Biomedical subjects

Lin Chen

Publications and source records attributed to Lin Chen.

At least 109 records · Page 6Linked to original sources

PubMed2005

Inhibitive effects of quercetin on rabbit tenon capsule fibroblasts proliferation.

PURPOSE: To study the inhibitive effects of quercetin (QU) on the fibroblasts proliferation of rabbit Tenon's capsule and its mechanism. METHODS: Cultured fibroblasts were exposed to different concentrations of QU solution and investigated by microculture tetrazolium (MTT) assay. The effect of QU was observed on cells cycle using the flow cytometer. RESULTS: QU can suppress the proliferation of rabbit Tenon's capsule fibroblasts in vitro and show a dose-time dependent tendency. Flow cytometer results showed 26.92% cell increase in G1 phase, 23.50% decrease in S phase and 3.42% decrease in G2 phase. CONCLUSIONS: QU can suppress the proliferation of rabbit Tenon's capsule fibroblasts in vitro and show a dose-time dependent tendency. QU may effect all phase of cell cycle and inhibit cell proliferation by inhibiting G1 phase transitting to S phase and G2 phase.

Animals↗

PubMed2004

Solution structure of prosurvival Mcl-1 and characterization of its binding by proapoptotic BH3-only ligands.

The B cell lymphoma-2 (Bcl-2) homologs myeloid cell leukemia-1 (Mcl-1) and A1 are prosurvival factors that selectively bind a subset of proapoptotic Bcl homology (BH) 3-only proteins. To investigate the molecular basis of the selectivity, we determined the solution structure of the C-terminal Bcl-2-like domain of Mcl-1. This domain shares features expected of a prosurvival Bcl-2 protein, having a helical fold centered on a core hydrophobic helix and a surface-exposed hydrophobic groove for binding its cognate partners. A number of residues in the binding groove differentiate Mcl-1 from its homologs, and in contrast to other Bcl-2 homologs, Mcl-1 has a binding groove in a conformation intermediate between the open structures characterized by peptide complexes and the closed state observed in unliganded structures. Mutagenesis of potential binding site residues was used to probe the contributions of groove residues to the binding properties of Mcl-1. Although mutations in Mcl-1 had little impact on binding, a single mutation in the BH3-only ligand Bad enabled it to bind both Mcl-1 and A1 while retaining its binding to Bcl-2, Bcl-xL, and Bcl-w. Elucidating the selective action of certain BH3-only ligands is required for delineating their mode of action and will aid the search for effective BH3-mimetic drugs.

Amino Acid Sequence↗

PubMed2004

Multi-phasic CT arterial portography and CT hepatic arteriography improving the accuracy of liver cancer detection.

AIM: To evaluate the value of multi-phasic CT arterial portography (CTAP) and CT hepatic arteriography (CTHA) in differential diagnosis of liver diseases, and to improve the specificity of CTAP and CTHA for liver cancer detection. METHODS: From January 1999 to December 2002, multi-phasic CTAP and CTHA were performed in 20 patients with suspected liver disease. CT scanning was begun 25 s, 60 s and 120 s for the early-, late- and delayed-phase CTAP examinations, and 6 sec, 40 s and 120 s for the early-, late- and delayed-phase CTHA examinations respectively, after a transcatheter arterial injection of non-ionic contrast material. If a lesion was diagnosed as a liver cancer, transcatheter hepatic arterial chemoembolization (TACE) treatment was performed, and the follow-up CT was performed three or four weeks later. RESULTS: All eighteen HCCs in 12 cases were shown as nodular enhancement on early-phasic CTHA. The density of the whole tumor decreased rapidly on late and delayed phases, and the edge of 12 tumors (12/18) remained relatively hyperdense compared with the surrounding liver tissue, and demonstrated as rim enhancement. All HCCs were shown as perfusion defect nodules on multi-phasic CTAP. Five tumors (5/18) were shown as rim enhancement on delayed-phasic CTAP. Rim enhancement was shown as 1 to 2-mm-wide irregular, uneven and discontinuous circumferential enhancement at late-, and delayed-phase of CTHA or CTAP. Five pseudolesions and 4 hemoangiomas were found in multi-phasic CTAP and CTHA. No pseudolesions and hemoangiomas were shown as rim enhancement on late- or delayed-phasic CTHA and CTAP. CONCLUSION: Multi-phasic CTAP and CTHA could help to recognize the false-positive findings in CTAP and CTHA images, and improve the accuracy of CTAP and CTHA of liver cancer detection.

