Biomedical subjects

L Xu

Publications and source records attributed to L Xu.

At least 685 records · Page 38Linked to original sources

PubMed1995

Interference of electromagnetic waves in dynamic metabolism.

Life is a continuous process of the dynamic metabolism. The influence of electromagnetic waves on the process of metabolism cannot be neglected. Here a new theory of electromagnetic interference in the dynamic metabolism of life is proposed. The statistical dynamic equations of ion and free radical in the biochemical reaction radiated by electromagnetic waves are given. The intensity of electromagnetic interference could be described with an interference factor. Good agreement can be seen between the calculated and measured results for a famous experiment of radio-frequency radiation-induced calcium ion efflux enhancement.

Animals↗

PubMed1995

Factors affecting removal and prognosis of thymic tumors.

One hundred and ten cases of thymic tumors were intervened surgically, including 92 thymoma, 8 thymic carcinoid, and 10 thymic carcinoma. In this series, 50.9% of the cases were complicated with various syndromes, 44.5% with myasthenia gravis (MG). Resection rate was correlated with the size and invasion of the tumor. There was significant difference in resection rate among thymoma, thymic carcinoid and thymic carcinoma. The degree of invasion undoubtedly influenced on resection. The 3-, 5- and 10- year survival rate of the thymoma were 82.7%, 68.1% and 40.0%, respectively. The prognosis depended on the pathological classification and the severity of the neighbouring invasion, but MG had no significant effect on prognosis. Recurrence and metastasis of the tumor were the main cause of late death.

Adult↗

PubMed1995

Characteristics of reversal optic cupping in adults glaucoma after reduction of intraocular pressure.

PURPOSE: According to Quigley's hypothesis the dense of connective tissue is the least and the pores of the lamina cribrosa are the largest at the superior and inferior poles of the lamina cribrosa, therefore they are the most vulnerable location to be involved in the characteristic glaucomatous optic nerve damage. To get clinical evidence for the hypothesis, the reversal of optic cup in adults glaucoma after reduction of intraocular pressure (IOP) was examined. METHODS: The stereoscopic flicker comparison on with computerized image system was used to monitor the optic cup's changing, 2 serial superposed stereo pairs were displayed alternatively and rapidly and the changing parts appeared moving Under the stereoscopic observation, the changes of 3-dimension optic cup could be seen and the false positive phenomena caused by photographic angle variation, vascular pulsation could be differentiated from the characteristic change of the cup. Stereo fundus photographs were taken from 31 eyes with hypertension glaucoma before and after treatment of reduction of IOPs which was either diamox administration or trabeculectomy. RESULTS: The result showed that the reversal of optic cups after reduction of IOPs were mostly asymmetrical, especially at the inferior and/or superior poles. By multiple stepwise regression, it is known that the amount of the change is only correlated with the initial elevated IOP. CONCLUSIONS: The study indicated that it was the distortion and deformity of the lamina cribrosa leading to shearing stress that results in glaucomatous characteristic damage in structure and function. The individual tolerance of the lamina cribrosa to the high intraocular pressure and the locations of individual vulnerable at the optic nerve head are various.

Adolescent↗

PubMed1994

Complete Nucleotide Sequence of Mouse Mammary Tumor Virus from JYG Chinese Wild Mice: Absence of Bacterial Insertion Sequences in the Cloned Viral gag Gene.

Mammary tumors of a newly isolated strain of Chinese wild mouse (JYG mouse) harbor exogenous mouse mammary tumor virus (MMTV). The complete nucleotide sequence of exogenous JYG-MMTV was determined on the proviral 5' long terminal repeat (LTR)(partial)-gag-pol-env-3' LTR (partial) fragment cloned into a plasmid vector and the cDNA sequence from JYG-MMTV producing cells. Similarly to the other MMTV species the LTR of JYG-MMTV contains an open reading frame (ORF). The amino acid sequence of the JYG-MMTV ORF resembles that of SW-MMTV (92% identity) and endogenous Mtv-7 (93% identity) especially at the C-terminal region. Thus, a functional similarity in T-cell receptor V beta recognition as a superantigen is implicated among these MMTV species. Analysis of the viral gag nucleotide sequence revealed that this gene is not disrupted by the bacterial insertion sequence IS1 or IS2, which have been reported to be present in the majority of the plasmids containing the gag region. Comparison of amino acid sequences of JYG-MMTV with those of BR6-MMTV showed that over 96% of the amino acids of gag, pol, protease and env products are identical. These results suggest the intact nature of the nucleotide sequence of the near full-length MMTV genome cloned in the plasmid.

