Biomedical subjects

X Liang

Publications and source records attributed to X Liang.

At least 217 records · Page 12Linked to original sources

PubMed1996

Cancer mortality among silicotics in China's metallurgical industry.

BACKGROUND: We conducted a retrospective cohort study of lung cancer among silicotics in China's metallurgical industries. The study cohort consisted of 4372 male silicotics diagnosed and alive before 1 January 1980. The follow-up period was 1 January 1980 to 31 December 1989. RESULTS: Standardized mortality ratios (SMR) of all deaths, all cancers, chronic bronchopneumonia, pneumonia and pulmonary heart disease, and pulmonary tuberculosis showed statistically significant excesses. The SMR for lung cancer for the total cohort was 237 (P < 0.01). When the total study cohort was divided into various categories by occupation (including iron ore mining, ore-sintering, refractory brick manufacturing, iron and steel smelting, and steel casting), significant excess lung cancer risk was observed among silicotics in all categories (SMR > 200, P < 0.01) except for casting (SMR = 157, P > 0.05). Excess lung cancer was not related to years since diagnosis of silicosis or years of exposure to silica. There was more than twofold excess lung cancer risk among both smoking (SMR = 257, P < 0.01) and non-smoking silicotics (SMR = 209, P < 0.01). Silicosis increased the risk of lung cancer.

China↗

PubMed1996

Bovine herpesvirus 1 UL49.5 homolog gene encodes a novel viral envelope protein that forms a disulfide-linked complex with a second virion structural protein.

We previously reported that the genome of bovine herpesvirus 1 (BHV-1) contains an open reading frame (ORF) homologous to the herpes simplex virus UL49.5 ORF, and as with the herpes simplex virus UL49.5 ORF, the deduced amino acid sequence of the BHV-1 UL49.5 homolog (UL49.5h) contains features characteristic of an integral membrane protein, implying that it may constitute a functional gene encoding a novel viral envelope protein. This communication reports on the identification of the BHV-1 UL49.5h gene product. By employing an antibody against a synthetic BHV-1 UL49.5h peptide and an UL49.5h gene deletion mutant, the primary product of BHV-UL49.5h gene was identified as a polypeptide with a size of approximately 9 kDa; in both infected cells and isolated virions, the UL49.5h products were found to exist in three forms; monomer, disulfide-linked homodimer, and disulfide-linked heterodimer containing a second viral protein with a size of about 39 kDa. O-Glycosidase digestion and [3H]glucosamine labelling experiments showed that the UL49.5h protein is not glycosylated. Although the deduced amino acid sequence contains putative sites for myristylation and phosphorylation, we were unable to detect either modification. Surface labelling and trypsin digestion protection experiments showed that the BHV-1 UL49.5h protein was present on the surface of infected cells and on the surface of mature virions. Nonionic detergent partition of isolated virions revealed that the UL49.5h protein is more tightly associated with the virion tegument-nucleocapsid structure than envelope protein gD. The results from this study demonstrate that the BHV-1 UL49.5h gene encodes a nonglycosylated virion envelope protein which may associate with virion internal structures by forming a complex with the 39-kDa virion structural protein.

Animals↗

PubMed1996

Glycoprotein Bb, the N-terminal subunit of bovine herpesvirus 1 gB, can bind to heparan sulfate on the surfaces of Madin-Darby bovine kidney cells.

The present study confirms our previous findings made by using heparin affinity chromatography that bovine herpesvirus 1 gB can bind to heparin-like structures. In order to locate the functional domain for heparin binding, we expressed the extracellular portion of gB (gBt) and the large subunit of gB (gBb) in Madin Darby bovine kidney (MDBK) cells under the control of the bovine heat shock protein 70A gene promoter. The recombinant gBt and gBb were both efficiently secreted from the transfected cells. They were shown to have structural and antigenic properties similar to those of authentic gB. Like authentic gB, both gBt and gBb were able to bind heparin-Sepharose as well as heparan sulfates on MDBK cells. Thus, we suggest that at least one heparin-binding domain is localized in gBb, the N-terminal portion of gB, which agrees with the presence of clusters of prolines and basic residues, thought to be essential for heparin binding.