Adult↗

PubMed2004

Molecular and biochemical characterization of the Skp2-Cks1 binding interface.

SCF(Skp2) is a multisubunit E3 ubiquitin ligase responsible for ubiquitination of cell cycle inhibitor p27. Ubiquitination of p27 requires an adapter protein, Cks1, to be in direct association with Skp2. The exact interface between Skp2 and Cks1 has not been elucidated. Here we have reported the definition of the critical functional interface between Skp2 and Cks1. We have identified eight amino acid residues in two discrete regions of Skp2 that are engaged in Cks1 binding. Mutation of any of these eight residues alone or in combination results in the loss of Cks1 association and negates Skp2-dependent p27 ubiquitination. These eight amino acid residues map on the same side of the Skp2 structure and likely constitute a functional binding surface for Cks1. Four of the eight amino acid residues are located in the largely unstructured carboxyl-terminal tail region of Skp2. These results uncovered the specificity of the Skp2-Cks1 interaction and reveal a critical function for the structurally flexible carboxyl-terminal tail region of Skp2 in Cks1 recognition and substrate ubiquitination.

Amino Acid Motifs↗

PubMed2004

Taurine activates strychnine-sensitive glycine receptors in neurons of the rat inferior colliculus.

Taurine (Tau) is one of the most abundant free amino acids in the mammalian central nervous system. Whether the neurotransmission of the central auditory system is regulated or modulated by Tau is not clear. In the present study, we investigated the electrophysiological and pharmacological properties of Tau-activated currents in acutely dissociated neurons of the rat inferior colliculus (IC) using whole cell patch clamp recordings. At a holding potential of -60 mV and under a condition of chloride equilibrium potential near 0 mV, Tau activated an inward current and its half-maximal activation concentration was equal to 0.37 mM. The measured reversal potential of Tau-activated currents was close to theoretical chloride equilibrium potential. The currents evoked by Tau at both low (1 mM) and high (10 mM) concentrations were almost completely inhibited by strychnine, a glycine receptor antagonist. The Tau-activated current, however, was not affected by bicuculline, a GABA(A) receptor antagonist. Tau at increased concentrations progressively reduced the current response to subsequent glycine application. At saturated concentrations, Tau-activated current and glycine-activated current were mutually cross-desensitized by each other. These findings indicate that Tau activates glycine receptors in neurons of the rat IC and thus may have a functional role in regulating or modulating the neurotransmission of the central auditory system in mammals.

Animals↗

PubMed2004

Structural, biochemical, and functional analyses of CED-9 recognition by the proapoptotic proteins EGL-1 and CED-4.

Programmed cell death in Caenorhabditis elegans is initiated by the binding of EGL-1 to CED-9, which disrupts the CED-4/CED-9 complex and allows CED-4 to activate the cell-killing caspase CED-3. Here we demonstrate that the C-terminal half of EGL-1 is necessary and sufficient for binding to CED-9 and for killing cells. Structure of the EGL-1/CED-9 complex revealed that EGL-1 adopts an extended alpha-helical conformation and induces substantial structural rearrangements in CED-9 upon binding. EGL-1 interface mutants failed to bind to CED-9 or to release CED-4 from the CED-4/CED-9 complex, and were unable to induce cell death in vivo. A surface patch on CED-9, different from that required for binding to EGL-1, was identified to be responsible for binding to CED-4. These data suggest a working mechanism for the release of CED-4 from the CED-4/CED-9 complex upon EGL-1 binding and provide a mechanistic framework for understanding apoptosis activation in C. elegans.

Amino Acid Sequence↗

PubMed2004

Disruption of ceruloplasmin and hephaestin in mice causes retinal iron overload and retinal degeneration with features of age-related macular degeneration.

