Biomedical subjects

L Yan

Publications and source records attributed to L Yan.

At least 73 records · Page 4Linked to original sources

PubMed2001

[Etiological study on human rotavirus infections in children with acute gastroenteritis].

OBJECTIVE: To understand the etiology of human rotavirus infections in children with acute gastroenteritis in China. METHODS: The investigation covered 1968 feces infants and children with acute gastroenteritis in 19 provinces and cities in China. Feces samples collected from 1968 infants and children with diarrhea, 148 children, 135 newborns and 37 adults without diarrhea, 36 adults with diarrhea were examined by polyacrylamide gel electrophoresis (PAGE). 1493 feces samples were examined by ELISA; 388 feces samples from sick children and 57 feces samples from children and adults without diarrhea were examined by EM. Bacterial culture were analyzed in 645 feces samples. RESULTS: Human rotavirus RNA (HRV RNA) was detected in 804(40.9%) of 1968 feces samples examined. No virus was detected in the feces samples of children and adults without diarrhea. Rotavirus antigen was detected in 500 (33.5%) of the 1493 feces samples. According to the genome profile of PAGE, 801 (99.6%) of the 804 rotavirus RNA positive belonged to group A virus, and the other 3 (0.4%) belonged to group C virus. According to the different migration of 10 and 11 segments on PAGE, the RNA pattern of group A rotavirus could be subdivided into two patterns: long and short patterns. The 527 (66.8%) positive HRV RNA was in the long pattern, while the 267 (33.3%) positive were in the short pattern. Among the rest 6 appeared to be mixed types, and the examined one was uncertain. Four different genotypes of HRV RNA in long and short patterns were identified respectively. They were types 4232, 4222, 3232 and 3222. The eight electropherotype genomes of HRV RNA varied not only in different years or seasons but also with the ages of patients and regions where the patients live. CONCLUSION: Rotavirus infections are the major cause of diarrhea of infants and young children throughout China.

Acute Disease↗

PubMed2001

[Prospective study of lethal blood concentration of organophosphorous in humans].

Total cases of organophosphorous (dichlorvos, methamidophos, dimethoate) poisoning outpatients from six hospitals during four years were collected consecutively for lethal blood concentration study. Blood samples were detected with gas chromatography. The probabilities of death, coma were analyzed with Bliss method and their linear regressive equations of probit were obtained respectively, their 50% lethal concentrations (LC50) and 50% coma concentrations(CC50) were calculated by the formulas above. As the death rate was influenced by therapy, its natural death probability has been discussed and estimated their natural LC50 were between the LC50 and CC50 themselves. Combined LC50 and CC50, their natural LC50 were calculated.

Adolescent↗

PubMed2001

[Study on the chemosensitivity test of human gastric cancer using the MTT assay].

OBJECTIVE: This study was conducted to investigate the chemosensitivity of gastric carcinoma. METHODS: 112 cases of fresh human gastric cancer samples were obtained for drug sensitivity testing using MTT. RESULTS: The sensitivity sequence of antitumor agents was 5-Fu, DDP, MMC, etc. On the other hand, the sensitivity was relative to pathology and individual differences. CONCLUSION: The MTT assay is an accurate system for determination of the chemosensitivity of tumor cells.

Adenocarcinoma↗

PubMed2000

Sum-frequency vibrational spectroscopy on chiral liquids: a novel technique to probe molecular chirality.

Optical activity in sum-frequency vibrational spectra has been observed for the first time in chiral liquids. The electric-dipole allowed chiral element of the nonlinear susceptibility appears to be 3 orders of magnitude smaller than typical allowed achiral elements. This is partly because the observed chirality requires a breakdown of the Born-Oppenheimer approximation.

Algorithms↗

PubMed2000

Phase-dependent responses of Per1 and Per2 genes to a light-stimulus in the suprachiasmatic nucleus of the rat.

