Biomedical subjects

Feng Li

Publications and source records attributed to Feng Li.

At least 163 records · Page 9Linked to original sources

PubMed2004

Identification and characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1.

Nasopharyngeal carcinoma (NPC) is one of the most commons cancers in Southeast Asia and Southern China. Several NPC-associated genes have been so far described and here we describe the identification and the characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1. NAP-1 was identified with the human genome draft searching method combined with nested PCR mapping of the chromosome 4q13 region. NAP-1 encodes an 85 amino acid alkaline peptide with a calculated isoelectric point of 9.3, three phosphorilation sites and a proline-rich region. Northern blot analysis revealed that NAP-1 is expressed as a 0.6 kb transcript in normal lymph nodes and trachea. In addition, reverse transcription (RT)-PCR showed that NAP-1 is expressed not only in NPC but in normal nasopharynx (NP) and various other tumors and tissues of the head and neck including: tonsils, lymph nodes, carcinoma of the tonsil, T cell lymphomas, squamous cell carcinoma of the hard palate, papilloma of the nasopharynx, nasopharyngitis, lymphoma of the tongue root and follicular dendritic cells (FDC). In addition, NAP-1 is not expressed in normal tissues or tumors from other anatomical regions and was not expressed by NPC cell lines. Surprisingly, differential RT-PCR demonstrated decreased expression of NAP-1 in NPC compared with paired NP biopsies in 42.5 % of cases (17 out of 40). In addition, in situ hybridization and immunohistochemistry demonstrated that NAP-1 is expressed by S100+ CD35+ FDCs of the germinal center and not in other normal immune cells infiltrating NP or NPC. Therefore, it is likely that NAP-1 is secreted by FDC in the NP and may play an immune modulatory role in NPC.

Journal Article↗

PubMed2004

[Effects of the escharectomy during burn shock stage on expression of glucose translator-4 mRNA in skeletal muscle and adipose tissue].

OBJECTIVE: To investigate the effect of the escharectomy during burn shock stage on expression of glucose translator-4 (GLUT4) mRNA in skeletal muscle and adipose tissue. METHODS: 30% TBSA scalded rats were employed. Escharectomy were conducted at 8 h, 24 h, 168 h after burns respectively. Insulin, glucagon, cortisol and glucose levels in serum were analyzed. RT-PCR were employed to analyze GLUT4 mRNA expression in skeletal muscle and adipose tissue. RESULTS: Glucagon, cortisol and glucose levels in serum were declined in groups which escharectomy were conducted during burn shock stage. GLUT4 mRNA expression in both skeletal muscle and adipose tissue were downregulated after burns and escharectomy conducted during burn shock stage made it restored to near normal. CONCLUSION: GLUT4 mRNA expression will declined after major burns in skeletal muscle and adipose tissue. Escharectomy during shock stage could make it upregulated, which will be helpful to improve glucose metabolism and hypermetabolism after major burns.

Adipose Tissue↗

PubMed2004

P23H and S334ter opsin mutations: Increasing photoreceptor outer segment n-3 fatty acid content does not affect the course of retinal degeneration.

PURPOSE: The n-3 polyunsaturated fatty acids (PUFA) facilitate retinal development and function. Rats carrying transgenes with P23H and S334ter rhodopsin mutations lose their photoreceptors and have lower levels of 22:6n-3 in rod photoreceptor outer segments (ROS) than wild type (WT) animals. We tested the hypothesis that the rate of retinal degeneration in these mutant animals could be sensitive to the n-3 fatty acid content of retina. METHODS: Beginning embryonic day 15, WT and heterozygous transgenic rats with P23H and S344ter rhodopsin mutations were fed semi-synthetic diets enriched in n-6 (safflower oil, SO) or n-3 (flaxseed oil, FO) PUFA. At 35 and 55 days of age, electroretinographic (ERG) response, outer nuclear layer (ONL) thickness, and fatty acid composition of plasma and ROS were determined. Student's t-tests and multivariate analysis of variance with post hoc tests determined statistical differences. RESULTS: Rats fed FO or SO diets had different n-6/n-3 PUFA ratios in plasma (1.3 and 62) and ROS (0.2 and 1.1, respectively). Although there were profound effects of the diets on the plasma fatty acid composition, there were only minor differences between WT and transgenic animals within each dietary regime. The ROS of FO fed rats had 70% more 22:6n-3 than those fed SO, and the WT had higher concentrations of 22:6n-3 than the transgenic animals (WT>P23H>S334ter). In contrast, there was no difference in 22:6n-3 levels in ROS of WT and transgenic rats fed the SO diet. At P55, both transgenic lines had diminished ERGs and ONL thickness relative to the WT. There was no detectable effect of ROS fatty acid enrichment on the rate of retinal degeneration in the transgenic animals. However, the FO-diet provided a modest protection of function (b-wave) in S334ter animals. CONCLUSIONS: Feeding n-3 fatty acids to rats with mutant rhodopsin transgenes significantly increased the levels of 22:6n-3 in ROS membranes, but had no effect on the rate of retinal degeneration. Therefore, the degeneration is not the result of low (or high) 22:6n-3 in ROS and supplementation with 18:3n-3 will not rescue dying photoreceptor cells in these animal models of inherited retinal degenerations.

