Biomedical subjects

L Deng

Publications and source records attributed to L Deng.

At least 55 records · Page 3Linked to original sources

PubMed2001

[Further study on cyclin D1 expression and function in human nasopharyngeal carcinoma cell line].

OBJECTIVE: In the present study, we tried to further elucidate the expression and function of cyclin D1 in human nasopharyngeal carcinoma (NPC) cell line. METHODS: Western blot was used in the analysis of D-type cyclin expression profile. The distribution of this protein was determined in the double parameter flow cytometry (FCM). The function of cyclin D1 in nasopharyngeal carcinoma cell line was determined in the antisense-oligonucleotied-cyclin D1 and antibody mediated knock-out experiments. RESULTS: The immunoblot analysis of two NPC cell lines showed that the spectra of D-type in NPC cell lines founded their expressions in D1, D2, and D3. It also suggested that the overexpression of cyclin D1 could lead to deregulation of G1/S control. An additional double parameter FCM showed a characteristic variation in HNE1 which was consistent with the cell-cycle oscillation and the peak level expressed in G0/G1 phase and the lowest level in the S-phase. Two experimental approaches aimed at specific functional knock-out of cyclin D1 were employed; the cyclin D1-ASPODN experiment showed an inhibition of cyclin D1 expression at mRNA level and a down-regulated cyclin D1 partial inhibition of progression into S phase, resulting in cell growth arrest while the electroporation of neutralizing antibodies demonstrated a specific delay of S-phase entry in HNE1 and the requirement for the cyclin D1 in cell cycle progression of HNE1 cell line in regulation of G1/S. CONCLUSION: These data reveal that the cell-cycle regulatory function of cyclin D1 is essential for cell cycle progression in the G1 of nasopharyngeal carcinoma cell line and suggest that cyclin D1 play an important role in NPC carcinogenesis.

Cell Cycle↗

PubMed2001

[Detection of TTV DNA from high background nuclear acid samples using specific nuclear acid captured-polymerase chain reaction].

OBJECTIVE: To improve the amplification of non-specificity of TTV DNA detection using PCR from high background nuclear acids because of its low level in samples. METHODS: Specific nuclear acid captured PCR(SNAC-PCR) was established through capturing TTV DNA onto microplate with specific nuclear acid by means of avidin-biotin system, then amplifying captured TTV DNA as usual. Its specificity was evaluated by comparing with the detection of routine extraction samples using nested-PCR with or without subsequent DNA sequencing in 10 samples of sera and liver tissues in pairs. RESULTS: Positive results were obtained using routine protocol in 4 and 9 out of 10 samples of the sera and the liver tissues respectively. The PCR products were proved to be specific using RFLP analysis with Kpn I. However, positive results were found in only 2/10 and 3/10 of samples of sera and liver tissue using DNA sequencing. The results of SNAC-PCR were the same as those of routine protocol with subsequent DNA sequencing. Furthermore, sharper electrophoresis bands were obtained. CONCLUSION: SNAC-PCR could dramatically increase the specificity of the detection of TTV DNA, especially in high background nuclear acid samples. It can be widely used in the detection of other pathogens with low seral level or in high background nuclear acid samples. The level of TTV infection might be over-evaluated by currently used TTV DNA detection when the PCR products were not confirmed by DNA sequencing.

Base Sequence↗

PubMed2001

[Regulation of XW630 on TGF-beta 1 and collagen I gene expression in osteoblasts].

OBJECTIVE: The purpose of this part of study was to investigate the mechanism of a new anti-osteoporotic drug, XW630 on promoting osteogenic activity. METHODS: Total RNA were isolated from osteoblasts of the mice after the reaction with XW630 in 24 and 72 hours, that were hybridized with the extracted and digoxigenin-labeled TGF-beta 1 and the collagen I cDNA probes. RESULTS: XW630 significantly promoted TGF-beta 1 and collagen I gene expression in osteoblasts, and this drug was superior than other two kinds of estrogen in promoting osteogenic activity (P < 0.05). CONCLUSION: XW630 promote osteogenic activity of osteoblasts, probably by enhancing the mRNA expression of TGF-beta 1 and collagen I.

Animals↗

PubMed2001

[Growth and osteogenesis characteristics of cultured rat marrow stromal cells under bone induction condition].