Amino Acid Sequence↗

PubMed1996

Role of the C terminus of the glucocorticoid receptor in hormone binding and agonist/antagonist discrimination.

The glucocorticoid receptor (GR) is a hormone-inducible intracellular modulator of specific gene transcription. Both glucocorticoids and progestins bind to the GR, and some progestins are able to activate the receptor. We have characterized a mutation of the mouse GR that restricts transcriptional activation, but not hormone binding, to glucocorticoids. This mutation, Y77ON, is located 13 amino acids from the C terminus of the mouse GR and helps define a region of the receptor that is important for transcriptional specificity. To further characterize this region of the GR, we have constructed a series of chimeric receptors between the glucocorticoid, progesterone, and androgen receptors. We find that the C-terminal 14 amino acids of the GR can be replaced by the equivalent region of the progesterone or androgen receptors with little alteration in either hormone-binding specificity or transcriptional response to agonists and antagonists. The region is required for hormone binding, however, since C-terminal deletions yield inactive receptors. We conclude that even though mutation of the C-terminal 14 amino acids of the GR can lead to alterations in hormone binding specificity and agonist potential, the differential hormone-binding capacities of the glucocorticoid, progesterone, and androgen receptors are not encoded in this region.

Amino Acid Sequence↗

PubMed1996

[Photocoagulation and scleral cryotherapy for Coat's disease].

PURPOSE: To evaluate the results and significance of Coat's disease with argon laser photocoagulation and cryotherapy. METHODS: 22 patients (23 eyes) were treated by argon laser (German Opton Meditek 40 E, 0.2w power, 100-500 um spot, 0.1-0.2 second exposure and grade II reaction) and or scleral cryotherapy (-60 to approximately -80 degrees C, 5 second duration per time, repeating treatment if necessary until pale edema was seen in retina). RESULTS: 18 eyes in all (78.2%) had good results with visual acuity improvement and the lesion of fundus had been seen decreased in varient degrees. The follow-up ranged from 1 to 3 years. CONCLUSION: Photocoagulation and scleral cryotherapy are effective for those patients with Coat's disease. It is very important to treat them promptly so as to prevent the damage of visual function progressively.

Adolescent↗

PubMed1996

Morphological observations of vas deferens occlusion by the percutaneous injection of medical polyurethane.

Histological observations of 20 occluded vas segments obstructed by the standard procedure of medical polyurethane-vas occlusion in 10 volunteers requesting vasectomy and of 20 vas segments with plugs removed from 10 vas-occluded men requesting vas reversal were carried out in order to elucidate the mechanism of vas occlusion by medical polyurethane (MPU). Twenty vas deferens in vasectomy group were ruptured, only a small amount of MPU elastomer remained within the vas lumen, most having leaked through to encircle the ruptured vas. Histomorphology of the removed vas segments from 10 vas-occluded men, who had undergone MPU-vas occlusion 4 years earlier, showed diffuse proliferation of connective tissue, fibrosis, or hyalinization of fibroplastic tissue and local infiltration of lymphocytes and macrophages. The vas lumen both proximal and distal to the plugs was completely blocked. Sperm granuloma, foreign body granuloma, proliferation of nerve fibers and local infiltration of lymphocytes were found in four vas-occluded men with painful nodules. These results suggest that the contraceptive mechanism of MPU-vas occlusion could be the result of secondary obstruction due to tissue proliferation by MPU irritation after the rupture of the vas deferens.

China↗

PubMed1996

Novel, high expressing and antibiotic-controlled plasmid vectors designed for use in gene therapy.