Animals↗

PubMed2004

Effect of nonionic fluorosurfactant on the electrogenerated chemiluminescence of the tris(2,2'-bipyridine)ruthenium(II)/tri-n-propylamine system: lower oxidation potential and higher emission intensity.

Fluorosurfactants are commercially available, and their applications in electrochemical systems have been the interest of many studies. Here, we describe a novel effect of a nonionic fluorosurfactant (Zonyl FSN) on the electrogenerated chemiluminescence (ECL) of the tris(2,2'-bipyridine)ruthenium(II)/tri-n-propylamine (TPrA) system at gold and platinum electrodes. Compared with its hydrocarbon analogue (Triton X-100), the adsorbed fluorosurfactant species not only rendered the electrode surfaces more hydrophobic but also significantly retarded the growth of the electrode oxide layers. As a result, more facile direct oxidation of TPrA was achieved, which led to the appearance of a low oxidation potential ECL signal (below 1.0 V vs SCE). At the gold electrode, the ECL peak appeared at 0.82 V, approximately 400 mV more negative than usual; while its intensity was approximately 50 times higher. The generation of the intense ECL signal at low oxidation potential may lead to the development of more efficient ECL analysis.

Journal Article↗

PubMed2004

Flow injection analysis of gallic acid with inhibited electrochemiluminescence detection.

A flow injection (FI)-electrochemiluminescent (ECL) method has been developed for the determination of gallic acid, based on an inhibition effect on the Ru(bpy)(3)(2+)/tri- n-propylamine (TPrA) ECL system in pH 8.0 phosphate buffer solution. The method is simple and convenient with a determination limit of 9.0x10(-9) mol/L and a dynamic concentration range of 2x10(-8)-2x10(-5) mol/L. The relative standard deviation (RSD) was 1.0% for 1.0x10(-6) mol/L gallic acid ( n=11). It was successfully applied to the determination of gallic acid in Chinese proprietary medicine-Jianming Yanhou Pian. The inhibition mechanism proposed for the quenching effect of the gallic acid on the Ru(bpy)(3)(2+)/TPrA ECL system was the interaction of electrogenerated Ru(bpy)(3)(2+*) and o-benzoquinone derivative at the electrode surface. The ECL emission spectra and UV-visible absorption spectra were applied to confirm the mechanism.

Buffers↗

PubMed2004

Interferon antagonist proteins of influenza and vaccinia viruses are suppressors of RNA silencing.

Homology-dependent RNA silencing occurs in many eukaryotic cells. We reported recently that nodaviral infection triggers an RNA silencing-based antiviral response (RSAR) in Drosophila, which is capable of a rapid virus clearance in the absence of expression of a virus-encoded suppressor. Here, we present further evidence to show that the Drosophila RSAR is mediated by the RNA interference (RNAi) pathway, as the viral suppressor of RSAR inhibits experimental RNAi initiated by exogenous double-stranded RNA and RSAR requires the RNAi machinery. We demonstrate that RNAi also functions as a natural antiviral immunity in mosquito cells. We further show that vaccinia virus and human influenza A, B, and C viruses each encode an essential protein that suppresses RSAR in Drosophila. The vaccinia and influenza viral suppressors, E3L and NS1, are distinct double-stranded RNA-binding proteins and essential for pathogenesis by inhibiting the mammalian IFN-regulated innate antiviral response. We found that the double-stranded RNA-binding domain of NS1, implicated in innate immunity suppression, is both essential and sufficient for RSAR suppression. These findings provide evidence that mammalian virus proteins can inhibit RNA silencing, implicating this mechanism as a nucleic acid-based antiviral immunity in mammalian cells.

