Biomedical subjects

Y Q Chen

Publications and source records attributed to Y Q Chen.

At least 37 records · Page 2Linked to original sources

PubMed2001

[Clinical study on effect of ginsenoside in inducing rectal cancer cell apoptosis].

OBJECTIVE: To explore the effect of ginsenoside in inducing rectal cancer cell apoptosis. METHODS: Fifty patients of rectal cancer diagnosed by pathological examination were randomly divided into two groups, 35 in the treated group and 15 in the control group. The treated group received retention enema with 84.5% ginsenoside for 4-6 hrs every day, 6-8 days consecutively before surgical operation. The control group also received retention enema in the same way but with normal saline instead of ginsenoside. Fresh sample was taken during the operation and examined by electron microscopy. RESULTS: Symptoms, such as frequent defecation, hematochezia and tenesmus, were palliated in most patients (25/35) and abdominal pain relieved in all the 7 cases of incomplete intestinal obstruction in the treated group. Electron microscopic examination showed that cell apoptosis was observed in cancer sample of 23 cases among the 35 cases of the treated group, amounting to 65.7%. While in the control group, above-mentioned changes were not observed at all. CONCLUSION: Ginsenoside has the effect of inducing apoptosis in rectal cancer patients.

Administration, Rectal↗

PubMed2001

[Synthesis of benzodihydropyran derivatives and evaluation of their preliminary biological activities on bone and vascular tissues].

AIM: To screen optimal drugs against postmenopausal osteoporosis with cardiovascular protective activities. METHODS: A series of benzodihydropyran derivatives were designed and synthesized in view of comprehensive observations of raloxifene and ipriflavone. The antiosteoporosis activities of compounds a-e (10(-7) mol.L-1) on the proliferation of human osteoblast cell HOS TE85 were studied. The cardiovascular protective activities were evaluated by observing their effects on proliferation of human vascular endothelium cell ECV-304 and their protective effects on ECV-304 damaged by H2O2. RESULTS: Their structures were determined by spectrums. Compounds a, b and c (10(-7) mol.L-1) were shown to significantly help proliferation of HOS TE85. In addition, b, d and e (10(-8) mol.L-1) helped proliferation of ECV-304 significantly. Compounds b and c (10(-6) mol.L-1) showed strong protective activity on ECV-304 damaged by H2O2. Compounds b and c shifted the KCl dose-response curves to the right and decreased the maximal response. CONCLUSION: Compounds b and c showed some bone and vascular protective activities which benefit postmenopausal and cardiovascular diseases.

Animals↗

PubMed2001

[Selection of high taxol content cell lines of Taxus yunnanensis Cheng et L. K. Fu].

OBJECTIVE: To select high taxol content cell lines of Taxus yunnanensis. METHOD: Choosing different cell aggregates according to their color, texture, growth rate and secretion of colorful substances, culturing them separately, and further analyzing their growth rates and taxolcontents. RESULT AND CONCLUSION: Cell lines with higher taxol contents could be obtained by careful selection; those with darker color, lower growth rate and higher ratio of dry cell weight vs. fresh cell weight usually has higher taxol contents.

Cell Line↗

PubMed2000

Overexpression and activation of the RON receptor tyrosine kinase in a panel of human colorectal carcinoma cell lines.

RON is a receptor tyrosine kinase belonging to the MET proto-oncogene family. The purposes of this study are to determine the expression and activation of RON in a panel of human colon carcinoma cell lines. Western blotting showed that RON is barely detectable in normal and SV-40-transformed colon epithelial cells, but highly expressed and constitutively activated in several colon carcinoma cell lines including Colo201, HT-29, HCT116, and SW837. Moreover, a novel RON variant with a molecular mass of 160 kDa (RONDelta160) was identified from HT-29 cells. The cDNA encoding RONDelta160 has an in-frame deletion of 109 amino acids in the extracellular domain of the RON beta chain, which is caused by splicing out of two exons in the RON mRNA. No mutations were found in the kinase domain of the RON gene in five carcinoma cell lines screened. By expressing RON in colon epithelial cells, we found that RON activation increases cell motile-invasive activities and protects cells against apoptotic death. These data suggest that RON expression and activation are deregulated in colon carcinoma cell lines. By abnormal activation of RON, this receptor and its variant may regulate motile-invasive phenotypes of certain colon carcinoma cells in vivo.

