PubMed Health⌕ Search

Biomedical subjects

Fang Li

Publications and source records attributed to Fang Li.

At least 91 records · Page 5Linked to original sources

[Determination of four nucleosides in Banlangen injections using capillary zone electrophoresis].

The contents of cytidine, adenosine, guanosine and uridine in Banlangen Injections were determined by capillary zone electrophoresis. Shorter fused silica capillary (32.5 cm x 50 microm i. d. with effective length of 23.5 cm) was used. The samples were analyzed with 60 mmol/L borate-10% (v/v) 2-propanol-20% (v/v) acetonitrile running buffer at 20 kV voltage (25 degrees C capillary temperature). Pressure injection of 1 kPa x 10 s was employed, and the detection wavelength was 254 nm. Factors of the electrophoresis condition were investigated, such as the kinds and concentration of the electrolyte, buffer pH value, the type and concentration of organic modifier, separation voltage and temperature. The samples were filtered through 0.45 microm membrane, then direct injection was employed. External calibration of peak area versus concentration was used in the determination. Linear calibrations of four nucleosides were obtained within 12.5 - 250 mg/L (r > 0.999 1). The limits of detection (mg/L) were 6.2 of cytidine, 4.6 of adenosine, 6.7 of guanosine, and 9.0 of uridin. The average recoveries of the four nucleosides were between 95.1% - 102.3% with RSDs of 0.3% - 4.9%. The method is simple, rapid, reproducible and accurate. It can be used for the routine analysis of the four nucleosides in Banlangen Injections.

Adenosine↗

[Analysis of Pb(II), Cd(II) and Cu(II) in Chinese medicine by the system of porphyrin complexes and sulfhydryl cotton fiber].

The reaction of alpha beta gamma delta-tetra(p-sulfophenyl)porphyrin (TPPS4) with Pb(II), Cd(II) or Cu(II) has been studied in this article, and the spectra of the Pb(II)-TPPS4, Cd(II)-TPPS4 and Cu(II)-TPPS4 show the spectral absorption of these complexes with high sensitivity. The molar absorptivities of Pb(II)-TPPS4, Cd(II)-TPPS4 and Cu(II)-TPPS4 are 2.5 x 10(5) L x mol(-1) x cm(-1), 5.2 x 10(5) L x mol(-1) x cm(-1) and 4.2 x 10(5) L x mol(-1) x cm(-1), respectively. With the sulfhydryl cotton fiber separation-enrichment method, this analytical system of porphyrin complexes has been successfully applied to determining the trace amounts of Pb(II), Cd(II) and Cu(II) in Ginkgo bilobal leaves and tea leaves. The RSD of determining 10(-6)-10(-7) g x g(-1) Pb(II), Cd(II) or Cu(II) in samples lies between 3.3%-9.6%, and the recovery of added standard lies between 90%-103%. The proposed analytical method has the advantage of high sensitivity, simplicity and high efficiency of interfere-resisting.

Adsorption↗

Modulation of endothelial Ca(2+)-activated K(+) channels by oxidized LDL and its contribution to endothelial proliferation.

OBJECTIVE: Oxidized low-density lipoprotein (oxLDL) plays an important role in causing endothelial dysfunction and initiating atherosclerosis. Some of the endothelial functions have been shown to be modulated by changes in cellular electrophysiological properties. Therefore, we analysed the effect of oxLDL on endothelial Ca(2+)-activated K(+) channels (BK(Ca)) and its contribution to oxLDL-mediated changes of proliferation and syntheses of nitric oxide (NO). METHODS: The patch-clamp technique was used to study the behavior of BK(Ca) in human endothelial cells of umbilical cord veins (HUVEC). Changes of intracellular Ca(2+) were measured by means of Fura-2 imaging. Cell counts and [3H]-thymidine incorporation were used to analyse endothelial proliferation. Synthesis of NO was measured by means of [3H]-cGMP radioimmunoassay. RESULTS: oxLDL (10 microg/ml) caused a significant increase of BK(Ca) activity, whereas preincubation of HUVEC with an antibody against the lectin-like-oxLDL-receptor-1 (LOX-1) abolished BK(Ca) activation. Fura-2 measurements revealed a biphasic increase of intracellular Ca(2+) after application of the atherogenic lipid. Endothelial proliferation was significantly increased by oxLDL. The highly selective BK(Ca) inhibitor iberiotoxin (100 nmol/l IBX) blocked this proliferative response. Acetylcholine-induced NO synthesis was significantly decreased by IBX. Interestingly, oxLDL significantly decreased acetylcholine-induced NO synthesis if the production of superoxide was not blocked by antisense oligonucleotides against the NAD(P)H-oxidase. CONCLUSIONS: Our data demonstrate that oxLDL activates BK(Ca), which plays an important role in oxLDL-mediated endothelial proliferation. Acetylcholine-induced NO synthesis is modulated by BK(Ca), whereas the reduction of acetylcholine-induced NO-synthesis by oxLDL is related to an increase in superoxide production.