Animals↗

PubMed2004

p41-Arc subunit of human Arp2/3 complex is a p21-activated kinase-1-interacting substrate.

The formation of new branched actin filament networks at the cell cortex of migrating cells is choreographed by the actin-related protein (Arp) 2/3 complex. Despite the fundamental role of the Arp2/3 complex in actin nucleation and branching, upstream signals that control the functions of p41-Arc, a putative regulatory component of the mammalian Arp2/3 complex, remain unidentified. Here we show that p41-Arc interacts with p21-activated kinase 1 (Pak1) both in vitro and in vivo. Pak1 phosphorylation of p41-Arc regulates its localization with the Arp2/3 complex in the cortical nucleation regions of cells. Pak1 phosphorylates p41-Arc on threonine 21 in the first WD repeat, and its mutation has functional implications in vivo. Threonine 21 phosphorylation by Pak1 is required for both constitutive and growth-factor-induced cell motility. Pak1 regulation of p41-Arc activation status represents a novel mechanism by which signalling pathways may influence the functions of the Arp2/3 complex, leading to motility in mammalian cells.

Actin Cytoskeleton↗

PubMed2004

A flexible peptide linker enhances the immunoreactivity of two copies HBsAg preS1 (21-47) fusion protein.

PreS1 (21-47) region of HBV large surface protein is hepatocyte receptor binding site and the anti-preS1 (21-47) antibody possesses the virus-neutralizing activity and protective effect. It is important to obtain the peptide with higher immunoreactivity on a large scale for detecting the anti-preS1 (21-47) antibody in the sera from HBV infected patients and future vaccine recipients. The expression vector pGEX SLS, which expressed two copies of the preS1 (21-47) peptide connected by a flexible linker (Gly4Ser3) fused to glutathione S-transferase (GST), was constructed. The fusion protein, named GST-SLS, was highly expressed in E. coli and purified by affinity chromatography. Ninety milligrams purified protein can be obtained from 1l of culture. The data in ELISA analysis showed that the immunoreactivity of GST-SLS was enhanced significantly in comparison with GST-S II, a GST fusion protein with two copies preS1 (21-47) linked directly; GST-S I, another GST fusion protein with one copy preS1 (21-47) and preS1 (21-47) synthesized peptide. In addition, GST-SLS has been tried to use in detecting anti-preS1 (21-47) antibody in the sera from HBV infected patients and a satisfied result was gained. Therefore, GST-SLS may have potential to be developed into a new kit for diagnosis and prognosis of hepatitis B (HB) patients.

Enzyme-Linked Immunosorbent Assay↗

PubMed2004

Involvement of insulin/phosphoinositide 3-kinase/Akt signal pathway in 17 beta-estradiol-mediated neuroprotection.

In the present study, we tested the hypothesis that 17beta-estradiol (betaE2) is a neuroprotectant in the retina, using two experimental approaches: 1) hydrogen peroxide (H(2)O(2))-induced retinal neuron degeneration in vitro, and 2) light-induced photoreceptor degeneration in vivo. We demonstrated that both betaE2 and 17alpha-estradiol (alphaE2) significantly protected against H(2)O(2)-induced retinal neuron degeneration; however, progesterone had no effect. betaE2 transiently increased the phosphoinositide 3-kinase (PI3K) activity, when phosphoinositide 4,5-bisphosphate and [(32)gammaATP] were used as substrate. Phospho-Akt levels were also transiently increased by betaE2 treatment. Addition of the estrogen receptor antagonist tamoxifen did not reverse the protective effect of betaE2, whereas the PI3K inhibitor LY294002 inhibited the protective effect of betaE2, suggesting that betaE2 mediates its effect through some PI3K-dependent pathway, independent of the estrogen receptor. Pull-down experiments with glutathione S-transferase fused to the N-Src homology 2 domain of p85, the regulatory subunit of PI3K, indicated that betaE2 and alphaE2, but not progesterone, identified phosphorylated insulin receptor beta-subunit (IRbeta) as a binding partner. Pretreatment with insulin receptor inhibitor, HNMPA, inhibited IRbeta activation of PI3K. Systemic administration of betaE2 significantly protected the structure and function of rat retinas against light-induced photoreceptor cell degeneration and inhibited photoreceptor apoptosis. In addition, systemic administration of betaE2 activated retinal IRbeta, but not the insulin-like growth factor receptor-1, and produced a transient increase in PI3K activity and phosphorylation of Akt in rat retinas. The results show that estrogen has retinal neuroprotective properties in vivo and in vitro and suggest that the insulin receptor/PI3K/Akt signaling pathway is involved in estrogen-mediated retinal neuroprotection.