Journal Article↗

PubMed1994

Nonviral and viral delivery of a human immunodeficiency virus protective gene into primary human T cells.

Because AIDS has been refractory to traditional pharmacologic interventions, alternative approaches have been developed. Although the introduction of specific antiviral genes into T leukemia cells can provide relative resistance to human immunodeficiency virus (HIV) replication, the testing of such genes against primary viral isolates in human CD4+ lymphocytes has been limited, and safety questions remain regarding gene delivery into cells from HIV-infected patients. In this report, we evaluate the efficacy of a transdominant mutant protein, Rev M10, against cloned and primary HIV isolates in human peripheral blood lymphocytes and describe different methods of gene transfer into peripheral blood lymphocytes from HIV-infected individuals. We show that gold microparticles can mediate stable Rev M10 gene transfer into these cells. Introduction of Rev M10 by these techniques conferred resistance to HIV infection in vitro to cloned and clinical isolates. Nonviral delivery of HIV protective genes will facilitate the development of gene therapy for AIDS and the analysis of viral and cellular gene expression in human T lymphocytes.

Base Sequence↗

PubMed1994

Epidemiological and genetic studies of congenital profound deafness in the general population of Sichuan, China.

People with congenital profound deafness (CPD) were surveyed in the general population of Sichuan. The prevalence was 0.082% (104/126, 876) of the general population, male 0.086%, female 0.078%. There was no significant difference in prevalence between urban and rural populations. However the population living in the mountains had a much higher prevalence than people from the foothills and plains (P < 0.05). Amongst the nationalities investigated, there were significant differences in prevalence. Non-Chinese, except for Tibetans, presented a significantly higher prevalence than Han Chinese. There was a significantly higher prevalence among the inbred population (0.82%) than among the non-inbred population (0.72%). An effect of parental age was demonstrated, but no effect of birth order was found. There was a significant seasonal variation in prevalence. Inherited cases could account for 71.2% of all cases, of which 92% were autosomal recessive (AR) and 8% autosomal dominant (AD). Heterogeneity in AR was found with at least 8 different loci. The fitness was 60.26%, the coefficient of selection was 0.3974, mutation rate was estimated to be 2.0 x 10(-4), and no heterozygote advantage was proven.

Adolescent↗

PubMed1994

TP53 gene mutations and CCND1 gene amplification in head and neck squamous cell carcinoma cell lines.

Mutations of the tumor-suppressor gene TP53 and amplification of CCND1 gene have been reported to occur frequently in head and neck squamous cell carcinomas (HNSQCC). In experimental systems, TP53 mutations have been shown to lead to genomic instability, including an increased propensity for gene amplification. We have examined 16 HNSQCC cell lines for the association between TP53 over-expression/mutation and CCND1 amplification. p53 over-expression was detected in 50% of the cell lines by immunohistochemistry using the monoclonal antibody (MAb) PAb1801. TP53 mutations were also detected in 50% of the cell lines by analysis of single-strand conformation polymorphism (SSCP) and DNA sequencing of exons 4 through 9. Six cell lines showed TP53 mutations and over-expression of the protein, 2 cell lines showed TP53 mutations but no p53 expression, and 2 cell lines showed over-expression of p53 protein but no TP53 gene mutations. CCND1 amplification was found in 38% of the cell lines by Southern blot analysis. Only 1 cell line showed both TP53 mutation and CCND1 amplification, whereas 7 of 8 cell lines with TP53 mutations had no CCND1 amplification. pRb expression was detected by Western blot analysis, and the level of pRb did not correlate with either CCND1 amplification or TP53 mutation. Our findings suggest that TP53 mutation and CCND1 amplification are common genetic alterations in HNSQCC and that the occurrence of either genetic event may be sufficient to abrogate normal cell cycle control.