Single brief and discrete light treatments are sufficient to reset the overt mammalian rhythms of nocturnal rodents. In the present study, we examined the phase-dependent response of the mammalian clock genes, Per1 and Per2, to a brief strong light-stimulus (1000 lux) in the circadian oscillator center, the suprachiasmatic nucleus (SCN) of rats. Light-induced elevation of Per1 mRNA was observed through the subjective night (CT16, CT20 and CT0 (=CT24)) with a marked peak at the subjective dawn (CT0). However, the light influence was very limited for the induction of Per2; only weak elevation of Per2 mRNA was detected at CT16. The effect of light-stimulus on the Per1 gene was transient, and the effect was restricted to ventrolateral SCN neurons in both CT0 and CT16 after light exposure. Since it is known that these rats show a light-induced behavioral phase-shift throughout the subjective night with being strongest at subjective dawn, the present results suggest that the transient induction of Per1 in ventrolateral SCN neurons is a critical step in the resetting of the biological clock to environmental light-dark schedule.

Animals↗

PubMed2000

The spider toxin omega-Aga IIIA defines a high affinity site on neuronal high voltage-activated calcium channels.

The spider toxin omega-agatoxin IIIA (omega-Aga-IIIA) is a potent inhibitor of high voltage-activated calcium currents in the mammalian brain. To establish the biochemical parameters governing its action, we radiolabeled the toxin and examined its binding to native and recombinant calcium channels. In experiments with purified rat synaptosomal membranes, both kinetic and equilibrium data demonstrate one-to-one binding of omega-Aga-IIIA to a single population of high affinity sites, with K(d) = approximately 9 pm and B(max) = approximately 1.4 pmol/mg protein. Partial inhibition of omega-Aga-IIIA binding by omega-conotoxins GVIA, MVIIA, and MVIIC identifies N and P/Q channels as components of this population. omega-Aga-IIIA binds to recombinant alpha(1B) and alpha(1E) calcium channels with a similar high affinity (K(d) = approximately 5-9 pm) in apparent one-to-one fashion. Results from recombinant alpha(1B) binding experiments demonstrate virtually identical B(max) values for omega-Aga-IIIA and omega-conotoxin MVIIA, providing further evidence for a one-to-one stoichiometry of agatoxin binding to calcium channels. The combined evidence suggests that omega-Aga-IIIA defines a unique, high affinity binding site on N-, P/Q-, and R-type calcium channels.

Agatoxins↗

PubMed2000

The 5' upstream region of mPer1 gene contains two promoters and is responsible for circadian oscillation.

The mPer1 gene is assumed to be a key molecule in the regulation and functioning of the mammalian circadian clock, which is based on the oscillation generated by a transcription-(post)translation feedback loop of a set of clock genes [1]. Robust circadian oscillation and acute light-elicited induction of mPer1 mRNA expression have been observed in the suprachiasmatic nucleus (SCN), the mammalian circadian center [2] [3]. To investigate the mechanism underlying the complex regulation of mPer1 expression, we isolated and characterized the 5' upstream region of the mPer1 gene. Unexpectedly, we identified two promoters, each followed by alternative first exons of mPer1. Consistent with the presence of multiple E-boxes in the promoters, exon-specific in situ hybridization of the SCN established that both promoters function in circadian oscillation and in light-induction of mPer1 expression. Transgenic mice carrying the 5' upstream region of the mPer1 gene fused to the luciferase gene demonstrated that a DNA fragment carrying both promoter regions is sufficient to elicit striking circadian oscillation in the SCN and responsiveness to light. Moreover, luminescence in the SCN accurately mirrored the mPer1 transcriptional activity. These transgenic mice will be very useful for monitoring clock-specific mPer1 expression in intact organisms and to follow the circadian clock in real time.

Animals↗

PubMed2000

Histamine N-methyltransferase functional polymorphism: lack of association with schizophrenia.

