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Biomedical subjects

X Dong

Publications and source records attributed to X Dong.

At least 181 records · Page 10Linked to original sources

Creation of a high cytotoxic active human tumor necrosis factor having the truncated and more basic amino terminus.

In order to define the structure-functional relationship of tumor necrosis factor(TNF), a mutant TNF gene was created by site-specific mutagenesis based on the PCR technique. This gene was highly expressed in E.coli cells. The amount of the recombinant protein was up to about 80% of the total cellular proteins. Through one-step ion exchange chromatography, the mutant TNF could be purified to homogeneity. This mutein showed the molecular weight of a dimer but not a trimer. It bears the features of truncated amino terminus and increase of the basicity of amino terminal residues. Compared with the wild type TNF, the specific activity of mutant TNF was increased by fourfold.

Amino Acid Sequence↗

Localization of the enzymes involved in H2 and formate metabolism in Syntrophospora bryantii.

Cell-free extracts of crotonate-grown cells of the syntrophic butyrate-oxidizing bacterium Syntrophospora bryantii contained high hydrogenase activities (8.5-75.8 mumol.min-1mg-1 protein) and relatively low formate dehydrogenase activities (0.04-0.07 mumol.min-1 mg-1 protein). The KM value and threshold value of the hydrogenase for H2 were 0.21 mM and 18 microM, respectively, whereas the KM value and threshold value of the formate dehydrogenase for formate were 0.22 mM and 10 microM, respectively. Hydrogenase, butyryl-CoA dehydrogenase and 3-OH-butyryl-CoA dehydrogenase were detected in the cytoplasmic fraction. Formate dehydrogenase and CO2 reductase were membrane-bound, likely located at the outer aspect of the cytoplasmic membrane. Results suggest that during syntrophic butyrate oxidation H2 is formed intracellularly while formate is formed at the outside of the cell.

Aldehyde Oxidoreductases↗

Role of formate and hydrogen in the degradation of propionate and butyrate by defined suspended cocultures of acetogenic and methanogenic bacteria.

The butyrate-degrading Syntrophospora bryantii degrades butyrate and a propionate-degrading strain (MPOB) degrades propionate in coculture with the hydrogen- and formate-utilizing Methanospirillum hungatii or Methanobacterium formicicum. However, the substrates are not degraded in constructed cocultures with two Methanobrevibacter arboriphilus strains which are only able to consume hydrogen. Pure cultures of the acetogenic bacteria form both hydrogen and formate during butyrate oxidation with pentenoate as electron acceptor and during propionate oxidation with fumarate as electron acceptor. Using the highest hydrogen and formate levels which can be reached by the acetogens and the lowest hydrogen and formate levels which can be maintained by the methanogens it appeared that the calculated formate diffusion rates are about 100 times higher than the calculated hydrogen diffusion rates.

Butyrates↗

Carbohydrate-binding proteins (plant/human lectins and autoantibodies from human serum) as mediators of release of lysozyme, elastase, and myeloperoxidase from human neutrophils.

Analysis of cell surface glycosylation not only provides information about cell properties such as their state of differentiation or histogenetic lineage. The carbohydrate chains also provide potentially functional binding sites to endogenous carbohydrate-binding proteins. This interaction can elicit consequent signalling processes. Because of the importance of neutrophils in the host defence system, we monitored the effect of the binding of such sugar receptors to their cell surface on the release of the enzymatic activities of lysozyme, elastase, and myeloperoxidase. Besides the mannose-binding lectin concanavalin A and the immunomodulatory alpha/beta-galactoside-binding lectin from Viscum album L., three preparations of human sugar receptors - beta-galactoside-binding lectin (M(r) 14 kDa) and two affinity-purified polyclonal IgG fractions from serum with the capacity to recognize alpha- or beta-galactosides, respectively - were used. Two animal lectins from chicken liver and intestine that bind beta-galactosides, as well as the lectin-like human serum amyloid P component, were included in order to assess the importance of slight differences in ligand recognition. Cytochalasin B-enhanced enzyme release was invariably seen with the two plant lectins and the chicken liver beta-galactoside-binding lectin, but the related intestinal lectin did not increase enzyme release. The mammalian homologue of these avian lectins triggered lysozyme secretion, and the lactoside-binding IgG fraction enhanced the amount of extracellular elastase activity slightly but significantly. Thus, the actual lectin, not the nominal specificity of sugar receptors, is crucial for elucidation of responses. Due to the highly stimulatory activity of the two plant lectins, neutrophils from patients with non-cancerous diseases and from patients with lung cancer were monitored for the extent of lectin-mediated enzyme release. Only the concanavalin A-mediated reactivity of the neutrophils was associated with the type of disease.