The promise of effective gene therapy can only be accomplished by high-level expression and regulatable delivery of gene products. To achieve this end, a eukaryotic expression plasmid was modified to make transcription dependent on a tetracycline(Tc)-regulated chimeric transactivator. Mouse muscle injected with this two plasmid cis/trans control system expressed reporter proteins at levels five- to 10-fold greater than the cytomegalovirus immediate-early promoter-controlled parental plasmid. Tetracycline could be useful to either repress or activate transactivator-controlled expression based on the position of the tetO control sequences within the reporter plasmid. Finally, a prototype single plasmid construct was made and shown to express a self-regulating bicistronic transcript containing both the reporter and the transactivator. These Tc-controlled plasmids, termed maximum expression and regulated vectors (MERVs), have the potential to target a variety of gene therapy applications.

Animals↗

PubMed1996

[Changes in plasma endothelin-1 and lipid peroxidate levels and amount of superoxidate dismutase in red blood cell in patients with pregnancy-induced hypertension].

OBJECTIVE: To evaluate the role of endothelin (ET-1) and lipid peroxide (LPO) in the pathogenesis of pregnancy-induced hypertension (PIH). METHODS: The plasma concentrations of ET-1, LPO and the amount of superoxide dismutase (SOD) in red blood cell were measured by radioimmunoassay, thiobarbituric acid fluorimetric determination and catechol self-oxidation determination respectively in total 95 women (normal non-pregnant women 15, Late pregnant women 20 and patients with PIH 60). RESULTS: (1) The levels of ET-1, LPO and SOD in normal late pregnant women were significantly higher than those in non-pregnant women (P < 0.01), but the LPO/SOD ratio was not significantly different between the two groups. (2) The levels of plasma ET-1 and LPO/SOD ratio in cases with PIH were markedly higher than those in normal late pregnant women, and it increased with the severity of the disease and returned to the levels of normal late pregnant women within 3-7 days after delivery. (3) There was a positive correlation between ET-1 and LPO/SOD ratio in PIH. CONCLUSIONS: The imbalance of oxidation and antioxidation and endothelial cell injury may play an important role in the pathogenesis of PIH.

Adult↗

PubMed1996

[Experimental study on fitness of one-piece casting osseointegrated implant frameworks with relation to fixture abutment positions].

An experiment on fitness of one-piece casting osseointegrated implant frameworks was performed among three fixture abument positions where other conditions were controlled. The result revealed that there was statistically significant differences in dimensional change among the three fixture abument positions. In terms of the relation between abument position and fitness, the more backward the abument position was, the worse the osseointegrated framework fitness was. The frameworks showed deformations in "arch form" and "warp" in three dimensions.

Dental Abutments↗

PubMed1995

Characterization of two members (ACS1 and ACS3) of the 1-aminocyclopropane-1-carboxylate synthase gene family of Arabidopsis thaliana.

The nucleotide sequences of two highly homologous 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS; EC 4.4.1.14)-encoding genes, ACS1 and ACS3, of Arabidopsis thaliana (At) have been determined. The sequence analysis shows that ACS3 is a pseudogene representing a truncated version of ACS1. The missing region of ACS3 corresponding to the fourth exon of ACS1 has been shown by Southern analysis to be absent in the At genome. The chromosomal locations of the five members of the At ACS multigene family have been determined. The results show that each family member resides on a different chromosome. This observation suggests that the ACS3 pseudogene originated by a partial inter-chromosomal gene duplication. The ACS1 polypeptide contains all the conserved and characteristic domains found in the ACC synthase isoenzymes from various plant species, but is unable to express ACS activity in Escherichia coli and yeast. The predicted amino-acid sequence of ACS1 is missing the highly conserved tripeptide, Thr-Asn-Pro (TNP), between Ile204 and Ser205. Introduction of TNP into ACS1 restores the ACS activity, whereas its removal from the enzymatically active ACS2 results in a loss of activity. The results suggest that TNP is crucial for expression of ACS activity in E. coli.

Amino Acid Sequence↗

PubMed1995

Use of a nitrocellulose film substrate in matrix-assisted laser desorption/ionization mass spectrometry for DNA mapping and screening.