Histamine is a central nervous system (CNS) neurotransmitter that has been implicated in the pathophysiology of schizophrenia. Histamine N-methyltransferase (HNMT) terminates the neurotransmitter actions of histamine in the mammalian CNS, and levels of HNMT activity in human tissues are controlled, in part, by inheritance. A common C314T polymorphism in the HNMT gene causes a Thr105Ile change in encoded amino acid. The T314 allele results in decreased levels of both HNMT enzyme activity and immunoreactive protein. There is also a polymorphic CA repeat in intron 5 of the HNMT gene. The frequencies of alleles for the functional C314T polymorphism and the polymorphic CA repeat were compared between 171 schizophrenia cases and 171 ethnically matched controls to test for possible disease association. No significant difference was found between the two groups in the frequency of the T314 allele in patients with schizophrenia and controls (0.068 vs. 0.078, respectively). Allele frequencies for the polymorphic HNMT CA repeat also failed to show significant differences between cases and matched controls.

Alleles↗

PubMed2000

Superoxide production and reactive oxygen species signaling by endothelial nitric-oxide synthase.

Reactive oxygen species can function as intracellular messengers, but linking these signaling events with specific enzymes has been difficult. Purified endothelial nitric-oxide synthase (eNOS) can generate superoxide (O(2)) under special conditions but is only known to participate in cell signaling through NO. Here we show that eNOS regulates tumor necrosis factor alpha (TNFalpha) through a mechanism dependent on the production of O(2) and completely independent of NO. Expression of eNOS in transfected U937 cells increased phorbol 12-myristate 13-acetate-induced TNFalpha promoter activity and TNFalpha production. N(omega)-Methyl-l-arginine, an inhibitor of eNOS that blocks NO production but not its NADPH oxidase activity, did not prevent TNFalpha up-regulation. Likewise, Gln(361)eNOS, a competent NADPH oxidase that lacks NOS activity, retained the ability to increase TNFalpha. Similar to the effect of eNOS, a O(2) donor dose-dependently increased TNFalpha production in differentiated U937 cells. In contrast, cotransfection of superoxide dismutase with eNOS prevented TNFalpha up-regulation, as did partial deletion of the eNOS NADPH binding site, a mutation associated with loss of O(2) production. Thus, eNOS may straddle a bifurcating pathway that can lead to the formation of either NO or O(2), interrelated but often opposing free radical messengers. This arrangement has possible implications for atherosclerosis and septic shock where endothelial dysfunction results from imbalances in NO and O(2) production.

Binding Sites↗

PubMed2000

Cloning and characterization of DIP1, a novel protein that is related to the Id family of proteins.

Using human cyclin D1 as the "bait" in a yeast two-hybrid system, together with a HL60 cDNA library, we identified a novel human nuclear protein designated DIP1. This protein is expressed in a variety of cell types, and in fibroblasts its level remains constant throughout the cell cycle. However, the level of this protein increases severalfold during the differentiation of HL60 cells. The DIP1 protein can be phosphorylated in vitro by a cellular kinase and this activity reaches its maximum in extracts obtained from cells in the G1 phase of the cell cycle. DIP1 contains a helix-loop-helix motif but lacks an adjacent basic DNA-binding domain, thus resembling the Id family of proteins. The dip1 gene is located on human chromosome 16p11.2-12, a locus that is amplified in several types of human cancer. These results suggest that DIP1 may be involved in the control of gene expression and differentiation, but its precise function remains to be determined.

Amino Acid Sequence↗

PubMed2000

Matrix metalloproteinase-2 is required for the switch to the angiogenic phenotype in a tumor model.

