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Biomedical subjects

X Feng

Publications and source records attributed to X Feng.

At least 163 records · Page 9Linked to original sources

Plasmid-mediated mineralization of carbofuran by Sphingomonas sp. strain CF06.

A bacterial strain (CF06) that mineralized both the carbonyl group and the aromatic ring of the insecticide carbofuran and that is capable of using carbofuran as a sole source of carbon and nitrogen was isolated from a soil in Washington state. Phospholipid fatty acid and 16S rRNA sequencing analysis indicate that CF06 is a Sphingomonas sp. CF06 contains five plasmids, at least some of which are required for metabolism of carbofuran. Loss of the plasmids induced by growth at 42 degrees C resulted in the inability of the cured strain to grow on carbofuran as a sole source of carbon. Introduction of the plasmids confers on Pseudomonas fluorescens M480R the ability to use carbofuran as a sole source of carbon for growth and energy. Of the five plasmids, four are rich in insertion sequence elements and contain large regions of overlap. Rearrangements, deletions, and loss of individual plasmids that resulted in the loss of the carbofuran-degrading phenotype were observed following introduction of Tn5.

Carbofuran↗

High- and low-abundance chemoreceptors in Escherichia coli: differential activities associated with closely related cytoplasmic domains.

In Escherichia coli, two high-abundance chemoreceptors are present in cellular dosages approximately ten-fold greater than two low-abundance receptors. In the absence of high-abundance receptors, cells exhibit an abnormally low tumble frequency and the ability of the remaining receptors to mediate directed migration in spatial gradients is substantially compromised. We found that increasing the cellular amount of the low-abundance receptor Trg over a range of dosages did not alleviate these defects and thus concluded that high- and low-abundance receptors are distinguished not simply by their different dosages in a wild-type cell but also by an inherent difference in activity. By creating hybrids of the low-abundance receptor Trg and the high-abundance receptor Tsr, we investigated the possibility that this inherent difference could be localized to a specific receptor domain and found that the cytoplasmic domain of the high-abundance receptor Tsr conferred the essential features of that receptor class on the low-abundance receptor Trg, even though it is in this domain that residue identity between the two receptors is substantially conserved.

Bacterial Proteins↗

[Study on the chemotaxis and DNA synthesis of pulmonary artery smooth muscle cells].

OBJECTIVE: To explore the effect of hypoxia on the proliferation and migration of pulmonary vascular smooth muscle cells (PVSMC), and whether migration of PASMC is involved in the pathogenesis of pulmonary vascular remodeling associated with hypoxia. METHODS: In this study, the effect of PDGF, ANP and hypoxia on DNA synthesis and chemotaxis of cultured neonatal calf pulmonary artery smooth muscle cells (PASMC) is investigated by 3H-thymidine incorporation and measurement of cell migration using a 48-well Boyden chamber respectively. RESULTS: The results demonstrated that hypoxia could stimulate DNA synthesis and chemotaxis of PASMC induced by PDGF; ANP could inhibit DNA synthesis and chemotaxis of PASMC by a cGMP dependent pathway. CONCLUSIONS: It is suggested that PDGF, ANP and hypoxia play important roles in regulating the proliferation and migration of PASMC, which is important in the pathogenesis of hypoxic pulmonary vascular remodeling.

Animals↗

Observation on electromyogram changes in 93 cases of peripheral facial paralysis treated by point-through-point acupuncture.

Bioelectricity of the affected muscles at acute stage, recovery stage and sequel state in 93 cases of peripheral facial paralysis was observed with EMG. Pathological potential or motor potential was found to be reduced to varying degrees in all the affected muscles. After treatment by point-through-point acupuncture, myodynamia recovered fairly rapidly, EMG showed obvious changes, pathological potential decreased, and the normal motor unit potential increased remarkably. Comparative analysis of EMG before and after acupuncture indicated that point-through-point acupuncture had an obvious therapeutic effect on the disease. It was also very significant that EMG was used in diagnosing various stages of peripheral facial paralysis, evaluation of prognosis and guidance in clinical treatment.

Acupuncture Points↗

[Alterations in metastatic properties of hepatocellular carcinoma cell following H-ras oncogene transfection].

OBJECTIVE: To demonstrate the relationship between H-ras oncogene and HCC metastasis. METHODS: Activated H-ras oncogenes were transfected into SMMC7721, a cell line derived from human hepatocellular carcinoma (HCC), by calcium phosphate transfection method. Some metastasis-related parameters were detected in vitro, including adhesion assay, migration assay, expresion of collagenase IV (cIVase) and EGFR. RESULTS: The abilities of H-ras-transfected cell clones in adhesion to laminin or fibronectin, migration, cIVase secretion increased markedly, and the expression of EGFR elerated moderately. More importantly, these alterations were consistently positive with the expression of p21, the protein product of H-ras oncogene. CONCLUSION: H-ras oncogene could induce the metastatic phenotype of HCC cell in vitro, therefore enhancing its metastatic potential.