Acetylcholine↗

A novel method of determining the number of macromolecules per asymmetric unit from accurate crystal-volume measurements.

Knowledge of the number of macromolecules per crystallographic asymmetric unit is frequently useful in the determination of crystal structures. A method has been developed to establish this number directly from measurements of the volume and macromolecular contents of a crystal. The volume of a crystal is determined by measuring the volume of solvent that it displaces in a fine capillary tube. The macromolecular mass contained in a crystal is measured by dissolving the crystal in a known amount of water or suitable buffer and then measuring the UV absorbance of the solution. The method has been tested successfully on three different crystals of known structures.

Animals↗

The STT3a subunit isoform of the Arabidopsis oligosaccharyltransferase controls adaptive responses to salt/osmotic stress.

Arabidopsis stt3a-1 and stt3a-2 mutations cause NaCl/osmotic sensitivity that is characterized by reduced cell division in the root meristem. Sequence comparison of the STT3a gene identified a yeast ortholog, STT3, which encodes an essential subunit of the oligosaccharyltransferase complex that is involved in protein N-glycosylation. NaCl induces the unfolded protein response in the endoplasmic reticulum (ER) and cell cycle arrest in root tip cells of stt3a seedlings, as determined by expression profiling of ER stress-responsive chaperone (BiP-GUS) and cell division (CycB1;1-GUS) genes, respectively. Together, these results indicate that plant salt stress adaptation involves ER stress signal regulation of cell cycle progression. Interestingly, a mutation (stt3b-1) in another Arabidopsis STT3 isogene (STT3b) does not cause NaCl sensitivity. However, the stt3a-1 stt3b-1 double mutation is gametophytic lethal. Apparently, STT3a and STT3b have overlapping and essential functions in plant growth and developmental processes, but the pivotal and specific protein glycosylation that is a necessary for recovery from the unfolded protein response and for cell cycle progression during salt/osmotic stress recovery is associated uniquely with the function of the STT3a isoform.

Adaptation, Physiological↗

The mouse Formin mDia1 is a potent actin nucleation factor regulated by autoinhibition.

Formin proteins are widely expressed in eukaryotes and play essential roles in assembling specific cellular actin-based structures. Formins are defined by a Formin Homology 2 (FH2) domain, as well as a proline-rich FH1 domain that binds the actin monomer binding protein, profilin, and other ligands. Constructs including FH2 of budding yeast Bni1 or fission yeast Cdc12 formins nucleate actin filaments in vitro. In this study, we demonstrate that FH2-containing constructs of murine mDia1 (also called p140 mDia or Drf1) are much more potent actin nucleators than the yeast formins. FH1 is necessary for nucleation when actin monomers are profilin bound. mDia1 is a member of the Diaphanous formin subfamily (Dia), whose members contain an N-terminal Rho GTPase binding domain (GBD) and a C-terminal Diaphanous autoinhibitory domain (DAD, ). Based on cellular and in vitro binding studies, an autoinhibitory model for Dia formin regulation proposes that GBD binding to DAD inhibits Dia-induced actin remodeling, whereas Rho binding activates by releasing GBD from DAD. Supporting this model, our results show that an N-terminal mDia1 construct strongly inhibits actin nucleation by the C terminus. RhoA partially relieves inhibition but does so when bound to either GDP or GTP analogs. Both N- and C-terminal mDia1 constructs appear to be multimeric.

