transcription factor, suggesting that the insulator may be facilitating access of this DNA binding protein to the promoter. Short-range stimulation requires both the Suppressor of Hairy-wing protein and the Mod(mdg4)-62.7 protein encoded by the trithorax group gene mod(mdg4). In the absence of interaction with Mod(mdg4)-62.7, the insulator is converted into a short-range transcriptional repressor but retains some cis-stimulatory activity over longer distances. These results indicate that insulator and promoter sequences share important characteristics and are not entirely distinct. We propose that the gypsy insulator can function as a promoter element and may be analogous to promoter-proximal regulatory modules that integrate input from multiple distal enhancer sequences.

Alcohol Dehydrogenase↗
PubMed2001

Human L1 retrotransposition: cis preference versus trans complementation.

Long interspersed nuclear elements (LINEs or L1s) comprise approximately 17% of human DNA; however, only about 60 of the approximately 400,000 L1s are mobile. Using a retrotransposition assay in cultured human cells, we demonstrate that L1-encoded proteins predominantly mobilize the RNA that encodes them. At much lower levels, L1-encoded proteins can act in trans to promote retrotransposition of mutant L1s and other cellular mRNAs, creating processed pseudogenes. Mutant L1 RNAs are mobilized at 0.2 to 0.9% of the retrotransposition frequency of wild-type L1s, whereas cellular RNAs are mobilized at much lower frequencies (ca. 0.01 to 0.05% of wild-type levels). Thus, we conclude that L1-encoded proteins demonstrate a profound cis preference for their encoding RNA. This mechanism could enable L1 to remain retrotransposition competent in the presence of the overwhelming number of nonfunctional L1s present in human DNA.

Base Sequence↗
PubMed2001

Association of smoking with serum and dietary levels of antioxidants in adults: NHANES III, 1988-1994.

OBJECTIVES: This study examined the association of smoking with serum levels and dietary intakes of antioxidants in a nationally representative sample. METHODS: This study classified 7873 apparently healthy adults aged 17 to 50 years from National Health and Nutrition Examination Survey III (NHANES III) data as nonsmokers or as smokers if their serum cotinine levels were either lower than 14 ng/mL or 14 ng/mL or greater, respectively. SUDAAN software was used for the statistical analysis. RESULTS: Smokers of both sexes had significantly (P < .001) lower serum levels of vitamin C, alpha-carotene, beta-carotene, beta-cryptoxanthin, and lutein/zeaxanthin. Reduction in the serum vitamin E, lycopene, and selenium levels in smokers was slight. Smokers also had significantly lower dietary intakes of vitamin C and beta-carotene. A significant (P < .001) inverse relation was found between serum vitamin C and beta-carotene levels and cotinine levels independent of diet effect, and a positive relation (P < .001) was found between serum levels and dietary intakes. CONCLUSIONS: Antioxidants appear to have differing declines in serum levels as a result of reduced dietary intakes and the effects of smoking.

Adolescent↗
PubMed2001

Improvement of melatonin to the learning and memory impairment induced by amyloid beta-peptide 25 - 35 in elder rats.

AIM: To investigate improvement of melatonin on learning and memory impairment induced by amyloid beta-peptide 25 - 35 (Abeta25-35) in elder rats. METHODS: Step-down type passive avoidance test, shuttle-box test, and Morris water maze were used together to determine effects of Abeta25-35 and melatonin on learning and memory. Pathological changes were observed by HE, Congo red, and Ag staining. RESULTS: The elder rats were injected bilaterally Abeta25-35 20 microg into the hippocampus to induce learning and memory dysfunction. Melatonin administration (0.1, 1, and 10 mg/kg, ig x 8 d) to the Abeta25-35-treated rats prolonged the latency, shortened the total stimulating time, and decreased the number of errors in the step-down test. Shuttle-box test showed that melatonin improved amnesic rats' performance at the same doses. Melatonin (0.1, 1, and 10 mg/kg ), giving for 10 d, could enhance the spatial resolution of amnesic rats in Morris water maze test. Also in Abeta25-35-treated group, a decrease in the number of neurons in cortex and hippocampus, a massive glial reaction, and neurophilic phenomenon were detected by HE staining; the positive vascular amyloidosis by Congo red and fibrils by Ag staining were observed. Melatonin (0.1 and 1 mg/kg)could inhibit above pathological changes in Abeta25-35 group. CONCLUSION: Melatonin improved the impaired learning and memory induced by Abeta25-35 in elder rats.

Amyloid beta-Peptides↗
PubMed2001

Balanced ultrafiltration, modified ultrafiltration, and balanced ultrafiltration with modified ultrafiltration in pediatric cardiopulmonary bypass.

