Biomedical subjects

S Lei

Publications and source records attributed to S Lei.

At least 37 records · Page 2Linked to original sources

PubMed2000

Proteasome inhibition circumvents solid tumor resistance to topoisomerase II-directed drugs.

Physiological cell conditions, such as glucose deprivation and hypoxia, play a role in developing drug resistance in solid tumors. These tumor-specific conditions cause decreased expression of DNA topoisomerase IIalpha (topo IIalpha), rendering cells resistant to topo II-targeted drugs, such as etoposide and doxorubicin. We show here that inhibition of proteasome attenuated drug resistance by inhibiting topo IIalpha depletion induced by glucose starvation and hypoxia. topo IIalpha restoration was seen only at the protein levels, indicating that the topo IIalpha protein depletion occurred through a proteasome-mediated degradation mechanism. The stress-induced etoposide resistance was effectively prevented in vitro by the proteasome inhibitor lactacystin in both intrinsically resistant and sensitive tumor cells (colon cancer HT-29 and ovarian cancer A2780 cells, respectively). Furthermore, lactacystin effectively enhanced the antitumor activity of etoposide in the refractory HT-29 xenograft. These results indicate that lactacystin could serve as a new therapeutic agent to circumvent resistance to topo II-targeted chemotherapy in solid tumors.

Acetylcysteine↗

PubMed2000

Neuronal nicotinic receptors in non-neuronal cells: new mediators of tobacco toxicity?

The nicotinic acetylcholine receptors are prototypic ionotropic receptors that mediate fast synaptic transmission. However, also non-excitable cells, and particularly the tegumental cells that line external and internal body surfaces, express acetylcholine receptors of neuronal type sensitive to nicotine. Bronchial epithelial cells, endothelial cells of blood vessels and skin keratinocytes express neuronal nicotinic receptors composed of alpha(3), alpha(5), beta(2) and beta(4) subunits, similar to those expressed in sympathetic ganglia, and neuronal nicotinic receptors composed of alpha(7) subunits. Neuronal nicotinic receptors in tegumental cells are involved in modulating cell shape and motility, and therefore in maintaining the integrity of the surfaces lined by those cells. Neuronal nicotinic receptors in non-neuronal tissues may modulate other functions, including cell proliferation and differentiation. Acetylcholine is synthesized, secreted and degraded by a variety of cells, including the tegumental cells that express neuronal nicotinic receptors. Thus, acetylcholine may function as a local "hormone" that is able to modulate cell functions that require fast adaptation to new conditions. The presence of neuronal nicotinic receptors sensitive to nicotine in tissues known to be involved in tobacco toxicity, like bronchi and blood vessels, raises the possibility that they mediate some of the toxic effects of smoking.

Bronchi↗

PubMed2000

Cyclic GMP-dependent feedback inhibition of AMPA receptors is independent of PKG.

In central neurons, the second messenger cGMP is believed to induce long-term changes in efficacy at glutamatergic synapses through activation of protein kinase G (PKG). Stimulating nitric oxide synthase, activating soluble guanylyl cyclase or elevating concentrations of intracellular cGMP depressed excitatory synaptic transmission in CA1 hippocampal neurons. Unexpectedly, intracellular cGMP depressed responses of AMPA receptors and inhibited excitatory postsynaptic currents in hippocampal neurons independently of phosphorylation. Our findings demonstrate that cGMP's modulation of excitatory transmission may involve a coupling of AMPA channel activity to levels of cGMP.

Animals↗

PubMed2000

Immunotherapy of tumors with xenogeneic endothelial cells as a vaccine.

The breaking of immune tolerance against autologous angiogenic endothelial cells should be a useful approach for cancer therapy. Here we show that immunotherapy of tumors using fixed xenogeneic whole endothelial cells as a vaccine was effective in affording protection from tumor growth, inducing regression of established tumors and prolonging survival of tumor-bearing mice. Furthermore, autoreactive immunity targeting to microvessels in solid tumors was induced and was probably responsible for the anti-tumor activity. These observations may provide a new vaccine strategy for cancer therapy through the induction of an autoimmune response against the tumor endothelium in a cross-reaction.

Amino Acid Sequence↗

PubMed2000

[Effect of temperature on energy budget of Taiwanese red tilapia hybrid (Oreochromis niloticus x O. mossambicus)].