Actins↗

Prestin, a cochlear motor protein, is defective in non-syndromic hearing loss.

Prestin, a membrane protein that is highly and almost exclusively expressed in the outer hair cells (OHCs) of the cochlea, is a motor protein which senses membrane potential and drives rapid length changes in OHCs. Surprisingly, prestin is a member of a gene family, solute carrier (SLC) family 26, that encodes anion transporters and related proteins. Of nine known human genes in this family, three (SLC26A2, SLC26A3 and SLC26A4) are associated with different human hereditary diseases. The restricted expression of prestin in OHCs, and its proposed function as a mechanical amplifier, make it a strong candidate gene for human deafness. Here we report the cloning and characterization of four splicing isoforms for the human prestin gene (SLC26A5a, b, c and d). SLC26A5a is the predominant form of prestin whereas the others showed limited distribution associated with certain developmental stages. Based on the functional importance of prestin we screened for possible mutations involving the prestin gene in a group of deaf probands. We have identified a 5'-UTR splice acceptor mutation (IVS2-2A>G) in exon 3 of the prestin gene, which is responsible for recessive non-syndromic deafness in two unrelated families. In addition, a high frequency of heterozygosity for the same mutation was observed in these subjects, suggesting the possibility of semi-dominant influence of the mutation in causing hearing loss. Finally, the observation of this mutation only in the Caucasian probands indicated an association with a specific ethnic background. This study thereby reveals an essential function of prestin in human auditory processing.

Alternative Splicing↗

Development-associated myristoylated alanine-rich C kinase substrate phosphorylation in rat brain.

OBJECT: In neuronal cells, myristoylated alanine-rich C kinase substrate (MARCKS), localized to particular areas of the synaptic membrane, is active during brain development. The destination of phosphorylated MARCKS is thought to be the cytoplasm where it is probably inactive. We compared MARCKS phosphorylation in the brains of embryonic, perinatal, and adult rats to determine its possible involvement in neurogenesis. METHODS: We prepared crude and partially purified extracts from various brain regions of rats aged between embryonic day 14 (E14) and 7 weeks after birth and assayed them for MARCKS phosphorylation by immunochemical methods. The isotypes of protein kinase C (PKC) were immunochemically identified in crude brain extracts from embryonic and postnatal rats. Despite negligible MARCKS phosphorylation, E16 brain extracts contained both MARCKS and PKCgamma, delta, epsilon, and lambda. MARCKS and polypeptides were clearly phosphorylated (49 and 45 kDa, respectively) in brain extracts purified on a DE52 column. Embryonic brain extracts manifested a high-molecular-weight activity capable of suppressing polypeptide phosphorylation. This activity was markedly decreased on the day of birth and almost undetectable in the brains of 9-day-old rats. CONCLUSIONS: The embryonic rat brain appears to contain a protein(s) that suppresses the phosphorylation of other proteins including MARCKS. We posit that this inhibitory activity represents a factor(s) that plays a role in the regulation of neurogenesis beginning on the day on which MARCKS appears in the embryonic brain.

Animals↗

Relationship between apoptosis and E-cadherin expression in bronchial epithelium of smoking mouse.

To investigate whether apoptosis is associated with cell adhesion in bronchial epithelium, and whether it contributes to the kinetics of injury and repair of surface epithelia, this study was performed for E-cadherin expression by using immunohistochemistry technique and for apoptosis by TUNEL method. An animal model of smoking was used for this study. The results showed that epithelial cells with membrane anchored E-cadherin decreased remarkably at several time points during 6 months of exposure to smoke (P < 0.01) and then restored to normal level. This fluctuation was associated exclusively with the alteration in number of apoptotic cells (P < 0.01). There was no significant difference in activation of nuclear transcription factor NF-kappa B among groups (P > 0.05). All these suggested that apoptosis is associated with E-cadherin expression in bronchial epithelium of smoking mouse.

Animals↗

Crystal structures of an archaeal class I CCA-adding enzyme and its nucleotide complexes.