Amino Acid Sequence↗

PubMed2000

NF-kappaB p65 (RelA) homodimer uses distinct mechanisms to recognize DNA targets.

BACKGROUND: The NF-kappaB family of dimeric transcription factors regulates the expression of several genes by binding to a variety of related DNA sequences. One of these dimers, p65(RelA), regulates a subclass of these targets. We have shown previously that p65 binds to the 5'-GGAA T TTTC-3' sequence asymmetrically. In that complex one subunit base specifically interacts with the preferred 5' half site and the other subunit binds non-specifically to the 3' half site. RESULTS: Here we describe the crystal structures of two new p65-DNA complexes. One complex contains a pseudosymmetric 5'-GGAA T TTCC-3' DNA sequence taken from the enhancer of the gene encoding interleukin 8 (IL-8) and the other contains the asymmetric 5'-GGAA T TCCC-3' target DNA taken from the enhancer of the gene encoding type VII collagen. As expected, the global positioning of the dimer on both DNA targets is roughly symmetric, however, the hydrogen-bonding patterns at the protein-DNA interfaces differ significantly. One of the p65 monomers in complex with the asymmetric DNA binds to an extra base pair located immediately upstream of the 5'-GGAA-3' half site. We also show that p65 binds to these targets with almost equal affinity and that different residues have variable roles in binding different kappaB targets. CONCLUSIONS: Taken together, these structures reveal that p65 exhibits the unique capability to specifically bind DNA targets of variable lengths from four to ten base pairs. Also, the small protein segment Arg41-Ser42-Ala43 is at least partially responsible for flexibility in DNA-binding modes.

Amino Acid Sequence↗

PubMed2000

Regulation of the RON receptor tyrosine kinase expression in macrophages: blocking the RON gene transcription by endotoxin-induced nitric oxide.

Previous studies have shown that activation of the RON receptor tyrosine kinase inhibits inducible NO production in murine peritoneal macrophages. The purpose of this study is to determine whether inflammatory mediators such as LPS, IFN-gamma, and TNF-alpha regulate RON expression. Western blot analysis showed that RON expression is reduced in peritoneal macrophages collected from mice injected with a low dose of LPS. The inhibition was seen as early as 8 h after LPS challenge. Experiments in vitro also demonstrated that the levels of the RON mRNA and protein are diminished in cultured peritoneal macrophages following LPS stimulation. TNF-alpha plus IFN-gamma abrogated macrophage RON expression, although individual cytokines had no significant effect. Because LPS and TNF-alpha plus IFN-gamma induce NO production, we reasoned that NO might be involved in the RON inhibition. Two NO donors, S-nitroglutathione (GSNO) and (+/-)-S-nitroso-N-acetylpenicillamine (SNAP), directly inhibited macrophage RON expression when added to the cell cultures. Blocking NO production by NO inhibitors like TGF-beta prevented the LPS-mediated inhibitory effect. In Raw264.7 cells transiently transfected with a report vector, GSNO or SNAP inhibited the luciferase activities driven by the RON gene promoter. Moreover, GSNO or SNAP inhibited the macrophage-stimulating protein-induced RON phosphorylation and macrophage migration. We concluded from these data that RON expression in macrophages is regulated during inflammation. LPS and TNF-alpha plus IFN-gamma are capable of down-regulating RON expression through induction of NO production. The inhibitory effect of NO is mediated by suppression of the RON gene promoter activities.

Animals↗

PubMed2000

Requirement of both tyrosine residues 1330 and 1337 in the C-terminal tail of the RON receptor tyrosine kinase for epithelial cell scattering and migration.