OBJECTIVE: To investigate the growth and osteogenesis characteristics of cultured rat marrow stromal cells(rMSCs) under bone induction condition. METHODS: rMSCs were isolated from adult rat using density gradient separation method. The rMSCs attachment formed soon after seeding and grew into colonies with the appearance of fibroblastic cells. The osteogenic induction compound of Dex(10(-8) mol/L), beta-GP(10 mmol/L) and AA(50 micrograms/ml) was added to different passaged rMSCs and the proliferation and osteogenic differentiation of them was observed. RESULTS: The induction compound had strong effect on promoting proliferation, especially on that of further passaged rMSCs, and its stimulation effect on osteogenesis was also well proved with the expression of alkaline phosphatase (ALP) rising after one-week induction. Further subculturing from P1 to P6 led to the increase of proliferation activity of rMSCs. After three-week induction, mineral deposits appeared in the culture. CONCLUSION: The lower expression of ALP in P1 rMSCs and the increase of it in further passages or under induction condition suggest the rMSCs in our culture system are mainly undifferentiated osteoprogenitors.

Alkaline Phosphatase↗

PubMed2001

[Polycystic ovaraian syndrome and hyperinsulinemia].

OBJECTIVE: To identify the relationship between polycystic ovarian syndrome and hyperinsulinemia. METHODS: Age, body height, body weight, serum estradiol, progesterone, testosterone, follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), triiodothyronine (T3), tetraiodothyronine (T4), insulin, oral glucose tolerance test (OGTT), and cortisol were measured in 28 patients with polycystic ovarian syndrome (PCOS). RESULTS: Sxiteen patients (57.1% of total) had hyperinsulinemia. Among them one patient was found diabetic. No difference was noted in age, height, body weight, body mass index, serum estradiol, progesterone, testosterone, FSH, LH, PRL, T3, T4, cortisol, and glucose of OGTT between hyperinsulinemia and non-hyperinsulinemia patients. However the LH/FSH ratio (2.4 +/- 1.5) in hyperinsulinemia patients was significantly higher than 1.2 +/- 0.6 in non-hyperinsulinemia patients (P = 0.0125). In addition, the results showed that the fasting serum insulin, insulin after OGTT, the insulin/glucose ratio at 30, 120 minutes, 180 minutes, the area under curve of and the insulin, insulin resistant index (IRI) were higher in hyperinsulinemia pateints than in non-hyperinsulinemia patients respectively. Furthermore, the fasting insulin level, insulin/glucose ratio were found to be negatively related to insulin sensitivity index and positively related with beta-cell functioning index. CONCLUSION: Hyperinsulinemia and insulin resistance accompany with some of the PCOS patients in China. Further investigations will be necessary to clarify the relevant mechanisms.

Adult↗

PubMed2001

[Molecular spectral diagnosis of star forming regions].

Stars are the basic building blocks of our universe, therefore it is one of the most important research topics in astrophysics to understand the origin and the early evolution of these objects. The current picture is that stars are formed during the collapse of a large enough self-gravitating interstellar molecular cloud. The early collapse gives birth to a fetus of a star, which is surrounded by a rotating accretion disk. The proto-star accretes interstellar matter through the disk which in turn transfer the accumulated matter to the central proto-star, then the star gets weight during the process. Observation shows that gorgeous ejection of matter always come along with the accretion process. In the presence of disks, these outflows usually escape from the system along the axis of the disk, forming so called bipolar outflows. Typical tracers of these activities are rich molecules such as CO, SiC2, C3H, C3H2 etc. Observationally, such typical molecular outflows can be detected using Doppler effect by spectroscopic measurements. Using the 13.7 m radio telescope in Delingha station of Purple Mountain Observatory, we performed a survey for 12 low temperature IRAS objects, some of the sources show high velocity properties. Detailed analysis of the Doppler profiles of IRS34 is presented. Star forming activities are clearly seen in this field.

English Abstract↗

PubMed2000

Acetylation of HIV-1 Tat by CBP/P300 increases transcription of integrated HIV-1 genome and enhances binding to core histones.