A nitrocellulose film is used as a substrate in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) studies of DNA. PCR products and DNA fragment digests obtained from biochemical procedures can be analyzed with the use of a nitrocellulose substrate in MALDI MS whereas no signal is observed with the use of a stainless steel substrate. In this method, on-probe purification allows for effective elimination of the interfering effects of salts, buffers, and other contaminants that are usually present in DNA samples, which serve as important limiting factors in the DNA molecular ion yield in the MALDI process. The use of the nitrocellulose film substrate also appears to improve the shot-to-shot and sample-to-sample reproducibility of the ion yield due to the more homogeneous coverage of matrix/analyte over the sample surface. With the use of the nitrocellulose substrate, DNA fragments of up to 622 base pairs in complex mixtures provide mass spectra with minimal sample purification. Although only species corresponding to single-stranded DNA were detected, a mass calibration scheme was established allowing an accuracy of within one base pair for fragments of < 250 bp under the experimental conditions. Despite the low mass resolution of the spectra obtained, the method has been successfully used for rapid DNA screening for sample disease genes and PCR products.

Base Sequence↗

PubMed1995

Delineation of the essential function of bovine herpesvirus 1 gD: an indication for the modulatory role of gD in virus entry.

The entry process of alphaherpesviruses consists of two steps, initial virus attachment and subsequent virus penetration involving membrane fusion. Glycoprotein D (gD) of the alphaherpesvirus bovine herpesvirus 1 (BHV 1) is an essential envelope protein, and it has been previously documented that gD plays a significant part in both of the virus entry steps. In order to gain further insight into the virus entry process, we attempted to define the essential function of BHV 1 gD. We replaced the gD transmembrane and cytoplasmic domains with a lipid-addition signal sequence from human decay accelerating factor and produced a stably transfected Madin Darby bovine kidney (MDBK) cell line that expresses a nonfusogenic, glycosylphosphatidylinositol (GPI)-anchored gD. We found that this cell line was able to support the growth of a gD gene-deletion mutant; the resultant gD mutant progeny contained the GPI-anchored gD on its virions and was able to enter into and produce a production infection in MDBK cells. This result suggests that fusion activity does not constitute the essential function of gD. In addition, we found that a gD-null virus (a virus containing no gD on its virion) could infect gD-expressing cells, but not normal MDBK cells. The ability of the gD-null virus to infect gD-expressing cells was dependent on the gD present on the cell surface, since either treating cells with phosphatidylinositol-specific phospholipase C to remove the GPI-anchored gD or incubating cells with gD monoclonal antibodies could block gD-null virus infection. This demonstrates that gD present on the cell surface can act in trans to facilitate the entry of virion lacking gD. This indicates that essential gD function can take place in the absence of gD-mediated virus attachment and membrane fusion. We also found that the gD monoclonal antibodies that block gD-null virus entry into gD-expressing cells are strictly restricted to the monoclonal antibodies that show postadsorption neutralization activity, indicating that the trans-acting function exhibited by the gD present on the cell surface represents the same function as defined by postadsorption antibody neurtralization. The results from this study suggest that the essential function of gD in virus entry is to modulate other virus-cell interaction(s) involved in productive virus penetration.

Amino Acid Sequence↗

PubMed1995

Rapid screening of genetic polymorphisms using buccal cell DNA with detection by matrix-assisted laser desorption/ionization mass spectrometry.

A new approach is developed for the rapid and cost-effective detection of human genetic polymorphisms based on matrix-assisted laser description/ionization mass spectrometric (MALDI MS) detection using a nitrocellulose film substrate. This method employs polymerase chain reaction (PCR) amplification using DNA extracted from buccal cells as templates, followed by direct digestion with restriction enzymes and subsequent analysis by MALDI MS. The extraction of DNA from buccal cells provides a rapid and convenient means for sampling PCR-based diagnostic analysis. The amount of DNA was sufficient as the template for both normal PCR amplifications, and amplifications involving the use of mismatched primers and multiple primers. The MALDI MS methodology has been successfully used for the analysis of such PCR products where restriction fragments generated directly in PCR reactions have been used for detection of carbonic anhydrase and cystic fibrosis transmembrane conductance regulator as model genes. The detection of genetic polymorphisms following routine biological and clinical procedures with the MALDI MS method is demonstrated. The results from MALDI MS analysis are shown to be comparable to those obtained from gel electrophoresis but the MALDI MS method is several orders of magnitude faster than gel electrophoretic techniques. The method described herein should also be readily extended to other areas involving DNA screening and testing.