RON is a receptor tyrosine kinase that mediates cell scattering, migration, and tubular formation. This study focused on the function of two tyrosines, Y1330 and Y1337, in the C-terminus of RON in regulating epithelial cell scattering and migration. Substitution of both tyrosine residues with phenylalanine causes complete loss of cell scattering and migration in kidney 293 cells. In contrast, single mutation of either tyrosine residue has no effect. We found that mutation at Y1330 or Y1337 alone does not significantly affect the association of RON with PI-3 kinase, whereas a double mutation abolishes the recruitment of substrates. RON-mediated cell migration was inhibited by PI-3 kinase inhibitor wortmannin. This effect was also achieved by a dominant inhibitory p85 of PI-3 kinase. We conclude that Y1330 and Y1337 are required for RON-mediated cell motility. By associating with PI-3 kinase, the Y1330-Y1337 docking site plays a critical role in transducing motile signals of RON.

Amino Acid Substitution↗

PubMed2000

H-ras, but not N-ras, induces an invasive phenotype in human breast epithelial cells: a role for MMP-2 in the H-ras-induced invasive phenotype.

Elevated p21ras expression is associated with tumor aggressiveness in breast cancer including the extent of invasion into fat tissues, infiltration into lymphatic vessels and tumor recurrence. In the present study, we have examined the roles of H-ras and N-ras, members of the human ras gene family, in the pathogenesis of breast cancer. We show that H-ras, but not N-ras, induces an invasive phenotype in human breast epithelial cells (MCF10A) as determined by the Matrigel invasion assay, whereas both H-ras and N-ras induce anchorage-independent growth, as shown by soft agar assay. We examined the effects of H-ras and N-ras activation on the expression of MMP-2 and MMP-9, which can degrade type IV collagen, the major structural collagen of the basement membrane. We show that MMP-2 is efficiently induced by H-ras, whereas MMP-9 induction is more prominent in N-ras-activated MCF10A cells. We also show that H-ras-mediated invasiveness is significantly inhibited when the expression of MMP-2 is down-regulated, using an oligodeoxyribonucleotide complementary to the MMP-2 mRNA, or when MMP-2 activity is blocked by its inhibitor TIMP-2 (tissue inhibitors of matrix metalloproteinase-2). Our results show that the H-ras-induced invasive phenotype is associated more closely with the expression of MMP-2 in human breast epithelial cells, rather than the induction of MMP-9 expression, as shown previously for rat embryonic fibroblasts.

Breast Neoplasms↗

PubMed2000

Estimating a treatment effect with the accelerated hazards models.

In randomized clinical trials, when the outcome of interest is time to event, the proportional hazards model or the accelerated failure time model is often used to identify a treatment effect. In this article, we discuss a simple alternative called the accelerated hazards model in which the treatment effect is characterized as the hazard progression time ratio, when the treatment is believed to accelerate or decelerate the underlying hazard progression through time. Survival data from an actual randomized placebo-controlled trial, which evaluates the effectiveness of biodegradable polymers with carmustine to treat malignant gliomas, is used for illustration.

Antineoplastic Agents, Alkylating↗

PubMed2000

Nonparametric and semiparametric trend analysis for stratified recurrence times.

Recurrent event data are frequently encountered in longitudinal follow-up studies when the occurrences of multiple events are considered as the major outcomes. Suppose that the recurrent events are of the same type and the variable of interest is the recurrence time between successive events. In many applications, the distributional pattern of recurrence times can be used as an index for the progression of a disease. Such a distributional pattern is important for understanding the natural history of a disease or for confirming long-term treatment effect. In this article, we discuss and define the comparability of recurrence times. Nonparametric and semiparametric methods are developed for testing trend of recurrence time distributions and estimating trend parameters in regression models. The construction of the methods is based on comparable recurrence times from stratified data. A real data example is presented to illustrate the use of methodology.

Biometry↗

PubMed2000

Tissue hypoxia inhibits prostaglandin and nitric oxide production and prevents ductus arteriosus reopening.