Adult↗

Age-related development of human anti-pig xenoantibody.

Human cytotoxic natural xenoantibodies are believed to be of the immunoglobulin M class in nature. However, a thorough understanding of the development of these natural antixenodonor xenoantibodies remains incomplete. In this study, serum samples were obtained from newborn, infant, and adult human beings. An enzyme-linked immunosorbent assay was used to determine the binding of human natural immunoglobulin M xenoantibodies to pig aortic endothelial cells and pig lymphocytes. A complement-mediated cytotoxicity assay was used to measure the cytotoxicity of newborn, infant, and adult serum to cultured pig aortic endothelial cells and pig lymphocytes. Adult human serum contained both natural immunoglobulin M and immunoglobulin G xenoantibodies to pig endothelial cells and lymphocytes, whereas newborn infant serum contained only immunoglobulin G xenoantibodies. Only adult human serum was cytotoxic to pig endothelial cells and lymphocytes. Human immunoglobulin M xenoantibodies became detectable by age 1 month. By age 2 months these natural anti-pig xenoantibodies reached serum levels equivalent to those in the human adult and resulted in similar cytotoxicity to that of adult human serum. These findings indicate that (1) natural anti-pig immunoglobulin M xenoantibodies are absent from newborn infant human serum, (2) newborn human serum is not cytotoxic to pig endothelial cells and lymphocytes despite the presence of immunoglobulin G xenoantibodies that bind to pig endothelial cells and lymphocytes, and (3) natural anti-pig immunoglobulin M xenoantibodies begin to develop as early as age 1 month and by age 2 months attain a circulating level comparable to that found in the adult.

Adult↗

Evidence for H2 and formate formation during syntrophic butyrate and propionate degradation.

Both H2 and formate were formed during butyrate oxidation by Syntrophospora bryantii with pentenoate as electron acceptor and during propionate oxidation by a mesophilic propionate oxidizing bacterium (MPOB) with fumarate as electron acceptor. H2 and formate levels were affected by the bicarbonate concentration. S bryantii and MPOB were also able to interconvert formate and H2+ HCO3-; the apparent K(M) values for formate were of 2.9 mM and 1.8 mM, respectively. The conversion of H2+ HCO3- to formate was detected only when the H2 partial pressure was above 80 kPa. This interconversion seems to be rather unimportant under conditions prevailing during syntrophic propionate and butyrate oxidation.

Journal Article↗

[Effects of Sichuan herba Epimedii on the concentration of plasma middle molecular substances and sulfhydryl group of "yang-deficiency" model animal].

The effects of tonifying the kidney and strengthening the "Yang" produced by the extracts of Chinese herbs Epimedium wushanense and E. pubescens were studied in this paper. The results showed that both two herbs could decrease the concentration of plasma middle molecular substances and increase the concentration of plasma sulfhydryl group of "Yang-deficiency" model mice, thus suggesting that the above-said tonifying and strengthening effects of Herba Epimedii may result from its effects on middle molecular substances and sulfhydryl group, so as to strengthen the body resistance and eliminate the invading pathogenic factors.

Animals↗

[An investigation on therapeutic results of extracapsular cataract extraction and intraocular lens implantation in diabetics].

44 cases (50 eyes) with diabetes and cataract were followed up for 6-30 months after extracapsular cataract extraction (ECCE) with intraocular lens implantation (IOL). No severe intra- and post-operative complications were found and 84% of eyes obtained post-operative corrected visual acuities > or = 0.5. The results suggest that the pre-operative hyperglycemia alone be not a contraindication of the operation. Pre-operatively, if the diabetes of patients with cataract can be controlled by diet or hypoglycemic agent, their glucose level in the blood remains normal and stable for a period of time and there are no other systemic and ocular complications, ECCE with IOL can be performed on such patients and the therapeutic results of the operation can be safe and reliable.