Animals↗

PubMed2004

Computerized scheme for automated detection of lung nodules in low-dose computed tomography images for lung cancer screening.

RATIONALE AND OBJECTIVES: A computerized scheme for automated detection of lung nodules in low-dose computed tomography images for lung cancer screening was developed. MATERIALS AND METHODS: Our scheme is based on a difference-image technique for enhancing the lung nodules and suppressing the majority of background normal structures. The difference image for each computed tomography image was obtained by subtracting the nodule-suppressed image processed with a ring average filter from the nodule-enhanced image with a matched filter. The initial nodule candidates were identified by applying a multiple-gray level thresholding technique to the difference image, where most nodules were well enhanced. A number of false-positives were removed first in entire lung regions and second in divided lung regions by use of the two rule-based schemes on the localized image features related to morphology and gray levels. Some of the remaining false-positives were eliminated by use of a multiple massive training artificial neural network trained for reduction of various types of false-positives. This computerized scheme was applied to a confirmed cancer database of 106 low-dose computed tomography scans with 109 cancer lesions for 73 patients obtained from a lung cancer screening program in Nagano, Japan. RESULTS: This computed-aided diagnosis scheme provided a sensitivity of 83% (91/109) for all cancers with 5.8 false-positives per scan, which included 84% (32/38) for missed cancers with 5.9 false-positives per scan. CONCLUSION: This computerized scheme may be useful for assisting radiologists in detecting lung cancers on low-dose computed tomography images for lung cancer screening.

False Positive Reactions↗

PubMed2004

Effect of temporal subtraction images on radiologists' detection of lung cancer on CT: results of the observer performance study with use of film computed tomography images.

RATIONALE AND OBJECTIVES: To evaluate the effect of temporal subtraction images on the radiologists' detection of early primary lung cancer in computed tomography (CT) scans. MATERIALS AND METHODS: Fourteen cases with primary lung cancer and 16 normal cases were used for this study from a database of low-dose CT images, which were obtained from a lung cancer screening program in Nagano, Japan. Images were obtained with a single-detector helical CT scanner using 10 mm collimation and 2:1 pitch. Each case had both previous and current CT scans. Temporal subtraction images were obtained by subtracting the warped previous images from the current images. Seven radiologists, including four attendings and three residents, provided their confidence levels for the presence or absence of lung cancers with use of film CT images without and with temporal subtraction images. Receiver operating characteristic analysis was used to compare their performance without and with temporal subtraction images. RESULTS: The mean Az values (area under the receiver operating characteristic curve) of seven observers without and with temporal subtraction images were 0.868 and 0.930, respectively. Diagnostic accuracy was significantly improved by using temporal subtraction images (P = .007). Temporal subtraction images were especially useful when a nodule was present near the pulmonary hilum, where radiologists tended to overlook it. CONCLUSION: The temporal subtraction technique can significantly improve the sensitivity and specificity for detection of lung cancer on CT scans.

Adult↗

PubMed2004

Influence of MRI abnormality in skull base bone on prognosis of nasopharyngeal carcinoma.