Regulation of ductus arteriosus (DA) tension depends on a balance between oxygen-induced constriction and PG and nitric oxide (NO)-mediated relaxation. After birth, increasing Pa(O(2)) produces DA constriction. However, as the full-term ductus constricts, it develops severe tissue hypoxia in its inner vessel wall (oxygen concentration <0.2%). We used isolated rings of fetal lamb DA to determine why the constricted ductus does not relax and reopen as it becomes hypoxic. We used a modification of the 2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl) acetamide (EF5) technique (Clyman RI, Chan CY, Mauray F, Chen YQ, Cox W, Seidner SR, Lord EM, Weiss H, Wale N, Evan SM, and Koch CJ. Pediatr Res 45: 19-29, 1999) to determine mean tissue oxygen concentration. A decrease in the ductus' mean tissue oxygen concentration from 1.4 to 0.1% lowers the isometric tone of the ductus by 15 +/- 10% of its maximal active tension (the maximal tension that can be produced by the ductus). Although decreases in oxygen concentration diminish ductus tension, most of the vasoconstrictor tone in the ductus is independent of ambient oxygen concentration. This oxygen-independent tone is equivalent to 64 +/- 10% of the maximal active tension. At mean tissue oxygen concentrations >0.2%, endogenous PGs and NO inhibit more than 40% of the active tension developed by the ductus. However, when tissue oxygen concentrations drop below 0.2%, the constitutive relaxation of the ductus by endogenous PGs and NO is lost. In the absence of PG and NO production, tension increases to a level normally observed only after treatment of the ductus with indomethacin and nitro-L-arginine methyl ester (inhibitors of PG and NO production). Therefore, under conditions of severe hypoxia (tissue oxygen concentration <0.2% oxygen), the loss of PG- and NO-mediated relaxation more than compensates for the loss of oxygen-induced tension. We hypothesize that this increased ductus tone enables the vessel to remain closed as it undergoes tissue remodeling.

Animals↗

PubMed2000

Synergistic effect of adrenal steroids and angiotensin II on plasminogen activator inhibitor-1 production.

Recent data suggest an interaction between the renin-angiotensin-aldosterone system and fibrinolysis. Although previous work has focused on the effect of angiotensin II (Ang II) on plasminogen activator inhibitor (PAI-1) expression, the present study tests the hypothesis that aldosterone contributes to the regulation of PAI-1 expression. To test this hypothesis in vitro, luciferase reporter constructs containing the human PAI-1 promoter were transfected into rat aortic smooth muscle cells. Exposure of the cells to 100 nmol/L Ang II resulted in a 3-fold increase in luciferase activity. Neither 1 micromol/L dexamethasone nor 1 micromol/L aldosterone alone increased PAI-1 expression. However, both dexamethasone and aldosterone enhanced the effect of Ang II in a dose-dependent manner. This effect was abolished by mutation in the region of a putative glucocorticoid-responsive element. A similar interactive effect of Ang II and aldosterone was observed in cultured human umbilical vein endothelial cells. The time course of the effect of aldosterone on Ang II-induced PAI-1 expression was consistent with a classical mineralocorticoid receptor mechanism, and the effect of aldosterone on PAI-1 synthesis was attenuated by spironolactone. To determine whether aldosterone affected PAI-1 expression in vivo, we measured local venous PAI-1 antigen concentrations in six patients with primary hyperaldosteronism undergoing selective adrenal vein sampling. PAI-1 antigen, but not tissue plasminogen activator antigen, concentrations were significantly higher in adrenal venous blood than in peripheral venous blood. Taken together, these data support the hypothesis that aldosterone modulates the effect of Ang II on PAI-1 expression in vitro and in vivo in humans.

Adrenal Cortex Hormones↗

PubMed2000

[Effects of culture conditions on callus growth and taxol formation of Taxus yunnanensis Cheng et L.K.Fu].