Carcinoma, Squamous Cell↗

PubMed1994

Mutational analysis of CDKN2 (MTS1/p16ink4) in human breast carcinomas.

The CDKN2 gene that encodes the cell cycle regulatory protein cyclin-dependent kinase-4 inhibitor (p16) has recently been mapped to chromosome 9p21. Frequent homozygous deletions of this gene have been documented in cell lines derived from different types of tumors, including breast tumors, suggesting that CDKN2 is a tumor suppressor gene involved in a wide variety of human cancers. To determine the frequency of CDKN2 mutations in breast carcinomas, we screened 37 primary tumors and 5 established breast tumor cell lines by single-strand conformation polymorphism analysis. In addition, Southern blot analysis was performed on a set of five primary breast carcinoma samples and five breast tumor cell lines. Two of the five tumor cell lines revealed a homozygous deletion of the CDKN2 gene, but no mutations were observed in any of the primary breast carcinomas. These results suggest that the mutation of the CDKN2 gene may not be a critical genetic change in the formation of primary breast carcinoma.

Breast Neoplasms↗

PubMed1994

Cloning and expression of phosphatidylcholine-hydrolyzing phospholipase D from Ricinus communis L.

Phosphatidylcholine-hydrolyzing phospholipase D (PLD; EC 3.1.4.4) has been proposed to play an important role in the signal transduction pathways in animals and in various cellular processes in plants. To unravel the structure of PLD and further the investigation of its modes of regulation and cellular function, we have isolated a PLD cDNA from castor bean (Ricinus communis L. var. Hale) by using oligonucleotide probes based on the N-terminal amino acid sequence of purified PLD protein. The nucleotide sequence of the castor bean PLD cDNA clone contains an open reading frame of 2424 bases encoding an 808-amino acid protein of 92,400 daltons. Expression of this PLD cDNA clone in Escherichia coli resulted in the accumulation of a functional PLD able to catalyze hydrolysis and transphosphatidylation reactions, demonstrating that the introduction of this single gene product was sufficient to confer PLD activity and that both the hydrolysis and transphosphatidylation reactions are catalyzed the single PLD protein. Comparison of the deduced protein sequence of the clone to the N-terminal amino acid sequence of purified PLD revealed that the mature PLD protein is preceded by a 30-amino acid leader peptide. Removal of this leader peptide resulted in accumulation of non-functional PLD and also increased PLD degradation in E. coli, suggesting that this leader peptide may be involved in proper folding of PLD. The primary structure of the castor bean PLD exhibits no significant similarities with sequences of other cloned lipolytic enzymes.

Amino Acid Sequence↗

PubMed1994

Optic disk hemorrhages in glaucoma.

We evaluated the frequency and location of optic disk hemorrhages by examining morphometrically color stereo optic disk photographs of 1,505 glaucomatous eyes and 595 normal eyes. Disk hemorrhages were found in 61 of the glaucomatous eyes (4.1%) and in none of the normal eyes. The frequency of the hemorrhages increased from an early stage of glaucoma to a medium-advanced stage and decreased again toward a far-advanced stage. The disk hemorrhages were not found in disk regions or eyes without detectable neuroretinal rim. In early glaucoma, they were usually located in the inferotemporal or superior disk regions. (The frequency of hemorrhages was 6.3% in normal-pressure glaucoma, 4.9% in primary open-angle glaucoma, and 3.1% in secondary open-angle glaucoma). They were correlated with localized retinal nerve fiber layer defects, neuroretinal rim notches, and circumscribed perimetric loss. These characteristics imply the pathogenetic and diagnostic importance of disk hemorrhages for the detection of glaucoma.