Mechanisms of brain and retinal iron homeostasis have become subjects of increased interest after the discovery of elevated iron levels in brains of patients with Alzheimer's disease and retinas of patients with age-related macular degeneration. To determine whether the ferroxidase ceruloplasmin (Cp) and its homolog hephaestin (Heph) are important for retinal iron homeostasis, we studied retinas from mice deficient in Cp and/or Heph. In normal mice, Cp and Heph localize to Müller glia and retinal pigment epithelium, a blood-brain barrier. Mice deficient in both Cp and Heph, but not each individually, had a striking, age-dependent increase in retinal pigment epithelium and retinal iron. The iron storage protein ferritin was also increased in Cp-/-Heph-/Y retinas. After retinal iron levels had increased, Cp-/-Heph-/Y mice had age-dependent retinal pigment epithelium hypertrophy, hyperplasia and death, photoreceptor degeneration, and subretinal neovascularization, providing a model of some features of the human retinal diseases aceruloplasminemia and age-related macular degeneration. This pathology indicates that Cp and Heph are critical for CNS iron homeostasis and that loss of Cp and Heph in the mouse leads to age-dependent retinal neurodegeneration, providing a model that can be used to test the therapeutic efficacy of iron chelators and antiangiogenic agents.

Animals↗

PubMed2004

Kinase peptide specificity: improved determination and relevance to protein phosphorylation.

Specificity of phosphorylation is critical to signal transduction. Recent emphasis on colocalization of substrate and kinase has eclipsed emphasis on peptide specificity, i.e., kinase preference for particular amino acids surrounding the phosphorylation site. We describe an approach to determining peptide specificity by using positional scanning of biotinylated oriented peptide libraries and insights emerging from those determinations. We accurately determine preference (or disfavor) for residues at a given substrate position (such as P+2) by comparison of in vitro phosphorylation of peptide libraries differing by a single residue at that position. By analysis of all positions near the phosphorylation site, position-specific scoring matrices are generated and used both to understand the basis of specificity and to predict phosphorylation. PKC-delta and -zeta predictions have been validated rigorously by comparisons with measured phosphorylation. The results demonstrate specificity and sensitivity (80-90%) much better than the previous predictive method. These predictions can be accessed at http://mpr.nci.nih.gov. The accuracy of the specificity determination allows identification of an important difference in peptide specificity between these closely related kinases; Ile/Leu at the P-1 position is disfavored by PKC-zeta but not PKC-delta. Our findings and visual representation of peptide specificity highlight the importance of disfavored residues. Finally, analysis of 124 experimentally determined PKC sites from the literature demonstrates a very strong role of peptide specificity in many of those sites. Thus, position-specific scoring matrices generated by this method provide a foundation for quantitative analyses of kinase specificity and improved predictions of previously determined physiologically relevant phosphorylation sites.

Cyclic AMP-Dependent Protein Kinases↗

PubMed2004

The use of trastuzumab in the treatment of hormone refractory prostate cancer; phase II trial.

PURPOSE: To investigate the efficacy and toxicity of the antibody to the HER-2/neu receptor (trastuzumab, Herceptin) in the treatment of advanced hormone-refractory prostate cancer (HRPC). MATERIALS AND METHODS: Eighteen patients with HRPC were recruited for this phase II trial in which they received trastuzumab for 12 weeks or until disease progression or unacceptable toxicity was documented. HER-2 receptor overexpression was evaluated using immunohistochemistry (IHC) and dual-color fluorescence in-situ hybridization (FISH) assays. RESULTS: Trastuzumab as a single agent demonstrated little efficacy in treating HRPC. Two patients demonstrated stable disease based on a decrease in PSA level to less than 50% of baseline. No patient demonstrated a regression of radiographic bony or soft tissue metastatic disease. The medication was well tolerated in 16 patients (89%), and 2 patients (11%) had to be hospitalized for cardiac complications. CONCLUSIONS: Trastuzumab (Herceptin) as a single agent demonstrated poor efficacy in treating HRPC. Based on promising results in treating breast cancer with regimens using Herceptin and cytotoxic agents, a similar combination approach might demonstrate better efficacy in treating HRPC.

Antibodies, Monoclonal↗

PubMed2004

[Radical resection with total mesorectal excision under the laparoscope in the treatment of rectal cancer].