The energy budget of Taiwanese red tilapia hybrid (Oreochromis niloticus x O. massambicus) with a body weight of 25.33-26.23 g was measured at salinity 14 and at water temperature 22, 28 and 34 degrees C in July-August 1996. The results showed that its specific growth rate, conversion efficiency and maximum food consumption were significantly affected by temperature, and reached peaks of 1.79, 60.01% and 2.59% B.W.d-1 at 28 degrees C. The effect of temperature on the absorption efficiency was not significant, with an average absorption efficiency 64.58%. The effect of temperature on the proportions of food energy allocated to each component of the energy budget was significant. The ratio of growth energy was the highest(26.4%) at temperature 28 degrees C, and that of metabolism energy was 37.23%.

Absorption↗

PubMed2000

Kinetic study of eosinophil apoptosis and its relationship to Th2 cytokines in asthmatic murine model.

OBJECTIVE: To investigate the significance and regulatory mechanism of eosinophil apoptosis in asthmatic airway inflammation using an animal models, as well as its relationship to IL-5 and IL-10. METHODS: An animal model of asthma was established by ovalbumin (OVA) sensitizing-challenging BALB/C mice. At seven points in the time course after challenge, bronchoalveolar lavage (BAL) was performed to obtain BAL cells. Levels of two Th2 cytokines, IL-5 and IL-10 were detected by ELISA. A double label technique combining CD15 with CD49d was used to distinguish eosinophils from other cell types. Cell apoptosis was assayed by PI stain using a flow cytometer. RESULTS: Eosinophil apoptosis was identified and a two-phase elevation of apoptosis percentages was observed, although no significant difference was found between average apoptosis in the asthmatic group and that in the control. Analysis of kinetic changes of eosinophil apoptosis and cytokine levels revealed a negative correlation between apoptosis percentages and levels of IL-5, and a closer correlation between apoptosis percentages and the IL-5/IL-10 ratio. Dexamethasone pretreatment markedly reduced IL-5 levels in BAL, elevated eosinophil apoptosis percentages, but had no effect on IL-10 levels. CONCLUSION: Eosinophils underwent apoptosis in asthmatic airways. Equilibrium between IL-5 and IL-10 not only regulated eosinophil infiltration, but also modulated cell apoptosis.

Animals↗

PubMed2000

[Cell cycle analysis and pathological changes of malignant tumors treated with electrochemical therapy].

To investigate the mechanism and to observe the effectiveness of electrochemical therapy(ECT), 33 patients of late stage cancers which treated by ECT were included in this study. Flow cytometry (FCM) and pathology were used to observe the changes of tumor cells before and after ECT. Tumor cells of G1, G2/M, S phases and aneuploid cells were found to be killed almostly and the ratio of apoptosis was elevated greatly after ECT. Also, pathological evidence proved the death of tumor cells and some characters of apoptosis. So, ECT can non-selectively kill tumor cells and induce the apoptosis of them, and FCM was a way to evaluate the effectiveness of ECT.

Adult↗

PubMed2000

[The relationship of cellular DNA content with clinical stage and biological features of colorectal cancer].

The purpose of this study was to explore the relationship of cellular DNA content with clinical stage and biological features of cancer. Flow cytometry was performed on fresh specimens from 86 patients from 1997 to 1998. Forty-five (53.3%) specimens were found to contain cells with abnormal DNA (DNA nondiploidy). Although none of the sex, age, site, differentiation variables correlated with flow cytometric DNA ploidy, nondiploidy was associated with Dukes' stage and lymph node metastasis. Duke's A stage tumors were more frequently diploid than were more advanced tumors, but no difference between Duke's B, C and D stages were observed. These findings suggest that ploidy is associated with some pathological factors.

Aged↗

PubMed2000

[The relations of cellular DNA content to lymph node metastasis in gastric carcinoma].

The objectives of this study were to evaluate the relations of cellular DNA content, S-phase fraction (SPF) and G2/M-phase(G2/MPF) to lymph node metastasis in gastric carcinoma by using flow cytometry. The results showed that the incidence of lymph node metastasis and SPF were significantly higher in aneuploid carcinoma than in diploid carcinoma(P < 0.05). But the G2/MPF values did not correlate with lymph node metastasis. These findings suggest that correlations between cellular DNA content, SPF and the extent of lymph node metastasis are present.

Adenocarcinoma↗

PubMed2000

[Quantification analysis of vascular endothelial cells in human solid tumor].