OBJECTIVE: To seek culture conditions suitable for the callus growth and taxol formation of Taxus yunnanensis. METHOD: Determining the growth index and taxol content in the callus cultured under different illumination conditions or on different media. RESULTS AND CONCLUSION: Darkness, the combination of 0.1 mg.L-1BAP and 2,4-D 1.0 mg.L-1, and B5, DCR or 6,7-V basal medium were favorable for the callus growth and taxol formation of Taxus yunnanensis; high NO3- concentration in the medium promoted callus growth, and high NH4+ concentration greatly inhibited callus growth, but dramatically increased taxol content.

Culture Media↗

PubMed2000

[Determination of chiral purity of synthetic amino acids by high performance liquid chromatography].

A series of synthetic beta-heterocyclic and beta-naphthyl alanine and aromatic substituted phenylalanine were separated by reversed-phase HPLC as diastereomeric derivatives obtained after derivatization with (1-fluoro-2,4-dinitrophenyl)-5-L-valinamide (FDNP-Val-NH2). The separation was performed with gradient elution. The mobile phase A was acetonitrile containing 0.1% trifluoroacetic acid and mobile phase B was 0.1% trifluoroacetic acid. According to the hydrophobicity of the side chain of the amino acids, 6 different linear gradient programs were selected and all gradients completed in 20 minutes. Good resolution was obtained for neutral amino acids. All L-L diastereomer of neutral amino acids (the first letter refers to the configuration of amino acid) were eluted faster than the D-L diastereomer with retention times less than 16 minutes. In the same gradient eluting system, the hydrophobic parameter (pi value) and the position of the substituent in benzene ring of phenylalanine also influence the retention time of diastereomers. Usually the retention time of the phenylalanine with a large pi value was longer than that with a small one. Among the three phenylalanines containing the same substituent, compounds with the substituent in 2-postion gave the shortest retention time. In the case of basic amino acids, the D-L diastereomers eluted prior to the L-L diastereomers and showed rather poor resolution. The chiral purity (ee) of the optical active amino acids (synthetic DL-amino acids after resolution by different enzymes) were calculated with the equation as follows: ee = [[A(L- or D-) - A(D- or L-)]/[A(L-) + A(D-)]] x 100% (L- or D-), A: peak area; L-, D-: amino acid diastereomer.

Amino Acids↗

PubMed1999

Induction of apoptosis and G2/M cell cycle arrest by DCC.

The Deleted in Colorectal Cancer gene (DCC) encodes a cell surface receptor that belongs to the Ig superfamily. Inactivation of the DCC gene has been implicated in human tumor progression. However, little is known about the biological function of the DCC protein. In the present study, we demonstrated that expression of DCC activated caspase-3 and programmed cell death, or induced G2/M cell cycle arrest in tumor cells. In some cell lines, apoptosis was evident within 24 h of DCC expression. Timing of the appearance of apoptotic cells coincided with that of the cleavage of poly (ADP-ribose) polymerase, a substrate of caspase-3. Expression of the apoptosis inhibitory gene Bcl-2 was not able to abrogate the DCC-induced apoptosis. In the G2/M cycle arrest cells, cdk1 activity was inhibited. Our results suggest that the DCC protein may transduce signals resulting in activation of caspases or inhibition of Cdk1. These data provide a possible mechanism by which DCC suppresses tumorigenesis.

Apoptosis↗

PubMed1999

Inhaled IL-5 increases concentrations of soluble intracellular adhesion molecule-1 in sputum from atopic asthmatic subjects.

