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Biomedical subjects

G Liang

Publications and source records attributed to G Liang.

At least 55 records · Page 3Linked to original sources
PubMed1997

Regioselective effect of zwitterionic DNA substitutions on DNA alkylation: evidence for a strong side chain orientational preference.

The incorporation of zwitterionic residues (5-substituted omega-aminoalkyl-2'-deoxypyrimidines) into DNA has been reported to bend DNA as measured by aberrant gel mobility [Strauss et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 9515-9520]. Herein we report that DNA methylation by N-methyl-N-nitrosourea at N7-guanine is regioselectively inhibited by point substitutions of the zwitterionic residues 5-(6-aminohexyl)-2'-deoxycytidine, 5-(6-aminohexyl)-2'-deoxyuridine, or 5-(3-aminopropyl)-2'-deoxyuridine. No inhibition is observed for DNA methylation by dimethyl sulfate. On the basis of inhibition patterns for methylation with the different zwitterionic substitutions and the different length tethers, the omega-aminoalkyl side chains prefer to adopt a conformation that points them toward the 3'-base. Molecular modeling grid searches, coupled with energy minimizations, and simulated annealing molecular dynamics studies indicate that unfavorable steric interactions with the 5'-base and backbone, as well as stabilizing electrostatic interactions with electronegative atoms on the 3'-side, are responsible for the observed conformational preference. No evidence for association of the cationic side chain with the phosphate backbone is observed. The observed bending of DNA induced by the tethered ammonium ions may simply arise from their localization in the major groove.

Circular Dichroism↗
PubMed1997

Identification and characterization of differentially methylated regions of genomic DNA by methylation-sensitive arbitrarily primed PCR.

We have developed a simple and reproducible fingerprinting method for screening the genome for regions of DNA that have altered patterns of DNA methylation associated with oncogenic transformation. Restriction enzymes with different sensitivities to cytosine methylation in their recognition sites were used to digest genomic DNAs from primary tumors, cell lines, and normal tissues prior to arbitrarily primed PCR amplification. Fragments that showed differential methylation were cloned and sequenced after resolving the PCR products on high-resolution polyacrylamide gels. The cloned fragments were then used as probes for Southern analysis to confirm differential methylation of these regions in colon tissues and cell lines. Forty-four DNA fragments associated with a total of five different regions of genomic DNA containing methylation sites were detected in 10 matched sets of normal and tumor colon DNAs and 7 colon cancer cell lines. A novel CpG island was also isolated that was found to be frequently hypermethylated in bladder and colon tumors. We have demonstrated that this technique is a rapid and efficient method that can be used to screen for altered methylation patterns in genomic DNA and to isolate specific sequences associated with these changes.

Blotting, Southern↗
PubMed1997

Infiltration of eosinophils into the asthmatic airways caused by interleukin 5.

Interleukin (IL)-5 is thought to play an important role in asthmatic bronchial mucosal inflammation and is a potential therapeutic target. To investigate the effect of IL-5 on the infiltration of eosinophils in airway in vivo, we compared eosinophil counts and their activation status in airways without and after the topical instillation of recombinant human IL-5. Eight subjects with mild atopic asthma underwent initial bronchoscopy during which control bronchoalveolar lavage (BAL) fluid as well as bronchial mucosa were obtained, and at the same time, normal saline and IL-5 were administered to two sublobar segments separately. The second bronchoscopy were carried out and samples from challenged sites were taken 24 h later. It was found that the total eosinophils (BMK-13+ cells) and the activated eosinophils (EG2+ cells) in bronchial mucosa, the eosinophil numbers in BAL fluid, as well as eosinophil cationic protein (ECP) in BAL fluid from saline-challenged segments were not different from those in unchallenged segments. However, a significant eosinophilia was observed in bronchial mucosa and BAL fluid from IL-5-challenged sites. Eosinophil activation, as assessed by secretion of ECP, was also increased significantly in bronchial mucosa and BAL fluid. The results strongly suggested that IL-5 is capable of inducing eosinophil infiltration into the asthmatic airways, as well as the activation of infiltrating eosinophils.