The HIV-1 Tat protein is required for viral replication and is a potent stimulator of viral transcription. Although Tat has been extensively studied in various reductive paradigms, to date there is little information as to how this activator mediates transcription from natural nucleosomally packaged long terminal repeats. Here we show that CREB-binding protein (CBP)/p300 interacts with the HIV-1 Tat protein and serves as a coactivator of Tat-dependent HIV-1 gene expression on an integrated HIV-1 provirus. The site of acetylation of Tat was mapped to the double-lysine motif in a highly conserved region, (49)RKKRRQ(54), of the basic RNA-binding motif of Tat. Using HLM1 cells (HIV-1(+)/Tat(-)), which contain a single copy of full-length HIV-1 provirus with a triple termination codon at the first AUG of the Tat gene, we find that only wild type, and not K50A, K51A, or K50A/K51A alone or in combination of ectopic CBP/p300, is able to produce full-length infectious virions, as measured by p24 gag ELISAs. Tat binds CBP/p300 in the minimal histone acetyltransferase domain (1253-1710) and the binding is stable up to 0.85 M salt wash conditions. Interestingly, wild-type peptide 41-54, and not other Tat peptides, changes the conformation of the CBP/p300 such that it can acquire and bind better to basal factors such as TBP and TFIIB, indicating that Tat may influence the transcription machinery by helping CBP/p300 to recruit new partners into the transcription machinery. Finally, using biotinylated wild-type or acetylated peptides, we find that acetylation decreases Tat's ability to bind the TAR RNA element, as well as to bind basal factors such as TBP, CBP, Core-Pol II, or cyclin T. However, the acetylated Tat peptide is able to bind to core histones on a nucleosome assembled HIV-1 proviral DNA.

Acetylation↗

PubMed2000

Gene expression array of HTLV type 1-infected T cells: Up-regulation of transcription factors and cell cycle genes.

By utilizing a human cDNA expression array blot (588 genes), we have observed overexpression of various transcription factors, cell cycle regulated kinases, and DNA repair genes in HTLV-1-infected T cells. One of the genes of interest, and focus in this study, is the cyclin-dependent kinase inhibitor, p21/waf1. The p21/waf1 transcription and protein is overexpressed in all HTLV-1-infected cell lines tested as well as ATL and HAM/TSP patient samples. While p21/waf1 has been shown to display a selectivity for G(1)/S cyclin/cdk complexes, we have observed p21/waf1 to be complexed with cyclin A/cdk2. Functionally, the association of p21/cyclin A/cdk2 decreased the histone H1 phosphorylation in vitro, as observed in immunoprecipitations followed by kinase assays, as well as affecting other substrates such as the C-terminus of Rb protein involved in c-Abl and HDAC1 regulation. Wild-type, but not a mutant form (M47) of Tax, was found to be able to transactivate the p21/waf1 promoter in a p53-independent manner. We found that the minimal p21/waf1 promoter (-49 to +49 sequence) was activated by Tax and the minimal promoter contained two E2A transcription factor binding sites located between the TATA box and the initiation site. E2A proteins, E12 and E47, as well as a related helix-loop-helix protein, HEB, are all up-regulated in HTLV-1-infected T cells. When using band shift analysis, we found that only the E1 site (overlapping the transcription start site) was a functional DNA binding site. By using a chromatin immunoprecipitation (ChIP) assay, we observed that histone H4, and not histone H3, was acetylated from the endogenous p21/waf1 promoter in vivo, implying that CBP/p300, and not the SAGA complex, was critical in complexing with E2A in up-regulation of p21/waf1 in HTLV-1-infected cells.

Basic Helix-Loop-Helix Proteins↗

PubMed2000

[The effects of exogenous p16 expression on CDK4, Cyclin D1 and pRb in nasopharyngeal carcinoma cell lines].

OBJECTIVE: To investigate the effect of exogenous p16 on Cyclin D1, CDK4 and pRb, and to explore the mechanism of the growth suppression of p16 in nasopharyngeal carcinoma cell lines. METHODS: The curve of cell growth rate in three kinds of HNE1, # 3-2 and # 4-2 cell lines was analyzed and their double time was compared. Then the distribution of the cell cycle was detected by flow cytometry. The expression of p16 and the effect of exogenous p16 expression on CDK4, Cyclin D1 and pRb are studied by means of Western Blot. RESULTS: As compared with HNE1, # 3-2 and # 4-2 showed a longer double time(23.4 h vs 28.8 h, 31.2 h). # 4-2 showed a significant accumulation of cells in G0/G1 phase(P < 0.01) and decreasing in S phase(P < 0.05) while HNE1 and # 3-2 had no obvious difference(P > 0.05). Cyclin D1 expression was upregulated in # 4-2 but downregulated in # 3-2 by exogenous expressed p16. No obvious difference on CDK4 expression was found. Hypophosphorylated pRb was detected in three cell lines. The expression was stronger in # 4-2, and # 3-2 than that in HNE1 and Hela. Hyperphosphorylated pRb was also detected in HNE1. CONCLUSION: Exogenous p16 expression may arrest cell cycle in G0/G1 phase and suppress cell growth. The major mechanism is not to regulate the level of the expression of CDK4. There might be a threshold in p16 regulating Cyclin D1 expression. However, the final result contributes to the inhibition of pRb phosphorylation.