This study evaluates the effect of balanced ultrafiltration, modified ultrafiltration, and balanced ultrafiltration with modified ultrafiltration on inflammatory mediators in children's open-heart surgery. Eighty children with congenital heart disease were randomly divided into four groups: control group (C group); balanced ultrafiltration group (BUF group); modified ultrafiltration group (MUF group); and balanced ultrafiltration with modified ultrafiltration group (B+M group). Clinical data of these groups were similar. Tumor necrosis factor (TNF), interleukin-8(IL-8), and E-selectin were measured at the beginning of cardiopulmonary bypass (CPB), 30 min later, at the cessation of CPB, at the cessation of MUF (MUF group and B+M group), and 2 hours postoperatively. During CPB, the concentrations of TNF, IL-8, and E-selectin increased significantly in C and MUF groups and did not change significantly in BUF and B+M groups. In the period of MUF, TNF and IL-8 increased; whereas, E-selectin did not change. The study shows that ultrafiltration can filter out the inflammatory mediators, but only BUF can decrease the concentrations of them. Moreover, MUF only can concentrate blood. Combining both techniques has both effects, but the effect of BUF was offset by MUF.

Cardiopulmonary Bypass↗
PubMed2001

[The morphologic structure of vitreous hemorrhage and posterior vitreous detachment].

OBJECTIVE: To investigate the morphologic structures of vitreous hemorrhage that may be beneficial to its treatment. METHODS: The clinical data of 74 cases (79 eyes) with vitreous hemorrhage resulting from proliferative diabetic retinopathy, retinal vasculitis and retinal vein occlusion in December 1995 to January 2000 and having undergone vitrectomy were studied retrospectively, which focused on the morphologic characteristics of vitreous body by B-scan, surgical microscopy and surgical manipulation, to evaluate the relation of retina and vitreous body in order to discern the relationships among the retina, neovascular fibromembrane and the posterior hyaloid surface. RESULTS: Posterior vitreous detachment (PVD) in various degrees was observed in all the cases. The morphologic character was classified as the complete and partial vitreous detachment. The latter one can be subdivided into "V", "L" types and posterior vitreoschisis that was not rarely seen in the proliferative phase of ischemic retinopathy. CONCLUSIONS: To evaluate the characteristics of posterior vitreous detachment and vitreoschisis may provide valuable information for vitreoretinal surgery, elevate the rate of success and improve the prognosis of the surgery.

Adult↗
PubMed2001

[Diagnosis and surgical treatment of carotid body tumor].

OBJECTIVE: To promote the rate of diagnosis and successful rate of operation for carotid body tumor. METHOD: 26 cases of the 30 patients with carotid body tumor were performed by DSA and 5 cases of them, feeding arteries of tumor were embolized with gelfoam through superlective catheterization, preoperativelly. The tumor was resected under carotid adventitial plane in 12 cases, with extracervical artery in 8 cases and with tumor-carotid segment block in 10 cases. RESULT: Of the 30 cases, no one died from operation or cerebral vascular complications. DSA plays a significant role in diagnosis of the tumor. Intraoperative bleeding can be remarkably reduced if feeding arteries were embolized preoperativelly. The tumor fixed to segment of the carotid arteries block was resected and the reconstruction of vasculature was unnecessary as soon as candidates were certified to be qualified by carotid training and DSA or/and TCD tests. Follow up of these 30 patients revealed that 25 of them survived free from cerebrovascular complications and tumor recurrence for 5 months-10 years. 1 of them died from traffic accident 2 year after operation, 4 of them had lost communication. CONCLUSION: DSA is a most authritative approach for diagnosis of carotid body tumor, DSA and TCD tests play a key role to resect carotid artery without reconstruction. Successful operation based on a precise preoperative evaluation, choice of operative ways and proper procedure.

Adult↗
PubMed2000

Polarity of transcriptional enhancement revealed by an insulator element.