Angiogenesis is essential for the growth and metastasis of solid tumors. Intratumoral microvessel count, which represents a measure of tumor angiogenesis, has been associated with prognosis of patients with a variety of malignancies. But till now, it is still a problem to quantitate the angiogenesis of tumors exactly. In this study, the endothelial cells in 20 solid tumors were conjugated with anti-CD34 monoclonal antibodies labelled with red-fluorescent substances, and were observed by fluorescence microscopy and quantitated by flow cytometry. The results of flow cytometry were analysed by t-test. The correlation was analysed between percentage of flow cytometry and vessel counts (light microscopy, x 200) in each tumor which was stained with anti-CD34 monoclonal antibodies and anti-VIII factor related antigen polyclonal antibodies by using immunohistochemistry. The results showed there was significant difference between the treatment group and control group in flow cytometry, and a significant correlation between the endothelial cells percentage by flow cytometry and the vessel counts by immunohistochemistry was observed, although the average vessel counts were approximately three times with CD34 staining as much as with VIII factor related antigen staining. These data indicate that flow cytometry as a novel method can quantitate angiogenesis of tumors exactly and quickly.

Antibodies, Monoclonal↗

PubMed1999

Platelet-derived growth factor receptor-induced feed-forward inhibition of excitatory transmission between hippocampal pyramidal neurons.

Growth factor receptors provide a major mechanism for the activation of the nonreceptor tyrosine kinase c-Src, and this kinase in turn up-regulates the activity of N-methyl-D-aspartate (NMDA) receptors in CA1 hippocampal neurons (1). Unexpectedly, applications of platelet-derived growth factor (PDGF)-BB to cultured and isolated CA1 hippocampal neurons depressed NMDA-evoked currents. The PDGF-induced depression was blocked by a PDGF-selective tyrosine kinase inhibitor, by a selective inhibitor of phospholipase C-gamma, and by blocking the intracellular release of Ca(2+). Inhibitors of cAMP-dependent protein kinase (PKA) also eliminated the PDGF-induced depression, whereas a phosphodiesterase inhibitor enhanced it. The NMDA receptor-mediated component of excitatory synaptic currents was also inhibited by PDGF, and this inhibition was prevented by co-application of a PKA inhibitor. Src inhibitors also prevented this depression. In recordings from inside-out patches, the catalytic fragment of PKA did not itself alter NMDA single channel activity, but it blocked the up-regulation of these channels by a Src activator peptide. Thus, PDGF receptors depress NMDA channels through a Ca(2+)- and PKA-dependent inhibition of their modulation by c-Src.

Animals↗

PubMed1999

Regulated expression of the nifM of Azotobacter vinelandii in response to molybdenum and vanadium supplements in Burk's nitrogen-free growth medium.

Azotobacter is a diazotrophic bacterium that harbors three genetically distinct nitrogenases referred to as nif, vnf, and anf systems. The nifM is an accessory gene located in the nif gene cluster and is transcriptionally regulated by the NifA. However, Azotobacter mutants that lack NifA are known to synthesize functional NifM and this accessory protein is known to be needed for the activity of nitrogenase-2 and nitrogenase-3. To determine how the transcription of nifM is regulated when Azotobacter is grown under conditions in which nitrogenase-2 or nitrogenase-3 is expressed, we generated an Azotobacter vinelandii strain that carries a nifM:lacZ-kanamycin resistance gene cassette in its chromosome. In this strain the nifM open reading frame was disrupted by the presence of a lacZ-kanamycin resistance gene cassette so that it could not produce active NifM. Moreover, the lacZ gene was placed under the transcriptional control elements of the nifM gene so that the lacZ expression could be used as a marker to determine the extent of expression of the nifM gene under different growth conditions. Our results show that this strain was unable to grow in Burk's nitrogen-free medium supplemented with either molybdenum or vanadium or lacking both metals suggesting that in the absence of functional NifM none of the nitrogenases were active. It was also found that the nifM expression was differentially regulated when the A. vinelandii cells were grown under conditions that activate nitrogenase-2 and nitrogenase-3, as determined by liquid beta-galactosidase activity measurements. These results suggest that the transcriptional activators, VnfA and AnfA, may regulate the nifM expression.

Azotobacter vinelandii↗

PubMed1999

Mechanisms involved in the vasorelaxant effect of (-)-stepholidine in rat mesenteric small arteries.

