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Biomedical subjects

S Q Wang

Publications and source records attributed to S Q Wang.

At least 19 recordsLinked to original sources
PubMed2006

Antisense oligonucleotides targeted against asialoglycoprotein receptor 1 block human hepatitis B virus replication.

Chronic hepatitis B virus (HBV) infection is a major worldwide public health problem. Better therapeutics and treatment strategies are urgently needed because of ineffective clinical treatment. Our previous study showed that asialoglycoprotein receptor 1 (ASGPR1) was upregulated by HBV but downregulated by lamivudine in HepG2.2.15 cells. It has also been reported that ASGPR is a candidate receptor for HBV attachment to hepatocytes. Therefore, as a major subunit of ASGPR, ASGPR1, might be a potential target for anti-HBV drugs. To validate this hypothesis, antisense oligonucleiotides (ASODNs) were used to downregulate ASGPR1 level in HepG2.2.15 cells. By using the MFOLD web server and BLAST searches, five ASODNs theoretically targeting ASGPR1 were selected. After 72 h post-transfection, HBV-DNA level in cell medium were examined by real-time polymerase chain reaction (PCR). Hepatitis B surface antigen (HBsAg) and Hepatitis B e antigen (HBeAg) were detected using enzyme-linked immunosorbent assay (ELISA). ASGPR1 mRNA and protein level were measured by semi-quantitative reverse transcriptase (RT)-PCR and Western blot analysis respectively. The results showed that ASODN2 significantly downregulated ASGPR1 level. It also reduced HBV-DNA, HBsAg and HBeAg level in cell medium as observed with lamivudine. In contrast, the sense sequence and scrambled sequence of ASODN2 had no effect on ASGPR1 and HBV markers in HepG2.2.15 cells. This indicated that ASODN2 could specifically reduce HBV replication in vitro. Additionally, cell proliferation and apoptosis assay suggested that downregulation of ASGPR1 did not affect cell viability. We, therefore, proposed that ASODNs targeted against ASGPR1 could block HBV replication without the influence of other changes, and ASGPR1 could be targeted for anti-HBV drug development.

Antiviral Agents↗
PubMed2005

HBx protein of hepatitis B virus (HBV) can form complex with mitochondrial HSP60 and HSP70.

HBx, a transcriptional transactivating protein of hepatitis B virus (HBV), is required for viral infection and has been implicated in virus-mediated liver oncogenesis. However, the molecular mechanism for its influence on cell remains largely unknown. It was proved that HBx need the help of host cell proteins to exert its function by binding to them. During purifying of GSTX (fusion protein of GST and HBx) expressed in E. coli, we found that it can bind specifically with GrpE (HSP60) and DnaK (HSP70) of E. coli while GST cannot. Using GST pull-down, two-dimensional gel electrophoresis and mass spectrum, we found that GSTX can also bind to human mitochondrial HSP60 and HSP70, which are homologues of GrpE and DnaK. These interactions between HBx and mitochondrial HSP60 and HSP70 are supported by the result of co-immunoprecipitation experiment. It means that HBx can form complex with E. coli and human HSP60 and HSP70. The implication of HBx, HSP60 and HSP70 complex in molecular mechanism of virus infection is discussed.

Amino Acid Sequence↗
PubMed2005

Quantification of the relative levels of wild-type and lamivudine-resistant mutant virus in serum of HBV-infected patients using microarray.

During the course of lamivudine administration in hepatitis B virus (HBV)-infected patients, a dynamic development of the viral population in serum is observed. Total HBV level is initially reduced, then lamivudine-resistant mutants appear, and finally, the viral level is increased. All methods of mutant detection so far described can only identify mutants in the serum, and cannot determine the proportion of those mutants. In this paper, we report the development of a novel technique that can quantify the relative proportion of mutants in serum utilizing gene microarray technology. Based on the nucleotide sequence at the loci of the mutations in lamivudine-resistant HBV mutants, 28 probes were designed and dotted on glass film to prepare the oligonucleotide microarray. Ten standard curves were established by employing 15 reference plasmids as templates. Ten standard math functions were simulated, which allowed quantification of the proportion of mutants in the sample by measuring the value of fluorescent intensity on the microarray. By utilizing the standard math function, the relative proportion of two different mutation sequences in the mixed template could be detected with an error <10%. The HBV-lamivudine oligonucleotide microarray is reliable to quantify the relative proportion of wild-type HBV vs HBV mutants in patient's sera.