CCA-adding enzymes catalyze the addition of CCA onto the 3' terminus of immature tRNAs without using a nucleic acid template and have been divided into two classes based on their amino acid sequences. We have determined the crystal structures of a class I CCA-adding enzyme from Archeoglobus fulgidus (AfCCA) and its complexes with ATP, CTP, or UTP. Although it and the class II bacterial Bacillus stearothermophilus CCA enzyme (BstCCA) have similar dimensions and domain architectures (head, neck, body, and tail), only the polymerase domain is structurally homologous. Moreover, the relative orientation of the head domain with respect to the body and tail domains, which appear likely to bind tRNA, differs significantly between the two enzyme classes. Unlike the class II BstCCA, this enzyme binds nucleotides nonspecifically in the absence of bound tRNA. The shape and electrostatic charge distribution of the AfCCA enzyme suggests a model for tRNA binding that accounts for the phosphates that are protected from chemical modification by tRNA binding to AfCCA. The structures of the AfCCA enzyme and the eukaryotic poly(A) polymerase are very similar, implying a close evolutionary relationship between them.

Adenosine Triphosphate↗

Tumour necrosis factor-alpha (TNF-alpha) transgene-expressing dendritic cells (DCs) undergo augmented cellular maturation and induce more robust T-cell activation and anti-tumour immunity than DCs generated in recombinant TNF-alpha.

Tumour antigen presentation by dendritic cells (DCs) to T cells in lymphoid organs is crucial for induction of anti-tumour immune responses. It has been previously reported that tumour necrosis factor-alpha (TNF-alpha) is required for DC activation and subsequent induction of optimal immune responses, and thus DCs for anti-tumour vaccination are often generated by culture in exogenous TNF-alpha. In the present study, we investigated the effect on anti-tumour immunity of vaccination with Mut1 tumour peptide-pulsed DCs engineered to express a TNF-alpha transgene. Our data shows that transfection of DCs with recombinant adenovirus AdV-TNF-alpha resulted in greater maturation of the DCs than occurred with control DCs cultured in exogenous TNF-alpha, as determined by up-regulated expression of pro-inflammatory cytokines (e.g. interleukins 1beta and 18), chemokines [e.g. interferon-gamma-inducible protein-10 and macrophage inflammatory protein-1beta (MIP-1beta)], the CC chemokine receptor CCR7, and immunologically important cell surface molecules (CD40, CD86 and intercellular adhesion molecule-1). These transgenic DCs stimulated stronger allogeneic T-cell responses in vitro and T-cell activation in vivo; displayed 2.4-fold enhanced chemotactic responses to the MIP-3betain vitro (P<0.05); and, perhaps most importantly, trafficked into the draining lymph nodes dramatically (seven-fold, P<0.01) more efficiently than the control DCs. Our data also demonstrate that vaccination of mice with Mut1 peptide-pulsed, AdV-TNF-alpha-transfected DCs stimulated more efficient in vitro Mut1-specific CD8+ cytotoxic T-cell responses and solid tumour immunity in vivo, when compared to the in vitro TNF-alpha-cultivated DCs. Thus, DCs engineered to secrete TNF-alpha may offer a new strategy in DC cancer vaccines.

Adenoviridae↗

Oligoclonal expansions of antigen-specific CD8+ T cells in aged mice.

Oligoclonal T cell expansions (TCE) are common in old humans and mice. It is not known whether an Ag-specific response becomes more oligoclonal with age, and, if so, how this might alter biological responses or compromise the immune response, thus contributing to the immunodeficiency of aging. We used a tumor antigen response to study these questions. Early on, antigen reactive T cell numbers at the site of tumor injection were lower and clonally more restricted in old mice. Subsequently, long-term oligoclonal TCE emerged in the blood and spleen of old mice. IL-15 was not necessary for development of TCE in the blood. Overall, the data pointed to a dysregulated immune response in old mice, perhaps due to lack of optimal IL-2 and CD4 help at the earliest stages and a lack of an efficient local peritoneal CTL response. This was associated with a deficient humoral response and, likely, persistence of tumor cells or tumor antigens. Perhaps the spleen is the site of persistence which explains clonal TCE observed primarily in PBL and spleen. The TCE appear to be inefficient as they are often anergic. As a result an occasional peritoneal or splenic tumor may arise in old mice.