BACKGROUND: Both IL-5 and intracellular adhesion molecule-1 (ICAM-1) have been shown to play important roles in the production of allergic inflammation, including atopic asthma. OBJECTIVE: The aim of this study was to investigate the effects of recombinant human IL-5 on changes of soluble (s) ICAM-1 concentrations in induced sputum from allergic asthmatic subjects. METHODS: In a randomized, double-blind, placebo-controlled study design, 8 nonsmoking patients with allergic asthma and 6 nonallergic normal subjects were administered recombinant human IL-5 by nebulization, and the concentrations of sICAM-1 in induced sputum from each subject were determined before and at 2, 24, 48, and 72 hours after inhalation. RESULTS: sICAM-1 levels in sputum within the control group did not appear to change from baseline at any time throughout the study. In allergic asthmatic subjects vehicle challenge was not able to cause any changes in sputum sICAM-1 concentrations. However, there were increases in sputum sICAM-1 concentrations after IL-5 inhalation, which increased with time to significantly greater levels than those at baseline, reaching a maximum at 48 hours and lasting no less than 72 hours. The concentrations of sICAM-1 in sputum after IL-5 challenge exceeded levels that could be accounted for by passive transudation from the circulation on the basis of the magnitude of increases in sputum albumin concentration. CONCLUSION: Our results indicated that IL-5 inhalation was capable of inducing an elevation of sputum sICAM-1 concentration by stimulating its local release in allergic asthmatic, but not in nonallergic normal, subjects.

Administration, Inhalation↗

PubMed1999

Cyclooxygenase-2 plays a significant role in regulating the tone of the fetal lamb ductus arteriosus.

Nonselective cyclooxygenase (COX) inhibitors are potent tocolytic agents but have adverse effects on the fetal ductus arteriosus. We hypothesized that COX-2 inhibitors may not affect the ductus if the predominant COX isoform is COX-1. To examine this hypothesis, we used ductus arteriosus obtained from late-gestation fetal lambs. In contrast to our hypothesis, fetal lamb ductus arteriosus expressed both COX-1- and COX-2-immunoreactive protein (by Western analysis). Although COX-1 was found in both endothelial and smooth muscle cells, COX-2 was found only in the endothelial cells lining the ductus lumen (by immunohistochemistry). The relative contribution of COX-1 and COX-2 to PGE2 synthesis was consistent with the immunohistochemical results: in the intact ductus, PGE2 formation was catalyzed by both COX-1 and COX-2 in equivalent proportions; in the endothelium-denuded ductus, COX-2 no longer played a significant role in PGE2 synthesis. NS-398, a selective inhibitor of COX-2, was 66% as effective as the selective COX-1 inhibitor valeryl salicylate and the nonselective COX inhibitor indomethacin in causing contraction of the ductus in vitro. At this time, caution should be used when recommending COX-2 inhibitors for use in pregnant women.

Animals↗

PubMed1999

Permanent anatomic closure of the ductus arteriosus in newborn baboons: the roles of postnatal constriction, hypoxia, and gestation.

Permanent closure of the ductus arteriosus requires loss of cells from the muscle media and development of neointimal mounds, composed in part of proliferating endothelial cells. We hypothesized that postnatal ductus constriction produces hypoxia of the inner vessel wall; we also hypothesized that hypoxia might lead to cell death and the production of vascular endothelial cell growth factor (VEGF), a hypoxia-inducible growth factor that stimulates endothelial proliferation. We mapped the distribution of hypoxia in newborn baboons and correlated it with the appearance of cell death (TUNEL technique), VEGF expression, and endothelial proliferation (proliferating cell nuclear antigen expression). In the full-term baboon (n=10), the ductus was functionally closed on Doppler examination by 24 h after delivery. Regions of the ductus where the lumen was most constricted were associated with moderate/intense hypoxia; VEGF expression was increased in the hypoxic muscle media, and luminal endothelial cells, adjacent to the hypoxic media, were proliferating. Cells in the most hypoxic regions of the ductus wall were undergoing DNA fragmentation. In contrast, regions of the ductus with mild degrees of hypoxia had no evidence of cell death, VEGF expression, or endothelial proliferation. Cell death and endothelial proliferation seemed to be limited to regions of the full-term ductus experiencing moderate/intense hypoxia. In the premature baboon (67% gestation) (n=24), only 29% closed their ductus by Doppler examination before d 6. None of the premature baboons, including those with a closed ductus by Doppler, had evidence of moderate/intense hypoxia; also, there was no evidence of cell death, VEGF expression, endothelial proliferation, or neointima formation by d 6. Therefore, the premature ductus is resistant to developing hypoxia, even when its lumen is constricted; this may make it susceptible to later reopening.

Animals↗