The purpose of the present investigation was to clarify whether the hypotensive action of the protoberberine alkaloid, and dopamine receptor antagonist, (-)-stepholidine, can be ascribed to an effect on peripheral small arteries. For this purpose isolated mesenteric small arteries were suspended in microvascular myographs for isometric tension recording. Relaxations mediated by dopamine D1 receptors were antagonized by (-)-stepholidine. (-)-Stepholidine inhibited in a concentration-dependent manner the contractile responses evoked by noradrenaline (10(-6) M), but not the contractile responses evoked by depolarizing solution (KCl, 60 mM) or 9,11-dideoxy-11alpha,9alpha-epoxymethano prostaglandin F2alpha (U46619, 10(-7) M). Mechanical endothelial cell removal, blockade of K+ channels, muscarinic receptors or adrenoceptors did not influence the inhibitory effect of (-)-stepholidine on the contractile response evoked with noradrenaline in the segments. (-)-Stepholidine caused rightward shifts of the concentration-response curves for noradrenaline and phenylephrine. The pA2 values for (-)-stepholidine were 6.05 and 5.94 against noradrenaline and phenylephrine, respectively. Electrical field stimulation induced prazosin-sensitive frequency-dependent contractions in mesenteric small arteries. These contractions were significantly inhibited by 10(-6) and 10(-5) M (-)-stepholidine. In membranes from the rat cerebral cortex labelled with [3H]prazosin, (-)-stepholidine (10(-7)-10(-4) M) completely inhibited the specific binding of the ligand with a pKi of 5.6. The present investigation suggests the inhibitory effect of (-)-stepholidine on the alpha1-adrenoceptor-mediated contractions induced by exogenously added and nerve-released noradrenaline in peripheral small arteries might contribute to a hypotensive effect of the drug.

Animals↗

PubMed1999

G-protein-coupled receptors act via protein kinase C and Src to regulate NMDA receptors.

The N-methyl-D-aspartate (NMDA) receptor contributes to synaptic plasticity in the central nervous system and is both serine-threonine and tyrosine phosphorylated. In CA1 pyramidal neurons of the hippocampus, activators of protein kinase C (PKC) as well as the G-protein-coupled receptor ligands muscarine and lysophosphatidic acid enhanced NMDA-evoked currents. Unexpectedly, this effect was blocked by inhibitors of tyrosine kinases, including a Src required sequence and an antibody selective for Src itself. In neurons from mice lacking c-Src, PKC-dependent upregulation was absent. Thus, G-protein-coupled receptors can regulate NMDA receptor function indirectly through a PKC-dependent activation of the non-receptor tyrosine kinase (Src) signaling cascade.

Alkaloids↗

PubMed1999

Composition of long chain polyunsaturated fatty acids in human milk and growth of young infants in rural areas of northern China.

The main source of fat in the diet in rural areas of northern China is soybean oil, therefore the pattern of essential fatty acids in human milk may be assumed to differ from that in milk from women in Western countries and to be similar to that of vegans. The concentrations of long chain polyunsaturated fatty acids in human milk and information on diet were analysed for 41 lactating women in rural areas of north China, and the growth of their infants was measured. The subjects were divided into two groups (group I, 1 mo postpartum; group II, 3 mo postpartum). The dietary intake of the mothers was high in carbohydrate and low in fat, protein and energy. The concentrations of linoleic acid (LA) and alpha-linolenic acid (LNA) were high. The ratio of LA to LNA, (21.6), was higher than has been reported from other countries. The concentration of docosa-hexaenoic acid (DHA) was low and the ratio of arachidonic acid (AA) to DHA was much higher (2.8) than recommended and similar to that found in vegans. The concentrations of AA and DHA in the milk correlated positively with the infants' weight gain at the third month (p<0.05) and of DHA with length gain at the first and third months (p<0.01). Since the concentration of AA and, particularly, DHA in the milk declined during lactation, DHA deficiency may appear after 3-4 mo of age in breastfed Chinese rural infants. Further studies of Chinese rural mother-infant pairs are necessary to prove whether supplementation with suitable sources of AA and DHA, such as fish oil, should be recommended as lactation is lengthened to ensure optimal infant growth and development.

Age Factors↗

PubMed1999

Genetic analysis of nif regulatory genes by utilizing the yeast two-hybrid system detected formation of a NifL-NifA complex that is implicated in regulated expression of nif genes.