Aging↗

EAP1, a Candida albicans gene involved in binding human epithelial cells.

Candida albicans adhesion to host tissues contributes to its virulence and adhesion to medical devices permits biofilm formation, but we know relatively little about the molecular mechanisms governing C. albicans adhesion to materials or mammalian cells. Saccharomyces cerevisiae provides an attractive model system for studying adhesion in yeast because of its well-characterized genetics and gene expression systems and the conservation of signal transduction pathways among the yeasts. In this study, we used a parallel plate flow chamber to screen and characterize attachment of a flo8Delta S. cerevisiae strain expressing a C. albicans genomic library to a polystyrene surface. The gene EAP1 was isolated as a putative cell wall adhesin. Sequence analysis of EAP1 shows that it contains a signal peptide, a glycosylphosphatidylinositol anchor site, and possesses homology to many other yeast genes encoding cell wall proteins. In addition to increasing adhesion to polystyrene, heterologous expression of EAP1 in S. cerevisiae and autonomous expression of EAP1 in a C. albicans efg1 homozygous null mutant significantly enhanced attachment to HEK293 kidney epithelial cells. EAP1 expression also restored invasive growth to haploid flo8Delta and flo11Delta strains as well as filamentous growth to diploid flo8/flo8 and flo11/flo11 strains. Transcription of EAP1 in C. albicans is regulated by the transcription factor Efg1p, suggesting that EAP1 expression is activated by the cyclic AMP-dependent protein kinase pathway.

Amino Acid Motifs↗

Human Nudel and NudE as regulators of cytoplasmic dynein in poleward protein transport along the mitotic spindle.

Emerging evidence supports the idea that a signaling pathway containing orthologs of at least mammalian NudE and Nudel, Lis1, and cytoplasmic dynein is conserved for eukaryotic nuclear migration. In mammals, this pathway has profound impact on neuronal migration during development of the central nervous system. Lis1 and dynein are also involved in other cellular functions, such as mitosis. Here we show that Nudel also participates in a subset of dynein function in M phase. Nudel was specifically phosphorylated in M phase in its serine/threonine phosphorylation motifs, probably by Cdc2 and also Erk1 and -2. A fraction of Nudel bound to centrosomes strongly in interphase and localized to mitotic spindles in early M phase. By using mutants incapable of or simulating phosphorylation, we confirmed that phosphorylation of Nudel regulated the cell-cycle-dependent distribution, possibly by increasing its dissociation rate at the microtubule-organizing center. Moreover, phosphorylated Nudel or the phosphorylation-mimicking mutant bound Lis1 more efficiently. We further demonstrated that a Nudel mutant incapable of binding to Lis1 impaired the poleward movement of dynein and hence the dynein-mediated transport of kinetochore proteins to spindle poles along microtubules, a process contributing to inactivation of the spindle checkpoint in mitosis. These results point to the importance of Nudel-Lis1 interaction for the dynein activity in M phase and to a possible role of Nudel phosphorylation as facilitating such interaction. In addition, comparative studies suggest that NudE is also functionally related to its paralog, Nudel.

1-Alkyl-2-acetylglycerophosphocholine Esterase↗

[Assessment of preoperative localization techniques for patients with primary hyperparathyroidism].