Among the earliest and most important stages during tumorigenesis is the activation of the angiogenic process, an event that is termed the "switch to the angiogenic phenotype." We have developed an in vivo system that can reliably recapitulate the stages in tumor development that represent this transition. Using this model, we have harvested and studied tumor nodules that can be distinguished from each other on the basis of their degree of vascularization. Angiogenic tumor nodules were characterized by the presence of capillary vessels as determined by factor VIII immunohistochemistry, and both angiogenic and proteolytic activities in vitro. In contrast, preangiogenic nodules were devoid of microvessels and showed little angiogenic or proteolytic activity in vitro. Addition of a specific metalloproteinase inhibitor resulted in the abrogation of both angiogenic and proteolytic activities of the angiogenic nodules in vitro. Comparative substrate gel electrophoresis detected the presence of a prominent matrix metalloproteinase (MMP-2) in the angiogenic nodules when compared with the preangiogenic ones. Suppression of MMP-2 activity by antisense oligonucleotides in the vascular nodules resulted in the loss of angiogenic potential both in vitro and in vivo in the chick chorioallantoic membrane assay. Moreover, this suppression of MMP-2 activity in angiogenic nodules inhibited tumor growth in vivo by approximately 70%. These results strongly implicate the activity of MMP-2 as a requirement for the switch to the angiogenic phenotype and validate this model as a reliable and reproducible tool by which to study other cellular and biochemical factors involved in the acquisition of the angiogenic phenotype.

Animals↗

PubMed2000

Hydra metalloproteinase 1: a secreted astacin metalloproteinase whose apical axis expression is differentially regulated during head regeneration.

The newly emerging astacin metalloproteinase family comprises multiple members with diverse functions. Most recently, the development-related functions have been attributed to both (1) proteolytic cleavage and subsequent release of active TGF-beta-like growth factors from latent inhibitory complexes and (2) modification of extracellular matrix (ECM) assembly and composition. We previously identified and purified hydra metalloproteinase 1 (HMP-1), a developmentally important astacin proteinase that functions in head regeneration and transdifferentiation of tentacle battery cells (L. Yan et al., 1995, Development 121, 1591-1602). In the present study, further cloning revealed that HMP-1 is produced as a secreted zymogen with a conserved hydrophobic signal sequence and a putative propeptide. The processed HMP-1 is composed of a characteristic astacin proteinase domain and a unique Cys-rich C-terminus. With this simple domain structure, HMP-1 represents an ancestral astacin proteinase. Consistent with its role in head regeneration, HMP-1 mRNA is expressed at highest levels by endodermal cells at the apical pole of the body column just inferior to the base of tentacles, the region of active cell differentiation or transdifferentiation. A modified immunocytochemical procedure demonstrated that HMP-1 protein can be localized not only to ECM of tentacles as we previously reported, but also to endodermal cells of the body column in a pattern similar to its mRNA distribution. The localization of HMP-1 protein in tentacles was confirmed using an enzymatic approach. A translocation of HMP-1 protein from cells in the body column to the extracellular milieu in tentacles further suggests that HMP-1 is a secreted protein. HMP-1 expression undergoes extensive regulation at the transcriptional level both temporally and spatially during head regeneration. The involvement of HMP-1 in this morphogenetic process is further supported by the blockage of head regeneration with localized antisense treatment. Taken together, these results suggest that HMP-1 is a secreted astacin metalloproteinase that has an important role in regulating hydra head morphogenesis potentially through its differential expression along the body axis.

Amino Acid Sequence↗

PubMed2000

Distribution and circadian expression of dbp in SCN and extra-SCN areas in the mouse brain.

The expression of dbp, a putative clock-controlled transcription factor, was investigated in the mouse brain by in situ hybridization using antisense cRNA probe. Positive dbp mRNA signals were detected in various parts of the brain, with the highest expression in the suprachiasmatic nucleus (SCN). The circadian expression profile was investigated in SCN and extra-SCN areas. In the SCN, dbp mRNA signals showed a peak at early daytime (ZT/CT4) and a trough at early nighttime (ZT/CT16) in both light-dark and constant dark conditions. In the cerebral cortex and the caudate-putamen, dbp mRNA was also expressed in a circadian manner, but the phase of dbp mRNA expression in these structures showed a 4-8 hr delay compared to that in the SCN. These findings indicate that the circadian expression profile of dbp in the extra-SCN brain areas is different from that in the SCN.