Base Sequence↗
PubMed2005

Differentially expressed cellular genes following HBV: potential targets of anti-HBV drugs?

The aim of the study was to screen for cellular genes that are differentially expressed following hepatitis B virus (HBV) infection, in an attempt to identify potential targets of anti-HBV drugs. An oligonucleotide microarray containing 231 virus-infection-associated genes was prepared. Differential gene expression in HepG2.2.15 cells compared to control with HepG2 cells was analysed by this in-house microarray. The change in gene expression in HepG2.2.15 cells treated by lamivudine on days 4 and 8 after exposure was also studied. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to comfirm the differentially expressed genes induced by HBV and lamivudine. There were 31 upregulated and four downregulated genes in HepG2.2.15 cells compared with the HepG2 control cells. Eleven genes were consistently altered by lamivudine at both time points. Of the 31 genes that were upregulated in HepG2.2.15 cells, there were seven genes which were downregulated by lamivudine. Of the four downregulated genes, there was one gene which was upregulated by lamivudine. Of the differentially expressed genes induced by HBV and lamivudine, the expression of five genes was confirmed by semi-quantitative RT-PCR. These results shed new light on the effects of HBV and lamivudine on cellular gene expression. Differentially expressed genes induced by HBV and lamivudine could potentially become new anti-HBV drug targets in novel therapies.

Antiviral Agents↗
PubMed2005

Oligonucleotide microarray for HLA-DRB1 genotyping: preparation and clinical evaluation.

In this study, HLA-DRB1 gene was genotyped by using the microarray technique. Oligonucleotide probes were designed based on partial sequences of various genotypes of HLA-DRB1, and were fixed on a silylated slide to form a microarray. The second exon of HLA-DRB1 gene in the extracted genomic DNA samples was amplified and labelled by means of polymerase chain reaction (PCR); then it was hybridized to the microarray. The microarray was scanned, and the result was analysed in order to determine the genotypes of HLA-DRB1 of the tested sample. A total of 1574 of 1592 clinical samples had accordant results of genotypes in either microarray assay or PCR-SSP assay; 8 of 10 samples that had inconsistent results of genotypes were proved to be microarray-assay reliable by confirmation of DNA sequencing. It is concluded that microarray is an alternative reliable method for HLA-DRB1 genotyping.

China↗
PubMed2001

Site-specific antibody of (Na(+) + K(+))-ATPase augments cardiac myocyte contraction without inactivating enzyme activity.

(Na(+) + K(+))-ATPase regulates both excitability and contractility of the heart. Little is known about the molecular basis of the enzyme that underlies its cardiac regulatory functions. Here we demonstrate that the (833)KRQPRNPKTDKLVNE(847) region, which resides in the alpha-subunit of rat (Na(+) + K(+))-ATPase, directly participates in the regulation of cardiac contraction. A site-specific antibody (SSA95) against this peptide sequence markedly increased intracellular Ca(2+) transients and contraction (EC(50) = 11.4 nM) in intact rat heart cells without inactivating the (Na(+) + K(+))-ATPase. These novel findings establish the first link between a precise structural region of the (Na(+) + K(+))-ATPase and cardiac positive inotropy.

Amino Acid Sequence↗
PubMed2001

FTIR images.

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Journal Article↗
PubMed2001

beta-Adrenergic stimulation synchronizes intracellular Ca(2+) release during excitation-contraction coupling in cardiac myocytes.