PURPOSE: To evaluate the influence of skull base bone (SBB) abnormality showed by MRI on prognosis of nasopharyngeal carcinoma (NPC). PATIENTS AND METHODS: From March 1993 to December 1998, 122 NPC patients received prime radiotherapy treatment. All of them were proved pathologically and checked by magnetic resonance imaging (MRI). Every patient received radiation through conjoint facio-cervical field and conventional dose-fractionation schedules. The total dose to the primary tumor was 60-75 Gy (median, 70 Gy). The Kaplan-Meier method, the Log-rank test and the Cox regression model were used to evaluate the significance of prognostic factors on NPC patient survival. RESULTS: The overall median survival period was 50 (6-92) months, and the 1, 3 and 5 year-survival rates were, respectively, 99.2%, 87.9%, and 73.3%. The 1, 3, and 5 year-survival rates of abnormality and normality of the SBB on MRI were 98.9%, 87.2%, 71.9%, and 100.0%, 89.8%, 77.0%, respectively (P = 0.4233). Gender, age, head pain, SBB abnormality, cranial nerve palsy, cervical lymphadenopathy and primary tumor extent were analyzed with the Cox regression model and SBB abnormality on MRI did not prove to have statistical significance (P = 0.6934). According to the analysis of regrouping, patients with SBB abnormalities > or =2 sites have a worse prognosis (P = 0.0427). Then, the above seven factors are analyzed by Cox regression model and the result had statistical significance (P = 0.0385). CONCLUSION: The SBB abnormality on MRI is of no obvious influence on prognosis of NPC. However, when SBB abnormality sites were > or =2, there is obvious statistical significance on the prognosis.

Adolescent↗

PubMed2004

Artificial neural networks (ANNs) for differential diagnosis of interstitial lung disease: results of a simulation test with actual clinical cases.

RATIONALE AND OBJECTIVES: To evaluate the performance of an artificial neural network (ANN) scheme with use of consecutive clinical cases and its effect on radiologists with an observer test. MATERIALS AND METHODS: Artificial neural networks were designed to distinguish among 11 interstitial lung diseases on the basis of 26 inputs (16 radiologic findings, 10 clinical parameters). Chest radiographs of 96 consecutive cases with interstitial lung disease were used. Five radiologists independently rated their radiologic findings on the 96 chest radiographs. Based on their ratings of radiologic findings and clinical parameters obtained from the hospital information system, the output values indicating the likelihood of each of the 11 interstitial lung diseases were determined. Subsequently, 30 cases were selected from these 96 cases for an observer test. Five radiologists marked their confidence levels for diagnosis of 11 possible diseases in each case without and with ANN output. The performance of ANNs and radiologists was evaluated by receiver operating characteristic analysis based on their outputs and on confidence levels, respectively. RESULTS; The average Az value (area under the receiver operating characteristic curve) indicating ANN performance for the 96 consecutive cases was 0.85 +/- 0.03. The average Az values indicating radiologists' performance without and with ANN outputs were 0.81 +/- 0.11 and 0.87 +/- 0.06, respectively. The diagnostic accuracy was improved significantly when radiologists read chest radiographs with ANN outputs (P < .05). CONCLUSION: Artificial neural networks for differential diagnosis of interstitial lung disease may be useful in clinical situations, and radiologists may be able to utilize the ANN output to their advantage in the differential diagnosis of interstitial lung disease on chest radiographs.

Adult↗

PubMed2004

Shear stress-induced binding of large and unusually large von Willebrand factor to human platelet glycoprotein Ibalpha.

Platelet membrane glycoprotein (GP) Ib(alpha), a component of the GP Ib-IX-V complex, is a receptor for von Willebrand factor (VWF). A small quantity of large VWF multimers binds to platelets under high shear stress, and induces aggregation. We studied the shear-induced attachment of large and unusually large VWF multimers to the GPIb(alpha) extracellular domain (glycocalicin), human platelets, and GPIb(alpha) gxpressing Chinese hamster ovary (CHO) cells. Compared to binding in the presence of botrocetin and ristocetin, shear stress only induced low-level NVWF (normal plasma VWF multimers) binding. This shear stress induced interaction is also dependent on VWF multimeric size. Elevated binding levels of endothelial cell VWF (enriched in unusually large VWF multimers) to glycocalicin-coated beads were observed under low shear conditions, which did not result in the attachment of normal plasma VWF.

Animals↗

PubMed2004

Suppression of bcl-2 gene by RNA interference increases chemosensitivity to cisplatin in nasopharyngeal carcinoma cell line CNE1.

