Biomedical subjects

P Li

Publications and source records attributed to P Li.

At least 217 records · Page 12Linked to original sources

PubMed2000

[Application of RAPD in the taxonomy of the genus Fritillaria].

OBJECTIVE: To provide the molecular data for the right application of the Chinese medicine "Beimu". METHOD: Using the technology of RAPD, we studied the relationship among 12 samples of Beimu. RESULT: The total genomic DNA of all the samples are about 21 Kb in size. Among 20 primers used, the five primers can reatedly generate a certain specified amplified band type, 27 bands were recored from all amplified products and 25 polymorphic fragments were found in it. The size of amplified fragments is between 450 bp and 1904 bp. CONCLUSION: The similarity within species is higher than those between species. The relationship of Fritillaria anhuiensis and F. puqiensis is the farest, while F. thunbergii and F. puqiensis is the closest.

DNA Primers↗

PubMed2000

Experimental study of homium: YAG laser transmyocardial revascularization in acute ischemic settings with myocardial contrast echocardiography.

OBJECTIVE: To study the mechanism and effects of blood perfusion to the acute ischemic region of myocardium through Ho-YAG laser channels with myocardial contrast echocardiography. METHODS: To produce the model of acute myocardial ischemia, we partially ligated the left anterior decending (LAD) coronary artery of canine hearts between 1st. and 2nd. diagonal branches and then performed transmyocardial revascularization in this region with Ho - YAG laser. Myocardial contrast echocardiography was made with a new generation of ultrasound contrast agent and second harmonic imaging of this region before, after ischemia and after laser revascularization. Pictures were taken with "R" wave trigger skill. RESULTS: Acoustic density determing in the ischemia region (anterior wall) with MCE (myocardial contrast echocardiography) was obviously decreased (5.40 +/- 1.81) after the LAD was ligated,as compared with before (11.69 +/- 1.61, P < 0.01). It was increased remarkably after transmyocardial laser revascularization (TMLR) (11.2 +/- 2.01, P < 0.01) as compared with that when ischemia and approximated to that before ischemia (P > 0.05). There were no differences in acoustic density in the lateral wall (as control) among these comprehensive three periods (P > 0.05). Contrast in the laser region developed one cardiac cycle ahead of that in the non-ischemic normal region. CONCLUSION: Acute ischemic myocardium can be perfused by oxygenated blood from the left ventricle through Ho - YAG laser channels. Evidence of blood perfusion through laser channels during systolic phase was detected, and myocardial contrast ultrasonography using intravenous perfluorocarbon-exposed sonicated dextrose albumin may be regarded as a reliable method in the study of transmyocardial revascularization.

Animals↗

PubMed1999

Solution structure of the CIDE-N domain of CIDE-B and a model for CIDE-N/CIDE-N interactions in the DNA fragmentation pathway of apoptosis.

Apoptotic DNA fragmentation and chromatin condensation are mediated by the caspase-activated DFF40/ CAD nuclease, which is chaperoned and inhibited by DFF45/ICAD. CIDE proteins share a homologous regulatory CIDE-N domain with DFF40/CAD and DFF45/ ICAD. Here we report the solution structure of CIDE-N of human CIDE-B. We show that the CIDE-N of CIDE-B interacts with CIDE-N domains of both DFF40 and DFF45. The binding epitopes are similar and map to a highly charged bipolar surface region of CIDE-B. Furthermore, we demonstrate that the CIDE-N of CIDE-B regulates enzymatic activity of the DFF40/ DFF45 complex in vitro. Based on these results and mutagenesis data, we propose a model for the CIDE-N/ CIDE-N complex and discuss the role of this novel bipolar interaction in mediating downstream events of apoptosis.

Amino Acid Sequence↗

PubMed1999

Rapid and efficient cell-to-cell transmission of human immunodeficiency virus infection from monocyte-derived macrophages to peripheral blood lymphocytes.