Cell Cycle↗

PubMed2000

L-750355, a human beta3-adrenoceptor agonist; in vitro pharmacology and profile of activity in vivo in the rhesus monkey.

The profile of in vitro and in vivo biology of a human beta3-adrenoceptor agonist, (S)-N-[4-[2-[[3[(2-amino-5-pyridinyl)oxy]-2-hydroxy-propyl]amino]-eth yl]-phenyl]-4-isopropylbenzenesulfonamide, L-750355, is described. Using cloned human and rhesus beta1-, beta2- and beta3-adrenoceptors, expressed in Chinese hamster ovary (CHO) cells, L-750355 was shown to be a potent, albeit partial, agonist for the human (EC(50)=10 nM; % maximal receptor activation=49%) and rhesus (EC(50)=28 nM; % maximal receptor activation=34%) beta3-adrenoceptors. Furthermore, L-750355 stimulates lipolysis in rhesus adipocytes in vitro. L-750355 is a weak partial agonist (EC(50)=3.2 microM; % maximal receptor activation=33% ) for the human beta1-adrenoceptor but exhibits no agonist activity for rhesus beta1- or beta2-adrenoceptors of either human or rhesus origin. Administration of L-750355 to anesthetized rhesus monkeys, as a series of rising dose intravenous infusions, evokes dose-dependent glycerolemia and tachycardia with no change in mean arterial blood pressure or plasma potassium. The dose-response curve for L-750355-induced glycerolemia lies to the left of that for tachycardia. Propranolol, at a dose (0.3 mg/kg, i.v. ) that attenuates isoproterenol-induced changes in heart rate and glycerolemia, abolished L-750355-induced tachycardia but had no effect on L-750355-induced glycerolemia.

Adrenergic beta-3 Receptor Agonists↗

PubMed2000

Discovery of a potent, orally bioavailable beta(3) adrenergic receptor agonist, (R)-N-[4-[2-[[2-hydroxy-2-(3-pyridinyl)ethyl]amino]ethyl]phenyl]-4-[4 -[4-(trifluoromethyl)phenyl]thiazol-2-yl]benzenesulfonamide.

As part of our investigation into the development of orally bioavailable beta(3) adrenergic receptor agonists, we have identified a series of pyridylethanolamine analogues possessing a substituted thiazole benzenesulfonamide pharmacophore that are potent human beta(3) agonists with excellent selectivity against other human beta receptor subtypes. Several of these compounds also exhibited an improved pharmacokinetic profile in dogs. For example, thiazole sulfonamide 2e (R = 4-F(3)C-C(6)H(4)) is a potent full beta(3) agonist (EC(50) = 3.6 nM, 94% activation) with >600-fold selectivity over the human beta(1) and beta(2) receptors, which also displays good oral bioavailability in several mammalian species, as well as an extended duration of action.

Administration, Oral↗

PubMed2000

Tetrahydroisoquinoline derivatives containing a benzenesulfonamide moiety as potent, selective human beta3 adrenergic receptor agonists.

Tetrahydroisoquinoline derivatives containing a 4-(hexylureido)benzenesulfonamide were examined as human beta3 adrenergic receptor (AR) agonists. Notably, 4,4-biphenyl derivative 9 was a 6 nM full agonist of the beta3 AR. Naphthyloxy compound 18 (beta3 EC50 = 78 nM) did not activate the beta1 and beta2 ARs at 10 microM, and showed >1000-fold selectivity over binding to the beta1 and beta2 ARs.

Adrenergic beta-3 Receptor Antagonists↗

PubMed2000

Activation of the IkappaB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain.

TRAF6 is a signal transducer in the NF-kappaB pathway that activates IkappaB kinase (IKK) in response to proinflammatory cytokines. We have purified a heterodimeric protein complex that links TRAF6 to IKK activation. Peptide mass fingerprinting analysis reveals that this complex is composed of the ubiquitin conjugating enzyme Ubc13 and the Ubc-like protein Uev1A. We find that TRAF6, a RING domain protein, functions together with Ubc13/Uev1A to catalyze the synthesis of unique polyubiquitin chains linked through lysine-63 (K63) of ubiquitin. Blockade of this polyubiquitin chain synthesis, but not inhibition of the proteasome, prevents the activation of IKK by TRAF6. These results unveil a new regulatory function for ubiquitin, in which IKK is activated through the assembly of K63-linked polyubiquitin chains.