In diazotrophic organisms, nitrogenase synthesis and activity are tightly regulated. Two genes, nifL and nifA, are implicated as playing a major role in this regulation. NifA is a transcriptional activator, and its activity is inhibited by NifL in response to availability of excess fixed nitrogen and high O(2) tension. It was postulated that NifL binds to NifA to inhibit NifA-mediated transcriptional activation of nif genes. Mutational analysis combined with transcriptional activation studies clearly is in agreement with the proposal that NifL interacts with NifA. However, several attempts to identify NifA-NifL interactions by using methods such as coimmunoprecipitations and chemical cross-linking experiments failed to detect direct interactions between these proteins. Here we have taken a genetic approach, the use of a yeast two-hybrid protein-protein interaction assay system, to investigate NifL interaction with NifA. A DNA fragment corresponding to the kinase-like domain of nifL was PCR amplified and was used to generate translation fusions with the DNA binding domain and the DNA activation domain of the yeast transcriptional activator GAL4 in yeast two-hybrid vectors. Similarly, a DNA fragment corresponding to the catalytic domain of nifA was PCR amplified and used to generate translation fusions with the DNA-binding domain and the DNA-activation domain of GAL4 in yeast two-hybrid vectors. After introducing appropriate plasmid combinations in yeast cells, the existance of direct interaction between NifA and NifL was analyzed with the MATCHMAKER yeast two-hybrid system by testing for the expression of lacZ and his3 genes. These analyses showed that the kinase-like domain of NifL directly interacts with the catalytic domain of NifA.

Amino Acid Sequence↗

PubMed1999

[The effect of vasectomy on spermatogenesis and proliferation of rat spermatogenic cell].

In order to explore the effect of vasectomy on spermatogenesis and proliferation of rat spermatogenic cell, we detected the DNA content of rat spermatogenic cells by flow cytometry at 4 weeks and 8 weeks after vasectomy. The results showed that the percentages of 1C cell group (spermatid and sperm) in the two experimental groups were apparently lower than that of the two control groups respectively (P < 0.05), but the percentage of 2C cell group (spermatogonium and spermotocyte) in the two experiment groups were much higher than those of the two control groups respectively (P < 0.05); The proliferation index (PI) of experimental groups was significantly lower than that of control groups (P < 0.05) and most of the cells in the experimental groups were on G0/G phase. The percentage of S-phase cell in the 8 weeks experimental group was obviously lower than that of the 4 weeks experimental group. These findings suggest that vasectomy in rat may affect many stages in spermatogenesis and delay the proliferation of spermatogenic cell.

Animals↗

PubMed1999

[Effect of immunologic treatment on lymphocyte subsets in patients with myasthenia gravis].

OBJECTIVE: To study the abnormalities of cellular immunity of myasthenia gravis (MG) and the effects of glucocorticoid (GC) therapy and thymectomy on lymphocyte subsets in patients with myasthenia gravis. METHODS: Phenotypic analysis of peripheral blood lymphocyte in 39 patients with MG and 18 healthy controls were completed using flow cytometry and immunofluorescence double-staining methods; changes of lymphocyte subsets were studied in 15 of the MG patients after GC therapy and 7 of the MG patients after thymectomy. RESULTS: (1) The percentage of pan-T (CD(+)(5)), pan-B (CD(+)(19)), traditional B cell, CD(+)(4)T cell in patients with MG did not significantly differ from that in healthy controls. An increase in CD(+)(8) cell was observed in MG patients as compared with normal controls. Among the CD(+)(4) T lymphocyte subsets, there was significant decrease of the percentage of CD(+)(4) CD(45)RA(+) (suppressor-inducer) and increase of the percentage of CD(+)(4) CD(45)RA(-) (helper-inducer) subsets in MG patients than controls; the percentage of CD(+)(5) B cell was significantly increased in MG patients as compared with that in healthy controls. (2) After GC therapy, the percentages of pan-T (CD(+)(5)), pan-B (CD(+)(19)), traditional B cell, CD(+)(4) T cell, CD(+)(8) T cell, CD(+)(4) CD(45)RA(-), CD(+)(5) B cell decreased. (3) After thymectomy, the proportions of T, B lymphocyte subsets did not change significantly. CONCLUSION: Not only changes of T cell subsets (CD(+)(4) and CD(+)(8) T lymphocyte subsets) were shown, but also there were abnormal changes of functional CD(+)(4) T lymphocyte subsets and CD(+)(5) B lymphocyte which may play an important role in the pathogenesis of MG. There is comprehensive inhibiting action of GC on cellular immunity in MG patients, while there is no such effect with thymectomy on the lymphocyte subsets in MG patients.

Adolescent↗