Macrophages are considered of central importance in cell-to-cell transmission of human immunodeficiency virus (HIV) infection in vivo. In this report, we describe a novel cell-to-cell transmission model using HIV-infected monocyte-derived macrophages (MDMs) as donor cells and peripheral blood lymphocytes (PBLs) as recipients. Virus was transmitted during a 2-h coincubation period from intracellular or tightly cell-associated viral stores in adherent infected MDMs to nonadherent CD3(+) PBLs. Transmission required cell contact, but syncytia formation was not observed. HIV cell-to-cell transmission occurred in both allogeneic and autologous systems, and replication was higher in phytohemagglutinin (PHA)-stimulated than unstimulated recipient PBLs. In contrast, transmission of infection by cell-free virus was barely detectable without PHA stimulation of recipients, suggesting the cell-cell interaction may have provided stimuli to recipient cells in the cell-to-cell system. Viral DNA levels increased 5-24 h postmixing, and this increase was inhibited by pretreatment of cells with the reverse transcription inhibitor azidothymidine, indicating de novo reverse transcription was involved. Cell-to-cell transmission was more efficient than infection with cell-free virus released from donor MDMs, or 0.1 TCID(50)/cell cell-free viral challenge. This model provides a system to further investigate the mechanisms and characteristics of HIV cell-to-cell transmission between relevant primary cells that may be analogous to this important mode of virus spread in vivo.

Cells, Cultured↗

PubMed1999

A G protein gamma subunit-specific peptide inhibits muscarinic receptor signaling.

Muscarinic acetylcholine receptors modulate the function of a variety of effectors through heterotrimeric G proteins. A prenylated peptide specific to the G protein gamma5 subunit type inhibits G protein activation by the M2 muscarinic receptor in a reconstitution assay. Scrambling the amino acid sequence of the peptide significantly reduces the efficacy of the peptide. The peptide does not disrupt the G protein heterotrimer. In cultured sympathetic neurons, the gamma5 peptide inhibits modulation of Ca(2+) current by the M4 receptor. Peptide activity is specific, the scrambled peptide and peptides specific to two other members of the G protein gamma subunit family are significantly less effective. The gamma5 peptide has no effect on Ca(2+) current modulation by the alpha2-adrenergic and somatostatin receptors. In addition, the gamma5 peptide inhibits muscarinic receptor signaling in spinal cord slices with specificity. These results support a specific role for G protein gamma subunit types in signal transduction, most likely at the receptor-G protein interface.

Amino Acid Sequence↗

PubMed1999

Loss of synaptic depression in mammalian anterior cingulate cortex after amputation.

Two forms of activity-dependent long-term depression (LTD) in the CNS, as defined by their sensitivity to the blockade of NMDA receptors, are thought to be important in learning, memory, and development. Here, we report that NMDA receptor-independent LTD is the major form of long-term plasticity in the anterior cingulate cortex (ACC). Both L-type voltage-gated calcium channels and metabotropic glutamate receptors are required for inducing LTD. Amputation of a third hindpaw digit in an adult rat induced rapid expression of immediate early genes in the ACC bilaterally and caused a loss of LTD that persisted for at least 2 weeks. Our results suggest that synaptic LTD in the ACC may contribute to enhanced neuronal responses to subsequent somatosensory stimuli after amputation.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1999

Pre-column derivatization and gas chromatographic determination of alkaloids in bulbs of Fritillaria.

A method of precolumn derivatization GC with FID detection was developed for a simultaneous analysis of five major steroidal alkaloids of Fritillaria species, namely ebeiedine, ebeiedinone, verticine, verticinone and imperialine. Derivatization was carried out by trimethylsilylation of the hydroxyl-containing Fritillaria alkaloids to the corresponding trimethylsilylates with trimethylsilylimidazole. Reaction conditions were optimised and the alkaloids derivatives were characterised by on-line GC-MS. The validated GC method demonstrated a good linearity at the sampling ranges used. This analytical method is simple, convenient and reproducible. The developed assay was successfully applied to the determination of the major pharmacologically active alkaloids in three commonly used antitussive Fritillaria species: F. cirrhosa, F. thunbergii and F. pallidiflora.

Alkaloids↗

PubMed1999

Study of DFF45 in its role of chaperone and inhibitor: two independent inhibitory domains of DFF40 nuclease activity.

CAD/DFF40, the nuclease responsible for DNA fragmentation during apoptosis, exists as a heterodimeric complex with DFF45/ICAD. This study determines the molecular mechanisms of regulation of DFF40 via the chaperone and inhibition activities of DFF45. We analyze proteins corresponding to the fragments (D1, D2, and D3) of DFF45 generated by cleavage at the caspase consensus sites in DFF45. Either D1 or D2, as an isolated domain, is capable of inhibiting DFF40 nuclease activity while double domain fragments D1-2 and D2-3, as well as full-length DFF45, bind to DFF40 with high affinity and are much more effective inhibitors. The chaperone activity of DFF45 resides in part in its ability to maintain DFF40 as a soluble protein. In addition, D1 of DFF45 was found to be critical for the expression of active DFF40 in vivo, suggesting a role for DFF45 in binding nascent DFF40.