Adult↗
PubMed1997

[Experimental study on the antituberculous activity of ofloxacins].

OBJECTIVE: To compare the antituberculous activities of ofloxacins (OFLX) made by different pharmaceutical companies. METHODS: The minimal inhibitory concentrations (MIC) of OFLX were determined by two-fold dilution method. The efficacy of OFLX was compared in terms of median survival time (ST50) against M. tuberculosis infection in mice. RESULTS: In vitro the activities against M. tuberculosis (H37Rv), M. bovis (Ravenel) and M. Phlei products showed similar degree. The MICs of OFLX were 0.5-1.0 microgram/ml in modified Sauton medium and 1.0 microgram/ml in modified Lowenstin-Jensen medium. OFLX was more active than ethambutol (MIC 4-8 micrograms/ml, 2.0 micrograms/ml, respectively) against M. tuberculosis in the same media. OFLX from domestic or foreign drug companies showed similar results for treating M. tuberculosis infection in mice and all were significantly effective. The efficacy of OFLX at the dose of 4 mg per mouse was found close to that of ethambutol (EMB) at the dose of 2 mg per mouse. The efficacy of OFLX at the dose of 2 mg per mouse close to EMB at the dose of 1 mg per mouse and thiacetazone at the dose of 2 mg per mouse. CONCLUSIONS: In vitro and in vivo, the activities of OFLX from different sources against M. tuberculosis are about the same.

Animals↗
PubMed1997

[Limb salvage surgery using massive allografts in malignant bone tumors].

The advantages of segmental allografts over arthoplastics are, among others, secured biological healing, easy attachment of tendons and ligaments, and desirable long-tem results. Since 1983, the authors have performed 35 resections of malignant tumors in the extremities followed by reconstructions with implantation of segmental allografts. Five types of grafts were used for the reconstruction of the skeletal defects, intercalary hemicylinder, intercalary cylinder, partial osteoarticular, total osteoarticular, and allograft-prosthetic implant composite. All the patients underwent adjunctive chemotherapy that was considered necessary. The length of follow-up ranged from 6 months to 10 years during which time 6 patients died, 25 were disease-free, and 4 survived as tumors-carriers. Complications in 28.6% of the patients included infection, graft resorption, recurrence and nonunion. Patients undergoing intercalary grafts oblained better functional results than those having osteoarticular grafts. 4 patients had biopsies and bone scans of the grafts 2-4 years postoperation with creeping substituion being observed. Segmental allograft remains a viable approach with predictable results for reconstruction of skeletal defects after resection of malignant bone tumors.

Adolescent↗
PubMed1997

Studies on multiple forms of maltotetraose-forming amylase from Alcaligenes sp.

Zymograms of the cultural supernatant of Alcaligenes sp. showed three bands, the major one being G4A-1 and the minor two, G4A-2 and G4A-3. Based on the electrophoretic homogeneity of the purified three bands and the enzymatic activities identified by a thin layer chromatography of the soluble starch hydrolysates, all the three bands were confirmed to be maltotetraose-forming amylase but in multiple forms. Neither glycosidase nor protease activities could be detected in the culture (only very weak protease activities were observed at 48 hours after cultivation), which indicate that the two enzymes were not involved in the amylase multiple-form formation. Only the relative amount of the two minor bands (but not the multiple-form pattern) was changed when the initial pH of the medium varied from 6.5-8.5. An addition of 0.3% glucose raised the yield of G4A-2 and G4A-3.

Alcaligenes↗
PubMed1996

ATF3 gene. Genomic organization, promoter, and regulation.