To elucidate microscopic mechanisms underlying the modulation of cardiac excitation-contraction (EC) coupling by beta-adrenergic receptor (beta-AR) stimulation, we examined local Ca(2+) release function, ie, Ca(2+) spikes at individual transverse tubule-sarcoplasmic reticulum (T-SR) junctions, using confocal microscopy and our recently developed technique for release flux measurement. beta-AR stimulation by norepinephrine plus an alpha(1)-adrenergic blocker, prazosin, increased the amplitude of SR Ca(2+) release flux (J(SR)), its running integral (integralJ(SR)), and L-type Ca(2+) channel current (I(Ca)), and it shifted their bell-shaped voltage dependence leftward by approximately 10 mV, with the relative effects ranking I(Ca)> J(SR)>integralJ(SR). Confocal imaging revealed that the bell-shaped voltage dependence of SR Ca(2+) release is attributable to a graded recruitment of T-SR junctions as well as to changes in Ca(2+) spike amplitudes. beta-AR stimulation increased the fractional T-SR junctions that fired Ca(2+) spikes and augmented Ca(2+) spike amplitudes, without altering the SR Ca(2+) load, suggesting that more release units were activated synchronously among and within T-SR junctions. Moreover, beta-AR stimulation decreased the latency and temporal dispersion of Ca(2+) spike occurrence at a given voltage, delivering most of the Ca(2+) at the onset of depolarization rather than spreading it out throughout depolarization. Because the synchrony of Ca(2+) spikes affects Ca(2+) delivery per unit of time to contractile myofilaments, and because the myofilaments display a steep Ca(2+) dependence, our data suggest that synchronization of SR Ca(2+) release represents a heretofore unappreciated mechanism of beta-AR modulation of cardiac inotropy.

Action Potentials↗
PubMed2001

Ca2+ signalling between single L-type Ca2+ channels and ryanodine receptors in heart cells.

Ca2+-induced Ca2+ release is a general mechanism that most cells use to amplify Ca2+ signals. In heart cells, this mechanism is operated between voltage-gated L-type Ca2+ channels (LCCs) in the plasma membrane and Ca2+ release channels, commonly known as ryanodine receptors, in the sarcoplasmic reticulum. The Ca2+ influx through LCCs traverses a cleft of roughly 12 nm formed by the cell surface and the sarcoplasmic reticulum membrane, and activates adjacent ryanodine receptors to release Ca2+ in the form of Ca2+ sparks. Here we determine the kinetics, fidelity and stoichiometry of coupling between LCCs and ryanodine receptors. We show that the local Ca2+ signal produced by a single opening of an LCC, named a 'Ca2+ sparklet', can trigger about 4-6 ryanodine receptors to generate a Ca2+ spark. The coupling between LCCs and ryanodine receptors is stochastic, as judged by the exponential distribution of the coupling latency. The fraction of sparklets that successfully triggers a spark is less than unity and declines in a use-dependent manner. This optical analysis of single-channel communication affords a powerful means for elucidating Ca2+-signalling mechanisms at the molecular level.

Animals↗
PubMed2001

Typical dermoscopic patterns of benign melanocytic nevi.

Dermoscopy often is an effective diagnostic tool for differentiating benign and malignant pigmented skin lesions. The observed dermoscopic structures and patterns of lesions provide valuable sources of information that are not accessible by routine clinical examination. Recognition and interpretation of these dermoscopic structures and patterns require training and experience. Mentioned here are some fundamental guidelines for differentiating malignant and benign lesions. Some of the classic patterns associated with a list of benign melanocytic nevi also have been illustrated. Additional research is needed to identify other patterns of these benign lesions. More importantly, analysis of the prevalence of various patterns associated with the various benign lesions will be helpful to clinicians having the responsibility of making in vivo diagnoses of benign compared with malignant melanocytic neoplasms.

Dermatology↗
PubMed2001

Reduction of ultraviolet transmission through cotton T-shirt fabrics with low ultraviolet protection by various laundering methods and dyeing: clinical implications.