Adult↗

Affinity-purified antibodies against alpha-galactosyl residues from human serum: comparison of their binding in bovine testicular tissue with that of the Griffonia simplicifolia lectin (GSI-B4) and impact of labeling on epitope localization.

alpha-Galactosyl residues in the carbohydrate part of cellular glycoconjugates can serve as cell type-associated markers and are implicated in intercellular adhesion and biosignaling. This biological significance explains the interest to characterize probes with respective specificity as the Griffonia simplicifolia I-isolectin B4. Due to the documented occurrence of an alpha-galactoside-binding immunoglobulin G fraction in human serum we compared the extent of binding and its pattern for the lectin and the antibody using surface-immobilized extract proteins and fixed sections of bovine testicular tissue with known lectin reactivity. The antibody fractions were obtained either solely from affinity chromatography isolation on immobilized melibiose or after an additional step to deplete this fraction of galactoside-binding activities without pronounced specificity to the alpha-anomeric linkage. They yielded a rather indistinguishable reactivity in comparison to that of the lectin, when an indirect approach was used. Labeling of the antibodies with a hydrazide derivative of biotin did not affect the pattern of binding. However, significant differences were noted, when conjugation of label was targeted to amino groups via N-hydroxy-succinimide esters of biotin and digoxigenin despite performance of the modification under activity-preserving conditions. Notably, the apparent strong staining of Leydig cells and nuclei of primary spermatocytes, respectively, was not inhibitable by sugar. These differences were corroborated by a nonidentical response of the various probes in solid-phase assays with extract proteins. Thus, care should be exercised in the interpretation of histochemical data, obtained with this type of modified antibody. When these precautions are fulfilled, this immunoglobulin fraction from human serum has the potential as an alpha-galactosyl-specific histochemical tool.

Animals↗

[An experimental study on HSV-1 corneal latency by in situ nucleic acid hybridization].

An experimental study was performed on animal models for evaluation of the possibility of herpes simplex virus-1 (HSV-1) corneal latency by in situ nucleic acid hybridization. 20 normal New Zealand white (NZW) rabbits were used, and 3 x 10(6) PFU/ml of McKrae strain HSV-1 was inoculated bilaterally into the corneal stroma in 14 rabbit eyes. Of the 28 eyes, 22 developed typical herpes simplex keratitis (HSK). On the postoperative 60th day, 4 corneas with latent infection were transplanted into one unilateral eyes of each 4 non-infected NZW rabbits respectively and removed 2 weeks postoperatively. The corneal buttons were individually detected for the presence of HSV-1 antigen and nucleic acid sequences by using clonal IgG HSV-1 antibody and biotinylated HSV-1 DNA probe respectively. The results showed that the HSV-1 DNA sequences retained only within the corneal stromal layer with negative HSV-1 antigen staining. These results strongly suggest that the cornea be capable of harboring latent HSV-1.

Animals↗

[A clinical observation on intraocular lens implantation in high myopic eyes with cataract].

Extracapsular cataract extraction (ECCE) with intraocular lens (IOL) implantation was performed on 124 high myopic eyes with cataract of 101 cases whose mean period of follow-up was 16.4 months. Post-operatively, 95.97% of these eyes were relieved from blindness and 75.00%, from visual handicap. The incidence of posterior capsular opacity was 22.58% and only did 1.61% of the eyes occur retinal detachment. Intraoperatively, posterior capsular rupture occurred in 5.65% of the eyes. The results indicate that ECCE with IOL implantation performed on eyes with high myopia and cataract is effective.

Adolescent↗

[Isolation and reassociation of acetogen and methanogen in a syntrophobic coculture degrading butyrate anaerobically].

Anaerobic coculture BF2 which degraded butyrate into acetate and produced methane was isolated from granular methanogenic sludge. The coculture is associated syntrophically the Syntrophomonas subsp. saponavida strain CF2 with Methanobacterium formicicum strain MF2 and appeared to degraded C4 approximately C18 fatty acids including isobutyrate. The optimal temperature and pH for growth was 37 degrees C and 7.7 respectively. The strain CF2 was obtained in pure culture with crotonate as substrate and produces acetate and butyrate. The doubling time of strain CF2 in crotonate media was about 20 hours. Strain CF2 is Gram negative, slightly curved 0.2 approximately 0.3 x 2.0 approximately 3.0 micron with round ends, motile by lateral flagellation at the concave side, non sporeforming. With a hydrogen scavenging organism, such as Methanospillum hungatei JF1, Methanobacterium formicicum 1535, Methanobrevibacterium bryantii 1125 and Desulfovibrio sp. B11, the strain CF2 paired up and the defined coculture degraded butyrate to acetate. When the strain CF2 associated with the original accompanist, Methanobacterium formicius strain MF2, the reassociated couculture degraded butyrate to acetate and produced methane again.

