Biomedical subjects

X Yang

Publications and source records attributed to X Yang.

At least 739 records · Page 41Linked to original sources

PubMed1997

Resolution and reconstitution of succinate-ubiquinone reductase from Escherichia coli.

A modified procedure is developed for isolation of highly purified succinate-ubiquinone reductase from Escherichia coli NM256 containing a cloned sdh operon in a multicopy plasmid. Succinate-ubiquinone reductase is solubilized from the membrane by polyoxyethylene-9-lauryl ether and purified by DEAE-Sepharose CL-6B column chromatography. The isolated reductase is resolved into a reconstitutively active, two-subunit succinate dehydrogenase and a two-subunit membrane anchoring protein fraction (the SdhC-SdhD fraction) by alkaline (pH 10.2) treatment of the reductase in the presence of 1 M urea, followed by DEAE-Sepharose CL-6B column chromatography under anaerobic conditions. Isolated succinate dehydrogenase and the SdhC-SdhD fraction alone show no succinate-ubiquinone reductase activity. However, when a given amount of the SdhC-SdhD fraction is mixed with varying amounts of succinate dehydrogenase or vice versa succinate-ubiquinone reductase activity increases as the amount of succinate dehydrogenase or the SdhC-SdhD fraction added increases. Maximum reconstitution is obtained when the weight ratio of succinate dehydrogenase to the SdhC-SdhD fraction reaches 5.26. This ratio is slightly higher than the calculated value of 3.37, obtained by assuming 1 mol of succinate dehydrogenase reacts with 1 mol of SdhC and SdhD. The isolated SdhC-SdhD fraction contains 35 nmol cytochrome b556/mg protein. Unlike mitochondrial cytochrome b560, the cytochrome b556 is reducible by succinate in the isolated and complex forms. Furthermore, cytochrome b556 in the isolated SdhC-SdhD fraction has absorption properties, carbon monoxide reactivity, and EPR characteristics similar to those of cytochrome b556 in intact succinate-ubiquinone reductase, indicating that its heme environments are not affected by the presence of succinate dehydrogenase. However, the redox potential of cytochrome b556 in the SdhC-SdhD fraction (22 mV) increases slightly when complexed with succinate dehydrogenase (34 mV). No hybrid succinate-ubiquinone reductase is formed from mitochondrial QPs (the membrane-anchoring protein fraction of bovine heart mitochondrial succinate-ubiquinone reductase) and E. coli succinate dehydrogenase or vice versa. However, the cytochrome b556 in E. coli SdhC-SdhD fraction is reducible by succinate in the presence of mitochondrial succinate dehydrogenase, and the rate of cytochrome b556 reduction correlates with the reconstitutive activity of the mitochondrial succinate dehydrogenase.

Animals↗

PubMed1997

Macrophage-derived metalloelastase is responsible for the generation of angiostatin in Lewis lung carcinoma.

To determine the mechanism responsible for the in vivo production of angiostatin that inhibits growth and metastasis in Lewis lung carcinoma (3LL), we implanted 3LL variant cells into the subcutis of syngeneic C57BL/6 mice. The tumors were infiltrated by macrophages and expressed high levels of steady-state mRNA for metalloelastase (MME). Successive passages (more than three) of cultures established from the tumors resulted in complete depletion of macrophages; steady-state MME mRNA, elastinolytic activity, and production of angiostatin (in the presence of plasminogen) were correspondingly reduced. Coculture of macrophages with either 3LL cells or their conditioned media containing granulocyte-macrophage colony-stimulating factor resulted in secretion of MME and production of angiostatin by the macrophages, suggesting that angiostatin is produced by tumor-infiltrating macrophages whose MME expression is stimulated by tumor cell-derived granulocyte-macrophage colony-stimulating factor.

Angiostatins↗

PubMed1997

Proteolipid protein regulates the survival and differentiation of oligodendrocytes.