Base Sequence↗

PubMed1995

Amiloride modulates urokinase gene expression at both transcription and post-transcription levels in human colon cancer cells.

Activity of receptor-bound urokinase plasminogen activator (uPA) on the surface of colon cancer cells appears to be a function of the number of uPA receptors. The regulation of uPA therefore may determine the invasive phenotype. The effects of amiloride on the modulation of uPA mRNA and protein induced by phorbol ester (PMA) and cycloheximide (CHX) were studied in four colon cancer cell lines, HCT116, KM12SM, LIM1215 and LS123. Northern blot analyses showed that PMA induced uPA mRNA that peaked at 2-48 h in HCT116 cells. In all colon cancer cell lines tested, the expression of uPA mRNA by PMA was super-induced after the addition of the protein synthesis inhibitor CHX, suggesting that stimulation of uPA gene expression does not require de novo protein synthesis. uPA mRNA was also induced by CHX alone, indicating that there may be a labile protein which inhibits uPA mRNA processing. Amiloride profoundly inhibited uPA mRNA production at concentrations between 0.1-1 mM in the presence or absence of PMA or CHX. uPA protein levels on the colon cancer cell surface reflected PMA induction and amiloride inhibition of uPA mRNA levels. Transcriptional elongation experiments using isolated nuclei indicated that while the induction effects of PMA or CHX on uPA gene expression were mediated at the post-transcriptional level, amiloride acted at both transcription and post-transcription levels. The inhibitory effects of amiloride on uPA gene expression reported in this paper may offer the prospect of developing new therapeutic approaches to the prevention of invasion and metastasis by adenocarcinomas.

Amiloride↗

PubMed1995

Adaptation of non-growing former epiphysis and metaphyseal trabecular bones to aging and immobilization in rat.

No data exit for distal tibia (DT), a bone site with growth plate closure at 3 months of age and with very low trabecular bone turnover rate. The purpose of this study was to characterize the response of DT to aging and immobilization (IM) in 6 to 16.5 month-old female rats. One hundred and forty six 6 month-old Sprague-Dawley rats were divided into aging controls and right hindlimb immobilization (RHLI) groups. The right hindlimb was immobilized against the abdomen with elastic bandages. Rats were sacrificed serially from 6 to 16.5 months old at intervals of 2 to 8 weeks. The histomorphometric study was performed on areas of former epiphyseal and metaphyseal trabecular bone of DT. No changes on bone mass or architecture were found in aging controls throughout the study. There was a decrease in bone mass of the former epiphysis after 3.5 months of IM due to resorption exceeding formation. Then it reverted to the aging control level at 4 months post IM. At the former metaphyseal site, significant bone loss (-19%) was observed as early as two weeks after IM. The peak bone loss (-54%) was reached at 1.5 months after IM and plateaued thereafter. The bone loss was mainly due to the reduction of trabecular thickness although trabecular number decreased significantly after 3.5 months IM. These results suggest that both former epiphysis and metaphyseal trabecular bones in DT can quickly adapt by losing bone to immobilization.

Adaptation, Physiological↗

PubMed1995

Characterization of bovine herpesvirus 1 UL49 homolog gene and product: bovine herpesvirus 1 UL49 homolog is dispensable for virus growth.