Butyrates↗

A study on HSV-1 corneal potential infection by in situ nucleic acid hybridization.

PURPOSE: To evaluate the possibility of HSV-1 corneal latency by in situ nucleic acid hybridization in animal models. METHODS: 20 normal New Zealand White (NEW) rabbits were used, 14 of them were inoculated bilaterally with 3 x 10 PFU/ml of McKrae strain HSV-1 by intrastromal injection. 22/28 eyes developed typical herpes simplex keratitis (HSK) diseases. At 60 day postoperation (PI), 4 latent corneas were transplanted to one eye of 4 noninfected NZW rabbits and removed 2 weeks PI. Corneas at all time intervals of infection and two weeks after PKP were detected for presence of HSV-1 antigen and nucleic acid sequences by using clonal IgG HSV-1 antibody and biotinylated HSV-1 DNA probe individually. RESULTS: The results showed that the HSV-1 DNA sequences were retained within the corneal epithelium and anterior stromal keratocytes during acute diseases, while the corneas during latent infection and postoperation, the HSV-1 DNA sequences were retained only within the stromal layer with negative HSV-1 antigen staining. CONCLUSIONS: These results strongly suggest that the cornea may be capable of harboring latent HSV-1.

Animals↗

[A preliminary report on extracapsular cataract extraction with intraocular lens implantation in children].

Extracapsular cataract extraction (ECCE) with posterior chamber intraocular lens (IOL) implantation was performed on 94 eyes of children who were followed up for 6 approximately 30 months. IOLs were inserted in all of these eyes successfully with anterior vitrectomy used in several cases with traumatic cataract. The postoperative managements were similar to those in adults, but their complications were severer obviously than those of adults. Uveitis was the main postoperative complication which was characterized by severer anterior chamber reaction, longer duration and easier to form posterior iris synechiae and develop posterior capsular opacity. The follow-up results showed that the unilateral recovery rate from blindness is 90.1%, and 49.4% of these patients' corrected visual acuities were below 0.4. Because of the controversy on IOL implantation in children, it is the authors opinion that it should be cautious to perform IOL implantation on children and such an operation is not advocated for children under 3 years old. Children with unilateral cataract should be managed by experienced doctors, and it is very important to follow up carefully, to correct refractive error and carry out amblyopia treatment.

Adolescent↗

Abrogation of baboon natural xenoantibody to pig splenocytes by DL-penicillamine.

Natural xenoantibodies are believed to be IgM in nature and are known to play a critical role in the hyperacute rejection of distantly related xenografts. The purpose of this study was to determine whether the reducing agent DL-penicillamine could inactivate baboon natural xenoantibodies to pig splenocytes. Pooled baboon serum was treated with varying concentrations of DL-penicillamine over different lengths of time and a complement-mediated cytotoxicity assay was used to determine the reactivity of baboon natural xenoantibodies to pig splenocytes. A whole-cell ELISA assay was used to assess the binding of both IgG and IgM xenoantibodies to pig splenocytes. In addition, DL-penicillamine-treated serum was dialyzed to assess its potential clinical application. These in vitro experiments indicate that both IgM and IgG baboon natural xenoantibodies bind to pig splenocytes, but only IgM xenoantibody is cytotoxic. The binding of baboon natural IgM xenoantibody can be eliminated, and the cytotoxicity of IgM xenoantibody markedly reduced by DL-penicillamine treatment despite continued binding of IgG xenoantibody to pig splenocytes. In addition, DL-penicillamine can be dialyzed, suggesting that it may be an efficacious clinical treatment, the toxicity of which can be regulated with hemodialysis.

Animals↗

Initiation of autoimmunity to the p53 tumor suppressor protein by complexes of p53 and SV40 large T antigen.