BACKGROUND: The public has long been instructed to wear protective clothing against ultraviolet (UV) damage. OBJECTIVE: Our purpose was to determine the UV protection factor (UPF) of two cotton fabrics used in the manufacture of summer T-shirts and to explore methods that could improve the UPF of these fabrics. METHODS: Each of the two types of white cotton fabrics (cotton T-shirt and mercerized cotton print cloth) used in this study was divided into 4 treatment groups: (1) water-only (machine washed with water), (2) detergent-only (washed with detergent), (3) detergent-UV absorber (washed with detergent and a UV absorber), and (4) dyes (dyed fabrics). Ultraviolet transmission through the fabrics was measured with a spectrophotometer before and after laundry and dyeing treatments. Based on UV transmission through these fabrics, the UPF values were calculated. RESULTS: Before any treatments, the mean UPFs were 4.94 for the T-shirt fabric and 3.13 for the print cloth. There was greater UVA (320-400 nm) than UVB (280-320 nm) transmission through these fabrics. After 5 washings with water alone and with detergent alone, UPF increased by 51% and 17%, respectively, for the cotton T-shirt fabric. Washing the T-shirt fabrics with detergent plus the UV-absorbing agent increased the UPF by 407% after 5 treatments. Dyeing the fabric blue or yellow increased the UPF by 544% and 212%, respectively. Similar changes in UPFs were observed for the print cloth fabric. CONCLUSION: The two cotton fabrics used in this study offered limited protection against UV radiation as determined by spectrophotometric analysis. Laundering with detergent and water improves UPF slightly by causing fabric shrinkage. Dyeing fabrics or adding a UV-absorbing agent during laundering substantially reduces UV transmission and increases UPF. More UVA is transmitted through the fabrics than UVB.

Coloring Agents↗
PubMed2001

Ultraviolet A and melanoma: a review.

The incidence and mortality rates of melanoma have risen for many decades in the United States. Increased exposure to ultraviolet (UV) radiation is generally considered to be responsible. Sunburns, a measure of excess sun exposure, have been identified as a risk factor for the development of melanoma. Because sunburns are primarily due to UVB (280-320 nm) radiation, UVB has been implicated as a potential contributing factor to the pathogenesis of melanoma. The adverse role of UVA (320-400 nm) in this regard is less well studied, and currently there is a great deal of controversy regarding the relationship between UVA exposure and the development of melanoma. This article reviews evidence in the English-language literature that surrounds the controversy concerning a possible role for UVA in the origin of melanoma. Our search found that UVA causes DNA damage via photosensitized reactions that result in the production of oxygen radical species. UVA can induce mutations in various cultured cell lines. Furthermore, in two animal models, the hybrid Xiphophorus fish and the opossum (Mondelphis domestica), melanomas and melanoma precursors can be induced with UVA. UVA radiation has been reported to produce immunosuppression in laboratory animals and in humans. Some epidemiologic studies have reported an increase in melanomas in users of sunbeds and sunscreens and in patients exposed to psoralen and UVA (PUVA) therapy. There is basic scientific evidence of the harmful effects of UVA on DNA, cells and animals. Collectively, these data suggest a potential role for UVA in the pathogenesis of melanoma. To date evidence from epidemiologic studies and clinical observations are inconclusive but seem to be consistent with this hypothesis. Additional research on the possible role of UVA in the pathogenesis of melanoma is required.

Animals↗
PubMed2001

[Construction of a deep coverage rice BAC library and identification of clones associated with disease-resistant genes].

A BAC library for IRBB56, an accession pyramiding Xa4, xa5 and xa13 three bacterial blight resistance genes, was constructed. The library contains 55,296 clones with an average insert size of 132 kb. Based on a haploid genome size of 450 Mb, the coverage of the library was about 14 genome equivalents that make it one of the most comprehensive BAC libraries available in rice and provide 99.99% possibility to isolate any interested rice genes or sequences in the library. To determine the representation of organelle DNA homologues in the library, the library was screened with three different chloroplast genes and four mitochondrial genes, respectively. Results from this screening showed that less than 1% of clones in the library contain organelle genomic DNA homologues. Then, DNA markers on three different chromosomes linked to Xa4, xa5, and xa13, and a PCR fragment of rice UROD gene, were used to screen the library resulting in a range of 11-106 hits that will promote the isolation of these genes. The deep coverage and the large insert size of the library will facilitate physical mapping, isolation, and cloning of rice genes.