Proteolipid protein (PLP) has been postulated to play a critical role in the early differentiation of oligodendrocytes (OLs) in addition to its known role as a structural component of myelin. To identify this early function, we blocked the synthesis of PLP in glial cultures with antisense oligodeoxynucleotides that targeted the PLP initiation codon. Primary glial cultures were incubated with phosphorothioate-protected oligodeoxynucleotides (S-ODNs) for up to 11 d. PLP in OLs was reduced >90%. OLs treated with antisense S-ODNs appeared strikingly healthy as judged by (1) immunocytochemical staining for myelin glycolipids and myelin basic protein, (2) their prolonged survival compared with untreated cultures, and (3) their ability to re-establish membrane sheets after removal of the S-ODNs. Our studies show that PLP is required for elaboration and stability of the myelin membrane sheets made by most OLs, but it is not necessary for the network of processes established by OLs. More importantly, the number of OLs in the antisense-treated cultures was nearly sevenfold greater after a 10-11 d incubation with S-ODNs than in control cultures. The number of proliferating OL progenitors was not increased in the antisense-treated cultures, indicating that the increase in the number of OLs was attributable to prolonged OL survival. The tissue culture studies reveal that the absence of PLP/DM20 has the positive effect of promoting OL survival but the negative effect of preventing their full differentiation. This finding clarifies many of the paradoxical findings seen in the PLP mutants, the PLP overexpressers, and the PLP- animals.

Animals↗

PubMed1997

Characterization of human cyclooxygenase 2 gene promoter localization of a TGF-beta response element.

We characterized 2.3 kb of 5'-flanking region of the human cyclooxygenase 2 gene and analyzed its promoter activity. We localized three positive and two negative cis-acting regulatory regions and a TGF-beta response region. Basal promoter activity was located between -77 and +99 bp. Maximal promoter activity was observed from -1838 to +99 bp. A TGF-beta response region was found between -454 and -288 bp. This region included an NF-I site. Electrophoretic mobility shift assays showed that TGF-beta caused a marked change in the pattern and intensity of this NF-I like protein complex.

Animals↗

PubMed1997

CCR3 and CCR5 are co-receptors for HIV-1 infection of microglia.

Several members of the chemokine receptor family are used together with CD4 for HIV-1 entry into target cells. T cell line-tropic (T-tropic) HIV-1 viruses use the chemokine receptor CXCR4 as a co-receptor, whereas macrophage-tropic (M-tropic) primary viruses use CCR5 (refs 2-6). Individuals with defective CCR5 alleles exhibit resistance to HIV-1 infection, suggesting that CCR5 has an important role in vivo in HIV-1 replication. A subset of primary viruses can use CCR3 as well as CCR5 as a co-receptor, but the in vivo contribution of CCR3 to HIV-1 infection and pathogenesis is unknown. HIV-1 infects the central nervous system (CNS) and causes the dementia associated with AIDS. Here we report that the major target cells for HIV-1 infection in the CNS, the microglia, express both CCR3 and CCR5. The CCR3 ligand, eotaxin, and an anti-CCR3 antibody inhibited HIV-1 infection of microglia, as did MIP-1beta, which is a CCR5 ligand. Our results suggest that both CCR3 and CCR5 promote efficient infection of the CNS by HIV-1.

Adult↗

PubMed1997

Photo-irradiation improved functional preservation of the isolated rat heart.

BACKGROUND AND OBJECTIVE: Photo-irradiation causes a variety of effects in different cells and tissues. We hypothesized that photo-irradiation may improve cardiac preservation based on these observations. STUDY DESIGN/MATERIALS AND METHODS: For pre-storage treatment (Pre), the heart in an anesthetized open-chest rat was irradiated using an Argon-dye laser with a wavelength of 660 nm at a fluence of 16.8 J/cm2 or sham-operated. The heart was excised, perfused with Krebs-Henseleit Buffer, cardioplegically arrested, and stored by immersion at 0 degree C for 18 hrs. Functional recovery was evaluated by working reperfusion for 30 min. For post-storage treatment (Post), the isolated hearts were stored for 18 hrs at 0 degree C; laser irradiation at a fluence of 36 J/cm2 was administered during working reperfusion. Hearts which did not receive irradiation during reperfusion served as control. Furthermore, isolated cardiomyocytes were used to study laser effect on cellular ATP content, catalase activity, and nitric oxide (NO) release. RESULTS: Both Pre and Post groups showed significant improvement in recovery of aortic flow, cardiac output, and work compared to the corresponding control groups (P < 0.05). Combined Pre/Post laser treatment did not improve function. Investigation using isolated rat cardiomyocytes found that both end-storage ATP and end-reperfusion catalase activity in the laser-treated group were significantly higher than those in the untreated cells (P < 0.05). NO release increased by 15% in the laser group after 18 hrs of 37 degrees C incubation. CONCLUSION: Photo-irradiation improves functional recovery of the cold-stored rat heart possibly via conservation of ATP and antioxidant enzyme activity.

Adenosine Triphosphate↗

PubMed1997

Functional MRI BOLD signal coincides with electrical activity in the rat whisker barrels.