The sequence of the bovine herpesvirus 1 (BHV-1) gene that is homologous to the herpes simplex virus UL49 gene was determined. The BHV-1 UL49 homolog open reading frame consists of 774 bp and is capable of encoding 258 amino acids. Northern (RNA) blot analysis showed that the BHV-1 UL49 homolog is transcribed into a 1.1-kb RNA which is coterminal with the transcripts of an upstream UL49.5 homolog gene. Rabbit antisera produced against synthetic peptides of the predicted UL49 homolog gene product recognized a polypeptide of 33 to 35 kDa in both virus-infected cells and isolated virions. Further analysis by unionic-detergent partition of isolated virions suggested that the UL49 homolog gene product is a virion tegument protein. Indirect immunofluorescence assay revealed that the UL49 homolog gene product was predominantly localized in the nuclei of BHV-1-infected cells. A mutant virus with the UL49 homolog gene deleted was produced, and it was able to replicate in noncomplementing cells. Nevertheless, the yield of mutant virus was significantly reduced. The results from this study suggest that the BHV-1 UL49 homolog gene encodes a nuclear protein which constitutes a tegument component in mature virions and that it is dispensable for virus growth in cell culture.

Animals↗

PubMed1995

Characterization of cell-binding properties of bovine herpesvirus 1 glycoproteins B, C, and D: identification of a dual cell-binding function of gB.

Previous studies have suggested that the attachment of bovine herpesvirus 1 (BHV-1) to permissive cells is mediated by its major glycoproteins B (gB), C (gC), and D (gD). In order to gain further insight into the mechanism of the BHV-1 attachment process, we purified authentic gB, gC, and gD from BHV-1-infected cells and membrane anchor-truncated, soluble gB, gC, and gD from stably transfected cell lines by affinity chromatography and examined their cell-binding properties on Madin-Darby bovine kidney cells. All of the glycoproteins tested exhibited saturable binding to Madin-Darby bovine kidney cells. All of the glycoproteins tested exhibited saturable binding to Madin-Darby bovine kidney cells. Addition of exogenous heparin or treatment of cells with heparinase to remove cellular heparan sulfate (HS) prevented both gC and gB from binding to cells but had no effect on gD binding. An assessment of competition between gB, gC, and gD for cell binding revealed that gC was able to inhibit gB binding, whereas other combinations showed no effect. Cell-bound gC could be dissociated by heparin or heparinase treatment. The response of bound gB to heparin and heparinase treatments differed for the authentic and soluble forms; while soluble gB was susceptible to the treatment, a significant portion of cell-bound authentic gB was resistant to the treatment. Binding affinity analysis showed that soluble gB and both forms of gC and gD each had single binding kinetics with comparable dissociation constants (Kds), ranging from 1.5 x 10(-7) to 5.1 x 10(-7) M, whereas authentic gB exhibited dual binding kinetics with Kd1 = 5.2 x 10(-7) M and Kd2 = 4.1 x 10(-9) M. These results demonstrate that BHV-1 gC binds only to cellular HS, gD binds to a non-HS component, and gB initially binds to HS and then binds with high affinity to a non-HS receptor. Furthermore, we found that while authentic gB was able to inhibit viral plaque formation, soluble gB, which retains the HS-binding property but lacks the high-affinity binding property, was defective in this respect. These results suggest that the interaction between gB and its high-affinity receptor may play a critical role in the virus entry process.

Amino Acid Sequence↗

PubMed1995

Methods of training and constructing multilayer perceptrons with arbitrary pattern sets.

This paper presents two compensation methods for multilayer perceptrons (MLPs) which are very difficult to train by traditional Back Propagation (BP) methods. For MLPs trapped in local minima, compensating methods can correct the wrong outputs one by one using constructing techniques until all outputs are right, so that the MLPs can skip from the local minima to the global minima. A hidden neuron is added as compensation for a binary input three-layer perceptron trapped in a local minimum; and one or two hidden neurons are added as compensation for a real input three-layer perception. For a perceptron of more than three layers, the second hidden layer from behind will be temporarily treated as the input layer during compensation, hence the above methods can also be used. Examples are given.

Algorithms↗