ATF3 gene, which encodes a member of the activating transcription factor/cAMP responsive element binding protein (ATF/CREB) family of transcription factors, is induced by many physiological stresses. As a step toward understanding the induction mechanisms, we isolated the human ATF3 gene and analyzed its genome organization and 5'-flanking region. We found that the human ATF3 mRNA is derived from four exons distributed over 15 kilobases. Sequence analysis of the 5'-flanking region revealed a consensus TATA box and a number of transcription factor binding sites including the AP-1, ATF/CRE, NF-kappa B, E2F, and Myc/Max binding sites. As another approach to understanding the mechanisms by which the ATF3 gene is induced by stress signals, we studied the regulation of the ATF3 gene in tissue culture cells by anisomycin, an approach that has been used to study the stress responses in tissue culture cells. We showed that anisomycin at a low concentration activates the ATF3 promoter and stabilizes the ATF3 mRNA. Significantly, co-transfection of DNAs expressing ATF2 and c-Jun activates the ATF3 promoter. A possible mechanism implicating the C-Jun NH2-terminal kinase/stress-activated protein kinase (JNK/SAPK) stress-inducible signaling pathway in the induction of the ATF3 gene is discussed.

Activating Transcription Factor 3↗
PubMed1995

The use of 2-hydroperoxytetrahydrofuran as a reagent to sequence cytosine and to probe non-Watson-Crick DNA structures.

2-Hydroperoxytetrahydrofuran (THF-OOH) can be employed to sequence cytosine (C) and to probe for non-canonical DNA structures involving C. Using 32P-labeled oligomers and a DNA restriction fragment, it is demonstrated that THF-OOH has a strong preference for Cs in single-stranded (s-s) DNA regions, and in bulges, loops and mismatches. The reactivity of C is diminished below pH 6.0, but is not affected by substitution of 5-methylcytosine. To demonstrate the utility of the reagent, it is directly compared to methoxylamine and chloroacetaldehyde, two other reagents commonly used to chemically probe C residues in non-Watson-Crick DNA structures.

5-Methylcytosine↗
PubMed1995

CCAAT-box contributions to human thymidine kinase mRNA expression.

In order to examine the role of two inverted CCAAT boxes near the start of transcription of the human thymidine kinase (TK) gene, a series of constructs were prepared in which one or both CCAAT boxes were deleted or mutated. These altered promoters (1.2 kb of 5'-flanking sequence) were used to express a TK minigene containing the first two exons and introns followed by the remainder of the cDNA. RNA blots were prepared from stable cell lines of ts13 cells containing these constructs under three conditions: 1) serum deprived cells, 2) serum stimulated cells, and 3) cells that had been stimulated with serum, but were arrested in the G1 phase of the cell cycle by the temperature sensitive mutation carried by these cells. TK mRNA expression from each construct was suppressed by the temperature sensitive block to cell cycle progression. Measurement of protein expression from the various altered TK promoters indicated that both CCAAT boxes contribute to promoter strength. These experiments also suggested that the two CCAAT boxes were not equivalent and that the distal CCAAT could substitute for the proximal CCAAT, but the converse was not true.

Animals↗
PubMed1995

Protein that binds to the distal, but not to the proximal, CCAAT of the human thymidine kinase gene promoter.

Mobility shift assays were used to examine protein binding to the human TK gene CCAAT boxes. Similar protein binding patterns were observed with probes containing either the proximal or distal CCAAT. However, probes containing both CCAAT boxes in which one of the CCAAT boxes was inactivated by mutation did not demonstrate identical binding patterns. One of the complexes formed with the longer probes was only observed when the distal CCAAT was intact. This species was not formed with probes that only contained an intact proximal CCAAT, and its formation could only be competed by oligonucleotides containing the distal CCAAT motif. This observation reveals the existence of a protein that can bind to the distal, but not to the proximal, CCAAT of the human TK promoter. This protein may account for the previous observation that the two CCAAT motifs are not functionally equivalent. The protein that binds to the distal, but not to the proximal, CCAAT (DTK-CBP) was also present in two human cell lines. Significantly more DTK-CBP was present in nuclear extracts of HepG2 and WI38 cells than in TK-ts13 cells. However, this protein was not observed in three different murine cell lines and one primary culture. Its abundance in some human cell lines suggests it might modulate the expression of human TK mRNA in cells that express this protein.

3T3 Cells↗
PubMed1995

[The clinical study of local treatment of ocular malignant tumors with IL-2 LAK cells].