Functional MRI (fMRI) provides a noninvasive method for mapping brain functional activity based on blood oxygenation level dependent (BOLD) image contrast that is primarily due to localized increases in perfusion. Recently, Malonek and Grinvald (Science 272:551-554, 1996) suggested that during sustained functional activation, the increases in perfusion were spread over a much larger area than the localized electrical activity. In this study, it is demonstrated that the spatial distribution of the BOLD fMRI signal during sustained stimulation of rat whiskers has the same spatial pattern and dimension as that of neuronal electrical activity in the rat whisker barrels.

Animals↗

PubMed1997

Expression of aquaporins in the rat ocular tissue.

The aquaporins (AQPs) are a rapid growing family of water channel proteins found in animals, plants and microorganisms that raise plasma membrane water permeability required for efficient isosmotic fluid transport. Five homologs of aquaporins (AQP1, AQP2, AQP3, AQP4 and AQP5) have been identified from various mammalian tissues; the expression of these aquaporins in ocular tissues was studied. Semiquantitative expression levels of these aquaporins were determined in ciliary body, cornea, lens, retina, iris and choroid using RT-PCR. Expression levels of AQP1 are highest among the known aquaporins in each rat ocular tissue examined. In cornea, AQP1 is expressed approximately three-fold higher than AQP3 and 2.5 fold higher than AQP5. However, the highest intraocular expressions of AQP3 and AQP5 are in the cornea. In the iris, expression levels of AQP1 are approximately 600-700 fold higher than AQP4 and AQP5. In the ciliary body, the expression levels of AQP1 are approximately ten-fold higher than AQP4, the only other aquaporin expressed. In the lens, the major water channel is AQP1 with detectable levels of AQP4 and AQP5 that are approximately 1000-fold lower than AQP1. In choroid, AQP1 is the only water channel expressed. In retin, AQP1 is expressed approximately six-fold higher than AQP4, the only other aquaporin expressed. However, the highest ocular expression of AQP4 is in retina. AQP2 is not detected in the eye. Finally, the possible physiological roles of aquaporins in maintaining and regulating the aqueous flow, and corneal and lens transparency are discussed.

Animals↗

PubMed1997

The cure of acute Babesiasis perroncitoi in swine.

From June to August in 1988, a total of 64 pure-bred Landrace, Yorkshire (large white) and Inner Mongolian Black Swine from a farm in Huhehot were affected with a disease, 13 died. The pathogen was confirmed primarily to be Babesia perroncitoi. The drugs used for treatment were Berenil and Acaprin. The intramuscular dose for Berenil was 3 mg per kilogram weight. The subcutaneous dose for Acaprin was 0.8 mg per kilogram weight. All of which had satisfactory results. In the same season in the second year and the third year, the disease was also reported, but all sick animals were cured.

Animals↗

PubMed1997

The factors related to fungal peritonitis in patients on peritoneal dialysis.

Fungal peritonitis (FP) is a serious infectious complication of peritoneal dialysis (PD). This retrospective study was conducted in 11 cases of FP among 64 cases of patients with bacterial peritonitis (BP). Our results showed that age and sex underlying disease did not correlate significantly with the development of FP (P > 0.05), while long-term, repeated administration of antibiotics did (P < 0.01). It is suggested that the patients recently suffering from BP and being resistant to antibiotics were at great risk of suffering from FP. The key to prevent FP was to avoid BP, to use sensitive antibiotics with appropriate courses and to give nutritive treatment.

Aged↗

PubMed1997

Lipids, lipoproteins and apolipoproteins abnormalities in patients undergoing dialysis.

Twenty hemodialysis (HD) patients and 20 patients on continuous ambulatory peritoneal dialysis (CAPD) were investigated for lipids, lipoproteins and apolipoproteins abnormalities. HD patients had elevated serum triglyceride, decreased high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A-I (Apo A-I), while CAPD patients had elevated total cholesterol, triglyceride, low-density lipoprotein cholesterol (LDL-C), Apolipoprotein B (Apo B), Apo B/Apo A-I ratio, and decreased HDL-C, Apo A-I. Because of the molecular sieving effects of peritoneum, CAPD have a negative effect on these abnormalities. CAPD patients might be at greater risk of developing coronary artery disease than HD patients who are also at increased risk as compared with normals.

Aged↗

PubMed1997

Biodegradation of methyl t-butyl ether by pure bacterial cultures.