Amino Acid Sequence↗

PubMed2000

Inhibition of Tat-mediated transactivation of HIV-1 LTR transcription by polyamide nucleic acid targeted to TAR hairpin element.

Tat, an essential human immunodeficiency virus type 1 protein interacts with the transactivation response element (TAR) and stimulates transcription from the viral long-terminal repeat (LTR). Blockage of Tat-TAR interaction halts viral transcription and hence replication. We have found that polyamide nucleic acid (PNA), targeted to the TAR sequences of viral RNA genome is able to prevent Tat-TAR interaction by efficient sequestration of the TAR. Anti-TAR PNA competes for TAR and prevents Tat-mediated stimulation of HIV-1 LTR transcription in vitro but has no influence on the basal level of transcription in the absence of Tat. Using a reporter gene construct pHIV LTR-CAT and pCMV-Tat in cell culture, we have further shown that anti-TAR PNA is able to block Tat-mediated transactivation of HIV-1 LTR transcription in vivo as judged by the extent of LTR driven CAT gene expression in the absence and presence of anti-TAR PNA. Supplementation of 100 nM of anti-TAR PNA into the culture medium further enhances the suppression of transactivation. Nonspecific scrambled PNA had no influence on Tat-TAR interaction and LTR-driven CAT gene expression in cell culture. These results suggest that PNA targeted to the TAR sequence of the viral genome may be a potential inhibitor of HIV-1 gene expression.

Antiviral Agents↗

PubMed2000

Human beta3-adrenergic receptor agonists containing 1,2,3-triazole-substituted benzenesulfonamides.

Compounds containing a 1,2,3-triazole-substituted benzenesulfonamide were prepared and found to be potent and selective human beta3-adrenergic receptor agonists. The most interesting compound, trifluoromethylbenzyl analogue 12e (beta3 EC50 = 3.1 nM with >1500-fold selectivity over binding to both beta1- and beta2 receptors), stimulates lipolysis in the rhesus monkey (ED50 = 0.36 mg/kg) and is 25% orally bioavailable in the dog.

Administration, Oral↗

PubMed2000

Neurogenesis in the vomeronasal epithelium of adult rats: evidence for different mechanisms for growth and neuronal turnover.

The pattern of cell migration during neuronal turnover in the vomeronasal sensory epithelium (VN-SE) is controversial. In mice, proliferating cells were detected at the edges and were described as migrating to the center of the VN-SE. In rats, in addition to proliferating cells at the margins of the epithelium, dividing cells are also present along the entire basal lamina of the VN-SE. In marsupials, dividing cells have also been observed in the margins and in the center of the VN-SE, the latter of which migrate vertically and become neurons. To investigate whether the process of neuronal turnover in placental mammals consists of horizontal and/or vertical migration, and whether or not this process is common to mammals, adult rats were injected with bromodeoxyuridine (BrdU) and allowed to survive for different periods of time. The distribution of BrdU-labeled cells in the horizontal and vertical dimension of the VN-SE was analyzed as a function of time. Both horizontal and vertical migrations of BrdU-labeled cells were detected. Since cells in the center of the VN-SE migrate vertically, and, as demonstrated by coexpression of markers of neuronal maturity and BrdU, become mature one day after undergoing mitosis, it is very likely that these cells participate in neuronal turnover. Conversely, because cells in the margins of the VN-SE stop migrating horizontally on day 14 before they have reached the center of the VN-SE, and since the VN-SE continues to grow during adulthood, it is likely that most of these latter cells constitute pools for growth.

Animals↗

PubMed2000

Potent, selective 3-pyridylethanolamine beta3 adrenergic receptor agonists possessing a thiazole benzenesulfonamide pharmacophore.

A series of thiazole benzenesulfonamide-substituted 3-pyridylethanolamines was prepared and evaluated for their human beta3 adrenergic receptor agonist activity. Incorporation of aryl and heteroaryl substitution in the 4-position of the thiazole ring resulted in a number of highly potent and selective beta3 agonists. Results of preliminary in vivo evaluation of several of these compounds is described.

Adrenergic beta-3 Receptor Agonists↗

PubMed2000

Imaging the phase of an evolving bose-einstein condensate wave function

We demonstrate a spatially resolved autocorrelation measurement with a Bose-Einstein condensate and measure the evolution of the spatial profile of its quantum mechanical phase. Upon release of the condensate from the magnetic trap, its phase develops a form that we measure to be quadratic in the spatial coordinate. Our experiments also reveal the effects of the repulsive interaction between two overlapping condensate wave packets and we measure the small momentum they impart to each other.

Journal Article↗