Adolescent↗

PubMed1994

Suppression of Myc, but not E1a, transformation activity by Max-associated proteins, Mad and Mxi1.

Mad and Mxi1, two members of the Myc-related basic-region helix-loop-helix/leucine-zipper family of proteins, associate directly with Max to form sequence-specific DNA binding heterodimers that are transactivation-incompetent. Mad-Max complexes have been shown to exert a strong repressive effect on Myc-induced transactivation, perhaps through the competitive occupation of common promoter binding sites also recognized by active Myc-Max heterodimers. To place these recent biochemical observations in a biological context, mad and mxi1 expression vectors were tested for their ability to influence Myc transformation activity in the rat embryo fibroblast cooperation assay. Addition of an equimolar amount of mad or mxi1 expression vector to mouse c-myc/ras cotransfections resulted in a dramatic reduction in both the number of foci generated and the severity of the malignant phenotype. Myc-specific suppression by Mad and Mxi1 was demonstrated by their ability to affect c- and N-myc-, but not ela-, induced transformation. In contrast, mad and mxi1 expression constructs bearing deletions in the basic region exerted only mild repressive effects on Myc transformation activity, suggesting that occupation of common DNA binding sites by transactivation-incompetent Mad-Max or Mxi1-Max complexes appears to play a more dominant role in this suppression than titration of limited intracellular pools of Max away from active Myc-Max complexes. Thus, these biological data support a current model for regulation of Myc function in which relative intracellular levels of Mad and Mxi1 in comparison to those of Myc may determine the degree of activation of Myc-responsive growth pathways.

Adenovirus E1A Proteins↗

PubMed1994

Intrinsic molecular activities of the interferon-induced 2-5A-dependent RNase.

2-5A-dependent RNase (RNase L), a unique endoribonuclease that requires 5'-phosphorylated 2',5'-linked oligoadenylates (2-5A), functions in the molecular mechanism of interferon action. Because this enzyme is present at very low levels in nature, characterization and analysis have been limited. The molecular cloning of human, 2-5A-dependent RNase cDNA has facilitated its expression to high levels in insect cells by infecting with recombinant baculovirus. To determine the properties of the enzyme in the absence of other proteins, the recombinant 2-5A-dependent RNase was purified to homogeneity. The purified enzyme migrated as a monomer upon gel filtration in the absence of activator and showed highly specific, 2-5A-dependent RNase activity. The precise activator requirements were determined by stimulating the purified enzyme with a variety of 2',5'-linked oligonucleotides. The activated enzyme was capable of cleaving poly(rU) and, to a lesser extent, poly(rA), to sets of discrete products ranging from between 4 and 22 nucleotides in length. Reduced rates of 2-5A-dependent RNA cleavage were observed even after removal of ATP and chelation of divalent cations. However, optimal RNA cleavage rates required the presence of either manganese or magnesium and ATP.

Adenosine Triphosphate↗

PubMed1994

A panoramic code for sound location by cortical neurons.

By conventional spike count measures, auditory neurons in the cat's anterior ectosylvian sulcus cortical area are broadly tuned for the location of a sound source. Nevertheless, an artificial neural network was trained to classify the temporal spike patterns of single neurons according to sound location. The spike patterns of 73 percent of single neurons coded sound location with more than twice the chance level of accuracy, and spike patterns consistently carried more information than spike counts alone. In contrast to neurons that are sharply tuned for location, these neurons appear to encode sound locations throughout 360 degrees of azimuth.

Acoustic Stimulation↗

PubMed1994

Reconstitution of the skeletal muscle ryanodine receptor-Ca2+ release channel protein complex into proteoliposomes.