Carcinoma, Hepatocellular↗

[Detection of desmosine in elastin by high performance liquid chromatography (HPLC)].

An assay of desmosine (DES) in elastin hydrolysate was performed by HPLC. Desmosine alone or its mixture with tyrosine and phenylalanine as well as elastin hydrolysate was separated on a C18 column eluted with a mobile phase of MeOH:H2O (2:8, V/V) and detected at UV-280nm. The results showed that the time of DES eluted in all cases mentioned above was comparable. The ratio of peak area (CA) and peak height (CH) between DES and Tyr is 1.4% and 3.4% resectively. It could be as a reliable measure to evaluate the quantity of DES in elastin. The correlation coefficient r was 0.9956 in the range of 0.16-0.64mg/L for a calibration curve of DES concentration vs peak area. In conclusion, this method could be used as a biochemical indication to detect the degradation level of elastin in the pathological state.

Chromatography, High Pressure Liquid↗

[Effects of vasoactive intestinal peptide, alpha-chymotrypsin, pancreozymin, lipase, phospholipase A2 and collagenase on the viscoelasticity properties of red blood cell suspension].

Using Low Shear-30 Rheometer, we studied the effects of vasoactive intestinal peptide, alpha-chymotrypsin, pancreozymin, lipase, phospholipase A2 and collagenase on the viscoelasticity properties of RBC suspension. The result showed that these drugs could increase the values of eta 0.512 and A. I. It suggests that these drugs could increase the degree of RBC aggregation. Among the drugs and concentrations, there is no significant difference.

Blood Viscosity↗

Isolation and sequencing of the gene encoding Sp23, a structural protein of spermatophore of the mealworm beetle, Tenebrio molitor.

The cDNA for Sp23, a structural protein of the spermatophore of Tenebrio molitor, had been previously cloned and characterized (Paesen, G.C., Schwartz, M.B., Peferoen, M., Weyda, F. and Happ, G.M. (1992a) Amino acid sequence of Sp23, a structure protein of the spermatophore of the mealworm beetle, Tenebrio molitor. J. Biol. Chem. 257, 18852-18857). Using the labeled cDNA for Sp23 as a probe to screen a library of genomic DNA from Tenebrio molitor, we isolated a genomic clone for Sp23. A 5373-base pair (bp) restriction fragment containing the Sp23 gene was sequenced. The coding region is separated by a 55-bp intron which is located close to the translation start site. Three putative ecdysone response elements (EcRE) are identified in the 5' flanking region of the Sp23 gene. Comparison of the flanking regions of the Sp23 gene with those of the D-protein gene expressed in the accessory glands of Tenebrio reveals similar sequences present in the flanking regions of the two genes. The genomic organization of the coding region of the Sp23 gene shares similarities with that of the D-protein gene, three Drosophila accessory gland genes and two Drosophila 20-OH ecdysone-responsive genes.

Amino Acid Sequence↗

Receptor-associated Mad homologues synergize as effectors of the TGF-beta response.

Transforming growth factor-beta TGF-beta is the prototype for a family of extracellular proteins that affect cell proliferation and tissue differentiation. TGF-beta-related factors, including BMP-2/4, Dpp and activin, act through two types of serine/threonine kinase receptors which can form a heteromeric complex. However, the mechanism of signal transduction by these receptors is largely unknown. In Drosophila, Mad is required for signalling by Dpp. We have isolated complementary DNAs for four human Mad homologues, one of which, hMAD-4, is identical to DPC-4, a candidate tumour suppressor. hMAD-3 and -4 synergized to induce strong ligand-independent TGF-beta-like responses. When truncated at their carboxy termini, hMAD-3 and -4 act as dominant-negative inhibitors of the normal TGF-beta response. The activity of hMAD-3 and -4 was regulated by the TGF-beta receptors, and hMAD-3 but not hMAD-4 was phosphorylated and associated with the ligand-bound receptor complex. These results define hMAD-3 and -4 as effectors of the TGF-beta response and demonstrate a function for DPCA-4/hMAD-4 as a tumour suppressor.

Activin Receptors, Type I↗

Competition for a unique response element mediates retinoic acid inhibition of vitamin D3-stimulated transcription.