Antinuclear antibodies (ANAs) reactive with a limited spectrum of nuclear antigens are characteristic of systemic lupus erythematosus (SLE) and other collagen vascular diseases, and are also associated with certain viral infections. The factors that initiate ANA production and determine ANA specificity are not well understood. In this study, high titer ANAs specific for the p53 tumor suppressor protein were induced in mice immunized with purified complexes of murine p53 and the Simian virus 40 large T antigen (SVT), but not in mice immunized with either protein separately. The autoantibodies to p53 in these mice were primarily of the IgG1 isotype, were not cross-reactive with SVT, and were produced at titers up to 1:25,000, without the appearance of other autoantibodies. The high levels of autoantibodies to p53 in mice immunized with p53/SVT complexes were transient, but low levels of the autoantibodies persisted. The latter may have been maintained by self antigen, since the anti-p53, but not the SVT, response in these mice could be boosted by immunizing with murine p53. Thus, once autoimmunity to p53 was established by immunizing with p53/SVT complexes, it could be maintained without a requirement for SVT. These data may be explained in at least two ways. First, altered antigen processing resulting from the formation of p53/SVT complexes might activate autoreactive T helper cells specific for cryptic epitopes of murine p53, driving anti-p53 autoantibody production. Alternatively, SVT-responsive T cells may provide intermolecular-intrastructural help to B cells specific for murine p53. In a second stage, these activated B cells might themselves process self p53, generating p53-responsive autoreactive T cells. The induction of autoantibodies during the course of an immune response directed against this naturally occurring complex of self and nonself antigens may be relevant to the generation of specific autoantibodies in viral infections, and may also have implications for understanding the pathogenesis of ANAs in SLE. In particular, our results imply that autoimmunity can be initiated by a "hit and run" mechanism in which the binding of a viral antigen to a self protein triggers an immune response that subsequently can be perpetuated by self antigen.

Animals↗

A mutation in Arabidopsis that leads to constitutive expression of systemic acquired resistance.

Systemic acquired resistance (SAR) is a nonspecific defense response in plants that is associated with an increase in the endogenous level of salicylic acid (SA) and elevated expression of pathogenesis-related (PR) genes. To identify mutants involved in the regulation of PR genes and the onset of SAR, we transformed Arabidopsis with a reporter gene containing the promoter of a beta-1,3-glucanase-encoding PR gene (BGL2) and the coding region of beta-glucuronidase (GUS). The resulting transgenic line (BGL2-GUS) was mutagenized, and the M2 progeny were scored for constitutive GUS activity. We report the characterization of one mutant, cpr1 (constitutive expressor of PR genes), that was identified in this screen and shown by RNA gel blot analysis also to have elevated expression of the endogenous PR genes BGL2, PR-1, and PR-5. Genetic analyses indicated that the phenotype conferred by cpr1 is caused by a single, recessive nuclear mutation and is suppressed in plants producing a bacterial salicylate hydroxylase, which inactivates SA. Furthermore, biochemical analysis showed that the endogenous level of SA is elevated in the mutant. Finally, the cpr1 plants were found to be resistant to the fungal pathogen Peronospora parasitica NOCO2 and the bacterial pathogen Pseudomonas syringae pv maculicola ES4326, which are virulent in wild-type BGL2-GUS plants. Because the cpr1 mutation is recessive and associated with an elevated endogenous level of SA, we propose that the CPR1 gene product acts upstream of SA as a negative regulator of SAR.

Arabidopsis↗

Anaerobic degradation of propionate by a mesophilic acetogenic bacterium in coculture and triculture with different methanogens.

A mesophilic acetogenic bacterium (MPOB) oxidized propionate to acetate and CO(2) in cocultures with the formate- and hydrogen-utilizing methanogens Methanospirillum hungatei and Methanobacterium formicicum. Propionate oxidation did not occur in cocultures with two Methanobrevibacter strains, which grew only with hydrogen. Tricultures consisting of MPOB, one of the Methanobrevibacter strains, and organisms which are able to convert formate into H(2) plus CO(2) (Desulfovibrio strain G11 or the homoacetogenic bacterium EE121) also degraded propionate. The MPOB, in the absence of methanogens, was able to couple propionate conversion to fumarate reduction. This propionate conversion was inhibited by hydrogen and by formate. Formate and hydrogen blocked the energetically unfavorable succinate oxidation to fumarate involved in propionate catabolism. Low formate and hydrogen concentrations are required for the syntrophic degradation of propionate by MPOB. In triculture with Methanospirillum hungatei and the aceticlastic Methanothrix soehngenii, propionate was degraded faster than in biculture with Methanospirillum hungatei, indicating that low acetate concentrations are favorable for propionate oxidation as well.

Journal Article↗