Chromosomes, Artificial, Bacterial↗
PubMed2001

[The construction and evaluation of antisense RNA expression vector targeting at HCV 5'NCR].

Antisense RNA is an important field of antisense technique. In order to explore a novel approach for the treatment of hepatitis C virus (HCV) infection and prove the availability of transgenic cellular model HepG2. 9706, an antisense RNA was designed targeting at the highly conserved 5'NCR and translation initiation site of HCV RNA at nt positions 13-397. It was inserted into the downstream of SV40 promotor sequence of pGL3 control vector in which luciferase gene is deleted. The antisense RNA expression vector (pHCV-asR) was constructed and identified by PCR, endonucleases reaction and DNA sequencing. Its expression in transfected HepG2 cells was tested by RT-PCR. To evaluate the inhibitory activities of pHCV-asR, HepG2. 9706 cells were transfected using this construct via Lipofectin method. Luciferase activity in cell lysates was measured for quantitative determining antiviral effects within the cells. The results showed that the inserted sequence of the pHCV-asR is the same as the designed sequence and can express in HepG2 cells. It was also found that pHCV-asR in HepG2. 9706 has a dose-dependent inhibitory effects on luciferase expression controlled by HCV 5'NCR with a inhibitory rate of 57%.

5' Untranslated Regions↗
PubMed2001

Interaction of Cd and citric acid, EDTA in red soil.

Adsorption and desorption process of cadmium in red soil (Ferrisols) as well as the influence by media's pH were investigated in detail with and without citric acid and EDTA. Experimental results clearly showed that Cd adsorption in red soil was affected significantly by the coexisted organic chemicals. In the presence of citric acid and EDTA, Cd adsorption in red soil increased with pH in acid media but decreased in high pH one. Further studies placed stress on the adsorbed Cd in red soil which was found to be existed mainly as exchangeable one at pH < 5.5, and desorption rate by 0.10 mol/L NaNO3 gave a peak-shaped curve due to the difference of specifically and nonspecifically adsorbed Cd with pH's change.

Adsorption↗
PubMed2001

Induction of apoptosis by superoxide anion and the protective effects of selenium and Vitamin E.

OBJECTIVE: The purpose of this study is to investigate the effect of superoxide anion on the apoptosis of cultured fibroblasts and the protective role of selenium and Vitamin E. METHODS: Cultured fibroblasts (NIH3T3), with or without selenium or vitamin E in the medium, were treated by superoxide anion produced by xanthine/xanthine oxidase reaction system and changes in cell structure and DNA were observed microscopically and electrophoretically. RESULTS: Apoptosis was observed when superoxide anion at a concentration of 5 nmol/L or 10 nmol/L had acted on the fibroblasts for 5-10 h. Selenium and Vitamin E in the medium inhibited the apoptosis significantly when their concentrations reached 1.15 mol/L and 2.3 mol/L respectively. CONCLUSION: Selenium and vitamin E have protective effect against the apoptosis induced by superoxide anion. The effect of selenium is more remarkable than that of vitamin E.

Antioxidants↗
PubMed2001

[Preparation optimization and properties of the aldehyde microscopic slides for oligonucleotide microarray fabrication].

The process for preparing the aldehyde slides was optimized and the properties of the aldehyde microscopic slides for immobilizing oligonucleotide were explored. The result shows that the concentration of aminosliane reagent plays an important role in the fluorescent background. Aldehyde slides with 2% aminosilane and 5% aldehyde treatment for 16 min and 30 min respectively immobilize oligonucleotide efficiently and have low fluorescence background. During oligonucleotide immobilization, terminal amino modification has no obvious specificity, but it can enhance the hybridization capacity of immobilized oligonucleotides. At low concentration (less than 10 mumol/L), hybridization signal has linear relationship with probe concentration, the hybridization signal reaches saturation when probe concentration is more than 20 mumol/L.

Aldehydes↗