We have identified a novel steroid hormone response element in the avian beta3 integrin promoter. This sequence, comprising three hexameric direct repeat half-sites separated by nine and three nucleotides binds vitamin D receptor (VDR)-retinoid X receptor (RXR) and retinoic acid receptor (RAR)-RXR heterodimers. VDR-RXR binds direct repeats separated by three base pairs, and RAR-RXR recognizes half-sites separated by nine bases, whereas the central half-site interacts with both heterodimers. Retinoic acid and 1, 25-dihydroxyvitamin D3 activate both a genomic fragment including the transcriptional start site and an oligonucleotide containing the three repeats, linked to a heterologous promoter. Co-addition of the steroids produces neither synergy nor an additive effect; rather the result equals that for retinoic acid alone. Scatchard analysis demonstrates that RAR-RXR has greater affinity than VDR-RXR for the composite element. Based on these findings we propose a model in which there is specific, polarity-defined binding of VDR-RXR and RAR-RXR to three half-sites, which form two overlapping steroid response elements, with the central half-site common to both. Our results identify a novel mechanism by which one steroid hormone can modulate the activity of a second, by competing for a shared half-site in a composite response element.

Animals↗

Structure of a D-protein gene and amino-acid sequences of the highly repetitive D-proteins secreted by the accessory glands of the mealworm beetle.

The D-group proteins form the major component of the proteinaceous secretion of the tubular accessory glands of the yellow mealworm beetle, Tenebrio molitor. In a previous paper, we reported the sequence of two D-protein cDNAs and their inferred translation products. Both proteins contain three highly repetitive domains (A, A' and B). In this paper, we present the cDNA-inferred sequences of 8 more D-proteins, none of which contains an A' domain. We also present the structure of a D-protein gene. Southern analysis suggests that genes coding for an A' domain are relatively rare. Genes with a total of 7 or 8 (A + B domain) repeats seem most common.

Amino Acid Sequence↗

A comparative study of glioma cell lines for p16, p15, p53 and p21 gene alterations.

A total of 10 glioma cell lines were examined for alterations of the p16, p15, p53 and p21 genes, which are tumor suppressor genes or candidates with direct or indirect CDK-inhibitory functions. Genetic alterations (deletions or mutations) were frequently seen in the p16, p15 and p53 genes in these cell lines, but not in the p21 gene. When the states of the p16, p15 and p53 genes were compared among cell lines, all the cell lines showed abnormalities in at least 1 gene, often in 2 or 3 genes coincidentally, suggesting that dysfunction of these genes is closely related to glioma cell growth. Although alteration of all 3 genes was most frequent, there were cell lines having either p16/p15 or p53 or pl6 and p53 gene alterations, suggesting that the time order of these genetic alterations was variable depending on the cell line. Among cell lines examined, one with homozygous p53 gene deletion seemed of particular practical value, since such a cell line might be useful in various studies, including investigation of the functions of various mutant p53 genes in the absence of heteromeric protein formation. On examination of the primary tumor tissues, the same alterations of the p16/p15 and p53 genes as detected in the cell lines were demonstrated in all 6 cases examined: p16/p15 gene deletion in 1, p16 gene mutation in 1 and p53 gene mutations in 5 cases. This suggested that the p16/p15 and the p53 gene alterations and their combinations in at least some glioma cell lines reflected those in the primary glioma tissues.

Adult↗

In vivo inhibition of hepatitis B viral gene expression by antisense phosphorothioate oligodeoxynucleotides in athymic nude mice.

Antisense oligodeoxynucleotides strategies have been used both to study normal gene function and to block gene expression therapeutically. We have previously shown that a number of antisense oligonucleotides against hepatitis B virus (HBV) mRNA are able to inhibit viral gene expression in vitro. Here we report the establishment of an animal model producing HBV markers in athymic nude mice and inhibition of HBV gene expression and replication by antisense DNA in vivo. 2.2.15 cells (Hep-G2 cell line transfected with HBV genomes) were injected subcutaneously (s.c.) into athymic BALB/c nude mice at a total cell number of 0.5-1 x 10(8) per mouse. Transplanted tumours developed about 2 weeks after inoculation. Hepatitis B surface and e antigens (HBsAg and HBeAg), as well as HBV DNA, could be detected in the circulation of tumour-bearing mice. Hepatitis B surface antigen and hepatitis B core antigen (HBcAg) were demonstrated in tumour cells. After 10 days of tumour growth, antisense phosphorothioate oligonucleotide, complementary to the cap site of the SP II promoter of HBV mRNA, were injected by infiltration into or around the tumour as a daily dose of 20 micrograms per gram body weight. Treatment for a total of 10 days resulted in an effective inhibition of viral replication and gene expression. These results suggest therapeutic potential for antisense oligomers in the treatment of patients who are chronically infected with HBV.