The 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid-solubilized 30 S ryanodine receptor (RyR)/Ca2+ release channel complex from rabbit skeletal muscles, purified by density gradient centrifugation, was reconstituted with an excess of phospholipid into proteoliposomes by removal of the detergent by dialysis. Reconstituted proteoliposomes were concentrated by centrifugation, frozen and thawed, and sonicated. [3H]Ryanodine binding measurements indicated close to 50% recovery of calculated binding activity following reconstitution of the purified RyR and dynamic light scattering measurements a mean vesicle diameter of approximately 150 nm. Using these values, a functional RyR was estimated to be present in only a small fraction (< 15%) of the reconstituted vesicles. SDS-polyacrylamide gel electrophoresis of trypsin-treated proteoliposomes revealed that about four-fifths of the reconstituted 30 S complex was readily accessible to proteolytic attack. Vesicle-45Ca2+ flux and fusion of proteoliposomes with planar lipid bilayers showed that the reconstituted channel complex could be activated by Ca2+ and ATP, inhibited by Mg2+ and ruthenium red, and modified by ryanodine, similarly as observed for native sarcoplasmic reticulum vesicles. These results suggest that the method described here results in the reconstitution of a functional channel and thus provides the opportunity to study the structure and function of the sarcoplasmic reticulum Ca2+ release channel under well defined conditions.

Animals↗

PubMed1994

Evidence for a Ca(2+)-gated ryanodine-sensitive Ca2+ release channel in visceral smooth muscle.

Although a role for the ryanodine receptor (RyR) in Ca2+ signaling in smooth muscle has been inferred, direct information on the biochemical and functional properties of the receptor has been largely lacking. Studies were thus carried out to purify and characterize the RyR in stomach smooth muscle cells from the toad Bufo marinus. Intracellular Ca2+ measurements with the Ca(2+)-sensitive fluorescent indicator fura-2 under voltage clamp indicated the presence of a caffeine- and ryanodine-sensitive internal store for Ca2+ in these cells. The (CHAPS)-solubilized, [3H]ryanodine-labeled RyR of toad smooth muscle was partially purified from microsomal membranes by rate density centrifugation as a 30-S protein complex. SDS/PAGE indicated the comigration of a high molecular weight polypeptide with the peak attributed to 30-S RyR, which had a mobility similar to the cardiac RyR and on immunoblots cross-reacted with a monoclonal antibody to the canine cardiac RyR. Following planar lipid bilayer reconstitution of 30-S stomach muscle RyR fractions, single-channel currents (830 pS with 250 mM K+ as the permeant ion) were observed that were activated by Ca2+ and modified by ryanodine. In vesicle-45Ca2+ efflux measurements, the toad channel was activated to a greater extent at 100-1000 microM than 1-10 microM Ca2+. These results suggest that toad stomach muscle contains a ryanodine-sensitive Ca2+ release channel with properties similar but not identical to those of the mammalian skeletal and cardiac Ca(2+)-release channels.

Animals↗

PubMed1994

Activation of the skeletal muscle calcium release channel by a cytoplasmic loop of the dihydropyridine receptor.

Expression studies with skeletal and cardiac muscle cDNAs have suggested that the putative cytoplasmic loop region of the dihydropyridine receptor (DHPR) alpha 1 subunit between transmembrane repeats II and III (DCL) is a major determinant of the type of excitation-contraction coupling (skeletal or cardiac) in rescued dysgenic muscle cells (Tanabe, T., Beam, K. G., Adams, B. A., Niidome, T., and Numa, S. (1990) Nature 346, 567-569). In this study, the possibility of a direct functional interaction with the sarcoplasmic reticulum ryanodine receptor/Ca2+ release channel has been tested by expressing the DCLs of the mammalian skeletal and cardiac muscle DHPR alpha 1 subunit in Escherichia coli. The purified peptides activated the skeletal muscle ryanodine receptor/Ca2+ release channel in single channel and [3H]ryanodine binding measurements, by increasing channel open probability and the affinity of [3H]ryanodine binding, respectively. The two peptides did not activate the cardiac muscle Ca2+ release channel. Other proteins (polylysine, serum albumin) also increased [3H]ryanodine binding and Ca2+ release channel activity, but their activation mechanisms were distinguishable from DCLs. These results show that the II-III cytoplasmic loop of the skeletal and cardiac DHPR alpha 1 subunit functionally interacts with the skeletal, but not cardiac, muscle Ca2+ release channel. Furthermore, our studies suggest that in addition to the DHPR, the sarcoplasmic reticulum Ca2+ release channel may determine the type of E-C coupling that exists in muscle.