The authors reported the method of treatment of ocular malignant tumors at early and middle stages with local injections of self lymphokine-activated killing (LAK) cells which were induced by gamma IL-2 in vitro. Satisfactory therapeutic results were obtained. Among the 25 cases of ocular malignant tumors, 10 cases were treated only with IL-2 LAK cells, nine of them got complete regression (CR) and one got partial regression (PR); the other 15 cases were treated with operation and IL-2 LAK cells and they all obtained complete remission. In the follow-up, except the contact of 1 case with PR was lost, all the other 24 cases did not relapse after 1-5 years of observation. The above results show that the treatment of malignant tumors with local injection of self IL-2 LAK cells is a very effective immunotherapy.

Adolescent↗
PubMed1995

[Clinical behavior and cytobiological activity of desmoid tumor].

Desmoid tumor, characterized by aggressive growth and frequent recurrence without metastasis after excision, remains to be a poorly understood neoplasm. In seventeen patients with desmoid tumor of the extremities, the relationship between the cytobiological activity and prognosis of desmoid tumor was studied by reviewing clinical material, histologic examination, statistical analysis of Ag-NORs, and morphometry of tumor cell nuclei in comparison with those of fibrosarcoma and soft tissue fibroma. The results showed that the cells of desmoid tumor were highly proliferative with their biological activity bearing resemblance to fibrosarcoma. Thus we have good grounds for grouping desmoid tumors as of low grade malignancy. We believe that initial surgical resection with a wide margin is the key for good prognosis because a lower recurrence rate will result.

Adolescent↗
PubMed1995

Localization of a cycloheximide-sensitivity response element in the human thymidine kinase gene promoter.

In order to localize the segment of the human thymidine kinase (TK) gene promoter that mediates sensitivity of TK mRNA expression to the presence of cycloheximide (CX), a series of promoter truncation mutants was prepared between the 460-base pair (bp) promoter that was demonstrated previously to be sensitive to CX and the 83-bp promoter that was demonstrated previously to be insensitive to CX. TK promoters containing 370, 300, 160, or 130 bp of 5'-flanking sequence were all sensitive to inhibition by CX. Further truncation to 100 bp of 5'-flanking sequence eliminated CX sensitivity. Electrophoretic mobility shift assays using a probe containing most of this region (but omitting the SP1 binding site at the 5'-end of the 130-bp promoter) identified some complexes whose formation was sensitive to the presence of CX. Comparison of the sequences of oligonucleotides that were able to compete for formation of mobility shift complexes identified the sequence GCGGCC as a putative CX-sensitivity response element. Two such sequences are found between 83 and 130 bp 5' of the TK capsite. Mutation of the distal sequence attenuated sensitivity of TK mRNA expression to CX, while mutation of the proximal sequence had minimal effect on CX sensitivity. Thus, these data have localized a CX-sensitivity response element to a segment of TK promoter about 120 bp 5' of the capsite that includes the hexamer GCGGCC.

Base Sequence↗
PubMed1995

[Effects of a platelet-activating factor antagonist ONO-6240 on antigen induced eosinophil infiltration into the airways].

An in vivo mouse model of allergic bronchial asthma was developed to investigate the effects of a platelet activating factor antagonist ONO-6240 on eosinophil (EOS) recruitment into the mouse airway. In the present study, no EOS could be found in bronchoalveolar lavage fluid (BALF) from normal control mice. However, chronic ovalbumin challenge of sensitized mice induced significant bronchoalveolar eosinophilia (9.68 +/- 0.72 x 10(8)/L). In three groups of mice treated intraperitoneally with different doses of ONO-6240 (0.1, 1.0, 10.0 mg/kg), the medication produced 42.1% (5.60 +/- 0.97 x 10(8)/L, P < 0.01), 58.8% (3.99 +/- 0.84 x 10(8)/L, P < 0.01), and 72.6% (2.65 +/- 0.71 x 10(8)/L, P < 0.01) inhibition, respectively, of the antigen-induced influx of EOS into BALF. It was found that the prevention of EOS infiltration into airway with ONO-6240 was accompanied by decrements of both interleukin (IL)-5 and IL-2 levels. These data indicated that ONO-6240 prevented airway eosinophilia by inhibiting production of IL-5 and IL-2. Our results suggested that ONO-6240 may be of value in treating human asthmatic patients.

Animals↗