Transcriptional enhancers for genes transcribed by RNA polymerase II may be localized upstream or downstream of the stimulated promoter in their normal chromosomal context. They stimulate transcription in an orientation-independent manner when assayed on circular plasmids. We describe a transient transformation system to evaluate the orientation preference of transcriptional enhancers in Drosophila. To accomplish this, the gypsy insulator element was used to block bidirectional action of an enhancer on circular plasmids. In this system, as in the chromosome, blocking of enhancer activity requires wild-type levels of the su(Hw) protein. We evaluated the orientation preference for the relatively large (4.4 kb) Adh larval enhancer from Drosophila melanogaster, used in conjunction with a luciferase reporter gene under the control of a minimal Adh promoter. An orientation preference was revealed by insertion of a single copy of the insulator between the enhancer and the promoter. This orientation effect was greatly amplified when the promoter was weakened by removing binding sites for critical transcription factors, consistent with a mechanism of insulator action in which the insulator intercepts signals from the enhancer by competing with the promoter. The orientation preference, as much as 100-fold, is a property of the enhancer itself because it is displayed by gene constructions introduced into the chromosome regardless of the presence of the insulator in a distal location. These findings are most easily reconciled with a facilitated tracking mechanism for enhancer function in a native chromosomal environment.

Animals↗
PubMed2000

Functional interaction between p53 and the interferon-inducible nucleoprotein IFI 16.

Interferons are important in regulating cell growth and differentiation, immune function and initiating anti-viral responses. While the pleotrophic actions of interferons have been well documented, the molecular mechanisms underpinning their biological effects have not been fully characterized. IFI 16 is a member of the interferon-inducible HIN-200 family of nuclear proteins, which we have recently shown can function as a potent transcriptional repressor. A murine member of the HIN-200 family, p202, can indirectly interact with p53 via the p53 binding protein (p53bp) and inhibit p53-mediated transcriptional activation. The binding activity of p202 to p53bp was shown to require the conserved MFHATVAT motif present in all 200 amino acid repeat regions of HIN-200 proteins. Given that IFI 16 contains two MFHATVAT motifs, we sought to determine whether IFI 16 may form a complex with p53 and if so to ascertain the functional significance of this interaction. We demonstrate that IFI 16 can directly bind to the C-terminal region of p53 and augment p53-mediated transcriptional activation without altering the steady state levels of p53. Thus, in addition to its ability to directly regulate gene expression, IFI 16 can also modulate the transcription function of other cellular transcription factors. These findings demonstrate a possible link between gene induction following interferon stimulation and p53-mediated cellular events.

Amino Acid Motifs↗
PubMed2000

Determination of divalent carboxylic acids in traditional Chinese herbal and patent medicines using HPLC with double cell quartz crystal detector.

A rapid, simple and sensitive high performance liquid chromatography (HPLC) method combined with a novel double cell quartz crystal (DCQC) detector is described for the determination of divalent carboxylic acids in traditional Chinese herbal and patent medicines. The chromatographic system involves the use of a muBondapak-NH2 column with 20 mM KH2PO4 (pH 3.4, adjusted by H3PO4)/acetonitrile (95:5%) as a mobile phase. The DCQC detector has a low temperature coefficient and high conductance sensitivity independent of the background from 7.2 to 2500 microS. Analysis of sample is completed without the use of an ion suppression device.

Calibration↗
PubMed2000

Improvements in DNA sequencing by capillary electrophoresis at elevated temperature using poly(ethylene oxide) as a sieving matrix.

DNA sequencing in poly(ethylene oxide) (PEO) matrix by capillary electrophoresis was demonstrated at high temperature. The optimal separation temperature is around 40 degrees C. The effects of polymer concentration and types of buffers on the separation performance were investigated. A new buffer system consisting of Tris-Taps-His-EDTA works well with PEO. High-speed separation and good resolution can be fulfilled by using a single-MW PEO polymer. It offers similar separation performance as before for the small DNA fragments, but better performance for large DNA fragments.

Buffers↗
PubMed2000

alpha-Amylase immobilized on bulk acoustic-wave sensor by UV-curing coating.

A new method for immobilization of alpha-amylase by UV-curing coating is proposed in this paper. The immobilization procedure of UV-curing coating on piezoelectric quartz crystal is simple and convenient, and causes less loss of enzymatic activity. The activity of the immobilized alpha-amylase is monitored by a technique based on bulk acoustic-wave (BAW) sensor. The frequency shift of BAW sensor can reflect the degree of hydrolysis of starch by the immobilized alpha-amylase. It is appropriate for the immobilized alpha-amylase to hydrolyze the soluble starch under pH 7.0 condition, which is similar to that of the free alpha-amylase. Kinetic parameters (the Michaelis constant, K(m), and the maximum initial rate V(max)) of the enzymatic hydrolysis of starch by the immobilized alpha-amylase are estimated by using a linear method of Lineweaver-Burk plot. K(m)=12.7mgml(-1) and V(max)=15.9Hzmin(-1). And the experimental results show that the immobilized alpha-amylase entrapped by the UV-curing coating retains adequate enzymatic activity and can be reused more than 50 times under certain experimental conditions.

Journal Article↗