OBJECTIVE: To evaluate the effect of radical resection with total mesorectal excision under the laparoscope in the treatment of rectal cancer. METHODS: Under the laparoscope and with the ultrasonic scalpel, total mesorectal excision in 38 patients with rectal cancer was performed. In operation, alone the parietal layer of pelvic fascia and inside the autonomic nerve trunk, the mesorectum was excised. All patients were followed up for 2 years. RESULTS: All cases were underwent successfully and no complication was observed. Postoperative flatus and feces were passed at 32 hours and postoperational hospital stay were 7.5 days. No trocar port implantation and no death case was observed. Two local recurrence developed in 2 years follow-up. CONCLUSIONS: Radical resection with TME is a new progress of less damageable operation, which has advantages of low postoperative local recurrence rate and help for pelvic autonomic nerve.

Adult↗

PubMed2004

Secondary Philadelphia chromosome in a patient with acute lymphoblastic leukemia.

We report the case of a patient with acute lymphoblastic leukemia who developed a secondary Philadelphia chromosome-positive clone (Ph(+)). Although the Ph was not detected at diagnosis with conventional cytogenetic analysis and with molecular methods (reverse transcriptase polymerase chain reaction), the Ph was detected with the same techniques at relapse. This anomaly is usually related to the initiation of the disease, but can be a late event related to progression. We discuss these hypotheses and review the literature.

Adult↗

PubMed2004

[The maintenance of cord blood CD34+ progenitor cells with plant lectin FRIL in vitro and the expression of related cell cycle modulator HTm4 and HTm4S].

Ex vivo maintainance of human stem cells is crucial for many clinical applications. Current culture conditions provide some level support but cytokines induce most quiescent stem cells to proliferate and differentiate. Better control of primitive cells is needed to extend the time and range of manipulation of such cells. A recently identified plant lectin Flt3 receptor-interacting lectin (FRIL) present may a special ability to preserve primitive CB progenitors for extended periods in culture without exogenous cytokines. But the mechanisms of FRIL preserving quiescent primitive cells are still unknown. Recently a novel protein HTm4 and its alternatively spliced variant HTm4S, which serve as hematopoietic cell cycle regulators, have been identified. In this report we studied the effect of FRIL on the in vitro maintenance of quiescent human cord blood stem cells and the expression of the novel hematopoietic cell cycle regulator HTm4 and HTm4S in progenitor cells cultured in FRIL. We analyzed the proliferation and the HPP-CFC proportion of CD34(+) cells treated with FRIL. The human HTm4 and HTm4S mRNA expression was detected by semi-quantitative RT-PCR, and the cell cycle status of CB CD34(+) cells was analyzed by FACS. The results showed that incubation of CD34(+) cells in FRIL resulted in a low proliferation of progenitor cells and fewer cycling cells, but FRIL selectively maintained a higher number of primitive cells with proliferative potential in suspension culture. CB CD34(+) cells cultured in FRIL showed significant diversity in the expression of HTm4 and HTm4S during 0~14 d. On d 0, HTm4 was detected at high level, downregulated on d 1, but upregulated during d 3 to d 14, and reaching the highest level on d 7. But the expression levels of HTm4S changed little in the cells cultured in FRIL except the obviously increased expression on d 7. Exogenous expression showed that HTm4 was localized around the karyon while HTm4S scatted in the cytoplasm, respectively, which may be responsible for their difference in function. Thus, FRIL can preserve quiescent primitive CD34(+), and FRIL's ability to preserve quiescent primitive cells in a reversible manner may significantly expand the time and range of ex vivo manipulations of human stem cells for clinical applications. In other words, HTm4 and HTm4S may play a crucial role in the cell cycle modulation of CD34(+) progenitor cells maintained with FRIL in vitro.

Antigens, CD20↗

PubMed2004

Helicobacter pylori L-form and patients with chronic gastritis.