To explore the effect of suppressing BCL-2 expression using RNA interference (RNAi) technique in nasopharyngeal carcinoma cell line CNE1. CNE1 cell lines stably expressing shRNAs targeted bcl-2 and GL3 gene were established and gene expression inhibition was assessed by Western blotting analysis. The effect of suppressing bcl-2 by RNAi on cell growth was studied, the apoptosis induction and the sensitization of CNE1 cells to cisplatin were quantified by MTT assays and flow cytometry. The results showed that: stable transfection of CNE1 cells with vectors expressing shRNAs against bcl-2 decreased the expression of BCL-2 protein; suppression of BCL-2 expression did not affect cell proliferation but could increase the chemosensitivity to cisplatin in CNE1 cells. This will help physicians to make some clinical trials of gene therapy on nasopharyngeal carcinoma by RNAi.

Antineoplastic Agents↗

PubMed2004

Preparation and primary application of monoclonal antibodies against a novel ribosome-inactivating protein Moschatin from pumpkin seeds.

Plant ribosome-inactivating proteins (RIPs) have multiple biological functions, and have been widely used in the studies on biomedical and agronomic applications. Moschatin is a novel single-chain RIP recently purified from pumpkin seeds, and it has been successfully applied to construct the immunotoxin that can selectively kill the cultured human melanoma cells. Six stable strains of hybridomas (2H8, 4A8, 5B6, 6F8, 4H10 and 6C2) that can secrete high specific monoclonal antibodies against Moschatin have been successfully prepared using hybridoma technique. The isotypes of these monoclonal antibodies are IgG1, IgG1, IgG1, IgG1, IgG2a and IgGM. Their affinity constants were determined to be 1.42x10(8), 2.71x10(8), 8.72x10(7), 2.06x10(8), 1.36x10(8) and 1.51x10(8) M(-1) in a sequent order, measured by non-competitive ELISA. The monoclonal antibody 4A8 has been used to detect Moschatin in Western blot. An immunoaffinity gel, which consisted of a monoclonal antibody 4H10 and Sepharose 4B, was prepared and used to purify Moschatin from pumpkin seeds crude extract.

Antibodies, Monoclonal↗

PubMed2004

Expression of overlapping PreS1 fragment recombinant proteins for the determination of immunogenic domains in HBsAg PreS1 region.

In this study, eight preS1 fragments overlapped in preS1 (21-119) region of HBV adr subtype, i.e. preS1 (21-47), preS1 (34-59), preS1 (48-70), preS1 (60-85), preS1 (71-94), preS1 (86-109), preS1 (95-119) and preS1 (21-119), were cloned by PCR, and expressed as GST fusion proteins. These GSTpreS1 fusion proteins were highly expressed in soluble form in E. coli, and about 50 to 90 mg soluble fusion proteins were purified from 1 L culture. Using these fusion proteins, the immunogenic domains in preS1 (21-119) region were identified by Western blot analysis and competitive ELISA. The results showed that the immunogenic domains mainly existed in preS1 (21-59) in N-terminus and preS1 (95-109) in C-terminus, and more importantly, a major immunogenic domain preS1 (34-59), which has much stronger immunogenicity, was identified. It was also supported by the predictions of secondary structure and immunological property in the preS1 (21-119) region. The results here would be helpful for the design of new vaccines against HBV.

Animals↗

PubMed2004

Increased plaque burden in brains of APP mutant MnSOD heterozygous knockout mice.

A growing body of evidence suggests a relationship between oxidative stress and beta-amyloid (Abeta) peptide accumulation, a hallmark in the pathogenesis of Alzheimer's disease (AD). However, a direct causal relationship between oxidative stress and Abeta pathology has not been established in vivo. Therefore, we crossed mice with a knockout of one allele of manganese superoxide dismutase (MnSOD), a critical antioxidant enzyme, with Tg19959 mice, which overexpress a doubly mutated human beta-amyloid precursor protein (APP). Partial deficiency of MnSOD, which is well established to cause elevated oxidative stress, significantly increased brain Abeta levels and Abeta plaque burden in Tg19959 mice. These results indicate that oxidative stress can promote the pathogenesis of AD and further support the feasibility of antioxidant approaches for AD therapy.

Alzheimer Disease↗