Three pure bacterial cultures degrading methyl t-butyl ether (MTBE) were isolated from activated sludge and fruit of the Gingko tree. They have been classified as belonging to the genuses Methylobacterium, Rhodococcus, and Arthrobacter. These cultures degraded 60 ppm MTBE in 1-2 weeks of incubation at 23-25 degrees C. The growth of the isolates on MTBE as sole carbon source is very slow compared with growth on nutrient-rich medium. Uniformly-labeled [14C]MTBE was used to determine 14CO2 evolution. Within 7 days of incubation, about 8% of the initial radioactivity was evolved as 14CO2. These strains also grow on t-butanol, butyl formate, isopropanol, acetone and pyruvate as carbon sources. The presence of these compounds in combination with MTBE decreased the degradation of MTBE. The cultures pregrown on pyruvate resulted in a reduction in 14CO2 evolution from [14C]MTBE. The availability of pure cultures will allow the determination of the pathway intermediates and the rate-limiting steps in the degradation of MTBE.

Arthrobacter↗

PubMed1997

Pleural signs of small peripheral pulmonary masses: pathologic correlation with chest radiographs and diagnostic value.

OBJECTIVE: The current interest in 'cost-effectiveness' of medical imagings means that chest radiography is the primary radiological examination of lung diseases. We attempted to investigate the pathological basis of different pleural signs, and determine their usefulness in differentiating small bronchogenic carcinomas from tuberculomas by using chest radiographs. MATERIALS AND METHODS: We prospectively studied 30 small solitary pulmonary masses, including 24 carcinomas and 6 tuberculomas. All 30 resected lobe specimens were investigated with preoperative radiographs, postoperative specimen radiographs, 10-15 microns thick whole-mount sections, and 5 microns thick slices for conventional histological examination. To evaluate the clinical usefulness of these pleural signs, we also retrospectively reviewed 100 chest radiographs and conventional tomographs of 1-3 cm peripheral pulmonary lesions, including 60 bronchogenic carcinomas and 40 tuberculomas. RESULTS: The results confirmed that, besides the previously known interlobular septal fibrosis with pleural retraction, the pathologic basis also included the pleural puckering and adhesion with fluid collection for the 'tail' and 'rabbit ears' signs, and pleural dimpling with fluid collection and peripheral fibrous hyperplasia for the 'triangular shadow' signs. In the retrospective group, the pleural signs with adjacent pleural thickening was significantly more frequent in tuberculomas than in carcinomas (P < 0.05, chi 2-test). CONCLUSION: Pleural signs associated with adjacent pleural thickening seems to contribute to differentiating granulomas from carcinomas. The pathological basis for this radiological finding includes visceral pleural fibrosis with the inflammatory cell infiltration, and the wide and shallow pleural dimpling filled with reactive pleural fluid.

Carcinoma, Bronchogenic↗

PubMed1997

Conformational studies of antisense DNA by PFG NMR.

Pulsed field gradient diffusion constant measurements were used to resolve the ambiguity in determining the conformational states of single-stranded DNA dodecanucleotides (d1s, d4s and d5s). For d1s and d5s, because of the spectral symmetry conventional NMR analyses cannot differentiate whether they are hairpins or homo-duplexes. However, the diffusion constants of these sequences at 300 K are 1.4 times greater than those of the comparison complementary duplexes. This result agrees well with what is expected for Dhairpin/Dduplex based on classic liquid-phase translational diffusion models and the Einstein-Stokes equation, confirming that d1s and d5s form hairpins. d4s did not show a structured spectral pattern, but its diffusion constant measurement suggests that this sequence may not be a random coil. The DNA sequences studied contain chemically modified backbone linkages and are potential antisense agents for gene regulation. The knowledge of their diffusion constants, in combination with conventional NMR analysis and other biophysical spectroscopic measurements, provides new insights into the relationships of chemical structure and conformational preference of antisense oligonucleotides and their analogs.

Animals↗

PubMed1997

Induction of p16 during immortalization by HPV 16 and 18 and not during malignant transformation.

The p16 (MTS1) tumour-suppressor gene is a cyclin-dependent kinase (cdk) inhibitor that decelerates the cell cycle by inactivating the cdks that phosphorylate the retinoblastoma tumour-suppressor gene (Rb) protein (pRb). In cervical cancers, pRb is inactivated by the HPV E7 oncoprotein or by mutations. The hypothesis of earlier reports was that the disruption of the p16/cdk-cyclin/Rb cascade is essential for malignant cervical transformation/carcinogenesis. We previously established in vitro model systems of cervical cancer representing four steps of oncogenic progression initiated by the two most common oncogenic HPVs in ectocervical and endocervical epithelial cells. This report used these systems to investigate the role of p16 in cervical cancers. A dramatic enhancement of the p16 RNA level was observed after immortalization by HPV 16 or 18. Furthermore, the p16 protein was newly observed following immortalization. However, no further changes were found for RNA or protein levels after serum selection or malignant transformation. For three cervical carcinoma cell lines, similar high levels of p16 expression were seen. Point mutations or homozygous deletions of p16 were not observed in the in vitro systems or in clinical specimens. These results suggest that the inactivation of the p16/cdk-cyclin/Rb cascade does not occur during malignant transformation but occurs during the immortalization by HPV in HPV-harbouring premalignant lesions, the in situ equivalent of immortalized cells. Also suggested is that p16 has no role in the specific malignant transformation step from immortal premalignant lesions during the carcinogenesis of HPV-initiated cervical cancers.