AIM: To study the relationship between infection with Helicobacter pylori L-forms and chronic gastritis and its association with possible changes of cellular immune function. METHODS: Gastric mucosal biopsies were taken from 428 patients with chronic gastritis to detect H pylori L-form by Gram staining and immunohistochemistry staining. Peripheral venous blood samples of patients were taken to detect the percentage of CD3+, CD4+ and CD8+ by the biotin-streptavidin (BSA) assay and the levels of IL-2, IL-6 and IL-8 by ELISA. RESULTS: The rate of infection with H pylori L-forms was 48.83% (209/428). The rate was 50.47% (216/428) and 52.80% (226/428), respectively, as detected by immunohistochemistry staining and Gram staining (P>0.05). The rate of H pylori L-forms in males and females was 57.8% (136/235) and 37.28% (73/193), respectively, (chi2=17.05, P<0.01). Furthermore, the rate increased with age, with the rate being significantly greater in patients > or =40 years old than in those <40 years old (P<0.01). The percentage of CD3+, CD4+, CD8+, the ratio of CD4+/CD8+, and the levels of IL-2, IL-6, IL-8 in H pylori-positive patients were 47.58+/-4.44%, 25.51+/-4.74%, 22.77+/-7.46%, 1.44+/-0.51%, 1.56+/-0.47 mg/L, 103.62+/-5.85 ng/L, and 109.79+/-7.18 ng/L, respectively. Compared with H pylori-negative patients, the percentage of CD3+, CD4+ and the ratio of CD4+/CD8+ and the IL-2 level decreased, but the levels of IL-6, IL-8 increased (P<0.001-P<0.01). Moreover, the percentage of CD3+, CD4+, CD8+, the ratio of CD4+/CD8+, and the levels of IL-2, IL-6 and IL-8 in patients infected with both H pylori L-forms and vegetative forms were 46.67+/-5.21%, 30.75+/-4.89%, 22.15+/-6.45%, 1.32+/-0.47%, 1.16+/-0.38 mg/L, 116.45+/-5.44 ng/L, and 118.64+/-6.24 ng/L, respectively. Compared with patients infected with only vegetative forms, the percentage of CD4+, the ratio of CD4+/CD8+and IL-2 level decreased, but the levels of IL-6 and IL-8 increased (P<0.001 or P<0.01). CONCLUSION: L-form variation often occurs in patients with chronic gastritis and is commonly found in male patients and associates with ages. The L-form variation may be an important factor causing disorder of cellular immune function in the patients with H pylori-induced chronic gastritis.

Adolescent↗

PubMed2004

Docking motif interactions in MAP kinases revealed by hydrogen exchange mass spectrometry.

Protein interactions between MAP kinases and substrates, activators, and scaffolding proteins are regulated by docking site motifs, one containing basic residues proximal to Leu-X-Leu (DEJL) and a second containing Phe-X-Phe (DEF). Hydrogen exchange mass spectrometry was used to identify regions in MAP kinases protected from solvent by docking motif interactions. Protection by DEJL peptide binding was observed in loops spanning beta7-beta8 and alphaD-alphaE in p38alpha and ERK2. In contrast, protection by DEF binding to ERK2 revealed a distinct hydrophobic pocket for Phe-X-Phe binding formed between the P+1 site, alphaF helix, and the MAP kinase insert. In inactive ERK2, this pocket is occluded by intramolecular interactions with residues in the activation lip. In vitro assays confirm the dependence of Elk1 and nucleoporin binding on ERK2 phosphorylation, and provide a structural basis for preferential involvement of active ERK in substrate binding and nuclear pore protein interactions.

Amino Acid Sequence↗

PubMed2004

Proexosite-1-dependent recognition and activation of prothrombin by taipan venom.

An activator complex from the venom of Oxyuranus scutellatus scutellatus (taipan venom) is known to rapidly activate prothrombin to thrombin. To determine whether, similar to prothrombinase, taipan venom utilizes proexosite-1 on prothrombin for a productive complex assembly, the activation of proexosite-1 mutants of prethrombin-1 by the partially purified venom was studied. It was discovered that basic residues of this site (Arg(35), Lys(36), Arg(67), Lys(70), Arg(73), Arg(75), and Arg(77)) are also crucial for recognition and rapid activation of the substrate by taipan venom. This was evidenced by the observation that the K(m) and k(cat) values for the activation of the charge reversal mutants of prethrombin-1 (in particular K36E, R67E, and K70E) were markedly impaired. Competitive kinetic studies with the Tyr(63)-sulfated hirudin(54-65) peptide revealed that although the peptide inhibits the activation of the wild type zymogen by taipan venom with a K(D) of approximately 2 microm, it is ineffective in inhibiting the activation of mutant zymogens (K(D) > 4-30 microm). Interestingly, an approximately 50-kDa activator, isolated from the taipan venom complex, catalyzed the activation of prothrombin in a factor Va-dependent manner and exhibited identical activation kinetics toward the substrate in the presence of the hirudin peptide. These results suggest that, similar to prothrombinase, proexosite-1 is a cofactor-dependent recognition site for taipan venom.

