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Biomedical subjects

Wei Wang

Publications and source records attributed to Wei Wang.

At least 109 records · Page 6Linked to original sources

Sequence of mammalian fossils, including hominoid teeth, from the Bubing Basin caves, South China.

A Plio-Pleistocene to Holocene faunal sequence has been recovered from four carefully excavated caves in the Bubing Basin, adjacent to the larger Bose Basin of South China. The caves vary in elevation; we suggest that the higher caves were formed and filled with sediments prior to the lower caves. The highest deposits, which are from Mohui Cave, contain hominoid teeth and other fossilized remains of mammalian taxa most similar to late Pliocene and early Pleistocene faunas. Wuyun Cave ( approximately 50m lower in elevation than Mohui) contains a late middle Pleistocene fauna, which is supported by U-series age constraints from 350 to 200ka. Lower Pubu Cave ( approximately 23m below Wuyun) is assigned to the late Pleistocene, while the Cunkong Cave (the lowest, approximately 2m lower elevation than Lower Pubu) preserves a Holocene fauna. The four faunal assemblages indicate species-level changes in Ailuropoda, Stegodon, and Sus, the appearance of Elephas, the local disappearance of Stegodon, and the migration of Equus hemionus to South China. These initial results of our work call into question the continued value of the Stegodon/Ailuropoda Fauna, a category long used to characterize the Pleistocene faunas of South China. Excavation of karstic caves of varying elevation within the basins of South China holds promise for defining local sequences of mammalian fossils that can be used to investigate faunal variations related to climate change, biogeographic events, and evolutionary change over the past two million years. Stable isotopic analysis of a small sample of mammalian teeth from Bubing Basin caves is consistent with 100% C(3) vegetation in the Bubing/Bose region, with certain delta(13)C values consistent with a canopied woodland or forest. A preliminary assessment of the hominoid teeth indicates the presence of diverse molar and premolar morphologies including dental remains of Gigantopithecus blacki and a sample with similarities to the teeth reported from Longgupo.

Animals↗

A new method for quantifying residue conservation and its applications to the protein folding nucleus.

The conservation of residues in columns of a multiple sequence alignment (MSA) reflects the importance of these residues for maintaining the structure and function of a protein. To date, many scores have been suggested for quantifying residue conservation, but none has achieved the full rigor both in biology and statistics. In this paper, we present a new approach for measuring the evolutionary conservation at aligned positions. Our conservation measure is related to the logarithmic probabilities for aligned positions, and combines the physicochemical properties and the frequencies of amino acids. Such a measure is both biologically and statistically meaningful. For testing the relationship between an amino acid's evolutionary conservation and its role in the Phi-value defined protein folding kinetics, our results indicate that the folding nucleus residues may not be significantly more conserved than other residues by using the biological-relevance weighted statistical scoring method suggested in this paper as an alternative to entropy-based procedures.

Amino Acid Sequence↗

Hepatitis B virus X protein (HBx) activates ATF6 and IRE1-XBP1 pathways of unfolded protein response.

Numerous viruses including hepatitis B virus (HBV) induce endoplasmic reticulum (ER) stress, which interrupts protein folding causing accumulation of unfolded or misfolded proteins in ER. To alleviate the stress placed on ER, these proteins must be refolded or degraded by activating a specific cellular response known as ER stress response or unfolded protein response (UPR). Two UPR-specific signaling pathways involving transmembrane proteins ATF6 and XBP1 generate critical transcription factors that activate UPR-responsive genes. In this study, the role of the multifunctional regulatory protein of HBV (HBx protein) in activation of UPR was investigated. In Hep3B cells with transit or stable expression of HBx, XBP1 expression and ATF6 cleavage was observed, suggesting that the ATF6 and IRE1-XBP1 pathways were activated. Furthermore, these two pathways were also activated in HepG2.2.15 cells that constitutively replicate the intact HBV genome, and blocked at least partly by cotransfection with small interfering RNA (siRNA) expression plasmid that knocked down HBx expression. Our results clearly establish HBx as an inducer of UPR and the activator of the ATF6 and IRE1-XBP1 pathways of UPR. HBx-mediated activation of these pathways of UPR probably promote HBV replication and expression in liver cells, and contribute to liver pathogenesis, perhaps even to hepatocellular carcinoma (HCC) development.

Activating Transcription Factor 6↗

Clinical and serological study of myasthenia gravis in HuBei Province, China.