Adenocarcinoma↗

PubMed1997

Optosensor for cinchona alkaloids with C18 silica gel as a substrate.

A flow-through optosensor for cinchona alkaloids with C18 silica gel as a substrate is proposed. The sensor is developed in conjunction with a flow-injection analysis system and is based on the retention of the cinchona alkaloids on a C18 column and the enhancement of their fluorescence. The analytical performance characteristics of the proposed sensor for the detection and quantification of these alkaloids were as follows: the detection limits of quinine, cinchonine, quinidine and cinchonidine were 2.3, 31.6, 2.3 and 31.6 ng ml-1, respectively, with relative standard deviations of 0.9% for quinine and quinidine (20 ng ml-1, n = 7) and 1.1% for cinchonine and cinchonidine (4.0 micrograms ml-1, n =7), respectively. Most of the common species did not interfere. The recommended method has been successfully tested for determination of quinine in pharmaceutical preparations and soft drinks.

Beverages↗

PubMed1997

Enantiomeric separation and sensitive detection of propranolol, metoprolol and atenolol derivatized with a fluorogenic reagent, 4-(N-chloroformylmethyl-N-methyl)amino-7-N,N-dimethylaminosulf onyl-2,1, 3-benzoxadiazole (DBD-COCl), on cellulose chiral columns in the reversed-phase mode.

An electrophilic fluorogenic reagent, 4-(N-chloroformylmethyl-N-methyl)amino-7-N,N-dimethylaminosulfo nyl-2,1,1,3-benzoxadiazole (DBD-COCl), was used to react with three beta-blockers, propranolol, metoprolol and atenolol, to yield the derivatives which were subsequently verified to be amide compounds (1:1 adducts) derived from the reaction with the secondary amino group of the beta-blockers. DBD-COCl exhibited high reactivity for the reaction completed within 5 min under mild conditions and without any need for a catalyst. The derivatives were well separated enantiomerically on cellulose CSPs (the derivative of propranolol on a Chiralcel OD-R column and those of metoprolol and atenolol on a Chiralcel OJ-R column) in the reversed-phase mode, with the S-configurations being eluted before the R-configurations. The derivatives were stable at 4 degrees C for 1 week. The detection limits (S/N = 3) were 50 fmol for both (S)- and (R)-propranolol, 12 and 17 fmol for (S)- and (R)-metoprolol, respectively, and 15 and 20 fmol for (S)- and (R)-atenolol, respectively.

Adrenergic beta-Antagonists↗

PubMed1997

Interferon-gamma (IFN-gamma) regulates production of IL-10 and IL-12 in human herpesvirus-6 (HHV-6)-infected monocyte/macrophage lineage.

To determine whether HHV-6 infection induces expression and production of IL-10 and IL-12 in monocytes/macrophages, and to explore the influence of IFN-gamma on cytokine production in HHV-6-infected cells, expression and production of IL-10 and IL-12 were evaluated through reverse transcription-polymerase chain reaction (RT-PCR) and sandwich ELISA. HHV-6 infection induced the expression and the production of IL-10 and IL-12 in monocytes and THP-1 cells. Kinetic study showed that the expression of IL-12 mRNA decreased with accumulation of IL-10 mRNA. Expression and production of IL-12 were markedly increased when anti-human IL-10 MoAbs were added to the cultures, implying that endogenous IL-10 induced by HHV-6 inhibited IL-12 production. Addition of increasing concentrations of IFN-gamma to the cultures of HHV-6-infected cells enhanced the expression of IL-12 gene, while the accumulation of IL-10 mRNA was down-regulated. Determination of protein levels of IL-10 and IL-12 by ELISA also showed that IFN-gamma increased IL-12 and decreased IL-10 production. These results suggest that IFN-gamma regulates the production of IL-10 and IL-12 at transcriptional level mainly through inhibiting endogenous IL-10 production in HHV-6-infected monocyte/macrophage lineage.

Antibodies, Blocking↗