Antithrombins↗

PubMed2004

Ultraviolet sealing and poly(dimethylacrylamide) modification for poly(dimethylsiloxane)/glass microchips.

Simple sealing methods for poly(dimethylsiloxane) (PDMS)/glass-based capillary electrophoresis (CE) microchips by UV irradiation are described. Further, we examined the possibility to modify the inner surface of separation channels, using polymethylacrylamide (PDMA) as a dynamic coating reagent. The surface properties of native PDMS, UV-irradiated PDMS, and PDMA-coated PDMS were systematically studied by atomic force microscopy (AFM), infrared absorption by attenuated total reflection infrared (ATR-IR) spectroscopy, and contact angle measurement. We found that PDMA forms a stable coating on PDMS and glass surfaces, eliminating the nonhomogeneous electroosmotic flow (EOF) in channels on PDMS/glass microchips, and improving the hydrophilicity of PDMS surfaces. Mixtures of flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), and fluorescein were separated in 35 s using PDMA-coated PDMS/glass microchips. A high efficiency of theoretical plates with at least 1365 (105 000 N/m) and a good reproducibility with relative standard deviations (RSD) below 4% in five successive separations were achieved.

Acrylamides↗

PubMed2004

Transfer of brachialis branch of musculocutaneous nerve for finger flexion: anatomic study and case report.

Based on an anatomic study, a transfer of the brachialis muscle branch of the musculocutaneous nerve (BMBMCN) to finger flexor functional fascicles of the median nerve was designed. Preliminary results of clinical application of this new procedure are reported. Dissection of 32 cadaver upper limbs revealed that BMBMCN derives from the musculocutaneous nerve at the distal 1/3 upper arm level. Mostly it is of single-branch type, with an average dissectable length of 5.2 cm. At this level, functional fascicles of finger flexors are located at the posterior 1/3 of the median nerve. BMBMCN can be directly coapted to these finger flexion fascicles. In one case of brachial plexus lower trunk injury, this neurotization procedure was done. No impairment of elbow flexion and wrist flexion was found postoperatively. Recovery of finger and thumb flexion was seen 1 year postoperatively. This neurotization is safe and effective for treating lower trunk injuries.

Adult↗

PubMed2004

Epidermal nerve fiber density, axonal swellings and QST as predictors of HIV distal sensory neuropathy.

We investigated the associations of baseline epidermal nerve fiber (ENF) densities and morphology (percent ENF swellings) and quantitative sensory testing (QST) with clinically defined human immunodeficiency virus (HIV)-associated distal polyneuropathy (DSP) and whether these measures are predictive of development of symptomatic DSP over time. Fifty-seven HIV-infected subjects with and without DSP and 19 controls participated. Mean ENF densities were lower at the distal leg and proximal thigh in asymptomatic or symptomatic DSP than in controls. Mean ENF densities did not differ significantly among the HIV groups. Percent ENF swellings was higher in patients with symptomatic DSP than controls at the distal leg, and was also greater at the proximal thigh in patients with asymptomatic or symptomatic DSP than in controls. The percent ENF swellings at the distal leg correlated with the thresholds for both minimal (HP 0.5) and intermediate (HP 5.0) heat pain (HP) intensity. A higher percent ENF swellings in the distal leg [hazard ratio (HR) 1.16, 95% CI 1.02-1.31] and HP 0.5 thresholds (HR 1.03, 95% CI 1.01-1.05) were the only two measures associated with a shorter time to development of symptomatic DSP. Quantitation of ENF swellings and heat pain thresholds deserve further study as predictors of symptomatic neuropathy.

Adult↗