Animals↗

PubMed2000

Identification and characterization of proSAAS, a granin-like neuroendocrine peptide precursor that inhibits prohormone processing.

Five novel peptides were identified in the brains of mice lacking active carboxypeptidase E, a neuropeptide-processing enzyme. These peptides are produced from a single precursor, termed proSAAS, which is present in human, mouse, and rat. ProSAAS mRNA is expressed primarily in brain and other neuroendocrine tissues (pituitary, adrenal, pancreas); within brain, the mRNA is broadly distributed among neurons. When expressed in AtT-20 cells, proSAAS is secreted via the regulated pathway and is also processed at paired-basic cleavage sites into smaller peptides. Overexpression of proSAAS in the AtT-20 cells substantially reduces the rate of processing of the endogenous prohormone proopiomelanocortin. Purified proSAAS inhibits prohormone convertase 1 activity with an IC(50) of 590 nM but does not inhibit prohormone convertase 2. Taken together, proSAAS may represent an endogenous inhibitor of prohormone convertase 1.

Adrenal Glands↗

PubMed2000

Prosaptide D5 reverses hyperalgesia: inhibition of calcium channels through a pertussis toxin-sensitive G-protein mechanism in the rat.

A retro-inverso 11-mer peptidomimetic of prosaposin, Prosaptide D5, induced neurite outgrowth in NS20Y neuroblastoma cells and enhanced [35S]GTPgammaS binding to rat synaptosomal membrane at low nanomolar concentrations similar to prosaposin. Intramuscular injection of D5 ameliorated thermal hyperalgesia in the Seltzer rat model of neuropathic pain, returning paw withdrawal latency to control levels within 3 h after treatment. The effect was sustained for at least 48 h after injection. Prosaposin and D5 inhibited K+-stimulated synaptosomal 45Ca2+ uptake similar to omega-conotoxin MVIIC, demonstrating that both effectors modulated voltage-dependent calcium channels (VDCC); inhibition was largely abolished by pretreatment with pertussis toxin before D5 treatment. The results suggest a mechanism whereby VDCC are modulated by a pertussis toxin-sensitive G-protein coupled receptor; D5 binds to this receptor and thereby ameliorates hyperalgesia in the Seltzer model of neuropathic pain.

Animals↗

PubMed2000

Three-dimensional model of human TIP30, a coactivator for HIV-1 Tat-activated transcription, and CC3, a protein associated with metastasis suppression.

Human TIP30 is a cofactor that specifically enhances human immunodeficiency virus-1 (HIV-1) Tat-activated transcription. The sequence of TIP30 is identical to that of CC3, a protein associated with metastasis suppression. TIP30/CC3 is a member of the short-chain dehydrogenases/reductases (SDR) family. Of the several experimentally determined SDR structures, Escherichia coli uridine diphosphate (UDP) galactose-4 epimerase is most similar to TIP30/CC3. Because the direct sequence similarity between TIP30/CC3 and E. coli UDP galactose-4 epimerase is low, we used the transitive nature of homology and employed two Aquifex aeolicus proteins as intermediaries in the homology modeling process. Comparison of our structural model with that of known SDRs reveals that TIP30/CC3 contains several well-conserved features, including a beta alpha beta fold at the amino terminus, which we predict binds NADP(H). TIP30/CC3 contains characteristic motifs at the catalytic site of SDRs, including a serine, tyrosine, and lysine that are important in catalyzing hydride transfer between substrate and cofactor. We also predict that a unique 20-amino acid sequence found at the amino terminus is an alpha-helix. Because this region contains several positively and negatively charged amino acids, it may dock TIP30/CC3 to other proteins. Our structural model points to this alpha-helix and the SDR-like part of TIP30/CC3 for mutagenesis experiments to elucidate its role in HIV-1 Tat-activated transcription, metastasis suppression, and other cellular functions.