Amino Acid Sequence↗

PubMed1994

Molecular characterization of the mouse ribosomal protein S24 multigene family: a uniquely expressed intron-containing gene with cell-specific expression of three alternatively spliced mRNAs.

A family of 16 genes encoding the mouse ribosomal protein S24 was identified, and four members from this family were cloned. A single expressed intron-containing S24 gene (termed mrpS24) and one pseudogene (mrpS24p) were completely sequenced and characterized. The mrpS24 gene has seven exons and six introns spanning over 5.1 x 10(3) nucleotides (nt). The cap site of S24 was mapped to a G residue four nt upstream of a polypyrimidine tract and 15 nt downstream of a TATA-like (TATGA) element. The 5' region (-325 to +33) of the mrpS24 gene has a functional promoter that was able to express the fused chloramphenicol acetyltransferase (CAT) reporter gene. Two different forms of mouse S24 cDNA clones were previously isolated. Sequence analysis showed that one of these cDNA clones (termed S24a) lacks the entire exon V sequence (18 nt), and the deduced amino acid sequence is missing a C-terminal lysine residue encoded by the other cDNA (S24b). The pseudogene mrpS24p is flanked by an 11-bp direct repeat, and its sequence is almost identical to the S24 cDNA sequence, but it lacks two mini-exons, V and VI (20 nt), as in the cases of the human and rat S24 cDNAs. RT-PCR experiments demonstrated the existence of a third form (S24c) that similarly lacks both of the mini-exons, and suggested that different species of S24 mRNA might arise from alternative splicing of the mini-exons V and VI. Northern blot analysis showed that S24 expression is down- and up-regulated during adipocyte differentiation and in cellular transformation, respectively. RNase protection assays and RT-PCR experiments suggested that these cell-specific changes of S24 mRNA levels are mainly due to fluctuations in S24c mRNA level. Our results provide the first indication that a ribosomal protein gene is regulated by alternative usage of two mini-exons in a cell-specific manner.

Alternative Splicing↗

PubMed1994

S region transcription per se promotes basal IgE class switch recombination but additional factors regulate the efficiency of the process.

Stimulation of B lymphocytes with a combination of lipopolysaccharide (LPS) and interleukin-4 (IL-4) induces germline transcription of and subsequent switching to the epsilon heavy chain constant region (C epsilon) gene. Mature germline C epsilon transcripts contain a non-coding exon (I epsilon exon) spliced to the C epsilon exons. To distinguish between the potential roles of germline transcription and those of germline transcripts in regulating the class switch process, we replaced the LPS- and IL-4-inducible I epsilon promoter and exon in ES cells with an LPS-inducible E mu enhancer/VH promoter expression cassette. Wildtype, heterozygous or homozygous mutant ES cells were injected into RAG-2 deficient blastocysts to generate somatic chimeras in which all B cells derived from ES cells. In contrast to normal B cells, heterozygous and homozygous mutant B cells had substantial transcription through the epsilon switch recombination region (S epsilon) following treatment with LPS alone and, under these conditions, both underwent low level switching (10- to 100-fold less than wildtype cells stimulated with LPS + IL-4) to IgE production. Heterozygous mutant cells underwent switching to IgE at essentially wildtype levels when stimulated with LPS and IL-4. However, homozygous mutant cells still showed extremely low levels of switching to IgE upon LPS and IL-4 stimulation. Analyses of hybridomas from heterozygous mutants indicated that the mutation is cis-acting and normal switching to other isotypes indicated that it is specific for IgE. Thus transcription per se generates low levels of class switch recombination in the absence of I region sequences. However, we demonstrate for the first time that, for optimal efficiency, the process requires the presence of the intact I region and/or I region promoter in cis, implicating factors beyond transcription through the S region in the regulation of class switching.

Animals↗