Apoptosis Regulatory Proteins↗

PubMed1999

Multiple domains of DFF45 bind synergistically to DFF40: roles of caspase cleavage and sequestration of activator domain of DFF40.

CAD/DFF40, the nuclease responsible for DNA fragmentation during apoptosis, exists as a heterodimeric complex with DFF45/ICAD. The study presented here augments the accompanying inhibition and chaperone study with an analysis of specific binding strengths and locations of DFF45 binding sites within DFF40. This allows us to show that DFF40/45 interaction is mediated by binding of three functional domains (D1, D2, and D3) of DFF45 to two domains (activator and catalytic) of DFF40. D1 binds exclusively to the activator domain and D2 binds to the catalytic domain of DFF40. Inhibition of DFF40 nuclease activity arises independently from D1 functional sequestration of the activator domain and D2 blockage of the catalytic domain of DFF40. The mechanism of caspase activation of DFF40 is the disruption of the synergistic binding activity of DFF45 domains to DFF40 after caspase recognition and cleavage of DFF45 in the context of a DFF45/40 complex.

Apoptosis Regulatory Proteins↗

PubMed1999

Nardosinone, a novel enhancer of nerve growth factor in neurite outgrowth from PC12D cells.

We isolated nardosinone as a neuritogenic substance from Nardostachys chinensis. Nardosinone did not exhibit the neurotrophic activity but caused a marked enhancement of the nerve growth factor (NGF)-mediated neurite outgrowth from PC12D cells. Nardosinone-induced enhancement of the NGF-action was completely blocked by PD98059, a representative mitogen activated protein kinase (MAPK) kinase inhibitor. The microscopic observations indicated that the neurites in response to nardosinone and NGF were quite long and were generally extended to the neighboring cells. These results suggest that nardosinone enhances the NGF-induced neurite outgrowth from PC12D cells probably by amplifying an up-stream step of MAPK kinase in the NGF receptor-mediated intracellular signaling pathway.

Animals↗

PubMed1999

Modulation of mammary development and programmed cell death by the frequency of milk removal in lactating goats.

1. Unilateral changes in mammary cell number are elicited when one gland is milked more or less frequently than the contralateral gland in lactating goats. These changes were investigated using histochemical and immunocytochemical markers of mammary cell types, and the degree of mammary apoptosis was determined by end-labelling of fragmented DNA. 2. Histological analysis confirmed that unilateral cessation of milking initiated involution and cell loss preferentially in the unmilked gland. The presence of fragmented DNA and morphological characteristics consistent with apoptosis demonstrated that these changes in mammary cell number in unmilked glands were, in part, the result of programmed alveolar cell death. 3. De-differentiation of the remaining secretory cells to ductal epithelial cells occurred with an increase in staining of cytokeratin markers and decreased staining by peanut lectin and casein antisera. 4. Differential once- and thrice-daily milking of lactating goats was also associated with unilateral changes in mammary cell number and milk yield. Milk yield and alveolar size were reduced after 4 weeks of infrequent milking. The latter was due to the increased loss of secretory cells by apoptosis, as indicated by a higher degree of fragmented DNA laddering. 5. After 10 weeks of differential milking, a homogeneous secretory morphology, albeit with smaller alveoli, was maintained in thrice-daily milked glands. Once-daily milked glands possessed a heterogeneous composition of terminal structures, resulting in the simultaneous presence of secretory and involuting alveoli as well as resting ductules. 6. The differences in programmed cell death and mammary morphology between unmilked and twice-daily milked glands, and between once- and thrice-daily milked glands, suggests that mammary apoptosis is subject to modulation by intra-mammary mechanisms sensitive to the frequency of milk removal.

Animals↗

PubMed1999

Molecular identification of the ryanodine receptor pore-forming segment.

A sequence motif, GXRXGGGXGD, located in the putative channel-forming domain, is conserved in all known ryanodine receptors and inositol 1,4,5-trisphosphate receptors. The functional significance of this conserved region was investigated by using site-directed mutagenesis together with functional assays consisting of Ca(2+) release measurements, [(3)H]ryanodine binding, and single channel recordings in planar lipid bilayers. We report here that single point mutations introduced into this region of the mouse cardiac ryanodine receptor reduce or abolish high affinity [(3)H]ryanodine binding. Single channel analysis revealed that a single substitution of alanine for glycine 4824 within this region reduced the single channel conductance by 97%, from 798 picosiemens (pS) for the wild type channel to 22 pS. The G4824A mutant channel was modulated by Ca(2+), Mg(2+), ATP, caffeine, ruthenium red, and ryanodine. Co-expression of the wild type and G4824A mutant proteins produced single channels that have intermediate unitary conductances of 516, 256, 176, and 60 pS. These data suggest that this conserved region constitutes an essential part of the ryanodine binding site and the channel conduction pathway of the ryanodine receptor.