Acetyltransferases↗

PubMed2000

A cnidarian homologue of translationally controlled tumor protein (P23/TCTP).

A protein homologous to P23, or translationally controlled tumor protein (TCTP), was cloned in Hydra vulgaris, the most ancient type of metazoan from which P23/TCTP has been characterized to date. Hydra P23/TCTP is composed of 184 amino acids and is encoded by a single mRNA of 700 bp. This invertebrate P23/TCTP is well conserved compared to those of other invertebrate and vertebrate species. Expression of Hydra P23/TCTP was confirmed by western blot of Hydra cell lysates using a polyclonal antibody against murine recombinant P23/TCTP. Spatial distribution of P23/TCTP mRNA and protein in Hydra was studied using in situ hybridization and immunostaining, respectively. Hydra P23/TCTP expression along the longitudinal body axis is regulated at both the transcriptional and the translational level. High levels of P23/TCTP mRNA were detected in a subpopulation of cells in the body column. In contrast, no mRNA was evident in the differentiated cells of the head and the foot regions. Coincidentally, P23/TCTP protein also concentrates to the body column, with no detectable protein in the head and foot region. However, despite the existence of P23/TCTP mRNA in both the ectoderm and endoderm in the body column, its protein is localized to the endodermal cells, suggesting a regulatory mechanism at the translational level. Taken together, the expression pattern of P23/TCTP in Hydra correlates with regions in which cell proliferation is actively occurring and its expression is excluded from regions where terminal differentiation has occurred.

Amino Acid Sequence↗

PubMed2000

Molecular and biological characterization of a zonula occludens-1 homologue in Hydra vulgaris, named HZO-1.

Zonula occludens-1 (ZO-1) is one of the earliest identified molecular components of tight junctions. Sequence analysis has placed ZO-1 into the broader membrane-associated guanylate kinase (MAGUK) protein family that contains such diverse members as post-synaptic density 95 (PSD-95), Drosophila discs large tumor suppressor gene product (dlg-A), p55, and TamA. Studies in both vertebrates and invertebrates have established that the MAGUK family is involved in a wide variety of cellular functions. These functions involve the regulation of such cellular processes as: (1) tight junction formation, (2) cell proliferation, (3) cell differentiation, and (4) neuronal synapse transmission. Extending these studies, we report the presence of a ZO-1 homologue in Hydra vulgaris, a member of the Cnidaria, the second oldest phylum of the animal kingdom. Hydra ZO-1 (HZO-1) is encoded by a single messenger RNA (mRNA) of approximately 6.0 kb that contains an open reading frame of 5,085 bp. The 191 kDa predicted protein consists of a characteristic MAGUK domain structure, including three PSD-95/SAP90, discs-large, ZO-1 (PDZ) domains, a src homology (SH3) domain, and a guanylate kinase (GUK) domain. Western blot analysis using an antibody generated from a synthetic peptide designed from the HZO-1 sequence confirmed the presence of a Hydra protein of the appropriate mass. While whole mount in situ hybridization determined that HZO-1 mRNA was expressed along the entire longitudinal axis of Hydra, cross-sectional analysis established that HZO-1 mRNA expression was restricted to the ectoderm or outer cell layer of the organism's epithelial bilayer. Consistent with this mRNA expression pattern, immunofluorescence studies localized HZO-1 protein to the apical plasma membrane of ectodermal cells. It is unclear what role HZ0-1 has in the cellular physiology of Hydra; however, immunolocalization studies indicate a conserved plasma membrane-associated function(s), as reported for its counterparts in other invertebrate and vertebrate species. These studies establish that the MAGUK family of proteins with a membrane-associated function arose early during metazoan evolution, even before the divergence of protostomes and deuterostomes.

Amino Acid Sequence↗