BACKGROUND: Ocular and childhood myasthenia gravis (MG) cases seem relatively more common in Oriental than in Caucasian populations, but there have been no comprehensive serological studies on patients from mainland China. METHODS: 391 unselected patients with MG attending Tongji Hospital in WuHan (the largest hospital in the province of HuBei, China) were studied during a 15-month period; most had already received treatment for their condition. RESULTS: The male to female ratio was 0.8. 50% of the patients were children (<15 years), and age at onset showed a single peak at between 5 and 10 years of age. 64% of the children and 66% of the adults were positive for acetylcholine receptor (AChR) antibodies but the antibody titres were lower than in similar Caucasian studies, although this was partly due to the high incidence of ocular MG. Of the 43 patients with generalised MG without AChR antibodies, only 1 had muscle-specific kinase antibodies (2.5%) and 2 had voltage-gated calcium channel antibodies indicating probable Lambert-Eaton myasthenic syndrome. 75% of the children, compared with only 28% of the adults, had ocular MG. Thymoma was evident by MRI in 1.5% of children and in 20% of adults. Despite most patients having received prednisone, very few had obtained full clinical remission. CONCLUSION: This study emphasises the frequency of early childhood onset with ocular symptoms and shows that many of these patients have AChR antibodies. By contrast, patients presenting in later age seem to be very uncommon in comparison with recent studies in Caucasian populations.

Adolescent↗

Comparative study on central metabolic fluxes of Bacillus megaterium strains in continuous culture using 13C labelled substrates.

Fluxes of central carbon metabolism [glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid cycle (TCA cycle), biomass formation] were determined for several Bacillus megaterium strains (DSM319, WH320, WH323, MS941) in C- and N-limited chemostat cultures by (13)C labelling experiments. The labelling patterns of proteinogenic amino acids were analysed by GC/MS and therefrom flux ratios at important nodes within the metabolic network could be calculated. On the basis of a stoichiometric metabolic model flux distributions were estimated for the different B. megaterium strains used at various cultivation conditions. Generally all strains exhibited similar metabolic flux distributions, however, several significant changes were found in (1) the glucose flux entering the PPP via the oxidative branch, (2) the reversibilities within the PPP, (3) the relative fluxes of pyruvate and acetyl-CoA fed to the TCA cycle, (4) the fluxes around the pyruvate node involving a futile cycle.

Bacillus megaterium↗

Quantitative determination of ginsenoside Rh2 in rat biosamples by liquid chromatography electrospray ionization mass spectrometry.

Ginsenoside Rh2 is a "hot" natural compound with great potential as a new anti-cancer drug based on abundant pharmacological experiments. However, no systemic pharmacokinetic study of Rh2 was reported because current analysis methods could not fully meet the requirements. Thus, we developed a simple LC/MS method with highly improved sensitivities for the determination of Rh2 in rat plasma, bile, urine, feces and most tissues. The tissues and feces were firstly homogenized mechanically using buffer and methanol as the media, respectively. Plasma, bile, urine and tissue homogenates were extracted with diethyl ether for sample preparation. Feces homogenates were directly deproteinized with acetonitrile. The subsequent analysis procedures were performed on a Shimadzu LCMS2010A system (electrospray ionization single quadrupole mass analyzer), with an ODS column (150 mm x 2.0-mm i.d., 5 microm) plus a C18 guard column for separation and ammonium chloride (500 micromol) as mobile phase additive. The proportions of mobile phase were changed timely according to gradient programs. Chlorinated adducts of molecular ions [M + Cl]- of Rh2 at m/z 657.35 and internal standard digitoxin at m/z 799.55 were monitored in selective ion monitoring mode of negative ions. The method was validated to be accurate, precise and rugged with good linearity in all matrices, according to the FDA guidelines. The lower limits of quantitation in rat plasma, urine and feces were 0.2, 0.2 and 20 ng/mL respectively. Stability studies were also performed, indicating that there were no stability-related problems in the analytical procedure of Rh2. The proposed method was successfully applied to the preclinical pharmacokinetic research of Rh2 in rats, including plasma kinetics, tissue distribution and excretion studies.

Animals↗

Glycogen metabolism in tissues from a mouse model of Lafora disease.

Laforin, encoded by the EPM2A gene, by sequence is a member of the dual specificity protein phosphatase family. Mutations in the EPM2A gene account for around half of the cases of Lafora disease, an autosomal recessive neurodegenerative disorder, characterized by progressive myoclonus epilepsy. The hallmark of the disease is the presence of Lafora bodies, which contain polyglucosan, a poorly branched form of glycogen, in neurons, muscle and other tissues. Glycogen metabolizing enzymes were analyzed in a transgenic mouse over-expressing a dominant negative form of laforin that accumulates Lafora bodies in several tissues. Skeletal muscle glycogen was increased 2-fold as was the total glycogen synthase protein. However, the -/+glucose-6-P activity of glycogen synthase was decreased from 0.29 to 0.16. Branching enzyme activity was increased by 30%. Glycogen phosphorylase activity was unchanged. In whole brain, no differences in glycogen synthase or branching enzyme activities were found. Although there were significant differences in enzyme activities in muscle, the results do not support the hypothesis that Lafora body formation is caused by a major change in the balance between glycogen elongation and branching activities.