Animals↗

Dot-immunogold filtration assay as a screening test for syphilis.

A dot-immunogold filtration assay (DIGFA) for the rapid detection of reaginic antibody in the serum of syphilitic patients was developed. The assay was simple, rapid, and reproducible. The test completion time was 2 min, and the assay required no equipment. The positive dot was very obvious, and the results could easily be determined with the naked eye. A total of 350 serum samples were examined by DIGFA, the rapid plasma reagin test, and the fluorescent treponemal antibody-absorption test. The levels of agreement between DIGFA and the rapid reagin test and between DIGFA and the fluorescent treponemal antibody-absorption test were 100 and 98%, respectively. The results of clinical application indicated that DIGFA could be used as a routine screening test for syphilis.

Antibodies, Bacterial↗

Expression and function of the equine herpesvirus 1 virion-associated host shutoff homolog.

The ability of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2, respectively) to repress host cell protein synthesis early in infection has been studied extensively and found to involve the activities of the UL41 gene product, the virion-associated host shutoff (vhs) protein. To date, UL41 homologs have been identified in the genomes of three other alphaherpesviruses: equine herpesvirus 1 (EHV-1), varicella-zoster virus, and pseudorabies virus, but very little is known about the putative products of these homologous genes. Our earlier observations that no rapid early host protein shutoff occurred in EHV-1-infected cells led us to test EHV-1 vhs activity more thoroughly and to examine the expression and function of the EHV-1 UL41 homolog, ORF19. In the present study, the effects of EHV-1 and HSV-1 infections on cellular protein synthesis and mRNA degradation were compared at various multiplicities of infection in several cell types under an actinomycin D block. No virion-associated inhibition of cellular protein synthesis or vhs-induced cellular mRNA degradation was detected in cells infected with any of three EHV-1 strains (Ab4, KyA, and KyD) at multiplicities of infection at which HSV-1 strain F exhibited maximal vhs activity. However, further analyses revealed that (i) the EHV-1 vhs homolog gene, ORF19, was transcribed and translated into a 58-kDa protein in infected cells; (ii) the ORF19 protein was packaged into viral particles in amounts detectable in Western blots (immunoblots) with monoclonal antibodies; (iii) in cotransfection vhs activity assays, transiently-expressed ORF19 protein had intrinsic vhs activity comparable to that of wild-type HSV-1 vhs; and (iv) this intrinsic vhs activity was ablated by in vitro site-directed mutations in which either the functionally inactive HSV-1 vhs1 UL41 mutation (Thr at position 214 replaced by Ile [Thr-214-->Ile]) was recreated within ORF19 or two conserved residues within the putative poly(A) binding region of the ORF19 sequence were altered (Tyr-190, 192-->Phe). From these results we conclude that EHV-1's low vhs activity in infected cells is not a reflection of the ORF19 protein's intrinsic vhs activity but may be due instead to the amount of ORF19 protein associated with viral particles or to modulation of ORF19 protein's intrinsic activity by another viral component(s).

Amino Acid Sequence↗

Hypoxia and endothelin-1 stimulate DNA synthesis of pulmonary artery smooth muscle cells.

Hypoxia and endothelin-1 (ET-1) are associated with constriction of pulmonary vasculature both in vivo and in vitro. However, the role of hypoxia and ET-1 in the vascular remodelling during the development of pulmonary hypertension is unclear. This study demonstrated that ET-1 (0.1 nmol/L to 100 nmol/L) increased the [3H] thymidine uptake in a dose-dependent manner in cultured bovine pulmonary artery smooth muscle cells (PASMC), which was enhanced by exposing PASMC to hypoxia (2% O2, 93% N2, 5% CO2). BQ123, the specific antagonist of endothelin receptor subtype A, eliminated the ET-1 medicated proliferation of PASMC and the cooperative effect of hypoxia. Some dilatory drugs could inhibit the mitogenic effect of ET-1. We also observed that hypoxia significantly increased [3H]thymidine uptake in PASMC without ET-1 and BQ123 could inhibit this effect. Radioimmunoassay suggested that there was an autocrine of ET-1 in cultured PASMC which was enhanced by hypoxia significantly.

Animals↗

[Commodity identification of semen Plantaginis and herba Plantaginis].

The Semen Plantaginis from 29 provinces and regions and the Herba Plantaginis from 18 provinces and regions in our country were identified, and the main commodities of these two crude drugs are described in this paper. The identification may serve as a scientific basis for expanding new drug resources.

Drug Contamination↗