Amino Acid Sequence↗

PubMed1999

Telomerase activity: comparison between fine-needle aspiration and biopsy specimens for the detection of tumor cells.

BACKGROUND: The purpose of the current study was to determine telomerase activity as a sensitive biomarker for the detection of malignant cells in fine-needle aspiration (FNA) specimens. METHODS: FNA specimens with parallel samples of fresh tumor tissue were obtained from surgical specimens after surgical excision. Using a polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay, telomerase activity was determined systematically in FNA specimens (n = 21) and corresponding available tissue biopsy specimens (n = 16) containing malignant cells. In addition to a case of myelolipoma, normal counterparts for 3 of 16 cancer cases, including both biopsy and FNA specimens, also were available for the determination of telomerase activity. RESULTS: Telomerase activity was observed in 14 of 16 of the FNA specimens (88%) and 15 of 16 of the corresponding biopsy specimens (94%). Telomerase activity was detected in both the biopsy specimen and the corresponding FNA specimen, with one exception (a case of mucinous adenocarcinoma of the cecum). In contrast, specimens from three normal tissue biopsies and FNA specimens of normal tissue adjacent to the malignant lesions, as well as the myelolipoma, exhibited no telomerase activity. It is interesting to note that both tissue biopsy specimens and FNA specimens from a patient with high grade sarcoma were negative for telomerase activity. The examination of hematoxylin and eosin-stained adjacent tissue biopsy sections or FNA smears revealed similar low populations of lymphocytes, including those cases that were negative for telomerase activity. There was agreement in the detection of telomerase activity between tissue biopsies and their corresponding FNA specimens in 15 of the 16 patients, indicating a 94% concordance rate (95% confidence interval, 70%, 98%). CONCLUSIONS: The results of the current study clearly suggest that the telomerase activity in FNA specimens was comparable to that of their corresponding biopsy specimens, and that this activity was associated with the presence of malignant cells. The TRAP assay has potential for use in the detection of malignant cells in FNA specimens, particularly cases in which the cytology is not characteristically malignant and/or is present in insufficient numbers. Cancer (Cancer Cytopathol)

Adolescent↗

PubMed1999

Zn2+ differentially modulates glycine receptors versus GABA receptors in isolated carp retinal third-order neurons.

It was previously reported that strychnine-sensitive glycine receptors and GABAA receptors co-existed on carp retinal third-order neurons (amacrine/ganglion cells) (Li, P. and Yang, X.-L., Strong synergism between GABAA and glycine receptors on isolated carp third-order neurons. NeuroReport, 9 (1998) 2785-2789.). In this study, the effects of the divalent cation Zn2+ on these two receptors were studied in amacrine/ganglion cells acutely isolated from carp, with the use of the whole-cell patch clamp recording technique. The glycine-induced currents were steadily potentiated by Zn2+ of lower concentrations (0.1-10 microM), while being dose-dependently inhibited by Zn2+ of higher concentrations ( > 100 microM). Both the effects involved changes in apparent glycine affinity of the glycine receptor. In contrast, Zn2+ consistently suppressed the GABA(A) mediated currents of these cells, and did not show similar dual effect. The differential modulation by Zn2+ of glycine receptors versus GABA(A) receptors provides a versatility for regulating inhibitory glycinergic and GABAergic inputs converging on most amacrine/ganglion cells.

Animals↗

PubMed1999

[Construction and characterization of non-replicating recombinant vaccinia virus expressing rabies glycoprotein].

OBJECTIVE: To improve the safety of recombinant vaccinia rabies virus, the non-replicating recombinant vaccinia rabies virus VTKRG delta CK was constructed. METHODS: The rabies glycoprotein(RG) gene was introduced into the TK locus of Chinese Tiantan strain of vaccinia virus (VTKRG), and a fragment between fragment C and fragment K involving vaccinia virus virulence and host-range related genes were deleted by homologous recombinant. RESULTS: The ability to replicate in primary chick chick embryo fibroblast(CEF) and dramatically reduced replicating capability in mammalian cells of the virus were observed, whereas VTKRG delta CK could express high level of RG efficiently. The expression of RG could be detected in CEF by indirect immunofluorescence assay and Western blot analysis. The VTKRG delta CK could elicit neutralizing antibody against CVS rabies virus and protect the animals from lethal rabies virus challenge in inoculated mice. CONCLUSION: VTKRG delta CK possesses immunogenicity and higher safety.