Animals↗

Experimental therapy of prostate cancer with an immunomodulatory oligonucleotide: effects on tumor growth, apoptosis, proliferation, and potentiation of chemotherapy.

BACKGROUND: The present study was designed to demonstrate the therapeutic efficacy of a novel immunomodulatory oligonucleotide (IMO) for prostate cancer. METHODS: We evaluated the effects of the IMO in xenograft (PC-3) and syngeneic (TRAMP C1) models of prostate cancer, and in prostate cancer cells. The IMO was also evaluated in combination with chemotherapy, and the in vitro expression of TLR9 was examined. RESULTS: The IMO had significant anti-tumor activity in both prostate cancer models and almost complete tumor regression was observed when the IMO was combined with taxotere or gemcitabine. TLR9 mRNA and protein were both expressed in prostate cancer cells. The IMO also induced apoptosis and decreased proliferation and survival of PC-3 cells in vitro in the presence of Lipofectin. CONCLUSIONS: The IMO inhibits prostate cancer growth in vivo and in vitro, and potentiates the effects of conventional chemotherapeutic agents. This is the first report of TLR9 expression in prostate cancer cells.

Adenocarcinoma↗

Modification of poly(dimethylsiloxane) microfluidic channels with silica nanoparticles based on layer-by-layer assembly technique.

A hydrophilic poly(dimethylsiloxane) (PDMS) microchip with stable electroosmotic flow (EOF) was prepared by a simple and reproducible coating procedure with silica nanoparticles. The microchannel wall of PDMS chip was coated with a layer of poly(diallyldimethylammonium chloride) (PDDA) and then collected silica nanoparticles. The assembly was followed by contact angle, charge-coupled device (CCD) imaging, electroosmotic flow (EOF) measurements and electrophoretic separation experiments. Contact angle measurements revealed the coated surface was hydrophilic; the water contact angle for coated chips was 64 degrees compared with a water contact angle for native PDMS chips of 113 degrees . CCD images indicated a substantially more hydrophilic microchannel than native PDMS. We carried out a comparison and concluded that the EOF values on the coated PDMS chip were close to those values on the glass chip above pH 7.0. The coated channel had an excellent stability and reproducibility, RSD of EOF values (n=6) on native and coated PDMS microchip was 1.58 and 0.57%, respectively. Separation of dopamine and epinephrine was performed on the coated chip generated 1.40 x 10(5), 1.39 x 10(5) theoretical plates/m compared with the native PDMS chip of 0.79 x 10(5), 0.88 x 10(5), high resolution of 1.7 was achieved with a channel of 3.60 cm length.

Dimethylpolysiloxanes↗

Characterization of E. coli O24 and O56 O antigen gene clusters reveals a complex evolutionary history of the O24 gene cluster.

O antigen is part of the lipopolysaccharide present in the outer membrane of Gram-negative bacteria. It has many different forms, which are almost entirely due to genetic variations of O antigen gene clusters. In this study, the O antigen gene clusters of E. coli O24 and O56 were sequenced, and all genes were assigned functions on the basis of homology. Comparison of O antigen gene clusters indicated that E. coli O24 O antigen gene cluster has possibly arisen from the E. coli O56 gene cluster, through inactivation of two glycosyltransferase genes and acquisition of two new genes from E. coli O157 and O152, respectively. The insertion sequence elements seemed to play important roles for the assembly of the O24 O antigen gene cluster. This is the first time that the evolutionary history of a multi-origin O antigen gene cluster is clearly demonstrated. Genes specific to E. coli O24 and O56 were also identified, which may be used for development of DNA-based serotyping schemes.

Base Sequence↗

Capillary zone electrophoresis method for fingerprint of allantoic fluid in normal and infected SPF embryonated chicken eggs.