OBJECTIVE: To evaluate the sensitivity and usefulness of 99mTc-sestamibi scintigraphy (SS) and neck ultrasonography (US) as preoperative localization procedures in patients with primary hyperparathyroidism (pHPT). METHODS: 160 patients with proved pHPT in Peking Union Medical College Hospital from June 1983 to June 2002 were studied. There were 107 women(66.9%) and 53 men (33.1%), with a mean age of 38.9 years (10-73 years). 100 patients were underwent SS and 148 patients were underwent US prior to surgery, and the results were compared with operative and histological findings. RESULTS: The sensitivity of SS and US in localization of the enlarged parathyroid glands was 94.0% and 85.1% respectively, and the positive predictive value of SS and US was 100% and 89.1% respectively, the overall sensitivity was 98.9% by combination of SS and US. In solitary parathyroid adenomas group (n = 145), the sensitivity of SS and US was 93.3% and 84.7% respectively; There was no significant difference (P = 0.428) in sensitivity of SS between the parathyroid glands correctly identified and undetected in classical neck location as compared with ectopic parathyroid glands, whereas significantly (P = 0.026) influenced by the US sensitivity. CONCLUSIONS: Different sensitivity exit between SS and VS in preoperative localization in patients with pHPT undergoing parathyroidectomy. The combined use of SS and US could increase the sensitivity of localization technique. Ectopic parathyroid had no influence on the sensitivity of 99mTc-MIBI scanning, but decreased the sensitivity of ultrasonography. The size of parathyroid tumors had effects on the sensitivity of ultrasonography. Otherwise, various conditions causing SS false negative were observed. Some interfere factors should be excluded when SS negative results were encountered in clinical practice.

Adolescent↗

[Investigation of concentration of microcystin in raw materials and finished products of spires health food].

In order to investigate the concentration of micromystin in raw materials and finished products of spires health food, we collected 33 water samples from water resources, 160 water samples from culturing pool, 86 samples of Spires slurry and 70 samples of powders of Spires raw materials from 7 Spires manufacturers in Jiangsu, Yunnan, Fujian and Guandong provinces which are the main spires producing areas from July to August 2002. At the same time, we also collected 71 samples of 19 finished products of Spires health foods in markets of above-mentioned 4 provinces. Microcystin were tested for all samples by ELISA. The results show: microcystin were not detected in 12 tap water samples, were detected in 6 of 9 under water samples, 8 of 12 surface water samples; microcystin concentration of water samples from culturing pool was 207.9 pg/ml and Spires slurry was 31.9 ng/g, the former is 153 times as the latter; powder of Spires raw material was 206.4 ng/g. The average concentration of microcystin of all Spires health food samples from market was 317.2 ng/g, there into, microcystin was 142.7 ng/g in a tables and 222.6 ng/g in capsule, respectively. The results suggested: we shouldn't ignore the risk of exposuring microcystin when consuming Spires health food, it is necessary to further study and set a standard of limited microcystin in Spires health food.

Cyanobacteria↗

[Preliminary clinical application of 99Tcm-HYNIC-TOC imaging in somatostatin receptor-positive tumors].

OBJECTIVE: To evaluate the effect of 99Tcm-HYNIC-TOC imaging in localization of somatostatin receptor-positive tumors. METHODS: Forty-four patients were involved in this study, including 22 neuroendocrine tumors, 10 non-neuroendrocrine tumors and 12 benign diseases. All patients were confirmed by histopathologic diagnosis, and had clinical laboratory data, or 1-2 other imaging procedures. Regional, whole body and SPECT/CT (in positive cases) imagings were acquired at 1 and 4 hours after an intravenous injection of 370 MBq 99Tcm-HYNIC-TOC. 99Tcm-HYNIC-TOC imaging was compared with 111In-petetreotide imaging in 4 cases, and with 131I-MIBG imaging in 10 cases. 99Tcm-HYNIC-TOC imaging was performed before and after treatment in 1 non-Hodgkins lymphoma (NHL) patient. RESULTS: The positive imagings were observed in 19 of 32 cases. The sensitivity, specificity, and accuracy of 99Tcm-HYNIC-TOC imaging for somatostatin receptor-positive tumors are 82.6%, 100%, and 87.5%, respectively. The distribution in vivo of 99Tcm-HYNIC-TOC is similar to that of 111In-petetreotide, and showed high physiological uptake in liver, spleen, and kidneys. 99Tcm-HYNIC-TOC imaging demonstrated intense tumor sites uptake at 1 hour after injection, and revealed the lesions first in 6 patients among the imaging modalities, and more lesions that had not been revealed by 131I-MIBG imaging. Compared with imaging before treatment, 99Tcm-HYNIC-TOC imaging confirmed the tumor regression after treatment in 1NHL. CONCLUSIONS: 99Tcm-HYNIC-TOC is promising for the diagnosis and localization of somatostatin receptor-positive tumors.

Adenoma, Chromophobe↗