Antigens, Viral↗

PubMed1999

Regulatory features of the trp operon and the crystal structure of the trp RNA-binding attenuation protein from Bacillus stearothermophilus.

Characterization of both the cis and trans -acting regulatory elements indicates that the Bacillus stearothermophilustrp operon is regulated by an attenuation mechanism similar to that which controls the trp operon in Bacillus subtilis. Secondary structure predictions indicate that the leader region of the trp mRNA is capable of folding into terminator and anti- terminator RNA structures. B. stearothermophilus also encodes an RNA-binding protein with 77% sequence identity with the RNA-binding protein (TRAP) that regulates attenuation in B. subtilis. The X-ray structure of this protein has been determined in complex with L-tryptophan at 2.5 A resolution. Like the B. subtilis protein, B. stearothermophilus TRAP has 11 subunits arranged in a ring-like structure. The central cavities in these two structures have different sizes and opposite charge distributions, and packing within the B. stearothermophilus TRAP crystal form does not generate the head-to-head dimers seen in the B. subtilis protein, suggesting that neither of these properties is functionally important. However, the mode of L-tryptophan binding and the proposed RNA binding surfaces are similar, indicating that both proteins are activated by l -tryptophan and bind RNA in essentially the same way. As expected, the TRAP:RNA complex from B. stearothermophilus is significantly more thermostable than that from B. subtilis, with optimal binding occurring at 70 degrees C.

Amino Acid Sequence↗

PubMed1999

Blocking transforming growth factor beta signaling in transgenic epidermis accelerates chemical carcinogenesis: a mechanism associated with increased angiogenesis.

Mutations in the transforming growth factor beta type II receptor (TGF-betaRII) have been identified in human cancers, which suggests a causal role for the loss of TGF-betaRII in cancer development. To directly test this in vivo, we have generated transgenic mice expressing a dominant negative TGF-betaRII (delta betaRII) in the epidermis, using a truncated mouse loricrin promoter (ML). ML.delta betaRII transgenic mice exhibited a thickened skin due to epidermal hyperproliferation. When these mice were subjected to a standard two-stage chemical carcinogenesis protocol, they exhibited an increased sensitivity, with an earlier appearance and a 2-fold greater number of papillomas than control mice. In addition, papillomas in control mice regressed after termination of 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment; whereas ML.delta betaRII papillomas progressed to carcinomas. Furthermore, TPA promotion alone induced papilloma formation in ML.delta betaRII mice, which suggests an initiating role for delta betaRII in skin carcinogenesis. ML.delta betaRII tumors also exhibited increased neovascularization and progressed to metastases, although the primary tumors were still classified as carcinoma in situ or well-differentiated carcinomas. Increased expression of vascular endothelial growth factor, an angiogenesis factor, and decreased expression of thrombospondin-1, an angiogenesis inhibitor, were also observed in ML.delta betaRII tumors. The increased angiogenesis correlated with elevated endogenous TGF-beta1 in ML.delta betaRII tumors. These data provide in vivo evidence that inactivation of TGF-betaRII accelerates skin carcinogenesis at both earlier and later stages, and increased angiogenesis is one of the important mechanisms of accelerated tumor growth and metastasis.

Animals↗

PubMed1999

Transcribed dinucleotide microsatellites and their associated genes from channel catfish Ictalurus punctatus.

The presence of trinucleotide microsatellites within genes is a well-known cause for a number of genetic diseases. However, the precise distribution of dinucleotide microsatellites within genes is less well documented. Here we report 15 unique cDNAs containing dinucleotide repeats from the channel catfish Ictalurus punctatus. Gene identities of nine of the 15 cDNAs were determined, of which three encode structural genes, and six encode regulatory proteins. Five cDNAs harbored dinucleotide repeats in the 5' untranslated region (5'-NTR), nine in the 3'-NTR, and one in the coding region. The presence of these transcribed dinucleotide repeats and their potential expansion in size within coding regions could lead to disruption of the original protein and/or formation of new genes by frame shift. The low number of dinucleotide repeats within coding regions suggests that they were strongly selected against. All the transcribed microsatellite loci examined were polymorphic making them useful for gene mapping in catfish.

Animals↗