A capillary zone electrophoresis method was developed for fingerprint determination of allantoic fluid in specific pathogen free (SPF) embryonated chicken eggs. The effects of some crucial parameters, such as buffer type, pH, wavelength and running voltage on the separation were studied systematically. The components of the allantoic fluid were well separated using a fused-silica uncoated capillary with an effective length of 50 cm and an internal diameter of 50 microm. One hundred millimolars sodium tetraborate buffer containing 20 mM sodium dihydrogen phosphate with a final pH 9.8 was used as a running buffer. Comparative fingerprints of allantoic fluid in normal and infected with infectious bronchitis virus (IBV) SPF embryonated chicken eggs were also evaluated. The results showed that there were significant differences between composition of normal allantoic fluid and allantoic fluid infected with IBV, which led to different migration behavior. This method was shown to be stable and reproducible with a relative standard deviation of less than 5% for both migration time and peak current.

Allantois↗

A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus.

The spike (S) protein of SARS coronavirus (SARS-CoV) is responsible for viral binding with ACE2 molecules. Its receptor-binding motif (S-RBM) is located between residues 424 and 494, which folds into 2 anti-parallel beta-sheets, beta5 and beta6. We have previously demonstrated that fragment 450-650 of the S protein (S450-650) is predominantly recognized by convalescent sera of SARS patients. The N-terminal 60 residues (450-510) of the S450-650 fragment covers the entire beta6 strand of S-RBM. In the present study, we demonstrate that patient sera predominantly recognized 2 linear epitopes outside the beta6 fragment, while the mouse antisera, induced by immunization of BALB/c mice with recombinant S450-650, mainly recognized the beta6 strand-containing region. Unlike patient sera, however, the mouse antisera were unable to inhibit the infectivity of S protein-expressing (SARS-CoV-S) pseudovirus. Fusion protein between green fluorescence protein (GFP) and S450-650 (S450-650-GFP) was able to stain Vero E6 cells and deletion of the beta6 fragment rendered the fusion product (S511-650-GFP) unable to do so. Similarly, recombinant S450-650, but not S511-650, was able to block the infection of Vero E6 cells by the SARS-CoV-S pseudovirus. Co-precipitation experiments confirmed that S450-650 was able to specifically bind with ACE2 molecules in lysate of Vero E6 cells. However, the ability of S450-510, either alone or in fusion with GFP, to bind with ACE2 was significantly poorer compared with S450-650. Our data suggest a possibility that, although the beta6 strand alone is able to bind with ACE2 with relatively high affinity, residues outside the S-RBM could also assist the receptor binding of SARS-CoV-S protein.

Angiotensin-Converting Enzyme 2↗

Oroxylin A induced apoptosis of human hepatocellular carcinoma cell line HepG2 was involved in its antitumor activity.

We previously reported that wogonin, a flavonoid compound, was a potent apoptosis inducer of human hepatoma SMMC-7721 cells and murine sarcoma S180 cells. In the present study, the effect of oroxylin A, one wogonin structurally related flavonoid isolated from Scutellariae radix, on human hepatocellular carcinoma cell line HepG2 was examined and molecular mechanisms were also investigated. Oroxylin A inhibited HepG2 cell proliferation in a concentration- and time-dependent manner measured by MTT-assay. Treatment with an apoptosis-inducing concentration of oroxylin A caused typical morphological changes and apoptotic blebbing in HepG2 cells. DNA fragmentation assay was used to examine later apoptosis induced by oroxylin A. FACScan analysis revealed a dramatic increase in the number of apoptotic and G(2)/M phase arrest cells after oroxylin A treatment. The pro-apoptotic activity of oroxylin A was attributed to its ability to modulate the concerted expression of Bcl-2, Bax, and pro-caspase-3 proteins. The expression of Bcl-2 protein and pro-caspase-3 protein was dramatically decreased after treatment with oroxylin A. These results demonstrated that oroxylin A could effectively induce programmed cell death and suggested that it could be a promising antitumor drug.

Antineoplastic Agents↗

Multiple perspectives on diagnosis delay for tuberculosis from key stakeholders in poor rural China: case study in four provinces.

This study aims to understand the contextual barriers to accessing timely TB diagnosis after first seeking care, especially among the poor and vulnerable in rural China. Both quantitative and qualitative methods were used to elicit the experiences and perspectives of TB patients and suspected TB patients, community residents, health providers and policy makers in poor, rural areas of four provinces. Between 30 and 60% of patients across the four provinces experienced a delay in receiving a diagnosis after first seeking care. Most patients had to visit health facilities more than once before diagnosis, with 17-30% patients making more than 6 visits. These delays and multiple visits mainly occurred because of the limited capacity of health providers to recognize TB, and financial disincentives to refer patients to TB dispensaries, due to the pressures of the cost recovery system. Poverty and socio-economic disadvantage amongst patients also influenced their capability to seek further care to obtain a reliable diagnosis. Qualitative data showed that women and the elderly patients were likely to experience more 'system' delay, and these findings were to some extent supported by the survey. The study concludes that 'system' delay is a serious problem, which is influenced by the financing mechanisms for both TB control and general health services as well as poverty and disadvantage amongst patients. This requires a comprehensive strategy to shorten 'system' delay in order to enable successful DOTS expansion, including developing appropriate financing mechanisms to improve general provider capacity and encourage referral, as well as measures to improve financial and social access to services for potential TB patients.

Adolescent↗

AT1 receptors in the nucleus tractus solitarii mediate the interaction between the baroreflex and the cardiac sympathetic afferent reflex in anesthetized rats.

The cardiac "sympathetic afferent" reflex (CSAR) has been reported to increase sympathetic outflow and depress baroreflex function via a central angiotensin II (ANG II) mechanism. In the present study, we examined the role of ANG II type 1 (AT(1)) receptors in the nucleus tractus solitarii (NTS) in mediating the interaction between the CSAR and the baroreflex in anesthetized rats. We examined the effects of bilateral microinjection of AT(1) receptor antagonist losartan (100 pmol) into the NTS on baroreflex control of renal sympathetic nerve activity (RSNA) before and after CSAR activation by epicardial application of capsaicin (0.4 microg). Using single-unit extracellular recording, we further examined the effects of CSAR activation on the barosensitivity of barosensitive NTS neurons and the effects of intravenous losartan (2 mg/kg) on CSAR-induced changes in activity of NTS barosensitive neurons. Bilateral NTS microinjection of losartan significantly attenuated the increases in arterial pressure, heart rate, and RSNA evoked by capsaicin but also markedly (P < 0.01) reversed the CSAR-induced blunted baroreflex control of RSNA (Gain(max) from 1.65 +/- 0.10 to 2.22 +/- 0.11%/mmHg). In 17 of 24 (70.8%) NTS barosensitive neurons, CSAR activation significantly (P < 0.01) inhibited the baseline neuronal activity and attenuated the neuronal barosensitivity. In 11 NTS barosensitive neurons, intravenous losartan effectively (P < 0.01) normalized the decreased neuronal barosensitivity induced by CSAR activation. In conclusion, blockade of NTS AT(1) receptors improved the blunted baroreflex during CSAR activation, suggesting that the NTS plays an important role in processing the interaction between the baroreflex and the CSAR via an AT(1) receptor-dependent mechanism.

Analgesics, Non-Narcotic↗

Comparison of the acute toxicity for gamma-cyhalothrin and lambda-cyhalothrin to zebra fish and shrimp.

Gamma-cyhalothrin 15CS (GCH) contains only the active stereoisomer of the two isomers found in lambda-cyhalothrin 25EW (LCH). GCH (0.5 x rate) provides equivalent overall insect control as LCH (1 x rate). Both formulations showed high acute toxicity to zebra fish (Brachydanio rerio H.B.) and shrimp (Macrobrachium nippoensis de Haan). The 96-h LC(50(zebra fish,GCH)) is 1.93 microg a.i/L and LC(50(zebra fish,LCH)) is 1.94 microg a.i/L. LC(50(shrimp,GCH)) is 0.28 microg a.i./L and LC(50(shrimp,LCH)) 0.04 microg a.i./L. This indicates that the toxicity to shrimp is likely stereochemistry-dependent. The fates of GCH and LCH are similar in laboratory simulated rice paddy water and their concentrations decrease rapidly, with no GCH or LCH detected after 3 or 4 days. Both are toxic to shrimp in a simulated paddy irrigation reservoir even though treated return water is diluted 5 times. No shrimp fatality is shown in the GCH-treated paddy water after a 4-day holding period, and longer than 5 days is necessary to reach a zero fatality rate for LCH. This is compatible with the 7-day water holding period considered reasonable in agricultural practice.

Animals↗

Unique aggregation of anthrax (Bacillus anthracis) spores by sugar-coated single-walled carbon nanotubes.

There has been significant interest in the binding of anthrax spores by molecular species, but with only limited success. Proteins and more recently peptides were used. However, despite the known presence of carbohydrates on the spore surface, carbohydrate-carbohydrate interactions have hardly been explored likely because of the lack of required specific platform for synthetic carbohydrates. We report the successful use of single-walled carbon nanotubes as a truly unique scaffold for displaying multivalent monosaccharide ligands that bind effectively to anthrax spores with divalent cation mediation to cause significant spore aggregation. The work should have far-reaching implications in development of countermeasure technologies.

Animals↗